WO2012175991A1 - Fused pentacyclic anti - proliferative compounds - Google Patents

Fused pentacyclic anti - proliferative compounds Download PDF

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Publication number
WO2012175991A1
WO2012175991A1 PCT/GB2012/051467 GB2012051467W WO2012175991A1 WO 2012175991 A1 WO2012175991 A1 WO 2012175991A1 GB 2012051467 W GB2012051467 W GB 2012051467W WO 2012175991 A1 WO2012175991 A1 WO 2012175991A1
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Prior art keywords
ium
phenanthridin
methylchromeno
methyl
alkyl
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PCT/GB2012/051467
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French (fr)
Inventor
David Cousin
Mark Frigerio
Marc Geoffery Hummersone
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Pharminox Limited
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Priority claimed from GBGB1110727.3A external-priority patent/GB201110727D0/en
Priority claimed from GBGB1205504.2A external-priority patent/GB201205504D0/en
Application filed by Pharminox Limited filed Critical Pharminox Limited
Publication of WO2012175991A1 publication Critical patent/WO2012175991A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/06—Peri-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/06—Peri-condensed systems

Definitions

  • the present invention relates to novel compounds that function as inhibitors of the activity of the enzyme telomerase, either directly or indirectly via interaction with the telomere itself. More specifically, the present invention relates to novel compounds which are potentially useful therapeutic agents for the treatment and/or prevention of proliferative diseases, such as cancer. The present invention also relates to processes for the preparation of these compounds, to pharmaceutical compositions comprising them and to their therapeutic uses.
  • Cancer is caused by uncontrolled and unregulated cellular proliferation. Precisely what causes a cell to become malignant and proliferate in an uncontrolled and unregulated manner has been the focus of intense research over recent decades. This research has led to the identification of numerous tumour- specific or tumour-associated targets, which can then be targeted by anticancer agents.
  • telomere enzyme catalyses the synthesis of telomeres.
  • Telomeres are nucleoprotein structures at the ends of linear chromosomes which consist of DNA sequences arranged in tandemly repeated units which extend from less than 100 to several thousands of bases. With each cell division cycle, bases are lost from the ends of the chromosomes and, as these telomeres shorten in length, the cell eventually reaches a point at which apoptosis is triggered.
  • Telomerase is a ribonucleoprotein reverse transcriptase that functions to maintain the telomere length above the point at which apoptosis is triggered by utilising its internal RNA template to synthesize telomeric DNA sequences, which compensate for the loss of telomeric DNA (see Blackburn; Annu. Rev.
  • telomerase is present in stem and germ line cells of normal tissues, but is absent in most other cells.
  • telomerase activity and the associated shortened but stabilized telomeres, have been detected in the majority of tumours examined (and in over 90% of all human cancers examined).
  • the telomerase enzyme is a particularly interesting target for anti-cancer therapy.
  • the absence of telomerase in most normal cells makes this enzyme a particularly attractive target because its inhibition would be expected to cause minimal damage to normal healthy cells and tissues.
  • tumour cells have shorter telomeres and higher proliferation rates than normal replicative cell populations suggests that a therapeutic telomerase inhibitor may cause tumour cell death well before damage to regenerative tissues occurs, thereby minimising undesirable side effects.
  • telomeres and telomerase A more detailed discussion of telomeres and telomerase, and their role as anti-proliferative targets is provided in, for example, Sharma et al, Annals of Oncology, 1997, Vol. 8, pp 689-685; Urquidi et al, Ann. Med., 1998, Vol. 30, pp 419-430; Perry etal, Exp. Opin. Ther. Patents, 1998, Vol. 8, No. 12, pp 1567-1586; Autexier, Chemistry & Biology, 1999, Vol. 6, pp R299-R303; andNeidle i3 ⁇ 4a/., Anti-Cancer Drug Design, 1999, Vol. 14, pp 341-347.
  • telomere enzyme Certain compounds that target the telomerase enzyme are known.
  • WO 02/30932 describes certain N 8 , N 13 -disubstituted quino[4,3,2-KL]acridium salts that function as inhibitors of telomerase.
  • telomerase inhibitors that possess one or more advantageous pharmaceutical properties and which are potentially useful therapeutic agents for the treatment of proliferative disorders such as cancer.
  • telomerase inhibitors which exhibit good selectivity for the target as well as reduced non-target activity.
  • telomerase inhibitors that can be used as pharmaceutical agents for use in the treatment of proliferative disorders such as cancer.
  • the present invention provides a compound as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein, in admixture with a pharmaceutically acceptable diluent or carrier.
  • the present invention provides a method of inhibiting telomerase in vitro or in vivo, said method comprising contacting a cell with an effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein.
  • the present invention provides a method of inhibiting cell proliferation in vitro or in vivo, said method comprising contacting a cell with an effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein.
  • the present invention provides a method of treating a proliferative disorder in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein.
  • the present invention provides a method of treating cancer in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein.
  • the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein for use in therapy.
  • the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of a proliferative condition.
  • the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of cancer.
  • the cancer is a human cancer.
  • the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein for use in the production of a telomerase inhibitory effect.
  • the present invention provides the use of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein in the manufacture of a medicament for use in the treatment of a proliferative condition.
  • the present invention provides the use of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein in the manufacture of a medicament for use in the treatment of cancer.
  • the medicament is for use in the treatment of human cancers.
  • the present invention provides the use of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein in the manufacture of a medicament for use in the production of a telomerase inhibitory effect.
  • the present invention further provides a method of synthesising a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein.
  • the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, obtainable by, or obtained by, or directly obtained by a method of synthesis as defined herein.
  • the present invention provides novel intermediates as defined herein which are suitable for use in any one of the synthetic methods set out herein.
  • references to "treating” or “treatment” include prophylaxis as well as the alleviation of established symptoms of a condition.
  • Treating” or “treatment” of a state, disorder or condition therefore includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in a human that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition, (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical or subclinical symptom thereof, or (3) relieving or attenuating the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms.
  • a “therapeutically effective amount” means the amount of a compound that, when administered to a mammal for treating a disease, is sufficient to effect such treatment for the disease.
  • the “therapeutically effective amount” will vary depending on the compound, the disease and its severity and the age, weight, etc., of the mammal to be treated.
  • alkyl includes both straight and branched chain alkyl groups. References to individual alkyl groups such as “propyl” are specific for the straight chain version only and references to individual branched chain alkyl groups such as “isopropyl” are specific for the branched chain version only.
  • (l-6C)alkyl includes (l-4C)alkyl, (l-3C)alkyl, propyl, isopropyl and i-butyl.
  • phenyl(l-6C)alkyl includes phenyl(l-4C)alkyl, benzyl, 1-phenylethyl and 2-phenylethyl.
  • (m-nC) or "(m-nC) group” used alone or as a prefix, refers to any group having m to n carbon atoms.
  • alkylene is an alkyl, alkenyl, or alkynyl group that is positioned between and serves to connect two other chemical groups.
  • (1- 6C)alkylene means a linear saturated divalent hydrocarbon radical of one to six carbon atoms or a branched saturated divalent hydrocarbon radical of three to six carbon atoms, for example, methylene, ethylene, propylene, 2-methylpropylene, pentylene, and the like.
  • (2-6C)alkenylene means a linear divalent hydrocarbon radical of two to six carbon atoms or a branched divalent hydrocarbon radical of three to six carbon atoms, containing at least one double bond, for example, as in ethenylene, 2,4-pentadienylene, and the like.
  • (2-6C)alkynylene means a linear divalent hydrocarbon radical of two to six carbon atoms or a branched divalent hydrocarbon radical of three to six carbon atoms, containing at least one triple bond, for example, as in ethynylene, propynylene, and butynylene and the like.
  • (3-8C)cycloalkyl means a hydrocarbon ring containing from 3 to 8 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or bicyclo[2.2.1]heptyl.
  • (3-8C)cycloalkenyl means a hydrocarbon ring containing at least one double bond, for example, cyclobutenyl, cyclopentenyl, cyclohexenyl or cycloheptenyl, such as 3-cyclohexen-l- yl, or cyclooctenyl.
  • (3-8C)cycloalkyl-(l-6C)alkylene means a (3-8C)cycloalkyl group covalently attached to a (l-6C)alkylene group, both of which are defined herein.
  • halo refers to fluoro, chloro, bromo and iodo.
  • heterocyclyl means a non-aromatic saturated or partially saturated monocyclic, fused, bridged, or spiro bicyclic heterocyclic ring system(s).
  • heterocyclyl includes both monovalent species and divalent species.
  • Monocyclic heterocyclic rings contain from about 3 to 12 (suitably from 3 to 7) ring atoms, with from 1 to 5 (suitably 1, 2 or 3) heteroatoms selected from nitrogen, oxygen or sulfur in the ring.
  • Bicyclic heterocycles contain from 7 to 17 member atoms, suitably 7 to 12 member atoms, in the ring.
  • Bicyclic heterocyclic(s) rings may be fused, spiro, or bridged ring systems.
  • heterocyclic groups include cyclic ethers such as oxiranyl, oxetanyl, tetrahydrofuranyl, dioxanyl, and substituted cyclic ethers.
  • Heterocycles containing nitrogen include, for example, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, tetrahydrotriazinyl, tetrahydropyrazolyl, and the like.
  • Typical sulfur containing heterocycles include tetrahydrothienyl, dihydro-l ,3-dithiol, tetrahydro- 2H-thiopyran, and hexahydrothiepine.
  • Other heterocycles include dihydro-oxathiolyl, tetrahydro-oxazolyl, tetrahydro-oxadiazolyl, tetrahydrodioxazolyl, tetrahydro-oxathiazolyl, hexahydrotriazinyl, tetrahydro-oxazinyl, morpholinyl, thiomorpholinyl, tetrahydropyrimidinyl, dioxolinyl, octahydrobenzofuranyl, octahydrobenzimidazolyl, and octahydrobenzothiazolyl.
  • heterocycles containing sulfur the oxidized sulfur heterocycles containing SO or S0 2 groups are also included.
  • examples include the sulfoxide and sulfone forms of tetrahydrothienyl and fhiomorpholinyl such as tetrahydrothiene 1,1 -dioxide and thiomorpholinyl 1,1 -dioxide.
  • heterocyclyl groups are saturated monocyclic 3 to 7 membered heterocyclyls containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen or sulfur, for example azetidinyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, tetrahydrothienyl, tetrahydrothienyl 1,1 -dioxide, thiomorpholinyl, thiomorpholinyl 1,1 -dioxide, piperidinyl, homopiperidinyl, piperazinyl or homopiperazinyl.
  • any heterocycle may be linked to another group via any suitable atom, such as via a carbon or nitrogen atom.
  • reference herein to piperidino or morpholino refers to a piperidin-l-yl or morpholin-4-yl ring that is linked via the ring nitrogen.
  • bridged ring systems is meant ring systems in which two rings share more than two atoms, see for example Advanced Organic Chemistry, by Jerry March, 4 th Edition, Wiley Interscience, pages 131-133, 1992.
  • bridged heterocyclyl ring systems include, aza- bicyclo[2.2.1]heptane, 2-oxa-5-azabicyclo[2.2.1]heptane, aza-bicyclo[2.2.2]octane, aza- bicyclo [3.2.1] octane and quinuclidine.
  • Heterocyclyl(l-6C)alkyl means a heterocyclyl group covalently attached to a (1- 6C)alkylene group, both of which are defined herein.
  • heteroaryl or “heteroaromatic” means an aromatic mono-, bi-, or polycyclic ring incorporating one or more (for example 1-4, particularly 1, 2 or 3) heteroatoms selected from nitrogen, oxygen or sulfur.
  • heteroaryl includes both monovalent species and divalent species. Examples of heteroaryl groups are monocyclic and bicyclic groups containing from five to twelve ring members, and more usually from five to ten ring members.
  • the heteroaryl group can be, for example, a 5- or 6-membered monocyclic ring or a 9- or 10- membered bicyclic ring, for example a bicyclic structure formed from fused five and six membered rings or two fused six membered rings.
  • Each ring may contain up to about four heteroatoms typically selected from nitrogen, sulfur and oxygen.
  • the heteroaryl ring will contain up to 3 heteroatoms, more usually up to 2, for example a single heteroatom.
  • the heteroaryl ring contains at least one ring nitrogen atom.
  • the nitrogen atoms in the heteroaryl rings can be basic, as in the case of an imidazole or pyridine, or essentially non- basic as in the case of an indole or pyrrole nitrogen. In general the number of basic nitrogen atoms present in the heteroaryl group, including any amino group substituents of the ring, will be less than five.
  • heteroaryl examples include furyl, pyrrolyl, thienyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,3,5-triazenyl, benzofuranyl, indolyl, isoindolyl, benzothienyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, benzothiazolyl, indazolyl, purinyl, benzofurazanyl, quinolyl, isoquinolyl, quinazolinyl, quinoxalinyl, cinnolinyl, pteridinyl, naphthyridin
  • Heteroaryl also covers partially aromatic bi- or polycyclic ring systems wherein at least one ring is an aromatic ring and one or more of the other ring(s) is a non-aromatic, saturated or partially saturated ring, provided at least one ring contains one or more heteroatoms selected from nitrogen, oxygen or sulfur.
  • partially aromatic heteroaryl groups include for example, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 2-oxo- 1,2,3,4-tetrahydroquinolinyl, dihydrobenzthienyl, dihydrobenzfuranyl, 2,3-dihydro- benzo[l,4]dioxinyl, benzo[l,3]dioxolyl, 2,2-dioxo-l,3-dihydro-2-benzothienyl, 4,5,6,7- tetrahydrobenzofuranyl, indolinyl, l,2,3,4-tetrahydro-l,8-naphthyridinyl, l,2,3,4-tetrahydropyrido[2,3- ?]pyrazinyl and 3,4-dihydro-2H-pyrido[3,2-Z?] [l,4]oxazinyl
  • heteroaryl groups examples include but are not limited to pyrrolyl, furanyl, thienyl, imidazolyl, furazanyl, oxazolyl, oxadiazolyl, oxatriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl and tetrazolyl groups.
  • heteroaryl groups examples include but are not limited to pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl and triazinyl.
  • a bicyclic heteroaryl group may be, for example, a group selected from:
  • bicyclic heteroaryl groups containing a six membered ring fused to a five membered ring include but are not limited to benzofuranyl, benzothiophenyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl,
  • bicyclic heteroaryl groups containing two fused six membered rings include, but are not limited to, quinolinyl, isoquinolinyl, chromanyl, thiochromanyl, chromenyl, isochromenyl, chromanyl, isochromanyl, benzodioxanyl, quinolizinyl, benzoxazinyl, benzodiazinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl and pteridinyl groups.
  • Heteroaryl(l-6C)alkyl means a heteroaryl group covalently attached to a (1- 6C)alkylene group, both of which are defined herein.
  • heteroaralkyl groups include pyridin-3-ylmethyl, 3-(benzofuran-2-yl)propyl, and the like.
  • aryl means a cyclic or polycyclic aromatic ring having from 5 to 12 carbon atoms.
  • aryl includes both monovalent species and divalent species. Examples of aryl groups include, but are not limited to, phenyl, biphenyl, naphthyl and the like. In particular embodiment, an aryl is phenyl.
  • aryl(l-6C)alkyl means an aryl group covalently attached to a (l-6C)alkylene group, both of which are defined herein.
  • aryl-(l-6C)alkyl groups include benzyl, phenylethyl, and the like
  • heterocyclyl(m-nC)alkyl comprises (m-nC)alkyl substituted by
  • heterocyclyl refers to either groups, structures, or molecules that are substituted and those that are not substituted.
  • X is O, S, SO or S0 2 ;
  • Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl, (2-6C)alkynyl, or Q is group of the formula:
  • L 1 is absent or (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene, each of which is optionally substituted with one or more (l-4C)alkyl groups;
  • Q 1 is selected from -OR 9 , -NR9R1 0 , -S(0) p R 9 (wherein p is 0, 1 or 2), -C(0)R 9 , -C(0)OR 9 , -OC(0)R 9 , -C(O)NR 9 R 10 , -N(R 10 )C(O)R 9 , -N(R 10 )CON(R 10 )R 9 -, -S0 2 N(R 9 )-, -N(R 9 )S0 2 -, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • R is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (1- 4C)alkoxy;
  • Rio is selected from hydrogen or (l-6C)alkyl
  • Ri, R2, R3, R4, R5, R 6 , R7 Re, R21, R22 and R23 are each independently selected from hydrogen, halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,
  • L 2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
  • L 3 is absent or is selected from O, S, SO, S0 2 , OS(0) 2 , N(Ri 3 ), C(O), CH(ORi 3 ),
  • R 13 and R J4 are each independently selected from hydrogen or (l-4C)alkyl
  • R20 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R 2 o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl (l-4C)alkoxy, or a group of the formula:
  • L 4 is absent or is selected from O, S, SO, S0 2 , N(R 25 ), C(O), CH(OR 25 ), C(0)0, OC(O), C(0)N(R 25 ), N(R 25 )C(0), N(R 25 )C(0)N(R 2 6), S(0) 2 N(R 25 ), or N(R25)S0 2 , wherein R 2 5 and R 2 6 are each independently selected from hydrogen or (l-4C)alkyl; and
  • R 24 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R 2 4 is optionally further substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl or (l-4C)alkoxy;
  • any nitrogen atoms present in any of the groups R l5 R 2 , R 3 , R4, R5, R6, R7 Rs, R21, R22 and R2 3 are optionally in the form of an N-oxide;
  • X is O, S, SO or S0 2 ;
  • Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl or (2-6C)alkynyl, or Q is a group of the formula:
  • L 1 is absent or (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene, each of which is optionally substituted with one or more (l-4C)alkyl groups;
  • Q 1 is selected from -OR9, -NR9R1 0 , -S(0) p R 9 (wherein p is 0, 1 or 2), -C(0)R 9 , -C(0)OR 9 , -OC(0)R 9 , -C(0)NR 9 Rio, -N(Ri 0 )C(O)R 9 , -N(R 10 )CON(R 10 )R 9 -, -S0 2 N(R 9 )-, -N(R 9 )S0 2 -, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3-8C)cyclo
  • R 9 is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (1- 4C)alkoxy;
  • Rio is selected from hydrogen or (l-6C)alkyl
  • Ri, R 2 , R3, R4, R5, R 6 , R7 and Rg are each independently selected from hydrogen, halogeno, trifluoromefhyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,
  • L 2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
  • L 3 is absent or is selected from O, S, SO, S0 2 , N(Ri 3 ), C(O), CH(ORi 3 ), C(0)0, OC(O), C(0)N(R 13 ), N(Ri 3 )C(0), N(R 13 )C(0)N(R 14 ), S(0) 2 N(R 13 ), or
  • Ri 3 and R14 are each independently selected from hydrogen or (l-4C)alkyl
  • R 20 is (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3-6C)cycloalkyl-(l-
  • R 20 is optionally further substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl or (1- 4C)alkoxy;
  • novel compounds of the invention include, for example, compounds of the formula I, or pharmaceutically acceptable salts or solvates thereof, wherein, unless otherwise stated, each of X, Q, R 1; R2, R 3 , R4, R5, R 6 , R7, Rs, R21, R22 and R23 has any of the meanings defined hereinbefore or in any of paragraphs (1) to (41) hereinafter: -
  • X is O, S or S0 2 ;
  • Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl or (2-6C)alkynyl, or Q is a group of the formula: wherein:
  • L 1 is (l-6C)alkylene which is optionally substituted with one or more (l-4C)alkyl groups;
  • Q 1 is selected from -OR 9 , -NR 9 R 10 , -S(0) p R 9 (wherein p is 0, 1 or 2), -C(0)R 9 ,
  • -C(0)OR 9 -OC(0)R 9 , -C(0)NR 9 Rio, -N(R 10 )C(O)R 9 , -N(Ri 0 )CON(Ri 0 )R 9 -, -S0 2 N(R 9 )-, -N(R 9 )S0 2 -, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3- 8C)cycloalkyl, aryl, heterocyclyl or heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • R 9 is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • Rio is selected from hydrogen or (l-6C)alkyl
  • Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl or (2-6C)alkynyl, or Q is a group of the formula:
  • L 1 is (l-4C)alkylene which is optionally substituted with one or more (l-2C)alkyl groups;
  • Q 1 is selected from -OR 9 , -NR 9 Rio, -S(0) p R 9 (wherein p is 0, 1 or 2), -C(0)R 9 ,
  • -C(0)OR 9 , -OC(0)R 9 , -C(O)NR 9 R 10 , -N(R 10 )C(O)R 9 , -N(R 10 )CON(R 10 )R 9 -, -S0 2 N(R 9 )-, -N(R 9 )S0 2 -, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3- 6C)cycloalkyl, aryl, heterocyclyl or heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • R 9 is selected from hydrogen, (l-4C)alkyl, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-4C)alkyl, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • Rio is selected from hydrogen or (l-2C)alkyl
  • Q is selected from 0 ⁇ , (l-6C)alkyl, (l-6C)alkoxy, or Q is a group of the formula: wherein:
  • L 1 is (l-2C)alkylene
  • Q 1 is selected from -OR 9 , -NR 9 Ri 0 , -S(0) p R 9 (wherein p is 0, 1 or 2), -C(0)R 9 ,
  • -C(0)OR 9 -OC(0)R 9 , -C(0)NR 9 Rio, -N(Ri 0 )C(O)R 9 , -N(Ri 0 )CON(Ri 0 )R 9 -, -S0 2 N(R 9 )-, -N(R 9 )S0 2 -, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3- 6C)cycloalkyl, aryl, heterocyclyl or heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • R9 is selected from hydrogen or (l-4C)alkyl
  • L and Q have any one of the definitions set out herein;
  • Q is selected from O , (l-6C)alkyl, or (l-6C)alkoxy;
  • Q is selected from O , (l-4C)alkyl, or (l-4C)alkoxy;
  • Q is selected from O , (l-2C)alkyl, (l-2C)alkoxy;
  • Q is O ⁇ or (l-6C)alkyl.
  • Q is O " ,or (l-2C)alkyl.
  • Q is O , or methyl.
  • Ri, R2, R3, R4, R5, R 6 , R7 and Rs are each independently selected from hydrogen,
  • halogeno trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, or a group of the formula:
  • L 2 is absent or (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
  • L 3 is absent or is selected from O, S, SO, S0 2 , N(R i3 ), C(O), CH(ORi 3 ), C(0)0,
  • Ri 3 and R14 are each independently selected from hydrogen or (l-2C)alkyl
  • R 20 is (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3-6C)cycloalkyl-(l- 6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l- 6C)alkyl and wherein R 20 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • Ri, R2, R3, R4, R5, R 6 , 7 and R 8 are each independently selected from hydrogen, halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, or a group of the formula:
  • L 2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
  • L 3 is absent or is selected from O, S, SO, S0 2 , N(Ri 3 ), C(O), CH(ORi 3 ), C(0)N(Ri 3 ), N(Ri 3 )C(0), S(0) 2 N(Ri 3 ), or N(Ri 3 )S0 2 , wherein R i3 and R i4 are each independently selected from hydrogen or (l-2C)alkyl; and
  • R 20 is (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3-6C)cycloalkyl-(l- 6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l- 6C)alkyl and wherein R2 0 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
  • R 2 i, R22 and R 23 are hydrogen
  • Ri, R2, R 3 , R4, R5, R 6 , R7 and Rg are each independently selected from hydrogen,
  • L 3 is absent or is selected from O, S, SO, S0 2 , N(R i3 ), C(O), CH(ORi 3 ), C(0)N(Ri 3 ), N(Ri 3 )C(0), N(R 13 )C(0)N(R 14 ), S(0) 2 N(Ri 3 ), or N(Ri 3 )S0 2 , wherein Ri 3 and R14 are each independently selected from hydrogen or (1- 2C)alkyl; and
  • R2 0 is (l-6C)alkyl or (3-6C)cycloalkyl, and wherein R 2 o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-2C)alkoxy;
  • R 2 i, R 22 and R 23 are hydrogen
  • Ri, R 2 , R 3 , R4, R5, R 6 , R7 and R 8 are each independently selected from hydrogen
  • L 3 is absent or is selected from O, S, SO, S0 2 , N(Ri 3 ), C(O), CH(ORi 3 ), C(0)N(Ri 3 ), N(Ri 3 )C(0), S(0) 2 N(Ri 3 ), or N(Ri 3 )S0 2 , wherein R i3 and R i4 are each independently selected from hydrogen or (l-2C)alkyl; and
  • R2 0 is (l-4C)alkyl which is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-2C)alkoxy;
  • R21, R22 and R 23 are hydrogen
  • Ri, R2, R 3 , R4, R5, R 6 , R7 Re, R21, R22 and R2 3 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
  • L 2 is absent or a (l-6C)alkylene or (2-6C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(Ri 3 ), C(0)0, C(0)N(Ri 3 ),
  • R i3 is selected from hydrogen or (l-4C)alkyl
  • R2 0 is hydrogen, (l-6C)alkyl, heteroaryl, or heterocyclyl and wherein R2 0 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-4C)alkoxy, or a group of the formula:
  • L 4 is N(R 25 ), wherein R25 is independently selected from hydrogen or (1- 4C)alkyl;
  • R 24 is (l-6C)alkyl
  • any nitrogen atoms present in any of the groups Ri, R2, R 3 , R 4 , R5, R6, R ? ,Rs R21, R22 and R2 3 are optionally in the form of a N-oxide;
  • Ri, R2, R 3 , R4, R5, R 6 , 7 s, R21, R22 and R2 3 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
  • L 2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(Ri 3 ), C(0)0, C(0)N(Ri 3 ),
  • Ri 3 is selected from hydrogen or (l-2C)alkyl
  • R2 0 is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R 2 o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy, or a group of the formula:
  • L is N(R 25 ), wherein R 25 is (l-2C)alkyl
  • R 24 is (l-2C)alkyl
  • any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7, Re, R21, R22 and R2 3 are optionally in the form of a N-oxide;
  • Ri, R2, R 3 , R4, R5, R 6 , R7 Rs, R21, R22 and R 2 3 are each independently selected from hydrogen, fluoro, hydroxy, or a group of the formula:
  • L 2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(R 13 ), C(0)0, C(0)N(Ri 3 ), N(Ri 3 )C(0), wherein R1 3 is selected from hydrogen or (l-2C)alkyl; and R20 is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R20 is optionally further substituted by one or more substituents independently selected from fluoro, hydroxy, (l-2C)alkoxy, or a group of the formula:
  • L 4 is N(R 25 ), wherein R 25 is (l-2C)alkyl
  • R 24 is (l-2C)alkyl
  • any nitrogen atoms present in any of the groups Ri, R2, R 3 , R4, R5, R6, R7, Re, R21, R22 and R2 3 are optionally in the form of a N-oxide;
  • Ri, R 2 , R 3 , R4, R5, R 6 , R7 R8, R21, R22 and R2 3 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
  • L 2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(Ri 3 ), C(0)0, N(Ri 3 )C(0), wherein Ri 3 is selected from hydrogen or (l-2C)alkyl; and R2 0 is (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R 2 o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy, or a group of the formula:
  • L is N(R 25 ), wherein R 25 is (l-2C)alkyl
  • R 24 is (l-2C)alkyl
  • any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7, Re, R21, R22 and R2 3 are optionally in the form of a N-oxide;
  • Ri, R2, R3, R4, R5, R 6 , 7 s, R21, R22 and R 2 3 are each independently selected from
  • L 2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(Ri 3 ), C(0)0, N(Ri 3 )C(0), wherein Ri 3 is selected from hydrogen or (l-2C)alkyl;
  • R20 is (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R20 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy,
  • Rg, R21, R22 and R2 3 are optionally in the form of a N-oxide
  • Ri, R2, R3, R4, R5, R 6 , 7 s, R21, R22 and R 2 3 are each independently selected from hydrogen, halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,
  • L 2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
  • L 3 is absent or is selected from O, S, SO, S0 2 , OS(0) 2 , N(R i3 ), C(O), CH(ORi 3 ), C(0)0, OC(O), C(0)N(R 13 ), N(R 13 )C(0), N(R 13 )C(0)N(R 14 ), S(0) 2 N(R 13 ), or
  • R i3 and R J4 are each independently selected from hydrogen or (l-4C)alkyl
  • R 20 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R 2 o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl (l-4C)alkoxy
  • any nitrogen atoms present in any of the groups Ri, R 2 , R 3 , R4, R5, R5, R7, Rg, R21, R 22 and R 2 3 are optionally in the form of a N-oxide;
  • Ri, R 2 , R 3 , R4, R5, R 6 , 7 s, R 2 i, 22 and R 23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
  • L 2 is absent or a (l-6C)alkylene, or (2-6C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(Ri 3 ), C(0)0, C(0)N(Ri 3 ),
  • Ri 3 is selected from hydrogen or (l-4C)alkyl
  • R2 0 is hydrogen, (l-6C)alkyl, heteroaryl, or heterocyclyl and wherein R 2 o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-4C)alkoxy
  • Rg, R 2 i, R 22 and R 23 are optionally in the form of a N-oxide
  • Ri, R 2 , R 3 , R4, R5, R 6 , 7 s, 2 i, R 22 and R 23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
  • L 2 is absent or a (l-3C)alkylene, or (2-3C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(Ri 3 ), C(0)0, C(0)N(Ri 3 ),
  • R1 3 is selected from hydrogen or (l-2C)alkyl
  • R 2 o is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R 2 o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy;
  • Rg, R21, R22 and R 2 3 are optionally in the form of a N-oxide
  • Ri, R2, R 3 , R4, R5, R 6 , 7 s, 2 i > R 22 and R 23 are each independently selected from hydrogen, fluoro, hydroxy, or a group of the formula:
  • L 2 is absent or a (l-3C)alkylene, or (2-3C)alkynylene;
  • L 3 is absent or is selected from O, OS(0) 2 , N(Ri 3 ), C(0)0, C(0)N(Ri 3 ),
  • R2 0 is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R2 0 is optionally further substituted by one or more substituents independently selected from oxo, fluoro, hydroxy, or methoxy;
  • any nitrogen atoms present in any of the groups R l5 R 2 , R3, R4, R5, R , R7 Rs, R21, R22 and R23 are optionally in the form of a N-oxide;
  • Ri and R2 are each independently a substituent group as defined hereinbefore (in
  • R3, R4, R5, R 6 , R7, Re, R21, R22 and R2 3 are each independently selected from hydrogen, fluoro, CF 3 , OCF 3 or (l-2C)alkoxy.
  • one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and R 3 , R4, R5, R5, R 7 R 8 , R 2 i, R22 and R 2 3 are each independently selected from hydrogen, fluoro, CF 3 , OCF 3 or (l-2C)alkoxy.
  • Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto four of R 3 , R4, R5, R 6 , R 7 R 8 , R 2 i, R22 and R 2 3 are each independently selected from fluoro, CF 3 , OCF 3 or (l-2C)alkoxy and the others are hydrogen.
  • Ri and R 2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto three of R3, R4, R5, R6, R 7 Rs, R21, R22 and R2 3 are each independently selected from fluoro, CF 3 , OCF 3 or (l-2C)alkoxy and the others are hydrogen.
  • Ri and R 2 are each independently a substituent group as defined hereinbefore (in
  • R 3 , R4, R5, R6, R7 Re, R 21 , R 22 and R 2 3 are each independently selected from hydrogen, fluoro, CF 3 or OCF 3 .
  • Ri and R 2 are a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and R3, R4, R5, R6, R 7 Rs, R21, R22 and R2 3 are each independently selected from hydrogen, fluoro, CF 3 or OCF 3 .
  • one of Ri and R 2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto four of R 3 , R4, R5, R6, R7 R 8 , R 2 i, R22 and R2 3 are each independently selected from fluoro, CF 3 or OCF 3 and the others are hydrogen.
  • Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto three of R 3 , R 4 , R5, R 6 , R 7 R 8 , R 2 i, R22 and R 2 3 are each independently selected from fluoro, CF 3 or OCF 3 and the others are hydrogen.
  • Ri and R2 are each independently a substituent group as defined hereinbefore (in
  • R 3 , R4, R5, R 6 , R 7 Rs, R21, R22 and R2 3 are each independently selected from hydrogen, fluoro, CF 3 or OCF 3 .
  • Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and R 3 , R4, R5, R5, R 7 Rs, R21, R22 and R 2 3 are each independently selected from hydrogen or fluoro.
  • one of Ri and R 2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto four of R 3 , R4, R5, R 6 , R 7 R 8 , R 2i , R 22 and R 23 are fluoro and the others are hydrogen.
  • one of Ri and R 2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto three of R 3 , R 4 , R5, R 6 , R 7 Rs, R21, R22 and R2 3 are fluoro and the others are hydrogen.
  • X is as defined in any one of paragraphs 1 to 4 above.
  • X is O or S.
  • Q is as defined in any one of paragraphs 5 to 16 above.
  • Q is O " or methyl.
  • Ri, R 2 , R3, R 4 , R5, R 6 , R7 Rs, R 21 , R 22 and R 2 3 have any one of the definitions set out in paragraphs 17 to 41 above.
  • R l5 R 2 , R 3 , R4, R5, R6, R 7 R 8 , R 2 i, R 22 and R 23 have any one of the definitions set out in paragraphs 29 to 41 above.
  • Ri, R 2 , R 3 , R ⁇ , R5, R 6 , R 7 Rs, R 2 i, R 22 and R2 3 have any one of the definitions set out in paragraphs 30 to 41 above.
  • up to four of R l 5 R 2 , R3, R4, R5, R 6 , R7, Re, R21, R22 and R2 3 are a substituent group other than hydrogen.
  • Ri, R2, R3, R 4 , R5, R6, R7, Re, R21, R22 and R2 3 are a substituent group other than hydrogen.
  • one or two of Ri, R2, R3, R 4 , R5, R 6 , R7, Rg, R21, R22 and R 2 3 is/are a substituent group other than hydrogen.
  • R l 5 R 2 , R3, R4, R5, R6, R7, Rs, 21 5 R22 and R23 is a substituent group other than hydrogen.
  • Rg, R21, R22 and R2 3 are hydrogen, i.e. the compounds have the structural formula II shown below:
  • X, Q, Ri, R2, R3, R4, R5, R6 and R7 have any one of the meanings defined hereinbefore, or a pharmaceutically acceptable salt or solvate thereof.
  • one of Ri or R 2 is hydrogen.
  • X is as defined herein before (and is particularly as described in any one of paragarphs (1) to (4) above);
  • Q is O " or (l-6C)alkyl (and is particularly O " or methyl);
  • Ri, R 2 and R 3 each independently have any one of the definitions set out hereinbefore (and in particular are as defined in any one of paragraphs (17) to (41) above);
  • R 4 , R5, R 6 , R 7 , Re, R21, R22 and R23 each have any one of the definitions set out hereinbefore;
  • X is O, S or S0 2 ;
  • R 2 , R3, R4, R5, R6, R7, Rs, R21, R22 and R23 each have any one of the definitions set out in paragraphs (30) to (41) above;
  • the various functional groups and substituents making up the compounds of the invention are typically chosen such that the molecular weight of the compound does not exceed 1000. More usually, the molecular weight of the compound will be less than 750, for example less than 700, or less than 650, or less than 600, or less than 550. More preferably, the molecular weight is less than 525 and, for example, is 500 or less.
  • Particular compounds of the invention include any one of the following:
  • Suitable pharmaceutically acceptable anions include, for example,
  • the compounds of the invention may be present in any pharmaceutically acceptable salt form.
  • the compounds of the invention are positively charged, they are suitably associated with one or more pharmaceutically acceptable anions.
  • the compound of the invention will suitably possess a +1 charge, although other charged substituent groups could be present.
  • the associated anion may carry a -1 charge, in which case the molar ratio of anionrcation is 1:1, or , alternatively, the anion may carry a -2 or -3 charge, in which case the molar ratio of anion ation is will be 1:2 or 1:3, respectively.
  • the anion is independently derived from one or more of the following acids: hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, phosphorous acetic, propionic, succinic, gycolic, stearic, lactic, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetyoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, trifluoromethanesulfonic, ethanesulfonic, ethane disulfonic, oxalic, isethionic, and valeric.
  • stereoisomers that are not mirror images of one another are termed “diastereomers” and those that are non-superimposable mirror images of each other are termed “enantiomers”.
  • enantiomers When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible.
  • An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (-)-isomers respectively).
  • a chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a "racemic mixture".
  • the compounds of this invention may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof.
  • the methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see discussion in Chapter 4 of "Advanced Organic Chemistry", 4th edition J. March, John Wiley and Sons, New York, 2001), for example by synthesis from optically active starting materials or by resolution of a racemic form.
  • Some of the compounds of the invention may have geometric isomeric centres (E- and Z- isomers). It is to be understood that the present invention encompasses all optical, diastereoisomers and geometric isomers and mixtures thereof that possess telomerase inhibitory activity.
  • the present invention also encompasses compounds of the invention as defined herein which comprise one or more isotopic substitutions.
  • H may be in any isotopic form, including 3 ⁇ 4, 2 H(D), and 3 H (T);
  • C may be in any isotopic form, including 12 C, 13 C, and 14 C;
  • O may be in any isotopic form, including 16 0 and 18 0; and the like.
  • tautomeric forms include keto-, enol-, and enolate-forms, as in, for example, the following tautomeric pairs: keto/enol (illustrated below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime, thioketone/enethiol, and nitro/aci-nitro.
  • N-oxides Compounds of the invention containing an amine function may also form N-oxides.
  • a reference herein to a compound of the invention that contains an amine function also includes the N-oxide.
  • one or more than one nitrogen atom may be oxidised to form an N-oxide.
  • Particular examples of N-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle.
  • N-Oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a per-acid (e.g.
  • N-oxides can be made by the procedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514) in which the amine compound is reacted with m-chloroperoxybenzoic acid (MCPBA), for example, in an inert solvent such as dichloromethane.
  • MCPBA m-chloroperoxybenzoic acid
  • the compounds of the invention may be administered in the form of a pro-drug which is broken down in the human or animal body to release a compound of the invention.
  • a pro-drug may be used to alter the physical properties and/or the pharmacokinetic properties of a compound of the invention.
  • a pro-drug can be formed when the compound of the invention contains a suitable group or substituent to which a property-modifying group can be attached.
  • Examples of pro-drugs include in vivo cleavable ester derivatives that may be formed at a carboxy group or a hydroxy group in a compound of the invention and in-vivo cleavable amide derivatives that may be formed at a carboxy group or an amino group in a compound of the invention.
  • the present invention includes those compounds of the invention as defined hereinbefore when made available by organic synthesis and when made available within the human or animal body by way of cleavage of a pro-drug thereof. Accordingly, the present invention includes those compounds of the invention that are produced by organic synthetic means and also such compounds that are produced in the human or animal body by way of metabolism of a precursor compound, that is a compound of the invention may be a
  • a suitable pharmaceutically acceptable pro-drug of a compound of the invention is one that is based on reasonable medical judgement as being suitable for administration to the human or animal body without undesirable pharmacological activities and without undue toxicity.
  • pro-drug Various forms of pro-drug have been described, for example in the following documents :- a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985); b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier, 1985);
  • a suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a carboxy group is, for example, an in vivo cleavable ester thereof.
  • An in vivo cleavable ester of a compound of the invention containing a carboxy group is, for example, a pharmaceutically acceptable ester which is cleaved in the human or animal body to produce the parent acid.
  • Suitable pharmaceutically acceptable esters for carboxy include
  • Ci_ 6 alkyl esters such as methyl, ethyl and ieri-butyl
  • Ci- 6 alkoxymethyl esters such as
  • Ci- 6 alkanoyloxymethyl esters such as pivaloyloxymethyl esters
  • 2-oxo-l,3-dioxolenylmethyl esters such as 5-methyl-2-oxo-l,3-dioxolen-4-ylmethyl esters and Ci ealkoxycarbonyloxy- Ci ⁇ alkyl esters such as methoxycarbonyloxymethyl and
  • a suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a hydroxy group is, for example, an in vivo cleavable ester or ether thereof.
  • An in vivo cleavable ester or ether of a compound of the invention containing a hydroxy group is, for example, a pharmaceutically acceptable ester or ether which is cleaved in the human or animal body to produce the parent hydroxy compound.
  • Suitable pharmaceutically acceptable ester forming groups for a hydroxy group include inorganic esters such as phosphate esters (including phosphoramidic cyclic esters).
  • ester forming groups for a hydroxy group include Ci ioalkanoyl groups such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups, Ci_ioalkoxycarbonyl groups such as ethoxycarbonyl, N,N -(Ci-6)2carbamoyl, 2-dialkylaminoacetyl and 2-carboxyacetyl groups.
  • Suitable pharmaceutically acceptable ether forming groups for a hydroxy group include a-acyloxyalkyl groups such as acetoxymethyl and pivaloyloxymethyl groups.
  • a suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a carboxy group is, for example, an in vivo cleavable amide thereof, for example an amide formed with an amine such as ammonia, a Ci- 4 alkylamine such as methylamine, a (Ci_ 4 alkyl) 2 amine such as dimethylamine, N-ethyl-N-methylamine or diethylamine, a Ci_ 4 alkoxy- C 2 - 4 alkylamine such as 2-methoxyethylamine, a phenyl-Ci_ 4 alkylamine such as benzylamine and amino acids such as glycine or an ester thereof.
  • an amine such as ammonia
  • a Ci- 4 alkylamine such as methylamine
  • a (Ci_ 4 alkyl) 2 amine such as dimethylamine, N-ethyl-N-methylamine or diethylamine
  • a suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses an amino group is, for example, an in vivo cleavable amide derivative thereof.
  • Suitable pharmaceutically acceptable amides from an amino group include, for example an amide formed with Ci-ioalkanoyl groups such as an acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups.
  • ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl,
  • the in vivo effects of a compound of the invention may be exerted in part by one or more metabolites that are formed within the human or animal body after administration of a compound of the invention. As stated herein before, the in vivo effects of a compound of the invention may also be exerted by way of metabolism of a precursor compound (a pro-drug).
  • compounds of invention may also be covalently linked (at any suitable position) to other groups such as, for example, solubilising moieties (for example, PEG polymers), moieties that enable them to be bound to a solid support (such as, for example, biotin-containing moieties), and targeting ligands (such as antibodies or antibody fragments).
  • solubilising moieties for example, PEG polymers
  • moieties that enable them to be bound to a solid support such as, for example, biotin-containing moieties
  • targeting ligands such as antibodies or antibody fragments
  • protecting groups see one of the many general texts on the subject, for example, 'Protective Groups in Organic Synthesis' by Theodora Green (publisher: John Wiley & Sons).
  • Protecting groups may be removed by any convenient method described in the literature or known to the skilled chemist as appropriate for the removal of the protecting group in question, such methods being chosen so as to effect removal of the protecting group with the minimum disturbance of groups elsewhere in the molecule.
  • reactants include, for example, groups such as amino, carboxy or hydroxy it may be desirable to protect the group in some of the reactions mentioned herein.
  • a suitable protecting group for an amino or alkylamino group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or i-butoxycarbonyl group, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or i-butoxycarbonyl group, an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or i-butoxycarbonyl group, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or i-butoxycarbonyl
  • arylmethoxycarbonyl group for example benzyloxycarbonyl, or an aroyl group, for example benzoyl.
  • the deprotection conditions for the above protecting groups necessarily vary with the choice of protecting group.
  • an acyl group such as an alkanoyl or
  • alkoxycarbonyl group or an aroyl group may be removed by, for example, hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide.
  • a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide.
  • an acyl group such as a teri-butoxycarbonyl group may be removed, for example, by treatment with a suitable acid as hydrochloric, sulfuric or phosphoric acid or trifluoroacetic acid and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon, or by treatment with a Lewis acid for example boron tris(trifluoroacetate).
  • a suitable alternative protecting group for a primary amino group is, for example, a phthaloyl group which may be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or with hydrazine.
  • a suitable protecting group for a hydroxy group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an aroyl group, for example benzoyl, or an arylmethyl group, for example benzyl.
  • the deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group.
  • an acyl group such as an alkanoyl or an aroyl group may be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium, sodium hydroxide or ammonia.
  • a suitable base such as an alkali metal hydroxide, for example lithium, sodium hydroxide or ammonia.
  • an arylmethyl group such as a benzyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.
  • a suitable protecting group for a carboxy group is, for example, an esterifying group, for example a methyl or an ethyl group which may be removed, for example, by hydrolysis with a base such as sodium hydroxide, or for example a i-butyl group which may be removed, for example, by treatment with an acid, for example an organic acid such as trifluoroacetic acid, or for example a benzyl group which may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.
  • a base such as sodium hydroxide
  • a i-butyl group which may be removed, for example, by treatment with an acid, for example an organic acid such as trifluoroacetic acid, or for example a benzyl group which may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.
  • Resins may also be used as a protecting group.
  • the present invention provides a process for the production of a compound of the formula I, or a pharmaceutically acceptable salt or solvate thereof, which comprises:
  • Q-L where Q has any one of the definitions set out hereinbefore and L is a leaving group; and optionally thereafter, and if necessary:
  • L is halo (e.g. fluoro, chloro or bromo, particularly chloro or bromo) or another suitable leaving group such as trifluoromethanesulfonate, Meerwein salt or methyl sulfate.
  • halo e.g. fluoro, chloro or bromo, particularly chloro or bromo
  • another suitable leaving group such as trifluoromethanesulfonate, Meerwein salt or methyl sulfate.
  • the reaction is carried out in the presence of a suitable solvent.
  • Any suitable solvent or solvent mixture may be used for this reaction.
  • a person skilled in the art will know how to select suitable solvents or solvent mixtures for use in these reactions.
  • Particular examples of suitable solvents for this reaction include dichloromethane or chloroform.
  • the reaction is carried out in a sealed vessel.
  • the reaction is carried out in the presence of a carbonate, for example, potassium carbonate.
  • a carbonate for example, potassium carbonate.
  • the reaction is carried out an elevated temperature, suitably within the range of 60 to 160 °C or, more suitably 100 to 140 °C, for a suitable time period of, for example, 6 hours to 2 days.
  • an elevated temperature suitably within the range of 60 to 160 °C or, more suitably 100 to 140 °C, for a suitable time period of, for example, 6 hours to 2 days.
  • a person skilled in the art will be able to select appropriate reaction conditions to use in order to facilitate this reaction.
  • the process defined herein may further comprise the step of subjecting the compound of formula I to a salt exchange, particularly in situations where the compound of formula I is formed as a mixture of different salt forms.
  • the salt exchange suitably comprises immobilising the compound of formula I on a suitable solid support or resin, and eluting the compounds with an appropriate acid to yield a single salt of the compound of formula I.
  • the compounds of formula A can be prepared by processes known in the art.
  • the compound of formula A can be prepared by the intra-molecular cyclisation of a compound of formula B
  • X, Ri, R2, R3, R4, R5, R 6 , R7, Rs, R21, R22 and R23 each have any one of the meanings defined hereinbefore, and L2 is a leaving group (e.g. bromine);
  • L2 is fluoro, chloro or bromo, particularly chloro or bromo or another suitable leaving group such as trifluoromthanesulfonate.
  • the intra-molecular cyclisation reaction is a palladium catalysed reaction which is suitably carried out using a strong base (such as sodium ierf-butoxide) and a ligand such as PhDavePhos (2-Diphenylphosphino-2'-(N,N-dimethylamino)biphenyl).
  • a strong base such as sodium ierf-butoxide
  • a ligand such as PhDavePhos (2-Diphenylphosphino-2'-(N,N-dimethylamino)biphenyl).
  • Any suitable solvent may be used for the cyclisation reaction.
  • suitable solvents include dimethylacetamide (DMA) and dimethylformamide (DMF).
  • reaction mixture is heated in some way.
  • the reaction is heated in a microwave for between 5 minutes and 30 minutes.
  • the compound of formula B is prepared by reacting a compound of formula C:
  • the reaction may be carried out in any suitable solvent.
  • suitable solvents include toluene and pyridine.
  • reaction is suitably carried out in the presence of a Lewis acid, such as titanium tetrachloride.
  • a Lewis acid such as titanium tetrachloride.
  • the reaction is carried out in anhydrous conditions and in the presence of an inert atmosphere, such as argon or nitrogen.
  • an inert atmosphere such as argon or nitrogen.
  • the reaction may be carried out at room temperature or at an elevated temperature, for example at the reflux temperature of the solvent used.
  • the resultant compound of formula B can be isolated and purified using techniques well known in the art.
  • a compound of formula A can be prepared by the intra-molecular cyclisation of a compound of formula E
  • X, Ri, R2, R3, R4, R5, R 6 , R7, Rs, R21, R22 and R23 each have any one of the meanings defined hereinbefore;
  • Y is halo and Z is a leaving group.
  • Y is bromo .
  • Z may be any suitable suitable leaving group.
  • Z is fluoro.
  • the intra-molecular cyclisation reaction takes place in the presence of an organolithium reagent such as t-butyl-lithium.
  • Any suitable solvent may be used for the cyclisation reaction.
  • suitable solvents include THF.
  • reaction is treated with i-butyl-lithium at low temperature.
  • the resultant compound of formula A can be isolated and purified using techniques well known in the art.
  • SPR Surface Plasmon Resonance Assay
  • Biosensor experiments were conducted in filtered, degassed HEPES buffer (10 mM HEPES, 100 mM KC1, 3 mM EDTA, 0.000 05 v/v of 10% P20 BIACORE surfactant, pH 7.3) at 25 °C.
  • the 5'-biotin labeled DNA sequences (Integrated DNA Technologies) were HPLC
  • test compounds were solubilised at lOmM in dimethyl sulfoxide (DMSO), diluted in tissue culture media and added to cells at final concentrations of 30, 10, 3, 1, 0.3, 0.1 , 0.03 and 0.01 ⁇ in triplicate. Following an incubation period of 96 hr MTT was added to a final concentration of 400 g/ml and the plates incubated for a further 4 hr. The media/MTT was then aspirated and 150 ⁇ DMSO added. Plates were shaken briefly and the absorbance at 550nm read using a Labtech LT-4000 plate reader. An extra plate was seeded and the MTT assay was carried out on this plate at the time of drug addition to ascertain an initial absorbance value.
  • DMSO dimethyl sulfoxide
  • the compounds of formula I demonstrate a quadraplexrduplex ratio in the SPR assay of greater than 2.
  • the quadraplex: duplex ratio in the SPR assay of the compound of Example 2 herein is greater than 15.
  • antiproliferative activity possessed by compounds of the formula I may be demonstrated in the growth inhibition assay by a GI 50 value of less than 40 ⁇ (preferred compounds have an GI 50 of less than 15 ⁇ , more preferably less than 10 ⁇ ).
  • the compound is not one the above-identified compounds.
  • Pharmaceutical Compositions are not one the above-identified compounds.
  • composition which comprises a compound of the invention as defined hereinbefore, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable diluent or carrier.
  • compositions of the invention may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular, intraperitoneal or intramuscular dosing or as a suppository for rectal dosing).
  • oral use for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or
  • compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art.
  • compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and/or preservative agents.
  • An effective amount of a compound of the present invention for use in therapy of infection is an amount sufficient to symptomatically relieve in a warm-blooded animal, particularly a human the symptoms of infection, to slow the progression of infection, or to reduce in patients with symptoms of infection the risk of getting worse.
  • a formulation intended for oral administration to humans will generally contain, for example, from 0.5 mg to 0.5 g of active agent (more suitably from 0.5 to 100 mg, for example from 1 to 30 mg) compounded with an appropriate and convenient amount of excipients which may vary from about 5 to about 98 percent by weight of the total composition.
  • the size of the dose for therapeutic or prophylactic purposes of a compound of the invention will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine.
  • a daily dose in the range for example, 0.1 mg/kg to 75 mg/kg body weight is received, given if required in divided doses.
  • a parenteral route is employed.
  • a dose in the range for example, 0.1 mg kg to 30 mg kg body weight will generally be used.
  • a dose in the range for example, 0.05 mg/kg to 25 mg kg body weight will be used.
  • Oral administration may also be suitable, particularly in tablet form.
  • unit dosage forms will contain about 0.5 mg to 0.5 g of a compound of this invention.
  • the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for use as a medicament.
  • the compounds of the invention are capable of inhibiting telomerase activity.
  • the present invention provides a method of inhibiting telomerase activity in a cell, the method comprising administering to said cell compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for the inhibition of telomerase in a cell (in vivo or ex vivo) .
  • the present invention provides a method of inhibiting telomerase activity in a human or animal subject in need of such inhibition, the method comprising administering to said subject an effective amount of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof for use in the treatment of disease or condition associated with telomerase activity.
  • the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for use in the treatment of disease or condition associated with telomerase activity.
  • the present invention provides a method of treating a proliferative disorder in a human or animal subject, the method comprising administering to said subject a therapeutically acceptable amount of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for use in the treatment of a proliferative disorder.
  • the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for use in the treatment of a proliferative disorder.
  • proliferative disorder are used interchangeably herein and pertain to an unwanted or uncontrolled cellular proliferation of excessive or abnormal cells which is undesired, such as, neoplastic or hyperplastic growth, whether in vitro or in vivo.
  • proliferative conditions include, but are not limited to, pre-malignant and malignant cellular proliferation, including but not limited to, malignant neoplasms and tumours, cancers, leukemias, psoriasis, bone diseases, fibroproliferative disorders (e.g., of connective tissues), and
  • any type of cell may be treated, including but not limited to, lung, colon, breast, ovarian, prostate, liver, pancreas, brain, and skin.
  • the anti-proliferative effects of the compounds of the present invention have particular application in the treatment of human cancers by virtue of their telomerase inhibitory properties.
  • the anti-cancer effect may arise through one or more mechanisms, including but not limited to, the regulation of cell proliferation, the inhibition of angiogenesis (the formation of new blood vessels), the inhibition of metastasis (the spread of a tumour from its origin), the inhibition of invasion (the spread of tumour cells into neighbouring normal structures), or the promotion of apoptosis (programmed cell death).
  • the invention further provides a method of treatment of the human or animal body, the method comprising administering to a subject in need of treatment a therapeutically-effective amount of an active compound, preferably in the form of a pharmaceutical composition.
  • Telomerase targeting agents are also known to be useful for inhibiting cancer stem cells (see, for example, US2008/0279961).
  • the present invention also provides a method of inhibiting a cancer stem cell, the method comprising administering to said cell an effective amount of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for use in the inhibition and/or treatment of cancer stem cells.
  • the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for use in the inhibition and/or treatment of cancer stem cells.
  • the compounds of the invention or pharmaceutical composition comprising the active compound may be administered to a subject by any convenient route of administration, whether systemically/ peripherally or topically (ie., at the site of desired action).
  • Routes of administration include, but are not limited to, oral (e.g, by ingestion); buccal; sublingual; transdermal (including, e.g., by a patch, plaster, etc.); transmucosal (including, e.g., by a patch, plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by eyedrops); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., via an aerosol, e.g., through the mouth or nose); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, infraarterlal, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratrache
  • the antiproliferative treatment defined hereinbefore may be applied as a sole therapy or may involve, in addition to the compound of the invention, conventional surgery or radiotherapy or chemotherapy.
  • Such chemotherapy may include one or more of the following categories of anti-tumour agents :-
  • alkylating agents for example cis-platin, oxaliplatin, carboplatin,
  • cyclophosphamide nitrogen mustard, melphalan, chlorambucil, busulphan, temozolamide and nitrosoureas
  • antimetabolites for example gemcitabine and antifolates such as
  • fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea
  • antitumour antibiotics for example anthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin
  • antimitotic agents for example vinca alkaloids like vincristine, vinblastine, vindesine and vinorelbine and taxoids like taxol and taxotere and polokinase inhibitors
  • topoisomerase inhibitors for example irinotecan, etoposide, teniposide, amsacrine, topotecan and camptothecin
  • PARP inhibitors for example irinotecan, etoposide, teniposide, amsacrine, topotecan and camptothecin
  • irinotecan for example irinotecan, etoposide, teniposide, amsacrine, topotecan and camptothecin
  • PARP inhibitors for example irinotecan, etoposide, teniposide, amsacrine, topotecan and camptothecin
  • cytostatic agents such as antioestrogens (for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene), antiandrogens (for example bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin), progestogens (for example megestrol acetate), aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5a- reductase such as finasteride;
  • antioestrogens for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene
  • antiandrogens for example
  • anti-invasion agents for example c-Src kinase family inhibitors like 4-(6-chloro-2,3- methylenedioxyanilino)-7-[2-(4-methylpiperazin-l-yl)ethoxy]-5-tetrahydropyran-4- yloxyquinazoline (AZD0530; International Patent Application WO 01/94341), N-(2-chloro-6- methylphenyl)-2- ⁇ 6-[4-(2-hydroxyethyl)piperazin-l-yl]-2-methylpyrimidin-4-ylamino ⁇ thiazole- 5-carboxamide (dasatinib, BMS-354825; J. Med.
  • anti-invasion agents for example c-Src kinase family inhibitors like 4-(6-chloro-2,3- methylenedioxyanilino)-7-[2-(4-methylpiperazin-l-yl)ethoxy]-5-tetrahydropyr
  • inhibitors of growth factor function include growth factor antibodies and growth factor receptor antibodies (for example the anti-erbB2 antibody trastuzumab [HerceptinTM] , the anti-EGFR antibody panitumumab, the anti-erbBl antibody cetuximab [Erbitux, C225] and any growth factor or growth factor receptor antibodies disclosed by Stern et al. Critical reviews in oncology/haematology, 2005, Vol.
  • inhibitors also include tyrosine kinase inhibitors, for example inhibitors of the epidermal growth factor family (for example EGFR family tyrosine kinase inhibitors such as N-(3-chloro-4- fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, ZD1839), N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) and 6- acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib); inhibitors of the hepatocyte growth factor family; inhibitors of the insulin growth factor family; inhibitors of the insulin growth
  • antiangiogenic agents such as those which inhibit the effects of vascular endothelial growth factor, [for example the anti-vascular endothelial cell growth factor antibody bevacizumab (AvastinTM) and for example, a VEGF receptor tyrosine kinase inhibitor such as vandetanib (ZD6474), vatalanib (PTK787), sunitinib (SU11248), axitinib (AG-013736), pazopanib (GW 786034) and 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-l- ylpropoxy)quinazoline (AZD2171; Example 240 within WO 00/47212), compounds such as those disclosed in International Patent Applications W097/22596, WO 97/30035, WO 97/32856 and WO 98/13354 and compounds that work by other mechanisms (for example linomide
  • vascular damaging agents such as Combretastatin A4 and compounds disclosed in International Patent Applications WO 99/02166, WO 00/40529, WO 00/41669,
  • an endothelin receptor antagonist for example zibotentan (ZD4054) or atrasentan;
  • antisense therapies for example those which are directed to the targets listed above, such as ISIS 2503, an anti-ras antisense;
  • (ix) gene therapy approaches including for example approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT (gene-directed enzyme pro-drug therapy) approaches such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multi-drug resistance gene therapy; and
  • GDEPT gene-directed enzyme pro-drug therapy
  • (x) immunotherapy approaches including for example ex-vivo and in-vivo approaches to increase the immunogenicity of patient tumour cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor, approaches to decrease T-cell anergy, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumour cell lines and approaches using anti-idiotypic antibodies.
  • cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor
  • Particular combinations of interest include the compounds of the invention with a topioisomerase inhibitor (such as irinotecan) and/or a PARP inhibitor.
  • Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment.
  • Such combination products employ the compounds of this invention within the dosage range described hereinbefore and the other pharmaceutically- active agent within its approved dosage range.
  • a combination suitable for use in the treatment of a cancer comprising a compound of the invention as defined hereinbefore, or a pharmaceutically acceptable salt or solvate thereof, and another anti-tumour agent.
  • a combination suitable for use in the treatment of a cancer comprising a compound of the invention as defined hereinbefore, or a pharmaceutically acceptable salt or solvate thereof, and any one of the anti-tumour agents listed under (i) - (ix) above.
  • a pharmaceutical composition which comprises a compound of the invention, or a pharmaceutically acceptable salt or solvate thereof in combination with an anti-tumour agent selected from one listed under (i) - (ix) herein above, in association with a pharmaceutically acceptable diluent or carrier.
  • LCMS data were obtained using an Agilent 1200 series LC/MS with an Agilent 6110 quadropole MS, with Electrospray ionisation. The following three methods were used:
  • Mobile phase A 0.1% Acetic acid in water.
  • Mobile Phase B 0.1% acetic acid in acetonitrile. Flow rate of l.OOml/min. Gradient from 2% B over 3 minutes to 20%B for 0.2 minutes, up to 95% B for 1.8 minutes and then back down to 2%B. Total run time of 6 minutes.
