WO2017023894A1 - Mthfd2 inhibitors and uses thereof - Google Patents

Mthfd2 inhibitors and uses thereof Download PDF

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WO2017023894A1
WO2017023894A1 PCT/US2016/045086 US2016045086W WO2017023894A1 WO 2017023894 A1 WO2017023894 A1 WO 2017023894A1 US 2016045086 W US2016045086 W US 2016045086W WO 2017023894 A1 WO2017023894 A1 WO 2017023894A1
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indol
methoxy
compound
rel
ring
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PCT/US2016/045086
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French (fr)
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Nello Mainolfi
Mikel MOYER
Eddine Saiah
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Raze Therapeutics, Inc.
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Priority to EP16833713.7A priority Critical patent/EP3331530A4/en
Publication of WO2017023894A1 publication Critical patent/WO2017023894A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
    • C07D209/14—Radicals substituted by nitrogen atoms, not forming part of a nitro radical
    • 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
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to compounds and methods useful for inhibiting methylenetetrahydrofolate dehydrogenase.
  • the invention also provides pharmaceutically acceptable compositions comprising compounds of the present invention and methods of using said compositions in the treatment of various disorders.
  • Mitochondrial glycine (and one-carbon, or 1-C, metabolism) is important in cancer cell metabolism (Locasale, Nat Rev Cancer. 2013 Aug;13(8): 572-83.). Glycine consumption is altered in some cancers, and is a unique predictor of antifolate sensitivity as well as of cancer cell proliferation. Glycine starvation can reduce the proliferation of sensitive cancers.
  • MTHFD2 is a bifunctional enzyme, localized to the mitochondria, that catalyzes two reactions in the mitochondrial 1-C pathway: the dehydrogenase and cyclohydrolase reactions (Tedeschi, et al, (June 22, 2015) Mol Cancer Res, 10.1158/1541-7786.MCR-15- 0117; Christensen and Mackenzie, Vitam Horm. 2008; 79: 393-410.). MTHFD2 is highly upregulated across many cancers relative to normal tissues (Jain, et al., Science. 2012 May 25; 336(6084): 1040-4.).
  • MTHFD2 Genetic silencing of MTHFD2 and inhibition with small molecules slows proliferation across a number of cancer cell lines (Nilsson et al., Nat Commun. 2014; 5: 3128.). Targeting MTHFD2 by small molecule inhibitors could be a therapeutic strategy to reduce cancer cell growth and survival. Accordingly, there remains a need to find MTHFD2 inhibitors useful as therapeutic agents.
  • Compounds of the present invention, and pharmaceutically acceptable compositions thereof, are useful for treating a variety of diseases, disorders or conditions, associated with MTHFD2.
  • diseases, disorders, or conditions include cellular proliferative disorders (e.g., cancer) such as those described herein.
  • Ring A is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenylene, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
  • L 1 is a covalent bond or a bivalent C 1-4 saturated or unsaturated, straight or branched, hydrocarbon chain
  • L 2 is a covalent bond, or a bivalent C 1-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, - N(R)S0 2 -, -S0 2 N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or -S0 2 -;
  • each R is independently hydrogen, or an optionally substituted group selected from Ci-6 aliphatic, phenyl, 4-7 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or:
  • R 1 is hydrogen, optionally substituted Ci-6 aliphatic, or Ring B optionally substituted with 0-4 independently selected R y groups;
  • Ring B is an optionally substituted group selected from phenyl, 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 8-10 membered spirocyclic saturated or partially unsaturated heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur,or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of R x , R y , R 2 and R 3 is independently halogen, R, -OR, -SR, -N(R) 2 , -C(0)R, - C(0)N(R) 2 , -N(R)C(0)R, -N(R)C(0)N(R) 2 , -N(R)C( R)N(R) 2 , -N0 2 , -CN
  • n 0, 1, or 2;
  • n 0, 1, 2, 3, or 4;
  • p is 0 or 1;
  • aliphatic or "aliphatic group”, as used herein, means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as "carbocycle,” “cycloaliphatic” or “cycloalkyl”), that has a single point of attachment to the rest of the molecule.
  • aliphatic groups contain 1-6 aliphatic carbon atoms.
  • aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms.
  • cycloaliphatic (or “carbocycle” or “cycloalkyl”) refers to a monocyclic C3-C6 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule.
  • Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.
  • heteroatom means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR + (as in N-substituted pyrrolidinyl)).
  • unsaturated as used herein, means that a moiety has one or more units of unsaturation.
  • Ci -8 saturated or unsaturated, straight or branched, hydrocarbon chain
  • bivalent Ci -8 or Ci-6) saturated or unsaturated, straight or branched, hydrocarbon chain
  • alkenylene or alkynylene chains that are straight or branched as defined herein.
  • alkylene refers to a bivalent alkyl group.
  • An "alkylene chain” is a polymethylene group, i.e., -(03 ⁇ 4) ⁇ -, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3.
  • a substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.
  • alkenylene refers to a bivalent alkenyl group.
  • a substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.
  • halogen means F, CI, Br, or I.
  • aryl used alone or as part of a larger moiety as in “aralkyl,” “aralkoxy,” or “aryloxyalkyl,” refers to monocyclic or bicyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members.
  • aryl may be used interchangeably with the term “aryl ring.”
  • aryl refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents.
  • aryl is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like.
  • heteroaryl and “heteroar-,” used alone or as part of a larger moiety, e.g., “heteroaralkyl,” or “heteroaralkoxy,” refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 ⁇ electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms.
  • heteroatom refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen.
  • Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl.
  • heteroaryl and “heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring.
  • Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one.
  • heteroaryl group may be mono- or bicyclic.
  • heteroaryl may be used interchangeably with the terms “heteroaryl ring,” “heteroaryl group,” or “heteroaromatic,” any of which terms include rings that are optionally substituted.
  • heteroarylkyl refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.
  • heterocycle As used herein, the terms “heterocycle,” “heterocyclyl,” “heterocyclic radical,” and “heterocyclic ring” are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7-10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above.
  • nitrogen includes a substituted nitrogen.
  • the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), H (as in pyrrolidinyl), or + R (as in N-substituted pyrrolidinyl).
  • a heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted.
  • saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl.
  • heterocycle refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.
  • partially unsaturated refers to a ring moiety that includes at least one double or triple bond.
  • partially unsaturated is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as herein defined.
  • compounds of the invention may contain "optionally substituted” moieties.
  • substituted whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent.
  • an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
  • Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds.
  • stable refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.
  • Suitable monovalent substituents on R° are independently halogen, -(CH 2 )o- 2 R*, -(haloR"), -(CH 2 ) 0 - 2 OH, -(CH 2 ) 0 - 2 OR", -(CH 2 ) 0 - 2 CH(OR") 2 ; -O(haloR'), -CN, -N 3 , -(CH 2 ) 0 - 2 C(O)R e , -(CH 2 ) 0 - 2 C(O)OH, -(CH 2 ) 0 - 2 C(O)OR e , -(CH 2 ) 0 - 2 SR", -(CH 2 )o- 2 SH, -(CH 2 )o- 2 H 2 , -(CH 2 ) 0 - 2 HR*, -(CH 2 )
  • Suitable divalent substituents that are bound to vicinal substitutable carbons of an "optionally substituted” group include: -0(CR * 2 ) 2 - 3 0-, wherein each independent occurrence of R * is selected from hydrogen, Ci- 6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Suitable substituents on the aliphatic group of R * include halogen, -R", -(haloR"), -OH, -OR', -O(haloR'), -CN, -C(0)OH, -C(0)OR', -NH 2 , -NHR", -NR' 2 , or -N0 2 , wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently C 1-4 aliphatic, -CH 2 Ph, -0(CH 2 )o-iPh, or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R ⁇ , - R ⁇ 2 , -C(0)R ⁇ , -C(0)OR ⁇ , -C(0)C(0)R ⁇ , -C(0)CH 2 C(0)R ⁇ , -S(0) 2 R ⁇ , -S(0) 2 R ⁇ 2 , -C(S) R ⁇ 2 , -C( H) R ⁇ 2 , or -N(R ⁇ )S(0) 2 R ⁇ ; wherein each R ⁇ is independently hydrogen, Ci- 6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R ⁇ , taken together with their intervening atom(
  • Suitable substituents on the aliphatic group of R ⁇ are independently halogen, -R", -(haloR*), -OH, -OR', -O(haloR'), -CN, -C(0)OH, -C(0)OR', -NH 2 , - HR", - R' 2 , or
  • each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently aliphatic, -CH 2 Ph, -0(CH 2 )o-iPh, or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • the term "pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference.
  • Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases.
  • Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid
  • organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pect
  • Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (Ci-4alkyl)4 salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.
  • structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention.
  • structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13 C- or 14 C-enriched carbon are within the scope of this invention.
  • Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present invention.
  • an inhibitor is defined as a compound that binds to and /or inhibits MTFIFD2 with measurable affinity.
  • an inhibitor has an IC50 and/or binding constant of less than about 100 ⁇ , less than about 50 ⁇ , less than about 1 ⁇ , less than about 500 nM, less than about 100 nM, less than about 10 nM, or less than about 1 nM.
  • measurable affinity and “measurably inhibit,” as used herein, means a measurable change in MTHFD2 activity between a sample comprising a compound of the present invention, or composition thereof, and MTHFD2, and an equivalent sample comprising MTHFD2, in the absence of said compound, or composition thereof.
  • the present invention provides a compound of formula I:
  • Ring A is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenylene, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
  • L 1 is a covalent bond or a bivalent C 1-4 saturated or unsaturated, straight or branched, hydrocarbon chain;
  • L 2 is a covalent bond, or a bivalent C 1-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, - N(R)S0 2 -, -S0 2 N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or -S0 2 -;
  • each R is independently hydrogen, or an optionally substituted group selected from Ci-6 aliphatic, phenyl, 4-7 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or:
  • R 1 is hydrogen, optionally substituted Ci-6 aliphatic, or Ring B optionally substituted with 0-4 independently selected R y groups;
  • Ring B is an optionally substituted group selected from phenyl, 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 8-10 membered spirocyclic saturated or partially unsaturated heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur,or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of R x , R y , R 2 and R 3 is independently halogen, R, -OR, -SR, -N(R) 2 , -C(0)R, - C(0)N(R) 2 , -N(R)C(0)R, -N(R)C(0)N(R) 2 , -N(R)C( R)N(R) 2 , -N0 2 , -CN
  • n 0, 1, or 2;
  • n 0, 1, 2, 3, or 4;
  • p is 0 or 1;
  • Ring A is bivalent a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenyl ene, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Ring A is phenylene. In other embodiments, Ring A is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring. In some embodiments, Ring A is a 5-6 membered monocyclic saturated or partially unsaturated carbocyclic ring. In some embodiments, Ring A is cyclohexylenyl. In some embodiments, Ring A is cyclopentylenyl. In some embodiments, Ring A is cyclobutylenyl. [0035] In some embodiments, Ring A is a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Ring A a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, Ring A is a 5 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In other embodiments, Ring A is a 6-membered heteroaryl ring having 1-2 nitrogen atoms.
  • Ring A is tetrahydropyranylenyl. In some embodiments, Ring A is piperidinylenyl. In some embodiments, Ring A is pyrrolidinylenyl.
  • Ring A is selected from those depicted in Table 1, below.
  • Ring A is selected from those depicted in Table 2, below.
  • X 1 C(R 2 )-.
  • X 1 N-.
  • X 1 is selected from those depicted in Table 1, below.
  • X 1 is selected from those depicted in Table 2, below.
  • X 2 is selected from those depicted in Table 1, below.
  • X 2 is selected from those depicted in Table 2, below.
  • X 3 is selected from those depicted in Table 1, below.
  • X 3 is selected from those depicted in Table 2, below.
  • ⁇ 1 is - H-.
  • Y 1 is selected from those depicted in Table 1, below.
  • Y 1 is selected from those depicted in Table 2, below.
  • Y 2 is selected from those depicted in Table 1, below.
  • Y 2 is selected from those depicted in Table 2, below.
  • Y 3 is -N-.
  • Y 3 is selected from those depicted in Table 1, below.
  • Y 3 is selected from those depicted in Table 2, below.
  • L 1 is a covalent bond or a bivalent Ci-4 saturated or unsaturated, straight or branched, hydrocarbon chain.
  • L 1 is a covalent bond. In some embodiments, L 1 is a bivalent C 1-4 saturated or unsaturated, straight or branched, hydrocarbon chain.
  • L 1 is methylene
  • L 1 is selected from those depicted in Table 1, below.
  • L 1 is selected from those depicted in Table 2, below.
  • L 2 is a covalent bond, or a bivalent Ci-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, -N(R)S0 2 -, -S0 2 N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or - S0 2 -;
  • L 2 is a covalent bond.
  • L 2 is a bivalent Ci-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, -N(R)S0 2 -, -S0 2 N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or -S0 2 -;
  • L 2 is -N(R)C(0)-. In some embodiments, L 2 is - N(R)C(0) R-. In some embodiments, L 2 is -C(0) R-. [0079] In some embodiments, L 2 is - HC(O)-. In some embodiments, L 2 is - HC(0) H-. In some embodiments, L 2 is -C(0) H-.
  • L 2 is selected from those depicted in Table 1, below.
  • L 2 is selected from those depicted in Table 2, below.
  • R 1 is hydrogen, optionally substituted Ci-6 aliphatic, or Ring B optionally substituted with 0-4 independently selected R y groups.
  • R 1 is hydrogen. In some embodiments, R 1 is optionally substituted Ci- 6 aliphatic. In some embodiments, R 1 is Ring B optionally substituted with 0-4 independently selected R y groups.
  • R 1 is methyl. In some embodiments, R 1 is methoxym ethyl. In some embodiments, R 1 is hydroxyisopropyl.
  • R 1 is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring. In some embodiments, R 1 is a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R 1 is a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • R 1 is cyclopentyl. In some embodiments, R 1 is pyrrolidinyl. In some embodiments, R 1 is tetrahydrofuranyl. In some embodiments, R 1 is tetrahydropyranyl. In some embodiments, R 1 is oxetanyl.
  • R 1 is furanyl. In some embodiments, R 1 is oxazolyl. In some embodiments, R 1 is pyridyl.
  • R 1 is selected from those depicted in Table 1, below.
  • R 1 is selected from those depicted in Table 2, below.
  • each of R x , R y , R 2 and R 3 is independently halogen, R, -OR, - SR, -N(R) 2 , -C(0)R, -C(0)N(R) 2 , -N(R)C(0)R, -N(R)C(0)N(R) 2 , -N(R)C( R)N(R) 2 , -N0 2 , -CN, -S0 2 N(R) 2 , -N(R)S0 2 R, -S0 2 N(R)C(0)R, -S0 2 N(R)C(0)N(R) 2 , S0 2 N(R)C( R)N(R) 2 , -C(0)N(R)C(0)R or -CR ⁇ O ⁇ R.
  • R 2 is R. In some embodiments, R 2 is halogen. In some embodiments, R 2 is -OR.
  • R 2 is methyl. In some embodiments, R 2 is chloro. In some embodiments, R 2 is methoxyl. In some embodiments, R 2 is ethoxyl. [0093] In some embodiments, R 2 is selected from those depicted in Table 1, below.
  • R 2 is selected from those depicted in Table 2, below.
  • R 3 is R.
  • R 3 is methyl
  • R 3 is -CH2OH. In some embodiments, R 3 is - -CH2 H2.
  • R 3 is -C(0)N(R) 2 . In some embodiments, R 3 is -C(0) H 2 .
  • R 3 is selected from those depicted in Table 1, below.
  • R 3 is selected from those depicted in Table 2, below.
  • R x is selected from those depicted in Table 1, below.
  • R x is selected from those depicted in Table 2, below.
  • R y is selected from those depicted in Table 1, below.
  • R y is selected from those depicted in Table 2, below.