  • Mass Spectra were achieved using the MS in positive mode.
  • Mobile phase A 0.1% ammonia solution in water.
  • Mobile phase B 0.1% ammonia solution in acetonitrile. Flow rate of 0.500ml/min. Gradient from 5% B over 3 minutes to 95%B, remaining at 95%B until 4.1 minutes, when it returns down to 5%B until the 7 minute run is over.
  • Mass Spectra were achieved using the MS in positive mode.
  • reversed-phase silica was used with a flow rate of about 1 ml per minute and detection was by Electrospray Mass Spectrometry and by UV absorbance at a wavelength of 254 nm.
  • Silica bound trimethylammonium chloride resin (1 g) was stirred in degassed water and was filtered. This process was repeated three times and the resin was added to 1L of a 0.1M aqueous solution of 2:1 methanesulfonic acid: sodium methanesulfonate. The mixture was stirred for 10 min. and decantated. This was repeated two more times and the silica was filtered and transferred to a small column.
  • silica bound resin was washed successively with two column volume of 0.1M aqueous solution of 2:1 methanesulfonic acid: sodium methanesulfonate, two column volume of 0.01M aqueous solution of 2:1 methanesulfonic acid: sodium methanesulfonate, and two column volume of water to give silica bound trimethyl ammonium mesylate.
  • Acetic anhydride (3.24 mL, 34.3 mmol, 1.1 equ) was added dropwise to a solution of propargyl amine (2 mL, 31.2 mmol), pyridine (2.76 mL, 34.3 mmol, 1.1 equ) and N,N dimethylaminopyridine (few crystals, catalytic amount) in dichloromethane (50 mL) at 0°C and the mixture was stirred overnight. The mixture was diluted with dichloromethane and transferred into a separating funnel. Water was added and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate. The product was absorbed on silica and purified by flash chromatography (Hexane:EtOAc 30%-20 ) to give the title compound as an off white crystalline solid (1.45 g, 49% yield).
  • An air condenser (50 cm length) loosely packed with glass wool was attached to the reaction vessel. The outlet of the air condenser was connected to a Dewar condenser filled with dry ice-acetone connected to a receiving flask (250 mL) cooled in a dry ice-acetone bath.
  • the receiving flask was also connected in series to a second dry ice-acetone cold trap, a trap containing potassium iodide solution, and a vacuum was placed on the apparatus.
  • Oxone was added in portions (10-15 g) over approximately 0.5h while the acetone-water mixture was simultaneously added dropwise and the reaction mixture was stirred vigorously throughout the addition of reagents.
  • a yellow solution of 3,3-dimethyldioxirane in acetone collected in the receiving flask. Vigorous stirring was continued for an additional 15 min while a vacuum was applied to the cold trap.
  • the yellow dioxirane solution was dried over sodium sulfate, filtered and stored in the freezer (-25 °C) over activated molecular sieves.
  • the overall yield was 65mL.
  • a microwave vial was charged with 2-bromo-N-(9H-xanthen-9-ylidene)aniline (250 mg, 1.43 mmol), sodium ie/t-butoxide (274 mg, 2.86 mmol, 2.0 equ), a catalytic amount of tris(dibenzylideneacetone)dipalladium, followed by a catalytic amount of diphenylphosphino-2- (N,N-dimethylamino)biphenyl.
  • the RM was diluted with anhydrous dimethylacetamide (5 mL) and then heated under microwave radiation at 180°C (300W, 200psi, 5 mins). This reaction was repeated in duplicate.
  • Step 1 Preparation of 2',3-difluoro-4'-methoxybiphenyl-2-carbonitrile
  • N,N- diisopropylethylamine 100 mg, 0.76 mmol, 2 equ
  • N-phenyl bis-trifluoromethane sulfonimide 207 mg, 0.58 mmol, 1.5 equ
  • the reaction mixture was then evaporated to dryness, and purified by flash chromatography (chloroform) to give the title compound as pale yellow solid (100 mg, 62% yield).
  • reaction mixture was concentrated in vacuo, dissolved in 95% DCM:MeOH and purified by flash column chromatography (gradient elution DCM:MeOH 98%-90%). The combined pure fractions were concentrated in vacuo, and dissolved in DCM:MeOH 90% (10 mL). This was stirred with silica bound trimethylammonium- mesylate (400 mg) for 2 h and filtered through Celite. The filtrate was evaporated to dryness to give the title compound as a yellow solid (39 mg, 0.0875 mmol, 36% yield).
  • a mixture 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.240 mmol; Example 4), tetrakis(triphenylphosphine)palladium(0) (27 mg, 0.0240 mmol, 0.1 equ), potassium carbonate (67 mg, 0.480 mmol, 2 equ) and 3-furanyl boronic acid (54 mg, 0.480 mmol, 2 equ) in 1 :1 DME:H 2 0 (6 mL) was heated at 80°C under microwave irradiation (300 W, 300 psi) for 10 min.
  • the reaction mixture was concentrated in vacuo, dissolved in 95% DCM:MeOH and purified by flash column chromatography (gradient elution DCM:MeOH 98%-90%). The combined pure fractions were concentrated in vacuo, dissolved in DCM:MeOH 90% (10 mL) and stirred with silica bound trimethylammonium-mesylate (750 mg) for 2 h. The mixture was filtered through Celite and the filtrate was evaporated to dryness to give the title compound as a yellow solid (86 mg, 0.193 mmol, 80% yield).
  • reaction mixture was dissolved in DCM:MeOH 90% (8 mL) and purified by column chromatography (gradient elution DCM:MeOH 90%-80%). The combined pure fractions were concentrated in vacuo, dissolved in 90% DCM:MeOH (10 mL) and was stirred with silica bound trimethylammonium-mesylate (1.5 g) for 2 h. The mixture was filtered through Celite and the filtrate was evaporated to dryness in vacuo to give the title compound as a yellow solid (145 mg, 0.330 mmol, 91% yield).
  • 2-Bromo-5-nitroaniline (4.43 g, 20.4 mmol, 2 equ) was added portionwise to a stirring solution of xanthone (2.00 g, 10.2 mmol), 1.0 M titanium chloride in toluene (10.2 mL, 10.2 mmol, 1 equ) and N,N-diisopropylethylamine (13.2 g, 102 mmol, 10 equ) in toluene (300 mL). The mixture was refluxed under nitrogen for 2 h and 1.0 M titanium chloride in toluene (10.2 mL, 10.2 mmol, 1 equ) was added. The reaction mixture was refluxed under nitrogen for 16 h and was allowed to cool to room temperature.
  • Phthaloyl dichlonde (88 mg, 0.422 mmol, 1.5 equ) was added to a stirring solution of chromeno[4,3,2-g/z]phenanthridin-2-amine (80 mg, 0.282 mmol), pyridine (112 mg, 1.408 mmol, 5 equ) and N, N dimethylaminopyridine ( ⁇ 1 mg, catalytic amount) in dichloromethane (40 mL).
  • the reaction mixture was allowed to stir under nitrogen at room temperature for 10 min and then concentrated in vacuo.
  • the solid residue was triturated with 0.5 M HC1 (aq) (20 mL) and the resulting precipitate was collected by filtration.
  • Acetyl chloride (22 mg, 0.280 mmol, 6.3 equ) was added to a stirring solution of 2-amino-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate (20 mg, 0.0446 mmol) and pyridine (24 mg, 0.281 mmol, 6.3 equ) in dichloromethane (2 mL) for 1 h at room temperature.
  • the reaction mixture was evaporated to dryness in vacuo, dissolved in DCM/MeOH 90% (3 mL) and purified by flash column chromatography (gradient elution 90%- 85%).
  • Acetyl chloride (8 mg, 0.0983 mmol) was added to the mixture of 13-methyl-2- (methylamino)chromeno[4,3,2- z]phenanthridin-13-ium trifluoromethanesulfonate and 2- (dimethylamino)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate obtained above (21 mg) and pyridine (11 mg, 0.131 mmol) in dichloromethane (2 mL) and stirred under nitrogen at room temperature for 1 h.
  • reaction mixture was concentrated in vacuo and dissolved in DCM:MeOH 90% (5 mL) and purified by flash column chromatography (gradient elution DCM:MeOH 90%-86%) giving two separate product fractions.
  • the first combined pure fractions were concentrated in vacuo to give 13-mefhyl-2-(N- methylacetamido)chromeno[4,3,2-g/z]phenanthridin-13-ium 1/3 trifluoromethanesulfonate 2/3 acetate as a yellow solid (10 mg, 0.0198 mmol).
  • 2-acetamido- 13-methylchromeno[4,3,2-gA]phenanthridin- 13-ium methanesulfonate could be obtained by palladium catalysed cross-coupling of acetamide with 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2- z]phenanthridin-13-ium
  • a second portion of caesium carbonate (14 mg, 0.240 mmol, 3 equ) was added and the mixture was heated at 80 °C under microwave irradiation (300 W, 300 psi) for 5 min.
  • the reaction mixture was filtered through Celite, evaporated to dryness in vacuo, dissolved in DCM:MeOH 95% (4 mL) and purified by flash column chromatography (gradient elution DCM:MeOH 95%-90%). The combined pure fractions were concentrated in vacuo and the resulting orange solid was then stirred in DCM:MeOH (90%, 20 ml) when polymer supported trialkylammonium mesylate resin (0.500 g) was added.
  • Acetyl chloride (4.6 ⁇ L, 0.0646 mmol, 1.5 equ) was added to a suspension of 2-amino-13- methylthiochromeno[4,3,2- z]phenanthridin-13-ium trifluoromethanesulfonate (20 mg, 0.04431 mmol) and N,N-Diisopropylethylamine (9 ⁇ L , 0.0517 mmol, 1.2 equ) in dichloromethane (1 mL) at 0°C and the mixture was stirred at 0°C for 45 min.
  • the mixture was basified at 0°C to pH 13 with 5N NaOH and a saturated solution of potassium sulfite was added.
  • the product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate then concentrated in vacuo.
  • the product was dissolved in dichloromethane and the solution was loaded on a short column packed with silica.
  • the product was eluted with DCM to give the title compound as a yellow/pale brown solid (75 mg, 69% yield).
  • Step 1 Preparation of 3, 6-dihydroxy-9H-xanthen-9-one (synthesized by modification of a procedure reported in Grover, P. K.; Shah, G. D.; Shah, R. C. /. Chem. Soc. 1955, 3982-3985)
  • a suspension of bis(2,4-dihydroxyphenyl)methanone (6 g, 24.4 mmol) in water was heated at 190°C for 48 hours.
  • the reaction mixture was cooled to room temperature and the resulting precipitate was filtered, washed with acetonitrile and diethyl ether to give the title compound as pale yellow solid (5.56 g, 100% yield).
  • Step 2 Preparation of 6,10-bis(2-hydroxyethoxy)-13-methylchromeno[4,3,2- gft]phenanthridin-13-ium bromide
  • the mixture was stirred at 100°C for 2 h and was allowed to cool to room temperature.
  • the mixture was diluted with N,N-dimethylformamide and filtered through a syringe filter.
  • the filtrate was concentrated in vacuo and the product was dissolved in DCM:MeOH.
  • the product was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 95%-90 -85%) to give the title compound as a yellow/orange solid (13 mg, 54% yield).
  • reaction mixture was quenched with methanol and filtered; the filtrate was concentrated to dryness and the solid residue was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 100%-95%) to give the title compound as a solid (13 mg, 45% yield).
  • Step 1 Preparati -difluoro-4-methoxyphenylboronic acid
  • Methyl iodide (1.56 mL, 25.1 mmol, 1.5 equ) was added dropwise to a suspension of 4-bromo- 2,3-difluorophenol (3.5 g, 16.7 mmol) and potassium carbonate (2.77 g, 20.0 mmol, 1.2 equ) in N,N-dimethylformamide (30 mL) and the mixture was stirred for 3 days. The precipitate was removed by filtration and the filtrate was concentrated to dryness in vacuo. Dichloromethane and water were added and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate and concentrated in vacuo to give l-bromo-2,3- difluoro-4-methoxybenzene as a white crystalline solid (3.8 g, quantitative).
  • Nitrogen was bubbled through a suspension of 2-bromo-6-fhiorobenzonitrile (2.39 g, 12.0 mmol), 2,3-difluoro-4-methoxyphenylboronic acid (2.7 g, 14.4 mmol, 1.2 equ) and potassium fluoride (2.3 g, 39.6 mmol, 3.3 equ) in tetrahydrofuran (45 mL) for 10 min. before the addition of bis(tri-iert-butylphosphine)palladium (123 mg, 0.24 mmol, 0.02 equ). The mixture was immersed into a pre-heating oil bath at 55°C and the mixture was stirred at 55°C for 45 min.
  • reaction was quenched with acetic acid and 1M NaOH, followed by sat. NaHCC>3 were added.
  • the resulting bright yellow suspension was transferred into a separating funnel and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate.
  • a reaction mixture from a separate experiment carried out on 85 mg scale was combined and the mixture was absorbed on silica.
  • the product was purified by flash chromatography (gradient elution CHCl 3 :Hexane 25%-50%-100% then DCM:MeOH 97.5%- 95%) to give the title compound as a yellow solid (565 mg, 50% yield).
  • the product could be only partially purified as some impurities co-eluted with the product during the purification by flash chromatography. The product was used without further purification in the next steps.
  • Step 5 Preparation of l,ll-difluoro-2-methoxy-13-methylchromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate
  • Methyl triflate (98 iL, 0.895 mmol, 10 equ ) was added to a suspension of l,l l-difluoro-2- methoxychromeno[4,3,2-g/z]phenanthridine (30 mg, 0.0895 mmol) and potassium carbonate (124 mg, 0.895 mmol, 10 equ) in chloroform (3 mL) and the mixture was heated at 110°C m a sealed tube overnight. The mixture was cooled to room temperature and the reaction was quenched with methanol. The mixture was stirred for ⁇ 30min. and was filtered.
  • Step 1 Preparation of l,ll-difluorochromeno[4,3,2- ⁇ ]phenanthridin-2-yl
  • Step 2 Preparation of l,ll-difluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2- g/i]phenanthridin-13-ium trifluoromethanesulfonate
  • Methyl triflate (532 ⁇ , 4.85 mmol, 10 equ) was added to a suspension of 1,11- difluorochromeno[4,3,2-g3 ⁇ 4phenanthridin-2-yl trifluoromethanesulfonate (220 mg, 0.485 mmol) and potassium carbonate (670 mg, 4.85 mmol, 10 equ) in chloroform (5 mL) and the mixture was heated in a sealed tube at 100°C overnight. The mixture was cooled to room temperature and methanol was added. The precipitate was removed by filtration and the filtrate was absorbed on silica. The product was purified by flash chromatography (gradient elution DCM:MeOH 100%- 98%-97%-96%-92%) to give l,l l-difluoro-13-methyl-2-
  • the resulting solid was dissolved in DCMrMeOH, absorbed on silica and purified by flash chromatography (gradient elution DCMrMeOH 100%-98%-97%-96%-94%-92%-90%) to give 26 mg of an orange solid.
  • the material was triturated with 3N HCl, filtered, washed successively with water and diethyl ether, then dried under suction to give the title compound as an orange solid (15 mg, 24% yield).

Abstract

The present invention relates to novel compounds of formula (I) wherein X, Q, R1, R2, R3, R4, R5, R6, R7, R8, R21, R22 and R23 are as defined herein. The compounds of formula (I) are inhibitors of telomerase enzyme function and are accordingly useful for the treatment cellular proliferation disorders, such as cancer.

Description

FUSED PENTACYCLIC ANTI - PROLIFERATIVE COMPOUNDS
FIELD OF THE INVENTION
[0001] The present invention relates to novel compounds that function as inhibitors of the activity of the enzyme telomerase, either directly or indirectly via interaction with the telomere itself. More specifically, the present invention relates to novel compounds which are potentially useful therapeutic agents for the treatment and/or prevention of proliferative diseases, such as cancer. The present invention also relates to processes for the preparation of these compounds, to pharmaceutical compositions comprising them and to their therapeutic uses.
BACKGROUND OF THE INVENTION
[0002] Cancer is caused by uncontrolled and unregulated cellular proliferation. Precisely what causes a cell to become malignant and proliferate in an uncontrolled and unregulated manner has been the focus of intense research over recent decades. This research has led to the identification of numerous tumour- specific or tumour-associated targets, which can then be targeted by anticancer agents.
[0003] One particular therapeutic target that has attracted particular attention in the cancer field is the telomerase enzyme, which catalyses the synthesis of telomeres.
[0004] Telomeres are nucleoprotein structures at the ends of linear chromosomes which consist of DNA sequences arranged in tandemly repeated units which extend from less than 100 to several thousands of bases. With each cell division cycle, bases are lost from the ends of the chromosomes and, as these telomeres shorten in length, the cell eventually reaches a point at which apoptosis is triggered.
Telomerase is a ribonucleoprotein reverse transcriptase that functions to maintain the telomere length above the point at which apoptosis is triggered by utilising its internal RNA template to synthesize telomeric DNA sequences, which compensate for the loss of telomeric DNA (see Blackburn; Annu. Rev.
Biochem.; 1992; 61:113-129). This prevents further shortening of the telomeres, and the resulting stabilization of their length contributes to immortalisation.
[0005] Telomerase is present in stem and germ line cells of normal tissues, but is absent in most other cells. In addition, telomerase activity, and the associated shortened but stabilized telomeres, have been detected in the majority of tumours examined (and in over 90% of all human cancers examined). As a consequence, the telomerase enzyme is a particularly interesting target for anti-cancer therapy. Furthermore, the absence of telomerase in most normal cells makes this enzyme a particularly attractive target because its inhibition would be expected to cause minimal damage to normal healthy cells and tissues. The fact that tumour cells have shorter telomeres and higher proliferation rates than normal replicative cell populations suggests that a therapeutic telomerase inhibitor may cause tumour cell death well before damage to regenerative tissues occurs, thereby minimising undesirable side effects.
[0006] A more detailed discussion of telomeres and telomerase, and their role as anti-proliferative targets is provided in, for example, Sharma et al, Annals of Oncology, 1997, Vol. 8, pp 689-685; Urquidi et al, Ann. Med., 1998, Vol. 30, pp 419-430; Perry etal, Exp. Opin. Ther. Patents, 1998, Vol. 8, No. 12, pp 1567-1586; Autexier, Chemistry & Biology, 1999, Vol. 6, pp R299-R303; andNeidle i¾a/., Anti-Cancer Drug Design, 1999, Vol. 14, pp 341-347.
[0007] Certain compounds that target the telomerase enzyme are known. For example, WO 02/30932 describes certain N8, N13-disubstituted quino[4,3,2-KL]acridium salts that function as inhibitors of telomerase.
[0008] However, there still remains a need for alternative and improved telomerase inhibitors. In particular, there remains a need for telomerase inhibitors that possess one or more advantageous pharmaceutical properties and which are potentially useful therapeutic agents for the treatment of proliferative disorders such as cancer. In addition, there remains a need for improved telomerase inhibitors which exhibit good selectivity for the target as well as reduced non-target activity.
[0009] It is therefore an object of the present invention to provide alternative and improved telomerase inhibitors that can be used as pharmaceutical agents for use in the treatment of proliferative disorders such as cancer.
SUMMARY OF THE INVENTION
[0010] In a first aspect, the present invention provides a compound as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
[0011] In another aspect, the present invention provides a pharmaceutical composition comprising a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein, in admixture with a pharmaceutically acceptable diluent or carrier.
[0012] In another aspect, the present invention provides a method of inhibiting telomerase in vitro or in vivo, said method comprising contacting a cell with an effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein. [0013] In another aspect, the present invention provides a method of inhibiting cell proliferation in vitro or in vivo, said method comprising contacting a cell with an effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein.
[0014] In another aspect, the present invention provides a method of treating a proliferative disorder in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein.
[0015] In a more particular aspect, the present invention provides a method of treating cancer in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein.
[0016] In another aspect, the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein for use in therapy.
[0017] In another aspect, the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of a proliferative condition.
[0018] In another aspect, the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of cancer. In a particular embodiment, the cancer is a human cancer.
[0019] In another aspect, the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein for use in the production of a telomerase inhibitory effect.
[0020] In another aspect, the present invention provides the use of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein in the manufacture of a medicament for use in the treatment of a proliferative condition.
[0021] In another aspect, the present invention provides the use of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein in the manufacture of a medicament for use in the treatment of cancer. Suitably, the medicament is for use in the treatment of human cancers.
[0022] In another aspect, the present invention provides the use of a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein in the manufacture of a medicament for use in the production of a telomerase inhibitory effect. [0023] The present invention further provides a method of synthesising a compound, or a pharmaceutically acceptable salt or solvate thereof, as defined herein.
[0024] In another aspect, the present invention provides a compound, or a pharmaceutically acceptable salt or solvate thereof, obtainable by, or obtained by, or directly obtained by a method of synthesis as defined herein.
[0025] In another aspect, the present invention provides novel intermediates as defined herein which are suitable for use in any one of the synthetic methods set out herein.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0026] Unless otherwise stated, the following terms used in the specification and claims have the following meanings set out below.
[0027] It is to be appreciated that references to "treating" or "treatment" include prophylaxis as well as the alleviation of established symptoms of a condition. "Treating" or "treatment" of a state, disorder or condition therefore includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in a human that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition, (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical or subclinical symptom thereof, or (3) relieving or attenuating the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms.
[0028] A "therapeutically effective amount" means the amount of a compound that, when administered to a mammal for treating a disease, is sufficient to effect such treatment for the disease. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity and the age, weight, etc., of the mammal to be treated.
[0029] In this specification the term "alkyl" includes both straight and branched chain alkyl groups. References to individual alkyl groups such as "propyl" are specific for the straight chain version only and references to individual branched chain alkyl groups such as "isopropyl" are specific for the branched chain version only. For example, "(l-6C)alkyl" includes (l-4C)alkyl, (l-3C)alkyl, propyl, isopropyl and i-butyl. A similar convention applies to other radicals, for example "phenyl(l-6C)alkyl" includes phenyl(l-4C)alkyl, benzyl, 1-phenylethyl and 2-phenylethyl.
[0030] The term "(m-nC)" or "(m-nC) group" used alone or as a prefix, refers to any group having m to n carbon atoms.
[0031] An "alkylene," "alkenylene," or "alkynylene" group is an alkyl, alkenyl, or alkynyl group that is positioned between and serves to connect two other chemical groups. Thus, "(1- 6C)alkylene" means a linear saturated divalent hydrocarbon radical of one to six carbon atoms or a branched saturated divalent hydrocarbon radical of three to six carbon atoms, for example, methylene, ethylene, propylene, 2-methylpropylene, pentylene, and the like.
[0032] "(2-6C)alkenylene" means a linear divalent hydrocarbon radical of two to six carbon atoms or a branched divalent hydrocarbon radical of three to six carbon atoms, containing at least one double bond, for example, as in ethenylene, 2,4-pentadienylene, and the like.
[0033] "(2-6C)alkynylene" means a linear divalent hydrocarbon radical of two to six carbon atoms or a branched divalent hydrocarbon radical of three to six carbon atoms, containing at least one triple bond, for example, as in ethynylene, propynylene, and butynylene and the like.
[0034] "(3-8C)cycloalkyl" means a hydrocarbon ring containing from 3 to 8 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or bicyclo[2.2.1]heptyl.
[0035] "(3-8C)cycloalkenyl" means a hydrocarbon ring containing at least one double bond, for example, cyclobutenyl, cyclopentenyl, cyclohexenyl or cycloheptenyl, such as 3-cyclohexen-l- yl, or cyclooctenyl.
[0036] "(3-8C)cycloalkyl-(l-6C)alkylene" means a (3-8C)cycloalkyl group covalently attached to a (l-6C)alkylene group, both of which are defined herein.
[0037] The term "halo" refers to fluoro, chloro, bromo and iodo.
[0038] The term "heterocyclyl", "heterocyclic" or "heterocycle" means a non-aromatic saturated or partially saturated monocyclic, fused, bridged, or spiro bicyclic heterocyclic ring system(s). The term heterocyclyl includes both monovalent species and divalent species. Monocyclic heterocyclic rings contain from about 3 to 12 (suitably from 3 to 7) ring atoms, with from 1 to 5 (suitably 1, 2 or 3) heteroatoms selected from nitrogen, oxygen or sulfur in the ring. Bicyclic heterocycles contain from 7 to 17 member atoms, suitably 7 to 12 member atoms, in the ring. Bicyclic heterocyclic(s) rings may be fused, spiro, or bridged ring systems. Examples of heterocyclic groups include cyclic ethers such as oxiranyl, oxetanyl, tetrahydrofuranyl, dioxanyl, and substituted cyclic ethers. Heterocycles containing nitrogen include, for example, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, tetrahydrotriazinyl, tetrahydropyrazolyl, and the like. Typical sulfur containing heterocycles include tetrahydrothienyl, dihydro-l ,3-dithiol, tetrahydro- 2H-thiopyran, and hexahydrothiepine. Other heterocycles include dihydro-oxathiolyl, tetrahydro-oxazolyl, tetrahydro-oxadiazolyl, tetrahydrodioxazolyl, tetrahydro-oxathiazolyl, hexahydrotriazinyl, tetrahydro-oxazinyl, morpholinyl, thiomorpholinyl, tetrahydropyrimidinyl, dioxolinyl, octahydrobenzofuranyl, octahydrobenzimidazolyl, and octahydrobenzothiazolyl. For heterocycles containing sulfur, the oxidized sulfur heterocycles containing SO or S02 groups are also included. Examples include the sulfoxide and sulfone forms of tetrahydrothienyl and fhiomorpholinyl such as tetrahydrothiene 1,1 -dioxide and thiomorpholinyl 1,1 -dioxide. A heterocyclyl gourp may optionally bear 1, 2 or more oxo (=0) or thioxo (=S) substituents. A suitable value for a heterocyclyl group which bears 1 or 2 oxo (=0) or thioxo (=S) substituents is, for example, 2-oxopyrrolidinyl, 2-thioxopyrrolidinyl, 2-oxoimidazolidinyl, 2-thioxoimidazolidinyl, 2-oxopiperidinyl, 2,5-dioxopyrrolidinyl, 2,5-dioxoimidazolidinyl or 2,6-dioxopiperidinyl. Particular heterocyclyl groups are saturated monocyclic 3 to 7 membered heterocyclyls containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen or sulfur, for example azetidinyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, tetrahydrothienyl, tetrahydrothienyl 1,1 -dioxide, thiomorpholinyl, thiomorpholinyl 1,1 -dioxide, piperidinyl, homopiperidinyl, piperazinyl or homopiperazinyl. As the skilled person would appreciate, any heterocycle may be linked to another group via any suitable atom, such as via a carbon or nitrogen atom. However, reference herein to piperidino or morpholino refers to a piperidin-l-yl or morpholin-4-yl ring that is linked via the ring nitrogen.
[0039] By "bridged ring systems" is meant ring systems in which two rings share more than two atoms, see for example Advanced Organic Chemistry, by Jerry March, 4th Edition, Wiley Interscience, pages 131-133, 1992. Examples of bridged heterocyclyl ring systems include, aza- bicyclo[2.2.1]heptane, 2-oxa-5-azabicyclo[2.2.1]heptane, aza-bicyclo[2.2.2]octane, aza- bicyclo [3.2.1] octane and quinuclidine.
[0040] "Heterocyclyl(l-6C)alkyl" means a heterocyclyl group covalently attached to a (1- 6C)alkylene group, both of which are defined herein.
[0041] The term "heteroaryl" or "heteroaromatic" means an aromatic mono-, bi-, or polycyclic ring incorporating one or more (for example 1-4, particularly 1, 2 or 3) heteroatoms selected from nitrogen, oxygen or sulfur. The term heteroaryl includes both monovalent species and divalent species. Examples of heteroaryl groups are monocyclic and bicyclic groups containing from five to twelve ring members, and more usually from five to ten ring members. The heteroaryl group can be, for example, a 5- or 6-membered monocyclic ring or a 9- or 10- membered bicyclic ring, for example a bicyclic structure formed from fused five and six membered rings or two fused six membered rings. Each ring may contain up to about four heteroatoms typically selected from nitrogen, sulfur and oxygen. Typically the heteroaryl ring will contain up to 3 heteroatoms, more usually up to 2, for example a single heteroatom. In one embodiment, the heteroaryl ring contains at least one ring nitrogen atom. The nitrogen atoms in the heteroaryl rings can be basic, as in the case of an imidazole or pyridine, or essentially non- basic as in the case of an indole or pyrrole nitrogen. In general the number of basic nitrogen atoms present in the heteroaryl group, including any amino group substituents of the ring, will be less than five.
[0042] Examples of heteroaryl include furyl, pyrrolyl, thienyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,3,5-triazenyl, benzofuranyl, indolyl, isoindolyl, benzothienyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, benzothiazolyl, indazolyl, purinyl, benzofurazanyl, quinolyl, isoquinolyl, quinazolinyl, quinoxalinyl, cinnolinyl, pteridinyl, naphthyridinyl, carbazolyl, phenazinyl, benzisoquinolinyl, pyridopyrazinyl, thieno[2,3-b]furanyl, 2H-furo[3,2-b]-pyranyl, 5H-pyrido[2,3-d]-o-oxazinyl, lH-pyrazolo[4,3-d]-oxazolyl,
4H-imidazo[4,5-d]thiazolyl, pyrazino[2,3-d]pyridazinyl, imidazo[2,l-b]thiazolyl, imidazo[l,2-b][l,2,4]triazinyl. "Heteroaryl" also covers partially aromatic bi- or polycyclic ring systems wherein at least one ring is an aromatic ring and one or more of the other ring(s) is a non-aromatic, saturated or partially saturated ring, provided at least one ring contains one or more heteroatoms selected from nitrogen, oxygen or sulfur. Examples of partially aromatic heteroaryl groups include for example, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 2-oxo- 1,2,3,4-tetrahydroquinolinyl, dihydrobenzthienyl, dihydrobenzfuranyl, 2,3-dihydro- benzo[l,4]dioxinyl, benzo[l,3]dioxolyl, 2,2-dioxo-l,3-dihydro-2-benzothienyl, 4,5,6,7- tetrahydrobenzofuranyl, indolinyl, l,2,3,4-tetrahydro-l,8-naphthyridinyl, l,2,3,4-tetrahydropyrido[2,3- ?]pyrazinyl and 3,4-dihydro-2H-pyrido[3,2-Z?] [l,4]oxazinyl
[0043] Examples of five membered heteroaryl groups include but are not limited to pyrrolyl, furanyl, thienyl, imidazolyl, furazanyl, oxazolyl, oxadiazolyl, oxatriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl and tetrazolyl groups.
[0044] Examples of six membered heteroaryl groups include but are not limited to pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl and triazinyl.
[0045] A bicyclic heteroaryl group may be, for example, a group selected from:
a) a benzene ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; b) a pyridine ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; c) a pyrimidine ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; d) a pyrrole ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; e) a pyrazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; f) a pyrazine ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; g) an imidazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; h) an oxazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; i) an isoxazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; j) a thiazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; k) an isothiazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; 1) a thiophene ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; m) a furan ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; n) a cyclohexyl ring fused to a 5- or 6-membered heteroaromatic ring containing 1, 2 or 3 ring heteroatoms; and
o) a cyclopentyl ring fused to a 5- or 6-membered heteroaromatic ring containing 1, 2 or 3 ring heteroatoms.
[0046] Particular examples of bicyclic heteroaryl groups containing a six membered ring fused to a five membered ring include but are not limited to benzofuranyl, benzothiophenyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl,
isobenzofuranyl, indolyl, isoindolyl, indolizinyl, indolinyl, isoindolinyl, purinyl (e.g., adeninyl, guaninyl), indazolyl, benzodioxolyl and pyrazolopyridinyl groups.
[0047] Particular examples of bicyclic heteroaryl groups containing two fused six membered rings include, but are not limited to, quinolinyl, isoquinolinyl, chromanyl, thiochromanyl, chromenyl, isochromenyl, chromanyl, isochromanyl, benzodioxanyl, quinolizinyl, benzoxazinyl, benzodiazinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl and pteridinyl groups.
[0048] "Heteroaryl(l-6C)alkyl" means a heteroaryl group covalently attached to a (1- 6C)alkylene group, both of which are defined herein. Examples of heteroaralkyl groups include pyridin-3-ylmethyl, 3-(benzofuran-2-yl)propyl, and the like.
[0049] The term "aryl" means a cyclic or polycyclic aromatic ring having from 5 to 12 carbon atoms. The term aryl includes both monovalent species and divalent species. Examples of aryl groups include, but are not limited to, phenyl, biphenyl, naphthyl and the like. In particular embodiment, an aryl is phenyl.
[0050] The term "aryl(l-6C)alkyl" means an aryl group covalently attached to a (l-6C)alkylene group, both of which are defined herein. Examples of aryl-(l-6C)alkyl groups include benzyl, phenylethyl, and the like
[0051] This specification also makes use of several composite terms to describe groups comprising more than one functionality. Such terms will be understood by a person skilled in the art. For example heterocyclyl(m-nC)alkyl comprises (m-nC)alkyl substituted by
heterocyclyl. [0052] The term "optionally substituted" refers to either groups, structures, or molecules that are substituted and those that are not substituted.
[0053] Where optional substituents are chosen from "one or more" groups it is to be understood that this definition includes all substituents being chosen from one of the specified groups or the substituents being chosen from two or more of the specified groups.
[0054] The phrase "compound of the invention" means those compounds which are disclosed herein, both generically and specifically.
Compounds
[0055] According to a first aspect of the present invention there is provided a compound of formula I:
Figure imgf000010_0001
wherein:
X is O, S, SO or S02;
Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl, (2-6C)alkynyl, or Q is group of the formula:
-L'-Q1
wherein:
L1 is absent or (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene, each of which is optionally substituted with one or more (l-4C)alkyl groups;
Q1 is selected from -OR9, -NR9R10, -S(0)pR9 (wherein p is 0, 1 or 2), -C(0)R9, -C(0)OR9, -OC(0)R9, -C(O)NR9R10, -N(R10)C(O)R9, -N(R10)CON(R10)R9-, -S02N(R9)-, -N(R9)S02-, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
R is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (1- 4C)alkoxy;
Rio is selected from hydrogen or (l-6C)alkyl;
Ri, R2, R3, R4, R5, R6, R7 Re, R21, R22 and R23 are each independently selected from hydrogen, halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,
or a group of the formula:
-L2-L3-R2o
wherein
L2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
L3 is absent or is selected from O, S, SO, S02, OS(0)2, N(Ri3), C(O), CH(ORi3),
C(0)0, OC(O), C(0)N(Ri3), N(Ri3)C(0), N(Ri3)C(0)N(Ri4), S(0)2N(Ri3), or
N(Ri3)S02, wherein R13 and RJ4 are each independently selected from hydrogen or (l-4C)alkyl; and
R20 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R2o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl (l-4C)alkoxy, or a group of the formula:
Figure imgf000011_0001
wherein
L4 is absent or is selected from O, S, SO, S02, N(R25), C(O), CH(OR25), C(0)0, OC(O), C(0)N(R25), N(R25)C(0), N(R25)C(0)N(R26), S(0)2N(R25), or N(R25)S02, wherein R25 and R26 are each independently selected from hydrogen or (l-4C)alkyl; and
R24 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R24 is optionally further substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl or (l-4C)alkoxy;
wherein any nitrogen atoms present in any of the groups Rl5 R2, R3, R4, R5, R6, R7 Rs, R21, R22 and R23 are optionally in the form of an N-oxide;
or a pharmaceutically acceptable salt or solvate thereof.
[0056] According to another aspect of the present invention there is provided a compound of formula la:
Figure imgf000012_0001
wherein:
X is O, S, SO or S02;
Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl or (2-6C)alkynyl, or Q is a group of the formula:
-L'-Q1
wherein:
L1 is absent or (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene, each of which is optionally substituted with one or more (l-4C)alkyl groups; Q1 is selected from -OR9, -NR9R10, -S(0)pR9 (wherein p is 0, 1 or 2), -C(0)R9, -C(0)OR9, -OC(0)R9, -C(0)NR9Rio, -N(Ri0)C(O)R9, -N(R10)CON(R10)R9-, -S02N(R9)-, -N(R9)S02-, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
R9 is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (1- 4C)alkoxy;
Rio is selected from hydrogen or (l-6C)alkyl;
Ri, R2, R3, R4, R5, R6, R7 and Rg are each independently selected from hydrogen, halogeno, trifluoromefhyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,
or a group of the formula:
Figure imgf000013_0001
wherein
L2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
L3 is absent or is selected from O, S, SO, S02, N(Ri3), C(O), CH(ORi3), C(0)0, OC(O), C(0)N(R13), N(Ri3)C(0), N(R13)C(0)N(R14), S(0)2N(R13), or
N(Ri3)S02, wherein Ri3 and R14 are each independently selected from hydrogen or (l-4C)alkyl; and
R20 is (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3-6C)cycloalkyl-(l-
6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R20 is optionally further substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl or (1- 4C)alkoxy;
or a pharmaceutically acceptable salt or solvate thereof.
[0057] Particular novel compounds of the invention include, for example, compounds of the formula I, or pharmaceutically acceptable salts or solvates thereof, wherein, unless otherwise stated, each of X, Q, R1; R2, R3, R4, R5, R6, R7, Rs, R21, R22 and R23 has any of the meanings defined hereinbefore or in any of paragraphs (1) to (41) hereinafter: -
(1) X is O, S or S02;
(2) X is S;
(3) X is O;
(4) X is S02;
(5) Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl or (2-6C)alkynyl, or Q is a group of the formula: wherein:
L1 is (l-6C)alkylene which is optionally substituted with one or more (l-4C)alkyl groups; Q1 is selected from -OR9, -NR9R10, -S(0)pR9 (wherein p is 0, 1 or 2), -C(0)R9,
-C(0)OR9, -OC(0)R9, -C(0)NR9Rio, -N(R10)C(O)R9, -N(Ri0)CON(Ri0)R9-, -S02N(R9)-, -N(R9)S02-, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3- 8C)cycloalkyl, aryl, heterocyclyl or heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
R9 is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
Rio is selected from hydrogen or (l-6C)alkyl;
Q is selected from O , (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl or (2-6C)alkynyl, or Q is a group of the formula:
Figure imgf000014_0001
wherein:
L1 is (l-4C)alkylene which is optionally substituted with one or more (l-2C)alkyl groups; Q1 is selected from -OR9, -NR9Rio, -S(0)pR9 (wherein p is 0, 1 or 2), -C(0)R9,
-C(0)OR9, -OC(0)R9, -C(O)NR9R10, -N(R10)C(O)R9, -N(R10)CON(R10)R9-, -S02N(R9)-, -N(R9)S02-, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3- 6C)cycloalkyl, aryl, heterocyclyl or heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
R9 is selected from hydrogen, (l-4C)alkyl, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-4C)alkyl, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
Rio is selected from hydrogen or (l-2C)alkyl;
Q is selected from 0~ , (l-6C)alkyl, (l-6C)alkoxy, or Q is a group of the formula: wherein:
L1 is (l-2C)alkylene;
Q1 is selected from -OR9, -NR9Ri0, -S(0)pR9 (wherein p is 0, 1 or 2), -C(0)R9,
-C(0)OR9, -OC(0)R9, -C(0)NR9Rio, -N(Ri0)C(O)R9, -N(Ri0)CON(Ri0)R9-, -S02N(R9)-, -N(R9)S02-, (3-6C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3- 6C)cycloalkyl, aryl, heterocyclyl or heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
R9 is selected from hydrogen or (l-4C)alkyl;
selected from hydrogen or (l-2C)alkyl;
selected from O", (l-6C)alkyl, (l-6C)alkoxy, or Q is a group of the formula:
-V-Q
wherein L and Q have any one of the definitions set out herein;
(9) Q is selected from O , (l-6C)alkyl, or (l-6C)alkoxy;
(10) Q is selected from O , (l-4C)alkyl, or (l-4C)alkoxy;
(11) Q is selected from O , (l-2C)alkyl, (l-2C)alkoxy;
(12) Q is O~ or (l-6C)alkyl.
(13) Q is O",or (l-2C)alkyl.
(14) Q is O , or methyl.
(15) Q is methyl.
(16) Q is O".
(17) Ri, R2, R3, R4, R5, R6, R7 and Rs are each independently selected from hydrogen,
halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, or a group of the formula:
-L2-L3-R20
wherein
L2 is absent or (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
L3 is absent or is selected from O, S, SO, S02, N(Ri3), C(O), CH(ORi3), C(0)0,
OC(O), C(0)N(R13), N(Ri3)C(0), N(R13)C(0)N(R14), S(0)2N(R13), or
N(Ri3)S02, wherein Ri3 and R14 are each independently selected from hydrogen or (l-2C)alkyl; and
R20 is (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3-6C)cycloalkyl-(l- 6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l- 6C)alkyl and wherein R20 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
and R2i, R22 and R23 are hydrogen; (18) Ri, R2, R3, R4, R5, R6, 7 and R8 are each independently selected from hydrogen, halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, or a group of the formula:
-L2-L3-R2o
wherein
L2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
L3 is absent or is selected from O, S, SO, S02, N(Ri3), C(O), CH(ORi3), C(0)N(Ri3), N(Ri3)C(0), S(0)2N(Ri3), or N(Ri3)S02, wherein Ri3 and Ri4 are each independently selected from hydrogen or (l-2C)alkyl; and
R20 is (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3-6C)cycloalkyl-(l- 6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l- 6C)alkyl and wherein R20 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
and R2i, R22 and R23 are hydrogen;
(19) Ri, R2, R3, R4, R5, R6, R7 and Rg are each independently selected from hydrogen,
halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, carbamoyl, (1- 6C)alkyl, or a group of the formula:
-L2-L3-R2o
wherein
L2 is absent or a methylene or a -CH=CH- linker;
L3 is absent or is selected from O, S, SO, S02, N(Ri3), C(O), CH(ORi3), C(0)N(Ri3), N(Ri3)C(0), N(R13)C(0)N(R14), S(0)2N(Ri3), or N(Ri3)S02, wherein Ri3 and R14 are each independently selected from hydrogen or (1- 2C)alkyl; and
R20 is (l-6C)alkyl or (3-6C)cycloalkyl, and wherein R2o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-2C)alkoxy;
and R2i, R22 and R23 are hydrogen;
(20) Ri, R2, R3, R4, R5, R6, R7 and R8 are each independently selected from hydrogen,
halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, carbamoyl, (1- 4C)alkyl, or a group of the formula:
-L2-L -R2o
wherein L2 is absent;
L3 is absent or is selected from O, S, SO, S02, N(Ri3), C(O), CH(ORi3), C(0)N(Ri3), N(Ri3)C(0), S(0)2N(Ri3), or N(Ri3)S02, wherein Ri3 and Ri4 are each independently selected from hydrogen or (l-2C)alkyl; and
R20 is (l-4C)alkyl which is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino or (l-2C)alkoxy;
and R21, R22 and R23 are hydrogen;
Ri, R2, R3, R4, R5, R6, R7 Re, R21, R22 and R23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R
wherein
L2 is absent or a (l-6C)alkylene or (2-6C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, C(0)N(Ri3),
N(Ri3)C(0), wherein Ri3 is selected from hydrogen or (l-4C)alkyl; and
R20 is hydrogen, (l-6C)alkyl, heteroaryl, or heterocyclyl and wherein R20 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-4C)alkoxy, or a group of the formula:
L4-R24
wherein
L4 is N(R25), wherein R25 is independently selected from hydrogen or (1- 4C)alkyl; and
R24 is (l-6C)alkyl;
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R?,Rs R21, R22 and R23 are optionally in the form of a N-oxide;
Ri, R2, R3, R4, R5, R6, 7 s, R21, R22 and R23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R;
wherein
L2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, C(0)N(Ri3),
N(Ri3)C(0), wherein Ri3 is selected from hydrogen or (l-2C)alkyl; and R20 is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R2o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy, or a group of the formula:
L -R24
wherein
L is N(R25), wherein R25 is (l-2C)alkyl; and
R24 is (l-2C)alkyl;
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7, Re, R21, R22 and R23 are optionally in the form of a N-oxide;
(23) Ri, R2, R3, R4, R5, R6, R7 Rs, R21, R22 and R23 are each independently selected from hydrogen, fluoro, hydroxy, or a group of the formula:
-L2-L3-R;
wherein
L2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(R13), C(0)0, C(0)N(Ri3), N(Ri3)C(0), wherein R13 is selected from hydrogen or (l-2C)alkyl; and R20 is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R20 is optionally further substituted by one or more substituents independently selected from fluoro, hydroxy, (l-2C)alkoxy, or a group of the formula:
L4-R24
wherein
L4 is N(R25), wherein R25 is (l-2C)alkyl; and
R24 is (l-2C)alkyl;
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7, Re, R21, R22 and R23 are optionally in the form of a N-oxide;
Ri, R2, R3, R4, R5, R6, R7 R8, R21, R22 and R23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R2o
wherein
L2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, N(Ri3)C(0), wherein Ri3 is selected from hydrogen or (l-2C)alkyl; and R20 is (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R2o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy, or a group of the formula:
L -R24
wherein
L is N(R25), wherein R25 is (l-2C)alkyl; and
R24 is (l-2C)alkyl;
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7, Re, R21, R22 and R23 are optionally in the form of a N-oxide;
(25) Ri, R2, R3, R4, R5, R6, 7 s, R21, R22 and R23 are each independently selected from
hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R;
wherein
L2 is absent or a (l-3C)alkylene or (2-3C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, N(Ri3)C(0), wherein Ri3 is selected from hydrogen or (l-2C)alkyl; and
R20 is (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R20 is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy,
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R5, R7,
Rg, R21, R22 and R23 are optionally in the form of a N-oxide;
Ri, R2, R3, R4, R5, R6, 7 s, R21, R22 and R23 are each independently selected from hydrogen, halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,
or a group of the formula:
-L2-L3-R
wherein
L2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
L3 is absent or is selected from O, S, SO, S02, OS(0)2, N(Ri3), C(O), CH(ORi3), C(0)0, OC(O), C(0)N(R13), N(R13)C(0), N(R13)C(0)N(R14), S(0)2N(R13), or
N(Ri3)S02, wherein Ri3 and RJ4 are each independently selected from hydrogen or (l-4C)alkyl; and
R20 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R2o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl (l-4C)alkoxy
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R5, R7, Rg, R21, R22 and R23 are optionally in the form of a N-oxide;
Ri, R2, R3, R4, R5, R6, 7 s, R2i, 22 and R23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R2o
wherein
L2 is absent or a (l-6C)alkylene, or (2-6C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, C(0)N(Ri3),
N(Ri3)C(0), wherein Ri3 is selected from hydrogen or (l-4C)alkyl; and
R20 is hydrogen, (l-6C)alkyl, heteroaryl, or heterocyclyl and wherein R2o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-4C)alkoxy
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7,
Rg, R2i, R22 and R23 are optionally in the form of a N-oxide;
Ri, R2, R3, R4, R5, R6, 7 s, 2i, R22 and R23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R2o
wherein
L2 is absent or a (l-3C)alkylene, or (2-3C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, C(0)N(Ri3),
N(Ri3)C(0), wherein R13 is selected from hydrogen or (l-2C)alkyl; and
R2o is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R2o is optionally further substituted by one or more substituents independently selected from oxo, halogeno, hydroxy, (l-2C)alkoxy;
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7,
Rg, R21, R22 and R23 are optionally in the form of a N-oxide;
Ri, R2, R3, R4, R5, R6, 7 s, 2i> R22 and R23 are each independently selected from hydrogen, fluoro, hydroxy, or a group of the formula:
-L2-L3-R2o
wherein L2 is absent or a (l-3C)alkylene, or (2-3C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, C(0)N(Ri3),
N(Ri3)C(0), wherein R13 is selected from hydrogen or methyl; and
R20 is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R20 is optionally further substituted by one or more substituents independently selected from oxo, fluoro, hydroxy, or methoxy;
and wherein any nitrogen atoms present in any of the groups Rl5 R2, R3, R4, R5, R , R7 Rs, R21, R22 and R23 are optionally in the form of a N-oxide;
(30) Ri and R2 are each independently a substituent group as defined hereinbefore (in
particular a substituent group as defined in any one of paragraphs (17) to (29) above); and
R3, R4, R5, R6, R7, Re, R21, R22 and R23 are each independently selected from hydrogen, fluoro, CF3, OCF3 or (l-2C)alkoxy.
(31) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and R3, R4, R5, R5, R7 R8, R2i, R22 and R23 are each independently selected from hydrogen, fluoro, CF3, OCF3 or (l-2C)alkoxy.
(32) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto four of R3, R4, R5, R6, R7 R8, R2i, R22 and R23 are each independently selected from fluoro, CF3, OCF3 or (l-2C)alkoxy and the others are hydrogen.
(33) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto three of R3, R4, R5, R6, R7 Rs, R21, R22 and R23 are each independently selected from fluoro, CF3, OCF3 or (l-2C)alkoxy and the others are hydrogen.
(34) Ri and R2 are each independently a substituent group as defined hereinbefore (in
particular a substituent group as defined in any one of paragraphs (17) to (29) above); and R3, R4, R5, R6, R7 Re, R21, R22 and R23 are each independently selected from hydrogen, fluoro, CF3 or OCF3.
(35) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and R3, R4, R5, R6, R7 Rs, R21, R22 and R23 are each independently selected from hydrogen, fluoro, CF3 or OCF3. (36) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto four of R3, R4, R5, R6, R7 R8, R2i, R22 and R23 are each independently selected from fluoro, CF3 or OCF3 and the others are hydrogen.
(37) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto three of R3, R4, R5, R6, R7 R8, R2i, R22 and R23 are each independently selected from fluoro, CF3 or OCF3 and the others are hydrogen.
(38) Ri and R2 are each independently a substituent group as defined hereinbefore (in
particular a substituent group as defined in any one of paragraphs (17) to (29) above); and R3, R4, R5, R6, R7 Rs, R21, R22 and R23 are each independently selected from hydrogen, fluoro, CF3 or OCF3.
(39) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and R3, R4, R5, R5, R7 Rs, R21, R22 and R23 are each independently selected from hydrogen or fluoro.
(40) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto four of R3, R4, R5, R6, R7 R8, R2i, R22 and R23 are fluoro and the others are hydrogen.
(41) one of Ri and R2 is a substituent group other than hydrogen as defined hereinbefore (in particular a substituent group as defined in any one of paragraphs (17) to (29) above) and the other is hydrogen; and upto three of R3, R4, R5, R6, R7 Rs, R21, R22 and R23 are fluoro and the others are hydrogen.
[0058] Suitably, X is as defined in any one of paragraphs 1 to 4 above. In a particular group of compounds, X is O or S.
[0059] Suitably, Q is as defined in any one of paragraphs 5 to 16 above. In a particular group of compounds, Q is O" or methyl.
[0060] Suitably, Ri, R2, R3, R4, R5, R6, R7 Rs, R21, R22 and R23 have any one of the definitions set out in paragraphs 17 to 41 above. In a particular group of compounds, Rl5 R2, R3, R4, R5, R6, R7 R8, R2i, R22 and R23 have any one of the definitions set out in paragraphs 29 to 41 above. In a further group of compounds, Ri, R2, R3, R^, R5, R6, R7 Rs, R2i, R22 and R23 have any one of the definitions set out in paragraphs 30 to 41 above. [0061] In a particular group of compounds of the invention, up to four of Rl 5 R2, R3, R4, R5, R6, R7, Re, R21, R22 and R23 are a substituent group other than hydrogen.
[0062] In a further group of compounds of the invention, up to three of Ri, R2, R3, R4, R5, R6, R7, Re, R21, R22 and R23 are a substituent group other than hydrogen.
[0063] In a further group of compounds of the invention, one or two of Ri, R2, R3, R4, R5, R6, R7, Rg, R21, R22 and R23 is/are a substituent group other than hydrogen.
[0064] In a further group of compounds of the invention, one of Rl 5 R2, R3, R4, R5, R6, R7, Rs, 215 R22 and R23 is a substituent group other than hydrogen.
[0065] In a particular group of compounds of the invention, Rg, R21, R22 and R23 are hydrogen, i.e. the compounds have the structural formula II shown below:
Figure imgf000023_0001
wherein X, Q, Ri, R2, R3, R4, R5, R6 and R7 have any one of the meanings defined hereinbefore, or a pharmaceutically acceptable salt or solvate thereof.
[0066] In a further group of compounds of the invention, the groups R4, R5, R6, R7, Rs, R21, R22 and R23 are all hydrogen, i.e. the compounds have the structural formula III shown below:
Figure imgf000023_0002
III
wherein X, Q, Rl5 R2 and R3 have any one of the meanings defined hereinbefore, or a pharmaceutically acceptable salt or solvate thereof. [0067] In yet a further group of compounds of the invention, the groups R3, R4, R5, R6, R7, R8, R21, R22 and R23 are all hydrogen, i.e. the compounds have the structural formula IV shown below:
Figure imgf000024_0001
IV
wherein X, Q, Ri and R2 have any one of the meanings defined hereinbefore; or a
pharmaceutically acceptable salt or solvate thereof.
[0068] In a particular group of compounds of formula IV, one of Ri or R2 is hydrogen.
[0069] In yet a further group of compounds of the invention,
X is as defined herein before (and is particularly as described in any one of paragarphs (1) to (4) above);
Q is O" or (l-6C)alkyl (and is particularly O" or methyl);
Ri, R2 and R3 each independently have any one of the definitions set out hereinbefore (and in particular are as defined in any one of paragraphs (17) to (41) above); and
R4, R5, R6, R7, Re, R21, R22 and R23 each have any one of the definitions set out hereinbefore;
or a pharmaceutically acceptable salt or solvate thereof.
[0070] In yet a further group of compounds of the invention,
X is O, S or S02;
Q is O" or methyl;
and Ri, R2, R3, R4, R5, R6, R7, Rs, R21, R22 and R23 each have any one of the definitions set out in paragraphs (30) to (41) above;
or a pharmaceutically acceptable salt or solvate thereof.
[0071] The various functional groups and substituents making up the compounds of the invention are typically chosen such that the molecular weight of the compound does not exceed 1000. More usually, the molecular weight of the compound will be less than 750, for example less than 700, or less than 650, or less than 600, or less than 550. More preferably, the molecular weight is less than 525 and, for example, is 500 or less.
[0072] Particular compounds of the invention include any one of the following:
13 -methylthiochromeno [4, 3 ,2-gh] phenanfhridin- 13 -ium;
13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium; or
13 -methylthiochromeno [4,3 ,2-gh] phenanthridin- 13 -ium- 10,10-dioxide ;
2-(furan-2-yl)-13-methylchromeno[4,3,2-g¾phenanthridin-13-ium;
2-(furan-3-yl)-13-methylchromeno[4,3,2- /z]phenanthridin-13-ium;
2-hydroxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium;
2-(2-hydroxyethoxy)- 13-methylchromeno[4,3,2-g¾]phenanfhridin- 13-ium;
2-acetamido- 13-methylchromeno [4,3 ,2-g i]phenanthridin- 13-ium;
13-methyl-2-(N-methylacetamido)chromeno[4,3,2-g/z]phenanthridin-13-ium;
2-amino-13-methylthiochromeno[4,3,2-gA]phenanthridin-13-ium;
13-methyl-2-(lH-pyrazol-4-yl)thiochromeno[4,3,2- z]phenanthridin-13-ium;
2-(3-acetamidoprop- 1 -ynyl)- 13 -methylthiochromeno [4, 3, 2-g¾ phenanthridin- 13-ium;
13-methyl-2-(thiophen-3-yl)thiochromeno[4,3,2-gA]phenanthridin-13-ium;
6, 10-dimethoxy- 13-methylchromeno[4,3 ,2-gh] phenanthridin- 13-ium;
6, 10-dimethoxy- 13-methylchromeno[4,3 ,2-gh] phenanthridin- 13-ium;
6.10- bis(2-hydroxyethoxy)-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium;
1 ,3-difluoro-6, 10-dimethoxy- 13-methylchromeno [4,3 ,2-g z]phenanthridin- 13-ium;
1 , 1 l-difluoro-2-methoxy- 13-methylchromeno[4,3 ,2-gh] phenanthridin- 13-ium;
1.11- difluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2-g¾]phenanthridin-13-ium;
3 , 9-dimethoxy- 13 -methylchromeno [4, 3 ,2-gh] phenanthridin- 13 -ium;
2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
13-methyl-2-(thiophen-3-yl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
2-(3-(dimethylamino)prop- 1 -ynyl)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
13 -methyl-2-( 1 H-pyrazol-4-yl)chromeno [4,3 ,2-gh] phenanthridin- 13-ium;
2-methoxy- 13-methylchromeno [4,3 ,2-g/z]phenanthridin- 13-ium;
2-(methoxymethyl)-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium;
2-((dimethylamino)methyl)-13-methylchromeno[4,3,2-gA]phenanthridin-13-ium;
2-methoxy- 13-methylthiochromeno[4,3,2-g z]phenanthridin-13-ium;
2-hydroxy- 13-methylthiochromeno [4,3 ,2-g z]phenanthridin- 13-ium;
4, 11 -difluoro-2-methoxy- 13-methylchromeno [4,3 ,2-gh] phenanthridin- 13-ium;
2-(3-acetamidoprop- 1 -ynyl)- 13-methylchromeno [4, 3, 2-g/z] phenanthridin- 13-ium; 2- (dimethylamino)-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
13-methyl-2-morpholinochromeno[4,3,2- z]phenanthridin-13-ium;
ll-fluoro-3-methoxy-13-methylchromeno[4,3,2-^]phenanthridin-13-ium;
ll-fluoro-13-methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2-gA]phenanthridin-13-ium; 13-methyl-2-(pyridin-4-yl)chromeno[4,3,2- z]phenanthridin-13-ium;
4-(13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium-2-yl)pyridine 1-oxide;
3,11 -difluoro- 13-methylchromeno [4,3,2-gA]phenanthridin- 13-ium;
1 , 12-difluoro- 13-methylchromeno [4,3,2-g/z]phenanthridin- 13-ium;
1- fluoro-2-methoxy-13-methyl-10-(trifluoromethoxy)chromeno[4,3,2-g z]phenanthridin-13-ium; 3-methoxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
13 -methyl-3 -(trif uoromethylsulfonyloxy)chromeno [4, 3 ,2-gh] phenanthridin- 13-ium;
13 -methyl-3 -( 1 H-pyrazol-4-yl)chromeno [4,3 ,2-gh] phenanthridin- 13 -ium;
13-methyl-3-(thiophen-3 -yl)chromeno [4,3 ,2-g/i]phenanthridin- 13-ium;
3- (3-(dimethylamino)prop- 1 -ynyl)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
3-(3-acetamidoprop-l-ynyl)-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium;
3, 11-dimethoxy- 13-methylchromeno[4,3,2-g¾phenanthridin- 13-ium;
3,11 -difluoro- 13 -methyl-2-( 1 H-pyrazol-4- yl)chromeno [4,3 ,2-g i]phenanthridin- 13 -ium;
3- fluoro-2-(methoxycarbonyl)- 13 -methylchromeno [4,3 ,2-g/i] phenanthridin- 13-ium;
3,11 -difluoro-2-(methoxycarbonyl)- 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13-ium;
2-carbamoyl- 1,11 -difluoro- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
2- carboxy- 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13 -ium;
3, 1 l-difluoro-2-methoxy- 13-methylchromeno[4,3 ,2-gh] phenanthridin- 13-ium;
2-(ethoxycarbonyl)- 13 -methylchromeno [4,3,2-g/z]phenanthridin- 13-ium;
2-(isopropoxycarbonyl)-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium;
2-carbamoyl- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
2-carbamoyl-3-fluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
4,11 -difluoro-2-(methoxycarbonyl)- 13 -methylchromeno [4,3 ,2-g/i] phenanthridin- 13-ium;
2-carbamoyl-3,l l-difluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
13-methyl-2-(morpholinomethyl)chromeno[4,3,2-g i]phenanthridin-13-ium;
1,11 -difluoro-2-(2-hydroxyethoxy) - 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13 -ium;
4- ((13-methylchromeno[4,3,2-g¾phenanthridin-13-ium-2-yl)methyl)morpholine 4-oxide;
2-(2-amino-2-oxoethoxy)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
l,ll-difluoro-2-hydroxy-13-methylchromeno[4,3,2-gA]phenanthridin-13-ium;
3,ll-difluoro-2-(isopropoxycarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium; 2-(methoxymethoxy)-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
2-((2-hydroxyethoxy)carbonyl)- 13 -methylchromeno [4,3 ,2-g ?]phenanthridin- 13-ium;
11 -fluoro-2-(methoxycarbonyl)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
2-carbamoyl-4, 11 -difluoro- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
11 -fluoro-2-(methoxymethyl)- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
4,l l-difluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
4,l l-difluoro-13-methyl-2-((tetrahydrofuran-2-yloxy)carbonyl)chromeno[4,3,2- g j]phenanthridin- 13-ium;
4, 1 l-difluoro-2-((3-hydroxypropoxy)carbonyl)- 13-methylchromeno[4,3,2-g¾]phenanthridin- 13- ium;
4,l l-difluoro-2-((2-methoxyethoxy)carbonyl)-13-methylchromeno[4,3,2-g/z]phenanthridin-13- ium;
11 -fluoro- 13-methyl-2-(morpholinomethyl)chromeno[4,3,2-g z]phenanthridin- 13-ium;
l l-fluoro-13-methyl-2-(3-methyl-l,2,4-oxadiazol-5-yl)chromeno[4,3,2-g z]phenanthridin-13- ium;
2-((dimethylamino)methyl)- 11 -fluoro- 13-methylchromeno[4,3,2-g¾phenanthridin- 13-ium;
2-carboxy-l l-fluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium;
2-(2-amino-2-oxoethoxy)-4, 11 -difluoro- 13-methylchromeno[4,3,2-g i]phenanthridin- 13-ium;
2-carbamoyl- 11 -fluoro- 13-methylchromeno[4,3,2-g¾phenanthridin- 13-ium;
l,l l-difluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3,2-g¾]phenanthridin-13-ium;
4,l l-difluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
4,l l-difluoro-13-methyl-2-(pyrimidin-5-yl)chromeno[4,3,2-g/z]phenanthridin-13-ium;
4,l l-difluoro-2-(2-methoxypyridin-3-yl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium; l l-fluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
l l-fluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3,2-g/j]phenanthridin-13-ium;
11 -fluoro- 13-methyl-2-(pyrimidin-5-yl)chromeno[4,3,2-g z]phenanthridin- 13-ium;
11 -fluoro-2-(2-methoxypyridin-3-yl)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
4,11 -difluoro-2-(6-methoxypyridin-2-yl)- 13-methylchromeno [4,3 ,2-g z]phenanthridin- 13-ium;
4,l l-difluoro-13-methyl-2-(2-oxo-l,2-dihydropyridin-3-yl)chromeno[4,3,2-g i]phenanthridin- 13-ium;
11 -fluoro-2-(6-methoxypyridin-2-yl)- 13-methylchromeno[4,3,2-g¾]phenanthridin- 13-ium; l l-fluoro-13-methyl-2-((oxetan-3-yloxy)carbonyl)chromeno[4,3,2-g z]phenanthridin- 13-ium; l l-fluoro-13-methyl-2-(3-methyl-l,2,4-oxadiazol-5-yl)chromeno[4,3,2-g/z]phenanthridin-13- ium; 2-((l-(tert-butoxycarbonyl)azetidin-3-yloxy)carbonyl)-l l-fluoro-13-methylchromeno[4,3,2- g/z]phenanthridin-13-ium;
11 -lluoro- 13-methyl-2-(2-oxo-l ,2-dihydropyridin-3-yl)chromeno[4,3,2-g z]phenanthridin- 13- ium;
11 -fluoro-13-methyl-2-(2-oxo- 1 ,2-dihydropyridin-3-yl)chromeno[4,3,2-g z]phenanthridin- 13- ium;
13-methyl-2-(pyrimidin-2-yloxy)chromeno[4,3,2-gA]phenanthridin-13-ium;
l l-fluoro-13-methyl-2-(5-methyl-13,4-oxadiazol-2-yl)chromeno[4,3,2-g i]phenanthridin-13- ium;
13-methyl-2-(lH-pyrazol-4-yl)-l l-(trifluoromethox
2-(methoxycarbonyl)- 13-methyl- 11 -(trM^
ium;
in association with a pharmaceutically acceptable anion.