  • Ring A, L 1 , L 2 , R 1 , R x , R 2 ,R 3 ,X ⁇ X 2 , X 3 , m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • Ring A, L 1 , L 2 , R 1 , R x , R 2 ,R 3 , m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • Ring A, L 1 , L 2 , R 1 , R x , R 2 ,R 3 , m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • Ring A, L 1 , L 2 , R 1 , R x , R 2 , R 3 , m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • Ring A, L 1 , L 2 , R 1 , R x , R 2 , R 3 , m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • each of R 1 , R x , R 2 , R 3 , R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
  • the present invention provides a compound set forth in Table 1, above, or a pharmaceutically acceptable salt thereof. In some embodiments, the present invention provides a compound set forth in Table 2, above, or a pharmaceutically acceptable salt thereof.
  • compositions are provided.
  • the invention provides a composition comprising a compound of this invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle.
  • the amount of compound in compositions of this invention is such that is effective to measurably inhibit MTHFD2, in a biological sample or in a patient.
  • the amount of compound in compositions of this invention is such that is effective to measurably inhibit MTHFD2, in a biological sample or in a patient.
  • a composition of this invention is formulated for administration to a patient in need of such composition.
  • a composition of this invention is formulated for oral administration to a patient.
  • patient means an animal, preferably a mammal, and most preferably a human.
  • compositions of this invention refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated.
  • Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, di sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxyprop
  • compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir.
  • parenteral as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques.
  • the compositions are administered orally, intraperitoneally or intravenously.
  • Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol.
  • a non-toxic parenterally acceptable diluent or solvent for example as a solution in 1,3-butanediol.
  • acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or di-glycerides.
  • Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions.
  • Other commonly used surfactants such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
  • compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions.
  • carriers commonly used include lactose and corn starch.
  • Lubricating agents such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried cornstarch.
  • aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
  • compositions of this invention may be administered in the form of suppositories for rectal administration.
  • suppositories for rectal administration.
  • suppositories can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug.
  • suitable non-irritating excipient include cocoa butter, beeswax and polyethylene glycols.
  • compositions of this invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
  • Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically- transdermal patches may also be used.
  • compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers.
  • Carriers for topical administration of compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxy ethylene, polyoxypropylene compound, emulsifying wax and water.
  • provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers.
  • Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
  • compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride.
  • the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.
  • compositions of this invention may also be administered by nasal aerosol or inhalation.
  • Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
  • compositions of this invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of this invention are administered with food.
  • compositions of the present invention that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration.
  • provided compositions should be formulated so that a dosage of between 0.01 - 100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.
  • a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated.
  • the amount of a compound of the present invention in the composition will also depend upon the particular compound in the composition.
  • the activity of a compound utilized in this invention as an inhibitor of MTHFD2 may be assayed in vitro, in vivo or in a cell line.
  • In vitro assays include assays that determine inhibition of MTHFD2. Alternate in vitro assays quantitate the ability of the inhibitor to bind to MTHFD2. Detailed conditions for assaying a compound utilized in this invention as an inhibitor of MTHFD2, are set forth in the Examples below.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein.
  • treatment may be administered after one or more symptoms have developed.
  • treatment may be administered in the absence of symptoms.
  • treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.
  • the present invention provides a method for treating a MTHFD2-mediated disorder comprising the step of administering to a patient in need thereof a compound of the present invention, or pharmaceutically acceptable composition thereof.
  • MTHFD2-mediated disorders, diseases, and/or conditions means any disease or other deleterious condition in which MTHFD2, is known to play a role. Accordingly, another embodiment of the present invention relates to treating or lessening the severity of one or more diseases in which MTHFD2 is known to play a role.
  • the methods described herein include methods for the treatment of cancer in a subject.
  • to "treat” means to ameliorate or improve at least one symptom or clinical parameter of the cancer.
  • a treatment can result in a reduction in tumor size or growth rate.
  • a treatment need not cure the cancer or cause remission 100% of the time, in all subjects.
  • the application of agents e.g., inhibitory nucleic acids or small molecules, that inhibit SHMT2 or MTHFD2 reduces cancer cell proliferation and thus treat cancer in subjects.
  • the methods described herein include administering a therapeutically effective dose of one or more agents that inhibit a mitochondrial 1-carbon (1-C) pathway enzyme, e.g., SHMT2, MTHFD2, and/or MTHFD1L.
  • a mitochondrial 1-carbon (1-C) pathway enzyme e.g., SHMT2, MTHFD2, and/or MTHFD1L.
  • cancer refers to cells having the capacity for autonomous growth, i.e., an abnormal state or condition characterized by rapidly proliferating cell growth.
  • the term is meant to include all types of cancerous growths or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues, or organs, irrespective of histopathologic type or stage of invasiveness.
  • tumor refers to cancerous cells, e.g., a mass of cancer cells.
  • Cancers that can be treated or diagnoses using the methods described herein include malignancies of the various organ systems, such as affecting lung, breast, thyroid, lymphoid, gastrointestinal, and genito-urinary tract, as well as adenocarcinomas which include malignancies such as most colon cancers, renal-cell carcinoma, prostate cancer and/or testicular tumors, non-small cell carcinoma of the lung, cancer of the small intestine and cancer of the esophagus.
  • the methods described herein are used for treating or diagnosing a carcinoma in a subject.
  • carcinoma is art recognized and refers to malignancies of epithelial or endocrine tissues including respiratory system carcinomas, gastrointestinal system carcinomas, genitourinary system carcinomas, testicular carcinomas, breast carcinomas, prostatic carcinomas, endocrine system carcinomas, and melanomas.
  • the cancer is renal carcinoma or melanoma.
  • Exemplary carcinomas include those forming from tissue of the cervix, lung, prostate, breast, head and neck, colon and ovary.
  • carcinosarcomas e.g., which include malignant tumors composed of carcinomatous and sarcomatous tissues.
  • An "adenocarcinoma” refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures.
  • sarcoma is art recognized and refers to malignant tumors of mesenchymal derivation.
  • the cancers that are treated by the methods described herein are cancers that have increased levels of glycine uptake or an increased expression or activity of a mitochondrial 1-c enzyme (e.g., SHMT2, MTHFD2, and/or MTHFD1L) relative to normal tissues or to other cancers of the same tissues; methods known in the art and described herein can be used to identify those cancers.
  • a mitochondrial 1-c enzyme e.g., SHMT2, MTHFD2, and/or MTHFD1L
  • the methods include obtaining a sample comprising cells of the cancer, determining the level of glycine uptake or protein, mRNA, or activity of one or more mitochondrial 1-c enzymes (e.g., SHMT2, MTHFD2, and/or MTHFD1L) in the sample, and administering a treatment as described herein (e.g., an antifolate or an agent that inhibits MTHFD2, e.g., ebselen).
  • the cancer is one that is shown herein to have increased levels of glycine uptake.
  • the present invention provides a method for treating one or more disorders, diseases, and/or conditions wherein the disorder, disease, or condition includes, but is not limited to, a cellular proliferative disorder.
  • the present invention features methods and compositions for the diagnosis and prognosis of cellular proliferative disorders (e.g., cancer) and the treatment of these disorders by targeting MTHFD2.
  • cellular proliferative disorders described herein include, e.g., cancer, obesity, and proliferation-dependent diseases. Such disorders may be diagnosed using methods known in the art.
  • Cancers include, without limitation, leukemias (e.g., acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocyte leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain disease, and solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endo
  • proliferative diseases include, e.g., obesity, benign prostatic hyperplasia, psoriasis, abnormal keratinization, lymphoproliferative disorders (e.g., a disorder in which there is abnormal proliferation of cells of the lymphatic system), chronic rheumatoid arthritis, arteriosclerosis, restenosis, and diabetic retinopathy.
  • lymphoproliferative disorders e.g., a disorder in which there is abnormal proliferation of cells of the lymphatic system
  • chronic rheumatoid arthritis e.g., arteriosclerosis, restenosis, and diabetic retinopathy.
  • Proliferative diseases that are hereby incorporated by reference include those described in U.S. Pat. Nos. 5,639,600 and 7,087,648.
  • compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated.
  • the compounds of the invention may be administered orally or parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.
  • Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • the oral compositions can also include adj
  • Injectable preparations for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
  • the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid are used in the preparation of injectables.
  • Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
  • a compound of the present invention In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide.
  • the rate of compound release can be controlled.
  • biodegradable polymers include poly(orthoesters) and poly(anhydrides).
  • Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
  • compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar—agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
  • the active compounds can also be in micro-encapsulated form with one or more excipients as noted above.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art.
  • the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch.
  • Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose.
  • the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
  • buffering agents include polymeric substances and waxes.
  • Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches.
  • the active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required.
  • Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this invention.
  • the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body.
  • Such dosage forms can be made by dissolving or dispensing the compound in the proper medium.
  • Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
  • the invention relates to a method of inhibiting MTHFD2 activity in a biological sample comprising the step of contacting said biological sample with a compound of this invention, or a composition comprising said compound.
  • biological sample includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.
  • Another embodiment of the present invention relates to a method of inhibiting MTHFD2 in a patient comprising the step of administering to said patient a compound of the present invention, or a composition comprising said compound.
  • the invention relates to a method of inhibiting MTHFD2 activity in a patient comprising the step of administering to said patient a compound of the present invention, or a composition comprising said compound.
  • the present invention provides a method for treating a disorder mediated by MTHFD2 in a patient in need thereof, comprising the step of administering to said patient a compound according to the present invention or pharmaceutically acceptable composition thereof. Such disorders are described in detail herein.
  • additional therapeutic agents that are normally administered to treat that condition may also be present in the compositions of this invention.
  • additional therapeutic agents that are normally administered to treat a particular disease, or condition are known as "appropriate for the disease, or condition, being treated.”
  • a compound of the current invention may also be used to advantage in combination with other antiproliferative compounds.
  • antiproliferative compounds include, but are not limited to aromatase inhibitors; anti estrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active compounds; alkylating compounds; histone deacetylase inhibitors; compounds which induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antineoplastic antimetabolites; platin compounds; compounds targeting/decreasing a protein or lipid kinase activity and further anti-angiogenic compounds; compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase; gonadorelin agonists; anti-androgens; methionine aminopeptidase inhibitors; matrix metalloproteinase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of
  • aromatase inhibitor as used herein relates to a compound which inhibits estrogen production, for instance, the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively.
  • the term includes, but is not limited to steroids, especially atamestane, exemestane and formestane and, in particular, non-steroids, especially aminoglutethimide, roglethimide, pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole, fadrozole, anastrozole and letrozole.
  • Exemestane is marketed under the trade name AromasinTM.
  • Formestane is marketed under the trade name LentaronTM.
  • Fadrozole is marketed under the trade name AfemaTM.
  • Anastrozole is marketed under the trade name ArimidexTM.
  • Letrozole is marketed under the trade names FemaraTM or FemarTM.
  • Aminoglutethimide is marketed under the trade name OrimetenTM.
  • a combination of the invention comprising a chemotherapeutic agent which is an aromatase inhibitor is particularly useful for the treatment of hormone receptor positive tumors, such as breast tumors.
  • antiestrogen as used herein relates to a compound which antagonizes the effect of estrogens at the estrogen receptor level.
  • the term includes, but is not limited to tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride.
  • Tamoxifen is marketed under the trade name NolvadexTM.
  • Raloxifene hydrochloride is marketed under the trade name EvistaTM.
  • Fulvestrant can be administered under the trade name FaslodexTM.
  • a combination of the invention comprising a chemotherapeutic agent which is an antiestrogen is particularly useful for the treatment of estrogen receptor positive tumors, such as breast tumors.
  • anti -androgen as used herein relates to any substance which is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (CasodexTM).
  • gonadorelin agonist as used herein includes, but is not limited to abarelix, goserelin and goserelin acetate. Goserelin can be administered under the trade name ZoladexTM.
  • topoisom erase I inhibitor includes, but is not limited to topotecan, gimatecan, irinotecan, camptothecian and its analogues, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148.
  • Irinotecan can be administered, e.g. in the form as it is marketed, e.g. under the trademark CamptosarTM.
  • Topotecan is marketed under the trade name HycamptinTM.
  • topoisom erase II inhibitor includes, but is not limited to the anthracyclines such as doxorubicin (including liposomal formulation, such as CaelyxTM), daunorubicin, epirubicin, idarubicin and nemorubicin, the anthraquinones mitoxantrone and losoxantrone, and the podophillotoxines etoposide and teniposide.
  • Etoposide is marketed under the trade name EtopophosTM.
  • Teniposide is marketed under the trade name VM 26- Bristol Doxorubicin is marketed under the trade name AcriblastinTM or AdriamycinTM.
  • Epirubicin is marketed under the trade name FarmorubicinTM.
  • Idarubicin is marketed, under the trade name ZavedosTM.
  • Mitoxantrone is marketed under the trade name Novantron.
  • microtubule active agent relates to microtubule stabilizing, microtubule destabilizing compounds and microtublin polymerization inhibitors including, but not limited to taxanes, such as paclitaxel and docetaxel; vinca alkaloids, such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate, and vinorelbine; discodermolides; cochicine and epothilones and derivatives thereof.
  • Paclitaxel is marketed under the trade name TaxolTM.
  • Docetaxel is marketed under the trade name TaxotereTM.
  • Vinblastine sulfate is marketed under the trade name Vinblastin R.PTM.
  • Vincristine sulfate is marketed under the trade name FarmistinTM.
  • alkylating agent includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel). Cyclophosphamide is marketed under the trade name CyclostinTM. Ifosfamide is marketed under the trade name HoloxanTM.
  • histone deacetylase inhibitors or "FIDAC inhibitors” relates to compounds which inhibit the histone deacetylase and which possess antiproliferative activity. This includes, but is not limited to, suberoylanilide hydroxamic acid (SAHA).
  • SAHA suberoylanilide hydroxamic acid
  • antimetabolite includes, but is not limited to, 5- fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds, such as 5- azacytidine and decitabine, methotrexate and edatrexate, and folic acid antagonists such as pemetrexed.
  • Capecitabine is marketed under the trade name XelodaTM.
  • Gemcitabine is marketed under the trade name GemzarTM.
  • platinum compound includes, but is not limited to, carboplatin, cis-platin, cisplatinum and oxaliplatin.
  • Carboplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark CarboplatTM.
  • Oxaliplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark EloxatinTM.
  • the term "compounds targeting/decreasing a protein or lipid kinase activity; or a protein or lipid phosphatase activity; or further anti-angiogenic compounds” as used herein includes, but is not limited to, protein tyrosine kinase and/or serine and/or threonine kinase inhibitors or lipid kinase inhibitors, such as a) compounds targeting, decreasing or inhibiting the activity of the platelet-derived growth factor-receptors (PDGFR), such as compounds which target, decrease or inhibit the activity of PDGFR, especially compounds which inhibit the PDGF receptor, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib, SU101, SU6668 and GFB-111; b) compounds targeting, decreasing or inhibiting the activity of the fibroblast growth factor-receptors (FGFR); c) compounds targeting, decreasing or inhibiting the activity of the insulin-like growth factor receptor I (I
  • BCR-Abl kinase and mutants, such as compounds which target decrease or inhibit the activity of c-Abl family members and their gene fusion products, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; or dasatinib (BMS-354825); j) compounds targeting, decreasing or inhibiting the activity of members of the protein kinase C (PKC) and Raf family of serine/threonine kinases, members of the MEK, SRC, JAK/pan-JAK, FAK, PDK1, PKB/Akt, Ras/MAPK, PI3K, SYK, TYK2, BTK and TEC family, and/or members of the cyclin-dependent kinase family (CDK) including staurosporine derivatives, such as midostaurin;
  • c-Met receptor compounds which target, decrease or inhibit the activity of c-Met, especially compounds which inhibit the kinase activity of c-Met receptor, or antibodies that target the extracellular domain of c-Met or bind to HGF
  • PI3K inhibitor includes, but is not limited to compounds having inhibitory activity against one or more enzymes in the phosphatidylinositol-3 -kinase family, including, but not limited to ⁇ , ⁇ , ⁇ , ⁇ , PBK-C2a, PBK-C2p, PBK-C2y, Vps34, pl l0-a, ⁇ 110- ⁇ , ⁇ 110- ⁇ , ⁇ 110- ⁇ , p85-a, ⁇ 85- ⁇ , ⁇ 55- ⁇ , ⁇ 150, plOl, and ⁇ 87.
  • ⁇ inhibitors useful in this invention include but are not limited to ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib.
  • Bcl-2 inhibitor includes, but is not limited to compounds having inhibitory activity against B-cell lymphoma 2 protein (Bcl-2), including but not limited to ABT-199, ABT-731, ABT-737, apogossypol, Ascenta's pan-Bcl-2 inhibitors, curcumin (and analogs thereof), dual Bcl-2/Bcl-xL inhibitors (Infinity Pharmaceuticals/Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and analogs thereof; see WO2008118802), navitoclax (and analogs thereof, see US7390799), NH-1 (Shenayng Pharmaceutical University), obatoclax (and analogs thereof, see WO2004106328), S-001 (Gloria Pharmaceuticals), TW series compounds (Univ. of Michigan), and venetoclax.
  • the Bcl-2 inhibitor is a small molecule therapeutic.
  • the Bcl-2 inhibitor is a small molecule therapeutic.
  • BTK inhibitor includes, but is not limited to compounds having inhibitory activity against Bruton's Tyrosine Kinase (BTK), including, but not limited to AVL-292 and ibrutinib.
  • SYK inhibitor includes, but is not limited to compounds having inhibitory activity against spleen tyrosine kinase (SYK), including but not limited to PRT-062070, R-343, R-333, Excellair, PRT-062607, and fostamatinib
  • PI3K inhibitory compounds and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2004019973, WO2004089925, WO2007016176, US8138347, WO2002088112, WO2007084786, WO2007129161, WO2006122806, WO2005113554, and WO2007044729 the entirety of which are incorporated herein by reference.
  • JAK inhibitory compounds and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2009114512, WO2008109943, WO2007053452, WO2000142246, and WO2007070514, the entirety of which are incorporated herein by reference.
  • Further anti-angiogenic compounds include compounds having another mechanism for their activity, e.g. unrelated to protein or lipid kinase inhibition e.g. thalidomide (ThalomidTM) and TNP-470.
  • proteasome inhibitors useful for use in combination with compounds of the invention include, but are not limited to bortezomib, disulfiram, epigallocatechin-3- gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912, CEP-18770, and MLN9708.
  • Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase are e.g. inhibitors of phosphatase 1, phosphatase 2A, or CDC25, such as okadaic acid or a derivative thereof.
  • Compounds which induce cell differentiation processes include, but are not limited to, retinoic acid, ⁇ - ⁇ - or ⁇ - tocopherol or a- ⁇ - or ⁇ -tocotrienol.
  • cyclooxygenase inhibitor as used herein includes, but is not limited to, Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acid and derivatives, such as celecoxib (CelebrexTM), rofecoxib (VioxxTM), etoricoxib, valdecoxib or a 5-alkyl-2- arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid, lumiracoxib.
  • Cox-2 inhibitors such as celecoxib (CelebrexTM), rofecoxib (VioxxTM), etoricoxib, valdecoxib or a 5-alkyl-2- arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid, lumiracoxib.
  • bisphosphonates includes, but is not limited to, etridonic, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acid.
  • Etridonic acid is marketed under the trade name DidronelTM.
  • Clodronic acid is marketed under the trade name BonefosTM.
  • Tiludronic acid is marketed under the trade name SkelidTM.
  • Pamidronic acid is marketed under the trade name ArediaTM.
  • Alendronic acid is marketed under the trade name FosamaxTM.
  • Ibandronic acid is marketed under the trade name BondranatTM.
  • Risedronic acid is marketed under the trade name ActonelTM.
  • Zoledronic acid is marketed under the trade name ZometaTM.
  • mTOR inhibitors relates to compounds which inhibit the mammalian target of rapamycin (mTOR) and which possess antiproliferative activity such as sirolimus (Rapamune®), everolimus (CerticanTM), CCI-779 and ABT578.
  • heparanase inhibitor refers to compounds which target, decrease or inhibit heparin sulfate degradation.
  • the term includes, but is not limited to, PI-88.
  • biological response modifier refers to a lymphokine or interferons.
  • inhibitor of Ras oncogenic isoforms such as H-Ras, K-Ras, or N-Ras, as used herein refers to compounds which target, decrease or inhibit the oncogenic activity of Ras; for example, a “farnesyl transferase inhibitor” such as L-744832, DK8G557 or Rl 15777 (ZarnestraTM).
  • telomerase inhibitor refers to compounds which target, decrease or inhibit the activity of telomerase. Compounds which target, decrease or inhibit the activity of telomerase are especially compounds which inhibit the telomerase receptor, such as telomestatin.
  • methionine aminopeptidase inhibitor refers to compounds which target, decrease or inhibit the activity of methionine aminopeptidase.
  • Compounds which target, decrease or inhibit the activity of methionine aminopeptidase include, but are not limited to, bengamide or a derivative thereof.
  • proteasome inhibitor refers to compounds which target, decrease or inhibit the activity of the proteasome.
  • Compounds which target, decrease or inhibit the activity of the proteasome include, but are not limited to, Bortezomib (VelcadeTM) and MLN 341.
  • matrix metalloproteinase inhibitor or (“MMP” inhibitor) as used herein includes, but is not limited to, collagen peptidomimetic and nonpeptidomimetic inhibitors, tetracycline derivatives, e.g. hydroxamate peptidomimetic inhibitor batimastat and its orally bioavailable analogue marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551) BMS-279251 , BAY 12-9566, TAA211 , MMI270B or AAJ996.
  • MMP matrix metalloproteinase inhibitor
  • FMS-like tyrosine kinase inhibitors which are compounds targeting, decreasing or inhibiting the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, ⁇ - ⁇ -D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, which are compounds which target, decrease or inhibit anaplastic lymphoma kinase.
  • FMS-like tyrosine kinase receptors are especially compounds, proteins or antibodies which inhibit members of the Flt-3R receptor kinase family, such as PKC412, midostaurin, a staurosporine derivative, SU11248 and MLN518.
  • HSP90 inhibitors includes, but is not limited to, compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90; degrading, targeting, decreasing or inhibiting the HSP90 client proteins via the ubiquitin proteosome pathway.
  • Compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90 are especially compounds, proteins or antibodies which inhibit the ATPase activity of HSP90, such as 17-allylamino, 17-demethoxygeldanamycin (17AAG), a geldanamycin derivative; other geldanamycin related compounds; radicicol and FIDAC inhibitors.
  • antiproliferative antibodies includes, but is not limited to, trastuzumab (HerceptinTM), Trastuzumab-DMl, erbitux, bevacizumab (AvastinTM), rituximab (Rituxan ® ), PR064553 (anti-CD40) and 2C4 Antibody.
  • trastuzumab HerceptinTM
  • Trastuzumab-DMl erbitux
  • bevacizumab AvastinTM
  • rituximab Rasteran ®
  • PR064553 anti-CD40
  • compounds of the current invention can be used in combination with standard leukemia therapies, especially in combination with therapies used for the treatment of AML.
  • compounds of the current invention can be administered in combination with, for example, farnesyl transferase inhibitors and/or other drugs useful for the treatment of AML, such as Daunorubicin, Adriamycin, Ara-C, VP- 16, Teniposide, Mitoxantrone, Idarubicin, Carboplatinum and PKC412.
  • anti-leukemic compounds include, for example, Ara-C, a pyrimidine analog, which is the 2 -alpha-hydroxy ribose (arabinoside) derivative of deoxycytidine. Also included is the purine analog of hypoxanthine, 6-mercaptopurine (6-MP) and fludarabine phosphate.
  • HDAC hi stone deacetylase
  • SAHA suberoylanilide hydroxamic acid
  • HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), Trichostatin A and compounds disclosed in US 6,552,065 including, but not limited to, N-hydroxy-3-[4-[[[2-(2-methyl-lH- indol-3-yl)-ethyl]- amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof and N-hydroxy-3-[4-[(2-hydroxyethyl) ⁇ 2-(lH-indol-3-yl)ethyl]- amino]methyl]phenyl]-2E-2- propenamide, or a pharmaceutically acceptable salt thereof, especially the lactate salt.
  • Somatostatin receptor antagonists as used herein refer to compounds which target, treat or inhibit the somatostatin receptor such as octreotide, and SOM230.
  • Tumor cell damaging approaches refer to approaches such as ionizing radiation.
  • the term "ionizing radiation” referred to above and hereinafter means ionizing radiation that occurs as either electromagnetic rays (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4 th Edition, Vol. 1 , pp. 248-275 (1993).
  • EDG binders and ribonucleotide reductase inhibitors.
  • EDG binders refers to a class of immunosuppressants that modulates lymphocyte recirculation, such as FTY720.
  • ribonucleotide reductase inhibitors refers to pyrimidine or purine nucleoside analogs including, but not limited to, fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6- mercaptopurine (especially in combination with ara-C against ALL) and/or pentostatin.
  • Ribonucleotide reductase inhibitors are especially hydroxyurea or 2-hydroxy-lH-isoindole-l ,3-dione derivatives.
  • VEGF vascular endothelial growth factor
  • l-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof l-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate
  • AngiostatinTM EndostatinTM
  • anthranilic acid amides ZD4190; ZD6474; SU5416; SU6668
  • bevacizumab or anti-VEGF antibodies or anti-VEGF receptor antibodies, such as rhuMAb and RHUFab
  • VEGF aptamer such as Macugon
  • FLT-4 inhibitors, FLT-3 inhibitors VEGFR-2 IgGI antibody
  • Angiozyme RI 4610)
  • Bevacizumab AvastinTM
  • Photodynamic therapy refers to therapy which uses certain chemicals known as photosensitizing compounds to treat or prevent cancers.
  • Examples of photodynamic therapy include treatment with compounds, such as VisudyneTM and porfimer sodium.
  • Angiostatic steroids refers to compounds which block or inhibit angiogenesis, such as, e.g., anecortave, triamcinolone, hydrocortisone, 11-a-epihydrocotisol, cortex ol one, 17a-hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone and dexamethasone.
  • angiogenesis such as, e.g., anecortave, triamcinolone, hydrocortisone, 11-a-epihydrocotisol, cortex ol one, 17a-hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone and dexamethasone.
  • Implants containing corticosteroids refers to compounds, such as fluocinolone and dexamethasone.
  • chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds or compounds with other or unknown mechanism of action.
  • a compound of the current invention may also be used in combination with known therapeutic processes, for example, the administration of hormones or radiation.
  • a provided compound is used as a radiosensitizer, especially for the treatment of tumors which exhibit poor sensitivity to radiotherapy.
  • a compound of the current invention can be administered alone or in combination with one or more other therapeutic compounds, possible combination therapy taking the form of fixed combinations or the administration of a compound of the invention and one or more other therapeutic compounds being staggered or given independently of one another, or the combined administration of fixed combinations and one or more other therapeutic compounds.
  • a compound of the current invention can besides or in addition be administered especially for tumor therapy in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or a combination of these. Long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies, as described above. Other possible treatments are therapy to maintain the patient's status after tumor regression, or even chemopreventive therapy, for example in patients at risk.
  • Those additional agents may be administered separately from an inventive compound-containing composition, as part of a multiple dosage regimen.
  • those agents may be part of a single dosage form, mixed together with a compound of this invention in a single composition. If administered as part of a multiple dosage regime, the two active agents may be submitted simultaneously, sequentially or within a period of time from one another normally within five hours from one another.
  • the term “combination,” “combined,” and related terms refers to the simultaneous or sequential administration of therapeutic agents in accordance with this invention.
  • a compound of the present invention may be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form.
  • the present invention provides a single unit dosage form comprising a compound of the current invention, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.
  • compositions of this invention should be formulated so that a dosage of between 0.01 - 100 mg/kg body weight/day of an inventive compound can be administered.
  • compositions which comprise an additional therapeutic agent that additional therapeutic agent and the compound of this invention may act synergistically. Therefore, the amount of additional therapeutic agent in such compositions will be less than that required in a monotherapy utilizing only that therapeutic agent. In such compositions a dosage of between 0.01 - 1,000 ⁇ g/kg body weight/day of the additional therapeutic agent can be administered.
  • the amount of additional therapeutic agent present in the compositions of this invention will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent.
  • the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent.
  • the compounds of this invention, or pharmaceutical compositions thereof, may also be incorporated into compositions for coating an implantable medical device, such as prostheses, artificial valves, vascular grafts, stents and catheters.
  • an implantable medical device such as prostheses, artificial valves, vascular grafts, stents and catheters.
  • Vascular stents for example, have been used to overcome restenosis (re-narrowing of the vessel wall after injury).
  • patients using stents or other implantable devices risk clot formation or platelet activation. These unwanted effects may be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition comprising a kinase inhibitor.
  • Implantable devices coated with a compound of this invention are another embodiment of the present invention.
  • DIPEA N,N-diisopropylethylamine
  • HATU ⁇ , ⁇ , ⁇ ' , ⁇ ' -tetramethyl-0-(7-azabenzotriazol- 1 -yl)uranium
  • IP A isopropyl alcohol
  • PBS phosphate buffered saline
  • PE petroleum ether
  • TfAA trifluoromethanesulfonic anhydride
  • TIPS triisopropylsilyl
  • Example 2 N-(rel)-(lS,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)furan-2- carboxamide [1-2].
  • Example 3 N-(rel)-(lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrah dr o-2H-pyran-4-carboxamide [1-3] .
  • Example 4 (R)-N-(rel)-((lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-3-carboxamide [1-4].
  • Example 5 N-(rel-(lS,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)oxazole-2- carboxamide [1-5].
  • Example 10 (S)-(rel)-(lS,3R)-3-(5-methoxy-lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine-l-carbox late [I- 10].
  • Example 12 N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-2-hydroxy- 2-methylpropanamide [1-12].
  • Example 13 N-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-2- hydroxyacetamide [1-13].
  • Example 14 and 15 (S)-N-((lR,3S)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-14] and (S)-N-((lS,3R)-3-(5-methoxy- lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-15].
  • Example 16 N-(rel-(lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)acetamide [1-16].
  • Example 17 and 18 (S)-N-((lR,3S)-3-(5-methoxy-l-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-17] and (S)-N-((lS,3R)-3-(5-methoxy- l-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-18].
  • Example 19 and 20 (S)-N-((lR,3S)-3-(5-ethoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-19] and (S)-N-((lS,3R)-3-(5-ethoxy-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-20].
  • Example 21 and 22 (S)-N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-21] and (S)-N-(rel-(lS,3S)-3-(5-chloro- lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-22] .
  • Example 23 and 24 (S)-N-((lS,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-23] and (S)-N-((lR,3S)-3-(5-chloro-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-24].
  • Example 25 and 26 (S)-N-((lS,3S)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)- tetrahydrofuran-2-carboxamide and (S)-N-((lR,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-25 and 1-26].
  • Example 27 (S)-N-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-N- methyltetrahydrofuran-2-carboxamid -27].
  • Example 28 5-chloro-3-(rel-(lS,3R)-3-((S)-tetrahydrofuran-2-carboxamido)- cyclohexyl)-lH-indole-l-carboxami -28].
  • Example 29 rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)-N-(((S)-tetrahydrofuran- 2-yl)methyl)cyclohexanamine [1-29] .
  • Example 30 rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)-N-(((R)-tetrahydrofuran- 2-yl)methyl)cyclohexanamine [1-30] .
  • Example 31 N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydro- 2H-pyran-4-amine [1-31].
  • Example 32 ( ⁇ )-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-3- phenylurea [1-32].
  • Example 33 ( ⁇ )-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-3- methylurea [1-33].
  • Example 34 ( ⁇ )-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-3- (pyridin-2-yl)urea [1-34].
  • Example 35 ( ⁇ )-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)urea [I- 35].