[0073] Further compounds of the invention include any one of the following:
N,N-dimethyl-l-(13-methylchromeno[4,3,2-g z]phenanthridin-13-ium-2-yl)methanamine oxide; 2-((2-(dimethylamino)ethoxy)carbonyl)-l l-fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin- 13-ium;
2-(l l-fiuoro-13-methylchromeno[4,3,2-g/i]phenanthridin-2— 13-iumcarbonyloxy)-N,N- dimethylethanamine oxide;
2-(3-(dimethylamino)prop-l-ynyl)-l l-fluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13- ium;
1- (l l-fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium-2-yl)-N,N- dimethylmethanamine oxide;
4-((l l-fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium-2-yl)methyl)morpholine 4- oxide;
l l-fluoro-2-(2-hydroxyethoxy)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
11 -fluoro-3-(methoxycarbonyl)- 13-methylchromeno[4,3,2-g i]phenanthridin- 13-ium;
l l-fiuoro-3-(2-hydroxyethoxy)-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium;
2- (3-(dimethylamino)prop-l-ynyl)-l,l l-difluoro-13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
2-((dimethylamino)methyl)-l,l l-difluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium; 1 -( 1 , 11 -difluoro- 13 -methylchromeno[4,3,2-g j]phenanthridin- 13-ium-2-yl)-N,N- dimethylmethanamine oxide;
1,11 -difluoro- 13-methyl-2-(morpholinomethyl)chromeno [4,3 ,2-gA]phenanthridin- 13-ium; 4-((l,l l-difluoro-13-methylchrom
4-oxide;
2-(ethoxycarbonyl)-3 , 11 -difluoro- 13-methylchromeno [4,3 ,2-g i]phenanthridin- 13-ium;
2-(tert-butoxycarbonyl)-3, 11 -difluoro- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium; 3,l l-difluoro-13-methyl-2-((tetrahydrofuran-3-yloxy)carbonyl)chromeno[4,3,2- gA]phenanthridin-13-ium;
6, 10-difluoro-2-(methoxycarbonyl)- 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13-ium;
6, 10-difluoro-3-(methoxycarbonyl)- 13 -methylchromeno [4,3 ,2-g/i]phenanfhridin- 13-ium;
in association with a pharmaceutically acceptable anion.
[0074] Suitable pharmaceutically acceptable anions include, for example,
trifluoromethanesulfonate, methanesulfonate, iodide, chloride, fluorise, acetate anions.
[0075] The compounds of the invention may be present in any pharmaceutically acceptable salt form. Typically, as the compounds of the invention are positively charged, they are suitably associated with one or more pharmaceutically acceptable anions. The compound of the invention will suitably possess a +1 charge, although other charged substituent groups could be present. The associated anion may carry a -1 charge, in which case the molar ratio of anionrcation is 1:1, or , alternatively, the anion may carry a -2 or -3 charge, in which case the molar ratio of anion ation is will be 1:2 or 1:3, respectively.
[0076] In one embodiment, the anion is independently derived from one or more of the following acids: hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, phosphorous acetic, propionic, succinic, gycolic, stearic, lactic, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetyoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, trifluoromethanesulfonic, ethanesulfonic, ethane disulfonic, oxalic, isethionic, and valeric.
[0077] Compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed "isomers". Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers".
Stereoisomers that are not mirror images of one another are termed "diastereomers" and those that are non-superimposable mirror images of each other are termed "enantiomers". When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (-)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a "racemic mixture".
[0078] The compounds of this invention may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see discussion in Chapter 4 of "Advanced Organic Chemistry", 4th edition J. March, John Wiley and Sons, New York, 2001), for example by synthesis from optically active starting materials or by resolution of a racemic form. Some of the compounds of the invention may have geometric isomeric centres (E- and Z- isomers). It is to be understood that the present invention encompasses all optical, diastereoisomers and geometric isomers and mixtures thereof that possess telomerase inhibitory activity.
[0079] The present invention also encompasses compounds of the invention as defined herein which comprise one or more isotopic substitutions. For example, H may be in any isotopic form, including ¾, 2H(D), and 3H (T); C may be in any isotopic form, including 12C, 13C, and 14C; and O may be in any isotopic form, including 160 and180; and the like.
[0080] It is also to be understood that certain compounds of the invention may exist in solvated as well as unsolvated forms such as, for example, hydrated forms. It is to be understood that the invention encompasses all such solvated forms that possess telomerase inhibitory activity.
[0081] It is also to be understood that certain compounds of the invention may exhibit polymorphism, and that the invention encompasses all such forms that possess telomerase inhibitory activity.
[0082] Compounds of the invention may exist in a number of different tautomeric forms and references to compounds of the formula I include all such forms. For the avoidance of doubt, where a compound can exist in one of several tautomeric forms, and only one is specifically described or shown, all others are nevertheless embraced by the structural formulae presented herein. Examples of tautomeric forms include keto-, enol-, and enolate-forms, as in, for example, the following tautomeric pairs: keto/enol (illustrated below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime, thioketone/enethiol, and nitro/aci-nitro.
Figure imgf000030_0001
keto enol enolate [0083] Compounds of the invention containing an amine function may also form N-oxides. A reference herein to a compound of the invention that contains an amine function also includes the N-oxide. Where a compound contains several amine functions, one or more than one nitrogen atom may be oxidised to form an N-oxide. Particular examples of N-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle. N-Oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a per-acid (e.g. a peroxycarboxylic acid), see for example Advanced Organic Chemistry, by Jerry March, 4th Edition, Wiley Interscience, pages. More particularly, N-oxides can be made by the procedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514) in which the amine compound is reacted with m-chloroperoxybenzoic acid (MCPBA), for example, in an inert solvent such as dichloromethane.
[0084] The compounds of the invention may be administered in the form of a pro-drug which is broken down in the human or animal body to release a compound of the invention. A pro-drug may be used to alter the physical properties and/or the pharmacokinetic properties of a compound of the invention. A pro-drug can be formed when the compound of the invention contains a suitable group or substituent to which a property-modifying group can be attached. Examples of pro-drugs include in vivo cleavable ester derivatives that may be formed at a carboxy group or a hydroxy group in a compound of the invention and in-vivo cleavable amide derivatives that may be formed at a carboxy group or an amino group in a compound of the invention.
[0085] Accordingly, the present invention includes those compounds of the invention as defined hereinbefore when made available by organic synthesis and when made available within the human or animal body by way of cleavage of a pro-drug thereof. Accordingly, the present invention includes those compounds of the invention that are produced by organic synthetic means and also such compounds that are produced in the human or animal body by way of metabolism of a precursor compound, that is a compound of the invention may be a
synthetically-produced compound or a metabolically-produced compound.
[0086] A suitable pharmaceutically acceptable pro-drug of a compound of the invention is one that is based on reasonable medical judgement as being suitable for administration to the human or animal body without undesirable pharmacological activities and without undue toxicity.
[0087] Various forms of pro-drug have been described, for example in the following documents :- a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985); b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier, 1985);
c) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and
H. Bundgaard, Chapter 5 "Design and Application of Pro-drugs", by H. Bundgaard p. 113-191 (1991);
d) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);
e) H. Bundgaard, et ah, Journal of Pharmaceutical Sciences, 77, 285 (1988);
f) N. Kakeya, et al, Chem. Pharm. Bull., 32, 692 (1984);
g) T. Higuchi and V. Stella, "Pro-Drugs as Novel Delivery Systems", A.C.S. Symposium Series, Volume 14; and
h) E. Roche (editor), "Bioreversible Carriers in Drug Design", Pergamon Press, 1987.
[0088] A suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a carboxy group is, for example, an in vivo cleavable ester thereof. An in vivo cleavable ester of a compound of the invention containing a carboxy group is, for example, a pharmaceutically acceptable ester which is cleaved in the human or animal body to produce the parent acid. Suitable pharmaceutically acceptable esters for carboxy include
Ci_6alkyl esters such as methyl, ethyl and ieri-butyl, Ci-6alkoxymethyl esters such as
methoxymethyl esters, Ci-6alkanoyloxymethyl esters such as pivaloyloxymethyl esters,
3-phthalidyl esters, C3_gcycloalkylcarbonyloxy- Ci_6alkyl esters such as
cyclopentylcarbonyloxymethyl and 1-cyclohexylcarbonyloxyethyl esters,
2-oxo-l,3-dioxolenylmethyl esters such as 5-methyl-2-oxo-l,3-dioxolen-4-ylmethyl esters and Ci ealkoxycarbonyloxy- Ci^alkyl esters such as methoxycarbonyloxymethyl and
1 -methoxycarbonyloxyethyl esters .
[0089] A suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a hydroxy group is, for example, an in vivo cleavable ester or ether thereof. An in vivo cleavable ester or ether of a compound of the invention containing a hydroxy group is, for example, a pharmaceutically acceptable ester or ether which is cleaved in the human or animal body to produce the parent hydroxy compound. Suitable pharmaceutically acceptable ester forming groups for a hydroxy group include inorganic esters such as phosphate esters (including phosphoramidic cyclic esters). Further suitable pharmaceutically acceptable ester forming groups for a hydroxy group include Ci ioalkanoyl groups such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups, Ci_ioalkoxycarbonyl groups such as ethoxycarbonyl, N,N -(Ci-6)2carbamoyl, 2-dialkylaminoacetyl and 2-carboxyacetyl groups. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N- alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin-l-ylmethyl and 4-(C1- 4alkyl)piperazin-l-ylmethyl. Suitable pharmaceutically acceptable ether forming groups for a hydroxy group include a-acyloxyalkyl groups such as acetoxymethyl and pivaloyloxymethyl groups.
[0090] A suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a carboxy group is, for example, an in vivo cleavable amide thereof, for example an amide formed with an amine such as ammonia, a Ci-4alkylamine such as methylamine, a (Ci_ 4alkyl)2amine such as dimethylamine, N-ethyl-N-methylamine or diethylamine, a Ci_4alkoxy- C2- 4alkylamine such as 2-methoxyethylamine, a phenyl-Ci_4alkylamine such as benzylamine and amino acids such as glycine or an ester thereof.
[0091] A suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses an amino group is, for example, an in vivo cleavable amide derivative thereof.
Suitable pharmaceutically acceptable amides from an amino group include, for example an amide formed with Ci-ioalkanoyl groups such as an acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl,
N,N-dialkylaminomethyl, morpholinomethyl, piperazin-l-ylmethyl and
4-(Ci_4alkyl)piperazin- 1 -ylmethyl.
[0092] The in vivo effects of a compound of the invention may be exerted in part by one or more metabolites that are formed within the human or animal body after administration of a compound of the invention. As stated herein before, the in vivo effects of a compound of the invention may also be exerted by way of metabolism of a precursor compound (a pro-drug).
[0093] It shall also be appreciated that compounds of invention may also be covalently linked (at any suitable position) to other groups such as, for example, solubilising moieties (for example, PEG polymers), moieties that enable them to be bound to a solid support (such as, for example, biotin-containing moieties), and targeting ligands (such as antibodies or antibody fragments).
Synthesis
[0094] The compounds of the present invention can be prepared in accordance with the general process described below and/or by the processes described in the accompanying examples.
[0095] In the description of the synthetic methods described below and in the referenced synthetic methods that are used to prepare the staring materials, it is to be understood that all proposed reaction conditions, including choice of solvent, reaction atmosphere, reaction temperature, duration of the experiment and workup procedures, can be selected by a person skilled in the art. [0096] It is understood by one skilled in the art of organic synthesis that the functionality present on various portions of the molecule must be compatible with the reagents and reaction conditions utilised.
[0097] It will be appreciated that during the synthesis of the compounds of the invention in the processes defined below, or during the synthesis of certain starting materials, it may be desirable to protect certain substituent groups to prevent their undesired reaction. The skilled chemist will appreciate when such protection is required, and how such protecting groups may be put in place, and later removed.
[0098] For examples of protecting groups see one of the many general texts on the subject, for example, 'Protective Groups in Organic Synthesis' by Theodora Green (publisher: John Wiley & Sons). Protecting groups may be removed by any convenient method described in the literature or known to the skilled chemist as appropriate for the removal of the protecting group in question, such methods being chosen so as to effect removal of the protecting group with the minimum disturbance of groups elsewhere in the molecule.
[0099] Thus, if reactants include, for example, groups such as amino, carboxy or hydroxy it may be desirable to protect the group in some of the reactions mentioned herein.
[00100] By way of example, a suitable protecting group for an amino or alkylamino group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or i-butoxycarbonyl group, an
arylmethoxycarbonyl group, for example benzyloxycarbonyl, or an aroyl group, for example benzoyl. The deprotection conditions for the above protecting groups necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or
alkoxycarbonyl group or an aroyl group may be removed by, for example, hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide.
Alternatively an acyl group such as a teri-butoxycarbonyl group may be removed, for example, by treatment with a suitable acid as hydrochloric, sulfuric or phosphoric acid or trifluoroacetic acid and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon, or by treatment with a Lewis acid for example boron tris(trifluoroacetate). A suitable alternative protecting group for a primary amino group is, for example, a phthaloyl group which may be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or with hydrazine.
[00101] A suitable protecting group for a hydroxy group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an aroyl group, for example benzoyl, or an arylmethyl group, for example benzyl. The deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or an aroyl group may be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium, sodium hydroxide or ammonia. Alternatively an arylmethyl group such as a benzyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.
[00102] A suitable protecting group for a carboxy group is, for example, an esterifying group, for example a methyl or an ethyl group which may be removed, for example, by hydrolysis with a base such as sodium hydroxide, or for example a i-butyl group which may be removed, for example, by treatment with an acid, for example an organic acid such as trifluoroacetic acid, or for example a benzyl group which may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.
[00103] Resins may also be used as a protecting group.
[00104] In a particular aspect, the present invention provides a process for the production of a compound of the formula I, or a pharmaceutically acceptable salt or solvate thereof, which comprises:
the reaction of a compou
Figure imgf000035_0001
A
wherein X, Ri, R2, R3, R4, R5, R6, R7, Rs, R21, R22 and R23 each have any one of the meanings defined hereinbefore;
with a compound of formula:
Q-L where Q has any one of the definitions set out hereinbefore and L is a leaving group; and optionally thereafter, and if necessary:
(i) removing any protecting groups present;
(ii) converting the compound formula I into another compound of formula I; and/or
(iii) forming a pharmaceutically acceptable salt or solvate thereof.
[00105] Suitably L is halo (e.g. fluoro, chloro or bromo, particularly chloro or bromo) or another suitable leaving group such as trifluoromethanesulfonate, Meerwein salt or methyl sulfate.
[00106] The reaction is carried out in the presence of a suitable solvent. Any suitable solvent or solvent mixture may be used for this reaction. A person skilled in the art will know how to select suitable solvents or solvent mixtures for use in these reactions. Particular examples of suitable solvents for this reaction include dichloromethane or chloroform.
[00107] Suitably, the reaction is carried out in a sealed vessel.
[00108] Suitably, the reaction is carried out in the presence of a carbonate, for example, potassium carbonate.
[00109] The reaction is carried out an elevated temperature, suitably within the range of 60 to 160 °C or, more suitably 100 to 140 °C, for a suitable time period of, for example, 6 hours to 2 days. A person skilled in the art will be able to select appropriate reaction conditions to use in order to facilitate this reaction.
[00110] The resultant compound of formula I can be isolated and purified using techniques well known in the art.
[00111] As stated above, the compound of formula I may, if necessary, be further reacted to:
(i) remove any protecting groups present;
(ii) convert the compound formula I into another compound of formula I; and/or
(iii) forming a pharmaceutically acceptable salt or solvate thereof.
[00112] An example of (ii) above is when one or more of the groups Ri to R8 may be subject to a further reaction that changes the nature of the substituent group and thereby provides an alternative compound of formula I.
[00113] The process defined herein may further comprise the step of subjecting the compound of formula I to a salt exchange, particularly in situations where the compound of formula I is formed as a mixture of different salt forms. The salt exchange suitably comprises immobilising the compound of formula I on a suitable solid support or resin, and eluting the compounds with an appropriate acid to yield a single salt of the compound of formula I.
[00114] The compounds of formula A can be prepared by processes known in the art.
[00115] The compound of formula A can be prepared by the intra-molecular cyclisation of a compound of formula B
Figure imgf000037_0001
B
wherein X, Ri, R2, R3, R4, R5, R6, R7, Rs, R21, R22 and R23 each have any one of the meanings defined hereinbefore, and L2 is a leaving group (e.g. bromine);
[00116] Suitably, L2 is fluoro, chloro or bromo, particularly chloro or bromo or another suitable leaving group such as trifluoromthanesulfonate.
[00117] In an embodiment, the intra-molecular cyclisation reaction is a palladium catalysed reaction which is suitably carried out using a strong base (such as sodium ierf-butoxide) and a ligand such as PhDavePhos (2-Diphenylphosphino-2'-(N,N-dimethylamino)biphenyl).
[00118] Any suitable solvent may be used for the cyclisation reaction. Examples of suitable solvents include dimethylacetamide (DMA) and dimethylformamide (DMF).
[00119] A person skilled in the art will be able to select appropriate experimental conditions for the cyclisation reaction. Suitably the reaction mixture is heated in some way. In a particular embodiment, the reaction is heated in a microwave for between 5 minutes and 30 minutes.
[00120] The resultant compound of formula A can be isolated and purified using techniques well known in the art.
[00121] In a particular embodiment, the compound of formula B is prepared by reacting a compound of formula C:
Figure imgf000037_0002
wherein X, R4, R5, Re, R7 and Rg are as hereinbefore defined;
a compound of formula D
Figure imgf000038_0001
D
wherein L, R1 ; R2 and R3 are as hereinbefore defined.
[00122] The reaction may be carried out in any suitable solvent. A person skilled in the art will know how to select suitable solvents or solvent mixtures for use in this reaction. Particular examples of suitable solvents include toluene and pyridine.
[00123] The reaction is suitably carried out in the presence of a Lewis acid, such as titanium tetrachloride.
[00124] Suitably, the reaction is carried out in anhydrous conditions and in the presence of an inert atmosphere, such as argon or nitrogen.
[00125] The reaction may be carried out at room temperature or at an elevated temperature, for example at the reflux temperature of the solvent used.
[00126] A person skilled in the art will be able to select appropriate reaction conditions to use to facilitate this reaction.
[00127] The resultant compound of formula B can be isolated and purified using techniques well known in the art.
[00128] Alterntaively, a compound of formula A can be prepared by the intra-molecular cyclisation of a compound of formula E
Figure imgf000039_0001
E
wherein X, Ri, R2, R3, R4, R5, R6, R7, Rs, R21, R22 and R23 each have any one of the meanings defined hereinbefore; Y is halo and Z is a leaving group.
[00129] Suitably Y is bromo .
[00130] Z may be any suitable suitable leaving group. In an embodiment, Z is fluoro.
[00131] In an embodiment, the intra-molecular cyclisation reaction takes place in the presence of an organolithium reagent such as t-butyl-lithium.
[00132] Any suitable solvent may be used for the cyclisation reaction. Examples of suitable solvents include THF.
[00133] A person skilled in the art will be able to select appropriate experimental conditions for the cyclisation reaction. In a particular embodiment, the reaction is treated with i-butyl-lithium at low temperature.
[00134] The resultant compound of formula A can be isolated and purified using techniques well known in the art.
[00135] In a further aspect of the invention, there is provided a compound of formula I obtainable by a process as defined herein.
[00136] In a further aspect of the invention, there is provided a compound of formula I obtained by process as defined herein.
[00137] In a further aspect of the invention, there is provided a compound of formula I directly obtained by process as defined herein.
Biological Activity
[00138] The following biological assays may be used to measure the pharmacological effects of the compounds of the present invention. Surface Plasmon Resonance Assay (SPR)
[00139] Biosensor experiments were conducted in filtered, degassed HEPES buffer (10 mM HEPES, 100 mM KC1, 3 mM EDTA, 0.000 05 v/v of 10% P20 BIACORE surfactant, pH 7.3) at 25 °C. The 5'-biotin labeled DNA sequences (Integrated DNA Technologies) were HPLC
5' 3'
purified and comprise the following sequences: hTel, biotin-d[AG3(T2AG3)3] ; duplex,
^biotindtCGAgTaCiTCTGAgTaCG]3'. The experiments were conducted upon a BIAcore 2000 optical biosensor instrument (BIAcore Inc.). Flow cell 1 was left blank as a reference, while flow cells 2-4 were immobilized with DNA on a streptavidin-derivatized gold chip (SA chip from BIAcore) by manual injection of DNA stock solutions (flow rate of 1 //L/min) until the desired value of DNA response was obtained (350-400 RU). Typically, a series of different ligand concentrations (1 nM to 10 μΜ from 20 mM DMSO stock) were injected onto the chip (flow rate of 50 //L/min, 5- 10 min) until a constant steady-state response was obtained followed by a dissociation period (buffer, 10 min). After every cycle, the chip surface was regenerated (20 s injection of 10 mM glycine solution, pH 2.0) followed by running buffer flow. The data were processed using BIAevaluation (BIAcore Inc.) and Kaleidagraph (Synergy Software) software for nonlinear least-squares optimization of the binding parameters.
Growth Inhibition Assay
[00140] Growth inhibition was measured in the HT-29: human adenocarcinoma cell line using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay:
[00141] Briefly, between 1500 and 2200 cells were seeded into the wells of 96-well microtiter plates and allowed to attach overnight. Test compounds were solubilised at lOmM in dimethyl sulfoxide (DMSO), diluted in tissue culture media and added to cells at final concentrations of 30, 10, 3, 1, 0.3, 0.1 , 0.03 and 0.01 μΜ in triplicate. Following an incubation period of 96 hr MTT was added to a final concentration of 400 g/ml and the plates incubated for a further 4 hr. The media/MTT was then aspirated and 150 μΐ DMSO added. Plates were shaken briefly and the absorbance at 550nm read using a Labtech LT-4000 plate reader. An extra plate was seeded and the MTT assay was carried out on this plate at the time of drug addition to ascertain an initial absorbance value.
[00142] Dose response curves were plotted and the GI50 values (concentration required to inhibit cell growth by 50%) determined for each agent where the absorbance value for control cells minus the initial absorbance was taken as 100% growth. [00143] Although the pharmacological properties of the compounds of the formula I vary with structural change as expected, compounds of the formula I, were found to be active in the above assays.
[00144] In general, the compounds of formula I demonstrate a quadraplexrduplex ratio in the SPR assay of greater than 2.
[00145] By way of example, the quadraplex: duplex ratio in the SPR assay of the compound of Example 2 herein is greater than 15.
[00146] In general, antiproliferative activity possessed by compounds of the formula I, may be demonstrated in the growth inhibition assay by a GI50 value of less than 40 μΜ (preferred compounds have an GI50 of less than 15μΜ, more preferably less than 10μΜ).
[00147] The activity for the following compounds was observed when measured in the growth inhibition and SPR assay described above:
Example HT 29 GI50 (μΜ) SPR (quadruplex Hu SPR (Duplex
Tel (KxlO6 M 1)) (KxlO6 M"1))
1 0.13 1.0 0.1
2 0.21 1.5 <0.1
3 4.39 No binding detected No binding detected
4 0.10
5 0.07
6 24.08
7 0.30
8 0.38 16 0.4
9 7.21 0.4 0.2
10 0.28 12.5 0.1
11 0.35 21.2 0.9
12 2.05
13 0.15 5.2 1.6
14 0.51
15 16.74
16 0.90
17 0.70
18 2.51 68.6 <0.1
19 0.58
20 0.36 35.4 <0.1
21 0.08 56.2 0.6
22 1.14 33.2 0.8
23 0.93 94.4 1.4
24 0.09 11.1 0.2
25 0.50
26 0.58 1.9 0.2
27 0.07 6.2 0.3
28 21.46
29 0.64
30 2.32
31 0.07 14.7 1
32 0.67 Example HT 29 GI50 (μΜ) SPR (quadruplex Hu SPR (Duplex
Tel (KxlO6 M"1)) (KxlO6 M 1))
33 0.13
34 6.07
35 1.23
36 3.11 21.9 0.8
37 0.76 2 0.1
38 0.99
39 0.97
40 0.09 10.7 0.9
41 18.65
42 10.06
43 0.04
44 2.14
45 7.21
46 0.12
47 7.51
48 11.29 17.9 0.1
49 19.92
50 11.25
51 8.01
52 1.39
53 0.31
54 0.24 14.3 <0.1
55 7.16
56 8.70 9.0 0.2
57 2.59 32.3 0.2
58 16.63
59 0.73 5.9 0.5
60 2.21
61 5.89
62 0.138
63 >30
64 5.191
65 0.17
66 1.17
67 0.84
68 5.76
69 0.56
70 1.16
71 8.01
72 4.06
73 2.85
74 2.95
75 0.29
76 2.20
77 >30
78 5.41
79 26.09
80 1.55
81 3.41
82 11.98
83 0.29 Example HT 29 GI50 (μΜ) SPR (quadruplex Hu SPR (Duplex
Tel (KxlO6 M"1)) (KxlO6 M 1))
84 1.30
85 0.54
86 4.29
87 0.20
88 0.18
89 1.35
90 0.14
91 8.37
92 1.68
93 9.80
94 0.14
95 3.43
96 3.17
97 1.24
98 0.66
99 0.83
[00148] The following compounds did not show an activity of 30μΜ or less in the HT 29 assay described above:
chromeno[4,3,2-g/z]phenanthridine 13-oxide;
3,9-dihydroxy-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
3,9-bis(methoxycarbonyl)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium; and
l,ll-difluoro-2-hydroxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium.
[00149] In an embodiment, the compound is not one the above-identified compounds. Pharmaceutical Compositions
[00150] According to a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the invention as defined hereinbefore, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable diluent or carrier.
[00151] The compositions of the invention may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular, intraperitoneal or intramuscular dosing or as a suppository for rectal dosing). [00152] The compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and/or preservative agents.
[00153] An effective amount of a compound of the present invention for use in therapy of infection is an amount sufficient to symptomatically relieve in a warm-blooded animal, particularly a human the symptoms of infection, to slow the progression of infection, or to reduce in patients with symptoms of infection the risk of getting worse.
[00154] The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans will generally contain, for example, from 0.5 mg to 0.5 g of active agent (more suitably from 0.5 to 100 mg, for example from 1 to 30 mg) compounded with an appropriate and convenient amount of excipients which may vary from about 5 to about 98 percent by weight of the total composition.
[00155] The size of the dose for therapeutic or prophylactic purposes of a compound of the invention will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine.
[00156] In using a compound of the invention for therapeutic or prophylactic purposes it will generally be administered so that a daily dose in the range, for example, 0.1 mg/kg to 75 mg/kg body weight is received, given if required in divided doses. In general lower doses will be administered when a parenteral route is employed. Thus, for example, for intravenous or intraperitoneal administration, a dose in the range, for example, 0.1 mg kg to 30 mg kg body weight will generally be used. Similarly, for administration by inhalation, a dose in the range, for example, 0.05 mg/kg to 25 mg kg body weight will be used. Oral administration may also be suitable, particularly in tablet form. Typically, unit dosage forms will contain about 0.5 mg to 0.5 g of a compound of this invention. Therapeutic Uses and Applications
[00157] In one aspect, the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for use as a medicament.
[00158] The compounds of the invention are capable of inhibiting telomerase activity. Thus, in another aspect, the present invention provides a method of inhibiting telomerase activity in a cell, the method comprising administering to said cell compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
[00159] In a further aspect, the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for the inhibition of telomerase in a cell (in vivo or ex vivo) .
[00160] In another aspect, the present invention provides a method of inhibiting telomerase activity in a human or animal subject in need of such inhibition, the method comprising administering to said subject an effective amount of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
[00161] In another aspect, the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof for use in the treatment of disease or condition associated with telomerase activity.
[00162] In another aspect, the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for use in the treatment of disease or condition associated with telomerase activity.
[00163] In yet another aspect, the present invention provides a method of treating a proliferative disorder in a human or animal subject, the method comprising administering to said subject a therapeutically acceptable amount of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
[00164] In yet another aspect, the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for use in the treatment of a proliferative disorder.
[00165] In yet another aspect, the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for use in the treatment of a proliferative disorder.
[00166] The term "proliferative disorder" are used interchangeably herein and pertain to an unwanted or uncontrolled cellular proliferation of excessive or abnormal cells which is undesired, such as, neoplastic or hyperplastic growth, whether in vitro or in vivo. Examples of proliferative conditions include, but are not limited to, pre-malignant and malignant cellular proliferation, including but not limited to, malignant neoplasms and tumours, cancers, leukemias, psoriasis, bone diseases, fibroproliferative disorders (e.g., of connective tissues), and
atherosclerosis. Any type of cell may be treated, including but not limited to, lung, colon, breast, ovarian, prostate, liver, pancreas, brain, and skin. [00167] The anti-proliferative effects of the compounds of the present invention have particular application in the treatment of human cancers by virtue of their telomerase inhibitory properties.
[00168] The anti-cancer effect may arise through one or more mechanisms, including but not limited to, the regulation of cell proliferation, the inhibition of angiogenesis (the formation of new blood vessels), the inhibition of metastasis (the spread of a tumour from its origin), the inhibition of invasion (the spread of tumour cells into neighbouring normal structures), or the promotion of apoptosis (programmed cell death).
[00169] The invention further provides a method of treatment of the human or animal body, the method comprising administering to a subject in need of treatment a therapeutically-effective amount of an active compound, preferably in the form of a pharmaceutical composition.
[00170] Telomerase targeting agents are also known to be useful for inhibiting cancer stem cells (see, for example, US2008/0279961).
[00171] Accordingly, the present invention also provides a method of inhibiting a cancer stem cell, the method comprising administering to said cell an effective amount of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof.
[00172] In another aspect, the present invention provides a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, for use in the inhibition and/or treatment of cancer stem cells.
[00173] In another aspect, the present invention provides the use of a compound of the invention as defined herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for use in the inhibition and/or treatment of cancer stem cells.
Routes of Administration
[00174] The compounds of the invention or pharmaceutical composition comprising the active compound may be administered to a subject by any convenient route of administration, whether systemically/ peripherally or topically (ie., at the site of desired action).
[00175] Routes of administration include, but are not limited to, oral (e.g, by ingestion); buccal; sublingual; transdermal (including, e.g., by a patch, plaster, etc.); transmucosal (including, e.g., by a patch, plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by eyedrops); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., via an aerosol, e.g., through the mouth or nose); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, infraarterlal, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal; by implant of a depot or reservoir, for example, subcutaneously or intramuscularly. Combination Therapies
[00176] The antiproliferative treatment defined hereinbefore may be applied as a sole therapy or may involve, in addition to the compound of the invention, conventional surgery or radiotherapy or chemotherapy. Such chemotherapy may include one or more of the following categories of anti-tumour agents :-
(i) other antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology, such as alkylating agents (for example cis-platin, oxaliplatin, carboplatin,
cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan, temozolamide and nitrosoureas); antimetabolites (for example gemcitabine and antifolates such as
fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea); antitumour antibiotics (for example anthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic agents (for example vinca alkaloids like vincristine, vinblastine, vindesine and vinorelbine and taxoids like taxol and taxotere and polokinase inhibitors);
topoisomerase inhibitors (for example irinotecan, etoposide, teniposide, amsacrine, topotecan and camptothecin) and PARP inhibitors;
(ii) cytostatic agents such as antioestrogens (for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene), antiandrogens (for example bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin), progestogens (for example megestrol acetate), aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5a- reductase such as finasteride;
(iii) anti-invasion agents [for example c-Src kinase family inhibitors like 4-(6-chloro-2,3- methylenedioxyanilino)-7-[2-(4-methylpiperazin-l-yl)ethoxy]-5-tetrahydropyran-4- yloxyquinazoline (AZD0530; International Patent Application WO 01/94341), N-(2-chloro-6- methylphenyl)-2-{ 6-[4-(2-hydroxyethyl)piperazin-l-yl]-2-methylpyrimidin-4-ylamino}thiazole- 5-carboxamide (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661) and bosutinib (SKI-606), and metalloproteinase inhibitors like marimastat, inhibitors of urokinase plasminogen activator receptor function or antibodies to Heparanase] ;
(iv) inhibitors of growth factor function: for example such inhibitors include growth factor antibodies and growth factor receptor antibodies (for example the anti-erbB2 antibody trastuzumab [Herceptin™] , the anti-EGFR antibody panitumumab, the anti-erbBl antibody cetuximab [Erbitux, C225] and any growth factor or growth factor receptor antibodies disclosed by Stern et al. Critical reviews in oncology/haematology, 2005, Vol. 54, ppl l-29); such inhibitors also include tyrosine kinase inhibitors, for example inhibitors of the epidermal growth factor family (for example EGFR family tyrosine kinase inhibitors such as N-(3-chloro-4- fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, ZD1839), N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) and 6- acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib); inhibitors of the hepatocyte growth factor family; inhibitors of the insulin growth factor family; inhibitors of the platelet-derived growth factor family such as imatinib and/or nilotinib (AMN107); inhibitors of serine/threonine kinases (for example Ras/Raf signalling inhibitors such as farnesyl transferase inhibitors, for example sorafenib (BAY 43-9006), tipifarnib (Rl 15777) and lonafarnib (SCH66336)), inhibitors of cell signalling through MEK and/or AKT kinases, c-kit inhibitors, abl kinase inhibitors, PI3 kinase inhibitors, Plt3 kinase inhibitors, CSF-1R kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors; aurora kinase inhibitors (for example AZD1152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 AND AX39459) and cyclin dependent kinase inhibitors such as CDK2 and/or CDK4 inhibitors;
(v) antiangiogenic agents such as those which inhibit the effects of vascular endothelial growth factor, [for example the anti-vascular endothelial cell growth factor antibody bevacizumab (Avastin™) and for example, a VEGF receptor tyrosine kinase inhibitor such as vandetanib (ZD6474), vatalanib (PTK787), sunitinib (SU11248), axitinib (AG-013736), pazopanib (GW 786034) and 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-l- ylpropoxy)quinazoline (AZD2171; Example 240 within WO 00/47212), compounds such as those disclosed in International Patent Applications W097/22596, WO 97/30035, WO 97/32856 and WO 98/13354 and compounds that work by other mechanisms (for example linomide, inhibitors of integrin ανβ3 function and angiostatin)];
(vi) vascular damaging agents such as Combretastatin A4 and compounds disclosed in International Patent Applications WO 99/02166, WO 00/40529, WO 00/41669,
WO 01/92224, WO 02/04434 and WO 02/08213;
(vii) an endothelin receptor antagonist, for example zibotentan (ZD4054) or atrasentan;
(viii) antisense therapies, for example those which are directed to the targets listed above, such as ISIS 2503, an anti-ras antisense;
(ix) gene therapy approaches, including for example approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT (gene-directed enzyme pro-drug therapy) approaches such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multi-drug resistance gene therapy; and
(x) immunotherapy approaches, including for example ex-vivo and in-vivo approaches to increase the immunogenicity of patient tumour cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor, approaches to decrease T-cell anergy, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumour cell lines and approaches using anti-idiotypic antibodies.
[00177] Particular combinations of interest include the compounds of the invention with a topioisomerase inhibitor (such as irinotecan) and/or a PARP inhibitor.
[00178] Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment. Such combination products employ the compounds of this invention within the dosage range described hereinbefore and the other pharmaceutically- active agent within its approved dosage range.
[00179] According to this aspect of the invention there is provided a combination suitable for use in the treatment of a cancer (for example a cancer involving a solid tumour) comprising a compound of the invention as defined hereinbefore, or a pharmaceutically acceptable salt or solvate thereof, and another anti-tumour agent.
[00180] According to this aspect of the invention there is provided a combination suitable for use in the treatment of a cancer (for example a cancer involving a solid tumour) comprising a compound of the invention as defined hereinbefore, or a pharmaceutically acceptable salt or solvate thereof, and any one of the anti-tumour agents listed under (i) - (ix) above.
[00181] In a further aspect of the invention there is provided a compound of the invention or a pharmaceutically acceptable salt or solvate thereof, in combination with an anti-tumour agent selected from one listed under (i) - (ix) herein above.
[00182] Herein, where the term "combination" is used it is to be understood that this refers to simultaneous, separate or sequential administration. In one aspect of the invention "combination" refers to simultaneous administration. In another aspect of the invention "combination" refers to separate administration. In a further aspect of the invention "combination" refers to sequential administration. Where the administration is sequential or separate, the delay in administering the second component should not be such as to lose the beneficial effect of the combination.
[00183] According to a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the invention, or a pharmaceutically acceptable salt or solvate thereof in combination with an anti-tumour agent selected from one listed under (i) - (ix) herein above, in association with a pharmaceutically acceptable diluent or carrier.
Examples
[00184] The invention will now be illustrated in the following Examples in which, generally:
(i) Unless stated otherwise, operations were carried out at ambient temperature, i.e. in the range 17 to 25 °C and optionally under an atmosphere of an inert gas such as nitrogen or argon.
(ii) In general, the course of the reactions described herein were followed by liquid chromatography mass spectrometry (LCMS). The reaction times that are given are not necessarily the minimum attainable.
(iii) When necessary, organic solutions were dried over anhydrous magnesium sulfate, work-up procedures were carried out using traditional layer separating techniques, evaporations were carried out by rotary evaporation in vacuo.
(iv) Yields, where present, are not necessarily the maximum attainable, and when necessary, reactions were repeated if a larger amount of the reaction product was required.
(v) In general, the structures of the end-products were confirmed by nuclear magnetic resonance (NMR) and/or mass spectral techniques.
LCMS data were obtained using an Agilent 1200 series LC/MS with an Agilent 6110 quadropole MS, with Electrospray ionisation. The following three methods were used:
(a) Polar method:
Mobile phase A - 0.1% Acetic acid in water. Mobile Phase B- 0.1% acetic acid in acetonitrile. Flow rate of l.OOml/min. Gradient from 2% B over 3 minutes to 20%B for 0.2 minutes, up to 95% B for 1.8 minutes and then back down to 2%B. Total run time of 6 minutes. Column:
Phenomenex Gemini-NX 3μπι C18, 30 x 2.00mm. Chromatograms based on UV detection at 254nm.
Mass Spectra were achieved using the MS in positive mode.
(b) Alkaline method:
Mobile phase A- 0.1% ammonia solution in water. Mobile phase B- 0.1% ammonia solution in acetonitrile. Flow rate of 0.500ml/min. Gradient from 5% B over 3 minutes to 95%B, remaining at 95%B until 4.1 minutes, when it returns down to 5%B until the 7 minute run is over.
Column: Waters XBridge3^m C18 20 X 2.1mm IS column. Chromatograms based on UV detection at 254nm. Mass Spectra were achieved using the MS in positive mode
Generally, only ions relating to the parent structure are reported.
(c) Trial method: Mobile phase A - 0.1% Acetic acid in water. Mobile Phase B- 0.1% in acetonitrile. Flow rate of l.OOml/min. Gradient from 5% B rising up to 95% B over 3 minutes, remaining at 95%B for 1 minute and then back down to 5% B over 6 seconds. The total run time is 5 minutes.
Column: Phenomenex Gemini-NX 3μπι CI 8, 30 x 2.00mm. Chromatograms based on UV detection at 254nm.
Mass Spectra were achieved using the MS in positive mode.
Proton NMR chemical shift values were measured on the delta scale at 300 or 400 MHz using a Bruker AV400. The following abbreviations have been used: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad. Coupling constants are reported in Hz
(vi) Unless stated otherwise, compounds containing an asymmetric carbon atom were not resolved.
(vii) Intermediates were not necessarily fully purified but their structures and purity were assessed by TLC, analytical HPLC, infra-red (IR) and/or NMR analysis;
(viii) Unless otherwise stated, column chromatography (by the flash procedure) and medium pressure liquid chromatography (MPLC) were performed on Merck Kieselgel silica (Art. 9385).
(ix) The following analytical HPLC methods were used:
- in general, reversed-phase silica was used with a flow rate of about 1 ml per minute and detection was by Electrospray Mass Spectrometry and by UV absorbance at a wavelength of 254 nm.
(x) Compounds were purified by flash silica chromatography using Merck Kieselgel silica (Art. 9385).
Note: The synthesis of substituted chromeno[4,3,2-g/z]phenanthridines and thiochromeno[4,3,2- gA]phenanthridines was adapted from the procedures reported in J. Org. Chem. 2003, 68, 4091- 4092
Preparation of Basic Building Blocks
Preparation of polymer supported trialkyl ammonium salts resins:
A suspension of polymer supported trialkyl ammonium resin, hydroxide form (amberlyst A26, HO-) (4g) in 0.1M methanesulfonic acid (40mL) was stirred for lOmin. A small excess of 1M methanesulfonic acid was added until the pH of the solution was acidic. The mixture was stirred for 5min., then filtered. The resin was washed with water until the pH of filtrate was neutral. The resin was then washed successively with methanol, 90% DCM:MeOH and dried under suction to give polymer supported trialkyl ammonium mesylate form. The same process was used to prepare polymer supported trialkyl ammonium formate form using formic acid instead of methanesulfonic acid.
Preparation of silica bound trimethyl ammonium salts resins:
Silica bound trimethylammonium chloride resin (1 g) was stirred in degassed water and was filtered. This process was repeated three times and the resin was added to 1L of a 0.1M aqueous solution of 2:1 methanesulfonic acid: sodium methanesulfonate. The mixture was stirred for 10 min. and decantated. This was repeated two more times and the silica was filtered and transferred to a small column. The silica bound resin was washed successively with two column volume of 0.1M aqueous solution of 2:1 methanesulfonic acid: sodium methanesulfonate, two column volume of 0.01M aqueous solution of 2:1 methanesulfonic acid: sodium methanesulfonate, and two column volume of water to give silica bound trimethyl ammonium mesylate.
The same process was used to prepare polymer supported trialkyl ammonium formate, tosylate and acetate using the corresponding 2:1 acid:base buffer.
Preparation of N-(prop-2-ynyl)acetamide
Figure imgf000052_0001
Py, DMAP, DCM
Acetic anhydride (3.24 mL, 34.3 mmol, 1.1 equ) was added dropwise to a solution of propargyl amine (2 mL, 31.2 mmol), pyridine (2.76 mL, 34.3 mmol, 1.1 equ) and N,N dimethylaminopyridine (few crystals, catalytic amount) in dichloromethane (50 mL) at 0°C and the mixture was stirred overnight. The mixture was diluted with dichloromethane and transferred into a separating funnel. Water was added and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate. The product was absorbed on silica and purified by flash chromatography (Hexane:EtOAc 30%-20 ) to give the title compound as an off white crystalline solid (1.45 g, 49% yield).
<¾ (CDC13): 5.82 (1H, s (br)), 4.04-4.06 (2H, dd, J=5.3, 2.6), 2.23-2.24 (1H, t, J=2.6), 2.01 (3H, s).
Preparation of 3,3-dimethyldioxirane
Figure imgf000052_0002
The synthesis of 3,3-dimethyldioxirane was carried out according to the following procedure - Organic Syntheses, 1998, Vol. 74, p. 91 (1997); Coll. Vol. 9, p.288
A 2-L, three-necked, round-bottomed flask containing a mixture of water (80 mL), acetone (50 mL, 0.68 mol), and sodium bicarbonate (96 g), was equipped with a magnetic stirring bar and a pressure equalizing addition funnel containing water (60 mL) and acetone (60 mL, 0.82 mol). An air condenser (50 cm length) loosely packed with glass wool was attached to the reaction vessel. The outlet of the air condenser was connected to a Dewar condenser filled with dry ice-acetone connected to a receiving flask (250 mL) cooled in a dry ice-acetone bath. The receiving flask was also connected in series to a second dry ice-acetone cold trap, a trap containing potassium iodide solution, and a vacuum was placed on the apparatus. Oxone was added in portions (10-15 g) over approximately 0.5h while the acetone-water mixture was simultaneously added dropwise and the reaction mixture was stirred vigorously throughout the addition of reagents. A yellow solution of 3,3-dimethyldioxirane in acetone collected in the receiving flask. Vigorous stirring was continued for an additional 15 min while a vacuum was applied to the cold trap. The yellow dioxirane solution was dried over sodium sulfate, filtered and stored in the freezer (-25 °C) over activated molecular sieves.
The overall yield was 65mL.
Example 1 - Preparation of 13-methyIthiochromeno[4 ..2-gfe1phenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation o -bromo-N-(9H-thioxanthen-9-ylidene)aniline
Figure imgf000053_0001
A solution of titanium tetrachloride (776 ί, 7.08 mmol, 0.75 equ) in toluene (8 mL) was added dropwise to a suspension of thioxanthone (2.0 g, 9.42 mmol) and 2-bromoaniline (4.8 mL, 42.5 mmol, 4.5 equ) in toluene (24 mL) at 0°C. The mixture was stirred at 0°C for 30 min. The mixture was allowed to warm to room temperature and was refluxed overnight. The resulting black suspension was cooled to room temperature and filtered over Celite. The filtrate was concentrated to dryness in vacuo and the resulting solid was dissolved in DCM. The product was absorbed on silica and purified by flash chromatography (gradient elution hexane:EtOAc 90%- 80%) to give the title compound as a yellow/orange solid (1.47 g, 43% yield). <¾ (CDCI3): 8.51 (1H, s (br)), 7.63-7.66 (1H, dd, J=8.1, 1.3), 7.54-7.56 (2H, d, J=8.6), 7.42 (3H, m (br)), 7.15-7.19 (1H, ddd, J=7.8, 7.5, 1.3), 7.15 (2H, m (br)), 6.92-6.97 (1H, ddd, J=7.9, 7.4, 1.6), 6.62-6.64 (1H, dd, J=7.9, 1.6).
m/z (ES+): 366.0 (MH+, 79Br), 368.0 (MH+, 81Br).
Step 2-Preparation
Figure imgf000054_0001
A mixture of 2-bromo-N-(9H-thioxanthen-9-ylidene)aniline (200 mg, 0.546 mmol), sodium tert- butoxide (105 mg, 1.09 mmol, 2 equ), 2-Diphenylphosphino-2'-(N,N-dimethylamino)biphenyl (21 mg, 0.0546 mmol, 0.1 equ), and PdCl2(PPh3)2 (~l mg, catalytic amount) in DMA (4 mL) was heated at 200°C under microwave irradiation (300W, 300psi) for 5min. (5min. hold time with constant cooling). Four consecutive runs were performed and the combined reactions mixtures were poured into ice. The resulting precipitate was filtered and dried under suction. The product was dissolved in DCM, absorbed on silica and purified by flash chromatography (gradient elution Hexane:EtOAc 95%-90%).The fractions containing the pure product were collected, while the impure fractions were absorbed on silica and re -purified by flash chromatography (Hexane: EtOAc 95%). The combined pure fractions were concentrated in vacuo to give the title compound as a yellow solid (250 mg, 40% yield).
<¾ (CDCI3): 9.16 (1H, s (br)), 8.42-8.44 (1H, m), 8.32-8.35 (1H, m), 8.19 (1H, s(br)), 7.69-7.75 (2H, m), 7.55-7.61 (2H, m), 7.44-7.46 (2H, m), 7.35-7.39 (1H, m).
m/z (ES+): 286.1(MH+).
Step 3 - Preparation of 13-methylthiochromeno[4,3,2-gft]phenanthridin-13-ium
trifluoromethanesulfonate
Figure imgf000054_0002
A suspension of thiochromeno[4,3,2-g/z]phenanthridine (250 mg, 0.876 mmol), methyl triflate (300 \L, 2.73 mmol, 3.1 equ) and potassium carbonate (-300 mg, -2.17 mmol, -2.5 equ) in CHCI3 (10 mL) was heated at 100°C in a sealed tube overnight. The reaction was quenched with methanol and was transferred into a flask. A saturated solution of sodium bicarbonate was added slowly with stirring and the product was extracted with DCM. The combined organic extracts were dried over magnesium sulfate, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as an orange/red solid (350 mg, 89% yield).
<¼ (DMSO-ifc): 8.97-8.99 (1H, dd, J=8.2, 1.3), 8.92-8.94 (1H, dd, J=8.4, 1.0), 8.42-8.46 (2H, m), 8.35-8.37 (1H, dd, J=7.8, 1.0), 8.25-8.29 (1H, dd, J=8.0), 8.15-8.17 (1H, dd, J=8.2, 7.0), 8.07- 8.12 (1H, ddd, J=8.6, 7.2, 1.4), 7.95-7.99 (1H, m), 7.91-7.95 (1H, ddd, J=8.3, 7.2, 1.2), 7.74- 7.78 (1H, ddd, J=8.4, 7.2, 1.2), 4.55 (3H, s).
m/z (ES+): 300.1 (M+).
Example 2 - Preparation of 13-methyIchromeno[4,3,2-jg¾1phenanthridin-13-ium
methanesulfonate
Step 1 - Preparation of -bromo-N-(9H-xanthen-9-ylidene)aniline
Figure imgf000055_0001
A solution of titanium(IV) chloride (422 μΐ^, 3.82 mmol, 0.75 equ) in toluene (5 mL) was slowly added to a solution of 9H-xanthen-9-one (1.00 g, 5.1 mmol) and 2-bromoaniline (3.945 g, 22.9 mmol, 4.5 equ) in toluene (15 mL) under nitrogen at 0°C. The resulting mixture was stirred for 30 min at ambient temperature then a condenser was fitted and the reaction was heated to reflux. After heating for 16 hours, the reaction was cooled to room temperature when diethyl ether (50 mL) was added, and the reaction mixture was passed through a pad of Celite with diethyl ether (50 mL x 3) washings. The solvent was removed under reduced pressure and the resulting solid was recrystallised using toluene and hexanes which was collected by filtration and washed with hexanes to give the title compound as a white solid (1.059 g, 60% yield).
<¾ (DMSO- e): 8.20-8.22 (1H, dd, J=8.0, 1.7), 7.87-7.92 (1H, ddd, J=8.0, 1.7, 1.4), 7.68-7.72
(2H, dd, J=8.0, 1.2), 7.52-7.54 (2H, d, J=8.4), 7.36-7.40 (2H, dt, J=7.7, 1.3), 7.02-7.06 (2H, ddd,
J=8.8, 1.6, 1.0), 6.91-6.94 (2H, dd, J=7.9, 1.5).
m/z (ES+): 350 +(MH+, 79Br), 352 (MH+, 81Br).
Step 2 - Preparation of chromeno[4,3,2-g i]phenanthridine
Figure imgf000056_0001
A microwave vial was charged with 2-bromo-N-(9H-xanthen-9-ylidene)aniline (250 mg, 1.43 mmol), sodium ie/t-butoxide (274 mg, 2.86 mmol, 2.0 equ), a catalytic amount of tris(dibenzylideneacetone)dipalladium, followed by a catalytic amount of diphenylphosphino-2- (N,N-dimethylamino)biphenyl. The RM was diluted with anhydrous dimethylacetamide (5 mL) and then heated under microwave radiation at 180°C (300W, 200psi, 5 mins). This reaction was repeated in duplicate. The combined crude reaction mixtures were filtered and the residual solid was washed with EtOAc. The combined organics were washed with saturated brine (1 x 25 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. The crude material was then subjected to flash silica chromatography (gradient elution EtOAc:Hexanes 100%-99%-98%- 97%-96%-95%) to give the title compound as a yellow solid (368 mg, 55% yield).
<¾ (DMSO-de): 8.65-8.67 (1H, dd, J=8.2, 1.4), 8.58-8.60 (1H, dd, J=7.9, 1.4), 8.43-8.44 (1H, d, J=7.8), 8.03-8.05 (1H, dd, J=8.2, 0.9), 7.95-7.99 (1H, t, J=8.1), 7.74-7.78 (1H, ddd, J=8.2, 7.0, 1.3), 7.62-7.69, (2H, m), 7.52-7.54 (1H, dd, J=8.0, 0.8), 7.46-7.48 (1H, dd, J=8.3, 0.7), 7.40-7.46 (1H, ddd, J=8.0, 7.2, 1.2).
m/z (ES+): 270 (M+H+).
Step 3 - Preparation of 13-methylchromeno[4,3,2-g i]phenanthridin-13-ium
methanesulfonate MsO
Figure imgf000056_0002
A solution of chromeno[4,3,2-g/i]phenanthridine (92 mg, 0.34 mmol, Example 6) in anhydrous chloroform (10 ml) in a sealed tube was stirred with potassium carbonate (48 mg, 0.34, 1 equ) and methyl trifluoromethanesulfonate (774 μL·, 7.06 mmol, 20 equ) and then heated to 100°C. Every 24 h a further amount of potassium carbonate (0.048 g, 0.34 mmol, 1 equ) and methyl trifluoromethanesulfonate (774 μί, 7.06 mmol, 20 equ) was added and after 120 h the reaction was quenched with MeOH and saturated sodium bicarbonate solution (10 mL) was added slowly with stirring and the crude product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo. After absorbing on silica, the crude product was purified by flash silica chromatography (gradient elution DCM:MeOH 99%-98%-97%-95%) to yield a yellow solid.
The resulting solid (70 mg) was then stirred in DCM:MeOH (10:1, 30 mL) when amberlyst mesylate resin (700 mg) was added. After 16 h, the reaction mixture was filtered, concentrated in vacuo and the residue sonicated in Et20/hexanes (10 mL) to yield a yellow/orange suspension which was collected by filtration and air dried to give the title compound as a yellow/orange solid (56 mg, 43% yield)
<¾ (DMSO-J6): 8.93-8.95 (1H, dd, J=8.3, 1.4), 8.76-8.78 (1H, d, J=8.0), 8.58-8.60 (1H, dd, J=8.5, 1.4), 8.40-8.44 (1H, t, J=8.5), 8.39-8.42 (1H, d, J=8.8), 8.05-8.12, (2H, m), 8.01-8.03 (1H, d, J=8.3), 7.91-7.97 (2H, m), 7.69-7.73 (1H, ddd, J=8.4, 7.3, 1.2), 4.64 (3H, s), 2.28 (3H, s). m/z (ES+): 284 (M +).
Example 3 - Preparation of 13-methylthiochromeno[4,3.i2-gfelphenanthridin-13-ium-10,10- dioxide trifluoromethanesulfonate
Figure imgf000057_0001
To a solution of 13-methylthiochromeno[4,3,2- /i]phenanthridin-13-ium
trifluoromethanesulfonate (25 mg, 0.056 mmol Example 1) in chloroform (5 mL) was added 3- chloroperoxybenzoic acid (70%) (30 mg, 0.12 mmol, 2.2 equ) and the reaction was stirred at RT. After 16 h, a further amount of 3-chloroperoxybenzoic acid (70%) (200 mg, 0.17 mmol, 15 equ) and after stirring for a further 24 h, the reaction was diluted with saturated NaHCC>3 solution (lOmL) and extracted with DCM. The combined organics were dried over magnesium sulfate, filtered and concentrated in vacuo. After absorbing on silica, the crude product was purified by flash silica chromatography (gradient elution DCM:MeOH,100%-98%-95%) to give the title compound as a light brown solid (6 mg, 22% yield).
m/z (ES+): 318 (M+). Example 4 - Preparation of 2-(furan-2-yl)-13-methylchromeno[4,3,2-g¾lphenanthridin-13- ium methanesulfonate
Step 1 - Preparation of 2',3-difluoro-4'-methoxybiphenyl-2-carbonitrile
Figure imgf000058_0001
A suspension of 2-Bromo-6-fluorobenzonitrile (540 mg, 2.7 mmol), 2-fluoro-4- methoxyphenylboronic acid (550 mg, 3.2 mmol, 1.2 equ), potassium fluoride (510 mg, 0.8 mmol, 3.3 equ) and bis[tris(t-butyl)phosphine]palladium(0) (13 mg, 2 mol%) in anhydrous tetrahydrofuran (15 mL) was heated for 2 hrs under an atmosphere of nitrogen at 60°C. After cooling, saturated brine (10 mL) was added, and the aqueous layer extracted with dichloromethane (2 x 10 mL). The extracts were dried over sodium sulphate, evaporated to dryness in vacuo and the residue subjected to flash chromatography (chloroform) to give the title compound as a white solid (620 mg, 94% yield).
4 (CDCL): 7.64 (1H, ddd, J= 2.0, 6.0, 8.4), 7.38 - 7.28 (2H, m), 7.20 (1H, t, J= 8.0), 6.85 (1H, dd, J= 2.0, 8.0), 6.80 (1H, dd, J= 3.0, 7.8), 3.88 (3H, s).
m/z (ES+): 246.1 (MH+).
Step 2 - Preparation of 3-(2-bromophenoxy)-2'-fluoro-4'-methoxybiphenyl-2-carbonitrile
Figure imgf000058_0002
Potassium carbonate (113 mg, 0.8 mmol, 2 equ) was added to a solution of 2',3-difluoro-4'- methoxybiphenyl-2-carbonitrile (100 mg, 0.4 mmol) and 2-bromophenol (105 mg, 0.6 mmol, 1.5 equ) in anhydrous N,N-dimethylformamide (2 mL) at 25°C. The reaction mixture was heated at 100°C for 24 hrs and was allowed to cool. A saturated ammonium chloride solution (10 mL) was added, and the aqueous layer extracted with diethyl ether (6 x 10 mL). The extracts were washed with saturated brine (5 mL), dried over sodium sulfate and evaporated to dryness. Recrystallisation from ethyl acetate / heptane gave the title compound as a tan coloured solid (140 mg, 86% yield).
<¾ (CDCL): 7.72 (1H, dd, J= 1.6, 8.0), 7.50 - 7.38 (3H, m), 7.20 - 7.16 (3H, m), 6.85 (1H, dd, J= 2.0, 8.0), 6.80 (1H, dd, J= 2.8, 8.4), 6.66 (1H, d, J= 8.4), 3.89 (3H, s).
m/z (ES+): 398.0 (MH+, 79Br), 400.0 (MH+, 81Br). - Preparation of 2-methoxychromeno[4,3,2-g¾]phenanthridine
Figure imgf000059_0001
A solution of 3-(2-bromophenoxy)-2'-fluoro-4'-methoxybiphenyl-2-carbonitrile (690 mg, 1.7 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise to a solution of tert-butyl lithium (2.27 mL (1.6M pentane), 3.6 mmol, 2.1 equ) in anhydrous tetrahydrofuran (15 mL) at - 78°C. After 5 mins, the temperature was allowed to warm to 25°C and the reaction stirred for another 30 mins. Saturated ammonium chloride (25 mL) was added and the aqueous layer extracted with dichloromethane (3 x 50 mL). The extracts were dried over sodium sulfate and evaporated to dryness. Recrystallisation from ethyl acetate / heptane gave the title compound as a yellow solid (250 mg, 48% yield).
& (CDC ): 8.70 (1H, d, J= 8.0), 8.33 (1H, d, J= 9.2), 8.07 (1H, d, J= 8.0), 7.80 (1H, d, J= 8.0), 7.54 (2H, m), 7.37 - 7.29 (3H, m), 7.20 (1H, dd, J= 2.8, 8.8), 4.03 (3H, s).
m/z (ES+): 300.0 (MH+).
Step 4 - Preparation of chromeno[4,3,2-g/i]phenanthridin-2-yl trifluoromethanesulfonate
Figure imgf000059_0002
2-methoxychromeno[4,3,2-g/i]phenanthridine (130 mg, 0.43 mmol) and pyridine hydrochloride (250 mg, 2.1 mmol, 5 equ) were heated under MW irradiation at 215°C for 5min. (300W, lOOpsi.). After cooling, water (5 mL) was added and the resulting precipitate filtered from solution and allowed to dry. The crude phenol (110 mg, 0.38 mmol) was dissolved in a mixture of chloroform (5 mL) and tetrahydrofuran (5 mL). To this solution were added N,N- diisopropylethylamine (100 mg, 0.76 mmol, 2 equ) and N-phenyl bis-trifluoromethane sulfonimide (207 mg, 0.58 mmol, 1.5 equ) and the reaction mixture heated at 50°C for 24 hrs. The reaction mixture was then evaporated to dryness, and purified by flash chromatography (chloroform) to give the title compound as pale yellow solid (100 mg, 62% yield).
<¼ (CDCI3): 8.71 (1H, dd, J= 1.6, 8.0), 8.48 (1H, d, J= 8.8), 8.14 (1H, d, J= 8.0), 8.00 (1H, d, J= 2.8), 7.86 (1H, d, J= 8.0), 7.60 (1H, t, J= 8.4), 7.48-7.28 (4H, m). m/z (ES+): 418.0 (MH ).
Step 5 - Preparation of 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2- g¾]phenanthridin-13-
Figure imgf000060_0001
A mixture of chromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate (260 mg, 0.624 mmol), methyl triflate (4.35 g, 26.5 mmol, 42 equ), potassium carbonate (400 mg, 2.89 mmol, 4.6 equ) in chloroform (40 mL) was heated in a sealed tube at 150 °C for 2 h. The reaction mixture was dissolved in 95% DCM:MeOH and purified by flash column chromatography (gradient elution DCM:MeOH 95%-88%) to give the title compound as a yellow solid (256 mg, 0.440 mmol, 71 % yield) .
<¾ (DMSO- d6): 8.83-8.81 (1H, d, J= 9.1), 8.56-8.54 (1H, dd, J= 7.8, 1.6), 8.45-8.43 (1H, d, J= 7.8), 8.02-7.98 (2H, m), 7.73-7.68 (2H, m), 7.59-7.57 (1H, dd, J= 8.1, 0.5), 7.49-7.47 (1H, dd, J= 8.3, 0.8), 7.45-7.41 (1H, ddd, J= 8.0, 7.2, 1.1).
m/z (ES+): 432.1 (M+)
Step 6 - Preparation of 2-(furan-2-yl)-13-methylchromeno[4,3,2- ¾]phenanthridin-13-ium methanesulfonate
Figure imgf000060_0002
A mixture 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.240 mmol), tetrakis(triphenylphosphine)palladium(0) (27 mg, 0.0240 mmol, 0.1 equ), potassium carbonate (67 mg, 0.480 mmol, 2 equ) and 2-furanyl boronic acid (54 mg, 0.480 mmol, 2 equ) in 1: 1 DME:H20 (6 mL) was heated at 80°C under microwave irradiation (300 W, 300 psi) for 10 min. The reaction mixture was concentrated in vacuo, dissolved in 95% DCM:MeOH and purified by flash column chromatography (gradient elution DCM:MeOH 98%-90%). The combined pure fractions were concentrated in vacuo, and dissolved in DCM:MeOH 90% (10 mL). This was stirred with silica bound trimethylammonium- mesylate (400 mg) for 2 h and filtered through Celite. The filtrate was evaporated to dryness to give the title compound as a yellow solid (39 mg, 0.0875 mmol, 36% yield).
4 (DMSO- d6): 9.00-8.95 (1H, d, J= 8.7), 8.77-8.75 (1H, d, J= 8.0), 8.59-8.57 (1H, d, J= 8.0), 8.49 (1H, s), 8.43-8.40 (1H, dd, J= 8.2, 8.2), 8.28-8.26 (1H, d, J = 8.5), 8.13-8.09 (1H, dd, J= 7.5, 7.5), 8.01-7.99 (1H, 2H, m), 7.94-7.92 (1H, d, J = 8.1), 7.76-7.72 (1H, dd, J=7.6, 7.6), 7.54-7.53 (1H, d, J= 3.4), 6.81-6.80 (1H, dd, J= 3.4, 1.7), 4.71 (3H, s(br)), 2.29 (3H, s(br)).
m/z (ES+): 350.1 (M+)
Example 5 - Preparation of 2-(furan-3-yl)-13-methylchromeno[4,3,2-g¾lphenanthridin-13- ium methanesulfonate
Figure imgf000061_0001
A mixture 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.240 mmol; Example 4), tetrakis(triphenylphosphine)palladium(0) (27 mg, 0.0240 mmol, 0.1 equ), potassium carbonate (67 mg, 0.480 mmol, 2 equ) and 3-furanyl boronic acid (54 mg, 0.480 mmol, 2 equ) in 1 :1 DME:H20 (6 mL) was heated at 80°C under microwave irradiation (300 W, 300 psi) for 10 min. The reaction mixture was concentrated in vacuo, dissolved in 95% DCM:MeOH and purified by flash column chromatography (gradient elution DCM:MeOH 98%-90%). The combined pure fractions were concentrated in vacuo, dissolved in DCM:MeOH 90% (10 mL) and stirred with silica bound trimethylammonium-mesylate (750 mg) for 2 h. The mixture was filtered through Celite and the filtrate was evaporated to dryness to give the title compound as a yellow solid (86 mg, 0.193 mmol, 80% yield).
<¼ (DMSO- d6): 8.93-8.91 (1H, d, J= 8.6), 8.77-8.75 (1H, d, J= 8.0), 8.67 (1H, s), 8.58-8.56 (1H, d, J= 7.8), 8.45 (1H, s), 8.42-8.38 (1H, dd, J= 8.2,8.2), 8.23-8.21 (1H, d, J= 8.5), 8.11-8.08 (1H, dd, J= 7.4, 7.4), 7.99-7.97 (1H, d, J= 8.2)m 7.92-7.91 (2H, m), 7.75-7.71 (1H, dd, J = 8.0, 8.0), 7.40-7.40 (1H, d, J= 1.0), 4.70 (3H, s(br)), 2.29 (3H, s(br)).
m/z (ES+): 350.1 (M+) Example 6 - Preparation of 2-hvdroxy-13-methylchromeno[4,3^2-g¾lphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000062_0001
Sodium feri-butoxide (500 mg, 5.20 mmol, 10.1 equ) was added to a stirring solution of 13- methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/z]phenanthridin-13-ium
trifluoromethanesulfonate (300 mg, 0.516 mmol; Example 4) in water (20 mL) and methanol (20 mL) and heated at reflux for 2 h. The reaction mixture was allowed to cool to room temperature and 6 M HCl added dropwise until pH 1. The mixture was concentrated in vacuo, dissolved in 94% DCM:MeOH (8 mL), filtered through Celite and purified by flash column chromatography (gradient elution DCMrMeOH 94%-90%) to give the title compound as a brown solid (214 mg, 0.476 mmol, 92% yield).
<¾ (DMSO- d6): <¾ (DMSO- d6): 11.13 (1H, s(br)), 8.77-8.75 (1H, d, J= 9.1), 8.56-8.55 (1H, d, J= 7.8), 8.56-8.53 (1H, dd, J= 8.4,1.2), 8.34-8.29 (1H, dd, J= 8.2, 8.2), 8.08-8.07 (1H, ddd, J= 8.6, 7.2, 1.3), 7.88-7.86 (1H, dd, J= 8.5,1.1), 7.84-7.82 (1H, dd, J= 8.5, 0.5), 7.70-7.65 (1H, ddd, J= 8.3, 7.2, 1.1), 7.60-7.60 (1H, d, J= 2.2), 7.41-7.39 (1H, dd, J= 8.8, 2.1), 4.55 (3H, s(br)).
m/z (ES+): 300.1 (M+)
Note: 2-hydroxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium was also obtained as a byproduct of palladium catalysed coupling reactions of 13-methyl-2- (trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium
trifluoromethanesulfonate. The pure product was obtained as the hemicarbonate salt after purification of the mixture.
Example 7 - Preparation of 2-(2-hvdroxyethoxy)-13-methylchromenor4,,3..2- gfelphenanthridin-13-ium methanesulfonate
Figure imgf000062_0002
2-Bromoethanol (454 mg, 3.63 mmol, 10 equ) was added dropwise to a stirring suspension of 2- hydroxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium hemicarbonate (120 mg, 0.363 mmol) and potassium carbonate (502 mg, 3.63 mmol, 10 equ) in anhydrous N,N- dimethylformamide (5 mL). The reaction mixture was heated at 100°C for 1 h and allowed to cool to room temperature. 90% DCM:MeOH (15 mL) was added and the mixture was filtered through Celite. Acetic acid was added to the filtrate until acidic pH was attained and the mixture was concentrated in vacuo. The reaction mixture was dissolved in DCM:MeOH 90% (8 mL) and purified by column chromatography (gradient elution DCM:MeOH 90%-80%). The combined pure fractions were concentrated in vacuo, dissolved in 90% DCM:MeOH (10 mL) and was stirred with silica bound trimethylammonium-mesylate (1.5 g) for 2 h. The mixture was filtered through Celite and the filtrate was evaporated to dryness in vacuo to give the title compound as a yellow solid (145 mg, 0.330 mmol, 91% yield).
4 (DMSO- d6): 8.85-8.83 (1H, d, J= 9.2), 8.64-8.62 (1H, d, J= 7.8), 8.55-8.53 (1H, dd, 8.4, 1.2), 8.37-8.32 (1H, dd, 8.2, 8.2), 8.09-8.05 (1H, ddd, J= 8.4, 7.2, 1.3), 7.90-7.88 (1H, dd, J= 8.4, 1.1), 7.89-7.87 (1H, dd. J= 8.2, 0.5), 7.75-7.75 (1H, d, J= 2.3), 7.72-7.68 (1H, ddd, J= 8.4, 7.2, 1.2), 7.60-7.57 (1H, dd, J= 9.0, 2.3), 5.06-5.04 (1H, t, J= 5.4), 4.61 (3H, s(br)), 4.38-4.35 (2H, t, J=4.8), 3.88-3.85 (2H, dt, J=5.2, 4.5), 2.29 (3H, s(br)).
m/z (ES+): 344.1 (M+) Examples 8 and 9 - Preparation of 2-acetamido-13-methylchromeno[4,3,2- g½1phenanthridin-13-ium trifluoromethanesulfonate (Example 8); andl3-methyl-2-(N- methylacetamido)chromeno[4,3.l2-g¾lphenanthridin-13-ium 2/3 trifluoromethanesulfonate
1/3 acetate (Example 9)
Step 1 - Preparation of 2-bromo-5-nitro-N-(9H-xanthen-9-ylidene)aniline
Figure imgf000063_0001
2-Bromo-5-nitroaniline (4.43 g, 20.4 mmol, 2 equ) was added portionwise to a stirring solution of xanthone (2.00 g, 10.2 mmol), 1.0 M titanium chloride in toluene (10.2 mL, 10.2 mmol, 1 equ) and N,N-diisopropylethylamine (13.2 g, 102 mmol, 10 equ) in toluene (300 mL). The mixture was refluxed under nitrogen for 2 h and 1.0 M titanium chloride in toluene (10.2 mL, 10.2 mmol, 1 equ) was added. The reaction mixture was refluxed under nitrogen for 16 h and was allowed to cool to room temperature. Methanol (10 mL) followed by 95% DCM:MeOH (200 mL) were added and the mixture was filtered through Celite. The filtrate was concentrated in vacuo and the residue was dissolved in dichloromethane, dry loaded onto silica and purified by column chromatography (gradient elution Hex:EtOAc 98%-95%) to give the title compound as a pale brown solid (2.70 g, 6.83 mmol, 67% yield).
<¼ (DMSO- d6): 7.97-8.23 (1H, m (br)), 8.06-8.06 (1H, d, J= 9.0), 7.83-7.86 (1H, dd, J- 8.8, 2.7), 7.79 (1H, d, J= 2.5), 7.70-7.74 (2H, m), 7.57-7.59 (2H, d, J=8.4), 7.26 (3H, m (br)).
m z (ES+): 395.0 (MH+, 79Br), 397.0 (MH+, 81Br) Remaining Steps - Preparation of 2-acetamido-13-methylchromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate (Example 8) and 13-methyl-2-(N- methylacetamido)chromeno[4,3,2-g/j]phenanthridin-13-ium 2/3 trifluoromethanesulfonate 1/3 acetate (Example 9)
Figure imgf000064_0001
A mixture of 2-bromo-5-nitro-N-(9H-xanthen-9-ylidene)aniline (100 mg, 0.253 mmol), tris(dibenzylideneacetone)dipalladium(0) (10 mg, 0.0127, 0.05 equ), 1,8- diazabicyclo[5.4.0]undec-7-ene (190 mg, 1.27 mmol, 5 equ) and 2-diphenylphosphino-2'-(N,N- dimethylamino)biphenyl (10 mg, 0.0253 mmol, 0.1 equ) in dimethylacetamide (9 mL) was heated at 200°C under microwave irradiation (300 W, 200 Psi) for 5 min. This was repeated nine times and the combined reaction mixtures were concentrated in vacuo. The mixture was dissolved in 98% DCM:MeOH (100 mL) and filtered through celite. The mixture was concentrated in vacuo to give 2-nitrochromeno[4,3,2-gft]phenanthridine (m/z (ES+): 315.1 (MH+)), which was used without further purification in the next step.
A mixture of 2-nitrochromeno[4,3,2-g i]phenanthridine (318 mg, 1.01 mmol) and tin chloride dihydrate (1.14 g, 5.05 mmol, 5 equ) in ethanol (35 mL) was refluxed under nitrogen for 16 h. The reaction mixture was allowed to cool to room temperature and concentrated in vacuo. The dry residue was sonicated in 1 M sodium hydroxide (aq) (50 mL) and the resulting precipitate was filtered. The filtrate was extracted with dichloromethane (2 x 50 mL) and the extracts and solid precipitate combined, adding more dichloromethane (20 mL) to ensure complete dissolution. The mixture was dry loaded onto silica and purified by flash column chromatography (gradient elution DCM:MeOH 100%-97%). The combined pure fractions were concentrated in vacuo to give chromeno[4,3,2-g z]phenanthridin-2-amine (m/z (ES+): 285.1 (MH+) as a pale yellow solid (96 mg, 0.378 mmol, 37% yield). The product was used without further characterization in the next step.
Phthaloyl dichlonde (88 mg, 0.422 mmol, 1.5 equ) was added to a stirring solution of chromeno[4,3,2-g/z]phenanthridin-2-amine (80 mg, 0.282 mmol), pyridine (112 mg, 1.408 mmol, 5 equ) and N, N dimethylaminopyridine (~1 mg, catalytic amount) in dichloromethane (40 mL). The reaction mixture was allowed to stir under nitrogen at room temperature for 10 min and then concentrated in vacuo. The solid residue was triturated with 0.5 M HC1 (aq) (20 mL) and the resulting precipitate was collected by filtration. The filtrate was extracted with dichloromethane (2 x 20 mL) and the organic extracts were concentrated in vacuo and combined with the precipitate yielding 130 mg of 2-(chromeno[4,3,2-g i]phenanthridin-2-yl)isoindoline-l,3-dione (m/z (ES+): 415.1 (MH+)) as a yellow solid (130 mg, 100% yield). The product was used without further purification and characterization in the next step.
A mixture of 2-(chromeno[4,3,2-g i]phenanthridin-2-yl)isoindoline-l,3-dione (130 mg, 0.317 mmol), methyl triflate (520 mg, 3.17 mmol, 10 equ) and potassium carbonate (428 mg, 3.17 mmol, 10 equ) in chloroform (3 mL) was heated in a sealed tube at 110 °C for 16 h. The reaction mixture was allowed to cool to room temperature and methanol (2 mL) was added. DCM:MeOH 90% (10 mL) was added and the mixture filtered and concentrated in vacuo to give a mixture of 2-(l,3-dioxoisoindolin-2-yl)-13-methylchromeno[4,3,2-gA]phenanthridin-13-ium
trifluoromethanesulfonate (m/z (ES+): 429.1 (M+)), 2-(2-(methoxycarbonyl)-N- methylbenzamido)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium
trifluoromethanesulfonate (m/z (ES+): 475.5 (M+)) and 2-(dimethylamino)-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate (m/z (ES+): 327.2 (M+)) as identified by LCMS analysis. This mixture of product was used without further purification in the next step.
A I M solution of hydrazine monohydrate in ethanol (0.18 mL, 0.476 mmol, 1.5 equ) was added dropwise to a stirring solution of the mixture obtained above (183 mg, 0.317 mmol) in ethanol (5 mL) and heated at reflux for 1 h. The reaction mixture was allowed to cool to room temperature and concentrated in vacuo. The residue was triturated with a saturated sodium bicarbonate solution (50 mL) and extracted with dichloromethane (2 x 50 mL). The combined organic layers were absorbed on silica and purified by flash column chromatography (gradient elution DCM:MeOH 99%-90%) to give two main fractions.
The fractions containing pure 2-amino-13-methylchromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (m/z (ES+): 299.1 (M+)) were concentrated in vacuo to give 111 mg of a yellow solid. The product was used without further characterization in the next step.
The fractions containing an unseparable mixture of 13-methyl-2-(methylamino)chromeno[4,3,2- g/j]phenanthridin-13-ium trifluoromethanesulfonate and 2-(dimethylamino)-13- methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate were concentrated in vacuo to give 21 mg of a yellow solid. This mixture of product was used without further purification in the next step.
Acetyl chloride (22 mg, 0.280 mmol, 6.3 equ) was added to a stirring solution of 2-amino-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate (20 mg, 0.0446 mmol) and pyridine (24 mg, 0.281 mmol, 6.3 equ) in dichloromethane (2 mL) for 1 h at room temperature. The reaction mixture was evaporated to dryness in vacuo, dissolved in DCM/MeOH 90% (3 mL) and purified by flash column chromatography (gradient elution 90%- 85%). The combined pure fractions were concentrated in vacuo to give 2-acetamido-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate as a yellow solid (5 mg, 0.0102 mmol, 23% yield).
<¼ (CDC13): 10.2 (1H, s), 8.90-8.87 (1H, dd, J= 9.0, 1.7). 8.48-8.46 (1H, d, J= 7.4), 8.45 (1H, s), 8.39-8.39 (1H, dd, J=8.4, 1.2), 8.37-8.34 (1H, d, J= 8.4), 8.27-8.23 (1H, dd, J= 8.2, 8.2), 8.01- 7.97 (1H, ddd, J= 8.6, 7.3, 1.4), 7.78-7.75 (1H, dd, J= 8.5, 1.0), 7.74-7.71 (1H, dd, J= 8.1, 0.8), 7.67-7.65 (1H, ddd, J- 8.4, 7.2, 1.2), 4.77 (3H, s(br)), 2.29 (3H, s(br)).
m/z (ES+): 341.1 (M+)
Acetyl chloride (8 mg, 0.0983 mmol) was added to the mixture of 13-methyl-2- (methylamino)chromeno[4,3,2- z]phenanthridin-13-ium trifluoromethanesulfonate and 2- (dimethylamino)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate obtained above (21 mg) and pyridine (11 mg, 0.131 mmol) in dichloromethane (2 mL) and stirred under nitrogen at room temperature for 1 h. The reaction mixture was concentrated in vacuo and dissolved in DCM:MeOH 90% (5 mL) and purified by flash column chromatography (gradient elution DCM:MeOH 90%-86%) giving two separate product fractions. The first combined pure fractions were concentrated in vacuo to give 13-mefhyl-2-(N- methylacetamido)chromeno[4,3,2-g/z]phenanthridin-13-ium 1/3 trifluoromethanesulfonate 2/3 acetate as a yellow solid (10 mg, 0.0198 mmol).
4 (CDC13): 8.64-8.61 (1H, dd, J- 8.7, 0.9), 8.56-8.54 (1H, d, J= 8.7), 8.38-8.36 (1H, d, J- 7.9), 8.26-8.22 (1H, dd, J= 8.2, 8.2), 8.17 (1H, s(br)), 7.98-7.94 (1H, ddd, J= 8.5, 7.2, 1.4), 7.78-7.76 (1H, d, J= 7.8), 7.73-7.67 (3H, m), 4.77 (3H, s(br)), 3.51 (3H, s(br)), 2.18 (3H, s(), 1.74 (3H, s(br)).
m z (ES+): 355.1 (M+)
Note: Only traces of (dimethylamino)-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (m/z (ES+): 327.2 (M+)) were isolated after purification.
Alternatively, 2-acetamido- 13-methylchromeno[4,3,2-gA]phenanthridin- 13-ium methanesulfonate could be obtained by palladium catalysed cross-coupling of acetamide with 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2- z]phenanthridin-13-ium
trifluoromethanesulfonate:
Figure imgf000067_0001
A mixture of 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g i]phenanthridin-13- ium trifluoromethanesulfonate (100 mg, 0.240 mmol, Example 4), tris(dibenzylideneacetone)dipalladium(0) (6 mg, 0.012 mmol, 0.05 equ), 2,2'- bis(diphenylphosphino)-l,l'-binaphthyl (4 mg, 0.024 mmol, 0.1 equ), acetamide (71 mg, 1.20 mmol, 5 equ) and cesium carbonate (235 mg, 0.720 mmol, 3 equ) in 1,4-dioxane (8 mL) was heated at 80 °C under microwave irradiation (300 W, 300 psi) for 2 min. A second portion of caesium carbonate (14 mg, 0.240 mmol, 3 equ) was added and the mixture was heated at 80 °C under microwave irradiation (300 W, 300 psi) for 5 min. The reaction mixture was filtered through Celite, evaporated to dryness in vacuo, dissolved in DCM:MeOH 95% (4 mL) and purified by flash column chromatography (gradient elution DCM:MeOH 95%-90%). The combined pure fractions were concentrated in vacuo and the resulting orange solid was then stirred in DCM:MeOH (90%, 20 ml) when polymer supported trialkylammonium mesylate resin (0.500 g) was added. After lh, the reaction mixture was filtered, concentrated in vacuo and the residue triturated in Et20/hexanes (20 mL) to give the title compound the title compound as yellow/orange solid (21 mg, 0.0481 mmol, 28% yield).
4 (CDCI3): 11.25 (1H, s), 9.00-8.97 (1H, dd, J= 9.1, 1.8), 8.83-8.82 (1H, d, J- 1.7), 8.46-8.44 (1H, d, J= 9.2), 8.41-8.39 (1H, dd, J=8.5, 1.3), 8.35-8.33 (1H, dd, J= 8.3,0.5), 8.25-8.21 (1H, t, J= 8.2,8.2), 7.99-7.94 (1H, ddd, J= 8.6, 7.2, 1.4), 7.76-7.73 (1H, dd, J= 8.5, 1.1), 7.71-7.69 (1H, dd, J= 8.1, 0.8), 7.65-7.61 (1H, ddd, J= 8.4, 7.2, 1.2), 4.86 (3H, s(br)), 2.94 (3H, s(br)), 2.35 (3H, s(br)).
m/z (ES+): 341.1 (M+), Example 10 - Preparation of 2-amino-13-methylthiochromeno[4,3,2-jg¾1phenanthridin-13- ium trifluoromethanesulfonate
Figure imgf000068_0001
To a stirred solution of 2-bromo-5-nitroaniline (1.00 g, 4.61 mmol, 3 equ) and 9H-thioxanthen-9- one (326 mg, 1.54 mmol) in anhydrous toluene (10 mL) was added a solution of titanium chloride (83 μL·, 0.77 mmol, 0.5 equ) in toluene (3 mL). The mixture was refluxed under nitrogen for 72 h when 2-bromo-5-nitroaniline (500 mg, 2.30 mmol, 1.5 equ) was added followed by a further solution of solution of titanium chloride (42 μΐ^, 0.39 mmol, 0.25 equ) in toluene (1.5 mL) was added. The reaction mixture was refluxed under nitrogen for 6 h when a further solution of solution of titanium chloride (42 μΐ^, 0.39 mmol, 0.25 equ) in toluene (1.5 mL) was added. After stirring for 16 h, the reaction was allowed to cool to room temperature, when toluene was added and the mixture was filtered through Celite. The filtrate was concentrated in vacuo and the residue was dissolved in dichloromethane, dry loaded onto silica and purified by column chromatography (gradient elution He :Et20 90%) to give 2-bromo-5- nitro-N-(9H-thioxanthen-9-ylidene)aniline m/z (ES+): 411.0 (MH+, 79Br), 413.0 (MH+, 81Br) as a yellow solid (163 mg, 26% yield) which was used without further characterisation in the next step.
A mixture of 2-bromo-5-nitro-N-(9H-thioxanthen-9-ylidene)aniline (100 mg, 0.244 mmol), palladium acetate (8.6 mg, 0.012, 0.1 equ), sodium carbonate (28.4 mg, 0.268 mmol, 1.1 equ) and 2-diphenylphosphino-2'-(N,N-dimethylamino)biphenyl (9.4 mg, 0.0244 mmol, 0.1 equ) in dimethylacetamide (8 mL) was heated at 180°C under microwave irradiation (300 W, 200 Psi) for 5 min. This was repeated nine times and the combined reaction mixtures were concentrated in vacuo, absorbed on silica and purified by flash chromatography (gradient elution Hexane:DCM 60%-50%-40%-0% then DCM:MeOH 99%-98%) to give 2- nitrothiochromeno[4,3,2-g z]phenanthridine m/z (ES+): 331.1 (MH+) as an orange solid (613 mg) which was used without further purification in the next step.
A mixture of 2-nitrothiochromeno[4,3,2-g/z]phenanthridine (613 mg, 1.85 mmol) and tin chloride dihydrate (1.67 g, 7.42 mmol, 4 equ) in ethanol (60 mL) was refluxed under nitrogen for 16 h. The reaction mixture was allowed to cool to room temperature and concentrated in vacuo. The dry residue was sonicated in 1M sodium hydroxide and then extracted with dichloromethane (2 x 50 mL). The combined organic extracts were dried over magnesium sulfate, filtered then concentrated in vacuo. The mixture was dry loaded onto silica and purified by flash column chromatography (gradient elution DCM:MeOH 100%-99%-98%) to give thiochromeno [4,3,2- g i]phenanthridin-2-amine (m z (ES+): 301.1 (MH+) as a green/brown solid (96 mg, 35% yield over 2 steps). The product was used without further characterization in the next step.
A suspension of phthalic anhydride (30 mg, 0.200mmol, 1.2 equ) in chloroform (1 mL) was added to a suspension of thiochromeno[4,3,2-g/?]phenanthridin-2-amine (50 mg, 0.166 mmol) in chloroform (5 mL) and the mixture was refluxed overnig ht, then heated at 85°C for 4 h. The mixture was concentrated to dryness and the resulting brown solid was triturated with sat. NaHCC>3 and the precipitate was filtered, then dried under suction. The product was triturated with acetone and the precipitate was filtered. The product, which was obtained as a -1:1 mixture with uncyclized phthalimide (2-(thiochromeno[4,3,2-g/z]phenanthridin-2- ylcarbamoyl)benzoic acid) as shown by LCMS analysis, was used without purification in next step.
A suspension of 2-(thiochromeno[4,3,2-g¾]phenanthridin-2-yl)isoindoline-l,3-dione (77 mg, 0.179 mmol), methyl triflate (196 \\L, 1.78 mmol, 10 equ), and potassium carbonate (246 mg, 1.78 mmol, 10 equ) in chloroform (5 mL) was heated at 110°C in a sealed tube overnight. The reaction was quenched with methanol and the insoluble, inorganic materials were removed by filtration. The filtrate was absorbed on silica and the product was purified by flash chromatography (gradient elution DCM:MeOH 96%-94%-92%) to give 30 mg of 2-(l,3- dioxoisoindolin-2-yl)-13-methylthiochromeno[4,3,2-gA]phenanthridin-13-ium
trifluoromethanesulfonate as a -2: 1 mixture with uncyclized phthalimide methyl ester (2-(2- (methoxycarbonyl)benzamido)-13-methylthiochromeno[4,3,2- /z]phenanthridin-13-ium trifluoromethanesulfonate). The product was used without further purification in the next step. A 1M solution of hydrazine monohydrate (76 μί, 0.0757 mmol, 1.5 equ) (1 mL) in ethanol was added at room temperature to a solution of 2-(l,3-dioxoisoindolin-2-yl)-13- methylthiochromeno[4,3,2- z]phenanthridin-13-ium trifluoromethanesulfonate (30 mg, 0.0504 mmol) in ethanol (2 mL) and the reaction was refluxed for lh. The mixture was concentrated to dryness in vacuo and the resulting dark solid was suspended in sat. NaHCCh. The product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate. The product was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 96%-94%-92%) to give 20 mg of 2-amino-13-methylthiochromeno[4,3,2- g j]phenanthridin-13-ium trifluoromethanesulfonateas a dark purple solid.
<¼ (DMSO-i¾): 8.61-8.64 (1H, d, J=9.2), 8.56-8.58 (1H, dd, J=8.2, 1.2), 8.38-8.40 (1H, dd, J=8.3, 0.9), 8.02-8.06 (1H, dd, J1=J2=7.9), 8.03-8.05 (1H, dd, J=8.1, 1.2), 7.99-8.01 (1H, dd, J=7.6, 1.2), 7.81-7.85 (1H, ddd, J=8.3, 7.3, 1.2), 7.65-7.69 (1H, ddd, J=8.4, 7.2, 1.3), 7.21-7.24 (2H, m), 6.70 (2H, s), 4.37 (3H, s).
m/z (ES+): 315.1 (M+)
Note: The NMR spectrum showed that the product was contaminated with some uncharacterized aromatic impurity. The product was used without further purification in the next step.
Acetyl chloride (4.6 \L, 0.0646 mmol, 1.5 equ) was added to a suspension of 2-amino-13- methylthiochromeno[4,3,2- z]phenanthridin-13-ium trifluoromethanesulfonate (20 mg, 0.04431 mmol) and N,N-Diisopropylethylamine (9 \L , 0.0517 mmol, 1.2 equ) in dichloromethane (1 mL) at 0°C and the mixture was stirred at 0°C for 45 min. 9 \iL of N,N-Diisopropylethylamine followed by 65 of a 1M solution of acetyl chloride in dichloromethane were added and the mixture was stirred at 0°C for 1 h. Water was added and the mixture was transferred into a separating funnel. The product was extracted with dichloromethane and the combined organic extracts were washed with sat. NaHCC>3, dried over magnesium sulfate and concentrated in vacuo. The product was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-97.55-96%-95%) to give 13 mg of a dark pink/orange solid. The product was triturated with diethyl ether, filtered and dried under suction to give the title compound as a dark pink/orange solid (8 mg, 8% yield over 4 steps). <¾ (DMSO-J6): 10.8 (1H, s), 8.90-8.92 (1H, d, J=9.1), 8.79-8.80 (1H, d, J=1.8), 8.77-8.79 (1H, dd, J=8.1, 1.2), 8.47-8.50 (1H, dd, J=8.3, 0.9), 8.25-8.27 (1H, dd, J=7.8, 1.1), 8.19-8.23 (1H, dd, J1=J2=7.8), 8.12-8.14 (1H, dd, J=8.1, 1.1), 7.98-8.00 (1H, dd, J=9.0, 1.8), 7.88-7.92 (1H, ddd, J=8.2, 7.2, 1.2), 7.70-7.74 (1H, ddd, J=8.3, 7.3, 1.3), 4.47 (3H, s), 2.22 (3H, s).
Figure imgf000071_0001
m/z (ES+): 357.1 (M+)
Example 11 - Preparation of 13-methyl-2-(lH-pyrazol-4-yl)thiochromenor4,3,2- gfelphenanthridin-13-ium chloride hydrochloride
Step 1 - Preparation of 2
Figure imgf000071_0002
An aqueous solution (2 mL) of sodium nitrite (28 mg, 0.406 mmol, 1.5 equ) was added dropwise at 0°C to a suspension of thiochromeno[4,3,2-g z]phenanthridin-2-amine (80 mg, 0.266 mmol) in 2N HC1 (4 mL). The resulting thick suspension was diluted with water and the mixture was stirred at 0°C for 20min. An aqueous solution (2 mL) of potassium iodide (76 mg, 0.452 mmol, 1.7 equ) was added and the mixture was stirred at 80°C for 30min. The mixture was basified at 0°C to pH 13 with 5N NaOH and a saturated solution of potassium sulfite was added. The product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate then concentrated in vacuo. The product was dissolved in dichloromethane and the solution was loaded on a short column packed with silica. The product was eluted with DCM to give the title compound as a yellow/pale brown solid (75 mg, 69% yield).
<¾ (CDC13): 9.04-9.06 (1H, m), 8.50-8.51 (1H, d, J=1.7), 8.25-8.27 (1H, dd, J=8.4, 1.0),8.10- 8.12 (1H, d, J=8.6), 7.80-7.83 (1H, dd, J=8.6, 1.8), 7.67-7.71 (1H, dd, J=8.1, 7.8), 7.55-7.57 (1H, dd, J=7.6, 1.0), 7.41-7.44 (2H, m), 7.34-7.36 (1H, m).
Step 2 - Preparation of 2-iodo-13-methylthiochromeno[4,3,2-gA]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000072_0001
A suspension of 2-iodothiochromeno[4,3,2-g¾]phenanthridine (75 mg, 0.182 mmol), methyl triflate (200 ί, 1.82 mmol, 10 equ) and potassium carbonate (250 mg, 1.82 mmol, 10 equ) in chloroform (5 mL) was heated at 110°C in a sealed tube for 3 days. The reaction was quenched with methanol and the resulting suspension was filtered. The filtrate was absorbed on silica and the product was purified by flash chromatography (gradient elution DCM:MeOH 98%-96%- 94%-92%-90 ) to give the title compound as a red solid (70 mg, 67% yield).
4 (DMSO-i¾): 8.88-8.90 (1H, dd, J=8.2, 0.8), 8.72 (1H, d, J=1.5), 8.69-8.71 (1H, d, J=8.8), 8.40-8.42 (1H, dd, J=8.4, 0.8), 8.37-8.39 (1H, dd, J=7.8, 0.8), 8.24-8.29 (2H, m), 8.15-8.18 (1H, dd, J=8.1, 1.0), 7.91-7.95 (1H, ddd, J=8.2, 7.2, 1.2), 7.74-7.78 (1H, ddd, J=8.4, 7.2, 1.2), 4.49 (3H, s).
m/z (ES+): 426.0 (M+)
Step 3 - Preparation of 13-methyl-2-(lH-pyrazol-4-yl)thiochromeno[4,3,2-^ i]phenanthridin-13- ium chloride hydrochloride
Figure imgf000072_0002
A mixture of 2-iodo-13-methylthiochromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate (30 mg, 0.0521 mmol), lH-Pyrazole-4-boronic acid pinacol ester (152 mg, 0.781 mmol, 15 equ), tetrakis(triphenylphosphine)palladium(0) (6 mg, 0.00521 mmol, 0.1 equ) and potassium carbonate (14.4 mg, 0.104 mmol, 2 equ) in 1:1 dioxane:H20 (1 mL) was heated at 80°C for lOmin. under MW irradiation (300W, 300psi, hold time lOmin. with cooling system on). The reaction mixture was concentrated to dryness in vacuo and the resulting solid was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-96%-94%-92%-90%) to give 80 mg of a dark red solid. The compound was triturated with 3N HCl and the resulting precipitate was filtered, washed successively with water and diethyl ether and dried under suction to give the title compound as a red solid (13 mg, 56% yield).
4 (DMSO-i¾): 8.93-8.95 (1H, d, J=8.7), 8.88-8.90 (1H, dd, J=8.2, 1.0), 8.49-8.53 (3H, m), 8.41-8.44 (1H, dd, J=8.4, 1.0), 8.28-8.30 (1H, dd, J=7.8, 0.9), 8.24-8.26 (1H, dd, J=8.7, 1.6), 8.20-8.24 (1H, dd, J1=J2=7.8), 8.13-8.16 (1H, dd, J=8.1, 1.1), 7.89-7.93 (1H, ddd, J=8.2, 7.1, 1.2), 7.73-7.78 (1H, ddd, J=8.3, 7.1, 1.2), 4.59 (3H, s).
m/z (ES+): 366.1 (M+)
Note: the 19F NMR spectrum confirmed that the product was obtained as the chloride salt. Example 12 - Preparation of 2-(3-acetamidoprop-l-vnyl)-13-methylthiochromenor4,3.2- gfelphenanthridin- -ium iodide
Figure imgf000073_0001
A mixture of 2-iodo-13-methylthiochromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate (25 mg, 0.0434 mmol; Example 11), N-acetyl propargylamine (21 mg, 0.087 mmol, 5 equ), tetrakis(triphenylphosphine)palladium(0) (2.5 mg, 0.0008 mmol, 0.05 equ) and copper iodide (catalytic amount taken out with the end of a pipette) in 10:1 DMF:piperidine (1 mL) was heated at 80°C for 2min. under MW irradiation (300W, 300psi, hold time 2min. with cooling system on). A reaction mixture from a separate run carried out on 10 mg scale was combined and the mixture was concentrated to dryness in vacuo. The product was triturated with water and the resulting precipitate was filtered, washed with a small amount of sat. NaHC(¾, then water and dried under suction. The product was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-96%-95%-94%- 92%-90%) to give the title compound as a red solid (14 mg, 44% yield).
<¾ (DMSO-ii6): 8.93-8.95 (1H, d, J=8.5), 8.88-8.90 (1H, d, J=7.9), 8.49-8.52 (1H, t (br), J=5.5), 8.36-8.41 (3H, m), 8.25-8.29 (1H, dd, J1=J2=8.0), 8.16-8.18 (1H, dd, J=8.2, 1.0), 7.92-7.95 (2H, m), 7.75-7.78 (1H, m), 4.51 (3H, s), 4.24-4.26 (2H, d, J=5.5), 1.90 (3H, s).
m/z (ES+): 395.1 (M+)
Notel: the 19F NMR spectrum confirmed that the product was obtained as the iodide salt. Note2: To remove any residual silica, the product was dissolved in 95% DCM:MeOH and the resulting solution was filtered over Celite. The filtrate was concentrated in vacuo and the product was triturated with diethyl ether, filtered and dried under suction. Example 13 - Preparation of 13-methyl-2-(thiophen-3-yl)thiochromeno[4,3,2- g½lphenanthridin- -ium trifluoromethanesulfonate
Figure imgf000074_0001
A mixture of 2-iodo-13-methylthiochromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (30 mg, 0.0521 mmol; Example 11), thiophen-3-ylboronic acid (8.0 mg, 0.0626 mmol, 1.2 equ), tetrakis(triphenylphosphine)palladium(0) (6 mg, 0.0052 mmol, 0.1 equ) and potassium carbonate (14.4 mg, 0.104 mmol, 2 equ) in 1:1 dioxane:H20 (1 mL) was heated at 80°C for 10 min. under MW irradiation (300W, 300psi, hold time lOmin. with cooling system on). The reaction mixture was concentrated to dryness in vacuo and the resulting solid was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-96%-95) to give the title compound as a red solid (13 mg, 46% yield).
<¾ (DMSO-c¾: 8.98-9.00 (1H, d, J=8.8), 8.92-8.94 (1H, d, J=8.1), 8.58-8.59 (1H, d, J=1.5), 8.41-8.44 (2H, m), 8.32-8.36 (2H, m), 8.23-8.27 (1H, dd, J1=J2=8.0), 8.15-8.17 (1H, dd, J=8.2, 1.3), 7.97-7.99 (1H, dd, J=5.1, 1.2), 7.90-7.94 (1H, m), 7.83-7.85 (1H, dd, J=5.1, 2.9), 7.75-7.79 (1H, m), 4.62 (3H, s).
Figure imgf000074_0002
m/z (ES+): 382.0 (M+)
Example 14 - Preparation of 6,10-dimethoxy-13-methylchromenor4,3^2-g¾lphenanthridin- 13-ium
Step 1 - Preparation of 3, 6-dihydroxy-9H-xanthen-9-one (synthesized by modification of a procedure reported in Grover, P. K.; Shah, G. D.; Shah, R. C. /. Chem. Soc. 1955, 3982-3985)
Figure imgf000074_0003
A suspension of bis(2,4-dihydroxyphenyl)methanone (6 g, 24.4 mmol) in water was heated at 190°C for 48 hours. The reaction mixture was cooled to room temperature and the resulting precipitate was filtered, washed with acetonitrile and diethyl ether to give the title compound as pale yellow solid (5.56 g, 100% yield).
<¼ (DMSO-d6): 10.81 (2H, s,), 7.97-7.99 (2H, d, J=8.8), 6.85-6.88 (2H, dd, J=8.7, 2.2), 6.82 (2H, d, J=2.2)
m/z (ES+): 229 (MH+), 251 (M+Na+)
Step 2 - Prepa
Figure imgf000075_0001
To a suspension of 3, 6-dihydroxy-9H-xanthen-9-one (5.5 g, 24.1 mmol) in N,N- dimethylformamide was added sodium hydride (60% in mineral oil, 1.01 g, 25.3 mmol, 1.05 equ) portion wise and the mixture was stirred at room temperature for 10 minutes before the addition of methyl iodide (4.70 mL, 50.6 mmol, 2.1 equ). The mixture was stirred at ambient temperature for 18 hours and the resulting suspension was filtered. The solid was washed with water and hexane giving the title compound as a solid (5.6 g, 91% yield).
<¾ (DMSO-d6): 8.07-8.09 (2H, d, J=8.8), 7.10 (2H, d, J=2.3), 7.03-7.06 (2H, dd, J=8.8, 2.4), 3.93 (6H, s).
m/z (ES+): 257 (MH+), 279 (M+Na+)
Step 3 - line
Figure imgf000075_0002
A 1M solution of titanium tetrachloride in toluene (8.78 mL, 8.78 mmol, 1.5 equ) was added dropwise to a suspension of 3,6-dimethoxy-9H-xanthen-9-one (1.5 g, 5.85 mmol), N,N diisopropylethylamine (5.1 mL, 29.2 mmol, 5 equ) and 2-bromoaniline (1.32 mL, 11.7 mmol, 2 equ) in toluene (45 mL) and the resulting orange suspension was refluxed for 2 h. The mixture was filtered through Celite and the filtrate was concentrated in vacuo. The product was dissolved in dichloromethane, absorbed on silica and purified by flash chromatography (Hexane:EtOAc 90:10) to give the title compound as a pale yellow solid (1.28 g, 53% yield).
<¾ (CDC13): 8.43 (1H, m (br)), 7.62-7.64 (1H, dd, J=8.0, 1.3), 7.27-7.32 (1H, m), 7.11-7.15 (1H, m), 6.80-6.97 (5H, m), 6.45-6.55 (1H, m), 3.88 (6H, s). - Preparation of 6,10-dimethoxy-13-methylchromeno[4,3,2-^ft]phenanthridin-13-
Figure imgf000076_0001
A mixture of 2-bromo-N-(3,6-dimethoxy-9H-xanthen-9-ylidene)aniline (450 mg, 1.1 mmol), sodium t-butoxide (211 mg, 2.2 mmol, 2 equ), 2-Diphenylphosphino-2'-(N,N- dimethylamino)biphenyl (42 mg, 0.011 mmol, 0.1 equ), and bis(triphenylphosphine)palladium(II) dichloride (catalytic amount taken with the end of a pipette) in anhydrous dimethylacetamide (5 mL) was heated at 180°C under microwave irradiation for 5min. (300W, 300psi, hold time 5min. with cooling system on). 3 consecutive runs were carried out and the combined reaction mixtures were concentrated to a small volume in vacuo. The residue was poured into ice and the precipitate was filtered, washed with water and dried under suction. The product was dissolved in DCM:MeOH and purified by flash chromatography (Hexane:EtOAc 90% then DCM:MeOH 100%-99%-98%) to give 6,10- dimethoxychromeno[4,3,2-g/z]phenanthridine (650 mg), which was contaminated with unseparable impurities. The product was used without further characterization and purification in the next steps.
A solution of 6,10-dimethoxychromeno[4,3,2-g¾]phenanthridine (150 mg, 0.455 mmol), methyl triflate (500 μΐ^, 4.55 mmol, 10 equ) and potassium carbonate (629 mg, 4.55 mmol, 10 equ) in chloroform (5 mL) was heated at 100°C overnight. The reaction was quenched with methanol and filtered. The filtrate was concentrated in vacuo and the residue was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-96%-95%) to give the title compound as a yellow/green solid (87 mg, 39% yield).
<¼ (DMSO-i¾): 8.87-8.90 (1H, dd, J=8.3, 1.2), 8.45-8.47 (1H, d, J=9.4), 8.23-8.26 (1H, d, J=8.3), 8.19-8.20 (1H, d, J=2.2), 7.97-8.02 (1H, ddd, J=8.5, 7.2, 1.2), 7.81-7.86 (1H, ddd, J=8.0, 7.3, 0.7), 7.52-7.53 (1H, d, J=2.2), 7.37-7.38 (1H, d, J=2.6), 7.26-7.29 (1H, dd, J=9.4, 2.6), 4.49 (3H, s), 4.15 (3H, s), 4.05 (3H, s).
m/z (ES+): 344.1 (M+) Example 15 - Preparation of 6,10-bis(2-hvdroxyethoxy)-13-methylchromeno[4,3.2- gfelphenanthridin-13-ium bromide
Step 1 - Preparation of 6,10-dihydroxy-13-methylchromeno[4,3,2-^]phenanthridin-13- ium bromide
Figure imgf000077_0001
A 1M solution of boron tnbromide in dichloromethane (1 mL) was added to a suspension of 6,10-dimethoxy-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium trifluoromethanesulfonate (77 mg, 0.156 mmol; Example 14) dichloromethane (1 mL) and the mixture was stirred overnight. The reaction was quenched with methanol and the mixture was stirred for ~30min. The mixture was concentrated in vacuo and the product was suspended in DCM:MeOH. The precipitate was filtered, washed with diethyl ether and dried under suction to give the title compound as a bright yellow/green solid (45 mg, 73% yield).
<¾ (DMSO-d6): 11.9 (1H, s (br)), 11.7 (1H, s (br)), 8.61-8.63 (1H, dd, J=8.3, 1.2), 8.36-8.38 (1H, d, J=9.2), 8.17-8.19 (1H, d, J=8.4), 7.93-7.97 (1H, ddd, J=8.5, 7.1, 1.2), 7.90-7.91 (1H, d, J=2.0), 7.76-7.80 (1H, dd, J1=J2=7.3), 7.23-7.24 (1H, d, J=2.0), 7.08-7.11 (1H, dd, J=9.1, 2.5), 7.06- 7.07 (1H, d, J=2.4), 4.42 (3H, s).
m/z (ES+): 316.1 (M+)
Notel: the 19F NMR spectrum confirmed that the product was obtained as the bromide salt. Step 2 - Preparation of 6,10-bis(2-hydroxyethoxy)-13-methylchromeno[4,3,2- gft]phenanthridin-13-ium bromide
Figure imgf000077_0002
A suspension of 6,10-dihydroxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium bromide (20 mg, 0.0505 mmol; ) and potassium carbonate (70 mg, 0.505 mmol, 10 equ) in N,N- dimethylformamide (1 mL) was stirred for 10 min. before the addition of 2-bromoethanol (36\L, 0.505 mmol, 10 equ). The mixture was stirred at 100°C for 2 h and a further lOequ of 2- bromoethanol were added. The mixture was stirred at 100°C for 16 h and an additional lOequ of 2-bromoethanol were added. The mixture was stirred at 100°C for 2 h and was allowed to cool to room temperature. The mixture was diluted with N,N-dimethylformamide and filtered through a syringe filter. The filtrate was concentrated in vacuo and the product was dissolved in DCM:MeOH. The product was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 95%-90 -85%) to give the title compound as a yellow/orange solid (13 mg, 54% yield).
<¼ (DMSO-i¾: 8.87-8.89 (1H, dd, J=8.3, 1.0), 8.44-8.47 (1H, d, J=9.4), 8.22-8.25 (1H, d, J=8.4), 8.19-8.20 (1H, d, J=2.1), 7.97-8.01 (1H, ddd, J=8.2, 7.2, 1.0), 7.80-7.84 (1H, dd, J1=J2=7.6), 7.51 (1H, d, J=2.1), 7.37-7.38 (1H, d, J=2.6), 7.25-7.28 (1H, dd, J=9.4, 2.6), 5.06- 5.08 (1H, t, J=5.2), 5.03-5.06 (1H, t, J=5.3), 4.48 (3H, s), 4.43-4.46 (2H, t, J=4.8), 4.30-4.32 (2H, t, J=4.6), 3.84-3.88 (2H, m), 3.80-3.84 (2H, m).
m/z (ES+): 404.1 (M+)
Example 16 - Preparation of l,3-difluoro-6,10-dimethoxy-13-methylchromenor4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000078_0001
To a suspension of 3,6-dimethoxy-9H-xanthen-9-one (240 mg, 0.937 mmol), 2-bromo-4,6- fluoroaniline (390 mg, 1.87 mmol, 2 equ), Ν,Ν-diisopropylethyl amine (817 μΐ, 4.68 mmol, 5 equ) in toluene (8 mL) was added a 1M solution of titanium tetrachloride in toluene (1.41 mL, 1.41 mmol, 1.5 equ) dropwise, and the resulted suspension was refluxed for 3 hours. The reaction mixture was diluted with dichloromethane and filtered. The filtrate was concentrated to dryness and the solid residue was absorbed on silica and purified by flash chromatography (gradient elution hexane:ethyl acetate 15:1 to 8:1) to give 2-bromo-N-(3,6-dimethoxy-9H- xanthen-9-ylidene)-4,6-difluoroaniline [m/z (ES+): 446 (MH+), 448 (MH+, 81Br)] as a yellow solid (775 mg, 36% yield). The product was used without further characterisation in next step. A mixture of 2-bromo-N-(3,6-dimethoxy-9H-xanthen-9-ylidene)-4,6-difluoroaniline (300 mg, 0.672 mmol), bis(triphenylphosphine)palladium(II) chloride (catalytic amount), 2- Diphenylphosphino-2'-(N,N-dimethylamino)biphenyl (2.5 mg, 0.0673 mmol, 0.1 equ), sodium fert-butoxide (86 mg, 0.896 mmol, 2 equ) and dimethylacetamide (6 mL) was heated at 200°C for 5 minutes under microwave radiation. The reaction mixture was poured into ice and the resulting precipitate was filtered and dried under suction. The solid was absorbed on silica and purified by flash chromatography (gradient elution hexane:ethyl acetate 40:1 to 10:1, then DCM:MeOH 100% to 80%) to give l,3-difluoro-6,10-dimethoxychromeno[4,3,2- g/z]phenanthridine [m/z (ES+): 366 (MH )] as a solid (20 mg, 8% yield). The product was used without further characterisation in next step.
A mixture of l,3-difluoro-6,10-dimethoxychromeno[4,3,2-g i]phenanthridine (20 mg, 0.0548 mmol), methyl trifluoromethansulfonate (60 μί, 0.548 mmol, 10 equ), potassium carbonate (76 mg, 0.548 mmol, 10 equ) and anhydrous chloroform (1 mL) were heated in a sealed tube at 100°C for 72 hours. The reaction mixture was quenched with methanol and filtered; the filtrate was concentrated to dryness and the solid residue was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 100%-95%) to give the title compound as a solid (13 mg, 45% yield).
<¾ (DMSO-Je): 8.66-8.68 (1H, d, J=9.8), 8.58-8.60 (1H, d, J= 9.4), 8.23 (1H, d, J= 2.3), 8.00- 8.06 (1H, ddd, J=11.0, 8.6, 2.6) 7.62-7.63 (1H, d, J=2.2), 7.40-7.41 (1H, d, J=2.7) 7.26-7.29 (1H, dd, J=9.4, 2.6), 4.40-4.43 (3H, d, J=9.6), 4.17 (3H, s), 4.06 (3H, s)
m/z (ES+): 380 (MH+). Example 17 - Preparation of l,ll-difluoro-2-methoxy-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparati -difluoro-4-methoxyphenylboronic acid
Figure imgf000079_0001
Methyl iodide (1.56 mL, 25.1 mmol, 1.5 equ) was added dropwise to a suspension of 4-bromo- 2,3-difluorophenol (3.5 g, 16.7 mmol) and potassium carbonate (2.77 g, 20.0 mmol, 1.2 equ) in N,N-dimethylformamide (30 mL) and the mixture was stirred for 3 days. The precipitate was removed by filtration and the filtrate was concentrated to dryness in vacuo. Dichloromethane and water were added and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate and concentrated in vacuo to give l-bromo-2,3- difluoro-4-methoxybenzene as a white crystalline solid (3.8 g, quantitative).
Note: On a larger scale, brine was added to the reaction mixture and the product was extracted with diethyl ether. The combined organic extracts were washed twice with water, dried over magnesium sulfate and concentrated to dryness in vacuo.
A 1.6M solution of tert-butyl lithium in pentane (23.4 mL, 37.5 mmol, 2.2 equ) was added dropwise under nitrogen at -78°C to a solution of l-bromo-2,3-difluoro-4-methoxybenzene (3.8 g, 17.0 mmol) in anhydrous tetrahydrofuran (40 mL) and the resulting yellow/red solution was stirred for 45 min. at -78 C before the addition of trimethyl borate (5.69 mmol, 51.0 mmol, 3 equ). The resulting pale yellow solution was stirred for 16 h and the reaction was quenched with 3N HC1. The mixture was stirred for 2 h and water was added. The product was extracted with diethyl ether and the combined organic extracts were dried over magnesium sulfate and concentrated in vacuo. The product was triturated with hexane, filtered and dried under suction to give the title compound as an off-white solid (2.53 g, 79% yield).
<¾ (DMSO-d6): 8.16 (2H, s (br)), 7.31-7.36 (1H, ddd, J=8.6, 6.6, 2.3), 6.96-7.00 (1H, ddd, J=8.6, 7.3, 1.5), 3.87 (3H, s).
Note: The estimated purity of the product as determined by NMR was 80% and the product was used without further purification in the next step.
Step 2 - Preparation of 2',3,3'-trifluoro-4'-methoxybiphenyl-2-carbonitrile
Figure imgf000080_0001
Nitrogen was bubbled through a suspension of 2-bromo-6-fhiorobenzonitrile (2.39 g, 12.0 mmol), 2,3-difluoro-4-methoxyphenylboronic acid (2.7 g, 14.4 mmol, 1.2 equ) and potassium fluoride (2.3 g, 39.6 mmol, 3.3 equ) in tetrahydrofuran (45 mL) for 10 min. before the addition of bis(tri-iert-butylphosphine)palladium (123 mg, 0.24 mmol, 0.02 equ). The mixture was immersed into a pre-heating oil bath at 55°C and the mixture was stirred at 55°C for 45 min. The mixture was cooled to room temperature and water was added. The product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate and concentrated in vacuo. The resulting solid was triturated with boiling hexane and the resulting suspension was allowed to cool to room temperature. The precipitate was filtered, washed with hexane and dried under suction to give the title compound as a white solid (2.27 g, 72% yield). <¾ (CDC13): 7.62-7.68 (1H, m), 7.29-7.32 (1H, m), 7.23-7.28 (1H, m), 7.14-7.19 (1H, ddd, J=8.8, 7.7, 2.4), 6.86-6.91 (1H, ddd, J=9.0, 7.4, 1.9), 3.98 (3H, s). Step 3 - Preparation of 3-(2-bromo-4-fluorophenoxy)-2',3'-difluoro-4'-methoxybiphenyl-2- carbonitrile
Figure imgf000081_0001
A suspension of 2',3,3'-trifluoro-4'-methoxybiphenyl-2-carbonitrile (75 mg, 0.285 mmol), 2- bromo-4-fluorophenol (82 mg, 0.427 mmol, 1.5 equ) and potassium carbonate (79 mg, 0.57 mmol, 2 equ) in N,N-dimethylformamide (1.5 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and poured into ice. The resulting fine suspension was transferred into a separating funnel and water and dichloromethane were added. The product was extracted with dichloromethane and the combined organic extracts were washed twice with water, dried over magnesium sulfate and concentrated in vacuo. The resulting off-white solid was triturated with boiling hexane and the suspension was allowed to cool to room temperature. The precipitate was filtered, washed with hexane and dried under suction to give the title compound as an off-white solid (85 mg, 69% yield).
<¼ (CDC13): 7.48-7.52 (1H, dd, J=8.6, 7.8), 7.44-7.46 (1H, dd, J=7.6, 2.8), 7.17-7.25 (3H, m), 7.10-7.15 (1H, m), 6.87-6.91 (1H, ddd, J=9.0, 7.4, 1.9), 6.64-6.66 (1H, dd, J=8.5, 1.1), 3.99 (3H, s,).
m/z (ES+): 456.0 (M+Na+(81Br)), 454.0 (M+Na+(79Br)), 436.0 (MH+(81Br)), 434.0 (MH+(79Br)). Step 4 - Preparation -difluoro-2-methoxychromeno[4,3,2-^A]phenanthridine
Figure imgf000081_0002
A solution of 3-(2-bromo-4-fluorophenoxy)-2',3'-difluoro-4'-methoxybiphenyl-2-carbonitrile (1. 39 g, 3. 20 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise to a solution of fert-butyl lithium (1.6M in pentane, 4 mL, 6.40 mmol, 2 equ) in anhydrous tetrahydrofuran ( 15 mL) at -78°C under nitrogen and the resulting dark red solution was stirred for 5min. at -78°C and was then allowed to warm slowly to room temperature and the mixture was stirred for 30min. The reaction was quenched with acetic acid and 1M NaOH, followed by sat. NaHCC>3 were added. The resulting bright yellow suspension was transferred into a separating funnel and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate. A reaction mixture from a separate experiment carried out on 85 mg scale was combined and the mixture was absorbed on silica. The product was purified by flash chromatography (gradient elution CHCl3:Hexane 25%-50%-100% then DCM:MeOH 97.5%- 95%) to give the title compound as a yellow solid (565 mg, 50% yield).