  • Example 36 ( ⁇ )-N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl) pyrrolidine-2-carboxamide [1-36] .
  • Example 37 ( ⁇ )-N-(rel-(lS,3R)-3-(lH-indol-3-yl) cyclohexyl) pyrrolidine-2- carboxamide [1-37].
  • Example 38 and 39 (S)-N-(rel-(lS,3R)-3-(5-chloro-2-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-38] and (S)-N-(rel-(lS,3S)-3-(5-chloro- 2-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-39].
  • Example 40 and 41 (S)-N-(rel-(lS,3R)-3-(5-methoxy-7-methyl-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide [1-40] and (S)-N-(rel-(lS,3S)-3-(5-methoxy- 7-methyl-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide [1-41].
  • Example 42 (S)-N-rel-((lS,3R)-3-(6-(benzyloxy)-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carbox .
  • Example 43 (S)-N-rel-((lS,3R)-3-(6-hydroxy-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide [1-43].
  • Example 44 and 45 (S)-N-((lR,3S)-3-(6-(2-amino-2-oxoethoxy)-5-methoxy- lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide and (S)-N-((l S,3R)-3-(6-(2- amino-2-oxoethoxy)-5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide [1-44] an -45].
  • Example 46 and 47 (S)-N-(rel-(3R,5R)-5-(5-chloro-lH-indol-3-yl) tetrahydro-2H-pyran-3-yl) tetrahydrofuran-2-carboxamide [1-46] and (S)-N-(rel- (3R,5S)-5-(5-chloro-lH-indol-3-yl) tetrahydro-2H-pyran-3-yl) tetrahydrofuran-2- carboxamide [1-47].
  • Example 48 and 49 (S)-N-((lS,3S)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide and (S)-N-(lR,3R)-3-(5-methyl-lH-indol- 3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-48] and [1-49].
  • Example 50 and 51 (S)-N-((lS,3R)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide and (S)-N-((lR,3S)-3-(5-methyl-lH-indol- 3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-50] and [1-51].
  • Example 56 (S)-N-(rel-(lS,3R)-3-(6-(2-hydroxyethyl)-5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-56].
  • the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3 * 30 mL). The combined organic fractions were dried with Na 2 S0 4 , filtered and concentrated. The residue was diluted with DCM (5 mL). To this solution was added Boc 2 0 (87 mg, 0.4 mmol), triethylamine (61 mg, 0.6 mmol) and N,N- Dimethyl-4-aminopyridine (5 mg, 0.04 mmol). The mixture was stirred at room temperature for 3h.
  • Example 57 ( ⁇ )-rel-N-((lS,3S)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)acetamide [1-57].
  • Examples 58 and 59 ((R)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((R)- tetrahydrofuran-2-yl)methanone and ((S)-3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)((R)-tetrahydrofuran-2-yl)methanone [1-58] and [1-59].
  • Example 60 ( ⁇ )-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)methanone [1-60].
  • Example 61 and 62 (S)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)methanone and (R)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)methanone [1-61] and [1-62
  • Example 63 ( ⁇ )-N-cyclopentyl-3-(5-methoxy-lH-indol-3-yl)piperidine-l- carboxamide [1-63].
  • Example 64 and 65 (R)-N-cyclopentyl-3-(5-methoxy-lH-indol-3- yl)piperidine-l-carboxamide and (S)-N-cyclopentyl-3-(5-methoxy-lH-indol-3- yl)piperidine-l-carboxami -64] and [1-65].
  • Example 66 and 67 ((R)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((S)- tetrahydrofuran-2-yl)methanone and ((S)-3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)((S)-tetrahydrofuran-2-yl)methanone [1-66] and [1-67].
  • Example 68 (R)-N-(3-(5-methoxy-lH-indol-3-yl)phenyl)tetrahydrofuran-2- carboxamide [1-68].
  • Example 69 (S)-N-(3-(5-methoxy-lH-indol-3-yl)phenyl)tetrahydrofuran-2- carboxamide [1-69].
  • Human MTHFD2 (residues 36-350 in Uniport ID P13995) was expressed as an N- terminal His 6 tagged protein and purified in E.coli using nickel capture followed by size- exclusion chromatography.
  • MTHFD2 dehydrogenase activity oxidizes methylene tetrahyrofolate to methenyl- tetrahydrofolate with the concomitant reduction of NAD to NADH.
  • MTHFD2 dehydrogenase activity was followed by coupling the generation of NADH to dipahorase catalyzed oxidation of resazurin to fluorescent resorufin.
  • control (DMSO) or compound was incubated with 0.2nM MTHFD2 for 15 minutes in 10.5 ⁇ assay buffer containing lOOmM Hepes, pH 8.0, 5mM MgCb, 5mM KH2PO4, 0.05% BSA, 0.01% Tween- 20 in a 384 well Griener, low volume, black plate.
  • substrate buffer containing 70 ⁇ CH 2 -THF, ImM NAD + , ⁇ Resazurin and 0.42 mg/ml diaphorase was then added to initiate the reaction and the reaction plate was incubated for 30 minutes.
  • Table 3 shows the activity of selected compounds of this invention in the MTHFD2 inhibition assay.
  • the compound numbers correspond to the compound numbers in Tables 1 and 2.
  • Compounds having an activity designated as "A” provided an IC50 ⁇ 10 ⁇ ; compounds having an activity designated as "B” provided an IC50 10 - 30 ⁇ ; compounds having an activity designated as "C” provided an IC50 30 - 50 ⁇ ; compounds having an activity designated as "D” provided an IC50 > 50 ⁇ .

Abstract

The present invention provides compounds, compositions thereof, and methods of using the same.

Description

MTHFD2 INHIBITORS AND USES THEREOF
TECHNICAL FIELD OF THE INVENTION
[0001] The present invention relates to compounds and methods useful for inhibiting methylenetetrahydrofolate dehydrogenase. The invention also provides pharmaceutically acceptable compositions comprising compounds of the present invention and methods of using said compositions in the treatment of various disorders.
BACKGROUND OF THE INVENTION
[0002] Mitochondrial glycine (and one-carbon, or 1-C, metabolism) is important in cancer cell metabolism (Locasale, Nat Rev Cancer. 2013 Aug;13(8): 572-83.). Glycine consumption is altered in some cancers, and is a unique predictor of antifolate sensitivity as well as of cancer cell proliferation. Glycine starvation can reduce the proliferation of sensitive cancers.
[0003] MTHFD2 is a bifunctional enzyme, localized to the mitochondria, that catalyzes two reactions in the mitochondrial 1-C pathway: the dehydrogenase and cyclohydrolase reactions (Tedeschi, et al, (June 22, 2015) Mol Cancer Res, 10.1158/1541-7786.MCR-15- 0117; Christensen and Mackenzie, Vitam Horm. 2008; 79: 393-410.). MTHFD2 is highly upregulated across many cancers relative to normal tissues (Jain, et al., Science. 2012 May 25; 336(6084): 1040-4.). Genetic silencing of MTHFD2 and inhibition with small molecules slows proliferation across a number of cancer cell lines (Nilsson et al., Nat Commun. 2014; 5: 3128.). Targeting MTHFD2 by small molecule inhibitors could be a therapeutic strategy to reduce cancer cell growth and survival. Accordingly, there remains a need to find MTHFD2 inhibitors useful as therapeutic agents.
SUMMARY OF THE INVENTION
[0004] It has now been found that compounds of this invention, and pharmaceutically acceptable compositions thereof, are effective as MTHFD2 inhibitors. Such compounds have the general formula I:
Figure imgf000003_0001
or a pharmaceutically acceptable salt thereof, wherein each variable is as defined and described herein.
[0005] Compounds of the present invention, and pharmaceutically acceptable compositions thereof, are useful for treating a variety of diseases, disorders or conditions, associated with MTHFD2. Such diseases, disorders, or conditions include cellular proliferative disorders (e.g., cancer) such as those described herein.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS
1. General Description of Certain Embodiments of the Invention:
[0006] Compounds of the present invention, and compositions thereof, are useful as inhibitors of MTHFD2. In certain embodiments, the present invention provides a compound of formula I:
Figure imgf000003_0002
I
or a pharmaceutically acceptable salt thereof, wherein:
Ring A is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenylene, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
L1 is a covalent bond or a bivalent C1-4 saturated or unsaturated, straight or branched, hydrocarbon chain; L2 is a covalent bond, or a bivalent C1-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, - N(R)S02-, -S02N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or -S02-;
each R is independently hydrogen, or an optionally substituted group selected from Ci-6 aliphatic, phenyl, 4-7 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or:
two R groups on the same nitrogen are taken together with their intervening atoms to form a 4-7 membered monocyclic saturated, partially unsaturated, or heteroaryl ring having 0-3 heteroatoms, in addition to the nitrogen, independently selected from nitrogen, oxygen, or sulfur;
R1 is hydrogen, optionally substituted Ci-6 aliphatic, or Ring B optionally substituted with 0-4 independently selected Ry groups;
Ring B is an optionally substituted group selected from phenyl, 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 8-10 membered spirocyclic saturated or partially unsaturated heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur,or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of Rx, Ry, R2 and R3 is independently halogen, R, -OR, -SR, -N(R)2, -C(0)R, - C(0)N(R)2, -N(R)C(0)R, -N(R)C(0)N(R)2, -N(R)C( R)N(R)2, -N02, -CN, - S02N(R)2, -N(R)S02R, -S02N(R)C(0)R, -S02N(R)C(0)N(R)2, S02N(R)C( R)N(R)2 , -C(0)N(R)C(0)R or -CR^O^R; and
m is 0, 1, or 2;
n is 0, 1, 2, 3, or 4;
p is 0 or 1;
X1 is =C(R2)- or =N-;
each of X2 and X3 is -C(R2)= or -N=;
each == is a double bond or a single bond, as valency allows;
Y1 is -N(R3)- or -N=;
Y2 is -C(R3)=, =C(R3)-, =N-, or -N=; and Y3 is -C= or -N-
2. Compounds and Definitions:
[0007] Compounds of the present invention include those described generally herein, and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Additionally, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th Ed., Ed.: Smith, M B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.
[0008] The term "aliphatic" or "aliphatic group", as used herein, means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as "carbocycle," "cycloaliphatic" or "cycloalkyl"), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C6 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.
[0009] The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR+ (as in N-substituted pyrrolidinyl)). [0010] The term "unsaturated," as used herein, means that a moiety has one or more units of unsaturation.
[0011] As used herein, the term "bivalent Ci-8 (or Ci-6) saturated or unsaturated, straight or branched, hydrocarbon chain", refers to bivalent alkylene, alkenylene, and alkynylene chains that are straight or branched as defined herein.
[0012] The term "alkylene" refers to a bivalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -(0¾)η-, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.
[0013] The term "alkenylene" refers to a bivalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.
[0014] The term "halogen" means F, CI, Br, or I.
[0015] The term "aryl" used alone or as part of a larger moiety as in "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to monocyclic or bicyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments of the present invention, "aryl" refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term "aryl," as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like.
[0016] The terms "heteroaryl" and "heteroar-," used alone or as part of a larger moiety, e.g., "heteroaralkyl," or "heteroaralkoxy," refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 π electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms "heteroaryl" and "heteroar-", as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one. A heteroaryl group may be mono- or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which terms include rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.
[0017] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocyclic ring" are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7-10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), H (as in pyrrolidinyl), or + R (as in N-substituted pyrrolidinyl).
[0018] A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical," are used interchangeably herein, and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. A heterocyclyl group may be mono- or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.
[0019] As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as herein defined.
[0020] As described herein, compounds of the invention may contain "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally" or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.
[0021] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen; -(CH2)o-4R°; -(CH2)0^OR°; -0(CH2)o-4R°, - 0-(CH2)o-4C(0)OR°; -(CH2)0^CH(OR°)2; -(CH2)0-4SR°; -(CH2)0^Ph, which may be substituted with R°;
Figure imgf000008_0001
which may be substituted with R°; -CH=CHPh, which may be substituted with R°; -(CH2)o^O(CH2)o-i-pyridyl which may be substituted with R°; -N02; -CN; -N3; -(CH2)0^N(R°)2; -(CH2)0-4N(R°)C(O)R°; -N(R°)C(S)R°; - (CH2)o-4N(R°)C(0) R°2; -N(R°)C(S) R°2; -(CH2)0-4N(R°)C(O)OR°; -N(R°)N(R°)C(0)R°; -N(R°)N(R°)C(0) R°2; -N(R°)N(R°)C(0)OR°; -(CH2)0^C(O)R°; -C(S)R°; -(CH2)0- 4C(0)OR°; -(CH2)o-4C(0)SR°; -(CH2)0^C(O)OSiR°3; -(CH2)0-4OC(O)R°; -OC(O)(CH2)0- 4SR- SC(S)SR°; -(CH2)o-4SC(0)R°; -(CH2)0^C(O) R°2; -C(S) R°2; -C(S)SR°; - SC(S)SR°, -(CH2)o-4OC(0) R°2; -C(0)N(OR°)R°; -C(0)C(0)R°; -C(0)CH2C(0)R°; - C(NOR°)R°; -(CH2)o^SSR°; -(CH2)0^S(O)2R°; -(CH2)0-4S(O)2OR°; -(CH2)0^OS(O)2R°; - S(0)2 R°2; -(CH2)o^S(0)R°; -N(R°)S(0)2 R°2; -N(R°)S(0)2R°; -N(OR°)R°; - C( H) R°2; -P(0)2R°; -P(0)R°2; -OP(0)R°2; -OP(0)(OR°)2; SiR°3; -(Ci_4 straight or branched alkylene)0-N(R°)2; or -(Ci-4 straight or branched alkylene)C(0)0-N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, Ci-6 aliphatic, -CH2Ph, -0(CH2)o-iPh, -CH2-(5-6 membered heteroaryl ring), or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.
[0022] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), are independently halogen, -(CH2)o-2R*, -(haloR"), -(CH2)0-2OH, -(CH2)0-2OR", -(CH2)0-2CH(OR")2; -O(haloR'), -CN, -N3, -(CH2)0-2C(O)Re, -(CH2)0-2C(O)OH, -(CH2)0-2C(O)ORe, -(CH2)0- 2SR", -(CH2)o-2SH, -(CH2)o-2 H2, -(CH2)0-2 HR*, -(CH2)0-2 R"2, -N02, -SiR"3, -OSiR"3, -C(0)SR' -(Ci-4 straight or branched alkylene)C(0)OR', or -SSR' wherein each R' is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently selected from Ci-4 aliphatic, -CH2Ph, -0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S.
[0023] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =0, =S, =N R* 2, =N HC(0)R*, = HC(0)OR*, =N HS(0)2R*, = R*, =NOR*, -0(C(R* 2))2-30- or -S(C(R* 2))2_3S- wherein each independent occurrence of R* is selected from hydrogen, Ci-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an "optionally substituted" group include: -0(CR* 2)2-30-, wherein each independent occurrence of R* is selected from hydrogen, Ci-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0024] Suitable substituents on the aliphatic group of R* include halogen, -R", -(haloR"), -OH, -OR', -O(haloR'), -CN, -C(0)OH, -C(0)OR', -NH2, -NHR", -NR'2, or -N02, wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently C 1-4 aliphatic, -CH2Ph, -0(CH2)o-iPh, or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0025] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R†, - R† 2, -C(0)R†, -C(0)OR†, -C(0)C(0)R†, -C(0)CH2C(0)R†, -S(0)2R†, -S(0)2 R† 2, -C(S) R† 2, -C( H) R† 2, or -N(R†)S(0)2R†; wherein each R† is independently hydrogen, Ci-6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R†, taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0026] Suitable substituents on the aliphatic group of R† are independently halogen, -R", -(haloR*), -OH, -OR', -O(haloR'), -CN, -C(0)OH, -C(0)OR', -NH2, - HR", - R'2, or
-N02, wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently
Figure imgf000010_0001
aliphatic, -CH2Ph, -0(CH2)o-iPh, or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0027] As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like.
[0028] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(Ci-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.
[0029] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13C- or 14C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present invention.
[0030] As used herein, the term "inhibitor" is defined as a compound that binds to and /or inhibits MTFIFD2 with measurable affinity. In certain embodiments, an inhibitor has an IC50 and/or binding constant of less than about 100 μΜ, less than about 50 μΜ, less than about 1 μΜ, less than about 500 nM, less than about 100 nM, less than about 10 nM, or less than about 1 nM. [0031] The terms "measurable affinity" and "measurably inhibit," as used herein, means a measurable change in MTHFD2 activity between a sample comprising a compound of the present invention, or composition thereof, and MTHFD2, and an equivalent sample comprising MTHFD2, in the absence of said compound, or composition thereof.
3. Description of Exemplary Embodiments:
[0032] In certain embodiments, the present invention provides a compound of formula I:
Figure imgf000012_0001
I
or a pharmaceutically acceptable salt thereof, wherein:
Ring A is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenylene, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
L1 is a covalent bond or a bivalent C1-4 saturated or unsaturated, straight or branched, hydrocarbon chain;
L2 is a covalent bond, or a bivalent C1-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, - N(R)S02-, -S02N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or -S02-;
each R is independently hydrogen, or an optionally substituted group selected from Ci-6 aliphatic, phenyl, 4-7 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or:
two R groups on the same nitrogen are taken together with their intervening atoms to form a 4-7 membered monocyclic saturated, partially unsaturated, or heteroaryl ring having 0-3 heteroatoms, in addition to the nitrogen, independently selected from nitrogen, oxygen, or sulfur;
R1 is hydrogen, optionally substituted Ci-6 aliphatic, or Ring B optionally substituted with 0-4 independently selected Ry groups;
Ring B is an optionally substituted group selected from phenyl, 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 8-10 membered spirocyclic saturated or partially unsaturated heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur,or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of Rx, Ry, R2 and R3 is independently halogen, R, -OR, -SR, -N(R)2, -C(0)R, - C(0)N(R)2, -N(R)C(0)R, -N(R)C(0)N(R)2, -N(R)C( R)N(R)2, -N02, -CN, - S02N(R)2, -N(R)S02R, -S02N(R)C(0)R, -S02N(R)C(0)N(R)2, S02N(R)C( R)N(R)2 , -C(0)N(R)C(0)R or -CR^O^R; and
m is 0, 1, or 2;
n is 0, 1, 2, 3, or 4;
p is 0 or 1;
X1 is =C(R2)- or =N-;
each of X1 and X2 is -C(R2)= or -N=;
each == is double or single bond as valency allows;
Y1 is -N(R3)- or -N=;
Y2 is -C(R3)=, =C(R3)-, =N-, or -N=; and
Y3 is -C= or -N-.
[0033] As defined generally above, Ring A is bivalent a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenyl ene, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0034] In some embodiments, Ring A is phenylene. In other embodiments, Ring A is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring. In some embodiments, Ring A is a 5-6 membered monocyclic saturated or partially unsaturated carbocyclic ring. In some embodiments, Ring A is cyclohexylenyl. In some embodiments, Ring A is cyclopentylenyl. In some embodiments, Ring A is cyclobutylenyl. [0035] In some embodiments, Ring A is a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, Ring A a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, Ring A is a 5 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In other embodiments, Ring A is a 6-membered heteroaryl ring having 1-2 nitrogen atoms.
[0036] In some embodiments, Ring A is tetrahydropyranylenyl. In some embodiments, Ring A is piperidinylenyl. In some embodiments, Ring A is pyrrolidinylenyl.
[0037] In some embodiments, Ring A is selected from those depicted in Table 1, below.
[0038] In some embodiments, Ring A is selected from those depicted in Table 2, below.
[0039] As defined generally above, X1 is =C(R2)- or =N-.
[0040] In some embodiments, X1 is =C(R2)-.
[0041] In some embodiments, X1 is =N-.
[0042] In some embodiments, X1 is selected from those depicted in Table 1, below.
[0043] In some embodiments, X1 is selected from those depicted in Table 2, below.
[0044] As defined generally above, X2 is -C(R2)= or -N=.
[0045] In some embodiments, X2 is -C(R2)=.
[0046] In some embodiments, X2 is -N=.
[0047] In some embodiments, X2 is selected from those depicted in Table 1, below.
[0048] In some embodiments, X2 is selected from those depicted in Table 2, below.
[0049] As defined generally above, X3 is -C(R2)= or -N=.
[0050] In some embodiments, X3 is -C(R2)=.
[0051] In some embodiments, X3 is -N=.
[0052] In some embodiments, X3 is selected from those depicted in Table 1, below.
[0053] In some embodiments, X3 is selected from those depicted in Table 2, below.
[0054] As defined generally above, Y1 is -N(R3)- or -N=.
[0055] In some embodiments^1 is - H-.
[0056] In some embodiments^1 is -N=.
[0057] In some embodiments, Y1 is selected from those depicted in Table 1, below.
[0058] In some embodiments, Y1 is selected from those depicted in Table 2, below. [0059] As generally defined above, Y2 is -C(R3)=, =C(R3)-, =N-, or -N=;
[0060] In some embodiments, Y2 is -C(R3)=.
[0061] In some embodiments, Y2 is =C(R3)-.
[0062] In some embodiments, Y2 is =N-.
[0063] In some embodiments, Y2 is -N=.
[0064] In some embodiments, Y2 is selected from those depicted in Table 1, below.
[0065] In some embodiments, Y2 is selected from those depicted in Table 2, below.
[0066] As generally defined above, Y3 is -C= or -N-.
[0067] In some embodiments, Y3 is -C=.
[0068] In some embodiments, Y3 is -N-.
[0069] In some embodiments, Y3 is selected from those depicted in Table 1, below.
[0070] In some embodiments, Y3 is selected from those depicted in Table 2, below.
[0071] As defined generally above, L1 is a covalent bond or a bivalent Ci-4 saturated or unsaturated, straight or branched, hydrocarbon chain.
[0072] In some embodiments, L1 is a covalent bond. In some embodiments, L1 is a bivalent C 1-4 saturated or unsaturated, straight or branched, hydrocarbon chain.
[0073] In some embodiments, L1 is methylene.
[0074] In some embodiments, L1 is selected from those depicted in Table 1, below.
[0075] In some embodiments, L1 is selected from those depicted in Table 2, below.
[0076] As defined generally above, L2 is a covalent bond, or a bivalent Ci-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, -N(R)S02-, -S02N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or - S02-;
[0077] In some embodiments, L2 is a covalent bond. In some embodiments, L2 is a bivalent Ci-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, -N(R)S02-, -S02N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or -S02-;
[0078] In some embodiments, L2 is -N(R)C(0)-. In some embodiments, L2 is - N(R)C(0) R-. In some embodiments, L2 is -C(0) R-. [0079] In some embodiments, L2 is - HC(O)-. In some embodiments, L2 is - HC(0) H-. In some embodiments, L2 is -C(0) H-.
[0080] In some embodiments, L2 is selected from those depicted in Table 1, below.
[0081] In some embodiments, L2 is selected from those depicted in Table 2, below.
[0082] As defined generally above, R1 is hydrogen, optionally substituted Ci-6 aliphatic, or Ring B optionally substituted with 0-4 independently selected Ry groups.
[0083] In some embodiments, R1 is hydrogen. In some embodiments, R1 is optionally substituted Ci-6 aliphatic. In some embodiments, R1 is Ring B optionally substituted with 0-4 independently selected Ry groups.
[0084] In some embodiments, R1 is methyl. In some embodiments, R1 is methoxym ethyl. In some embodiments, R1 is hydroxyisopropyl.
[0085] In some embodiments, R1 is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring. In some embodiments, R1 is a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R1 is a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0086] In some embodiments, R1 is cyclopentyl. In some embodiments, R1 is pyrrolidinyl. In some embodiments, R1 is tetrahydrofuranyl. In some embodiments, R1 is tetrahydropyranyl. In some embodiments, R1 is oxetanyl.
[0087] In some embodiments, R1 is furanyl. In some embodiments, R1 is oxazolyl. In some embodiments, R1 is pyridyl.
[0088] In some embodiments, R1 is selected from those depicted in Table 1, below.
[0089] In some embodiments, R1 is selected from those depicted in Table 2, below.
[0090] As defined above, each of Rx, Ry, R2 and R3 is independently halogen, R, -OR, - SR, -N(R)2, -C(0)R, -C(0)N(R)2, -N(R)C(0)R, -N(R)C(0)N(R)2, -N(R)C( R)N(R)2, -N02, -CN, -S02N(R)2, -N(R)S02R, -S02N(R)C(0)R, -S02N(R)C(0)N(R)2, S02N(R)C( R)N(R)2 , -C(0)N(R)C(0)R or -CR^O^R.
[0091] In some embodiments, R2 is R. In some embodiments, R2 is halogen. In some embodiments, R2 is -OR.
[0092] In some embodiments, R2 is methyl. In some embodiments, R2 is chloro. In some embodiments, R2 is methoxyl. In some embodiments, R2 is ethoxyl. [0093] In some embodiments, R2 is selected from those depicted in Table 1, below.
[0094] In some embodiments, R2 is selected from those depicted in Table 2, below.
[0095] In some embodiments, R3 is R.
[0096] In some embodiments, R3 is methyl.
[0097] In some embodiments, R3 is -CH2OH. In some embodiments, R3 is - -CH2 H2.
[0098] In some embodiments, R3 is -C(0)N(R)2. In some embodiments, R3 is -C(0) H2.
[0099] In some embodiments, R3 is selected from those depicted in Table 1, below.
[00100] In some embodiments, R3 is selected from those depicted in Table 2, below.
[00101] In some embodiments, Rx is selected from those depicted in Table 1, below.
[00102] In some embodiments, Rx is selected from those depicted in Table 2, below.
[00103] In some embodiments, Ry is selected from those depicted in Table 1, below.
[00104] In some embodiments, Ry is selected from those depicted in Table 2, below.
[00105] In certain embodiments, the present invention provides a compound of formula I, wherein Y1 is - H-, Y2 is -C(R3)=, and Y3 is -C=, thereby forming a compound of formula
I-A:
Figure imgf000017_0001
I-A
or a pharmaceutically acceptable salt thereof, wherein each of Ring A, L1, L2, R1, Rx, R2,R3,X\ X2, X3, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00106] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -N= X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)= and Y3 is -C=, thereby forming a compound of formula I-A-l:
Figure imgf000018_0001
I-A-l
or a pharmaceutically acceptable salt thereof, wherein each of Ring A, L1, L2, R1, Rx, R2,R3, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00107] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -N= Y1 is - H-, Y2 -C(R3)= and Y3 is -C=, thereby forming a compound of formula I-A-2:
Figure imgf000018_0002
I-A-2
or a pharmaceutically acceptable salt thereof, wherein each of Ring A, L1, L2, R1, Rx, R2,R3, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00108] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)= and Y3 is -C=, thereby forming a compound of formula I-A-3:
Figure imgf000018_0003
I-A-3
or a pharmaceutically acceptable salt thereof, wherein each of Ring A, L1, L2, R1, Rx, R2, R3, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00109] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =Ν-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)= and Y3 is -C=, thereby forming a compound of formula I-A-4:
Figure imgf000019_0001
I-A-4
or a pharmaceutically acceptable salt thereof, wherein each of Ring A, L1, L2, R1, Rx, R2, R3, m, n and p is as defined above and described in embodiments herein, both singly and in combination
[00110] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is - H-, Y2 -N= and Y3 is -C=, thereby forming a compound of formula I-B:
Figure imgf000019_0002
I-B
or a pharmaceutically acceptable salt thereof, wherein each of Ring A, L1, L2, R1, Rx, R2, R3, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00111] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is -N= Y2 =C(R3)- and Y3 is -N-, thereby forming a compound of formula I-C:
Figure imgf000019_0003
I-C
or a pharmaceutically acceptable salt thereof, wherein each of Ring A, L1, L2, R1, Rx, R2, R3, m, n and p is as defined above and described in embodiments herein, both singly and in combination
[00112] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is -NH-, Y2 =C(R3)-, Y3 is -C=, L1 is a covalent bond, Ring A is cyclohexylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula II:
Figure imgf000020_0001
II
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00113] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)=, Y3 is -C=, L1 is a covalent bond, Ring A is tetrahydropyranylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula III:
Figure imgf000020_0002
III
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00114] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)=, Y3 is -C=, L1 is a covalent bond, Ring A is piperidinylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula IV:
Figure imgf000020_0003
IV or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00115] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is -NH-, Y2 -N=, Y3 is -C=, L1 is a covalent bond, Ring A is cyclohexylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula V, :
Figure imgf000021_0001
V
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00116] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =Ν-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)=, Y3 is -C=, L1 is a covalent bond, Ring A is cyclohexylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula VI:
Figure imgf000021_0002
VI
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00117] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -N= Y1 is - H-, Y2 -C(R3)= and Y3 is -C=, L1 is a covalent bond, Ring A is cyclohexylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula VII:
Figure imgf000022_0001
VII
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00118] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -N= X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)= and Y3 is -C=, L1 is a covalent bond, Ring A is cyclohexylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula VIII:
Figure imgf000022_0002
VIII
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00119] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is -N= Y2 =C(R3)- and Y3 is -N-, L1 is a covalent bond, Ring A is cyclohexylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula IX:
Figure imgf000022_0003
IX
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00120] In certain embodiments, the present invention provides a compound of formula I, wherein X1 is =C(R2)-, X2 is -C(R2)=, X3 is -C(R2)=, Y1 is - H-, Y2 -C(R3)=, Y3 is -C=, L1 is a covalent bond, Ring A is cyclohexylenyl, and L2 is -N(R)C(0)- thereby forming a compound of formula X:
Figure imgf000023_0001
X
or a pharmaceutically acceptable salt thereof, wherein each of R1, Rx, R2, R3, R, m, n and p is as defined above and described in embodiments herein, both singly and in combination.
[00121] Exemplary compounds of the invention are set forth in Table 1, below.
Table 1. Exemplary Compounds
Figure imgf000024_0001
1-1 (rel)-(lS,3R) 1-2 (rel)-(lS,3R)
Figure imgf000024_0002
1-3 (rel)-(lS,3R) 1-5 (rel)-(lS,3R)
Figure imgf000024_0003
Figure imgf000024_0004
-8 or 1-9 1-9 or 1-8
Figure imgf000025_0001
-19 1-20
Figure imgf000026_0001
-21 (rel)-(lS,3R) 1-22 (rel)-(lS,3S)
Figure imgf000026_0002
-27 (rel)-(lS,3R) 1-28 (rel)-(lS,3R)
Figure imgf000027_0001
1-29 (rel)-(lS,3R)
Figure imgf000027_0002
1-31 (rel)-(lS,3R) 1-32 (rel)-(lS,3R)
Figure imgf000027_0003
1-33 (rel)-(lS,3R) 1-34 (rel)-(lS,3R)
Figure imgf000027_0004
1-35 (rel)-(lS,3R) 1-36 (rel)-(lS,3R)
Figure imgf000028_0001
 -43 (rel)-(lS,3R) 1-44 or 1-45
Figure imgf000029_0001
Figure imgf000029_0002
Figure imgf000029_0003
Figure imgf000029_0004
-51 1-54 (rel)-(3R,5R)
Figure imgf000030_0001
-60 1-61 or 1-62
Figure imgf000031_0001
Figure imgf000032_0001
Figure imgf000033_0001
32
Figure imgf000034_0001
Figure imgf000035_0001
Figure imgf000036_0001
Figure imgf000037_0001
[00122] Exemplary compounds of the invention are set forth in Table 2, below.
Figure imgf000039_0001

Figure imgf000040_0001
1-57 (rel)-(lS,3S)
[00123] In some embodiments, the present invention provides a compound set forth in Table 1, above, or a pharmaceutically acceptable salt thereof. In some embodiments, the present invention provides a compound set forth in Table 2, above, or a pharmaceutically acceptable salt thereof.