Notel: the product could be only partially purified as some impurities co-eluted with the product during the purification by flash chromatography. The product was used without further purification in the next steps.
Note2: The pure title compound could be obtained by precipitation in dichloromethane.
<¾ (DMSO-d6): 8.41-8.44 (1H, dd, J=9.2, 1.7), 8.35-8.37 (1H, dd, J=8.4, 0.6), 8.15-8.18 (1H, m), 7.93-7.97 (1H, dd, J=8.2), 7.54-7.58 (3H, m), 7.47-7.49 (1H, dd, J=8.0, 0.6), 4.03 (3H, s).
Step 5 - Preparation of l,ll-difluoro-2-methoxy-13-methylchromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000082_0001
Methyl triflate (98 iL, 0.895 mmol, 10 equ ) was added to a suspension of l,l l-difluoro-2- methoxychromeno[4,3,2-g/z]phenanthridine (30 mg, 0.0895 mmol) and potassium carbonate (124 mg, 0.895 mmol, 10 equ) in chloroform (3 mL) and the mixture was heated at 110°C m a sealed tube overnight. The mixture was cooled to room temperature and the reaction was quenched with methanol. The mixture was stirred for ~30min. and was filtered. The filtrate was absorbed on silica and the product was purified by flash chromatography (gradient elution DCM:MeOH 100%-98%-97%-96%-94%-92%). The resulting solid was triturated with 90% diethyl ethenmethanol, filtered, washed with diethyl ether and dried under suction to give the title compound as an orange solid (8 mg, 18% yield).
<¾ (DMSO-d6): 8.70-8.73 (2H, m), 8.58-8.61 (1H, dd, J=9.6, 2.7), 8.41-8.45 (1H, dd, J1=J2=8.2), 8.01-8.10 (2H, m), 7.97-7.99 9 (1H, d, J=8.1), 7.82-7.87 (1H, dd, J=8.9, 7.6), 4.59- 4.61 (3H, d, J=9.7), 4.12 (3H, s).
m/z (ES+): 350.1 (M+) Example 18 - Preparation of l,ll-difluoro-13-methyl-2-(lH-pyrazol-4-Yl)chromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of l,ll-difluorochromeno[4,3,2-^]phenanthridin-2-yl
trifluoromethanesulfonate
Figure imgf000083_0001
A mixture of l,l l-difluoro-2-methoxychromeno[4,3,2-g i]phenanthridine (578 mg, 1.72 mmol, Example 17) and pyridine hydrochloride (~5g) was heated at 210°C under MW radiation (300W, 200psi, cooling) for lOmin. Water was added to the cooled reaction mixture and the precipitate was filtered, washed with water and dried under suction to give l,l l-difluorochromeno[4,3,2- g/z]phenanthridin-2-ol hydrochloride (588 mg) as an orange solid. The product was then dried in a vacuum oven and was used without further characterization and purification in the next step. A mixture of l,l l-difluorochromeno[4,3,2-g z]phenanthridin-2-ol hydrochloride (588 mg, 1.64 mmol), N-phenyltriflimide (1.17 g, 3.29 mmol, 2 equ) and N,N diisopropylethylamine 85ΊμL, 4. 92 mmol, 3 equ) in 1 : 1 CHC^THF (10 mL) was heated at 55°C overnight. The reaction mixture was transferred into a separating funnel and IN HC1 was added. The product was extracted with dichloromethane and the combined organic extracts were washed with sat. aHC03 then dried over magnesium sulfate and the product was absorbed on silica. The product was purified by flash chromatography (gradient elution Hexane:DCM 90%-80%-70%-50% then 60%.) to give the title compound as a yellow solid (380 mg, 49% yield over 2 steps).
<¾ (CDC13): 8.38-8.41 (1H, ddd, J=8.7, 2.8, 0.7), 8.20-8.22 (1H, dd, J=9.2, 2.0), 8.12-8.14 (1H, dd, J=8.3, 0.7), 7.89-7.93 (1H, dd, J1=J2=8.1), 7.47-7.51 (1H, dd, J=9.2, 6.8), 7.47-7.49 (1H, dd, J=8.1, 0.8), 7.33-7.34 (1H, dd, J=4.5, 0.7), 7.31-7.33 (1H, dd, J=7.2, 2.9),
Step 2 - Preparation of l,ll-difluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2- g/i]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000083_0002
Methyl triflate (532 μί, 4.85 mmol, 10 equ) was added to a suspension of 1,11- difluorochromeno[4,3,2-g¾phenanthridin-2-yl trifluoromethanesulfonate (220 mg, 0.485 mmol) and potassium carbonate (670 mg, 4.85 mmol, 10 equ) in chloroform (5 mL) and the mixture was heated in a sealed tube at 100°C overnight. The mixture was cooled to room temperature and methanol was added. The precipitate was removed by filtration and the filtrate was absorbed on silica. The product was purified by flash chromatography (gradient elution DCM:MeOH 100%- 98%-97%-96%-92%) to give l,l l-difluoro-13-methyl-2-
(trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate (90 mg) as pale yellow solid. The product was only partially purified and was used without further purification in the next step.
A mixture of l,l l-difluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2- g/z]phenanthridin-13-ium (40 mg, 0.085 mmol), lH-pyrazole-4-boronic acid pinacol ester (25 mg, 0.128 mmol, 1.5 equ), tetrakis(triphenylphosphine)palladium(0) (9.8 mg , 0.0085 mmol, 0.1 equ) and sodium acetate (15 mg, 0.187, 2.2 equ) in 3: 1 DME:H20 (1.5 mL) was heated at 90°C under MW radiation (30s. ramp time, 300W, 200psi, cooling system on) for 25 min. 0.5 equ of boronic ester, 0.05 equ of tetrakis(triphenylphosphine)palladium(0) and 2.2 equ of sodium acetate were added and the mixture was re-heated under MW radiation at 90°C for 20 min. (30s. ramp time, 300W, 200psi, cooling system on). A reaction mixture of a separate experiment carried out on 25 mg scale was combined and the mixture was concentrated to dryness in vacuo. The resulting solid was dissolved in DCMrMeOH, absorbed on silica and purified by flash chromatography (gradient elution DCMrMeOH 100%-98%-97%-96%-94%-92%-90%) to give 26 mg of an orange solid. The material was triturated with 3N HCl, filtered, washed successively with water and diethyl ether, then dried under suction to give the title compound as an orange solid (15 mg, 24% yield).
<¾ (DMSO-i¾): 8.78-8.80 (1H, d, J=7.8), 8.71-8.73 (1H, d, J=8.7), 8.62-8.65 (1H, dd, J=9.8, 2.6), 8.44-8.49 (1H, dd, J1=J2=8.2), 8.39 (2H, s (br)), 8.29-8.33 (1H, dd, J=8.5, 6.9), 8.02-8.11 (3H, m), 4.66-4.69 (3H, d, J=10.3).
m/z (ES+): 386.0 (M+) Example 19 - Preparation of 3,9-dimethoxy-13-methylchromeno[4,3,2-gfelphenanthridin- 13-ium trifluoromethanesulfonate
Step 1 - Preparation of 3-(2-bromo-6-methoxyphenoxy)-2'-fluoro-5'-methoxybiphenyl-2- carbonitrile
Figure imgf000085_0001
Potassium carbonate (4.6 g, 0.03 mol, 2 equ) was added to a solution of 2',3-difluoro-5'- methoxybiphenyl-2-carbonitrile (4.0 g, 0.016 mol) and 2-bromo-6-methoxyphenol (5.0 g, 0.024 mol, 1.5 equ) in DMF (100 mL) at 25°C. The reaction mixture was heated at 100°C for 24 hrs, allowed to cool, saturated ammonium chloride (50 mL) added, and the aqueous layer extracted with ether (6 x 100 mL). The extracts were washed with saturated brine (25 mL), dried over sodium sulphate, evaporated to dryness and the residue subjected to flash chromatography (DCM) to give the title compound as a white solid (1.7 g, 24% yield).
<¼ (CDClj): 7.43 (1H, t, J- 8.0), 7.23 (1H, d, J= 8.0), 7.18 - 7.14 (3H, m), 7.03 - 6.96 (3H, m), 6.57 (1H, dd, J= 0.8, 8.4), 3.86 (3H, s), 3.82 (3H, s).
m/z (ES+) : 428.0 (MH+, 79Br) , 430.0 (MH+, 8 xBr) .
Step 2 ine
Figure imgf000085_0002
A solution of 3-(2-bromo-6-methoxyphenoxy)-2'-fluoro-5'-methoxybiphenyl-2-carbonitrile (1.70 g, 3.9 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise to a solution of tert- butyl lithium (5.2 mL (1.6M pentane), 8.3 mmol, 2.1 equ) in anhydrous tetrahydrofuran (40 mL) at -78°C. After 5 mins, the temperature was allowed to warm to 25°C and the reaction stirred for another 30 mins. Saturated ammonium chloride (35 mL) was added and the aqueous layer extracted with dichloromethane (3 x 75 mL). The extracts were dried over sodium sulphate, evaporated to dryness and the residue subjected to flash chromatography (gradient elution DCM:EtOAc 100%-90%) to give the title compound as a white solid (540 mg, 41% yield).
4 (CDClj): 8.22 (1H, m), 8.11 (2H, d, J= 8.0), 7.82 (1H, t, J= 8.0), 7.78 (1H, d, J= 2.4), 7.50 (1H, d, J= 9.4), 7.35 (1H, dd, J= 2.0, 8.0), 7.28 (1H, m), 7.12 (1H, m), 4.05 (3H, s), 4.03 (3H, s). m/z (ES+): 330.0 (MH ).
Step 3 - 3,9-dimethoxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium
tnfluoromethanesulfonate
Figure imgf000086_0001
A suspension of 23,9-dimethoxychromeno[4,3,2-g z]phenanthridine (65 mg, 0.19 mmol) , methyl triflate (162 mg, 1.0 mmol, 5.0 equ) and potassium carbonate (41 mg, 0.29 mmol, 1.5 equ) in chloroform (5 mL) was heated at 130°C in a sealed tube overnight. The inorganics were removed by filtration and washed with 20% methanohDCM (20 mL). The washings were combined with the filtrate, evaporated to dryness and subjected to flash chromatography (gradient elution DCMrMeOH 95%-90%) to give the title compound as a yellow solid (50 mg, 51% yield).
<¾ (DMSO-d6): 8.82 (1H, d, J= 7.6), 8.40 - 8.30 (3H, m), 8.04 (2H, t, J= 8.4), 7.73 - 7.59 (3H, m), 4.61 (3H, s), 4.08 (3H, s).
m/z (ES+): 344.1 (M+). Example 20 - Preparation of 2-(methoxycarbonyl)-13-methylchromeno[4,3<2- gfelphenanthridin-13-ium trifluoromethanesulfonate
- Preparation of methyl chromeno[4,3,2-g¾]phenanthridine-2-carboxylate
Figure imgf000086_0002
A suspension of methyl chromeno[4,3,2-g¾phenanthridin-2-yl trifluoromethanesulfonate (100 mg, 0.2 mmol, Example 4), dichlorobis(triphenylphosphine) palladium(II) (3 mg, 2 mol%) and potassium carbonate (50 mg, 0.36 mmol, 1.5 equ) in anhydrous N,N-dimethylformamide (2 mL) and methanol (1 mL) was heated at 60°C under an atmosphere of carbon monoxide for 3 hrs. The reaction mixture was evaporated to dryness and the residue subjected to flash chromatography (DCM) to give the title compound as a yellow solid (56 mg, 71% yield).
<¾ (CDC13): 8.80 (1H, d, J= 1.6), 8.74 (1H, d, J= 8.0), 8.48 (1H, d, J= 8.4), 8.20 (2H, m), 7.88 (1H, t, J= 8.0), 7.58 (1H, t, J= 8.4), 7.46 (1H, dd, J= 0.8, 8.0), 7.37 (2H, m), 4.04 (3H, s). m/z (ES+): 328.1 (MH ).
Step 2 - Preparation of 2-(methoxycarbonyl)-13-methylchromeno[4,3,2-^A]phenanthridin- 13-ium trifluoromethanesulfonate
Figure imgf000087_0001
A suspension of methyl chromeno[4,3,2-g^]phenanthridine-2-carboxylate (50 mg, 0.15 mmol), methyl triflate (125 mg, 7.6 mmol, 5.0 equ) and potassium carbonate (21 mg, 0.15 mmol, 1.0 equ) in chloroform (5 mL) was heated at 120°C in a sealed tube overnight. The inorganics were removed by filtration and washed with 20% methanol :DCM (15 mL). The washings were combined with the filtrate, evaporated to dryness and subjected to flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a yellow solid (40 mg, 53% yield).
<¼ (DMSO-d6): 9.04 (1H, d, J= 8.4), 8.81 (1H, d, J= 8.0), 8.76 (1H, s), 8.59 (1H, d, J= 8.0), 8.47 (1H, t, J= 8.0), 8.37 (1H, d, J= 8.4), 8.14 (2H, m), 7.95 (1H, d, J= 8.4), 7.76 (1H, t, J= 8.4), 4.70 (3H, s), 4.04 (3H, s).
m/z (ES+): 342.1 (M+).
Example 21- Preparation of 13-methYl-2-(thiophen-3-yl chromeno[4.l3.i2-gfelphenanthridin- 13-ium trifluoromethanesulfonate
Figure imgf000087_0002
13-Methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium
trifluoromethanesulfonate (120 mg, 0.21 mmol, Example 4), potassium carbonate (40 mg, 0.29 mmol, 1.4 equ), thiophen-3-ylboronic acid (34 mg, 0.27 mmol, 1.3 equ), tetrakis triphenylphosphine palladium(O) (11 mg, 5 mol%), 1 ,2-dimethoxyethane (1 mL), and water (0.5 mL) were heated under MW irradiation at 80°C for 15min. (100W, 50psi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (gradient elution DCM:MeOH 5%-90%) to give the title compound as an orange solid (75 mg, 70% yield). <¾ (DMSO-de): 8.95 (IH, d, J= 8.8), 8.78 (1H, d, J= 7.6), 8.58 (2H, m), 8.40 (2H, m), 8.33 (IH, dd, J= 1.6, 8.8), 8.10 (IH, t, J= 7.2), 7.98 (2H, m), 7.93 (IH, dd, J= 0.8, 8.4), 7.83 (IH, dd, J= 2.0, 5.2), 7.74 (IH, t, J= 7.2), 4.73 (3H, s).
m z (ES+): 366.1(M+).
Example 22- Preparation of 2-(3-(dimethylamino)prop-l-vnyl)-13-methylchromenor4,3^2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000088_0001
13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-^¾]phenanthridin-13-ium
trifluoromethanesulfonate (110 mg, 0.19 mmol, Example 4), triethylamine (95 mg, 0. 9 mmol, 5.0 equ), l-dimethylamine-2-propyne (63 mg, 0.7 mmol, 4 equ), tetrakis triphenylphosphine palladium(O) (6 mg, 5 mol%), copper iodide (1 mg, 2 mol%), 1 ,2-dimethoxyethane (1 mL), were heated under MW irradiation at 80°C for 5min. (300W, lOOpsi.). The reaction mixture was then evaporated to dryness, and purified by preparative TLC (20% methanol:DCM) to give the title compound as a yellow solid (15 mg, 15% yield).
<¾ (DMSO-d6): 8.90 (IH, d, J= 8.4), 8.76 (IH, d, J= 7.6), 8.54 (IH, d, J= 7.2), 8.42 (2H, m), 8.04 (IH, t, J= 8.4), 7.96 (IH, d, J= 8.4), 7.92 (2H, m), 7.73 (IH, t, J=7.6), 4.63 (3H, s), 3.61 (2H, s), 2.34 (6H, s).
m/z (ES+): 365.1(M+).
Example 23 - Preparation of 13-methyl-2-(lH-pyrazol-4-yI)chromenor4,3^2- g¾lphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000088_0002
13-Methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/z]phenanthridin-13-ium
trifluoromethanesulfonate (75 mg, 0.13 mmol, Example 4), potassium carbonate (25 mg, 0.18 mmol, 1.4 equ), 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrrole (32 mg, 0.16 mmol, 1.3 equ), tetrakis triphenylphosphine palladium(O) (7 mg, 5 mol%), 1,2-dimethoxyethane (2 mL), and water (1 mL) were heated under MW irradiation at 80°C for 5min. (100W, 50psi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (gradient elution DCM:MeOH 90%-85%) to give the title compound as an orange solid (30 mg, 47% yield).
4 (DMSO-i¾: 13.28 (1H, s (br)), 8.90 (1H, d, J= 8.8), 8.72 (2H, m), 8.68 (1H, d, J= 8.4), 8.41 (1H, s), 8.30 (2H, m), 8.21 (1H, d, J= 1.2), 8.09 (1H, t, J= 8.4), 7.94 (2H, m), 7.73 (1H, t, J= 8.4), 4.70 (3H, s).
m/z (ES+): 350.2 (M+).
Example 24 - Preparation of 2-methoxy-13-methylchromeno[4,3.2-gfelphenanthridin-13-
Figure imgf000089_0001
A suspension of 2-methoxychromeno[4,3,2-g¾phenanthridine (150 mg, 0.5 mmol, Example 4), methyl triflate (410 mg, 2.5 mmol, 5.0 equ) and potassium carbonate (69 mg, 0.5 mmol, 1.0 equ) in chloroform (10 mL) was heated at 120°C in a sealed tube overnight. The inorganics were removed by filtration and washed with 20% methanol :DCM (30 mL). The washings were combined with the filtrate, evaporated to dryness and subjected to flash chromatography (10% methanol:DCM) to give the title compound as a yellow solid (20 mg, 9% yield).
<¾ (DMSO-i¾): 8.87 (1H, d, J= 9.2), 8.65 (1H, d, J= 8.0), 8.54 (1H, d, J= 7.6), 8.37 (1H, t, J= 8.0), 8.07 (1H, t, J= 7.6), 7.90 (2H, d, J= 8.0), 7.60 (2H, m), 7.59 (1H, d, J= 9.2), 4.63 (3H, s), 4.10 (3H, s).
m/z (ES+): 314.0 (M+).
Example 25 - Preparation of 2-(methoxymethvI)-13-methylchromeno[4,3,2- g¾lphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of 2',3-difluoro-4'-formylbiphenyl-2-carbonitrile
Figure imgf000090_0001
Nitrogen was bubbled through a suspension of 2-bromo-6-fluorobenzonitrile (2.2 g, 11 mmol), 2-fluoro-4-formylphenylboronic acid (2.2 g, 13 mmol, 1.2 equ) and potassium fluoride (2.1 g, 36 mmol, 3.3 equ) in tetrahydrofuran (50 mL) for 5min. before the addition of bis(tri-ferf- butylphosphine)palladium (56 mg, 0.11 mmol, 0.01 equ). The mixture was immersed into a preheating oil bath at 50°C and the mixture was stirred at 50°C for 2h. The mixture was cooled to room temperature and brine was added. The product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate and concentrated in vacuo. The resulting solid was triturated with diethyl ether and the resulting precipitate was filtered, washed with hexane and dried under suction to give the title compound as a white solid (2.35 g, 88% yield).
<¾ (CDC13): 10.07-10.06 (1H, d, J= 1.8), 7.83-7.81 (1H, dd, J= 7.8, 1.5), 7.76-7.76 (1H, dd, J= 9.7, 1.4), 7.73-7.67 (1H, m), 7.65-7.61 (1H, m), 7.35-7.30 (2H, m).
m z (ES+); 244.1 (MH+)
Step 2 - Preparation of 3-(2-bromophenoxy)-2'-fluoro-4'-formylbiphenyl-2-carbonitrile
Figure imgf000090_0002
A suspension of 2',3-difluoro-4'-formylbiphenyl-2-carbonitrile (2.2 g, 9.0 mmol), 2-bromophenol (2.34 g, 13.5 mmol, 1.5 equ) and potassium carbonate (2.5 g, 18.1 mmol, 2 equ) in N,N- dimethylformamide (25 mL) was heated at 100°C for 48 h. The mixture was cooled to room temperature and saturated ammonium chloride was added. The product was extracted with diethyl ether (5 x 25 mL) and the combined organic extracts were washed with brine and concentrated in vacuo. The product was purified by flash chromatography (30% EtOAc:Hexane) to give the title compound as a white solid (1.0 g, 28% yield).
<¾ (CDCI3): 10.07-10.07 (1H, d, J=1.8), 7.84-7.81 (1H, dd, J= 7.8, 1.5), 7.77-7.74 (1H, dd, J= 9.5, 1.4), 7.71-7.67 (2H, m), 7.55-7.51 (1H, dd, J= 8.5, 7.8), 7.43-7.38 (1H, ddd, J= 8.2, 7.3, 1.6), 7.22-7.17 (3H, m), 6.74-6.72 (1H, dd, J= 8.6, 0.8). m/z (ES+): 456.0 (M+Na+(81Br)), 454.0 (M+Na+(79Br)), 436.0 (MH+(81Br)), 434.0 (MH+(79Br)), Step 3 - Preparation of 3-(2-bromophenoxy)-2'-fluoro-4'-(hydroxymethyl)biphenyl-2- carbonitrile
Figure imgf000091_0001
Sodium borohydride (9 mg, 2.4 mmol, 2.1 equ) was added at room temperature to a solution of 3-(2-bromophenoxy)-2'-fluoro-4'-formylbiphenyl-2-carbonitrile (440 mg, 1.1 mmol) in methanol (5 mL) and the mixture was stirred for 30min. The mixture was concentrated in vacuo and the product was purified by flash chromatography (10% EtOAc:Hexane) to give the title compound as a white solid (340 mg , 78% yield).
<¾ (CDC13): 7.70-7.68 (1H, dd, J= 7.8, 1.4), 7.52-7.46 (2H, m), 7.41-7.36 (1H, ddd, J= 8.3, 7.3, 1.6), 7.31-7.26 (2H, m), 7.20-7.15 (3H, m), 6.67-6.69 (1H, dd, J=8.0, 0.8), 4.79-4.78 (2H, d, J= 6.0), 1.85-1.82 (1H, t, J- 6.0).
Step 4 - Preparation of 3-(2-bromophenoxy)-2'-fluoro-4'-(methoxymethyl)biphenyl-2- carbonitrile
Figure imgf000091_0002
A solution of 3-(2-bromophenoxy)-2'-fluoro-4'-(hydroxymethyl)biphenyl-2-carbonitrile (370 mg, 0.93 mmol) in anhydrous tetrahydrofuran (2 mL) was added to a suspension of sodium hydride (60% in mineral oil, 46 mg, 1.1 mmol, 1.2 equ) in anhydrous tetrahydrofuran at room temperature and the mixture was stirred for 30min. before the addition of methyl iodide (200 mg, 1.4 mmol, 1.5 equ). The reaction mixture was stirred for 2 h and water was added. The resulting mixture was concentrated to dryness in vacuo and the product was purified by flash chromatography (DCM) to give the title compound as a white solid (330 mg, 86% yield).
4 (CDCI3): 7.70-7.68 (1H, dd, J= 7.9, 1.4), 7.50-7.44 (2H, m), 7.41-7.36 (1H, m), 7.25-7.22 (2H, m), 7.19-7.14 (3H, m), 6.60-6.67 (1H, dd, J= 8.5, 0.8), 4.53 (2H, s), 3.46 (3H, s).
Step 5 - Preparation of 2-(methoxymethyl)chromeno[4,3,2-gft]phenanthridine
Figure imgf000092_0001
A solution of 3-(2-bromophenoxy)-2'-fluoro-4'-(methoxymethyl)biphenyl-2-carbonitrile (320 mg, 0.78 mmol) in anhydrous tetrahydrofuran (2 mL) was added dropwise to a solution of tert- butyl lithium (1.6M in pentane, 1 mL, 1.6 mmol, 2.2 equ) in anhydrous tetrahydrofuran (8 mL) at -78°C under nitrogen and the resulting dark red solution was stirred for 5min. at -78°C and was then allowed to warm slowly to room temperature and the mixture was stirred for 30min. A saturated solution of ammonium chloride was added and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulphate and concentrated in vacuo. The product was purified by flash chromatography (gradient elution DCM:EtOAc 100%-95%) to give the title compound as a yellow solid (100 mg, 41% yield). <¾ (DMSO- ): 8.64-8.62 (1H, d, J= 8.4), 8.59-8.56 (1H, dd, J= 8.0, 1.6), 8.42-8.40 (1H, d, J= 7.8), 7.97-7.93 (2H, m), 7.69-7.64 (1H, ddd, J= 8.6, 7.2, 1.7), 7.58-7.56 (1H, dd, J= 8.3, 1.8), 7.52-7.52 (1H, dd, J= 8.0, 0.7), 7.47-7.45 (1H, dd, J= 8.4, 0.8), 7.44-7.40 (1H, ddd, J= 8.0, 7.2, 1.1), 4.66 (2H, s), 3.39 (3H, s).
m/z (ES+): 314.1 (MH+)
Step 6 - Preparation of 2-(methoxymethyl)-13-methylchromeno[4,3,2-^A]phenanthridin- 13-ium trifluoromethanesulfonate
Figure imgf000092_0002
Methyl triflate (780 mg, 4.8 mmol, 5 equ) was added to a suspension of 2- (methoxymethyl)chromeno[4,3,2-g z]phenanthridine (300 mg, 0.96 mmol) and potassium carbonate (200 mg, 1.4 mmol, 1.5 equ) in chloroform (20 mL) and the mixture was heated at 110°C in a sealed tube overnight. The mixture was cooled to room temperature and the insoluble material were removed by filtration. The filtrate was absorbed on silica and the product was purified by flash chromatography (gradient elution DCM:MeOH 98%-95%) to give the title compound as a yellow solid (130 mg, 30% yield). δΗ (DMSO-d6): 8.93-8.91 (1H, d, J=8.4), 8.76-8.74 (1H, d, J=7.7), 8.59-8.56 (1H, dd, J= 8.5, 1.3), 8.43-8.39 (1H, dd, J= 8.2, 8.2) 8.29 (1H, s), 8.10-8.07 (1H, ddd, J= 8.3, 7.1, 1.3), 8.01-7.99 (1H, dd, 8.4, 0.5), 7.93-7.90 (1H, dd, J= 8.6, 1.1), 7.89-7.87 (1H. d, J= 8.6), 7.73-7.73 (1H, ddd, J= 8.3, 7.2, 1.2), 4.77 (2H, s), 4.64 (3H„ s), 3.46 (3H, s).
m/z (ES+): 328.2 (M+)
Example 26 - Preparation of 2-((dimethylamino)methyl)-13-methyIchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of 2-(bromomethyl)-13-methylchromeno[4,3,2-^]phenanthridin-13- ium trifluoromethanesulfonate
Figure imgf000093_0001
A 1M solution of boron tnbromide in dichloromethane (1.3 mL, 1.3 mmol, 5.0 equ) was added dropwise to a suspension of 2-(methoxymethyl)-13-methylchromeno[4,3,2- z]phenanthridin-
13-ium trifluoromethanesulfonate (130 mg, 0.27 mmol, Example 25) in dichloromethane (5 mL) at 25°C. The resulting mixture was stirred at 25°C overnight, then quenched with a few drops of methanol. After evaporating to dryness, the residue was subjected to flash chromatography (5% methanolrDCM) to give the title compound as a yellow solid (100 mg, 70% yield).
<¾ (DMSO- ): 8.94 (1H, d, J= 8.8), 8.75 (1H, d, J= 7.6), 8.62 (1H, d, J= 8.4), 8.53 (1H, s, J=
1.2), 8.43 (1H, t, J= 8.0), 8.12 (1H, t, J= 7.2), 8.03 (2H, m), 7.93 (1H, dd, J= 1.2, 7.2), 7.71 (1H, t, J= 7.2), 5.06 (2H, s), 4.62 (3H, s).
m/z (ES+): 376.0 (M+, 79Br), 378.0 (M+, 81Br).
Step 2 - Preparation of 2-((dimethylamino)methyl)-13-methylchromeno[4,3,2- g¾]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000093_0002
A 2M solution of dimethylamine in tetrahydrofuran (0.21 mL, 0.4 mmol, 5.0 equ) was added to a solution of 2-(bromomethyl)-13-methylchromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (45 mg, 0.086 mmol) in tetrahydrofuran (3 mL) at 25°C. The resulting solution was stirred for 16 h at 25°C, then evaporated to dryness. The residue was purified by filtering through a short bed of silica (20% methanol :DCM) to give the title compound as an orange solid (15 mg, 36% yield).
<¼ (DMSO-Je): 8.91 (1H, d, J= 8.8), 8.75 (1H, d, J= 8.8), 8.60 (1H, d, J= 8.8), 8.42 (1H, dd, J1=J2= 8.0), 8.20 (1H, m (br)), 8.10 (1H, dd, J1=J2= 7.2), 8.00 (1H, d, J= 8), 7.89-7.94 (2H, m), 7.72 (1H, t, J= 7.6), 4.64 (3H, s), 3.77 (2H, s), 2.40 (6H, s).
m/z (ES+): 341.1 (M+). Example 27 - Preparation of 2-methoxy-13-methylthiochromeno[4,3,2-gfelphenanthridin-
13-ium trifluoromethanesulfonate
Step 1 - Preparation of 3-(2-bromophenylthio)-2'-fluoro-4'-methoxybiphenyl-2-carbonitrile
Figure imgf000094_0001
Potassium carbonate (1.36 g, 9.8 mmol, 2 equ) was added to a solution of 2',3-difluoro-4'- methoxybiphenyl-2-carbonitrile (1.2 g, 5.9 mmol) and 2-bromothiophenol (1.4 g, 7.3 mmol, 2 equ) in N,N-dimethylformamide (12 mL) at 25°C. The reaction mixture was heated at 100°C for 24 hrs, allowed to cool, saturated ammonium chloride (20 mL) added, and the aqueous layer extracted with ether (6 x 50 mL). The extracts were washed with saturated brine (10 mL), dried over sodium sulfate and evaporated to dryness. Recrystallisation from ethyl acetate / heptane gave the title compound as a yellow solid (1.1 g, 54% yield).
<¾ (DMSO-ci6): 7.83 (1H, d, J= 6.4), 7.70 (1H, t, J= 8.0), 7.51 - 7.45 (3H, m), 7.39 - 7.35 (2H, m), 7.27 (1H, dd, J= 1.2, 8.0), 7.04 (1H, dd, J- 2.4, 8.4), 6.95 (1H, dd, J= 2.4, 8.4), 3.86 (3H, s). m/z (ES+): 414.0 (MH+, 79Br), 416.0 (MH+, 81Br).
Step 2 - Preparation of 2-methoxythiochromeno[4,3,2-gA]phenanthridine
Figure imgf000094_0002
A solution of 3-(2-bromophenylthio)-2'-fluoro-4'-methoxybiphenyl-2-carbonitrile (500 mg, 1.2 mmol) in anhydrous tetrahydrofuran (5 mL) was added dropwise to a solution of fert-butyl lithium (1.6 mL (1.6M pentane), 2.5 mmol, 2.1 equ) in anhydrous tetrahydrofuran (15 mL) at - 78°C. After 5 mins, the temperature was allowed to warm to 25°C and the reaction stirred for another 30 mins. Saturated ammonium chloride (20 mL) was added and the aqueous layer extracted with dichloromethane (3 x 50 mL). The extracts were dried over sodium sulfate and evaporated to dryness. Recrystallisation from ethyl acetate / heptane gave the title compound as a yellow solid (150 mg, 40% yield).
<¾ (DMSO-d6): 8.99 (1H, d, J= 6.4), 857 (1H, d, J= 8.8), 8.46 (1H, d, J= 9.2), 7.78 (1H, m), 7.65 (1H, m), 7.54 - 7.50 (4H, m), 7.26 (1H, d, J= 9.2), 3.97 (3H, s).
m/z (ES+): 316.0 (MH+).
Step 3 - Preparation of 2-methoxy-13-methylthiochromeno[4,3,2-^ft]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000095_0001
A suspension of 2-methoxythiochromeno[4,3,2-g/i]phenanthridine (150 mg, 0.47 mmol) , methyl triflate (390 mg, 2.5 mmol, 5.0 equ) and potassium carbonate (66 mg, 0.47 mmol, 1.0 equ) in chloroform (10 mL) was heated at 130°C in a sealed tube overnight. The inorganics were removed by filtration and washed with 20% methanol :DCM (30 mL). The washings were combined with the filtrate, evaporated to dryness and subjected to flash chromatography (10% methanohDCM) to give the title compound as a yellow solid (55 mg, 24% yield).
<¾ (DMSO-ifc): 8.93 (1H, d, J= 9.2), 8.82 (1H, d, J= 7.6), 8.39 (1H, d, J= 9.2), 8.26-8.19 (2H, m), 8.13 (1H, d, J= 7.6), 7.91 (1H, t, J= 8.4), 7.76 (2H, m), 7.62 (1H, dd, J= 2.4, 8.6), 4.53 (3H, s), 4.10 (3H, s).
m/z (ES+): 330.0 (M+).
Example 28 - Preparation of 2-hvdroxy-13-methylthiochronieno[4,3,2-g tlphenanthridin-
Figure imgf000095_0002
A 1M solution of Boron tribromide in dichloromethane (0.42 mL, 0.4 mmol, 5.0 equ) was added dropwise to a suspension of 2-methoxy-13-methylthiochromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (40 mg, 0.08 mmol, Example 27) in dichloromethane (5mL) at 25°C. The resulting mixture was stirred at 25°C overnight, then quenched with a few drops of methanol. After evaporating to dryness, the residue was subjected to flash chromatography (gradient elution DCM:MeOH 95%-93%) to give the title compound as a yellow solid (34 mg, 88% yield).
<¾ (DMSO-d6): 11.22 (1H, s (br)), 8.82 (1H, d, J= 9.2), 8.72 (1H, dd, J= 2.0, 7.2), 8.42 (1H, d, J= 8.0), 8.19-8.01 (3H, m), 7.90 (1H, t, J= 8.4), 7.73 (1H, t, J= 8.4), 7.63 (1H, s), 7.44 (1H, dd, J= 1.6, 8.4), 4.45 (3H, s).
m/z (ES+): 316.1 (M+).
Example 29 - Preparation of 4,ll-difluoro-2-methoxy-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000096_0001
A mixture of 2-bromo-6-fluorobenzonitrile (200 mg, 1 mmol), 2,6-difluoro-4- methoxyphenylboronic acid (224 mg, 1.2 mmol, 1.2 equ), potassium fluoride (196 mg, 3 mmol, 3 equ), bis(triphenylphosphine)palladium (10 mg, 0.0183 mmol, 0.02 equ) and tetrahydrofuran (4 mL) was heated at 120°C for 5 minutes. The reaction mixture was diluted with water and extracted with dichloromethane. The combined organic extracts were concentrated to dryness in vacuo. The resulting solid was absorbed on silica and purified by flash chromatography (gradient elution hexane:ethyl acetate 100:1 to 20:1) giving 2',3,6'-trifluoro-4'-methoxybiphenyl-2- carbonitrile [m/z (ES+): 264 (MH+), 286 (M+Na+)] as a solid (154 mg, 59% yield). The product was used without further characterisation in next step.
A mixture of 2',3,6'-trifluoro-4'-methoxybiphenyl-2-carbonitrile (385 mg, 1.46 mmol), 2-bromo- 4-fluorophenol (420 mg, 2.2 mmol, 1.5 equ), potassium carbonate (403 mg, 2.92 mmol, 2 equ) in N,N-dimethylformamide (8 mL) was heated at 100°C for 6 hours and the mixture was allowed to cool to room temperature. 100 mL of concentrated ammonium chloride were added and the mixture was transferred into a separating funnel. The product was extracted with diethyl ether and the combined organic extracts were washed with brine, dried over magnesium sulfate, filtered and concentrated to dryness in vacuo giving a brown oil that was triturated in hexane. The resulted suspension was filtered giving 3-(2-bromo-4-fluorophenoxy)-2',6'-difluoro-4'- methoxybiphenyl-2-carbonitrile [m/z (ES+): 434 (MH+, 79Br), 436 (MH+, 81Br), 456 (M+Na+, 79Br), 458 (M+Na+, 81Br)] as a solid (430 mg, 68% yield). The product was used without further characterisation in next step.
A solution of 3-(2-bromo-4-fluorophenoxy)-2',6'-difluoro-4'-methoxybiphenyl-2-carbonitrile (430 mg, 0.991 mmol) in anhydrous tetrahydrofuran (5 mL) was added slowly drop wise at - 78°C to a a stirred solution of tert-butyl lithium (1.3 mL, 2.08 mmol, 2.1 equ) in anhydrous tetrahydrofuran (2 mL). The reaction mixture was stirred for 30 minutes at -78°C and was then allowed to warm slowly to room temperature over 18 hours. The reaction mixture was cooled to 0°C and 1 mL of acetic acid was added, then basified to pH 8 by the addition of 1M sodium hydroxide (-15 mL). The mixture was transferred into a separating funnel and the product was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate, filtered and concentrated to dryness in vacuo. The resulting solid was absorbed on silica and purified by flash chromatography (gradient elution hexane:ethyl acetate 1000:1 to 50:1) giving 4,ll-difluoro-2-methoxychromeno[4,3,2-^]phenanthridine [m/z (ES+): 336 (MH+)] as a solid (25 mg, 8% yield). The product was used without further characterisation in next step. A mixture of 4,l l-difluoro-2-methoxychromeno[4,3,2-g/i]phenanthridine (25 mg, 0.0746 mmol), potassium carbonate (18 mg, 0.112 mmol, 1.5 equ), methyl trifluoromethansulfonate (41 μL·, 0.373 mmol, 5equ) and anhydrous chloroform was heated at 120°C in a sealed tube for 18 hours. The reaction mixture was quenched with methanol and concentrated to dryness in vacuo and the resulting solid was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 100%- 10%) to give a solid that was triturated in ether and filtered to give the title compound as a red solid (20 mg, 54% yield).
<¾ (DMSO-d6): 8.62-8.64 (1H, d, J= 8.2), 8.40-8.44 (1H, dd, J= 8.2), 8.32-8.35 (1H, dd, J= 9.9, 2.6), 8.09-8.01 (2H, m), 7.99-8.01 (1H, dd, J= 8.2, 0.5), 7.63-7.67 (1H, dd, J= 14.5, 2.2), 7.61- 7.62 (1H, d, J= 2.2), 4.61 (3H, s), 4.09 (3H, s).
m/z (ES+): 350 (MH+) Example 30 - Preparation of 2-(3-acetamidoprop-l-vnyl)-13-methYlchromeno[4,3<2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000098_0001
A mixture of 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/i]phenanthridin-13- ium (150 mg, 0.25 mmol, Example 4), N-(prop-2-ynyl)acetamide (50 mg, 0.5 mmol, 2 equ), triethylamine (52 mg, 0.5 mmol, 2equ), copper iodide (1 mg, 0.005 mmol, 0.02 equ), tetrakis(triphenylpnosphane)palladium(0) (13 mg, 0.013 mmol, 0.05 equ) and N,N- dimethylformamide (1.5 mL) was heated under microwave radiation at 80°C for 2 minutes. The reaction mixture was concentrated to dryness and the resulting solid was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (DCM:MeOH 90%) to give a sticky orange solid that was stirred at room temperature in diethyl ether for 24 hours. The resulting suspension was filtered to give the title compound as a bright yellow solid (61 mg, 45% yield).
<¾ (DMSO- 6): 8.90-8.92 (1H, d, J=8.7), 8.75-8.76 (1H, d, J=7.8), 8.53-8.55 (1H, dd, J=1.2, 8.4), 8.49-8.52 (1H, t (br), J=5.5), 8.39-8.45 (2H, dd, J=8.2, 16.3), 8.10-8.14 (1H, ddd, J=8.5, 7.2, 1.4), 8.03-8.05 (1H, d, J=8.3), 7.91-7.95 (2H, ddd, 5.0, 3.9, 1.3), 7.71-7.76 (1H, ddd, J=8.4, 7.2, 1.1), 4.62 (3H, s), 4.25-4.26 (2H, d, J=5.5), 1.91 (3H, s).
m/z (ES+): 379 (MH+) Example 31 - Preparation of 2-(dimethylamino)-13-methyIchromenor4,3,2- g/¾1phenanthridin-13-ium iodide
Figure imgf000098_0002
A mixture of 2-acetamido-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (150 mg, 0.3 mmol, Example 8), 6M hydrochloric acid (5 mL, 30 mmol, 100 equ) and methanol (5 mL) was heated at 60°C for 18 hours. The reaction mixture was basified to pH14 with 5M sodium hydroxide and concentrated to dryness in vacuo to give 2- amino-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 73% yield) [m/z (ES+): 299 (MH+)] as a purple solid. The product was used without further characterisation in next step.
A mixture of 2-amino-13-methylchromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.22 mmol), methyl iodide (317 mg, 2.2 mmol, 10 equ) and potassium carbonate (93 mg, 0.67 mmol, 3 equ) in N,N-dimethylformamide (5 mL) were heated in a sealed tube at 100°C for 16 hours. The reaction mixture was concentrated to dryness and the resulting solid was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (DCM:MeOH 95%) to give the title compound as a dark purple solid (60 mg, 59% yield).
4 (DMSO- ): 8.66-8.68 (1H, d, J=9.5), 8.47-8.52 (2H, m), 8.21-8.25, (1H, dd, J=8.2), 8.00- 8.05 (1H, ddd, J=8.5, 7.3, 1.5), 7.83-7.86 (1H, dd, J=8.4, 1.1), 7.69-7.71 (1H, d, J=8.7), 7.64- 7.69 (1H, ddd, J=8.5, 7.3, 1.2), 7.39-7.42 (1H, dd J=9.2, 2.3), 7.10, 7.11 (1H, d, J=2.3), 4.59 (3H, s), 3.24 (6H, s).
m/z (ES+): 327 (MH+).
Example 32 - Preparation of 13-methyl-2-morpholinochromenor4,3,2-g¾lphenanthridin-
13-ium trifluoromethanesulfonate
Figure imgf000099_0001
A mixture of tris(dibenzylideneacetone)dipalladium(0) (8 mg, 0.0085 mmol, 0.05 equ), 2,2'- bis(diphenylphosphino)-l,l'-binaphthyl (11 mg, 0.017 mmol, 0.1 equ) in 1,4-dioxane (2 mL) was heated under microwave radiation at 80°C for 2 minutes. To this was added 13-methyl-2- mo holinochromeno[4,3,2-g ^]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.17 mmol, Example 4), morpholine (75 mg, 0.86 mmol, 5 equ) and cesium carbonate (160 mg, 0.5 mmol, 3 equ) and the mixture was heated under microwave conditions at 115°C for 20 minutes. The reaction mixture was concentrated to dryness and the resulting solid was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (DCM:MeOH 96%) to give the title compound as a dark purple solid (18 mg, 20% yield). <¾ (DMSO- e): 8.72-8.74 (1H, d, J=9.4), 8.55-8.57 (1H, d, J=7.8), 8.50-8.52 (1H, dd, J=8.5, 1.3), 8.26-8.30 (1H, dd, J=8.2), 8.03-8.07 (1H, ddd, J=8.5, 7.2, 1.4), 7.85-7.88 (1H, dd, J=7.5, 1.2), 7.77-7.79 (1H, d, J=7.5), 7.66-7.71 (1H, ddd, J=8.5, 7.4, 1.2), 7.63-7.66 (1H, ddd, J=11.7, 9.4, 2.2), 7.44 (1H, d, J=2.1), 4.60 (3H, s), 3.84-3.87 (4H, t, J=5.1), 3.54 (4H, t, J=5.1).
m/z (ES+): 369 (MH+).
Example 33 - Preparation of ll-fluoro-3-methoxy-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium
Step 1 - ine
Figure imgf000100_0001
A mixture of 2',3-difluoro-5'-methoxybiphenyl-2-carbonitrile (2.49 g, 10.2 mmol), 2-bromo-4- fluorophenol (2.92 g, 15.3 mmol, 1.5 equ) and potassium carbonate (2.82 g, 20.4 mmol, 2 equ) in N,N-dimethylformamide (30 mL) was heated at 100°C in a nitrogen atmosphere for 48 hours. The reaction mixture was allowed to cool to room temperature and saturated ammonium chloride (100 mL) was added. The mixture was transferred into s separating funnel and the product was extracted with diethyl ether. The combined organic extracts were washed with brine, dried over magnesium sulfate, filtered and concentrated to dryness in vacuo. The resulting straw coloured oil was triturated with heptane:EtOAc 4:1 resulting in a suspension that was filtered giving 3-(2- bromo-4-fluorophenoxy)-2'-fluoro-5'-methoxybiphenyl-2-carbonitrile [m/z (ES+): 416 (MH+, 79Br), 418 (MH+, 81Br), 438 (M+Na+, 79Br), 440 (M+Na+, 81Br)] as a white solid. (3.37 g, 80% yield). The product was used without further characterisation in next step.
A solution of 3-(2-bromo-4-fluorophenoxy)-2'-fluoro-5'-methoxybiphenyl-2-carbonitrile (3.37 g, 8.10 mmol) in anhydrous tetrahydrofuran (20 mL) was added slowly drop wise at -78°C to a solution of ierf-butyl lithium (1.6M in pentane, 10.6 mL, 17.0 mmol, 2.1 equ) in anhydrous tetrahydrofuran (50 mL). The reaction mixture was stirred at -78°C for 5 minutes and then at ambient temperature for 30 min. Saturated ammonium chloride (lOOmL) was added and the mixture was extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate and concentrated to dryness in vacuo. The resulting solid was triturated with hexane:EtOAc 4:1 and the suspension was filtered to yield a solid that was purified by flash chromatography (gradient elution hexane hloroform 100%-0%). The product was triturated with hexane:ether 1:1 to give the title compound as a pale orange solid (1.017 g, 40% yield). <¾ (DMSO-de): 8.45-8.47 (1H, d, J= 7.8), 8.14-8.17 (1H, m), 8.04-8.05 (1H, d, J= 2.8), 7.96-7.98 (1H, d, J= 8.9), 7.91-7.95 (1H, dd, J= 8.1) 7.48-7.51 (3H, m), 7.39-7.41 (1H, dd, J= 8.9, 2.8), 3.99 (3H, s)
m z (ES+): 318 (MH+).
Step 2 - Preparation of ll-fluoro-3-methoxy-13-methylchromeno[4,3,2-g i]phenanthridin- 13-ium
Figure imgf000101_0001
A mixture of ll-fluoro-3-methoxychromeno[4,3,2-g z]phenanthridine (100 mg, 0.32 mmol), methyl trifluoromethansulfonate (0.71 mL, 6.3 mmol, 20equ), potassium carbonate (89 mg, 0.64 mmol, 2 equ) in chloroform (10 mL) was heated in a sealed tube at 110°C for 96 hours. The reaction mixture was quenched with methanol and saturated sodium bicarbonate was added, and was then extracted with dichloromethane. The combined organic extracts were dried over magnesium sulfate, filtered and concentrated to dryness in vacuo. The residue was purified by flash chromatography (gradient elution Hexane:DCM 100%-50% to DCM:MeOH 98 -97%- 96%-95%-94%) giving the title compound as a dark orange solid (21 mg, 14% yield).
<¾ (DMSO- ): 8.83-8.85 (1H, d, J=7.9), 8.32-8.42 (4H, m), 7.96-8.00 (3H, m), 7.66-7.69 (1H, dd, J=9.3, 2.7), 4.65 (3H, s), 4.08 (3H, s)
m/z (ES+): 332 (MH+). Example 34 - Preparation of ll-fluoro-13-methyl-3-
(trifluoromethYlsulfonyloxy)chromenor4,3,2-g¾lphenanthridin-13-ium
Step 1 - Preparation of ll-fluorochromeno[4,3,2-g i]phenanthridin-3-yl
trifluoromethanesulfonate
Figure imgf000101_0002
A mixture of l l-fluoro-3-methoxychromeno[4,3,2-g z]phenanthridine (50 mg, 1.58 mmol, Example 33) and pyridine hydrochloride were heated at 210°C under microwave radiation for 5 minutes. The reaction mixture was triturated in water and filtered and the obtained solid washed with hexane giving l l-fluorochromeno[4,3,2-g/z]phenanthridin-3-ol [m/z (ES+): 304 (MH+)] as an orange/brown solid. (440 mg, 92% yield). The product was used without further purification and characterization in the next step.
A mixture of ll-fiuorochromeno[4,3,2-g z]phenanthridin-3-ol (0.44 g, 1.44 mmol), N-Phenyl- bis(trifluoromethanesulfonimide) (1.55 g, 4.33 mmol, 3 equ), Ν,Ν-diisopropylethylamine (560 mg, 4.33 mmol, 3 equ), anhydrous tetrahydrofuran (30 mL) and anhydrous chloroform (30 mL) was heated at 50°C under nitrogen atmosphere for 90 hours. The reaction mixture was concentrated to dryness in vacuo and purified over silica by flash chromatography (gradient elution hexane: chloroform 100%-0%) giving the title compound as a yellow solid (240 mg, 38% yield).
<¼ (DMSO-d6): 8.86 (1H, d, J= 2.8), 8.55-8.57 (1H, d, J= 7.8), 8.21-8.24 (1H, ddd, J= 8.8, 1.7), 8.18-8.20 (1H, d, J= 9.1), 8.03-8.07 (1H, dd, J= 8.2), 7.83-7.86 (1H, dd, J= 9.1, 2.7), 7.63-7.64 (1H, d, J= 7.6), 7.58-7.60 (2H, m)
m/z (ES+): 436 (MH+).
Step 2 - Preparation of ll-fluoro-13-methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2- g¾]phenanthridin- -ium
Figure imgf000102_0001
A mixture of ll-fluorochromeno[4,3,2-g/z]phenanthridin-3-yl trifluoromethanesulfonate (55 mg, 0.126 mmol), methyl trifluoromethansulfonate (451 mg, 2.53 mmol, 20 equ), potassium carbonate (35 mg, 0.252 mmol, 2 equ) and chloroform (10 mL) was heated in a sealed tube at 120°C for 16 hours. The reaction mixture was quenched with methanol and saturated sodium bicarbonate was added. The mixture was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate, filtered and concentrated to dryness. The product was purified by flash chromatography (gradient elution DCM:MeOH 100%-99%-98%- 97%-96%-95%) yielding a brown solid that was triturated in an ethyl acetate:hexane mixture. The yellow suspension was filtered giving the title compound as a solid of the same colour (34 mg, 45% yield).
<¼ (DMSO-if6): 9.18 (1H, d, J=2.8), 8.87-8.89 (1H, d, J=7.8), 8.54-8.56 (1H, d, 9.5), 8.48-8.52 (1H, t, J=8.2), 8.40-8.43 (1H, dd, J=9.7, 2.4), 8.18-8.21 (1H, dd, J=9.5, 2.7), 8.03-8.13 (3H, m), 4.66 (3H, s),
m/z (ES+): 450 (MH+). Example 35 - Preparation of 13-methyl-2-(pyridin-4-yl)chromeno[4,3^2-g¾lphenanthridin- 13-ium methanesulfonate
Figure imgf000103_0001
A mixture of 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/i]phenanthridin-13- ium trifluoromethanesulfonate (110 mg, 0.189 mmol, Example 4), pyridine-4-boronic acid (15.5 mg, 0.227 mmol, 1.2 equ), tetrakis(triphenylphosphane)palladium(0) (1.0 mg, catalytic amount) and potassium carbonate (28.8 mg, 0.378 mmol, 2.0 equ) in l,4-dioxane:H20 1:1 (6 mL) was subjected to the following MW conditions: 80°C, lOmin, 300W, 300psi. The crude reaction mixture was absorbed on silica, then purified by flash chromatography (gradient elution DCM:MeOH 100%-98%-95%-90%-80%) to give the title compound as an orange solid.
The resulting solid (92 mg) was then stirred in DCM:MeOH (20:1, 20 ml) when polymer supported trialkylammonium mesylate resin (2.00 g) was added. After 1 h, the reaction mixture was filtered, concentrated in vacuo and the residue triturated in Et20/hexanes (20 mL) to yield an orange suspension which was collected by filtration and air dried to give a the title compound as orange solid (70 mg, 86% yield).
4 (DMSO-i¾: 9.06-9.10 (IH, d, J=8.7), 8.78-8.85 (3H, m), 8.66 (IH, s), 8.57-8.59 (IH, d, J=8.4), 8.43-8.48 (IH, t, J=8.1), 8.35-8.38 (IH, d, J=8.4), 8.05-8.15 (4H, m), 7.94-7.96 (IH, d, J=8.5), 7.74-7.77 (IH, t, J=7.4), 4.70 (3H, s), 2.30 (3H, s).
m/z (ES+): 361.0 (M+).
Example 36 - Preparation of 4-(13-methylchromeno[4,3,2-gfelphenanthridin-13-ium-2- vDpyridine 1 -oxide methanesulfonate
Figure imgf000103_0002
To a solution of 13-methyl-2-(pyridin-3-yl)chromeno[4,3,2-gA]phenanthridin-13-ium methanesulfonate (20 mg, 0.044 mmol, Example 35) in acetone (10 mL) and MeOH (0.2 mL) at 0°C was added 3,3-dimethyldioxirane (4 mL) in one portion. After 20 mins the reaction mixture was concentrated in vacuo, and the residue triturated in MeOH/Et20 (10 mL) to give the title compound as orange solid (15 mg, 72% yield).
<¼ (DMSO-i¾): 9.00-9.01 (1H, d, J=8.6), 8.79-8.80 (1H, d, J=8.0), 8.60 (1H, s), 8.56-8.58 (1H, d, J=8.0), 8.41-8.45 (3H, m), 8.32-8.34 (1H, d, J=8.5), 8.17-8.19(2H, d, J=7.0), 8.09-8.13 (1H, t, J=7.5), 8.01-8.03 (1H, d, J=8.2), 7.92-7.94 (1H, d, J=8.3), 7.72-7.76 (1H, t, J=8.0), 4.74 (3H, s), 2.30 (3H, s), 2.30 (3H, s).
m/z (ES+): 377.1 (M+).
Example 37 - Preparation of 3,ll-difluoro-13-methylchromenor4,3,2-gMphenanthridin-13- ium trifluoromethanesulfonate
Step 1 - Preparation of 2',3,5'-trifluorobiphenyl-2-carbonitrile
Figure imgf000104_0001
A solution of 2-bromo-6-fluorobenzonitrile (2.00 g, 10.00 mmol), 2,5-difluorophenylboronic acid (1.89 g, 12.00 mmol, 1.2 equ) and potassium fluoride (1.92 g, 33.00 mmol, 3.3 equ) in dry tetrahydrofuran (35 mL). Nitrogen was then bubbled through the reaction mixture for 10 mins when bis(tritertbutylphosphine)palladium (0.10 g, 0.2 mmol, 0.02 equ) was added in one portion then a condenser was fitted, a nitrogen atmosphere applied, and the reaction mixture was then lowered into a pre-heated (50°C) oil bath. After 0.5 h, the reaction was allowed to cool to room temperature when saturated brine solution (50 mL) was added and the reaction mixture was extracted with dichloromethane (5 x 50 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo then dried in vacuo at 40°C to give the title compound as off-white solid (1.727 g, 74% yield).
<¾ (DMSO-d6): 7.89-7.94 (1H, m), 7.63-7.68 (1H, dd, J=8.6,0.8), 7.43-7.55 (4H, m).
m/z (ES+): 234.1 (MH+).
Step 2 - Preparation of 3-(2-bromo-4-fluorophenoxy)-2',5'-difluorobiphenyl-2-carbonitrile
Figure imgf000105_0001
To a solution of 2',3,5'-trifluorobiphenyl-2-carbonitrile (1.00 g, 4.288 mmol) in anhydrous N,N- dimethylformamide (40 mL) under nitrogen was added 2-bromo-4-fluorophenol (1.229 g, 6.433 mmol, 1.5 equ) followed by potassium carbonate (1.185 g, 8.577 mmol, 2.0 equ) in one portion, a condenser was fitted and the reaction mixture was heated to 100°C. After 16h, the reaction was allowed to cool to room temperature then poured into saturated ammonium chloride (50 mL) and extracted with diethyl ether (5 x 25 mL). The combined organic extracts were washed with saturated brine (20 mL), dried over magnesium sulfate, filtered and concentrated in vacuo and air dried to give the title compound as a light brown solid (1.105 g, 64% yield).
δα (DMSO-rf6): 7.84-7.87 (1H, dd, J=8.2,3.0), 7.70-7.74 (1H, dd, J=8.6,7.8), 7.40-7.53 (5H, m), 7.34-7.36 (1H, d, J=7.8), 6.83-6.85 (1H, dd, J=8.6,0.8).
m/z (ES+): 404.0 (MH+, 79Br), 406.0 (MH+, 81Br).
Step 3 - Preparation of 3,ll-difluorochromeno[4,3,2-^]phenanthridine
Figure imgf000105_0002
A 25mL flask was charged with ferf-butyl lithium (1.81 mL, 2.722 mmol, 2.2 equ) and anhydrous tetrahydrofuran (10 mL) under nitrogen at room temperature then cooled to -78°C when a solution of 3-(2-bromo-4-fluorophenoxy)-2',5'-difluoro-5-methylbiphenyl-2-carbonitrile (500 mg, 1.237 mmol, 1 equ) in anhydrous tetrahydrofuran (5 mL) was added dropwise over 2 mins and the reaction mixture was allowed to warm to room temperature. After 2 h the reaction mixture was quenched by the addition of acetic acid (2 mL) then saturated Na2C03 solution (25 mL) was added and the mixture was extracted with dichloromethane (3 x 50 mL). The combined organics were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was then triturated with DCM to give the title compound as a tan brown solid (110 mg, 29% yield) <¾ (DMSO-&): 8.50-8.54 (IH, dd, J=10.4,3.1), 8.45-8.47 (IH, d, J=7.7), 8.21-8.22 (IH, t, J=1.2), 8.19-8.20 (IH, dd, J=2.4,1.2), 8.08-8.12 (IH, dd, J=9.0,5.6), 7.97-8.01 (IH, t, J=8.1), 7.63-7.68 (IH, dt, J=8.8,2.8), 7.53-7.58 (3H, m).
m z (ES+): 306.1 (MH+).
Step 4 - Preparation of 3,ll-difluoro-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000106_0001
A sealed tube was charged with 3,ll-difluorochromeno[4,3,2-g/z]phenanthridine (30 mg, 0.223 mmol) and diluted with chloroform (5 mL) when methyl trifluoromethanesulfonate (0.11 mL, 2.227 mmol, 10 equ) was added in one portion followed by the addition of potassium carbonate (137 mg, 2.227 mmol, 10 equ) and the reaction was heated to 120°C. After 24 h a further amount of methyl trifluoromethanesulfonate (0.11 mL, 2.227 mmol, 10 equ) was added in one portion and after a further 4 h, the reaction mixture was cooled to room temperature and quenched by the slow addition of methanol, filtered and concentrated in vacuo. The crude reaction mixture was absorbed on silica, then purified by flash chromatography (gradient elution DCM:MeOH 100%- 99%-98%-97%-96%-95%-94%-93%-92%-91%-90%) to give recovered starting material and the title compound as an orange solid (22 mg, 75% yield corrected for recovered starting material). 4 (DMSO-d6): 8.84-8.87 (IH, dd, J=9.7,2.9), 8.77-8.79 (IH, d, J=7.6), 8.39-8.49 (3H, m), 7.95- 8.07 (4H, m), 4.66 (3H, s).
m/z (ES+): 320.0 (M+).
Example 38 - Preparation of l,12-difluoro-13-methylchromenor4,3,2-gfelphenanthridin-13- ium trifluoromethanesulfonate
Step 1 - Preparation of 2',3,3'-trifluorobiphenyl-2-carbonitrile
Figure imgf000106_0002
A solution of 2-bromo-6-fluorobenzonitrile (528 mg, 2.64 mmol, 1 equ), 2,3- difluorophenylboronic acid (50 mg, 3.17 mmol, 1.2 equ) and potassium fluoride (506 mg, 8.71 mmol, 3.3 equ) in dry tetrahydrofuran (20 mL). Nitrogen was then bubbled through the reaction mixture for 10 mins when bis(tritertbutylphosphine)palladium (27 mg, 0.05 mmol, 0.02 equ) was added in one portion when a condenser was fitted, a N2 atmosphere applied, and the reaction mixture was then lowered into a pre-heated (50°C) oil bath. After 2 h, the reaction was heated to 75°C. After 24 h, a further amount of bis(tritertbutylphosphine)palladium (27 mg, 0.05 mmol, 0.02 equ) was added and the reaction mixture stirred at 50°C. After a further 48h, the reaction mixture was allowed to cool to room temperature when saturated brine solution (50 mL) was added and the reaction mixture was extracted with dichloromethane (3 x 100 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo then triturated with hexanes to give the title compound as cream coloured solid. The remaining filtrate was concentrated in vacuo and then purified by flash chromatography (gradient elution DCM:Hexanes 100%-0%) to give additional title compound as a white solid (232 mg, 38% yield).
m/z (ES+): 234.1 (MH+).