5. Uses, Formulation and Administration
Pharmaceutically acceptable compositions
[00124] According to another embodiment, the invention provides a composition comprising a compound of this invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in compositions of this invention is such that is effective to measurably inhibit MTHFD2, in a biological sample or in a patient. In certain embodiments, the amount of compound in compositions of this invention is such that is effective to measurably inhibit MTHFD2, in a biological sample or in a patient. In certain embodiments, a composition of this invention is formulated for administration to a patient in need of such composition. In some embodiments, a composition of this invention is formulated for oral administration to a patient.
[00125] The term "patient," as used herein, means an animal, preferably a mammal, and most preferably a human.
[00126] The term "pharmaceutically acceptable carrier, adjuvant, or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, di sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.
[00127] Compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.
[00128] For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
[00129] Pharmaceutically acceptable compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
[00130] Alternatively, pharmaceutically acceptable compositions of this invention may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.
[00131] Pharmaceutically acceptable compositions of this invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
[00132] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically- transdermal patches may also be used.
[00133] For topical applications, provided pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxy ethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
[00134] For ophthalmic use, provided pharmaceutically acceptable compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.
[00135] Pharmaceutically acceptable compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
[00136] Most preferably, pharmaceutically acceptable compositions of this invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of this invention are administered with food.
[00137] The amount of compounds of the present invention that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, provided compositions should be formulated so that a dosage of between 0.01 - 100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.
[00138] It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound of the present invention in the composition will also depend upon the particular compound in the composition.
Uses of Compounds and Pharmaceutically Acceptable Compositions
[00139] Compounds and compositions described herein are generally useful for the inhibition of MTHFD2.
[00140] The activity of a compound utilized in this invention as an inhibitor of MTHFD2, may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of MTHFD2. Alternate in vitro assays quantitate the ability of the inhibitor to bind to MTHFD2. Detailed conditions for assaying a compound utilized in this invention as an inhibitor of MTHFD2, are set forth in the Examples below.
[00141] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.
[00142] Provided compounds are inhibitors of MTHFD2 and are therefore useful for treating one or more disorders associated with activity of MTHFD2. Thus, in certain embodiments, the present invention provides a method for treating a MTHFD2-mediated disorder comprising the step of administering to a patient in need thereof a compound of the present invention, or pharmaceutically acceptable composition thereof.
[00143] As used herein, the terms "MTHFD2-mediated" disorders, diseases, and/or conditions as used herein means any disease or other deleterious condition in which MTHFD2, is known to play a role. Accordingly, another embodiment of the present invention relates to treating or lessening the severity of one or more diseases in which MTHFD2 is known to play a role.
[00144] The methods described herein include methods for the treatment of cancer in a subject. As used in this context, to "treat" means to ameliorate or improve at least one symptom or clinical parameter of the cancer. For example, a treatment can result in a reduction in tumor size or growth rate. A treatment need not cure the cancer or cause remission 100% of the time, in all subjects.
[00145] As described herein, the application of agents, e.g., inhibitory nucleic acids or small molecules, that inhibit SHMT2 or MTHFD2 reduces cancer cell proliferation and thus treat cancer in subjects. Thus, in some embodiments, the methods described herein include administering a therapeutically effective dose of one or more agents that inhibit a mitochondrial 1-carbon (1-C) pathway enzyme, e.g., SHMT2, MTHFD2, and/or MTHFD1L.
[00146] As used herein, the term "cancer" refers to cells having the capacity for autonomous growth, i.e., an abnormal state or condition characterized by rapidly proliferating cell growth. The term is meant to include all types of cancerous growths or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues, or organs, irrespective of histopathologic type or stage of invasiveness. The term "tumor" as used herein refers to cancerous cells, e.g., a mass of cancer cells.
[00147] Cancers that can be treated or diagnoses using the methods described herein include malignancies of the various organ systems, such as affecting lung, breast, thyroid, lymphoid, gastrointestinal, and genito-urinary tract, as well as adenocarcinomas which include malignancies such as most colon cancers, renal-cell carcinoma, prostate cancer and/or testicular tumors, non-small cell carcinoma of the lung, cancer of the small intestine and cancer of the esophagus. [00148] In some embodiments, the methods described herein are used for treating or diagnosing a carcinoma in a subject. The term "carcinoma" is art recognized and refers to malignancies of epithelial or endocrine tissues including respiratory system carcinomas, gastrointestinal system carcinomas, genitourinary system carcinomas, testicular carcinomas, breast carcinomas, prostatic carcinomas, endocrine system carcinomas, and melanomas. In some embodiments, the cancer is renal carcinoma or melanoma. Exemplary carcinomas include those forming from tissue of the cervix, lung, prostate, breast, head and neck, colon and ovary. The term also includes carcinosarcomas, e.g., which include malignant tumors composed of carcinomatous and sarcomatous tissues. An "adenocarcinoma" refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures.
[00149] The term "sarcoma" is art recognized and refers to malignant tumors of mesenchymal derivation.
[00150] In some embodiments, the cancers that are treated by the methods described herein are cancers that have increased levels of glycine uptake or an increased expression or activity of a mitochondrial 1-c enzyme (e.g., SHMT2, MTHFD2, and/or MTHFD1L) relative to normal tissues or to other cancers of the same tissues; methods known in the art and described herein can be used to identify those cancers. In some embodiments, the methods include obtaining a sample comprising cells of the cancer, determining the level of glycine uptake or protein, mRNA, or activity of one or more mitochondrial 1-c enzymes (e.g., SHMT2, MTHFD2, and/or MTHFD1L) in the sample, and administering a treatment as described herein (e.g., an antifolate or an agent that inhibits MTHFD2, e.g., ebselen). In some embodiments, the cancer is one that is shown herein to have increased levels of glycine uptake.
[00151] In some embodiments, the present invention provides a method for treating one or more disorders, diseases, and/or conditions wherein the disorder, disease, or condition includes, but is not limited to, a cellular proliferative disorder.
Cellular Proliferative Disorders
[00152] The present invention features methods and compositions for the diagnosis and prognosis of cellular proliferative disorders (e.g., cancer) and the treatment of these disorders by targeting MTHFD2. Cellular proliferative disorders described herein include, e.g., cancer, obesity, and proliferation-dependent diseases. Such disorders may be diagnosed using methods known in the art.
Cancer [00153] Cancers include, without limitation, leukemias (e.g., acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocyte leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain disease, and solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, meningioma, melanoma, neuroblastoma, and retinoblastoma). In some embodiments, the cancer is melanoma or breast cancer. In some embodiments, the cancer is non-small cell lung cancer or hepatocellular carcinoma.
Other Proliferative Diseases
[00154] Other proliferative diseases include, e.g., obesity, benign prostatic hyperplasia, psoriasis, abnormal keratinization, lymphoproliferative disorders (e.g., a disorder in which there is abnormal proliferation of cells of the lymphatic system), chronic rheumatoid arthritis, arteriosclerosis, restenosis, and diabetic retinopathy. Proliferative diseases that are hereby incorporated by reference include those described in U.S. Pat. Nos. 5,639,600 and 7,087,648.
[00155] Pharmaceutically acceptable compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated. In certain embodiments, the compounds of the invention may be administered orally or parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.
[00156] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
[00157] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.
[00158] Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
[00159] In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
[00160] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
[00161] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar—agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.
[00162] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
[00163] The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.
[00164] Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
[00165] According to one embodiment, the invention relates to a method of inhibiting MTHFD2 activity in a biological sample comprising the step of contacting said biological sample with a compound of this invention, or a composition comprising said compound.
[00166] The term "biological sample", as used herein, includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof. [00167] Another embodiment of the present invention relates to a method of inhibiting MTHFD2 in a patient comprising the step of administering to said patient a compound of the present invention, or a composition comprising said compound.
[00168] According to another embodiment, the invention relates to a method of inhibiting MTHFD2 activity in a patient comprising the step of administering to said patient a compound of the present invention, or a composition comprising said compound. In other embodiments, the present invention provides a method for treating a disorder mediated by MTHFD2 in a patient in need thereof, comprising the step of administering to said patient a compound according to the present invention or pharmaceutically acceptable composition thereof. Such disorders are described in detail herein.
[00169] Depending upon the particular condition, or disease, to be treated, additional therapeutic agents that are normally administered to treat that condition, may also be present in the compositions of this invention. As used herein, additional therapeutic agents that are normally administered to treat a particular disease, or condition, are known as "appropriate for the disease, or condition, being treated."
[00170] A compound of the current invention may also be used to advantage in combination with other antiproliferative compounds. Such antiproliferative compounds include, but are not limited to aromatase inhibitors; anti estrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active compounds; alkylating compounds; histone deacetylase inhibitors; compounds which induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antineoplastic antimetabolites; platin compounds; compounds targeting/decreasing a protein or lipid kinase activity and further anti-angiogenic compounds; compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase; gonadorelin agonists; anti-androgens; methionine aminopeptidase inhibitors; matrix metalloproteinase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematologic malignancies; compounds which target, decrease or inhibit the activity of Flt-3; Hsp90 inhibitors such as 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545), IPI-504, CNF 1010, CNF2024, CNF 1010 from Conforma Therapeutics; temozolomide (Temodal®); kinesin spindle protein inhibitors, such as SB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazine from CombinatoRx; MEK inhibitors such as ARRY142886 from Array BioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer and leucovorin. The term "aromatase inhibitor" as used herein relates to a compound which inhibits estrogen production, for instance, the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term includes, but is not limited to steroids, especially atamestane, exemestane and formestane and, in particular, non-steroids, especially aminoglutethimide, roglethimide, pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole, fadrozole, anastrozole and letrozole. Exemestane is marketed under the trade name Aromasin™. Formestane is marketed under the trade name Lentaron™. Fadrozole is marketed under the trade name Afema™. Anastrozole is marketed under the trade name Arimidex™. Letrozole is marketed under the trade names Femara™ or Femar™. Aminoglutethimide is marketed under the trade name Orimeten™. A combination of the invention comprising a chemotherapeutic agent which is an aromatase inhibitor is particularly useful for the treatment of hormone receptor positive tumors, such as breast tumors.
[00171] The term "antiestrogen" as used herein relates to a compound which antagonizes the effect of estrogens at the estrogen receptor level. The term includes, but is not limited to tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen is marketed under the trade name Nolvadex™. Raloxifene hydrochloride is marketed under the trade name Evista™. Fulvestrant can be administered under the trade name Faslodex™. A combination of the invention comprising a chemotherapeutic agent which is an antiestrogen is particularly useful for the treatment of estrogen receptor positive tumors, such as breast tumors.
[00172] The term "anti -androgen" as used herein relates to any substance which is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (Casodex™). The term "gonadorelin agonist" as used herein includes, but is not limited to abarelix, goserelin and goserelin acetate. Goserelin can be administered under the trade name Zoladex™.
[00173] The term "topoisom erase I inhibitor" as used herein includes, but is not limited to topotecan, gimatecan, irinotecan, camptothecian and its analogues, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148. Irinotecan can be administered, e.g. in the form as it is marketed, e.g. under the trademark Camptosar™. Topotecan is marketed under the trade name Hycamptin™.
[00174] The term "topoisom erase II inhibitor" as used herein includes, but is not limited to the anthracyclines such as doxorubicin (including liposomal formulation, such as Caelyx™), daunorubicin, epirubicin, idarubicin and nemorubicin, the anthraquinones mitoxantrone and losoxantrone, and the podophillotoxines etoposide and teniposide. Etoposide is marketed under the trade name Etopophos™. Teniposide is marketed under the trade name VM 26- Bristol Doxorubicin is marketed under the trade name Acriblastin™ or Adriamycin™. Epirubicin is marketed under the trade name Farmorubicin™. Idarubicin is marketed, under the trade name Zavedos™. Mitoxantrone is marketed under the trade name Novantron.
[00175] The term "microtubule active agent" relates to microtubule stabilizing, microtubule destabilizing compounds and microtublin polymerization inhibitors including, but not limited to taxanes, such as paclitaxel and docetaxel; vinca alkaloids, such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate, and vinorelbine; discodermolides; cochicine and epothilones and derivatives thereof. Paclitaxel is marketed under the trade name Taxol™. Docetaxel is marketed under the trade name Taxotere™. Vinblastine sulfate is marketed under the trade name Vinblastin R.P™. Vincristine sulfate is marketed under the trade name Farmistin™.
[00176] The term "alkylating agent" as used herein includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel). Cyclophosphamide is marketed under the trade name Cyclostin™. Ifosfamide is marketed under the trade name Holoxan™.
[00177] The term "histone deacetylase inhibitors" or "FIDAC inhibitors" relates to compounds which inhibit the histone deacetylase and which possess antiproliferative activity. This includes, but is not limited to, suberoylanilide hydroxamic acid (SAHA).
[00178] The term "antineoplastic antimetabolite" includes, but is not limited to, 5- fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds, such as 5- azacytidine and decitabine, methotrexate and edatrexate, and folic acid antagonists such as pemetrexed. Capecitabine is marketed under the trade name Xeloda™. Gemcitabine is marketed under the trade name Gemzar™.
[00179] The term "platin compound" as used herein includes, but is not limited to, carboplatin, cis-platin, cisplatinum and oxaliplatin. Carboplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark Carboplat™. Oxaliplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark Eloxatin™.
[00180] The term "compounds targeting/decreasing a protein or lipid kinase activity; or a protein or lipid phosphatase activity; or further anti-angiogenic compounds" as used herein includes, but is not limited to, protein tyrosine kinase and/or serine and/or threonine kinase inhibitors or lipid kinase inhibitors, such as a) compounds targeting, decreasing or inhibiting the activity of the platelet-derived growth factor-receptors (PDGFR), such as compounds which target, decrease or inhibit the activity of PDGFR, especially compounds which inhibit the PDGF receptor, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib, SU101, SU6668 and GFB-111; b) compounds targeting, decreasing or inhibiting the activity of the fibroblast growth factor-receptors (FGFR); c) compounds targeting, decreasing or inhibiting the activity of the insulin-like growth factor receptor I (IGF-IR), such as compounds which target, decrease or inhibit the activity of IGF-IR, especially compounds which inhibit the kinase activity of IGF-I receptor, or antibodies that target the extracellular domain of IGF-I receptor or its growth factors; d) compounds targeting, decreasing or inhibiting the activity of the Trk receptor tyrosine kinase family, or ephrin B4 inhibitors; e) compounds targeting, decreasing or inhibiting the activity of the Axl receptor tyrosine kinase family; f) compounds targeting, decreasing or inhibiting the activity of the Ret receptor tyrosine kinase; g) compounds targeting, decreasing or inhibiting the activity of the Kit/SCFR receptor tyrosine kinase, such as imatinib; h) compounds targeting, decreasing or inhibiting the activity of the C-kit receptor tyrosine kinases, which are part of the PDGFR family, such as compounds which target, decrease or inhibit the activity of the c-Kit receptor tyrosine kinase family, especially compounds which inhibit the c-Kit receptor, such as imatinib; i) compounds targeting, decreasing or inhibiting the activity of members of the c-Abl family, their gene-fusion products (e.g. BCR-Abl kinase) and mutants, such as compounds which target decrease or inhibit the activity of c-Abl family members and their gene fusion products, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; or dasatinib (BMS-354825); j) compounds targeting, decreasing or inhibiting the activity of members of the protein kinase C (PKC) and Raf family of serine/threonine kinases, members of the MEK, SRC, JAK/pan-JAK, FAK, PDK1, PKB/Akt, Ras/MAPK, PI3K, SYK, TYK2, BTK and TEC family, and/or members of the cyclin-dependent kinase family (CDK) including staurosporine derivatives, such as midostaurin; examples of further compounds include UCN- 01, safingol, BAY 43-9006, Bryostatin 1, Perifosine; llmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521; LY333531/LY379196; isochinoline compounds; FTIs; PD 184352 or QAN697 (a P13K inhibitor) or AT7519 (CDK inhibitor); k) compounds targeting, decreasing or inhibiting the activity of protein-tyrosine kinase inhibitors, such as compounds which target, decrease or inhibit the activity of protein-tyrosine kinase inhibitors include imatinib mesylate (Gleevec™) or tyrphostin such as Tyrphostin A23/RG-50810; AG 99; Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; Tyrphostin B44 (+) enantiomer; Tyrphostin AG 555; AG 494; Tyrphostin AG 556, AG957 and adaphostin (4-{[(2,5- dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester; NSC 680410, adaphostin); 1) compounds targeting, decreasing or inhibiting the activity of the epidermal growth factor family of receptor tyrosine kinases (EGFRi ErbB2, ErbB3, ErbB4 as homo- or heterodimers) and their mutants, such as compounds which target, decrease or inhibit the activity of the epidermal growth factor receptor family are especially compounds, proteins or antibodies which inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3 and ErbB4 or bind to EGF or EGF related ligands, CP 358774, ZD 1839, ZM 105180; trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl-1033, EKB-569, GW-2016, El . l, E2.4, E2.5, E6.2, E6.4, E2. l l, E6.3 or E7.6.3, and 7H-pyrrolo-[2,3-d]pyrimidine derivatives; m) compounds targeting, decreasing or inhibiting the activity of the c-Met receptor, such as compounds which target, decrease or inhibit the activity of c-Met, especially compounds which inhibit the kinase activity of c-Met receptor, or antibodies that target the extracellular domain of c-Met or bind to HGF, n) compounds targeting, decreasing or inhibiting the kinase activity of one or more JAK family members (JAK1/JAK2/JAK3/TYK2 and/or pan-JAK), including but not limited to PRT- 062070, SB-1578, baricitinib, pacritinib, momelotinib, VX-509, AZD-1480, TG-101348, tofacitinib, and ruxolitinib; o) compounds targeting, decreasing or inhibiting the kinase activity of PI3 kinase (PI3K) including but not limited to ATU-027, SF-1126, DS-7423, PBI- 05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib; and; and q) compounds targeting, decreasing or inhibiting the signaling effects of hedgehog protein (Hh) or smoothened receptor (SMO) pathways, including but not limited to cyclopamine, vismodegib, itraconazole, erismodegib, and IPI-926 (saridegib).
[00181] The term "PI3K inhibitor" as used herein includes, but is not limited to compounds having inhibitory activity against one or more enzymes in the phosphatidylinositol-3 -kinase family, including, but not limited to ΡΒΚα, ΡΒΚγ, ΡΒΚδ, ΡΒΚβ, PBK-C2a, PBK-C2p, PBK-C2y, Vps34, pl l0-a, ρ110-β, ρ110-γ, ρ110-δ, p85-a, ρ85-β, ρ55-γ, ρ150, plOl, and ρ87. Examples of ΡΒΚ inhibitors useful in this invention include but are not limited to ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib.
[00182] The term "Bcl-2 inhibitor" as used herein includes, but is not limited to compounds having inhibitory activity against B-cell lymphoma 2 protein (Bcl-2), including but not limited to ABT-199, ABT-731, ABT-737, apogossypol, Ascenta's pan-Bcl-2 inhibitors, curcumin (and analogs thereof), dual Bcl-2/Bcl-xL inhibitors (Infinity Pharmaceuticals/Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and analogs thereof; see WO2008118802), navitoclax (and analogs thereof, see US7390799), NH-1 (Shenayng Pharmaceutical University), obatoclax (and analogs thereof, see WO2004106328), S-001 (Gloria Pharmaceuticals), TW series compounds (Univ. of Michigan), and venetoclax. In some embodiments the Bcl-2 inhibitor is a small molecule therapeutic. In some embodiments the Bcl-2 inhibitor is a peptidomimetic.
[00183] The term "BTK inhibitor" as used herein includes, but is not limited to compounds having inhibitory activity against Bruton's Tyrosine Kinase (BTK), including, but not limited to AVL-292 and ibrutinib.
[00184] The term "SYK inhibitor" as used herein includes, but is not limited to compounds having inhibitory activity against spleen tyrosine kinase (SYK), including but not limited to PRT-062070, R-343, R-333, Excellair, PRT-062607, and fostamatinib
[00185] Further examples of BTK inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in
WO2008039218 and WO2011090760, the entirety of which are incorporated herein by reference.
[00186] Further examples of SYK inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2003063794, WO2005007623, and WO2006078846, the entirety of which are incorporated herein by reference.
[00187] Further examples of PI3K inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2004019973, WO2004089925, WO2007016176, US8138347, WO2002088112, WO2007084786, WO2007129161, WO2006122806, WO2005113554, and WO2007044729 the entirety of which are incorporated herein by reference.
[00188] Further examples of JAK inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2009114512, WO2008109943, WO2007053452, WO2000142246, and WO2007070514, the entirety of which are incorporated herein by reference.
[00189] Further anti-angiogenic compounds include compounds having another mechanism for their activity, e.g. unrelated to protein or lipid kinase inhibition e.g. thalidomide (Thalomid™) and TNP-470. [00190] Examples of proteasome inhibitors useful for use in combination with compounds of the invention include, but are not limited to bortezomib, disulfiram, epigallocatechin-3- gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912, CEP-18770, and MLN9708.
[00191] Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase are e.g. inhibitors of phosphatase 1, phosphatase 2A, or CDC25, such as okadaic acid or a derivative thereof.
[00192] Compounds which induce cell differentiation processes include, but are not limited to, retinoic acid, α- γ- or δ- tocopherol or a- γ- or δ-tocotrienol.
[00193] The term cyclooxygenase inhibitor as used herein includes, but is not limited to, Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acid and derivatives, such as celecoxib (Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib or a 5-alkyl-2- arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid, lumiracoxib.
[00194] The term "bisphosphonates" as used herein includes, but is not limited to, etridonic, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acid. Etridonic acid is marketed under the trade name Didronel™. Clodronic acid is marketed under the trade name Bonefos™. Tiludronic acid is marketed under the trade name Skelid™. Pamidronic acid is marketed under the trade name Aredia™. Alendronic acid is marketed under the trade name Fosamax™. Ibandronic acid is marketed under the trade name Bondranat™. Risedronic acid is marketed under the trade name Actonel™. Zoledronic acid is marketed under the trade name Zometa™. The term "mTOR inhibitors" relates to compounds which inhibit the mammalian target of rapamycin (mTOR) and which possess antiproliferative activity such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578.
[00195] The term "heparanase inhibitor" as used herein refers to compounds which target, decrease or inhibit heparin sulfate degradation. The term includes, but is not limited to, PI-88. The term "biological response modifier" as used herein refers to a lymphokine or interferons.
[00196] The term "inhibitor of Ras oncogenic isoforms", such as H-Ras, K-Ras, or N-Ras, as used herein refers to compounds which target, decrease or inhibit the oncogenic activity of Ras; for example, a "farnesyl transferase inhibitor" such as L-744832, DK8G557 or Rl 15777 (Zarnestra™). The term "telomerase inhibitor" as used herein refers to compounds which target, decrease or inhibit the activity of telomerase. Compounds which target, decrease or inhibit the activity of telomerase are especially compounds which inhibit the telomerase receptor, such as telomestatin. [00197] The term "methionine aminopeptidase inhibitor" as used herein refers to compounds which target, decrease or inhibit the activity of methionine aminopeptidase. Compounds which target, decrease or inhibit the activity of methionine aminopeptidase include, but are not limited to, bengamide or a derivative thereof.
[00198] The term "proteasome inhibitor" as used herein refers to compounds which target, decrease or inhibit the activity of the proteasome. Compounds which target, decrease or inhibit the activity of the proteasome include, but are not limited to, Bortezomib (Velcade™) and MLN 341.
[00199] The term "matrix metalloproteinase inhibitor" or ("MMP" inhibitor) as used herein includes, but is not limited to, collagen peptidomimetic and nonpeptidomimetic inhibitors, tetracycline derivatives, e.g. hydroxamate peptidomimetic inhibitor batimastat and its orally bioavailable analogue marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551) BMS-279251 , BAY 12-9566, TAA211 , MMI270B or AAJ996.
[00200] The term "compounds used in the treatment of hematologic malignancies" as used herein includes, but is not limited to, FMS-like tyrosine kinase inhibitors, which are compounds targeting, decreasing or inhibiting the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, Ι-β-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, which are compounds which target, decrease or inhibit anaplastic lymphoma kinase.
[00201] Compounds which target, decrease or inhibit the activity of FMS-like tyrosine kinase receptors (Flt-3R) are especially compounds, proteins or antibodies which inhibit members of the Flt-3R receptor kinase family, such as PKC412, midostaurin, a staurosporine derivative, SU11248 and MLN518.
[00202] The term "HSP90 inhibitors" as used herein includes, but is not limited to, compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90; degrading, targeting, decreasing or inhibiting the HSP90 client proteins via the ubiquitin proteosome pathway. Compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90 are especially compounds, proteins or antibodies which inhibit the ATPase activity of HSP90, such as 17-allylamino, 17-demethoxygeldanamycin (17AAG), a geldanamycin derivative; other geldanamycin related compounds; radicicol and FIDAC inhibitors.
[00203] The term "antiproliferative antibodies" as used herein includes, but is not limited to, trastuzumab (Herceptin™), Trastuzumab-DMl, erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PR064553 (anti-CD40) and 2C4 Antibody. By antibodies is meant intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least 2 intact antibodies, and antibodies fragments so long as they exhibit the desired biological activity.
[00204] For the treatment of acute myeloid leukemia (AML), compounds of the current invention can be used in combination with standard leukemia therapies, especially in combination with therapies used for the treatment of AML. In particular, compounds of the current invention can be administered in combination with, for example, farnesyl transferase inhibitors and/or other drugs useful for the treatment of AML, such as Daunorubicin, Adriamycin, Ara-C, VP- 16, Teniposide, Mitoxantrone, Idarubicin, Carboplatinum and PKC412.
[00205] Other anti-leukemic compounds include, for example, Ara-C, a pyrimidine analog, which is the 2 -alpha-hydroxy ribose (arabinoside) derivative of deoxycytidine. Also included is the purine analog of hypoxanthine, 6-mercaptopurine (6-MP) and fludarabine phosphate. Compounds which target, decrease or inhibit activity of hi stone deacetylase (HDAC) inhibitors such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA) inhibit the activity of the enzymes known as histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), Trichostatin A and compounds disclosed in US 6,552,065 including, but not limited to, N-hydroxy-3-[4-[[[2-(2-methyl-lH- indol-3-yl)-ethyl]- amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(lH-indol-3-yl)ethyl]- amino]methyl]phenyl]-2E-2- propenamide, or a pharmaceutically acceptable salt thereof, especially the lactate salt. Somatostatin receptor antagonists as used herein refer to compounds which target, treat or inhibit the somatostatin receptor such as octreotide, and SOM230. Tumor cell damaging approaches refer to approaches such as ionizing radiation. The term "ionizing radiation" referred to above and hereinafter means ionizing radiation that occurs as either electromagnetic rays (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4th Edition, Vol. 1 , pp. 248-275 (1993).
[00206] Also included are EDG binders and ribonucleotide reductase inhibitors. The term "EDG binders" as used herein refers to a class of immunosuppressants that modulates lymphocyte recirculation, such as FTY720. The term "ribonucleotide reductase inhibitors" refers to pyrimidine or purine nucleoside analogs including, but not limited to, fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6- mercaptopurine (especially in combination with ara-C against ALL) and/or pentostatin. Ribonucleotide reductase inhibitors are especially hydroxyurea or 2-hydroxy-lH-isoindole-l ,3-dione derivatives.
[00207] Also included are in particular those compounds, proteins or monoclonal antibodies of VEGF such as l-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, l-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate; Angiostatin™; Endostatin™; anthranilic acid amides; ZD4190; ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies, such as rhuMAb and RHUFab, VEGF aptamer such as Macugon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibody, Angiozyme (RPI 4610) and Bevacizumab (Avastin™).
[00208] Photodynamic therapy as used herein refers to therapy which uses certain chemicals known as photosensitizing compounds to treat or prevent cancers. Examples of photodynamic therapy include treatment with compounds, such as Visudyne™ and porfimer sodium.
[00209] Angiostatic steroids as used herein refers to compounds which block or inhibit angiogenesis, such as, e.g., anecortave, triamcinolone, hydrocortisone, 11-a-epihydrocotisol, cortex ol one, 17a-hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone and dexamethasone.
[00210] Implants containing corticosteroids refers to compounds, such as fluocinolone and dexamethasone.
[00211] Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds or compounds with other or unknown mechanism of action.
[00212] The structure of the active compounds identified by code numbers, generic or trade names may be taken from the actual edition of the standard compendium "The Merck Index" or from databases, e.g. Patents International (e.g. EVIS World Publications).
[00213] A compound of the current invention may also be used in combination with known therapeutic processes, for example, the administration of hormones or radiation. In certain embodiments, a provided compound is used as a radiosensitizer, especially for the treatment of tumors which exhibit poor sensitivity to radiotherapy. [00214] A compound of the current invention can be administered alone or in combination with one or more other therapeutic compounds, possible combination therapy taking the form of fixed combinations or the administration of a compound of the invention and one or more other therapeutic compounds being staggered or given independently of one another, or the combined administration of fixed combinations and one or more other therapeutic compounds. A compound of the current invention can besides or in addition be administered especially for tumor therapy in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or a combination of these. Long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies, as described above. Other possible treatments are therapy to maintain the patient's status after tumor regression, or even chemopreventive therapy, for example in patients at risk.
[00215] Those additional agents may be administered separately from an inventive compound-containing composition, as part of a multiple dosage regimen. Alternatively, those agents may be part of a single dosage form, mixed together with a compound of this invention in a single composition. If administered as part of a multiple dosage regime, the two active agents may be submitted simultaneously, sequentially or within a period of time from one another normally within five hours from one another.
[00216] As used herein, the term "combination," "combined," and related terms refers to the simultaneous or sequential administration of therapeutic agents in accordance with this invention. For example, a compound of the present invention may be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present invention provides a single unit dosage form comprising a compound of the current invention, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.
[00217] The amount of both an inventive compound and additional therapeutic agent (in those compositions which comprise an additional therapeutic agent as described above) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Preferably, compositions of this invention should be formulated so that a dosage of between 0.01 - 100 mg/kg body weight/day of an inventive compound can be administered.
[00218] In those compositions which comprise an additional therapeutic agent, that additional therapeutic agent and the compound of this invention may act synergistically. Therefore, the amount of additional therapeutic agent in such compositions will be less than that required in a monotherapy utilizing only that therapeutic agent. In such compositions a dosage of between 0.01 - 1,000 μg/kg body weight/day of the additional therapeutic agent can be administered.
[00219] The amount of additional therapeutic agent present in the compositions of this invention will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent.
[00220] The compounds of this invention, or pharmaceutical compositions thereof, may also be incorporated into compositions for coating an implantable medical device, such as prostheses, artificial valves, vascular grafts, stents and catheters. Vascular stents, for example, have been used to overcome restenosis (re-narrowing of the vessel wall after injury). However, patients using stents or other implantable devices risk clot formation or platelet activation. These unwanted effects may be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition comprising a kinase inhibitor. Implantable devices coated with a compound of this invention are another embodiment of the present invention.
EXEMPLIFICATION
[00221] As depicted in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It will be appreciated that, although the general methods depict the synthesis of certain compounds of the present invention, the following general methods, and other methods known to one of ordinary skill in the art, can be applied to all compounds and subclasses and species of each of these compounds, as described herein.
[00222] List of abbrevations used in the experimental section.