Step 2 - Preparation of l,12-difluorochromeno[4,3,2-gA]phenanthridine
Figure imgf000107_0001
To a solution of 2',3,3'-trifluorobiphenyl-2-carbonitrile (180 mg, 0.771 mmol, 1 equ) in anhydrous N,N-dimethylformamide (6 mL) under nitrogen was added 2-bromo-3-fluorophenol (0.125 mL, 1.12 mmol, 1.5 equ) followed by potassium carbonate (213 mg, 1.54 mmol, 2.0 equ) in one portion, a condenser was fitted and the reaction mixture was heated to 100°C. After 24 h, the reaction was allowed to cool to room temperature then poured into saturated ammonium chloride (100 mL) and extracted with Et20 (5 x 100 mL). The combined organic extracts were washed with saturated brine (50 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. The material was then triturated with Hexanes:Et20 to give 3-(2-bromo-3- fluorophenoxy)-2',3'-difluorobiphenyl-2-carbonitrile as a light brown solid (m/z (ES+): 404.0 (MH+, 79Br), 406.0 (MH+, 81Br)) (0.167 g, 54% yield). The product was used without further characterization in the next step.
A 25 mL flask was charged with anhydrous tetrahydrofuran (5 mL) under N2 then cooled to - 78°C when a solution of /erf-butyl lithium (0.543 mL, 0.868 mmol, 2.1 equ) was added. After 5 mins, a solution of 3-(2-bromo-3-fluorophenoxy)-2',3'-difluorobiphenyl-2-carbonitrile (167 mg, 0.413 mmol, 1 equ) in anhydrous tetrahydrofuran (5 mL) was added dropwise over 2 mins and the reaction mixture was allowed to warm to room temperature. After 0.5 h the reaction mixture was quenched by the addition of saturated ammonium chloride (100 mL) and the mixture was extracted with dichloromethane (3 x 50 mL). The combined organics were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was then triturated with Hexanes:Et20 to give the title compound as a green solid (43 mg, 34% yield).
<¼ (DMSO-i¾): 8.37-8.54 (2H, m), 8.00-8.02 (1H, m), 7.54-7.74 (3H, m), 7.31-7.35 (1H, m), 7.15-7.22 (1H, m), 6.92-7.03 (1H, m).
m/z (ES+): 306.0 (MH+).
Step 3 - Preparation of l,12-difluoro-13-methylchromeno[4,3,2-^]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000108_0001
A sealed tube was charged with l,12-difluorochromeno[4,3,2-g/z]phenanthridine (20 mg, 0.066 mmol, 1.0 equ) and diluted with chloroform (10 mL) when methyl trifluoromethanesulfonate (0.14 mL, 1.131 mmol, 20 equ) was added in one portion followed by the addition of potassium carbonate (18 mg, 0.132 mmol, 2 equ) and the reaction was heated to 120°C. After 16 h the reaction mixture was cooled to room temperature and quenched by the slow addition of methanol, then saturated NaHCC>3 and the product was extracted with dichloromethane (3 x 10 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. The crude reaction mixture was absorbed on silica, then purified by flash chromatography (gradient elution DCM:MeOH 100%-98%-97%-95%) to give the title compound as a light brown solid (13 mg, 75% yield).
4 (DMSO-d6): 8.79-8.81 (1H, d, J=7.8), 8.76-8.78 (1H, m), 8.47-8.51 (1H, t, J=8.2), 8.19-8.24 (1H, dt, J=8.4,6.2), 8.10-8.12 (1H, d, J=8.2), 7.95-7.98 (2H, m), 7.83-7.85 (1H, d, J=8.6), 7.62- 7.68 (1H, ddd, J=l 1.8,8.3,0.8), 4.31-4.35 (3H, t, J=9.3).
m/z (ES+): 320.1 (M+). Example 39 - Preparation of l-fluoro-2-methoxy-13-methyl-10-
(trifluoromethoxy)chromeno[4,3,2-gfelphenanthridin-13-ium trifluoromethanesulfonate Step 1 - Preparation of 3-(2-bromo-5-(trifluoromethoxy)phenoxy)-2',3'-difluoro-4'- methoxybiphenyl-2-carbonitrile
Figure imgf000109_0001
To a solution of 2\3,3'-trifluoro-4'-methoxybiphenyl-2-carbonitrile (250 mg, 0.950 mmol, Example 17) in anhydrous N,N-dimethylformamide (6 mL) under nitrogen was added 2-bromo- 5-(trifluoromethoxy)phenol (0.209 mL, 1.43 mmol, 1.5 equ) followed by potassium carbonate (263 mg, 1.90 mmol, 2.0 equ) in one portion, a condenser was fitted and the reaction mixture was heated to 100°C. After 96 h, the reaction was allowed to cool to room temperature then poured into saturated ammonium chloride (100 mL) and extracted with diethyl ether (3 x 100 mL). The combined organic extracts were washed with saturated brine (50 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. The material was subjected to hot filtration in hexanes and the filtrate was cooled whereupon precipitation of a solid occurred. This was collected by filtration and air dried to give the title compound as a white solid (180 mg, 38% yield).
m/z (ES+): 499.9 (MH+, 79Br), 501.9 (MH+, 81Br).
Step 2 - Preparation of l-fluoro-2-methoxy-10-(trifluoromethoxy)chromeno[4,3,2- gh ] phenanthridine
Figure imgf000109_0002
A 25mL flask was charged with anhydrous tetrahydrofuran (5 mL) under nitrogen then cooled to -78°C when a solution of ferf -butyl lithium (0.472 mL, 0.757 mmol, 2.1 equ) was added. After 5 mins, a solution of 3-(2-bromo-5-(trifluoromethoxy)phenoxy)-2',3'-difluoro-4'-methoxybiphenyl- 2-carbonitrile (180 mg, 0.360 mmol, 1 equ) in anhydrous tetrahydrofuran (10 mL) was added dropwise over 2 mins and the reaction mixture was allowed to warm to room temperature. After 16 h the reaction mixture was quenched by the addition of saturated ammonium chloride (100 mL) and the mixture was extracted with dichloromethane (3 x 50 mL). The combined organics were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was then triturated with Hexanes to give the title compound as a pale yellow solid (34 mg, 24% yield). m/z (ES+): 402.1 (MH+).
Step 3 - Preparation of l-fluoro-2-methoxy-13-methyl-10-
(trifluoromethoxy)chromeno[4,3,2-g¾]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000110_0001
A sealed tube was charged with 1 -fluoro-2-methoxy-10-(trifluoromethoxy)chromeno [4,3,2- g/z]phenanthridine (34 mg, 0.085 mmol, 1.0 equ) and diluted with chloroform (10 mL) when methyl trifluoromethanesulfonate (0.19 mL, 1.69 mmol, 20 equ) was added in one portion followed by the addition of potassium carbonate (24 mg, 0.17 mmol, 2 equ) and the reaction was heated to 120°C. After 16 h the reaction mixture was cooled to room temperature and quenched by the slow addition of methanol, then saturated NaHC(¾ and the product was extracted with dichloromethane (3 x 10 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. The crude reaction mixture was absorbed on silica, then purified by flash chromatography (gradient elution DCM:MeOH 100%-99%-98%-97%-96%- 95%) to give the title compound as an orange solid (23 mg, 65% yield).
<¾ (DMSO-ife): 8.88-8.90 (1H, d, J=9.3), 8.72-8.74 (2H, d, J=8.1), 8.41-8.45 (1H, t, J=8.2), 8.02- 8.04 (1H, d, J=1.4), 7.96-7.98 (1H, d, J=8.0), 7.84-7.88 (1H, dd, J=9.1,7.9), 7.65-7.67 (1H, d, J=9.3), 4.56-4.59 (3H, d, J=9.9), 4.12 (3H, s).
m/z (ES+): 416.1 (M+).
Example 40 - Preparation of 3-methoxy-13-methylchromeno[4,3^2-gfelphenanthridin-13- ium trifluoromethanesulfonate
Step 1 - Preparation of 2',3-difluoro-5'-methoxybiphenyl-2-carbonitrile
Figure imgf000110_0002
2-Bromo-6-fluorobenzonitrile (4.9 g, 24.52 mmol), 2-fluoro-5-methoxybenzeneboronic acid (5 g, 29.42 mmol, 1.2 equ) and potassium fluoride (4.79 g, 80.92 mmol, 3.3 equ) were stirred in anhydrous tetrahydrofuran (100 mL) at room temperature. Nitrogen was bubbled through for 10 mins before bis-(tri-iert-butylphosphine)palladium (0) (251 mg, 0.490 mmol, 0.02 equ) was added, the solution turned from a light orange colour to a charcoal grey slurry after 10 mins. The flask was sealed and heated to 50°C for 2 h. The mixture was then cooled and partitioned between brine (50 mL) and dichloromethane (50 mL) and the aqueous phase extracted with dichloromethane (3 x 100 mL). The organic extracts were combined, dried over magnesium sulfate and concentrated to dryness in vacuo yielding a light brown solid. The residue was then heated to reflux in hexane (100 mL) and filtered hot. The white precipitate, which formed upon concentration of the filtrate was filtered and dried under suction to give the title compound as white solid (5.32 g, 88% yield).
4 (DMSO-i¾): 7.86-7.92 (1H, ddd, J= 16.3, 6.1, 0.58), 7.60-7.64 (1H, ddd, J= 9.3, 8.6, 0.8), 7.49-7.51 (1H, d, J= 7.7), 7.33-7.37 (1H, ddd, J= 10.1, 9, 0.9), 7.09-7.13 (2H, m), 3.80 (3H, s). m/z (ES+) : 246.1 (M+H+) .
Step 2 - Preparati -(2-bromophenoxy)-2'-fluoro-5'-methoxybiphenyl-2-carbonitrile
Figure imgf000111_0001
2-Bromophenol (5.63 g, 32.54 mmol, 1.5 equ), 2',3-difluoro-5'-methoxybiphenyl-2-carbonitrile (5.32 g, 21.69 mmol) and freshly ground potassium carbonate (6.0 g, 43.88 mmol, 2 equ) were stirred in N,N-dimethylformamide (30 mL) at 100°C under nitrogen for 24 hours. The reaction was cooled to room temperature and a saturated solution of ammonium chloride (100 mL) was added. The product was extracted with diethyl ether (3 x 100 mL) and the combined organic extracts were washed with brine (50 mL), dried over magnesium sulphate and concentrated in vacuo to yield an orange oil. The product was precipitated in heptane: EtOAc 4:1, filtered and dried under suction to give the title compound as a white solid (5.69 g, 66% yield).
<¾ (DMSO-i¾):7.83-7.85 (1H, dd, J= 8, 1.5), 7.69-7.73 (1H, dd, J= 8.5, 0.7), 7.51-7.55 (1H, ddd, J= 9.6, 8.1, 1.5), 7.29-7.40 (4H, m), 7.09-7.12 (2H, m), 6.79-6.81 (1H, dd, J= 8.5, 0.8), 3.81 (3H, s).
m/z (ES+): 398.0 (M+H+, 79Br), 400.0 (M+H+, 81Br).
Step 3 - Preparation of 3-methoxychromeno[4,3,2-g/i]phenanthridine
Figure imgf000112_0001
Tert-butyl lithium (18.75 mL, 1.6M in pentane, 30 mmol, 2 equ) was added to anhydrous tetrahydrofuran (80 mL) at -78°C and the mixture was stirred for 5 mins. before the dropwise addition of a solution of 3-(2-bromophenoxy)-2'-fluoro-5'-methoxybiphenyl-2-carbonitrile (5.69 g, 14.3 mmol) in anhydrous tetrahydrofuran (20 mL). The resulting deep red solution was stirred for 5 min. at -78°C and was then stirred at room temperature for 30 mins. A saturated solution of ammonium chloride (100 mL) was added and the mixture extracted with dichloromethane (3 x 50 mL). The combined organic extracts were dried over sodium sulfate and concentrated. The resulting yellow solid was triturated with diethyl ether and the precipitate was filtered and dried under suction. The mixture was adsorbed onto silica and purified by flash silica chromatography (CHCI3) to give the title compound as a white solid (3.10 g, 73% yield).
4 (DMSO-J6): 8.52-8.54 (1H, dd, J= 7.9, 1.6), 8.44-8.46 (1H, d, J= 7.9) 8.04-8.05 (1H, d, J= 2.7), 7.95-7.98 (1H, d, J= 8.9), 7.90-7.94 (1H, dd, Ji=J2= 8.1), 7.59-7.64(lH, ddd, J= 8.5, 7.1, 1.7), 7.48-7.50 (1H, d, J= 7.5), 7.37-7.51 (3H, m), 4.00 (3H, s).
m/z (ES+): 300.1 (M+H+).
Step 4 - Preparation of 3-methoxy-13-methylchromeno[4,3,2-^]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000112_0002
A suspension of 3-methoxychromeno[4,3,2-g/z]phenanthridine (100 mg, 0.334 mmol, 1 equ), potassium carbonate (93 mg, 0.668 mmol, 2 equ) and methyl trifluoromethanesulfonate (0.756 mL, 6.68 mmol, 20 equ) in chloroform (10 mL) was heated at 110°C in a sealed tube for 72 hours. The reaction was cooled and quenched with methanol. The solution was transferred to a flask where a saturated solution of NaHCC>3 was added slowly with stirring. The product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo. The resulting yellow solid was triturated with diethyl ether, filtered and air dried to yield a yellow solid. This was then adsorbed onto silica and purified by flash silica chromatography (gradient elution, hexane:EtOAc 100%-90% to DCM:MeOH 100%-98%-97%-96%-95%-90%) to give the title compound as a yellow solid (92 mg, 60% yield).
4 (DMSO-i¾: 8.81-8.83 (1H, d, J= 7.7), 8.53-8.55 (1H, dd, 8.1, 1.2), 8.31-8.41 (3H, m), 8.03- 8.07 (1H, ddd, J= 9.8, 7.2, 1.3), 7.97-8.00 (1H, dd, J= 8.1, 0.5), 7.87-7.89 (1H, dd, J= 8.5, 1.1), 7.65-7.71 (2H, m), 4.63 (3H, s), 4.08 (3H, s).
m/z (ES+): 314.1 (M+).
Example 41 - Preparation of 13-methyl-3-(trifluoromethylsulfonyloxy)chromenor4,3.i2- g/¾lphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparat methanesulfonate
Figure imgf000113_0001
3-Methoxychromeno[4,3,2-g¾]phenanthridine (3.00 g, 10.02 mmol, Example 40) were stirred with pyridine.HCl (12 g, 108 mmol, 10 equ) in the MW at 210°C for 5 mins in 6 equal vials (200W, lOOpsi with cooling system on). The vials were then warmed, water was added and the precipitate was filtered, washed with water to give chromeno[4,3,2-g/z]phenanthridin-3-ol as an orange/ brown solid (3.06 g, quantitative yield) which contained unknown impurities. The product was used without further purification in the next step.
A solution of chromeno[4,3,2-g/i]phenanthridin-3-ol (3.06 g, 10.73 mmol, 1 equ), N- phenyltriflimide (11.50 g, 32.18 mmol, 3 equ) and N,N-diisopropylethylamine (4.15 g, 32.15 mmol, 3 equ) in 1:1 chloroform:tetrahydrofuran (100 mL) was stirred at 50°C for 72 hours under nitrogen. The reaction was cooled to room temperature and adsorbed onto silica. The mixture was purified by flash silica chromatography (gradient elution, CHCl3:hexane 50%-100%) to give the title compound as a cream coloured solid (2.98 g, 67% yield).
<¼ (DMSO- ): 8.84-8.85 (1H, d, J= 3.1), 8.58-8.60 (1H, dd, J- 6.5, 1.5), 8.53-8.55 (1H, d, J= 7.7), 8.16-8.19 (1H, d, J= 9.1), 8.02-8.06 (1H, dd, J= 8.1), 7.81-7.84 (1H, dd, J= 6.2, 2.7), 7.69- 7.73 (1H, ddd, J= 8.7, 7.1, 1.7), 7.62-7.64 (1H, dd, J= 7.4, 0.6), 7.47-7.50 (1H, dd, J= 7.6, 0.7), 7.43-7.45 (1H, ddd, J- 9.03, 7.2, 1.1).
m/z (ES+): 418 (M+H+).
Step 2 - Preparation of 13-methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2- g^]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000114_0001
A suspension of chromeno[4,3,2-g z]phenanthridin-3-yl trifluoromethanesulfonate (500 mg, 1.20 mmol), potassium carbonate (334 mg, 2.40 mmol, 2 equ) and methyl trifluoromethanesulfonate (2.71 mL, 23.96 mmol, 20 equ) in chloroform (50 mL) was stirred at 110°C overnight. The reaction was cooled to room temperature and the reaction was quenched with methanol and was transferred to a flask where a saturated solution of NaHCC>3 was added slowly with stirring. The product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulfate, filtered and concentrated to give a yellow solid. The solid was adsorbed onto silica and purified by flash silica chromatography (gradient elution DCM:MeOH 100%-99%- 98%-97%-96%-95%-94 -93%-92%-91%-90%) to give the title compound as a yellow solid (620 mg, 93% yield).
<¼ (DMSO- ): 9.17 (1H, d, J= 2.8), 8.66-8.88 (1H, d, J= 7.6), 8.46-8.50 (1H, ddd, 8.4, 7.9, 0.5), 8.55-8.57 (1H, d, J= 9.5), 8.57-8.59 (1H, dd, J=8.4, 1.2), 8.17-8.20 (1H, dd, J= 9.4, 2.7), 8.10- 8.15 (2H, m), 7.94-7.97 (1H, dd, J= 8.5, 1.1), 7.72-7.76 (1H, ddd, J= 8.3, 7.1, 1.1), 4.63 (3H, s). m/z (ES+): 432.0 (M +).
Example 42 - Preparation of 13-methyl-3-(lH-pyrazol-4-yI)chromenor4,3^2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000114_0002
Nitrogen was bubbled through a mixture of 13-methyl-3- (trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.172 mmol, Example 41), tetramethyldioxaborolanpyrazole (47 mg, 0.241 mmol, 1.4 equ) and potassium carbonate (32 mg, 0.224 mmol, 1.3 equ) in 4:1 DME: Ι¾0 (2.5 mL) at room temperature in a microwave vial. tetrakis(triphenylphosphine)palladium(0) (10 mg, 0.009 mmol, 0.05 equ) was added and the mixture was heated under MW radiation at 80°C for 30 mins (200 W, 200 psi). The mixture was adsorbed onto silica and purified by flash silica chromatography (gradient elution DCM:MeOH 100%-99%-98%-97%-96%-95%-94%-93%-92%-91 %-90%) to give the title compound as a yellow solid (6 mg, 7% yield).
4 (DMSO-de): 13.23 (1H, s), 9.09 (1H, s), 8.95-8.97 (1H, d, J= 8.2), 8.68-8.69 (1H, m), 8.56- 8.58 (1H, d, J= 7.9), 8.32-8.46 (4H, m), 8.06-8.10 (1H, ddd, J= 9.5, 7.2, 1.7), 8.00-8.02 (1H, dd, J= 8.3, 0.68), 7.90-7.92 (1H, d, J= 8), 7.68-7.73 (1H, m), 4.65 (3H, s).
m/z (ES+): 350.1 (M +).
Example 43 - Preparation of 13-methyl-3-(thiophen-3-yl)chromeno[4,3.i2- g/¾1phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000115_0001
13-Methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium
trifluoromethanesulfonate (100 mg, 0.172 mmol, Example 41), thiophene-3-boronic acid (31 mg, 0.241 mmol, 1.4 equ), potassium carbonate (32 mg, 0.224 mmol, 1.3 equ) were stirred in 4: 1 DME:H20 (2.5 mL) at room temperature under nitrogen in a microwave vial. tetrakis(triphenylphosphine)palladium(0) (10 mg, 0.009 mmol, 0.05 equ) was added and nitrogen was bubbled through the mixture before heated at 80°C under microwave radiation for 15 mins (200 W, 200 psi). The mixture was then purified by flash silica chromatography (gradient elution DCM:MeOH 100 -99%-98%-97%-96%-95 -94%-93%-92%-91%-90 ) to give the title compound as a pale orange solid (52 mg, 58% yield).
<¾ (DMSO- ): 9.19 (1H, d, J= 1.5), 9.01-9.03 (1H, dd, J= 8.4, 0.6), 8.57-8.59 (1H, dd, J= 8.4, 1.25), 8.41-8.47 (4H, m), 8.07-8.11 (1H, ddd, J= 8.4, 7.1, 1.3), 8.01-8.04 (2H, m), 7.90-7.93 (1H, dd, J= 8.5, 1.1), 7.80-7.82 (1H, dd, J= 5, 2.9), 7.71-7.73 (1H, ddd, J= 8.3, 7.2, 1.6), 4.66 (3H, s). m/z (ES+): 366.1 (M +). Example 44 - Preparation of 3-(3-(dimethylamino)prop-l-vnyl)-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000116_0001
13-Methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium
trifluoromethanesulfonate (136 mg, 0.234 mmol, Example 41), 3-dimethylamino-l-propyne (39 mg, 0.468 mmol, 2 equ), copper iodide (1 mg, 0.005 mmol, 0.002 equ) and triethylamine (0.082 mL, 0.585 mmol, 2.5 equ) were stirred in N,N-dimethylformamide (2 mL) under nitrogen in a microwave vial. Tetrakis(triphenylphosphine)palladium(0) (14 mg, 0.012 mmol, 0.05 equ) was added and nitrogen was bubbled through the mixture before being heated at 80°C under microwave radiation for 5 mins (300 W, 300 psi). The residue was adsorbed onto silica and purified by flash silica chromatography (gradient elution DCM:MeOH 100%-99%-98%-97%- 96%-95%-94%-93%-92%-91%-90%) to yield an orange solid (98 mg), which was then re- purified by preparative TLC (eluent 10:1 DCM: MeOH) to give the title compound as a yellow solid (52 mg, 43% yield).
<¾ (DMSO-de): 9.01 (1H, d, J= 1.6), 8.87-8.89 (1H, dd, J= 8.5, 0.6), 8.58-8.59 (1H, dd, J= 8, 1), 8.40-8.44 (1H, dd, J1=J2= 8.1), 8.37-8.39 (1H, d, J= 8), 8.08-8.12 (1H, ddd, J= 9.8, 7.1, 1.2), 8.02-8.07 (2H, ddd, 10.2, 9.5, 1.7), 7.92-7.95 (1H, dd, J- 8.5, 1.0), 7.70-7.74 (1H, ddd, J= 8.4, 7.3, 1.1), 4.62 (3H, s), 3.62 (2H, br s), 2.36 (6H, br s).
m/z (ES+): 365.1 (M +).
Example 45 - Preparation of 3-(3-acetamidoprop-l-vnyl)-13-methylchromeno[4,3,2- g¾lphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000116_0002
13-Methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/z]phenanthridin-13-ium
trifluoromethanesulfonate (136 mg, 0.234 mmol, Example 41), N-(prop-2-ynyl)acetamide (45 mg, 0.468 mmol, 2 equ), copper iodide (1 mg, 0.005 mmol, 0.002 equ) and triethylamine (0.082 mL, 0.585 mmol, 2.5 equ) were stirred in N,N-dimethylformamide (2 mL) under nitrogen in a microwave vial. Tetrakis(triphenylphosphine)palladium(0) (14 mg, 0.012 mmol, 0.05 equ) was added and nitrogen was bubbled through the mixture before being heated at 80°C under microwave radiation for 5 mins (300 W, 300 psi). The residue was adsorbed onto silica and purified by flash silica chromatography (gradient elution DCM:MeOH 100%-99%-98%-97%- 96%-95%-94%-93%-92 -91%-90%) to yield an orange solid (95 mg), which was then re- purified by preparative TLC (eluent 10:1 DCM: MeOH) to give the title compound as a yellow solid (67 mg, 54% yield).
4 (DMSO- ): 8.98 (1H, d, J= 1.8), 8.84-8.86 (1H, dd, J= 8.4, 0.7), 8.56-8.58 (1H, dd, J= 8.5, 1.0), 8.48-8.51 (1H, m), 8.36-8.43 (3H, m), 8.08-8.13 (1H, ddd, J= 8.5, 7.1, 1.3), 8.03-8.04 (1H, dd, J= 8.3, 0.6), 7.91-7.94 (1H, dd, J= 8.5, 1.1), 7.70-7.73 (1H, ddd, J= 8.1, 7.7, 0.9), 4.61 (3H, s), 4.23-4.24 (2H, d, J= 5.5), 1.90 (3H, s).
m/z (ES+): 379.2 (M +). Example 46 - Preparation of 3,ll-dimethoxy-13-methylchromenor4,3,2-gfelphenanthridin-
13-ium trifluoromethanesulfonate
Step 1 - Preparation of 3-(2-bromo-4-methoxyphenoxy)-2'-fluoro-5'-methoxybiphenyl-2- carbonitrile
Figure imgf000117_0001
2',3-Difluoro-5'-methoxybiphenyl-2-carbonitrile (2.50 g, 10.19 mmol, Example 40), 2-bromo-4- methoxyphenol (3.10 g, 15.29 mmol, 1.5 equ) and potassium carbonate (2.82 g, 20.38 mmol, 2 equ) were stirred in N,N-dimethylformamide (30 mL) at 100°C under nitrogen for 24 hours. The reaction was cooled to room temperature and a saturated solution of ammonium chloride (100 mL) was added. The mixture were extracted with diethyl ether (3 x 100 mL) and the combined organic extracts were washed with brine (50 mL), dried over magnesium sulfate and concentrated to yield a straw coloured oil, which was triturated with an hexane / diethyl ether mixture to yield a white solid. The solid was then triturated with heptane:EtOAc 4:1, filtered and air dried to give the title compound as a white solid (3.66 g, 84% yield).
<¾ (DMSO-d6): 7.66-7.70 (1H, ddd, d= 8.5, 7.7, 0.7), 7.40 (1H, s), 7.38-7.40 (1H, d, J= 5.6), 7.32-7.37 (1H, dd, J= 9.2), 7.27-7.29 (1H, dd, J= 8.5, 0.8), 7.06-7.13 (3H, m), 6.70-6.72 (1H, dd, J= 8.5, 0.7), 3.83 (3H, s), 3.81(3H, s). m/z (ES+): 428.0 (MH+, 79Br), 430.0 (MH+, 81Br).
Step 2 - Prepa
Figure imgf000118_0001
Tert-butyl lithium (1.6M in pentane, 11.21 mL, 17.95 mmol, 2.1 equ) was added to tetrahydrofuran (50 mL) at -78°C and the mixture was stirred for 5 mins. 3-(2-bromo-4- methoxyphenoxy)-2'-fluoro-5'-methoxybiphenyl-2-carbonitrile (3.66 g, 8.55 mmol, 1 equ) dissolved in THF (20 mL) was added drop wise. The resulting deep red solution was stirred at - 78°C for 5 min and the reaction was stirred at room temperature for 30 mins. Sat ammonium chloride (100 mL) was added and the mixture was extracted with dichloromethane (3 x 50 mL). The combined organic extracts were combined, dried over magnesium sulphate and concentrated in vacuo. The yellow solid was triturated with diethyl ether and the precipitate filtered from solution and air dried. This was then re-crystallised from hexane:EtOAc 4: 1 , to give the title compound as a pale yellow solid (2.2 g, 78% yield).
4 (DMSO- ): 8.42-8.44 (1H, d, J= 7.9), 8.04 (1H, d, J= 2.7), 7.97-8.0 (2H, m), 7.90-7.94 (1H, dd, J= 8.1), 7.47-7.48 (1H, dd, J= 7.8, 0.5), 7.37-7.41 (2H, m), 7.20-7.23 (1H, dd, J= 9.0, 3.1 , 4.00 (3H, s), 3.91 (3H, s).
m/z (ES+): 330.1 (M+H+).
Step 3 - Preparation of 3,ll-dimethoxy-13-methylchromeno[4,3,2- ¾]phenanthridin-13- ium trifluoromethanesulfonate
Figure imgf000118_0002
A mixture of 3,l l-dimethoxychromeno[4,3,2-g i]phenanthridine (100 mg, 0.304 mmol) potassium carbonate (85 mg, 0.608 mmol, 2 equ) and methyl trifluoromethanesulfonate (0.69 mL, 6.07 mmol, 20 equ) in chloroform (10 mL) was heated at 110°C in a sealed tube for 72 hours. The reaction was cooled to room temperature, quenched with methanol and was transferred to a flask where a saturated solution of NaHCC>3 was added slowly with stirring. The product was extracted with dichloromethane and the combined organic extracts were dried over magnesium sulphate, filtered and concentrated to give a yellow solid. The solid was adsorbed onto silica and purified by flash silica chromatography (gradient elution, hexane:EtOAc 97%- 94%-90%to DCM:MeOH 97%-95%) to give the title compound as an orange solid (106 mg, 71% yield).
4 (DMSO-de): 8.77-8.79 (1H, d, J- 7.8), 8.34-8.38 (1H, dd, J= 8.16), 8.28-8.32 (2H, dd, J= 9.4, 2.8), 7.95-7.97 (1H, dd, J= 8.4, 0.5), 7.84-7.87 (2H, m)7.68-7.71 (1H, dd, J= 9.2, 2.8), 7.64-7.67 (1H, dd, 9.4, 2.7), 4.66 (3H, s), 4.07 (3H, s), 3.98 (3H, s).
m/z (ES+): 314.1 (M+).
Example 47 - Preparation of 3,ll-difluoro-13-methyl-2-(lH-pyrazol-4-Yl)chromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000119_0001
A mixture of 3,ll-difluoro-2-methoxychromeno[4,3,2- z]phenanthridine (178 mg, 0.531 mmol, Example 52) and pyridine hydrochloride (~3g, ~ 25 mmol, ~ 50 equ) in 3 separate vials were heated under microwave radiation at 210°C (300W, 200psi, cooling system on) for 10 min. The combined reaction mixtures were poured into ice and the resulting precipitate was filtered and washed successively with water and heptane and then dried under suction to give 3,11- difluorochromeno[4,3,2-g¾]phenanthridin-2-ol as a red solid (148 mg, 0.461 mmol, 87% yield), (m/z (ES+): 322.0 (MH+). The material was used without further characterization and purification in the next steps.
To a solution of 3,ll-difluorochromeno[4,3,2-^¾]phenanthridin-2-ol (148 mg, 0.461 mmol) in 1:1 CHCl3:THF (2.5 mL) under a nitrogen atmosphere at room temperature was added N- Phenyl-bis(trifluoromethanesulfonimide) (494 mg, 1.382 mmol, 3.0 equ) followed by the addition of Ν,Ν-diisopropylethylamine (0.241 mL, 1.382 mmol, 3.0 equ). A condenser was fitted and the reaction mixture was placed in a pre-heated oil batch at 55°C. After 12 h, the reaction mixture was allowed to cool to room temperature and transferred to a separating funnel when hyrdrochloric acid (1M, 25 mL) was added and the crude solution was extracted with DCM (4 x 20 mL). The combined organic extracts were washed with saturated sodium bicarbonate solution (20 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. The crude material was then absorbed onto silica and subjected to flash chromatography (gradient elution Hexane:EtOAc 100%-98%-96%-92%-90%-85%-80%) to give 3,1 l-difluorochromeno [4,3,2- g/z]phenanthridin-2-yl trifluoromethanesulfonate as a slightly off-white solid (185 mg, 0.408 mmol, 89% yield), (m/z (ES+): 454.0 (MH+). The material was used without further characterization and purification in the next steps.
A mixture of 3,l l-difluorochromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate (185 mg, 0.408 mmol), methyl triflate (0.5 mL, 4.081 mmol, 10.0 equ) and potassium carbonate (570 mg, 4.081 mmol, 10.0 equ) in anhydrous chloroform (5 mL) was heated at 110°C in a sealed tube. After 24 h the reaction was cooled to room temperature and diluted with methanol and the insoluble material was removed by filtration. The filtrate was absorbed on silica and the product was purified by flash chromatography (gradient elution DCM:MeOH 100%-95%-90%) to give 3,l l-difluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-^]phenanthridin-13- ium trifluoromethanesulfonate (140 mg, 0.227 mmol, 56%) (m/z (ES+): 468.0 (MH+) as an orange solid. The material was used without further characterization and purification in the next steps.
A mixture of 13,l l-difluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2- g j]phenanthridin-13-ium trifluoromethanesulfonate (30 mg, 0.049 mmol), lH-pyrazole-4- boronic acid pinacol ester (14 mg, 0.073 mmol, 1.5 equ), tetrakis(triphenylphosphine)palladium(0) (2.8 mg, 0.0024 mmol, 0.05 equ) and sodium acetate (8.8 mg, 0.107 mmol, 2.2 equ) in 3:1 DME:H20 (5 mL) was heated at 90°C under MW radiation (30s. ramp time, 300W, 200psi, cooling system on) for 25min. 0.5equ of boronic ester, 0.05 equ of tetrakis(triphenylphosphine)palladium(0) and 2.2 equ of sodium acetate were added and the mixture was re-heated under MW radiation at 90°C (30s. ramp time, 300W, 200psi, cooling system on) for 20min. A reaction mixture of a separate experiment carried out on 5 mg scale was combined and the mixture was concentrated in vacuo. The resulting solid was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 100%-98%-97%-96%-94%-92%-90%) to give 8 mg of an orange solid. The solid was triturated with IN HC1, filtered, washed successively with water and diethyl ether, then dried under suction to give the title compound as an orange solid (5.7 mg, 0.011 mmol, 24% yield). <¾ (DMSO- e): 13.46 (1H, s), 8.91-8.94 (1H, d, J=12.2), 8.76-8.76 (1H, d, J=8.2), 8.58-8.60 (1H, m), 8.56-8.58 (1H, d, J=6.8), 8.40-8.44 (1H, t, 8.2), 8.36-8.38 (1H, m), 7.99-8.04 (3H, m), 7.67 (1H, s), 4.73 (3H, m).
m z (ES+): 386.0 (M+).
Example 48 - Preparation of 3-fluoro-2-(methoxycarbonyl)-13-methylchromenor4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of 2-bromo-4-fluoro-5-methoxy-N-(9H-xanthen-9-ylidene)aniline
Figure imgf000121_0001
A IM solution of titanium tetrachloride in toluene (47 mL, 47 mmol, 5 equ) was added dropwise to a suspension of 9H-xanthen-9-one (1.85 g, 9.0 mmol) in toluene (200 mL) and stirred at 25°C. After 0.5 h, 2-bromo-4-fluoro-5-methoxyaniline (2.5 g, 10.0 mmol, 1.2 equ) was added in one portion and the resulting orange suspension was heated to 110°C for 1 h when N,N diisopropylethylamine (8.2 mL, 47 mmol, 5 equ) was added. After a further 0.5 h, the reaction mixture was cooled when methanol (5 mL) was added and the crude material was absorbed on silica and purified by flash chromatography (gradient elution CHCl3:DCM, 100%-0%) to give the title compound as a yellow solid (600 mg, 1.5 mmol, 16% yield),
m/z (ES+): 397.0 (MH+, 79Br), 399.0 (MH+, 81Br).
Step 2 - Preparation of 3-fluorochromeno[4,3,2- A]phenanthridin-2-ol
Figure imgf000121_0002
A mixture of 2-bromo-4-fluoro-5-methoxy-N-(9H-xanthen-9-ylidene)aniline (560 mg, 1.4 mmol), sodium t-butoxide (400 mg, 4.1 mmol, 3 equ), 2-diphenylphosphino-2'-(N,N- dimethylamino)biphenyl (42 mg, 0.012 mmol, 0.08 equ), and bis(triphenylphosphine)palladium(II) dichloride (40 mg, 0.006 mmol, 0.04 equ) in anhydrous dimethylacetamide (10 mL) was heated at 180°C under microwave irradiation for 10 min. (300W, 300psi, with cooling system on). The residue was concentrated in vacuo as a mixture of 3-fluoro-2-methoxychromeno[4,3,2-g z]phenanthridine (m/z (ES+): 317.0 (MH+) contaminated with 9H-xanthen-9-one (m/z (ES+): 196.0 (MH+) impurity. The material was used without further characterization and purification in the next steps.
A mixture of crude 3-fluoro-2-methoxychromeno[4,3,2-g z]phenanthridine and pyridine hydrochloride (500 mg, 4.3 mmol, ~3 equ) was heated under microwave radiation at 215°C (200W, lOOpsi, cooling system on) for 5 min. The reaction mixture was poured into ice and the resulting precipitate was filtered and then absorbed on silica, and was purified by flash chromatography (gradient elution DCM:MeOH 98 -95%) to give the title compound as an off- white solid (270 mg, 0.89 mmol, 63% yield over 2 steps).
<¼ (CDCIJ): 10.72 (1H, s), 8.52-8.54 (1H, dd, J=7.9, 1.6), 8.42-8.45 (1H, d, J=12.6), 8.32 (1H, s), 8.25-8.27 (1H, d, J=8.0), 7.86-7.90 (1H, t, J=8.1), 7.62-7.97 (1H, ddd, J=8.6, 7.1, 1.7), 7.50- 7.53 (1H, d, J=8.8), 7.38-7.45 (3H, m).
(m z (ES+): 303.0 (MH+).
Step 3 - Preparation of 3-fluorochromeno[4,3,2-g/?]phenanthridin-2-yl
trifluoromethanesulfonate
Figure imgf000122_0001
To a solution of 3-fluorochromeno[4,3,2-g i]phenanthridin-2-ol (200 mg, 0.66 mmol) in 1:1 CHC iTHF (10 mL) under a nitrogen atmosphere at room temperature was added N-Phenyl- bis(trifluoromethanesulfonimide) (350 mg, 1.0 mmol, 1.5 equ) followed by the addition of Ν,Ν- diisopropylethylamine (0.175 mL, 1.0 mmol, 1.5 equ). The reaction mixture heated at 60°C and after 24 h, the reaction mixture was concentrated in vacuo. The crude material was then absorbed onto silica and subjected to flash chromatography (CHCI3) to give the title compound as a white solid (65 mg, 0.15 mmol, 23% yield).
<¾ (CDClj): 8.76 (1H, br s), 8.21-8.24 (1H, d, J=10.7), 8.12-8.20 (1H, m), 8.04-8.06 (1H, d, J=7.9), 7.91-7.96 (1H, t, J=8.1), 7.61-7.65 (1H, m), 7.50-7.54 (1H, d, 7.8), 7.38-7.43 (2H, m). (m z (ES+): 435.0 (MH+).
Step 4 - Preparation of 3-fluorochromeno[4,3,2-gA]phenanthridine-2-carboxylate
Figure imgf000123_0001
3-Fluorochromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate (125 mg, 0.29 mmol), palladium acetate (1.2 mg, 0.012 mmol, 0.02 equ) and Xantphos (6.5 mg, 0.011 mmol, 0.04 equ) were put in vacuo in a round bottom flask and the flask was filled with carbon monoxide. A 2:1 mixture of degassed N,N-dimethylformamide:methanol (3 mL) and triethylamine (0.080 mL, 0.57 mmol, 2.0 equ) was added and the mixture was heated to 60°C for 1.5 h. After cooling, the crude material was absorbed on silica and the product was purified by flash chromatography (DCM) to give the title compound as an off-white solid (80 mg, 0.23 mmol, 81% yield).
<¾ (DMSO-d6): 8.61-8.64 (1H, d, J=12.2), 8.55-8.58 (1H, dd, J=8.1, 1.7), 8.46-8.50 (2H, m), 8.00-8.04 (1H, t, J=8.2), 7.68-7.70 (1H, ddd, J=8.6, 7.2, 1.8), 7.64-7.70 (1H, d, J=8.0), 7.48-7.50 (1H, dd, J=7.6, 0.8), 7.42-7.46 (1H, m), 3.97 (3H, s)
m/z (ES+): 345.0 (MH+)
Step 5 - Preparation of 3-fluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2-
Figure imgf000123_0002
A mixture of methyl 3-fluorochromeno[4,3,2-g/i]phenanthridine-2-carboxylate (75 mg, 0.22 mmol), methyl triflate (0.119 mL, 1.10 mmol, 5.0 equ) and potassium carbonate (45 mg, 0.30 mmol, 1.5 equ) in anhydrous chloroform (10 mL) was heated at 115°C in a sealed tube. After 36 h the reaction was cooled to room temperature and diluted with methanol and the insoluble material was removed by filtration. The filtrate was absorbed on silica and the product was purified by flash chromatography (DCM:MeOH 90%) to give the title compound as an orange solid (110 mg, 0.22 mmol, 100% yield).
<¼ (DMSO-d6): 8.98-9.00 (1H, d, J=11.4), 8.82-8.84 (1H, d, J=7.7), 8.72-8.74 (1H, d, J=6.1), 8.56-8.59 (1H, dd, 8.9, 1.7), 8.46-8.51 (1H, t, J=8.2), 8.12-8.16 (2H, m), 7.95-7.98 (1H, dd, 8.5, 1.0), 7.74-7.78 (1H, ddd, J=8.3, 7.0, 1.1), 4.67 (3H, s), 4.03 (3H, s).
m/z (ES+): 360.0 (MH+) Example 49 - Preparation of 3,ll-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- g¾lphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of methyl 3 1-difluorochromeno[4,3,2-gft]phenanthridine-2- carboxylate
Figure imgf000124_0001
3,ll-Difluorochromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate (100 mg, 0.221 mmol, Example 47), palladium acetate (1 mg, 0.0044 mmol, 0.02 equ) and Xantphos (5 mg, 0.0088 mmol, 0.04 equ) were put in vacuo in a round bottom flask and the flask was filled with carbon monoxide. A 2:1 mixture of degassed N,N-dimethylformamide:methanol (5 mL) and triethylamine (0.092 mL, 0.663 mmol, 3.0 equ) was added and the mixture was stirred at 60°C. After 1 h, the reaction mixture was cooled and the resulting black solid was suspended in DCM:MeOH and the insoluble material was removed by filtration. The filtrate was absorbed on silica and purified by flash chromatography (gradient elution DCM:EtOAc 100%-80%) to give the title compound as an off-white solid (14 mg, 0.039 mmol, 18% yield).
4 (CDClj): 8.81 (1H, br s), 8.47 (1H, br s), 8.11 (1H, s), 8.08-8.09 (1H, d, J=3.0), 7.89-7.93 (1H, t, J=8.2), 7.48-7.52 (1H, m), 7.30-7.36 (2H, m), 4.03 (3H, s).
m/z (ES+): 364.0 (MH+)
Step 2 - Preparation of 3,ll-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000124_0002
A mixture of methyl 3,l l-difluorochromeno[4,3,2- /z]phenanthridine-2-carboxylate (28 mg, 0.076 mmol), methyl triflate (0.043 mL, 0.286 mmol, 5.0 equ) and potassium carbonate (12 mg, 0.076 mmol, 1.0 equ) in anhydrous chloroform (5 mL) was heated at 110°C in a sealed tube. After 24 h the reaction was cooled to room temperature and diluted with methanol and the insoluble material was removed by filtration. The filtrate was absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 100%-95%-90%) and the resulting solid was sonicated in ethyl acetate/ether to give the title compound as an orange solid (10 mg, 0.019 mmol, 56% yield).
& (CDjOD): 8.82-8.83 (1H, d, J=6.0), 8.71-8.75 (2H, m), 8.44-8.48 (1H, t, J=8.2), 8.34-8.37 (1H, dd, J=8.9, 1.8), 8.08-8.10 (1H, d, J=8.2), 7.96-8.01 (2H, m), 7.75 (3H, s), 4.07 (3H, s). m/z (ES+): 378.0 (MH+)
Example 50 - Preparation of 2-carbamoyl-l,ll-difluoro-13-methylchromenor4,3,2- gfelphenanthridin-13-ium chloride
Step 1 - Preparation of methyl l,ll-difluorochromeno[4,3,2-gft]phenanthridine-2-
Figure imgf000125_0001
l,ll-Difluorochromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate (330 mg, 0.728 mmol, Example 18), palladium acetate (8.2 mg, 0.037 mmol, 0.05 equ) and xantphos (42 mg, 0.0728 mmol, 0.1 equ) were put in vacuo in a round bottom flask and the flask was filled with carbon monoxide. A 2:1 mixture of degassed N,N-dimethylformamide:methanol (10 mL) and triethylamine (203 iL, 1.46 mmol, 2 equ) were added and the mixture was stirred at 60°C. After 1 h, a solution of palladium acetate (8.2 mg, 0.037 mmol, 0.05 equ) and xantphos (42 mg, 0.0728 mmol, 0.1 equ) in degassed Ν,Ν-dimethylformamide was added and the mixture was stirred at 60°C under carbon monoxide atmosphere. After 3 h, 0.025 equivalents of palladium acetate and 0.05 equivalents of xantphos dissolved in N,N-dimethylformamide were added and the mixture was stirred at 60°C for 16 h under carbon monoxide atmosphere. A mixture from a separate experiment carried out on 50 mg scale was combined and the mixture was concentrated to dryness. The resulting black solid was suspended in DCM:MeOH and the insoluble material was removed by filtration. The filtrate was absorbed on silica and the product was purified by flash chromatography (DCM) to give the title compound as an off-white solid (94 mg, 31% yield). <¾ (CDClj): 8.39-8.42 (1H, ddd, J=8.8, 2.8, 0.5), 8.15-8.19 (2H, m), 8.01-8.05 (1H, dd, J=8.6, 6.6), 7.87-7.91 (1H, dd, J1=J2=8.0), 7.28-7.34 (3H, m), 4.05 (3H, s).
m/z (ES+): 364.1 (MH+), 386.1 (M+Na+) Step 2 - Preparation of 2-carbamoyl-± 1-difluoro-13-methylchromeno[4,3,2- g/i]phenanthridin-13-ium chloride
Figure imgf000126_0001
A mixture of methyl l,l l-difluorochromeno[4,3,2-g z]phenanthridine-2-carboxylate (94 mg, 0.259 mmol), methyl triflate (142 μΐ^, 1.29 mmol, 5 equ) and potassium carbonate (54 mg, 0.389 mmol, 1.5 equ) in chloroform (5 mL) was heated at 115°C in a sealed tube for 72 h. The reaction was quenched with methanol and the insoluble material was removed by filtration. The filtrate was absorbed on silica and the product was purified by flash chromatography (gradient elution DCM:MeOH 100%-95%). The product was only partially purified and the mixed fractions were dissolved in DCM:MeOH, absorbed on silica and repurified by flash chromatography (gradient elution DCM:MeOH 100%-99%-98%-97%-95%) to give 87 mg of an orange solid and 29 mg of recovered starting material. The solid was triturated in 10% Et20:MeOH, filtered, washed with 1:1 Et20:EtOAc then diethyl ether and dried under suction to give l,ll-difluoro-2- (methoxycarbonyl)-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium
trifluoromethanesulfonate (36 mg) as an orange solid. NMR analysis of this product showed that it was -85% pure and was used without further purification and characterisation in the next step. A suspension of l,l l-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- g/i]phenanthridin-13-ium trifluoromethanesulfonate (60 mg, 0.114 mmol) in 3N HCI (10 mL) was heated at 85°C overnight. The mixture was concentrated to dryness in vacuo to give 2- carboxy-l,ll-difluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium chloride as a yellow solid, which was used without further purification in the next step. A suspension of crude 2- carboxy- 1 , 11 -difluoro- 13 -methylchromeno[4,3,2-g/i]phenanthridin- 13-ium chloride in phosphorus oxychloride (5 mL) was refluxed for 2 h and was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo and the product was co- evaporated with toluene and hexane to give 2-(chlorocarbonyl)-l,ll-difluoro-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium chloride as a brown/green solid and was used without purification in the next step.
Ammonia was condensed into a suspension of crude 2-(chlorocarbonyl)-l,ll-difluoro-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium chloride in anhydrous tetrahydrofuran (5 mL) at -78°C and the mixture was stirred for 20min. The mixture was allowed to warm to room temperature and was stirred for ~20min, then concentrated to dryness in vacuo. The resulting solid was suspended in DCM:MeOH. The insoluble material was removed by filtration and the filtrate was absorbed on silica, and the product was purified by flash chromatography (neutralized silica, DCM:Et3N 99%). The product was then co-evaporated successively with diethyl ether and ethyl acetate to give 15 mg of a yellow solid. The product was triturated with 1M HC1 and the resulting precipitate was filtered, washed with diethyl ether and dried under suction to give the product as an orange/yellow solid (5 mg).
The filtrate was concentrated in vacuo and tetrahydrofuran followed by a small amount of diethyl ether were added. The resulting suspension was left for 72 h before being filtered. The solid was washed with diethyl ether and dried under suction to give the title compound as a yellow/orange solid (6 mg, 24% yield).
4 (DMSO-d6): 8.80-8.82 (1H, d, J=8.0), 8.74-8.77 (1H, d, J=8.6), 8.62-8.65 (1H, dd, J=10.0, 2.3), 8.49-08.53 (1H, d, J=8.4), 8.21 (1H, s (br)), 8.05-8.15 (4H, m), 8.00-8.04 (1H, dd, J=8.4, 6.3), 4.62-4.64 (3H, d, J=9.6).
m/z (ES+): 363.0 (M+)
Example 51 - Preparation of 2-carboxy-13-methylchromenor4,3,2-g¾lphenanthridin-13- ium trifluoromethanesulfonate
Figure imgf000127_0001
A mixture of 2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (700 mg, 1.4 mmol; Example 20) in 30% aqueous TFA (15 mL) was heated at 90°C. After 16 h, the mixture was concentrated to dryness in vacuo and the product was purified by flash chromatography (DCM:MeOH 85%) to give the title compound as a yellow solid (500 mg, 74% yield).
<¾ (DMSO-d6): 8.94-8.96 (1H, d, J=8.6), 8.77-8.79 (1H, d, J=8.3), 8.74 (1H, d, J=1.2), 8.57-8.60 (1H, dd, J=8.4, 1.2), 8.42-8.46 (1H, dd, J1=J2=8.1), 8.34-8.37 (1H, dd, J8.4), 8.09-8.13 (1H, ddd, J=8.5, 7.2, 1.3), 8.04-8.06 (1H, d, J=8.4), 7.91-7.94 (1H, dd, J=8.5, 1.3), 7.71-7.75 (1H, ddd, J=8.3, 7.2, 1.3), 4.69 (3H, s).
m/z (ES+): 328.0 (M+). Example 52 - Preparation of 3,ll-difluoro-2-methoxy-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
- Preparation of 2',3,5'-trifluoro-4'-methoxybiphenyl-2-carbonitrile
Figure imgf000128_0001
A suspension of 4-bromo-2,5-difluoroanisole (4.91 g, 22.0 mmol), potassium acetate (6.49 g, 66.0 mmol, 3 equ) and bis(pinacolato)diboron (6.15 g, 24.2 mmol, 1.1 equ) in anhydrous N,N- dimethylformamide (100 mL) was degassed for 5 mins before the addition of 1,1'- bis(diphenylphosphino)ferrocene-palladium(II)dichloride acetone complex (365 mg, 0.462 mmol, 0.02 equ) and the reaction mixture was heated at 100°C for 45 mins. 0.02 equ of 1,1'- bis(diphenylphosphino)ferrocene-palladium(II)dichloride acetone complex followed by a 2M sodium carbonate solution (20 mL) and 2-bromo-6-fluorobenzonitrile (5.28 g, 26.4 mmol, 1.2 equ) were added and the reaction mixture was stirred at 100°C under nitrogen for 1 nr. The reaction mixture was allowed to cool to room temperature and was concentrated to dryness in vacuo. The residue was dissolved in dichloromethane and filtered through celite. The filtrate was absorbed on silica and the product was purified by flash chromatography (hexane:EtOAc 20%) to give the title compound as a white solid (1.14 g, 20% yield over 2 steps).
<¾ (CDClj): 7.63-7.68 (1H, m), 7.24-7.29 (3H, m), 7.14-7.19 (1H, dd, J=10.9, 6.9).
m/z (ES+): 264.0 (MH+), 286.0 (M+Na+)
Step 2 - Preparation of 3-(2-bromo-4-fluorophenoxy)-2',5'-difluoro-4'-methoxybiphenyl-2- carbonitrile
Figure imgf000128_0002
A suspension of 2',3,5'-trifluoro-4'-methoxybiphenyl-2-carbonitrile (1.06 g, 4.03 mmol), 2- bromo-4-fluorophenol (1.16 g, 6.04 mmol, 1.5 equ) and potassium carbonate (1.12 g, 8.05 mmol, 2 equ) in anhydrous N,N-dimethylformamide (15 mL) was heated under nitrogen at 140 °C. After 45 min., 1.5equ of 2-bromo-4-fluorophenol were added and the mixture was heated at 140°C for 45min. The cooled reaction mixture was poured into a saturated ammonium chloride solution (~70mL) and the mixture was transferred into a separating funnel. The product was extracted with diethyl ether (5 x 40 mL) and the combined organic extracts were washed with brine (40 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. The pale brown residue was dissolved in dichloromethane, absorbed onto silica and purified by flash chromatography (hexane:EtOAc 20%) to give the title compound as a white solid (1.36 g, 78% yield).
4 (CDClj): 7.47-7.51 (1H, dd, J=8.5, 7.8), 7.43-7.46 (1H, dd, J=7.7, 2.9), 7.17-7.22 (2H, m), 7.10-7.15 (2H, m), 6.85-6.89 (1H, dd, J=ll.l, 7.0), 6.63-6.65 (1H, dd, J=8.5, 0.9), 3.96 (3H, s). Step 31 - Preparation of 3,ll-difluoro-2-methoxy-13-methylchromeno[4,3,2- g¾]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000129_0001
A solution of 3-(2-bromo-4-fluorophenoxy)-2',5'-difluoro-4'-methoxybiphenyl-2-carbonitrile (1.5 g, 3.45 mmol) in anhydrous tetrahydrofuran (6 mL) was added dropwise over 2 min. at - 78°C to a stirring solution of tert-butyl lithium (1.6M in pentane, 5.4 mL, 8.64 mmol, 2.5 equ) in anhydrous tetrahydrofuran (24 mL) under nitrogen, cooled to -78°C. After completion of the addition, the reaction mixture was allowed to warm to room temperature and was stirred overnight. The reaction was quenched with a saturated ammonium chloride solution (75 mL) and a reaction mixture from a separate experiment carried out on 1.9 g scale was combined. The mixture was transferred into a separating funnel and the product was extracted with dichloromethane (4 x 100 mL). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo to give 3,l l-difluoro-2-methoxychromeno[4,3,2- g j]phenanthridine as a pale yellow solid. The product (1 g) was used without further characterisation and purification in the next steps.
A suspension of 3,l l-difluoro-2-methoxychromeno[4,3,2-g z]phenanthridine (60 mg, 0.179 mmol), potassium carbonate (25 mg, 0.179 mmol, 1 equ) and methyl triflate (147 mg, 0.895 mmol, 5 equ) in chloroform (3 mL) was heated at 110°C in a sealed tube. After 16 h, chloroform (10 mL) was added and the insoluble inorganic material was removed by filtration. The filtrate was evaporated to dryness and the solid was triturated with 20% MeOH:DCM. The resulting suspension was filtered and the filtrate was evaporated to dryness in vacuo. The solid was dissolved in the minimum volume of 4% MeOH:DCM and the product was purified by flash chromatography (gradient elution DCM:MeOH 96%-88 ) to give the title compound as a bright orange solid (35 mg, 39% yield).
<¼ (DMSO-dg): 8.88-8.91 (IH, d, J=12.1), 8.64-8.67 (IH, d, J=8.4), 8.36-8.41 (IH, dd, J1=J2=8.0), 8.32-8.36 (IH, dd, J=9.5, 2.4), 7.96-8.05 (2H, m), 7.92-7.94 (IH, dd, J=8.2, 0.6), 7.85-7.67 (IH, d, J=7.6), 4.69 (3H, s), 4.20 (3H, s).
Example 53 - Preparation of 2-(ethoxycarbonvI)-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000130_0001
A mixture of 2-carboxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (130 mg, 0.27 mmol; Example 51) in phosphorus oxychloride (5 mL) was heated at 100°C for 1 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-13- methylchromeno[4,3,2-g¾phenanthridin-13-ium trifluoromethanesulfonate, which was used without further purification in the next step (quantitative yield assumed). Crude 2- (chlorocarbonyl)-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium trifluoromethanesulfonate was dissolved in pyridine (2 mL) and ethanol (2 mL) was added. The mixture was stirred at room temperature for 2 h and was concentrated to dryness in vacuo. The product was purified by flash chromatography (DCM:MeOH 90%) to give the title compound as a yellow/orange solid (120 mg, 87% yield).
<¾ (DMSO-d6): 9.04-9.06 (IH, d, J=8.9), 8.82-8.84 (IH, dd, J=8.3, 0.7), 8.75 (IH, d, J=1.3), 8.57-8.59 (IH, dd, J=8.7, 1.4), 8.46-8.50 (IH, dd, J1=J2=8.2), 8.36-8.39 (IH, dd, J=8.4, 1.4), 8.11-8.16 (IH, ddd, J=8.5, 7.2, 1.4), 8.11-8.13 (IH, dd, J=8.3, 0.6), 7.94-7.97 (IH, dd, J=8.5, 1.0), 7.73-7.77 (IH, ddd, J=8.3, 7.1, 1.4), 4.70 (3H, s), 4.46 (2H, q, J=7.1), 1.41-1.45 (3H, t, J=7.1).
Figure imgf000130_0002
m/z (ES+): 356.0 (M+). Example 54 - Preparation of 2-(isopropoxycarbonyl)-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000131_0001
A mixture of 2-carboxy-13-methylchromeno[4,3,2-gA]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.21 mmol; Example 51) in phosphorus oxychloride (2 mL) was heated at 100°C for 3 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate, which was used without further purification in the next step (quantitative yield assumed). Crude 2- (chlorocarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium trifluoromethanesulfonate was dissolved in pyridine (1 mL) and isopropanol (1 mL) was added. The mixture was stirred at room temperature for 16 h and was concentrated to dryness in vacuo. The product was subjected to purification by flash chromatography (DCM:MeOH 90%) to give the title compound as a yellow/orange solid (65 mg, 60% yield).
<¼ (DMSO-d6): 9.03-9.05 (IH, d, J=8.9), 8.82-8.84 (IH, d, J=8.3), 8.73 (IH, d, J=1.3), 8.58-8.60 (IH, dd, J=8.4, 1.1), 8.46-8.50 (IH, dd, J1=J2=8.1), 8.34-8.37 (IH, dd, J=8.5, 1.4), 8.11-8.15 (IH, ddd, J=8.5, 7.4, 1.4), 8.10-8.13 (IH, dd, J=8.3, 0.6), 7.94-7.97 (IH, dd, J=8.6, 1.1), 7.72- 7.77 (IH, ddd, J=8.3, 7.2, 1.1), 5.26-5.32 (IH, sep, 6.3), 4.70 (3H, s), 1.42-1.44 (6H, t, J=6.3).
Figure imgf000131_0002
m/z (ES+): 370.0 (M+).
Example 55 - Preparation of 2-carbamoyl-13-methylchromeno[4,3.2-gfelphenanthridin-13- ium trifluoromethanesulfonate
Figure imgf000131_0003
A mixture of 2-carboxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (13 mg, 0.27 mmol; Example 51) in phosphorus oxychloride (5 mL) was heated at 100°C for 1 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-13- methylchromeno[4,3,2- z]phenanthridin-13-ium trifluoromethanesulfonate, which was used without further purification in the next step (quantitative yield assumed). Crude 2- (chlorocarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate was dissolved in pyridine (5 mL) and the resulting solution was added to liquid ammonia at 0°C. The mixture was stirred at 0°C for 1 h and was then concentrated to dryness in vacuo. The product was purified by flash chromatography (DCM:MeOH 85%) to give the title compound as a mustard coloured solid (5 mg, 4% yield).
4 (DMSO-d6): 9.00-9.03 (1H, d, J=8.9), 8.81-8.83 (1H, d, J=8.3), 8.73-8.74 (1H, d, J=1.3), 8.58-8.60 (1H, dd, J=8.5, 1.2), 8.56 (1H, s, b(br)), 8.43-8.48 (1H, dd, J1=J2=8.2), 8.34-8.37 (1H, dd, J=8.6, 1.4), 8.09-8.14 (1H, ddd, J=8.6, 7.2, 1.4), 8.06-8.08 (1H, dd, J=7.1, 0.6), 7.93-7.95 (1H, dd, J=8.6, 1.1), 7.92 (1H, s (br)), 7.71-7.75 (1H, ddd, J=8.4, 7.0, 1.2), 4.68 (3H, s).
m/z (ES+): 327.1 (M+).
Example 56 - Preparation of 2-carbamovI-3-fluoro-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000132_0001
A suspension of 3-fluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g i]phenanthridin-13- ium trifluoromethanesulfonate (90 mg, 0.18 mmol; Example 48) in trifluoroacetic acid (1 mL) was heated at 100°C overnight. The mixture was concentrated in vacuo to give 2-carboxy-3- fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate as a yellow solid. The material was used without further characterization and purification in the next steps.
A suspension of crude 2-carboxy-3-fluoro-13-methylchromeno[4,3,2-g/?]phenanthridin-13-ium trifluoromethanesulfonate in phosphorus oxychloride (2 mL) was refluxed for 2 h and was allowed to cool to room temperature. The mixture was concentrated in vacuo and co-evaporated with toluene and hexane to give 2-(chlorocarbonyl)-3-fluoro-13-methylchromeno[4,3,2- gA]phenanthridin-13-ium chloride as a brown/green solid. The material was used without further characterization and purification in the next steps. Ammonia was condensed into a suspension of crude 22-(chlorocarbonyl)-3-fluoro-13- methylchromeno[4,3,2- z]phenanthridin-13-ium chloride in anhydrous chloroform (5 mL) and anhydrous pyridine (1 mL) at 0°C. After 2 h, the reaction mixture was allowed to warm to room temperature and then concentrated in vacuo, and the resulting solid was absorbed on silica, and purified by flash chromatography (DCM:MeOH 20%) to give the title compound as a yellow solid (30 mg, 0.061 mmol, 33% yield).
<¾ (DMSO-d6): 8.91-8.94 (1H, d, J=11.0), 8.80-8.82 (1H, d, J=7.8), 8.56-8.58 (2H, m), 8.44-8.48 (1H, t, J=8.2), 824 (1H, br s), 8.09-8.14 (3H, m), 7.94-7.96 (1H, dd, J=8.5, 1.0), 7.72-7.76 (1H, ddd, J=8.4, 7.3, 1.2), 4.67 (3H, s).
m/z (ES+): 346.0 (M+).
Example 57 - Preparation of 4,ll-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3^2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of 2',3,6'-trifluoro-4'-methoxybiphenyl-2-carbonitrile
Figure imgf000133_0001
A mixture of 2-bromo-6-fluorobenzonitrile (400 mg, 2.0 mmol), 2,6-difluoro-4- methoxyphenylboronic acid (452 mg, 2.4 mmol, 1.2 equ), bis(tri-tert- butylphosphine)palladium(O) (20 mg, 0.04 mmol, 0.02 equ) and potassium fluoride (382 mg, 6.6 mmol, 3.3 equ) in anhydrous tetrahydrofuran (4 mL) was heated under microwave radiation (300W, 150psi, cooling system on) at 120°C for 5 minutes. 8 consecutive runs were carried out and the reaction mixtures were combined, diluted with dichloromethane and transferred into a separating funnel. Water was added and the product was extracted with dichloromethane. The combined organic extracts were concentrated in vacuo and the resulting oil was absorbed on silica. The product was purified by flash chromatography (gradient elution hexane:EtOAc 99%- 95%) to give the title compound as a white solid (4.1 g, 81% yield).
<¾ (CDC13): 7.62-7.67 (1H, m), 7.23-7.27 (2H, m), 6.58-6.63 (2H, m), 3.87 (3H, s).
Step 2 - Preparation of 3-(2-bronio-4-fluorophenoxy)-2',6'-difluoro-4'-niethoxybiphenyl-2- carbonitrile
Figure imgf000134_0001
A mixture of 2',3,6'-trifluoro-4'-methoxybiphenyl-2-carbonitrile (1.11 g, 4.22 mmol, Example 57), 2-bromo-4-fluorophenol (1.21 g, 6.33 mmol, 1.5 equ) and potassium carbonate (1.16 g, 8.44 mmol, 2 equ) was stirred at 100°C for 8 h and was then allowed to cool to room temperature overnight. An additional 300 mg of 2',3,6'-trifluoro-4'-methoxybiphenyl-2-carbonitrile and 300 mg of potassium carbonate were added and the mixture was stirred at 100°C overnight. The mixture was cooled to room temperature and was poured into a saturated solution of ammonium chloride. The mixture was transferred into a separating funnel and the product was extracted with diethyl ether (4 x 100 mL). The combined organic extracts were washed with brine, dried over magnesium sulphate and concentrated in vacuo. The product was purified by flash chromatography (Hexane:EtOAc 25%) to give the title compound as a white solid (1.25 g, 68% yield).
<¾ (DMSO-i¾): 7.83-7.86 (1H, dd, J=8.0, 2.8), 7.69-7.73 (1H, dd, J=8.6, 7.7), 7.53-7.57 (1H, dd, J=9.0, 5.1), 7.40-7.45 (1H, ddd, J=8.9, 7.9, 3.0), 7.30-7.32 (1H, dd, J=7.6, 0.4), 6.97-6.99 (2H, d, J=9.1), 6.80-6.83 (1H, dd, J=8.6, 0.8), 3.87 (3H, s).
Step 3 - Preparation of 4,ll-difluoro-2-methoxychromeno[4,3,2-^ft]phenanthridine
Figure imgf000134_0002
fert-Butyl lithium (1.6M in pentane, 8.77 mmol, 2 equ) was added to anhydrous tetrahydrofuran (10 mL) at -78°C under nitrogen and the mixture was stirred for 5 mins before the slow addition of 3-(2-bromo-4-fluorophenoxy)-2',6'-difluoro-4'-methoxybiphenyl-2-carbonitrile (2.9 g, 6.68 mmol) in anhydrous THF (24 mL). The resulted red solution was stirred at -78°C for 5 minutes and was then allowed to room temperature overnight. The reaction mixture was cooled to 0°C then quenched by the addition of acetic acid (10 mL). The reaction mixture was stirred for 5min. and the pH of the solution was adjusted to 7-7.5 by the addition of IN sodium hydroxide. The mixture was transferred into a separating funnel and the product was extracted with dichloromethane. The combined organic extracts were washed with brine, dried over magnesium sulphate and concentrated in vacuo. The resulting orange solid was absorbed on silica and purified by flash chromatography (gradient elution Hexane:EtOAc 99%-98%) to give the pure title compound as a pale yellow solid (300 mg, 17% yield).
Notel: 1.4 g of partially purified title compound were obtained from mixed fractions, which could be used without further purification in the next step.
Note2: The pure product could also be obtained by precipitation in dichloromethane after extraction of the quenched reaction mixture.
<¾ (DMSO- 6): 8.37-8.39 (1H, d, J=8.3), 8.19-8.22 (1H, m), 7.95-7.99 (1H, ddd, J1=J2=8.2, J3=1.0), 7.55-7.58 (2H, m), 7.51-7.53 (1H, dd, J=8.1, 0.7), 7.40-7.41 (1H, dd, J=2.7, 0.6), 7.17- 7.22 (1H, dd, J=14.7, 2.6), 3.97 (3H, s).
m/z (ES+): 336.0 (MH+).
Step 4 - Preparation of 4,ll-difluorochromeno[4,3,2-g&]phenanthridin-2-yl
trifluoromethanesulfonate
Figure imgf000135_0001
A mixture of 4,ll-difluoro-2-methoxychromeno[4,3,2-g z]phenanthridine (170 mg, 0.507 mmol) and pyridine hydrochloride (-3 g, -26 mmol, -50 equ) was heated under microwave radiation at 210°C (300W, 200psi, cooling system on) for 5 min. 8 consecutive runs were carried out and the combined reaction mixture were poured into ice. The resulting precipitate was filtered, washed successively with water and hexane and was dried under suction to give 4,ll-difluoro-2- methoxychromeno[4,3,2-g/z]phenanthridine hydrochloride as a dark red/black solid (1.2g). The product was used without further purification and characterization in the next step.
N-Phenyl-bis(trifluoromethanesulfonimide) (1.6 g, 1.3 equ) followed by Ν,Ν- diisopropylethylamine (2.9 mL, 16.8 mmol, 5 equ) were added to a suspension of crude 4,11- difmoro-2-methoxychromeno[4,3,2-g/i]phenanthridine hydrochloride (1.2 g, 3.36 mmol) in a 1:1 DCM:THF mixture (200 mL) and the mixture was stirred at 60°C for 8h., then at room temperature. After 16 h, an additional 1.3 equ of N-Phenyl-bis(trifluoromethanesulfonimide) followed by 5 equ of Ν,Ν-diisopropylethylamine were added and the mixture was stirred at 60°C for 8 h., then at room temperature. After a further 16 h, the reaction mixture was concentrated to dryness in vacuo. The residue was absorbed on silica and the product was purified by flash chromatography (gradient elution Hexane:DCM 95%-90%-75%) to give the title compounds as a pale yellow solid (225 mg, 12% yield over 2 steps).
4 (DMSO-i¾): 8.44-8.47 (1H, d, J=8.3), 8.14-8.17 (1H, dd, J=8.8, 2.9), 8.05-8.09 (1H, dd, J=8.0), 7.94 (1H, m), 7.88-7.91 (1H, dd, J=12.6, 2.7), 7.66-7.68 (1H, d, J=8.1), 7.56-7.64 (2H, m).
Step 5 - Preparation of 4,ll-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000136_0001
Palladium acetate (2.2 mg, 0.0099 mmol, 0.02 equ), followed by 4,5-bis(diphenylphosphino)- 9,9-dimethylxanthene (12 mg, 0.0199 mmol, 0.04 equ) were added to a suspension of 4,11- difluorochromeno[4,3,2-g/z]phenanthridin-2-yl trifluoromethanesulfonate (225 mg, 0.490 mmol) and triethylamine (151 mg, 1.49 mmol, 3 equ) in degassed 2:1 N,N- dimethylformamide: methanol (20 mL). The mixture was evacuated with carbon monoxide. The mixture was stirred at room temperature for 30min. then at 60°C for 1 h under carbon monoxide atmosphere. An additional 0.02 equ of palladium acetate, 0.04 equ of 4,5- bis(diphenylphosphino)-9,9-dimethylxanthene and 3 equ of triethylamine were added and the mixture was stirred at 60°C for 1 h at room temperature, then at 60°C for 30 min. under a carbon monoxide atmosphere. The reaction mixture was allowed to cool to room temperature and was concentrated in vacuo. The resulting green residue was absorbed on silica and purified by flash chromatography (gradient elution Hexane:DCM 67%-50%-33%) to give methyl 4,11- difluorochromeno[4,3,2-g/z]phenanthridine-2-carboxylate as a pale yellow solid (50 mg, 28% yield) (m/z (ES+): 364 (MH+), 386 (M+ Na+). The material was used without further characterization or purification in the next steps. Note: Partially purified 4,l l-difluorochromeno[4,3,2-g z]phenanthridine-2-carboxylic acid (70 mg) could be obtained by washing the column with methanol.
A suspension of methyl 4,l l-difluorochromeno[4,3,2-g z]phenanthridine-2-carboxylate (50 mg, 0.137 mmol), methyl triflate (150 μί, 1.37 mmol, 10 equ) and potassium carbonate (57 mg, 0.413 mmol, 3 equ) in chloroform (1 mL) was heated at 115°C for 72 h and was allowed to cool to room temperature. The mixture was stirred at room temperature for 16 h, and then diluted with chloroform. The resulting suspension was filtered and the solid was triturated with a 10:1 DCM:MeOH mixture. The precipitate was removed by filtration and both filtrates were combined and were concentrated to dryness in vacuo. The product was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-90%). The product, which was obtained as a 2:1 mixture with an impurity identified as methyl 4,l l-difluoro-12b-hydroxy-13-methyl-12b,13-dihydrochromeno[4,3,2- g j]phenanthridine-2-carboxylate by LCMS analysis, was dissolved in DCM:MeOH, absorbed on silica and re -purified by flash chromatography (gradient elution DCM:MeOH 99%-98%-95%- 90%). The resulting solid was triturated in ethyl acetate, filtered and dried under suction to give the title compound as a yellow solid (9 mg, 12% yield).
<¼ (CDjOD): 8.93-8.96 (1H, ddd, J=8.3, 1.6, 0.6), 8.72 (1H, m), 8.48-8.52 (1H, ddd, J1=J2=8.2), J3=0.8), 8.35-8.38 (1H, dd, J=9.6, 2.5), 8.24-8.27 (1H, dd, J=12.7, 1.4), 8.13-8.15 (1H, dd, J=8.4, 0.7), 7.96-8.07 (2H, m), 4.74 (3H, s), 4.08 (3H, s).
m/z (ES+): 378.0 (M+)
Example 58 - Preparation of 2-carbamovI-3,ll-difluoro-13-methylchromeno[4,3,2- g¾lphenanthridin-13-ium chloride
Figure imgf000137_0001
A solution of 3,l l-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- z]phenanthridin- 13-ium trifluoromethane sulfonate (42 mg, 0.053 mmol; Example 49) in 3N hydrochloric acid (4 mL) was heated at 80°C. After 6 h, 1 mL of concentrated hydrochloric acid was added and stirred at 80°C. After a further 12 h, the reaction mixture was evaporated in vacuo to give 2- carboxy-3,l l-difluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium chloride (36 mg, 0.070 mmol, 88% yield) (m/z (ES+): 364.0 (MH ). The material was used without further characterization and purification in the next steps.
A suspension of 2-carboxy-3,ll-difluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium chloride (36 mg, 0.070 mmol) in phosphorous oxychloride (3 mL) was heated to 100°C for 2 h then allowed to cool to room temperature. The mixture was concentrated in vacuo and the product was co-evaporated with toluene and hexane to give 2-(chlorocarbonyl)-3,ll-difluoro-13- methylchromeno[4,3,2-g/ ]phenanthridin-13-ium chloride as a brown/green solid. The material was used without further characterization and purification in the next steps.
Ammonia was condensed into a suspension of crude 2-(chlorocarbonyl)-3,ll-difluoro-13- methylchromeno[4,3,2-g/ ]phenanthridin-13-ium chloride in anhydrous chloroform (6 mL) and anhydrous pyridine (0.2 mL) at room temperature. After 2 h, the reaction mixture was concentrated in vacuo, and the resulting solid was suspended in DCM:MeOH and the insoluble material was removed by filtration. The filtrate was absorbed on silica, and was purified by flash chromatography (gradient elution DCM:MeOH 90%-70%). The resulting solid was then purified by C18 reverse -phase chromatography (gradient elution water: methanol 50%-100%) to give the title compound as a red solid (6 mg, 0.015 mmol, 17% yield).
<¾ (CDjOD): 8.70-8.76 (3H, m), 8.43-8.47 (1H, t, J=8.2), 8.34-8.39 (1H, dd, J=9.4, 2.3), 8.06- 8.08 (1H, dd, J=8.3, 0.6), 7.93-8.01 (2H, m), 4.74 (3H, s).
m/z (ES+): 363.0 (M+).
Example 59 - Preparation of 13-methyl-2-(morpholinomethyl)chromenor4,3^2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000138_0001
Morpholine (103 mg, 1.1 mmol, 5.0 equ) was added to a suspension of 2-(bromomethyl)-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate (125 mg, 0.23 mmol, Example 26) in tetrahydrofuran (3 mL) at room temperature. The reaction mixture was stirred for 2 h at room temperature and sodium carbonate (100 mg) was added. The resulting suspension was stirred for 1 h and the inorganic materials were removed by filtration and washed with 20% MeOH:DCM. The filtrate was purified by flash chromatography (DCM:MeOH 90%) to give the title compound as a yellow solid (60 mg, 47% yield).
4 (CDjOD): 8.79-8.81 (1H, d, J=8.4), 8.65-8.68 (1H, d, J=8.6), 8.57-8.59 (1H, dd, J=8.5, 1.4), 8.36-8.40 (1H, dd, J1=J2=8.2), 8.34 (1H, d, J=0.9), 8.06-8.10 (1H, ddd, J=8.6, 7.1, 1.4), 7.93- 7.96 (2H, m), 7.87-7.90 (1H, dd, J=8.6, 1.2), 7.70-7.75 (1H, ddd, J=8.4, 7.1, 1.3), 4.73 (3H, s), 3.78 (2H, s), 3.75-3.77 (4H, m), 2.59-2.61 (4H, m).
Figure imgf000139_0001
Example 60 - Preparation of l,ll-difluoro-2-(2-hvdroxyethoxy)-13-methYlchromeno[4,3.i2- g/¾1phenanthridin- -ium methanesulfonate
Figure imgf000139_0002
A mixture of l,ll-difluoro-2-hydroxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium bromide (29 mg, 0.070 mmol; Example 63) and potassium carbonate (97 mg, 0.70 mmol, 10 equ) in N,N-dimethylformamide (2 mL) was stirred at room temperature for 10 min. before the addition of 2-iodoethanol (54 \L, 0.70 mmol, 10 equ). The mixture was stirred at 60°C for 1 h and was allowed to cool to room temperature. The reaction was quenched with acetic acid and the reaction mixtures from two separate experiments carried out on 10 mg scale were combined. The mixture was concentrated to dryness in vacuo and the residue was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-96%-94%-90%) to give 20 mg of a brown solid. The material was dissolved in 10% DCM:MeOH and Amberlyst resin, mesylate form (200 mg) was added. The mixture was stirred for 16 h and filtered. The filtrate was concentrated in vacuo and the resulting orange/brown solid was triturated with a 90% Et20:MeOH mixture. The precipitate was filtered, washed successively with 90% Ei20:MeOH, diethyl ether and dried under suction to give the title compound as an amber solid (8 mg, 14% yield).
<¼ (DMSO-i¾): 8.67-8.71 (2H, m), 8.58-8.61 (1H, dd, J=9.6, 2.3), 8.40-8.45 (1H, dd, J1=J2=8.3), 8.03-8.07 (2H, m), 7.97-7.99 (1H, d, J=8.1), 7.84-7.87 (1H, ddd, J=9.3, 8.1, 0.6), 5.04-5.07 (1H, t, J=5.3), 4.60-4.62 (3H, d, J=9.6), 4.37-4.39 (2H, t, J=4.6), 3.84-3.85 (2H, m). Example 61 - Preparation of 4-((13-methyIchromeno[4,3,2-jgMphenanthridin-13-ium-2- yl)methyl)morpholine -oxide trifluoromethanesulfonate
Figure imgf000140_0001
A solution of 0.05-0.1M DMDO in acetone (3 mL) was added to a solution of 13-mefhyl-2- (morpholinomethyl)chromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (15 mg, 0.02 mmol; Example 59) in acetone (2 mL) at room temperature and the mixture was stirred for 30 min., then concentrated in vacuo. The resulting solid was triturated with diethyl ether, filtered and dried under suction to give the title compound as a yellow solid (12 mg, 78% yield).
<¾ (DMSO-i¾: 8.91-8.94 (1H, d, J=8.5), 8.77-8.79 (1H, d, J=7.8), 8.62-8.64 (2H, m), 8.43-8.45 (1H, dd, J1=J2=8.2), 8.08-8.14 (2H, m), 8.02-8.04 (1H, d, J=8.2), 7.92-7.94 (1H, dd, J=8.5, 1.1), 7.69-7.74 (1H, ddd, J=8.5, 7.3, 1.1), 4.70-4.72 (2H, s (br)), 4.63 (3H, s), 4.09-4.15 (2H, m), 3.69- 3.73 (2H, m), 3.59-3.64 (2H, m), 2.76-2.79 (2H, m).
Example 62 - Preparation of 2-(2-amino-2-oxoethoxy)-13-methylchromeno[4,3.l2- gfelphenanthridin-13-ium formate
Figure imgf000140_0002
To a solution of 2-hydroxy-13-methylchromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (20 mg, 0.045 mmol; Example 6) in dimethylformamide (2 mL) was added potassium carbonate (61 mg, 0.445 mmol, 10 equ) and iodoacetamide (45 mg, 0.445 mmol, 10 equ) and the reaction was heated to 50°C. After 15 mins further amounts of 2- hydroxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (40 mg, 0.089 mmol, 2 equ), potassium carbonate (122 mg, 0.89 mmol, 20 equ), iodoacetamide (90 mg, 0.89 mmol, 20 equ) and dimethylformamide (1 mL) were added and the RM stirred for a further 0.5 h when the reaction mixture was allowed to cool then taken up in MeOH:DCM (10%), filtered with MeOH washings. To the filtrate was added glacial AcOH (0.5mL), which was then absorbed onto silica and purified by flash chromatography (gradient elution DCM:MeOH 100%- 95%-90%-80%) and the resulting orange/yellow solid was then stirred in DCM:MeOH (90%, 20 mL) when polymer supported trialkylammonium mesylate resin (600 mg) was added. After 1 h, the reaction mixture was filtered, concentrated in vacuo and the residue triturated in Et20/hexanes (20 mL) to yield a red hygroscopic solid. This solid was then redissolved in MeOH:DCM (10%, 20mL) when trialkylammonium formate resin (300 mg) was added. After 2 h the mixture was filtered and the filtrate concentrated in vacuo and then absorbed onto silica and purified by flash chromatography (gradient elution DCM:MeOH 100%-95%-90%-80%) to give the title compound as an orange solid (12 mg, 0.03 mmol, 22% yield).
<¾ (DMSO-dg): 8.87-8.90 (1H, d, J=9.2), 8.64-8.66 (1H, d, J=7.9), 8.56-8.58 (1H, dd, J=8.4, 1.0), 8.34-8.38 (1H, t, J=8.1), 8.06-8.10 (1H, ddd, J=8.4, 7.2, 1.2), 7.88-7.91 (2H, m), 7.79-7.80 (1H, d, J=2.3), 7.77 (1H, br s), 7.68-7.71 (1H, ddd, J=8.3, 7.0, 1.0), 7.60-7.62 (1H, dd, 9.1, 2.2), 7.56 (1H, br s), 4.84 (3H, s), 4.60 (3H, s).
m/z (ES+): 357.0 (M+).
Example 63 - Preparation of l,ll-difluoro-2-hYdroxy-13-methylchromenor4,3.i2- g/¾lphenanthridin-13-ium bromide
Figure imgf000141_0001
A 1M solution of boron tribromide in dichloromethane (1 mL) was added to a suspension of 1 , 1 l-difluoro-2-methoxy- 13-methylchromeno[4,3 ,2-g¾phenanthridin- 13-ium
trifluoromefhanesulfonate (38 mg, 0.076 mmol; Example 17) in DCM (2 mL) and the mixture was stirred at 25 °C. After 16 h, the reaction was quenched with MeOH and the mixture was stirred for ~30min, then concentrated in vacuo. The crude material was then absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 96%-92%) and the resulting red solid was sonicated in Et20/MeOH, then filtered and washed Et20 and air dried to give the title compound as a dark red solid (32 mg, 0.076 mmol, 100% yield).
4 (DMSO-d6): 11.48 (1H, s), 8.56-8.61 (3H, m), 8.38-8.42 (1H, t, J=8.2), 7.99-8.08 (2H, m), 7.92-7.94 (1H, d, J=8.2), 7.52-7.77 (1H, t, J=8.0), 4.59-4.61 (3H, d, J=8.2).
m/z (ES+): 336.0 (M+). Example 64 - Preparation of 3,ll-difluoro-2-(isopropoxycarbonyl)-13- methylchromenor4,3,2-gfelphenanthridin-13-ium chloride
Figure imgf000142_0001
A solution of 3,l l-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium trifluoromethane sulfonate (59 mg, 0.112 mmol; Example 49) in 6N hydrochloric acid (10 mL) was heated at 90°C. After 4 h, the reaction mixture was cooled and evaporated in vacuo to give 2-carboxy-3,l l-difluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium chloride (38 mg, 0.095 mmol, 85% yield) (m/z (ES+): 364.0 (MH+). The material was used without further characterization and purification in the next steps.
A suspension of 2-carboxy-3,l l-difluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium chloride (38 mg, 0.095 mmol) in phosphorous oxycloride (10 mL) was refluxed for 2 h then allowed to cool to room temperature. The mixture was concentrated in vacuo and the product was co-evaporated with toluene and hexane to give 2-(chlorocarbonyl)-3,l l-difluoro-13- methylchromeno[4,3,2-g/z]phenanthridin-13-ium chloride as a brown/green solid. The material was used without further characterization and purification in the next steps.
A suspension of crude 2-(chlorocarbonyl)-3,l l-difluoro-13-methylchromeno[4,3,2- g/j]phenanthridin-13-ium chloride in anhydrous pyridine (2 mL) and propan-2-ol (2 mL) was stirred at room temperature. After 2 h, the reaction mixture was concentrated in vacuo, and the resulting solid was azeotroped with toluene. The crude material was absorbed on silica, and purified by flash chromatography (gradient elution DCM:MeOH 94%-90%). The resulting yellow solid was then purified by CI 8 reverse-phase chromatography (gradient elution watenmethanol 50%-100%) to give the title compound as a red solid (18 mg, 0.041 mmol, 37% yield).
<¾ (DMSO-d6): 8.96-8.99 (1H, d, J=11.4), 8.84-8.86 (1H, d, J=7.7), 8.66-8.68 (1H, d, J=6.2), 8.48-8.52 (1H, t, J=8.2), 8.39-8.42 (1H, dd, J=9.7, 2.4), 8.13-8.16 (1H, d, J=8.1), 8.03-8.09 (2H, m), 5.28-5.34 (1H, sept, J=6.2), 4.71 (3H, s), 1.42-1.44 (6H, d, J=6.2).
m/z (ES+): 406.0 (M+). Example 65 - 2-(methoxymethoxy)-13-methylchromeno[4,3,2-gMphenanthridin-13-ium chloride
Figure imgf000143_0001
To a solution of 2-hydroxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (20 mg, 0.045 mmol; Example 6) in dichloromethane (2 mL) was added diisopropylethylamine (0.118 mL, 0.445 mmol, 10 equ) and chloromethyl methyl ether (0.050 mL, 0.445 mmol, 10 equ) and the reaction was stirred at 25°C. After 15 mins further amounts of 2-hydroxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate (40 mg, 0.089 mmol, 2 equ), diisopropylethylamine (0.118 mL, 0.445 mmol, 10 equ) and chloromethyl methyl ether (0.050 mL, 0.445 mmol, 10 equ) were added and the RM stirred for a further 0.5 h when the reaction mixture was then taken up in MeOH:DCM (10%), filtered with MeOH washings. The filtrate was then absorbed onto silica and was purified by flash chromatography (gradient elution DCM:MeOH 100%-95%-90%-80%) and the resulting orange/red solid was then stirred in DCM:MeOH (90%, 20 mL) when silica supported trialkylammonium chloride resin (600 mg) was added. After 1 h, the reaction mixture was filtered, concentrated in vacuo and the residue triturated in Et20 hexanes (20 mL) to give the title compound as an orange solid (16 mg, 0.036 mmol, 27% yield).
<¾ (DMSO-d6): 8.87-8.90 (1H, d, J=9.2), 8.65-8.67 (1H, d, J=7.8), 8.55-8.57 (1H, dd, J=8.5, 1.2), 8.35-8.39 (1H, t, J=8.2), 8.06-8.08 (1H, ddd, J=8.5, 7.4, 1.3), 7.87-7.93 (3H, m), 7.67-7.72 (1H, ddd, J=8.4, 7.2, 1.2), 7.64-7.67 (1H, dd, 9.0, 2.2), 5.56 (2H, s), 4.60 (3H, s), 3.49 (3H, s) m/z (ES+): 344.0 (M+). Example 66 - Preparation of 2-((2-hvdroxyethoxy)carbonvI)-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000144_0001
Molecular Weight: 521 .46
A mixture of 2-carboxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (100 mg, 0.25 mmol; Example 51) in phosphorus oxychloride (2 mL) was heated at 100°C for 1 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-13- methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate, which was used without further purification in the next step (quantitative yield assumed). Crude 2- (chlorocarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate was dissolved in chloroform (20 mL) and ethylene glycol (0.068mL, 1.2 mmol, 5 equ) and the resulting solution was stirred at 25°C. After 2 h, the mixture was concentrated in vacuo and the crude material was absorbed on silica, and purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a yellow solid (20 mg, 0.038 mmol, 16% yield).
<¼ (DMSO-d6): 9.06-9.08 (1H, d, J=8.6), 8.84-8.86 (1H, d, J=7.8), 8.79-8.79 (1H, d, J=1.3), 8.59-8.61 (1H, dd, J=8.6, 1.0), 8.47-8.51 (1H, t, J=8.2), 8.43-8.45 (1H, dd, J=8.5, 1.3), 8.12-8.16 (2H, m), 7.95-7.98 (1H, dd, J=8.4, 1.0), 7.74-7.78 (1H, ddd, J=8.4, 7.2, 1.1), 5.05-5.08 (1H, t, J=5.8), 4.71 (3H, s), 4.45-4.47 (2H, t, J=4.8), 3.80-3.84 (2H, q, J=5.8).
Example 67 - Preparation of ll-fluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- g¾lphenanthridin-13-ium trifluoromethanesulfonate:
Step 1 - Preparation of 3-(2-bromo-4-fluorophenoxy)-2'-fluoro-4'-methoxybiphenyl-2- carbonitrile
Figure imgf000144_0002
A mixture of potassium carbonate (6.6 g, 47 mmol, 2 equ), 2',3-difluoro-4'-methoxybiphenyl-2- carbonitrile (5.8 g, 24 mmol, Example 4) and 2-bromo-4-fluorophenol (1.4 g, 7.3 mmol, 2 equ) in N,N-dimethylformamide (12 mL) was heated at 100°C for 44 hrs and was allowed to cool to room temperature. Saturated ammonium chloride solution (20 mL) was added and the aqueous layer extracted with diethyl ether (3 x 100 mL). The combined organic extracts were concentrated in vacuo and the residue was purified by flash chromatography (chloroform) to give the title compound as a white solid (4.3 g, 44% yield).
<¾ (CDC13): 7.47-7.50 (1H, dd, J=8.5, 7.7), 7.43-7.46 (1H, dd, J=7.7, 2.6), 7.38-7.42 (1H, d, J=8.6), 7.11-7.20 (3H, m), 6.83-6.85 (1H, ddd, J=8.6, 2.6, 0.6), 6.77-6.81 (1H, dd, J=11.9, 2.4), 6.61-6.63 (1H, dd, J=8.5, 0.9), 3.68 (3H, s).
Sp (CDCI3): -112.6 (IF, dd, J=11.7, 8.6), -114.3 (IF, m).
Step 2 - Preparation of ll-fluoro-2-methoxychromeno[4,3,2-^A]phenanthridine
Figure imgf000145_0001
Tert-butyl lithium (1.6M in pentane, 3.1 mL, 5.0 mmol, 2.1 equ) was added to anhydrous tetrahydrofuran (10 mL) at -78°C and the mixture was stirred for 5 mins. 3-(2-bromo-4- fluorophenoxy)-2'-fluoro-4'-methoxybiphenyl-2-carbonitrile (1.0 g, 2.4 mmol) dissolved in anhydrous tetrahydrofuran (15 mL) was added drop wise. The resulting deep red solution was stirred at -78°C for 5 min and the reaction was stirred at room temperature for 15 mins. A saturated solution of ammonium chloride (10 mL) was added and the mixture was extracted with dichloromethane (2 x 20 mL). The combined organic extracts were concentrated in vacuo and the residue was purified by flash chromatography (gradient elution chloroform: dichloromethane) to give the title compound as a pale yellow solid (1.65 g, 50% yield).
<¼ (CDCI3): 8.36 (1H, m), 8.30-8.33 (1H, d, J=9.0), 8.06-8.08 (1H, d, J=7.7), 7.82 (1H, m), 7.54 (1H, m), 7.20-7.32 (4H, m), 4.03 (3H, s).
Step 3 - Preparation of methyl ll-fluorochromeno[4,3,2-gft]phenanthridine-2-carboxylate
Figure imgf000145_0002
A mixture of l l-fluoro-2-methoxychromeno[4,3,2-g¾]phenanthridine (1.65 g, 5.2 mmol) and pyridine hydrochloride (6 g, 52 mmol, 10 equ) was heated at 210°C for 2 h and was allowed to cool to room temperature. Water (20 mL) was added and the resulting precipitate was filtered, washed with water and dried under suction to give l l-fluorochromeno[4,3,2-g z]phenanthridin-2- ol (1.55 g, 98% yield). The material was used without further purification and characterization in the next step.
A mixture of crude l l-fluorochromeno[4,3,2-gA]phenanthridin-2-ol (1.5 g, 5.0 mmol), N- Phenyl-bis(trifluoromethanesulfonimide) (3.5 g, 9.9 mmol, 2 equ) and Ν,Ν- diisopropylethylamine (1.3 g, 9.9 mmol, 2 equ) in 1:1 chloroform:tetrahydrofuran (50 mL) was heated at 70°C for 72 h and was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo and the residue was purified by flash chromatography (gradient elution Hexane:CHCi3 50%-80%) to give l l-fluorochromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate as a cream solid (1.84 g, 85% yield). The material was used without further characterization in the next step.
A mixture of l l-fluorochromeno[4,3,2-g/i]phenanthridin-2-yl trifluoromethanesulfonate (200 mg, 0.57 mmol), palladium acetate (2.5 mg, 0.011 mmol, 0.02 equ), 4,5-bis(diphenylphosphino)- 9,9-dimethylxanthene (13 mg, 0.022 mmol, 0.04 equ) and triethylamine (116 mg, 1.15 mmol, 2 equ) in 2: 1 N,N-dimethylformamide:methanol (7.5 mL) was heated at 65°C under a carbon monoxide atmosphere for 2 h and was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo and the residue was purified by flash chromatography (dichloromethane) to give the title compound as a white solid (39 mg, 20% yield).
<¾ (DMSO-ifc): 8.78-8.80 (1H, d, J=8.3), 8.56 (1H, s (br)), 8.50-8.52 (1H, d, J=8.0), 8.23-8.25 (1H, d, J=8.8), 8.10-8.13 (1H, d, J=8.4), 8.01-8.06 (1H, dd, J1=J2=8.0), 7.62-7.64 (1H, d, J=8.0), 7.57-7.58 (2H, m), 3.97 (3H, s).
Step 4 - Preparation of ll-fluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- g/i]phenanthridin-1 -ium trifluoromethanesulfonate
Figure imgf000146_0001
A mixture of methyl l l-fluorochromeno[4,3,2-g¾phenanthridine-2-carboxylate (100 mg, 0.29 mmol), potassium carbonate (60 mg, 0.43 mmol, 1.5 equ) and methyl triflate (230 mg, 1.4 mmol, 5 equ) was heated at 120°C in a sealed tube for 16 h and was allowed to cool to room temperature. The inorganic materials were removed by filtration and washed with 90% DCM:MeOH. The filtrate was concentrated in vacuo and the residue was purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a yellow solid (15 mg, 10%).
<¼ (DMSO-i¾): 9.06-9.09 (1H, d, J=8.6), 8.84-8.87 (1H, d, J=7.8), 8.74-8.75 (1H, d, J=1.5), 8.48-8.52 (1H, dd, J1=J2=8.2), 8.39-8.43 (2H, m), 8.12-8.14 (1H, d, J=7.9), 8.04-8.10 (2H, m), 4.73 (3H, s), 4.04 (3H, s).
Example 68 - Preparation of 2-carbamovI-4,ll-difluoro-13-methylchromeno[4,3,2- g/¾lphenanthridin-13-ium methanesulfonate
Step 1 - Preparation of 2-carboxy-4,ll-difluoro-13-methylchromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000147_0001
[00185] A solution of 4,l l-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- g z]phenanthridin-13-ium trifluoromethanesulfonate (47 mg, 0.085 mmol; Example 57) in 6M hydrochloric acid (4 mL) was heated at 90 °C for 4 h. The RM was allowed to cool and evaporated to dryness in vacuo then co-evaporated with toluene to give 2-carboxy-4,l l-difluoro-
13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate as a yellow solid that was assumed 100% yield used without further purification.
[00186]m/z (ES+): 364.0(M+).
Step 2 - Preparation of 2-carbamoyl-4,ll-difluoro-13-methylchromeno[4,3,2- g¾]phenanthridin-13-ium methanesulfonate
Figure imgf000147_0002
[00187]A mixture of 2-carboxy-4,l l-difluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13- ium trifluoromethanesulfonate (46 mg, 0.085 mmol) in phosphorus oxychloride (4 mL) was heated at 90°C for 4 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-4,ll-difluoro-13- methylchromeno[4,3,2- z]phenanthridin-13-trifluoromethanesulfonate, which was used without further purification in the next step (quantitative yield assumed). Crude 2-(chlorocarbonyl)-4,ll- difluoro- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-trifluoromethanesulfonate was dissolved in tetrahydrofuran (4 mL) and the resulting solution was added to liquid ammonia in tetrahydrofuran (5 mL) at 0°C. The mixture was allowed to warm to 25°C over 1 h and then concentrated to dryness in vacuo. The residue was purified by flash chromatography (gradient elution DCM:MeOH 90%-85%) and the resulting solid was redissolved in 10% dichloromethane: methanol (2 mL) when Amberlyst resin, mesylate form (200 mg) was added. The mixture was stirred for 2 h then filtered and the filtrate was concentrated in vacuo to give the title compound as an orange solid (4.5 mg, 8% yield).
[00188] 4 (DMSO-t½): 8.79-8.81 (1H, d, J=8.9), 8.57-8.59 (2H, m), 8.50-8.59 (1H, t, J=7.9), 8.41-8.44 (1H, dd, J=9.8, 2.5), 8.25-8.28 (1H, d, J=13.2), 8.16-8.18 (1H, d, J=8.8), 8.03-8.11 (3H, m), 4.68 (3H, s), 2.29 (3H, s).
Example 69 - Preparation of ll-fluoro-2-(methoxymethyl)-13-methylchromeno[4,3,2- g½1phenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of (ll-fluorochromeno[4,3,2-^]phenanthridin-2-yl)methanol
Figure imgf000148_0001
[00189]To a solution of l l-fluorochromeno[4,3,2- z]phenanthridine-2-carboxylate (100 mg, 0.29 mmol, Example 67) in dichloromethane (10 mL) was added diisobutylaluminium hydride 1M in dichloromethane (1.4 mL, 1.4 mmol, 5 equ) and stirred for 2 h. The reaction mixture was diluted with methanol then concentrated to dryness in vacuo. The product was purified by flash chromatography (DCM:MeOH 95%) to give the title compound as a pale yellow solid (57 mg, 62% yield).
<¾ (DMSO-d6): 8.61-8.63 (1H, d, J=7.9), 8.41-8.43 (1H, d, J=7.9), 8.20-8.23 (1H, dt, J+8.6, 1.9), 7.99- 8.00 (1H, d, J=1.3), 7.94-7.98 (1H, t, J=8.1), 7.60-7.62 (1H, dd, J=8.4, 1.7), 7.55-7.56 (1H, d, J=1.8), 7.49-7.53 (2H, m), 5.43-5.46 (1H, t, J=5.8), 4.74-4.76 (2H, d, J=5.8). Step 2 - Preparation of ll-fluoro-2-(methoxymethyl)-13-methylchromeno[4,3,2- g/i]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000149_0001
[00190]A mixture of (l l-fluorochromeno[4,3,2-g/i]phenanthridin-2-yl)methanol (185 mg, 0.6 mmol), potassium carbonate (203 mg, 1.6 mmol, 2.5 equ) and methyl triflate (480 mg, 2.9 mmol, 5 equ) was heated at 120°C in a sealed tube for 48 h and then allowed to cool to room temperature. The inorganic materials were removed by filtration and washed with 80% DCM:MeOH. The filtrate was concentrated in vacuo and the residue was purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a yellow solid (150 mg, 52% yield).
<¾ (DMSO-cfc):8.91-8.94 (1H, d, J=8.4), 8.75-8.77 (1H, d, J=7.8), 8.39-8.45 (2H, m), 8.28 (1H, s (br)), 7.98-8.07 (3H, m), 7.88-7.91 (1H, dd, J=8.3, 1.0), 4.78 (2H, s), 4.66 (3H, s), 3.46 (3H, s).
m/z (ES+): 346.0 (M+). Example 70 - Preparation of 4,ll-difluoro-13-methyl-2-(lH-pyrazol-4-Yl)chromeno[4,3<2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of 4,ll-difluoro-13-methyl-2- (trifluoromethylsulfonyloxy)chromeno[4,3,2-^¾]phenanthridin-13-ium
trifluoromethanesulfonate
Figure imgf000149_0002
[00191] Methyl triflate (242 μL·, 2.2 mmol, 10 equ) was added to a suspension of 4,11- difluorochromeno[4,3,2-g/z]phenanthridin-2-yl trifluoromethanesulfonate (100 mg, 0.22 mmol, Example 57) and potassium carbonate (464 mg, 2.2 mmol, 10 equ) in anhydrous chloroform (10 mL) and the mixture was heated at 110°C for 5 h in a sealed tube. An additional 10 equivalents of methyl triflate were added and the mixture was heated at 100°C in a sealed tube overnight. The reaction mixture was cooled to room temperature and was quenched with methanol. The mixture was filtered and the filtrate was transferred in a separating funnel. Water was added and the product was extracted with dichloromethane. The combined organic extracts were concentrated in vacuo and the product (assumed quantitative yield) was used without further purification in the next step.
<¼ (DMSO-d6): 8.76-8.78 (1H, d, J=8.2), 8.50-8.53 (2H, m), 8.45-8.48 (1H, dd, J=9.8, 2.8), 8.32- 8.36 (1H, dd, J=12.3, 2.3), 8.19-8.21 (1H, dd, J=8.4, 0.8), 8.07-8.15 (2H, m), 4.50 (3H, s).
m/z (ES+): 467.9 (M+).
Step 2 - Preparation of 4,ll-difluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000150_0001
[00192]A mixture of crude 4,ll-difluoro-13-methyl-2-
(trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium triflate (100 mg, 0.162 mmol), lH-pyrazole-4-boronic acid pinacol ester (62 mg, 0.32 mmol, 2 equ), tetrakis(triphenylphosphine)palladium(0) (25 mg, 0.0213 mmol, 0.13 equ) and sodium acetate (38 mg, 0.469 mmol, 2.9 equ) in DME:H20 3:1 (1.5 mL) was heated under microwave radiation for 20min. at 100°C (300W, 200psi, cooling system on, ramp time 30s). 1.5 equ of lH-pyrazole- 4-boronic acid pinacol ester and 0.1 equ of tetrakis(triphenylphosphine)palladium(0) were added and the mixture was re-heated under microwave radiation for 20 min. at 100°C (300W, 200psi, cooling system on, ramp time 30s). An additional 0.1 equ of tetrakis(triphenylphosphine)palladium(0) were added and the mixture was heated under microwave radiation for 15 min. (300W, 200psi, cooling system on, ramp time 30s). 1.5 equ 1H- pyrazole-4-boronic acid pinacol ester and 0.1 equ tetrakis(triphenylphosphine)palladium(0) were added and the mixture was heated under microwave radiation for 20 min. at 100°C (300W, 200psi, cooling system on, ramp time 30s). The mixture was concentrated to dryness in vacuo and the crude mixture was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-96%-94%-92 ).The resulting solid was triturated with 3N HC1 and the precipitate was filtered, washed with diethyl ether and dried under suction to give the title compound as bright yellow/orange solid (7 mg, 8% yield) [00193] <¾ (DMSO- ): 13.3 (1H, s), 8.69-8.72 (2H, m), 8.42-8.47 (1H, dd, J1=J2=8.1), 8.37-8.40 (2H, m), 8.30 (1H, s (br)), 8.21-8.24 (1H, dd, J=14.4, 1.0), 8.01-8.07 (3H, m), 4.68 (3H, s).
[00194] Sp (DMSO-Je): -77.8, -109.6 (d, J=13.9), -114.5 (m).
[00195] m/z (ES+): 386.0 (M+).
Example 71 - Preparation of 4,ll-difluoro-13-methyl-2-((tetrahvdrofuran-2- yloxy)carbonyl)chromeno[4,3,2-jgA]phenanthridin-13-ium chloride
Figure imgf000151_0001
[00196]A mixture of 2-carboxy-4,l l-difluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13- ium trifluoromethanesulfonate (70 mg, 0.175 mmol, Example 68) in phosphorus oxychloride (5 mL) was heated at 100°C for 2 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-4,l l- difluoro-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium chloride, which was used without further purification in the next step (quantitative yield assumed). Crude 2-(chlorocarbonyl)-4,l l- difluoro-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium chloride was dissolved in pyridine (3 mL) when 3-hydroxytetrahydrofuran (154 mg, 1.75 mmol, 10 equ) was added in one portion. The mixture was stirred at 0°C for 0.5 h and was then concentrated to dryness in vacuo. The product was purified by flash chromatography (gradient elution DCM:MeOH 92.5%-90%) to give the title compound as a yellow solid (30 mg, 35% yield).
4 (DMSO-d6): 8.82-8.84 (1H, d, J=8.3), 8.52-8.55 (2H, m), 8.41-8.44 (1H, dd, J=9.7, 2.6), 8.28- 8.31 (1H, dd, J=12.6, 1.2), 8.21-8.23 (1H, d, J=8.4), 8.07-8.13 (2H, m), 5.61-5.64 (1H, m), 4.70 (3H, s), 3.94-4.03 (3H, m), 3.82-3.87 (1H, dt, J=8.4, 4.7), 2.29-2.36 (1H, m), 2.17-2.23 (1H, m). δρ {OMSO-de): -77.8, -107.6 (d, J=12.6), -114.2 (m)
m/z (ES+): 434.0 (M+). Example 72 - Preparation of 4 Jl-difluoro-2-((3-hvdroxypropoxy)carbonyl)-13- methylchromenor4,3,2-gfelphenanthridin-13-ium chloride
Figure imgf000152_0001
[00197]A mixture of 2-carboxy-4,l l-difluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13- ium trifluoromethanesulfonate (40 mg, 0.100 mmol, Example 68) in phosphorus oxychloride (10 mL) was heated at 100°C for 0.5 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-4,l l- difluoro-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium chloride, which was used without further purification in the next step (quantitative yield assumed). Crude 2-(chlorocarbonyl)-4,l l- difluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium chloride was dissolved in pyridine (2 mL) when 1,3-propanediol (1 mL) was added in one portion. The mixture was stirred at -20°C for 0.5 h and was then concentrated to dryness in vacuo. The product was purified by flash chromatography (gradient elution DCM:MeOH 99%-95%-93%-90%) and the resulting solid was triturated with diethyl ether and filtered to give the title compound as a yellow solid (5 mg, 11% yield).
<¾ (DMSO-d6): 8.82-8.84 (1H, d, J=8.3), 8.52-8.57 (2H, m), 8.41-8.44 (1H, dd, J=9.6, 2.5), 8.25- 8.28 (1H, dd, J=12.6, 1.2), 8.20-8.22 (1H, dd, J=8.5, 0.6), 8.06-8.14 (2H, m), 4.70 (3H, s), 4.64- 4.67 (1H, t, J=5.2), 4.49-4.53 (2H, t, J=6.5), 3.62-3.66 (2H, dd, J=6.2, 5.2), 1.93-2.00 (2H, quin, J=6.1).
mix (ES+): 422.1 (M+).
Example 73 - Preparation of 4,ll-difluoro-2-((2-methoxyethoxy)carbonyl)-13- methylchromenor4,3,2-gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000152_0002
[00198]A mixture of 2-carboxy-4,l l-difluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13- ium trifluoromethanesulfonate (60 mg, 0.150 mmol, Example 68) in phosphorus oxychloride (5 mL) was heated at 100 C for 2 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chlorocarbonyl)-4,l l- difluoro- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium trifluoromethanesulfonate, which was used without further purification in the next step (quantitative yield assumed). Crude 2 2- (chlorocarbonyl)-4, 11 -difluoro- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium
trifluoromethanesulfonate was dissolved in pyridine (2 mL) when 2-methoxyethanol (114 mg, 1.50 mmol, 10 equ) was added in one portion. The mixture was stirred for 0.5 h and was then concentrated to dryness in vacuo. The product was purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) and the resulting solid was triturated with diethyl ether and filtered to give the title compound as a yellow solid (22 mg, 26% yield).
<¼ (DMSO-d6): 8.82-8.84 (1H, d, J=8.3), 8.52-8.56 (2H, m), 8.42-8.45 (1H, dd, J=9.5, 2.2), 8.24- 8.27 (1H, dd, J=12.6, 1.2), 8.20-8.23 (1H, dd, J=8.4, 0.4), 8.08-8.12 (2H, m), 4.70 (3H, s), 4.57- 4.59 (2H, m), 3.76-3.78 (2H, m), 3.36 (3H, s).
Sp (DMSO-<4): -77.6, -107.4 (d, J=12.5), -114.1 (m).
m/z (ES+): 422.0 (M+).
Example 74 - Preparation of ll-fluoro-13-methyl-2-(morpholinomethyl)chromeno[4,3,2- g¾lphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of 2-(bromomethyl)-ll-fluoro-13-methylchromeno[4,3,2- ^]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000153_0001
[00199]A 1M solution of boron tribromide in dichloromethane (1.5 mL, 1.5 mmol, 5.0 equ) was added dropwise to a suspension of l l-fluoro-2-(methoxymethyl)-13-methylchromeno[4,3,2- g/j]phenanthridin-13-ium trifluoromethanesulfonate (150 mg, 0.3 mmol, Example 69) in dichloromethane (5 mL) at 25°C. After 3 h, the reaction was quenched with a few drops of methanol then concentrated to dryness in vacuo. The residue was purified by flash chromatography (DCM:MeOH 90%) to give the title compound as a yellow solid (120 mg, 73% yield).
<¾ (DMSO-i¾: 8.94-8.98 (1H, t, J=8.6), 8.76-8.79 (1H, dd, J=7.8, 3.4), 8.51-8.53 (1H, dd, J=8.6, 1.0), 8.43-8.47 (2H, m), 8.01-8.06 (4H, m), 5.07-5.14 (2H, d, J=28.0), 4.65-4.66 (3H, d, J=4.1). m/z (ES+): 394.0 (M+, 79Br), 395.9 (M+, 81Br).
Step 2 - Preparation of ll-fluoro-13-methyl-2-(morpholinomethyl)chromeno[4,3,2- g¾]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000154_0001
[00200]To a solution of 2-(bromomethyl)-l l-fluoro-13-methylchromeno[4,3,2- g/z]phenanthridin-13-ium trifluoromethanesulfonate (35 mg, 0.06 mmol) in tetrahydrofuran (3 mL) was added morpholine (28 mg, 0.32 mmol, 5.0 equ) and the resulting solution was stirred for 16 h at 25°C. After 6 h, sodium carbonate was added and stirred for a further 2 h then concentrated to dryness in vacuo. The residue was purified by flash chromatography (DCM:MeOH 90%) to give the title compound as a yellow solid (15 mg, 42% yield).
4 (CD3OD): 8.80-8.83 (1H, d, J=8.4), 8.67-8.69 (1H, d, J=8.0), 8.38-8.42 (1H, dd, J1=J2=8.1), 8.32-8.35 (2H, m), 7.90-7.96 (4H, m), 4.76 (3H, s), 3.87 (2H, s), 3.75-3.77 (4H, m), 2.58-2.61 (4H, m).
δρ (CD3OD): -80.1, -115.8
m/z (ES+): 401.0 (M+).
Example 75 - Preparation of ll-fluoro-13-methyl-2-(3-methyl-l,2,4-oxadiazoI-5- yl)chromeno[4,3,2-gAlphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000154_0002
[00201]A mixture of 2-carboxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (50 mg, 0.112 mmol; Example 51) in phosphorus oxychloride (3 mL) was heated at 100°C for 1 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo then co-evaporated with anhydrous toluene to give 2-(chlorocarbonyl)- 13-methylchromeno[4,3,2-g/?]phenanthridin- 13-ium trifluoromethanesulfonate. Crude 2-(chlorocarbonyl)-13-methylchromeno[4,3,2- g j]phenanthridin-13-ium trifluoromethanesulfonate was diluted with anhydrous dichloromethane (3 mL) and triethylamine (36 μί, 0.125 mmol, 1.1 equ) followed by the addition of acetamide oxime (19 mg, 0.125 mmol, 1.1 equ) and the reaction was stirred under an inert atmosphere. After 2 hours, the reaction mixture was diluted with anhydrous toluene, a condenser was fitted and the reaction was heated to 110°C for a further 2 hours. After cooling the solution was concentrated to dryness in vacuo and the product was purified by flash chromatography (DCM:MeOH 95%-90%) to give the title compound as a yellow solid (15 mg, 28% yield).
<¾ (DMSO-dg): 9.05-9.16 (1H, d, J=8.6), 8.90-8.91 (1H, d, J=1.4), 8.83-8.88 (1H, d, J=7.7), 8.59-8.60 (1H, dd, J=8.5, 1.3), 8.53-8.56 (1H, dd, J=8.5, 1.4), 8.48-8.52 (1H, t, J=8.1), 8.11-8.17 (2H, m), 7.96-7.99 (1H, dd, J=8.5, 1.1), 7.75-7.79 (1H, dt, J=8.4, 1.1), 4.74 (3H, s), 3.30 (3H, s). m/z (ES+): 366.0 (M+). Example 76 - Preparation of 2-((dimethylamino)methyl)-ll-fluoro-13- methylchromenor4,3,2-gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000155_0001
[00202]A 2M solution of dimethylamine in tetrahydrofuran (0.137 mL, 0.27 mmol, 5.0 equ) was added to a solution of 2-(bromomethyl)-ll-fluoro-13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium trifluoromethanesulfonate (30 mg, 0.05 mmol, Example 74) in tetrahydrofuran (1 mL) at 25°C. The resulting solution was stirred for 1 h then filtered and concentrated to dryness in vacuo. The residue was purified by flash chromatography (gradient elution DCM:MeOH 90%- 80%) to give the title compound as a yellow solid (8 mg, 28% yield).
<¾ (CD3OD): 8.85-8.87 (1H, d, J=8.4), 8.69-8.71 (1H, d, J=7.8), 8.39-8.43 (1H, dd, J1=J2=8.1), 8.33-8.36 (2H, m), 7.95-7.97 (1H, dd, J=8.3, 0.7), 7.89-7.98 (3H, m), 4.70 (3H, s), 3.92 (2H, s, (br)), 2.46 (6H, s).
* (CD3OD): -82.6, -118.1
m/z (ES+): 346.0 (M+). Example 77 - Preparation of (2-carboxy-ll-fluoro-13-methylchromeno[4,3,2- gfelphenanthridin-13-ium).(2,2,2-trifluoroacetate).(trifluoromethanesuIfonate)
Figure imgf000156_0001
[00203]A solution of ll-fluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2- g/z]phenanthridin-13-ium trifluoromethanesulfonate (150 mg, 0.3 mmol, Example 67) in trifluoroacetic acid (5 mL) and water (1 mL) was stirred at 120°C for 16 h then concentrated to dryness in vacuo. The product was purified by flash chromatography (gradient elution DCM:MeOH 90%-75 ) to give the title compound as a yellow solid (85 mg, 58% yield) as a mixture of trifluoromethanesulfonate and 2,2,2-trifluoroacetate salts.
[00204] <¾ (DMSO-d6): 8.90-8.92 (1H, d, J=8.5), 8.77-8.79 (1H, d, J=8.0), 8.73-8.77 (1H, s), 8.41-8.46 (2H, m), 8.36-8.38 (1H, d, J=8.3), 8.00-8.05 (3H, m), 7.12 (1H, s (br)), 4.71 (3H, s).
[00205] m/z (ES+): 346.0 (M+).
Example 78 - Preparation of 2-(2-amino-2-oxoethoxy)-4,ll-difluoro-13- methylchromenor4,3,2-gfe]phenanthridin-13-ium methanesulfonate
Figure imgf000156_0002
[00206] Methyl triflate was added to a suspension of l,ll-difluoro-2-methoxychromeno[4,3,2- gA]phenanthridine (64 mg, 0.191 mmol, Example 29) and potassium carbonate (264 mg, 1.91 mmol, 10 equ) in chloroform (3 mL) and the mixture was heated in a sealed tube at 110°C. After 16 h, the reaction mixture was cooled to room temperature and quenched with MeOH. The mixture was stirred for 0.5 h and the inorganic salts were removed by filtration and washed with 90% DCM:MeOH. The filtrate was transferred into a separating funnel and water was added. The product was extracted with DCM and the combined organic extracts were dried over magnesium sulfate and concentrated to dryness in vacuo to give 4,l l-difluoro-2-methoxy-13- methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate. The product was used without further purification in the next step.
[00207]A 1M solution of boron tribromide (175 μί, 0.175 mmol, 3.5 equ) was added to a suspension of 4,1 l-difluoro-2-methoxy-13-methylchromeno[4,3,2-g ?]phenanthridin-13-ium trifluoromethanesulfonate (25 mg, 0.050 mmol) in dichloromethane (2 mL) and the mixture was stirred for 2 h. The reaction was quenched with methanol and the resulting solution was concentrated in vacuo to give 4,l l-difluoro-2-hydroxy-13-methylchromeno[4,3,2- g/j]phenanthridin-13-ium bromide as a dark purple solid (quantitative yield assumed) (m/z (ES+): 336.0 (M+). The product was used without further characterization and purification in the next step.
[00208]A mixture of crude 4,l l-difluoro-2-hydroxy-13-methylchromeno[4,3,2- g/z]phenanthridin-13-ium bromide (25 mg, 0.060 mmol) and potassium carbonate (83 mg, 0.60 mmol, 10 equ) in anhydrous Ν,Ν-dimethylformamide (1 mL) was stirred for 5 minutes before the addition of iodoacetamide (61 mg, 0.60 mmol, 10 equ). The mixture was heated at 60°C for 1 h and then allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo and the crude product was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-96%-94%-92%-90%) to give 22mg of a yellow/orange solid. The product was dissolved in DCM:MeOH and amberlyst A- 24 mesylate resin (330 mg) was added. The mixture was stirred for 1 h and was filtered. The filtrate was concentrated to dryness in vacuo and the resulting solid was triturated with 10% MeOH:Et20. The resulting precipitate was filtered to give 14 mg of an orange solid. The product was dissolved in water and tetrahydrofuran was added. The resulting precipitate was filtered, washed with diethyl ether and dried under suction to give the title compound as an orange solid (6 mg, 21% yield).
[00209] Note: NMR analysis of this material showed that it was 70% pure and was contaminated with 30% wt. potassium mesylate.
[00210] The filtrate was concentrated in vacuo and 5% aq. dioxane was added. The mixture was kept at 4°C for 16 h and the resulting precipitate was filtered, washed with diethyl ether and dried under suction to give the pure title compound as a yellow/orange solid (2 mg, 7% yield).
[00211] <¾ (DMSO-i¾): 8.62-8.65 (1H, d, J=8.8), 8.40-8.45 (2H, m), 8.00-8.07 (3H, m), 7.78 (1H, m), 7.68 (1H, m), 7.61-7.65 (1H, m), 7.58-7.59 (1H, m), 4.85 (2H, s), 4.58 (3H, s, ), 2.28 (3H, s).
[00212]m z (ES+): 393.0 (M+). Example 79 - Preparation of 2-carbamovI-ll-fluoro-13-methylchromeno[4,3^2- gfelphenanthridin-13-ium methanesulfonate
Figure imgf000158_0001
[00213]A mixture of (2-carboxy-l l-fluoro-13-methylchromeno[4,3,2- /z]phenanthridin-13-ium) 0.5(2,2,2-trifluoroacetate) 0.5(trifluoromethanesulfonate) (85 mg, 0.17 mmol; Example 77) in phosphorus oxychloride (2 mL) was heated at 100°C for 3 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo to give 2-(chloroc arbonyl) - 11 -fluoro- 13 -methylchromeno [4,3 ,2-gA]phenanthridin- 13 -ium
trifluoromethanesulfonate, which was used without further purification in the next step (quantitative yield assumed). Crude (2-(chlorocarbonyl)-l l-fluoro-13-methylchromeno[4,3,2- g i]phenanthridin-13-ium)0.5(2,2,2-trifluoroacetate) 0.5(trifluoromethanesulfonate) was dissolved in chloroform (5 mL) and the resulting solution was added to liquid ammonia in chloroform (5 mL) at -70°C. The mixture was allowed to warm to 25 °C over 1 h and then concentrated to dryness in vacuo. The residue was purified by flash chromatography (gradient elution DCM:MeOH 90%-75%) and the resulting solid was redissolved in 10% dichloromethane: methanol (2mL) when Amberlyst resin, mesylate form (400 mg) was added. The mixture was stirred for 2 h then filtered and the filtrate was concentrated in vacuo to give the title compound as a yellow solid (30 mg, 40% yield).
<¾ (DMSO-dg): 9.02-9.05 (1H, d, J=8.6), 8.83-8.85 (1H, d, J=7.8), 8.74-8.75 (1H, d, J=1.2), 8.65 (1H, s), 8.46-8.50 (1H, t, J=8.2), 8.42-8.45 (1H, dd, J=10.0, 2.7), 8.37-8.39 (1H, dd, J=8.4, 1.3), 8.04-8.09 (3H, m), 7.92 (1H, s), 4.72 (3H, s).
m/z (ES+): 345.0 (M+). Example 80 - Preparation of l,ll-difluoro-13-methyl-2-(lH-pyrazol-5-Yl)chromeno[4,3.l2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000159_0001
[00214] 1,1 l-Difluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate (53 mg, 0.086 mmol, Example 18), sodium acetate (15 mg, 0.18 mmol, 2.2 equ), lH-pyrazole-5-boronic acid (14 mg, 0.129 mmol, 1.5 equ), tetrakis triphenylphosphine palladium(O) (10 mg, 10 mol%), 1,2-dimethoxyethane (3.75 mL), and water (1.25 mL) were heated under MW irradiation at 100°C for 15 min. (300W, 2000psi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (gradient elution DCM:MeOH 90%-85%) to give the title compound as an orange solid (7 mg, 15% yield).
<¼ (DMSO-i¾: 13.46 (1H, s), 8.74-8.79 (2H, m), 8.62-8.65 (1H, dd, J=9.9, 2.5), 8.45-8.51 (2H, m), 8.02-8.09 (4H, m), 6.94 (1H, m), 4.70 (3H, d, J=9.7).
m/z (ES+): 386.0 (M+).
Example 81 - Preparation of 4,ll-difluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3.i2- g/¾lphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000159_0002
[00215] A mixture of 4,1 l-difluoro-l 3-methyl-2-(trifluoromethylsulfonyloxy)chromeno [4,3,2- g/z]phenanfhridin-13-ium triflate (100 mg, 0.162 mmol, Example 70), lH-pyrazol-5-ylboronic acid (36 mg, 0.32 mmol, 2 equ), tetrakis(triphenylphosphine)palladium(0) (25 mg, 0.0213 mmol, 0.13 equ) and sodium acetate (38 mg, 0.469 mmol, 2.9 equ) in 2:1 DME:H20 (1.5 mL) was heated under microwave radiation for 15 min. at 100°C (300W, 200psi, run time 30s, cooling system on). The mixture was concentrated to dryness in vacuo and the crude product was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-96%-94%-92%-90%) to give the title compound as an orange solid (25 mg, 29% yield).
& (DMSO-i¾: 13.4 (1H, s), 8.73-8.75 (1H, d, J=8.8), 8.51 (1H, s (br)), 8.45-8.49 (1H, dd, J=8.1), 8.40-8.43 (1H, d, J=9.0), 8.28-8.32 (1H, d, J=13.4), 8.05-8.10 (3H, m), 7.99 (1H, m), 7.22 (1H, s (br)), 4.71 (3H).
m/z (ES+): 386.0 (M+).
Example 82 - Preparation of 4,ll-difluoro-13-methyl-2-(pyrimidin-5-yl)chromeno[4,3^2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000160_0001
[00216] A mixture of 4,11-difluoro-l 3-methyl-2-(trifluoromethylsulfonyloxy)chromeno [4,3,2- g/i]phenanthridin-13-ium triflate (75 mg, 0.121 mmol, Example 70), pyrimidin-5-ylboronic acid (30 mg, 0.240 mmol, 2 equ), tetrakis(triphenylphosphine)palladium(0) (18 mg, 0.016 mmol, 0.13 equ) and sodium acetate (29 mg, 0.352 mmol, 2.9 equ) in 2:1 DME:H20 (1.5 mL) was heated under microwave radiation for 10 min. at 100°C (300W, 200psi, run time 30s, cooling system on). The mixture was concentrated to dryness in vacuo and the crude mixture was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-96%-95%-94%-92%-90%) to give the title compound as a yellow/green solid (40 mg, 61% yield).
4 (DMSO-dg): 9.53 (2H, s), 9.38 (1H, s), 8.79-8.82 (1H, d, J=8.4), 8.58 (1H, s (br)), 8.49-8.54 (1H, dd, J=8.7, 8.0), 8.44-8.49 (1H, d, J=13.6), 8.36-8.40 (1H, dd, J=9.7, 2.4), 8.14-8.16 (1H, d, J=8.3), 8.06-8.11 (2H, m), 4.75 (3H,s ).
m/z (ES+): 398.1 (M+). Example 83 - Preparation of 4Jl-difluoro-2-(2-methoxypyridin-3-yl)-13- methYlchromenor4,3,2-gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000161_0001
[00217] A mixture of 4,1 l-difluoro-l 3-methyl-2-(trifluoromethylsulfonyloxy)chromeno [4,3,2- g/j]phenanthridin-13-ium triflate (75 mg, 0.121 mmol, Example 70), 2-methoxypyridin-3- ylboronic acid (37 mg, 0.240 mmol, 2 equ), tetrakis(triphenylphosphine)palladium(0) (18 mg, 0.016 mmol, 0.13 equ) and sodium acetate (29 mg, 0.352 mmol, 2.9 equ) in 2:1 DME:H20 (1.5 mL) was heated under microwave radiation for 10 min. at 100°C (300W, 200psi, run time 30s, cooling system on). The mixture was concentrated to dryness in vacuo and the crude mixture was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-96%-95%-90%) to give the title compound as a yellow/green solid (50 mg, 71% yield).
<¾ (OMSO-de): 8.77-8.79 (1H, d, J=8.3), 8.48-8.52 (1H, dd, J1=J2=8.2), 8.35-8.41 (3H, m), 8.06-8.18 (5H, m), 7.26-7.29 (1H, dd, J=7.4, 4.9), 4.68 (3H, s), 4.01 (3H, s).
m/z (ES+): 427.1 (M+).
Example 84 - Preparation of ll-fluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Step 1 - Preparation of ll-fluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2- g/i]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000161_0002
[00218] A mixture of l l-fluorochromeno[4,3,2-g/z]phenanthridin-2-yl trifluoromethanesulfonate (700 mg, 1.6 mmol, Example 67), potassium carbonate (330 mg, 2.4 mmol, 1.5 equ) and methyl triflate (1.3 g, 8.0 mmol, 5 equ) was heated at 120°C in a sealed tube for 16 h and was allowed to cool to room temperature. The inorganic materials were removed by filtration and washed with 90% DCM:MeOH. The filtrate was concentrated in vacuo and the residue was purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a yellow solid (600 mg, 62% yield).
& (DMSO-d6): 9.12-9.14 (IH, d, J=9.2), 8.82-8.84 (IH, d, J=8.6), 8.64-8.65 (IH, d, J=2.3), 8.48-8.52 (2H, m), 8.06-8.13 (4H, m), 4.64 (3H, s).
m/z (ES+): 449.9 (M+).
Step 2 - Preparation of ll-fluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2- g/i]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000162_0001
[00219] l l-Fluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g i]phenanthridin- 13-ium trifluoromethanesulfonate (70 mg, 0.11 mmol), sodium acetate (21 mg, 0.25 mmol, 2.2 equ), 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (34 mg, 0.17 mmol, 1.5 equ), tetrakis triphenylphosphine palladium(O) (12 mg, 10 mol%), 1,2-dimethoxyethane (2 mL), and water (0.5 mL) were heated under MW irradiation at 100°C for 20 min. (200W, lOOpsi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (DCM:MeOH 90%) to give the title compound as a red solid (40 mg, 66% yield).
4 (DMSO- 6): 13.29 (IH, s), 8.90-8.93 (IH, d, J= 8.7), 8.75-8.77 (IH, d, J=7.9), 8.70 (IH, s, (br)), 8.46-8.47 (IH, d, J=1.3), 8.38-8.42 (3H, m), 8.23-8.26 (IH, dd, J=8.5, 1.3), 8.00-8.06 (2H, m), 7.94-7.96 (IH, d, J=7.6), 4.72 (3H, s).
m/z (ES+): 368.0 (M+).
Example 85 - Preparation of ll-fluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3<2- gfelphenanthridin- 13-ium trifluoromethanesulfonate
Figure imgf000163_0001
[00220] l l-Fluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/i]phenanthridin- 13-ium trifluoromethanesulfonate (65 mg, 0.10 mmol, Example 84), sodium acetate (20 mg, 0.24 mmol, 2.2 equ), lH-pyrazol-5-ylboronic acid (18 mg, 0.16 mmol, 1.5 equ), tetrakis triphenylphosphine palladium(O) (11 mg, 10 mol%), 1,2-dimethoxyethane (3 mL), and water (1 mL) were heated under MW irradiation at 100°C for 20 min. (200W, lOOpsi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a red solid (37 mg, 66% yield).
<¼ (DMSO-d6): 13.31 (1H, s), 8.96-8.98 (1H, d, J= 8.6), 8.77-8.79 (1H, d, J=8.1), 8.65-8.66 (1H, d, J=1.2), 8.41-8.45 (3H, m), 7.98-8.08 (4H, m), 7.19-7.20 (1H, d, J=2.3), 4.74 (3H, s).
m/z (ES+): 368.0 (M+). Example 86 - Preparation of ll-fluoro-13-methyl-2-(pyrimidin-5-yl)chromeno[4,3^2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000163_0002
[00221] l l-Fluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/i]phenanthridin- 13-ium trifluoromethanesulfonate (63 mg, 0.10 mmol, Example 84), sodium acetate (19 mg, 0.23 mmol, 2.2 equ), pyrimidin-5-ylboronic acid (19 mg, 0.15 mmol, 1.5 equ), tetrakis triphenylphosphine palladium(O) (11 mg, 10 mol%), 1,2-dimethoxyethane (2 mL), and water (1 mL) were heated under MW irradiation at 100°C for 20 min. (200W, lOOpsi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a yellow solid (30 mg, 54% yield). <¾ (DMSO-Je): 9.52 (2H, s), 9.37 (IH, s), 9.09-9.12 (IH, d, J=8.6), 8.87-8.89 (IH, d, J= 7.8), 8.72-8.73 (IH, d, J=1.5), 8.45-8.50 (IH, d, J=8.3), 8.41-8.44 (2H, m), 8.04-8.07 (3H, m), 4.78 (3H, s).
m z (ES+): 380.0 (M+).
Example 87 - Preparation of ll-fluoro-2-(2-methoxypyridin-3-yl)-13- methylchromenor4,3,2-g/tlphenanthridin- -ium trifluoromethanesulfonate
Figure imgf000164_0001
[00222] l l-Fluoro-13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g/i]phenanthridin- 13-ium trifluoromethanesulfonate (90 mg, 0.15 mmol, Example 84), sodium acetate (27 mg, 0.33 mmol, 2.2 equ), 2-methoxypyridin-3-ylboronic acid (34 mg, 0.22 mmol, 1.5 equ), tetrakis triphenylphosphine palladium(O) (15 mg, 10 mol%), 1,2-dimethoxyethane (3 mL), and water (1 mL) were heated under MW irradiation at 100°C for 20 min. (200W, lOOpsi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as an orange solid (47 mg, 56% yield).
<¼ (DMSO-d6): 8.98-9.00 (IH, d, J= 8.6), 8.80-8.82 (IH, d, J=7.8), 8.51-8.52 (IH, d, J=1.3), 8.44-8.48 (IH, t, J=8.2), 8.40-8.42 (IH, dd, J=9.8, 2.5), 8.34-8.36 (IH, dd, J=5.0, 1.9), 8.16-8.19 (IH, dd, J=8.3, 1.4), 8.11-8.14 (IH, dd, J=7.3, 1.9), 8.02-8.06 (3H, m), 7.26-7.29 (IH, dd, J=7.3, 4.9), 4.72 (3H, s), 4.00 (3H, s).
m/z (ES+): 409.0 (M+).
Example 88 - Preparation of 4,ll-difluoro-2-(6-methoxypyridin-2-yl)-13- methylchromenor4,3,2-gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000164_0002
[00223] A mixture of 4,11-difluoro-l 3-methyl-2-(trifluoromethylsulfonyloxy)chromeno [4,3,2- g j]phenanthridin-13-ium triflate (75 mg, 0.121 mmol, Example 70), 2-methoxy-6-(4,4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)pyridine (57 mg, 0.243 mmol, 2 equ), tetrakis(tripnenylphosphine)palladium(0) (14 mg, 0.012 mmol, 0.1 equ) and sodium acetate (22 mg, 0.266 mmol, 2.2 equ) in 2: 1 DME:H20 (1.5 mL) was heated under microwave radiation for 10 min. at 100°C (300W, 200psi, run time 30s, cooling system on). An additional 2 equ of 2- methoxypyridin-3-ylboronic pinacol ester and 0.1 equ of tetrakis(triphenylphosphine)palladium(0) were added and the reaction mixture was heated under microwave radiation for a further 15 min. at 100°C (300W, 200psi, run time 30s, cooling system on). The mixture was concentrated to dryness in vacuo and the crude mixture was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-96%-94%-90%) to give the title compound as a yellow/orange solid (25 mg, 36% yield).
<¼ (OMSO-d6): 8.85 (1H, s), 8.78-8.81 (1H, d, J=8.3), 8.57-8.61 (1H, dd, J=13.9, 1.3), 8.48-8.53 (1H, dd, J=8.3), 8.42-8.45 (1H, dd, J=9.9, 2.4), 8.12-8.14 (1H, dd, J=8.4, 0.6), 8.08-8.10 (2H, m), 8.04-8.06 (1H, dd, J=7.3, 0.6), 7.97-8.01 (1H, dd, J=8.0, 7.6), 7.01-7.03 (1H, dd, J=8.1, 0.5), 4.76 (3H, s), 4.10 (3H, s).
m/z (ES+): 427.0 (M+). Example 89 - Preparation of 4,ll-difluoro-13-methyl-2-(2-oxo-l,2-dihvdropyridin-3- yl)chromeno[4,3,2-gMphenanthridin-13-ium methanesulfonate
Figure imgf000165_0001
[00224] Trimethylsilyl chloride (33 μΕ, 0.26 mmol, 10 equ) was added to a suspension of 4,11- difluoro-2-(2-methoxypyridin-3-yl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium trifluoromethanesulfonate (15 mg, 0.026 mmol, Example 83) and sodium iodide (39 mg, 0.26 mmol, 10 equ) in acetonitrile (500 μί) and the mixture was stirred overnight. The reaction mixture from a separate experiment carried out on 10 mg scale was combined and the mixture was concentrated to dryness in vacuo. The crude product was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution 98%-97%-96%-95%- 94%-92%-90%) to give 14 mg of an orange solid. The product was dissolved in 90% DCM:MeOH and amberlyst A-24 mesylate resin (~150mg) was added. The mixture was stirred for 1 h, then filtered. The filtrate was concentrated in vacuo to give an orange solid. The solid was suspended in water and the resulting suspension was filtered. The solid was dried under suction to give the title compound as an orange solid (2 mg). Dioxane was added to the filtrate and the mixture was left at 4°C overnight. The resulting precipitate was filtered and dried under suction to give the title compound as an orange solid (5 mg, 32% yield)
<¾ (DMSO-d6): 12.2 (1H, s (br)), 8.79 (1H, s), 8.74-8.76 (1H, d, J=8.4), 8.45-9.49 (1H, m), 8.42- 8.44 (1H, dd, J=9.6, 2.4), 8.34-8.38 (1H, dd, J=14.8, 1.2), 8.23-8.25 (1H, dd, J=7.1, 2.0), 8.08- 8.10 (1H ,dd, J=8.4, 0.7), 8.02-8.07 (2H, m), 7.64 (1H, m), 6.48-6.51 (1H, dd, J=7.0), 4.67 (3H, s), 2.29 (3H, s).
m/z (ES+): 413.0 (M+).
Example 90 - Preparation of ll-fluoro-2-(6-methoxypyridin-2-yl)-13- methylchromenor4,3,2-gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000166_0001
[00225] 11-fluoro- 13-methyl-2-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin- 13-ium trifluoromethanesulfonate (75 mg, 0.125 mmol, Example 67), sodium acetate (26 mg, 0.313mmol, 2.5 equ), 6-methoxypyridine-2-boronic acid pinacol ester (44 mg, 0.188 mmol, 1.5 equ), tetrakis triphenylphosphine palladium(O) (14.5 mg, 10 mol%), 1,2-dimethoxyethane (1.125 mL), and water (0.375 mL) were heated under MW irradiation at 100°C for 15 min. (300W, 200psi.). A further amount of 6-methoxypyridine-2-boronic acid pinacol ester (44 mg, 0.188 mmol, 1.5 equ), tetrakis triphenylphosphine palladium(O) (14.5 mg, 10 mol%), was added and then heated under MW irradiation at 100°C for 15 min. (300W, 200psi.). The reaction mixture was then evaporated to dryness, and purified by flash chromatography (DCM:MeOH gradient elution 100%-99%-98%-97%-96%-95%-94%-93%-92%-91%-90%-85%-80%) to give the title compound as a red solid (31 mg, 44% yield). <¾ (DMSO- e): 9.01-9.04 (1H, d, J= 8.7), 8.98-8.99 (1H, d, J=1.4), 8.81-8.83 (1H, d, J=7.8), 8.64-8.67 (1H, dd, J=8.4, 1.4), 8.42-8.48 (2H, m), 7.95-8.06 (5H, m), 6.97-7.00 (1H, dd, J=8.2, 0.8), 4.79 (3H, s), 4.10 (3H, s).
m/z (ES+): 409.0 (M+).
Example 91 - Preparation of ll-fluoro-13-methyl-2-((oxetan-3- yloxY)carbonyl)chromeno[4,3,2-jg¾lphenanthridin-13-ium methanesulfonate
Figure imgf000167_0001
[00226]A suspension of 2-carboxy-l l-fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13- ium 0.5 triflate, 0.5 TFA (150 mg, 0.157 mmol, Example 77) in phosphorus oxychloride (10 mL) was refluxed for 1.5 h. The mixture was cooled to room temperature and concentrated to dryness in vacuo. The residue was co-evaporated several times with toluene, then with diethyl ether to give 2-(chlorocarbonyl)-l l-fluoro-13-methylchromeno[4,3,2-g/?]phenanthridin-13-ium chloride as a dark green gum. The product was used without further purification in the next step.
[00227] A mixture of 3-hydroxyoxetane (100 mL, 1.57 mmol, 10.5 equ) in pyridine (3 mL) was added to approximately 60 mg of crude 2-(chlorocarbonyl)-l l-fluoro-13- methylchromeno[4,3,2-g/i]phenanthridin-13-ium chloride (-0.15 mmol) and the mixture was stirred for 1 h. The mixture was concentrated to dryness in vacuo and the crude product was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCM:MeOH 98%-97%-96%-95%-94%-92%-90%) to give 24 mg of a sticky yellow solid. The product was dissolved in dichloromethane containing a small amount of methanol and the mixture was transferred into a separating funnel. Water was added and the product was extracted with dichloromethane. The combined extracts were dried over magnesium sulphate and concentrated in vacuo to give 15 mg of a yellow solid. The product was dissolved in 90% DCM:MeOH and amberlyst A-24 mesylate resin (~150mg) was added. The mixture was stirred for ~1 h and filtered. The filtrate was concentrated in vacuo and the product was triturated with diethyl ether, filtered and dried under suction to give the title compound as a yellow/orange solid (5.3 mg, 7% yield).
<¼ (DMSO-i¾: 9.07-9.09 (1H, m), 8.86 (1H, m), 8.05-8.50 (3H, m), 8.07-8.19 (3H, m), 5.76 (1H, m), 4.96 (2H, m), 4.78 (2H, m), 4.74 (3H, s), 2.23 (3H, s). m/z (ES+): 401.9 (M+).
Example 92 - Preparation of ll-fluoro-13-methyl-2-(3-methyl-l,2,4-oxadiazoI-5- yl)chromeno[4,3,2-jgMphenanthridin- -ium methanesulfonate
Figure imgf000168_0001
[00228]A mixture of (2-carboxy-ll-fluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium) 0.5(2,2,2-trifluoroacetate) 0.5(trifluoromethanesulfonate) (100 mg, 0.20 mmol; Example 77) in phosphorus oxychloride (5 mL) was heated at 100°C for 1 h and the reaction mixture was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo then co- evaporated with anhydrous toluene to give 2-(chlorocarbonyl)-13-methylchromeno[4,3,2- g/z]phenanthridin-13-ium trifluoromethanesulfonate. Crude 2-(chlorocarbonyl)-13- methylchromeno[4,3,2- i]phenanthridin-13-ium trifluoromethanesulfonate was diluted with anhydrous dichloromethane (10 mL) and triethylamine (30 μί, 0.22 mmol, 1.1 equ) followed by the addition of acetamide oxime (16 mg, 0.125 mmol, 1.1 equ) and the reaction was stirred under an inert atmosphere. After 2 hours, the reaction mixture was diluted with anhydrous toluene, a condenser was fitted and the reaction was heated to 110°C for a further 2 hours. After cooling the solution was concentrated to dryness in vacuo and the residue was purified by flash chromatography (DCM:MeOH 90%) and the resulting solid was redissolved in 10% dichloromethane methanol (2 mL) when Amberlyst resin, mesylate form (200 mg) was added. The mixture was stirred for 2 h then filtered and the filtrate was concentrated in vacuo to give the title compound as a yellow solid (26 mg, 27% yield).
<¾ (DMSO-d6): 9.15-9.17 (1H, d, J=8.6), 8.87-8.89 (2H, m), 8.53-8.56 (1H, dd, J=8.3, 1.4), 8.50- 8.54 (1H, t, J=8.2), 8.40-8.44 (1H, dd, J=9.8, 2.6, 8.13-8.15 (1H, d, J=8.4), 8.05-8.11 (2H, m), 4.77 (3H, s), 2.55 (3H, s), 2.29 (3H, s).
m/z (ES+): 384.0 (M+). Example 93 - Preparation of 2-((l-(tert-butoxycarbonyl)azetidin-3-yloxy)carbonyI)-ll- fluoro-13-methylchromeno[4,3,2-gfelphenanthridin-13-ium methanesulfonate
Figure imgf000169_0001
[00229]A suspension of 2-carboxy-ll-fluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13- ium 0.5 triflate, 0.5 TFA (150 mg, 0.157 mmol, Example 77) in phosphorus oxychloride (10 mL) was refluxed for 1.5 h. The mixture was cooled to room temperature and concentrated to dryness in vacuo. The residue was co-evaporated several times with toluene, then with diethyl ether to give 2-(chlorocarbonyl)-l l-fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium chloride as a dark green gum. The product was used without further purification in the next step.
[00230]A solution of tert-butyl 3-hydroxyazetidine-l-carboxylate (272 mg, 1.57 mmol, 8.4 equ) in pyridine (3 mL) was added to crude 2-(chlorocarbonyl)-l l-fluoro-13-methylchromeno[4,3,2- g/z]phenanthridin-13-ium chloride (75 mg, 0.187 mmol) and the mixture was stirred for 1.5 h, then concentrated to dryness in vacuo. The crude product was dissolved in DCM:MeOH, absorbed on silica and purified by flash chromatography (gradient elution DCMrMeOH 98%- 97%-96%-95%-94%-92%-90%) to give 55 mg of a yellow solid. The product was dissolved in 90% DCMrMeOH and amberlyst A-24 mesylate resin (500 mg) was added. The mixture was stirred for 1 h then filtered. The filtrate was concentrated in vacuo and the product was triturated with diethyl ether, filtered and dried under suction to give 30 mg of a yellow/orange solid. The solid was triturated with ethyl acetate, filtered, washed with diethyl ether and dried under suction to give the title compound as a yellow/orange solid (24 mg, 21% yield).
<¾ (DMSO-i¾): 9.06-9.08 (1H, d, J=8.6), 8.75-8.87 (1H, d, J=8.2), 8.48-8.52 (1H, dd, J=8.0), 8.43-8.46 (1H, dd, J=8.4, 0.7), 8.39-8.42 (1H, dd, J=9.8, 2.4), 8.12-8.14 (1H, d, J=8.3), 8.03-8.09 (2H, m), 5.47-5.52 (1H, m), 4.73 (3H, s), 4.33-4.38 (2H, m), 4.08-4.10 (2H, m), 2.28 (3H, s), 1.42 (9H, s). Example 94 - Preparation of ll-fluoro-13-methyl-2-(2-oxo-l,2-dihvdropyridin-3- yl)chromeno[4,3,2-jg¾lphenanthridin-13-ium chloride
Figure imgf000170_0001
[00231]A flask containing ll-fluoro-2-(6-methoxypyridin-2-yl)-13-methylchromeno[4,3,2- g/i]phenanthridin-13-ium trifluoromethanesulfonate (18 mg, 0.032 mmol, Example 90) in Hydrochloric acid (6M, 5 mL) was fitted with a condenser and stirred at 100°C. After 16 h, the RM was cooled to RT, diluted with water (20 mL) and extracted with dichloromethane (3 x 10 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated in vacuo. The crude material was then subjected to flash chromatography (DCMrMeOH 100%- 99%-98%-97%-96%-95%-90%-85%) to give the title compound as a red solid (3 mg, 21% yield).
m/z (ES+): 394.9 (M+).
Example 95 - Preparation of ll-fluoro-13-methyl-2-(2-oxo-l,2-dihvdropyridin-3- yl)chromeno[4,3,2-g tlphenanthridin-13-ium chloride
Figure imgf000170_0002
[00232]A flask containing ll-fluoro-2-(2-methoxypyridin-3-yl)-13-methylchromeno[4,3,2- g j]phenanthridin-13-ium trifluoromethanesulfonate (70 mg, 0.10 mmol, Example 87) in Hydrochloric acid (6M, 5 mL) was fitted with a condenser and stirred at 100°C. After 16 h, the RM was concentrated to dryness in vacuo. The crude material was then subjected to flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a red solid (15 mg, 28% yield). <¾ (DMSO-J6): 12.16 (IH, s (br)), 8.93-8.95 (IH, d, J=9.0), 8.88-8.89 (IH, d, J=1.2), 8.78-8.80 (IH, d, J=7.8), 8.41-8.46 (2H, m), 8.33-8.36 (IH, dd, J=8.5, 1.4), 8.15-8.17 (IH, dd, J=7.0, 2.0), 7.99-8.05 (2H, m), 7.60 (IH, s (br)), 6.47-6.50 (IH, t, J=6.7), 4.70 (3H, s).
m/z (ES+): 395.0 (M+).
Example 96 - Preparation of 13-methyl-2-(pyrimidin-2-yloxy)chromeno[4,3^2- gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000171_0001
[00233]To a solution of 2-hydroxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium trifluoromethanesulfonate (80 mg, 0.178 mmol, Example 6) in dimethylformamide (2mL) was added sodium hydride 60% dispersion in mineral oil (9 mg, 0.214 mmol, 1.2 equ). After 5 mins, 2-bromopyrimidine (43 mg, 0.267 mmol, 1.5 equ) was added and the mixture was heated to 80°C. After 1 h the reaction was allowed to cool and then concentrated to dryness in vacuo. The product was purified by flash chromatography (gradient elution DCM:MeOH 96%-86%) and the resulting solid was triturated with diethyl ether and filtered to give the title compound as a yellow solid (3 mg, 3% yield).
<¾ (DMSO-c¾: 9.00-9.03 (IH, d, J=9.2), 8.74-8.78 (3H, m), 8.57-8.58 (IH, d, J=7.7), 8.42-8.46 (IH, t, J=8.2), 8.37-8.38 (IH, d, J=2.2), 8.09-8.13 (IH, m), 8.01-8.03 (IH, d, J=9.0), 7.87-7.95 (2H, m), 7.69-7.73 (IH, m), 7.40-7.42 (IH, t, J=4.8), 4.58 (3H, s).
m/z (ES+): 378.0 (M+).
Example 97 - Preparation of ll-fluoro-13-methyl-2-(5-methyl-l,3,4-oxadiazol-2-
Figure imgf000171_0002
[00234]A mixture of (2-carboxy-ll-fluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium) 0.5(2,2,2-trifluoroacetate) 0.5(trifluoromethanesulfonate) (100 mg, 0.20 mmol; Example 77) in dimethylformamide (2.5 mL), diisopropylethylamine (80 mg, 0.6 mmol, 3 equ), O- benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluoro-phosphate (230 mg, 0.6 mmol, 3 equ) and methyl hydrazide (80 mg, 1.1 mmol, 5 equ) was heated under MW irradiation at 50°C for 10 min. (200W, lOOOpsi.). The reaction mixture was then evaporated to dryness, and triturated with ethyl acetate and the residue was purified by flash chromatography (DCM:MeOH 80%) to (2-(2- acetylhydrazinecarbonyl)- 11 -fluoro- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13- ium)0.5(2,2,2-trifluoroacetate)0.5(trifluoromethanesulfonate) (100 mg, 90% yield). This material was then dissolved in phosphorous oxychloride (2 mL) and heated at 100°C for 2 h. The solution was cooled and then concentrated to dryness in vacuo and the residue was purified by flash chromatography (gradient elution DCM:MeOH 90%-85%) and the resulting solid was redissolved in 10% dichloromethane: methanol (2 mL) when silica resin, chloride form (200 mg) was added. The mixture was stirred for 2 h then filtered and the filtrate was concentrated in vacuo to give the title compound as an orange solid (28 mg, 33% yield).
4 (DMSO-i¾): 9.13-9.15 (1H, d, J=8.6), 8.85-8.87 (1H, d, J=7.8), 8.76-8.77 (1H, d, J=1.4), 8.49-8.53 (1H, t, J=7.8), 8.42-8.45 (2H, m), 8.07-8.13 (3H, m), 4.76 (3H, s), 2.71 (3H, s).
m/z (ES+): 384.0 (M+). Example 98 - Preparation of 13-methyl-2-(lH-pyrazol-4-vI)-ll-
(trifluoromethoxy)chromeno[4,3^2-gfelphenanthridin-13-ium trifluoromethanesulfonate Step 1 - Preparation of 13-methyl-ll-(trifluoromethoxy)-2- (trifluoromethylsulfonyloxy)chromeno[4,3,2- i]phenanthridin-13-ium
trifluoromethanesulfonate
Figure imgf000173_0001
A mixture of potassium carbonate (997 mg, 7.07 mmol, 2 equ), 2',3-difluoro-4'- methoxybiphenyl-2-carbonitrile (868 mg, 2.54 mmol, Example 4) and 2-bromo-4- (trifluoromethoxy)phenol (1.0 g, 3.89 mmol, 1.1 equ) in N,N-dimethylformamide (20 mL) was heated at 100°C for 120 hrs and was allowed to cool to room temperature. Saturated ammonium chloride solution (50 mL) was added and the aqueous layer extracted with diethyl ether (3 x 100 mL). The combined organic extracts were dried over magnesium sulphate, filtered and concentrated in vacuo and the residue was purified by flash chromatography (gradient elution Hexanes:EtOAc 80%-50%) to give 3-((2-bromo-4-(trifluoromethoxy)phenoxy)methyl)-2'-fluoro- 4'-methoxybiphenyl-2-carbonitrile as a pale yellow solid (1.255 g, 74% yield), m/z (ES+): 483.9 (MH +(81Br)), 481.9 (MH+(79Br)), which was contaminated with 20% of the 2',3-difluoro-4'- methoxybiphenyl-2-carbonitrile starting material. This material was taken onto the following step without further purification and characterisation in the next step.
7¾ri-butyl lithium (1.6M in pentane, 2.86 mL, 4.58 mmol, 2.1 equ) was added to anhydrous tetrahydrofuran (10 mL) at -78°C and the mixture was stirred for 5 mins. 3-((2-bromo-4- (trifluoromethoxy)phenoxy)methyl)-2'-fluoro-4'-methoxybiphenyl-2-carbonitrile (1.05 g, 2.18 mmol) dissolved in anhydrous tetrahydrofuran (4 mL) was added drop wise over 10 mins and the resulting deep red solution was stirred at -78°C for 10 min when the reaction was allowed to warm to room temperature over 1 h. This was then added to the crude material from a reaction carried out on 20% scale. A saturated solution of ammonium chloride (5 mL) was added and the mixture was then adjusted to pH 14 with 2M sodium hydroxide and then extracted with dichloromethane (3 x 50 mL). The combined organic extracts were dried over magnesium sulphate, filtered and concentrated in vacuo to give 2-methoxy-ll- (trifluoromethoxy)chromeno[4,3,2-g/i]phenanthridine as a red solid (990 mg, assumed 100% conversion), m/z (ES+): 384.0 (MH ). The material was used without further purification and characterization in the next step.
A mixture of 2-methoxy-l l-(trifluoromethoxy)chromeno[4,3,2-g z]phenanthridine (990 mg, 2.59 mmol) and pyridine hydrochloride (2.0 g, 17.3 mmol, 6.7 equ) was heated under microwave radiation for lOmins at 210°C (300W, 150psi, cooling system on, ramp time 30s). Water (20 mL) was added and the resulting precipitate was filtered, washed with water and dried under suction to give ll-(trifluoromethoxy)chromeno[4,3,2-g¾phenanthridin-2-ol (0.840 g, 88% yield over 2 steps), m/z (ES+): 370.0 (MH+). The material was used without further purification and characterization in the next step.
A mixture of ll-(trifluoromethoxy)chromeno[4,3,2-g z]phenanthridin-2-ol (840 mg, 2.27 mmol), N-Phenyl-bis(trifluoromethanesulfonimide) (811 mg, 2.27 mmol, 1 equ) and Ν,Ν- diisopropylethylamine (822 mg, 6.82 mmol, 3 equ) in anhydrous chloroform (20 mL) was heated at 55°C for 16 h and was allowed to cool to room temperature. The mixture was concentrated to dryness in vacuo and the residue was purified by flash chromatography (gradient elution Hexane:CHCi3 40%) to give l l-(trifluoromethoxy)chromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate as a cream solid (590 mg, 40% yield), m/z (ES+): 501.9 (MH+). The material was used without further purification and characterization in the next step.
Methyl triflate (409 mg, 2.5 mmol, 5 equ) was added to a suspension of 11- (trifluoromethoxy)chromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate (250 mg, 0.50 mmol) and potassium carbonate (138 mg, 1.0 mmol, 2 equ) in anhydrous chloroform (5 mL) and the mixture was heated at 110°C for 120 h in a sealed tube. The inorganic materials were removed by filtration and washed with chloroform, concentrated in vacuo and the residue was dissolved in 90% DCM:MeOH. The crude material was filtered and the filtrate as concentrated in vacuo and purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give 13-methyl-ll-(trifluoromethoxy)-2-(trifluoromethylsulfonyloxy)- chromeno[4,3,2-g/z]phenanthridin-13-ium trifluoromethanesulfonate as a yellow solid (130 mg, 39 % yield), and recovered starting material (58 mg).
<¾ (DMSO-i/6): 9.14-9.17 (1H, d, J=9.3), 8.84-8.86 (1H, d, J=8.2), 8.63-8.64 (1H, d, J=2.4), 8.60 (1H, s), 8.50-8.54 (1H, t, J=8.2), 8.19-8.22 (1H, d, J=9.3), 8.10-8.16 (3H, m), 4.65 (3H, s).
m/z (ES+): 515.9 (M+)
Step 2 - Preparation of 13-methyl-2-(lH-pyrazol-4-yl)-ll-
(trifluoromethoxy)chromeno[4,3,2-gA]phenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000175_0001
A mixture of 13-methyl-l l-(trifluoromethoxy)-2-(trifluoromethylsulfonyloxy)-chromeno[4,3,2- gA]phenanthridin-13-ium trifluoromethanesulfonate (60 mg, 0.09 mmol), lH-pyrazole-4-boronic acid pinacol ester (27 mg, 0.135 mmol, 1.5 equ), tetrakis(tripnenylphosphine)palladium(0) (5 mg, 0.004 mmol, 0.04 equ) and potassium acetate (18 mg, 0.18 mmol, 2.0 equ) in DME:H20 3:1 (2 mL) was heated under microwave radiation for 15 min. at 95°C (300W, lOOpsi, cooling system on, ramp time 30s). The mixture was concentrated to dryness in vacuo and purified by flash chromatography (DCM:MeOH 90%) to give the title compound as red/orange solid (42 mg, 80% yield).
<¾ (DMSO- e): 13.3 (1H, s), 8.92-8.94 (1H, d, J=8.7), 8.77-8.79 (1H, d, J=8.2), 8.69 (1H, s), 8.53-8.54 (1H, d, J=2.4), 8.48 (1H, s), 8.40-8.44 (1H, t, J=8.2), 8.32 (1H, s), 8.25-8.27 (1H, dd, J=8.5, 1.2), 8.13-8.16 (1H, dd, J=9.9, 2.2), 8.04-8.06 (1H, d, J=9.3), 7.96-7.98 (1H, d, J=7.9), 4.72 (3H, s).
δρ (DMSO-i¾): -57.2, -77.8
m/z (ES+): 434.0 (M+).
Example 99 - Preparation of 2-(methoxycarbonyl)-13-methyl-ll-
(trifluoromethoxy)chromeno[4,3.i2-gfelphenanthridin-13-ium trifluoromethanesulfonate
Figure imgf000175_0002
1 l-(Trifluoromethoxy)chromeno[4,3,2-g z]phenanthridin-2-yl trifluoromethanesulfonate (290 mg, 0.578 mmol, Example 97), palladium acetate (3 mg, 0.012 mmol, 0.02 equ) and Xantphos (13 mg, 0.023 mmol, 0.04 equ) were put under vacuum in a round bottom flask and the flask was filled with carbon monoxide. A 2: 1 mixture of degassed N,N-dimethylformamide:methanol (5 mL) and triethylamine (242 μί, 1.73 mmol, 3.0 equ) was added and the mixture was heated to 60°C for 1.5 h. After cooling, the crude material was absorbed on silica and the product was purified by flash chromatography (DCM) to yield methyl l l-(trifluoromethoxy)chromeno [4,3,2- g j]phenanthridine-2-carboxylate as an off-white solid (149 mg, 63% yield), m/z (ES+): 412.0 (MH+). The material was used without further characterisation and purification in the next step. Methyl triflate (297 mg, 1.81 mmol, 5 equ) was added to a suspension of 11- (trifluoromethoxy)chromeno[4,3,2- z]phenanthridine-2-carboxylate (149 mg, 0.36 mmol) and potassium carbonate (100 mg, 0.73 mmol, 2 equ) in anhydrous chloroform (4 mL) and the mixture was heated at 110°C for 48 h in a sealed tube, then cooled to room temperature. The inorganic materials were removed by filtration and washed with chloroform, concentrated in vacuo and the residue was dissolved in 90% DCM:MeOH. The crude material was filtered and the filtrate as concentrated in vacuo and purified by flash chromatography (gradient elution DCM:MeOH 95%-90%) to give the title compound as a yellow solid (96 mg, 46% yield).
<¼ (DMSO-d6): 9.07-9.09 (1H, d, J=8.6), 8.86-8.88 (1H, d, J=7.9), 8.76-8.77 (1H, d, J=1.3), 8.50-8.54 (2H, m), 8.40-8.42 (1H, dd, J=8.4, 1.3), 8.18-8.21 (1H, dd, J=9.2, 1.8), 8.09-8.12 (1H, d, J=7.9), 8.03-8.05 (1H, d, J=9.2), 4.73 (3H, s).
Sp (DMSO-i4): -57.2, -77.8
m/z (ES+): 426.0 (M+)

Claims

Figure imgf000177_0001
I
wherein:
X is O, S, SO or S02;
Q is selected from 0\ (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl, (2-6C)alkynyl, or Q is a group of the formula:
-L'-Q1
wherein:
L1 is absent or (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene, each of which is optionally substituted with one or more (l-4C)alkyl groups;
Q1 is selected from -OR9, -NR9R10, -S(0)pR9 (wherein p is 0, 1 or 2), -C(0)R9, - C(0)OR9, -OC(0)R9, -C(O)NR9R10, -N(R10)C(O)R9, -N(Ri0)CON(R10)R9-, -S02N(R9)-, - N(R9)S02-, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
R9 is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
Rio is selected from hydrogen or (l-6C)alkyl; Ri, R2, R3, R4, R5, R6, R7 Rs, R2b R22 and R23 are each independently selected from hydrogen, halogeno, trifluoromethyl, cyano, nitro, hydroxy, mercapto, amino, formyl, carboxy, carbamoyl, ureido, (l-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,
or a group of the formula:
-L2-L3-R20
wherein
L2 is absent or a (l-6C)alkylene, (2-6C)alkenylene or (2-6C)alkynylene;
L3 is absent or is selected from O, S, SO, S02, OS(0)2, N(Ri3), C(O), CH(ORi3), C(0)0, OC(O), C(0)N(R13), N(R13)C(0), N(Ri3)C(0)N(Ri4), S(0)2N(R13), or N(Ri3)S02, wherein Ri3 and RJ4 are each independently selected from hydrogen or (l-4C)alkyl; and
R20 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl-(l-6C)alkyl and wherein R20 is optionally further substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl (l-4C)alkoxy, or a group of the formula:
L4-R24
wherein
L4 is absent or is selected from O, S, SO, S02, N(R25), C(O), CH(OR25),
C(0)0, OC(O), C(0)N(R25), N(R25)C(0), N(R25)C(0)N(R26),
S(0)2N(R25), or N(R25)S02, wherein R¾ and R¾ are each independently selected from hydrogen or (l-4C)alkyl; and
R24 is hydrogen, (l-6C)alkyl, aryl, aryl-(l-6C)alkyl, (3-6C)cycloalkyl, (3- 6C)cycloalkyl-(l-6C)alkyl, (3-6C)cycloalkenyl, (3-6C)cycloalkenyl-(l-
6C)alkyl, heteroaryl, heteroaryl-(l-6C)alkyl, heterocyclyl or heterocyclyl- (l-6C)alkyl and wherein R24 is optionally further substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino, (1-4C) alkyl or (l-4C)alkoxy;
wherein any nitrogen atoms present in any of the groups R1 ; R2, R3, R4, R5, R6, R7 Rg,
R2i, R22 and R23 are optionally in the form of an N-oxide;
or a pharmaceutically acceptable salt or solvate thereof.
2. A compound according to claim 1, wherein X is O, S or S02. A compound according to claim 1 or claim 2, wherein Q is selected from 0~, (l-6C)alkyl, (l-6C)alkoxy, (2-6C)alkenyl or (2-6C)alkynyl, or Q is a group of the formula:
-L'-Q1
wherein:
L1 is (l-6C)alkylene which is optionally substituted with one or more (l-4C)alkyl groups; Q1 is selected from -OR9, -NR9Ri0, -S(0)pR9 (wherein p is 0, 1 or 2), -C(0)R9,
-C(0)OR9, -OC(0)R9, -C(0)NR9Rio, -N(Ri0)C(O)R9, -N(Rio)CON(Ri0)R9-, -S02N(R9)-, -N(R9)S02-, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (3- 8C)cycloalkyl, aryl, heterocyclyl or heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy;
R9 is selected from hydrogen, (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl, and wherein a (l-6C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl, heteroaryl group is optionally substituted by one or more substituents independently selected from halogeno, cyano, nitro, hydroxy, amino or (l-4C)alkoxy; and
Rio is selected from hydrogen or (l-6C)alkyl.
4. A compound according to claim 3, wherein Q is O" or (l-6C)alkyl.
5. A compound according to any one of the preceding claims, wherein Ri, R2, R3, R4, R5, R6, R7 8, R21, R22 and R23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R2o
wherein
L2 is absent or a (l-6C)alkylene or (2-6C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, C(0)N(Ri3),
N(Ri3)C(0), wherein R13 is selected from hydrogen or (l-4C)alkyl; and
R20 is hydrogen, (l-6C)alkyl, heteroaryl, or heterocyclyl and wherein R20 is optionally further substituted by one or more substituents independently selected from halogeno, hydroxy, (l-4C)alkoxy, or a group of the formula: wherein L is N(R25), wherein R25 is independently selected from hydrogen or (1- 4C)alkyl; and
R24 is (l-6C)alkyl;
and wherein any nitrogen atoms present in any of the groups Ri, R2, R3, R4, R5, R6, R7,Rs, R21, R22 and R23 are optionally in the form of a N-oxide.
6. A compound according to claim 5, wherein Rl5 R2, R3, R4, R5, Re, R7 R8, R2i, R22 and R23 are each independently selected from hydrogen, halogeno, hydroxy, or a group of the formula:
-L2-L3-R20
wherein
L2 is absent or a (l-3C)alkylene, or (2-3C)alkynylene;
L3 is absent or is selected from O, OS(0)2, N(Ri3), C(0)0, C(0)N(Ri3),
N(Ri3)C(0), wherein R13 is selected from hydrogen or (l-2C)alkyl; and R20 is hydrogen, (l-3C)alkyl, heteroaryl, or heterocyclyl and wherein R2o is optionally further substituted by one or more substituents independently selected from halogeno, hydroxy, (l-2C)alkoxy;
and wherein any nitrogen atoms present in any of the groups R1 ; R2, R3, R4, R5, R6, R7, Rs, R21, R22 and R23 are optionally in the form of a N-oxide;
7. A compound according to claim 5 or claim 6, wherein one of Ri and R2 is a substituent group other than hydrogen as defined in claim 5 or 6 and the other is hydrogen; and upto four of R3, R4, R5, R6, R7 Rs, R21, R22 and R23 are selected from fluoro, CF3, OCF3 or (1- 2C)alkoxy and the others are hydrogen.
8. A compound according to any one of the preceding claims, wherein
X is O, S or S02;
Q is O" or (l-3C)alkyl;
Ri, R2 and R3 are each as defiend in claim 1, claim 5, claim 6 or claim 7;
R4, R5, R6, R7, Rg, R21, R22 and R23 are each as defined in claim 1, claim 5, claim 6 or claim 7;
or a pharmaceutically acceptable salt or solvate thereof.
9. A compound according to any one of the preceding claims, wherein X is O, S or S02;
Q is O" or methyl;
and Ri, R2, R3, R4, R5, R6, R7, Rs, R21, R22 and R23 each as defiend in claim 7;
or a pharmaceutically acceptable salt or solvate thereof.
10. A compound according to claim 1 , wherein the compound is selected from any one of the following:
13 -methylthiochromeno [4,3 ,2-gh] phenanthridin- 13 -ium;
13-methylchromeno[4,3,2-^/i]phenanthridin-13-ium; or
13-methylthiochromeno[4,3,2-g/z]phenanthridin- 13-ium- 10,10-dioxide;
2-(furan-2-yl)- 13-methylchromeno[4,3,2- /z]phenanthridin- 13-ium;
2-(furan-3-yl)- 13-methylchromeno[4,3,2-g¾phenanthridin- 13-ium;
2-hydroxy-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium;
2-(2-hydroxyethoxy)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
2-acetamido-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium;
13-methyl-2-(N-methylacetamido)chromeno[4,3,2-g z]phenanthridin-13-ium;
2-amino-13-methylthiochromeno[4,3,2-g z]phenanthridin-13-ium;
13-methyl-2-(lH-pyrazol-4-yl)thiochromeno[4,3,2-g i]phenanthridin-13-ium;
2-(3-acetamidoprop- 1 -ynyl)- 13-methylthiochromeno[4,3,2-g¾]phenanthridin- 13-ium;
13-methyl-2-(thiophen-3-yl)thiochromeno[4,3,2-g j]phenanthridin-13-ium;
6, 10-dimethoxy- 13-methylchromeno[4,3 ,2-gh] phenanthridin- 13-ium;
6, 10-dimethoxy- 13-methylchromeno[4,3 ,2-gh] phenanthridin- 13-ium;
6,10-bis(2-hydroxyethoxy)- 13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
1 ,3-difluoro-6, 10-dimethoxy- 13-methylchromeno [4,3 ,2-g i]phenanthridin- 13-ium;
l,l l-difluoro-2-methoxy- 13-methylchromeno[4,3,2-g¾phenanthridin-13-ium;
1,11 -difluoro- 13 -methyl-2-( 1 H-pyrazol-4- yl)chromeno [4,3 ,2-g i]phenanthridin- 13 -ium;
3,9-dimethoxy- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
13-methyl-2-(thiophen-3-yl)chromeno[4,3,2-g z]phenanthridin-13-ium;
2-(3-(dimethylamino)prop-l-ynyl)- 13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
13 -methyl-2-( 1 H-pyrazol-4-yl)chromeno [4,3 ,2-gh] phenanthridin- 13 -ium;
2-methoxy- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
2-(methoxymethyl)-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium;
2-((dimethylamino)methyl)-13-methylchromeno[4,3,2-gA]phenanthridin- 13-ium; 2-methoxy-13-methylthiochromeno[43,2-g z]phenanthridin-13-ium;
2-hydroxy- 13-methylthiochromeno [4,3 ,2-g/z]phenanthridin- 13-ium;
4.11- difluoro-2-methoxy-13-methylchromeno[4,3,2-g¾phenantMdin-13-ium;
2- (3-acetamidoprop- 1 -ynyl)- 13-methylchromeno[4,3,2-g¾]phenanthridin- 13-ium;
2-(dimethylamino)- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
13-methyl-2-morpholinochromeno[4,3,2-g z]phenanthridin-13-ium;
ll-fluoro-3-methoxy-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium;
ll-fluoro-13-methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g z]phenanthridin-13-ium; 13-methyl-2-(pyridin-4-yl)chromeno[4,3,2-g z]phenanthridin-13-ium;
4-(13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium-2-yl)pyridine 1-oxide;
3,11 -difluoro- 13-methylchromeno [4,3,2-g z]phenanthridin- 13-ium;
1.12- difluoro- 13-methylchromeno [4,3,2-g/z]phenanthridin- 13-ium;
1- fluoro-2-methoxy-13-methyl-10-(trifluoromethoxy)chromeno[4,3,2-g z]phenanthridin-13-ium;
3- methoxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
13-methyl-3-(trifluoromethylsulfonyloxy)chromeno[4,3,2-g¾]phenanthridin-13-ium;
13 -methyl-3 -( 1 H-pyrazol-4-yl)chromeno [4,3 ,2-gh] phenanthndin- 13 -ium;
13-methyl-3-(thiophen-3-yl)chromeno[4,3,2-g i]phenanthridin-13-ium;
3-(3-(dimethylamino)prop- 1 -ynyl)- 13-methylchromeno[4,3,2-g i]phenanthridin- 13-ium;
3-(3-acetamidoprop-l-ynyl)-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium;
3,ll-dimethoxy-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
3,11 -difluoro- 13 -methyl-2-( 1 H-pyrazol-4- yl)chromeno [4,3 ,2-g/i]phenanthridin- 13 -ium;
3-fluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium;
3,ll-difluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium;
2- carbamoyl- 1,11 -difluoro- 13-methylchromeno[4,3,2-g i]phenanthridin- 13-ium;
2-carboxy-13-methylchromeno[4,3,2-g¾]phenanthridin-13-ium;
3, 1 l-difluoro-2-methoxy- 13-methylchromeno[4,3 ,2-g/z]phenanthridin- 13-ium;
2-(ethoxycarbonyl)- 13 -methylchromeno [4,3,2-g/z]phenanthridin- 13-ium;
2-(isopropoxycarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
2-carbamoyl- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
2-carbamoyl-3-fluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
4,11 -difluoro-2-(methoxycarbonyl)- 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13-ium;
2-carbamoyl-3,l l-difluoro-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
13-methyl-2-(morpholinomethyl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
1,11 -difluoro-2-(2-hydroxyethoxy) - 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13 -ium; 4-((13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium-2-yl)methyl)morpholine 4-oxide; 2-(2-amino-2-oxoethoxy)- 13-methylchromeno[4,3,2- /i]phenanthridin- 13-ium;
l,l l-difluoro-2-hydroxy-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
3,l l-difluoro-2-(isopropoxycarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium; 2-(methoxymethoxy)-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium;
2-((2-hydroxyethoxy)carbonyl)- 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13-ium;
l l-fluoro-2-(methoxycarbonyl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
2-carbamoyl-4, 11 -difluoro- 13-methylchromeno[4,3,2- z]phenanthridin- 13-ium;
l l-fluoro-2-(methoxymethyl)-13-methylchromeno[4,3,2-g z]phenanthridin-13-ium;
4,1 l-difluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
4, 11 -difluoro- 13-methyl-2-((tetrahydrofuran-2-yloxy)carbonyl)chromeno [4,3 ,2- gA]phenanthridin-13-ium;
4, 1 l-difluoro-2-((3-hydroxypropoxy)carbonyl)- 13-methylchromeno[4,3,2-g¾]phenanthridin- 13- ium;
4,l l-difluoro-2-((2-methoxyethoxy)carbonyl)-13-methylchromeno[4,3,2-g¾phenanthridin-13- ium;
11 -fluoro- 13-methyl-2-(morpholinomethyl)chromeno[4,3,2- z]phenanthridin- 13-ium;
l l-fluoro-13-methyl-2-(3-methyl-l,2,4-oxadiazol-5-yl)chromeno[4,3,2-g/i]phenanthridin-13- ium;
2-((dimethylamino)methyl)- 11 -fluoro- 13-methylchromeno [4,3 ,2-g/z]phenanthridin- 13-ium;
2-carboxy-l l-fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium;
2-(2-amino-2-oxoethoxy)-4, 11 -difluoro- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
2-carbamoyl- 11 -fluoro- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium;
l,l l-difluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
4,l l-difluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3,2-g¾]phenanthridin-13-ium;
4,l l-difluoro-13-methyl-2-(pyrimidin-5-yl)chromeno[4,3,2-g z]phenanthridin-13-ium;
4,l l-difluoro-2-(2-methoxypyridin-3-yl)-13-methylchromeno[43,2-gft]phenanthridin l l-fluoro-13-methyl-2-(lH-pyrazol-4-yl)chromeno[4,3,2-g/i]phenanthridin-13-ium;
l l-fluoro-13-methyl-2-(lH-pyrazol-5-yl)chromeno[4,3,2-g z]phenanthridin-13-ium;
l l-fluoro-13-methyl-2-(pyrimidin-5-yl)chromeno[43,2-g z]phenanthridin-13-ium;
11 -fluoro-2-(2-methoxypyridin-3-yl)- 13-methylchromeno[4,3,2-g¾]phenanthridin- 13-ium;
4,l l-difluoro-2-(6-methoxypyridin-2-yl)-13-methylchromeno[43,2-g z]phenanthridin-13-ium;
4,l l-difluoro-13-methyl-2-(2-oxo-l,2-dihydropyridin-3-yl)chromeno[43,2-g/i]phenanthridin-
13-ium; 11 -fluoro-2-(6-methoxypyridin-2-yl)- 13-methylchromeno[4,3,2-g/z]phenanthridin- 13-ium; ll-fluoro-13-methyl-2-((oxetan-3-yloxy)carbonyl)chromeno[4,3,2- z]phenanthridin-13-ium; ll-fluoro-13-methyl-2-(3-methyl-l,2,4-oxadiazol-5-yl)chromeno[4,3,2-g z]phenanthridin-13- ium;
2-(( 1 -(tert-butoxycarbonyl)azetidin-3-yloxy)carbonyl)- 11 -fluoro- 13-methylchromeno[4,3,2- gA]phenanthridin-13-ium;
11 -fluoro- 13-methyl-2-(2-oxo-l ,2-dihydropyridin-3-yl)chromeno[4,3,2-g z]phenanthridin- 13- ium;
ll-fluoro-13-methyl-2-(2-oxo-l,2-dihydropyridin-3-yl)ch^
ium;
13-methyl-2-(pyrimidin-2-yloxy)chromeno[4,3,2-g z]phenanthridin-13-ium;
ll-fluoro-13-methyl-2-(5-methyl-l,3,4-oxadiazol-2-yl)chromeno[4,3,2-g/i]phenanthridin-13- ium;
Ν,Ν-dimethyl- 1 -(13-methylchromeno[4,3,2- z]phenanthridin- 13-ium-2-yl)methanamine oxide; 2-((2-(dimethylamino)ethoxy)carbonyl)-l l-fluoro-13-memylchromeno[4,3,2-g/i]phenanmridin- 13-ium;
2-( 11 -fluoro- 13-methylchromeno [4,3 ,2-g i]phenanthridin-2— 13-iumcarbonyloxy)-N,N- dimethylethanamine oxide;
2-(3-(dimethylamino)prop- 1 -ynyl)- 11 -fluoro- 13-methylchromeno [4,3 ,2-g z]phenanthridin- 13- ium;
1- (ll-fluoro-13-methylchromeno[4,3,2-g/i]phenanthridin-13-ium-2-yl)-N,N- dimethylmethanamine oxide;
4-((ll-fluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium-2-yl)methyl)morpholine 4- oxide;
ll-fluoro-2-(2-hydroxyethoxy)-13-methylchromeno[4,3,2-g/j]phenanthridin-13-ium;
11 -fluoro-3-(methoxycarbonyl)- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
ll-fluoro-3-(2-hydroxyethoxy)-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium;
2- (3-(dimethylamino)prop-l-ynyl)-l,l l-difluoro-13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium;
2-((dimethylamino)methyl)- 1 , 11 -difluoro- 13 -methylchromeno [4,3 ,2-g/z]phenanthridin- 13-ium; l-(l,ll-difluoro-13-methylchromeno[4,3,2-g/z]phenanthridin-13-ium-2-yl)-N,N- dimethylmethanamine oxide;
1,11 -difluoro- 13-methyl-2-(morpholinomethyl)chromeno [4,3 ,2-g/z]phenanthridin- 13-ium; 4-((l l-difluoro-13-methylchromeno[4,3,2-^/z]phenanthridin-13-ium-2-yl)methyl)m
4-oxide;
2-(ethoxycarbonyl)-3,l l -difluoro-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium;
2-(tert-butoxycarbonyl)-3, 11 -difluoro- 13-methylchromeno[4,3,2-g z]phenanthridin- 13-ium; 3,l l-difluoro-13-methyl-2-((tetrahydrofuran-3-yloxy)carbonyl)chromeno[4,3,2- gA]phenanthridin-13-ium;
6, 10-difluoro-2-(methoxycarbonyl)- 13 -methylchromeno [4,3 ,2-g/i]phenanthridin- 13-ium;
6,10-difluoro-3-(methoxycarbonyl)-13-methylchromeno[4,3,2-g i]phenanthridin-13-ium;
13-methyl-2-(lH-pyrazol-4-yl)- l l-(trifluoromethoxy)chromeno[4,3,2-g¾phenanthridin- 13-ium; 2-(methoxycarbonyl)-13-methyl-l l-(trifluoromethoxy)chromeno[4,3,2-g/i]phenanthridin-13- ium;
in association with a pharmaceutically acceptable anion.
11. A pharmaceutical composition which comprises a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable thereof, in association with a pharmaceutically- acceptable diluent or carrier.
12. A compound according to any one of claims 1 to 10 for use as a medicament. 13. A compound according to any one of claims 1 to 10 for use in the treatment of a proliferative disorder.
14. A compound according to claim 13, wherein the proliferative disorder is cancer. 15. A method of treating a proliferative disorder comprising administering to a human or animal in need of such treatment a therapeutically effective amount of a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt or solvate thereof.
16. A method of inhibiting telomerase activity in a cell, the method comprising administering to said cell compound as claimed in any one of claims 1 to 10, or a pharmaceutically acceptable salt or solvate thereof.
PCT/GB2012/051467 2011-06-24 2012-06-22 Fused pentacyclic anti - proliferative compounds WO2012175991A1 (en)

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GBGB1110727.3A GB201110727D0 (en) 2011-06-24 2011-06-24 Novel anti-proliferative compounds
GBGB1205504.2A GB201205504D0 (en) 2012-03-28 2012-03-28 Novel anti-proliferative compounds
GB1205504.2 2012-03-28

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