AcOH: acetic acid
ACN: acetonitrile
Anhyd: anhydrous
Aq: aqueous
Bn: benzyl
Boc: tert-butoxycarbonyl
d: days
DBU: l,8-diazobicyclo[5.4.0]undec-7-ene
DCE: 1,2-dichloroethane
DCM: dichloromethane
DEA: diethylamine DIPEA: N,N-diisopropylethylamine
DMA: N,N-dimethylacetamide
DMAP: 4-dimethylaminopyridine
DMF: N,N-dimethylformamide
DMSO-dimethyl sulfoxide
DPPA: diphenylphosphoryl azide
EDC: l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
ee: enantiomeric excess
ESI: electrospray ionization
EtOAc: ethyl acetate
EtOH: ethanol
Fmoc: fluorenylmethyloxycarbonyl
Fmoc-OSu: N-(9-fluorenylmethoxycarbonyloxy)succinimide
h: hours
HATU: Ν,Ν,Ν' ,Ν' -tetramethyl-0-(7-azabenzotriazol- 1 -yl)uranium
hexafluorophosphate
HPLC: high performance liquid chromatography
IP A: isopropyl alcohol
M: molar
MeOH: methanol
min: minutes
mL: milliliters
mM: millimolar
mmol: millimoles
MTBE: methyl tert-butyl ether
MP: N-methylpyrrolidine
NMR: Nuclear Magnetic Resonance
°C: degrees Celsius
PBS: phosphate buffered saline
PE: petroleum ether
PPh3: triphenylphosphine
Rel: relative
R.T.: room temperature
sat: saturated
SFC: supercritical fluid chromatography
tBuOK: potassium tert-butoxide
TEA: triethylamine
Tf: trifluoromethanesulfonate
TfAA: trifluoromethanesulfonic anhydride
TFA: trifluoracetic acid
TIPS: triisopropylsilyl
THF: tetrahydrofuran
TLC: thin layer chromatography
pTSA: para-toluenesulfonic acid
[00224] Many compounds of the invention were made according to the general synthetic scheme shown in Scheme 1.
[00225] Scheme 1
Figure imgf000063_0001
[00226] Preparation of representative non-limiting examples of the compounds of the invention are described below.
[00227] Exemplary Procedure 1
Intermediate 1: (±)-3- 5-methoxy-lH-indol-3-yl)cyclohexanone.
Figure imgf000063_0002
[00228] To a solution of 5-methoxy-lH-indole (40 mmol, 5.88 g), cyclohex-2-enone (120 mmol, 11.52 g) in CH3CN (30 mL), was added Sc(OTf)3 (4 mmol, 1.96 g). The reaction mixture was stirred at 30 °C for 3h under N2 atmosphere. The reaction was concentrated and purified by flash chromatography (silica gel, PE/EtOAc =10/3) to give the title compound. ESI: m/z: 244.2 (M+H)+.
[00229] Intermediate 2: (±)-3-(5-chloro-lH-indol-3-yl)cyclohexanone.
Figure imgf000063_0003
[00230] The title compound was synthesized in a similar fashion as Intermediate 1 using 5-chloro-lH-indole. The crude product was purified by flash chromatography (silica gel, 0-30% EtOAc/PE) to afford (±)-3-(5-chloro-lH-indol-3-yl)cyclohexanone. ESI: m/z: 248.1 (M+H)+.
[00231] Intermediate 3: (±)-3-(5-ethox -lH-indol-3-yl)cyclohexanone.
Figure imgf000064_0001
[00232] The title compound was synthesized in a similar fashion as Intermediate 1 using 5-ethoxy-lH-indole. The crude product was purified by flash chromatography (silica gel, 0-20% EtOAc/PE) to afford (±)-3-(5-ethoxy-lH-indol-3-yl)cyclohexanone. ESI: m/z: 258.3 (M+H)+.
[00233] Intermediate 4: (±)-3-(5-chloro-2-meth l-lH-indol-3-yl)cyclohexanone.
Figure imgf000064_0002
[00234] The title compound was synthesized in a similar fashion as Intermediate 1 using 5-chloro-2 methyl-lH-indole. The product was purified by flash chromatography (silica gel, EtOAc/PE = 1 :4) to afford the (±)-3-(5-chloro-2-methyl-lH-indol-3-yl)cyclohexanone. ESI: m/z: 262.2 (M+H)+.
[00235] Intermediate 5: (±)-3-(5-methox -7-methyl-lH-indol-3-yl)cyclohexanone.
Figure imgf000064_0003
[00236] The title compound was synthesized in a similar fashion as Intermediate 1 using 5-methoxy-7-methyl-lH-indole. The crude product was purified by chromatography on silica gel to obtain 1.2 g of 3-(5-methoxy-7-methyl-lH-indol-3-yl)cyclohexanone. ESI: m/z: 258.2 (M+H)+.
[00237] Intermediate 6: (±)-3-(5-methyl-lH-indol-3-yl)cyclohexanone.
Figure imgf000065_0001
[00238] The title compound was synthesized in a similar fashion as Intermediate 1 using 5-methyl-lH-indole and Bi(N03)3 H20. The product was purified by flash chromatography (silica gel, PE/EtOAc=10/3) to give (±)-3-(5-methyl-lH-indol-3-yl)cyclohexanone. ESI: m/z: 228.3 (M+H)+.
[00239] Intermediate 7: (±)-3-(6-(b nzyloxy)-5-methoxy-lH-indol-3-yl)cyclohexanone.
Figure imgf000065_0002
[00240] The title compound was synthesized in a similar fashion as Intermediate 1 using 6-(benzyloxy)-5-methoxy-lH-indole. The product was purified by chromatography (silica gel , PE/EtOAc=10/3) to obtain (±)-3-(6-(benzyloxy)-5-methoxy-lH-indol-3-yl)cyclohexanone. ESI: m/z: 350.1 (M+H)+.
[00241] Intermediate 8: (±)-5-(5-chloro-lH-indol-3-yl) dihydro-2H-pyran-3(4H)-one.
Figure imgf000065_0003
[00242] The title compound was synthesized in a similar fashion as intermediate 1 using 5- chloro-lH-indole, 2H-pyran-3(6H)-one, and Bi(N03)3 5H20. The crude product was purified by flash chromatography (silica gel, PE/EtOAC=5/2) to give (±)-5-(5-chloro-lH-indol-3-yl) dihydro-2H-pyran-3(4H)-one. ESI: m/z: 472.0 (M+H)+.
[00243] Intermediate 9: (±)-tert-butyl 3-(5-methoxy-lH-indol-3-yl)-5-oxopiperidine-l- carboxylate.
Figure imgf000065_0004
[00244] The title compound was synthesized in a similar fashion as Intermediate 1 using 5-chloro-lH-indole, tert-butyl 5-oxo-5, 6-dihydropyridine-l(2H)-carboxylate, and Bi(NO3)3 5H20. The crude product was purified by flash chromatography (silica gel, PE/EtOAc=5/3) to obtain (±)-tert-butyl 3-(5-methoxy-lH-indol-3-yl)-5-oxopiperidine-l- carboxylate. ESI: m/z: 245.1 (M+H)+.
[00245] Intermediate 10: ±)-3-(5-methoxy-l-methyl-lH-indol-3-yl)cyclohexanone.
Figure imgf000066_0001
[00246] A mixture of (±)-3-(5-methoxy-lH-indol-3-yl)cyclohexanone (1 g, 4.11 mmol), iodomethane (1.17 g, 8.22 mmol), NaOH (329 mg, 8.22 mmol) in DMF (20 mL) was stirred at R.T. overnight. The reaction mixture was treated with water (20 mL) and extracted with EtOAc (3 * 100 mL). The combined organic fractions were dried with Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, PE/EtOAc=20%) to give (±)-3-(5-methoxy-l-methyl-lH-indol-3-yl)cyclohexanone. ESI: m/z: 258.2 (M+H)+.
[00247] Exemplary Procedure 2
Intermediate 11: (±)-3- 5-methoxy-lH-indol-3-yl)cyclohexanamine.
Figure imgf000066_0002
[00248] A suspension of (±)-3-(5-Methoxy-lH-indol-3-yl)cyclohexanone (10 g, 41.2 mmol) and ammonium acetate (95 g, 1234.6 mmol) in acetic acid (10 mL) and dichloroethane (200 mL) was stirred at 30 °C for 1.5 h. Then sodium triacetoxyborohydride (35 g, 164.6 mmol) was added to the mixture. After stirring at 30 °C for 4.5h, the mixture was concentrated under reduced pressure and the pH was adjusted to 8-9 with a saturated solution of sodium bicarbonate and extracted with DCM (3 * 100 mL). The combined organic fractions were dried with Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, DCM/MeOH=10/l) to give (±)-3-(5-methoxy-lH-indol-3- yl)cyclohexanamine. ESI: m/z: 245.2 (M+H)+.
[00249] Intermediate 12: (±)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine.
Figure imgf000067_0001
[00250] The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-3-(5-chloro-lH-indol-3-yl)cyclohexanone . The crude product was purified by flash chromatography (silica gel, 90:9: 1 DCM/MeOH/ H4OH) to give (±)-3-(5-chloro-lH-indol- 3-yl)cyclohexanamine. ESI: m/z: 249.2 (M+H)+.
[00251] Intermediate 13: (±)-3-(5-chloro-2-meth l-lH-indol-3-yl)cyclohexanamine.
Figure imgf000067_0002
[00252] The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-3-(5-chloro-2-methyl-lH-indol-3-yl)cyclohexanone. The crude product was purified by flash chromatography (silica gel, 90:9: 1 DCM/MeOH/NH4OH) to obtain (±)-3-(5-chloro-2- methyl-lH-indol-3-yl)cyclohexanamine. ESI: m/z: 263.2 (M+H)+.
[00253] Intermediate 14: (±)-3-(5- hoxy-lH-indol-3-yl)cyclohexanamine.
Figure imgf000067_0003
[00254] The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-3-(5-ethoxy-lH-indol-3-yl)cyclohexanone. The crude product was purified by flash chromatography (silica gel, 90:9: 1 DCM/MeOH/NH4OH) to obtain (±)-3-(5-ethoxy-lH- indol-3-yl)cyclohexanamine. ESI: m/z: 259.2 (M+H)+.
[00255] Intermediate 15: (±)-3-(5-methox -7-methyl-lH-indol-3-yl)cyclohexanamine.
Figure imgf000067_0004
[00256] The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-3-(5-methoxy-7-methyl-lH-indol-3-yl)cyclohexanone to provide (±)-3-(5-methoxy-7- methyl-lH-indol-3-yl)cyclohexanamine. ESI: m/z: 259.2 (M+H)+. [00257] Intermediate 16: (±)-3-(5-meth l-lH-indol-3-yl)cyclohexanamine.
Figure imgf000068_0001
[00258] The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-3-(5-methyl-lH-indol-3-yl)cyclohexanone. The product was purified by flash chromatography (silica gel, DCM/MeOH= 100/8) to give (±)-3-(5-methyl-lH-indol-3- yl)cyclohexanamine. ESI: m/z: 229.4 (M+H)+.
[00259] Intermediate 17: (±)-3-(5-m hoxy-l-methyl-lH-indol-3-yl)cyclohexanamine.
Figure imgf000068_0002
[00260] The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-3-(5-methoxy-l-methyl-lH-indol-3-yl)cyclohexanone to give(±)-3 -(5 -methoxy-1 -methyl - lH-indol-3-yl)cyclohexanamine. ESI: m/z: 259.3 (M+H)+.
[00261] Intermediate 18: (±)-3-(6-(benzyloxy)-5-methoxy-lH-indol-3- yl)cyclohexanamine.
Figure imgf000068_0003
The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-3-(6- (benzyloxy)-5-methoxy-lH-indol-3-yl)cyclohexanone. The crude product was purified by flash chromatography (silica gel, DCM/MeOH=10/l) to provide (±)-3-(6-(benzyloxy)-5- methoxy-lH-indol-3-yl)cyclohexanamine.ESI: m/z: 351.1 (M+H)+.
[00262] Intermediate 19: (±)-5-(5-chloro-lH-indol-3-yl) tetrahydro-2H-pyran-3 -amine.
Figure imgf000068_0004
[00263] The title compound was synthesized in a similar fashion as Intermediate 11 using (±)-5-(5-chloro-lH-indol-3-yl) dihydro-2H-pyran-3(4H)-one to give (±)-5-(5-chloro-lH- indol-3-yl) tetrahydro-2H-pyran-3 -amine. ESI: m/z: 251.1 (M+H)+.
[00264] Intermediate 20: (±)-Tert-butyl-3-amino-5-(5-methoxy-lH-indol-3-yl)piperidine- 1-carboxylate.
Figure imgf000069_0001
[00265] The title compound was made in a similar fashion as Intermediate 11 using (±)- tert-butyl 3-(5-methoxy-lH-indol-3-yl)-5-oxopiperidine-l-carboxylate to give (±)-tert-butyl 3-amino-5-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxylate as brown oil.
[00266] Exemplary Procedure 3
Intermediate 21: (±)-(9H-fluoren-9-yl) methyl rel-(l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexylcarbamate.
Figure imgf000069_0002
[00267] A mixture of (±)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine (20.4 mmol, 5 g), Fmoc-OSu (21.4 mmol, 7.22 g) and TEA (40.8 mmol, 4.15 g) in DCM (100 mL) was stirred for 2 h at RT. The reaction mixture was concentrated and purified by flash chromatography (silica gel) eluting first with PE/EtOAc=20/3, to remove the trans isomer, followed by PE/EtOAC=4/l to elute the cis isomer, (±)-(9H-fluoren-9-yl) methyl rel- (l S,3R)-3-(5- methoxy-lH-indol-3-yl)cyclohexylcarbamate. 1H- MR (CDC13, 400MHz) 57.87(s,lH), 7.77(d,2H), 7.62(d,2H), 7.40-7.26(m,6H), 7.07-7.03(m,2H), 6.89(dd, lH), 4.45(d,2H), 4.24(t,lH), 3.90(s,3H), 3.07(m, lH, Ha), 2.11 (m,lH, Hb), 2.10-1.64(m,5H), 1.40- 1.29(m,3H), ESI: m/z: 467.2 (M+H)+. Trans-isomer 1H- MR (CDC13, 400MHz) 57.84(s, lH), 7.77(d,2H), 7.60(d,2H), 7.40-7.06(m,6H), 7.06(d,lH), 6.92-6.84(m,2H), 4.40(d,2H), 4.22(t, lH), 3.87(s,3H), 2.92(m, lH, Ha), 2.41(m,lH, Hb), 2.11(m,lH), 1.91(m,lH), 1.39-1.17(m,6H), ESI: m/z: 467.2 (M+H)+. [00268] Intermediate 22: (±)-(9H-fluoren-9-yl) methyl rel-Q S,3R)-3-(5-chloro-lH-indol- 3-yl)cyclohexylcarbamate. i
Figure imgf000070_0001
[00269] The title compound was synthesized in a similar fashion as Intermediate 21 using (±)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine. The crude product was purified by flash chromatography (silica gel), first eluting with PE/EtOAc=5/l to provide the trans isomer. After the trans isomer was completely eluted the cis isomer was collected by eluting with PE/EtOAc=4/l to give (±)-(9H-fluoren-9-yl) methyl rel-(l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexylcarbamate. IH-NMR (CDC13, 400MHz) 58.02(s, lH), 7.78(d,2H), 7.62(m,4H), 7.43-7.26(m,5H), 7.15(d, lH), 6.96(s, lH), 4.42(d,2H), 4.23(t,lH), 2.88(m,lH, Ha), 2.36(m,lH, Hb), 2.11(m, lH), 1.91(m, lH), 1.39-1.17(m,6H), ESI: m/z: 472.0 (M+H)+.
[00270] Exemplary Procedure 4
Intermediate 23: (±)-rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine.
Figure imgf000070_0002
[00271] A mixture of (±)-(9H-fluoren-9-yl) methyl rel-(l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexylcarbamate (8.58 mmol , 4.00 g), and C2H8N2 (25.70 mmol , 1.54 g) in MeCN (50 mL) was stirred of 2 h at RT. The reaction was purified by flash chromatography (silica gel, DCM/MeOH=10/l) to give (±)-rel-(l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexanamine. ESI: m/z: 245.2 (M+H)+.
[00272] Intermediate 24: (±)-rel-(l S -3-(5-chloro-lH-indol-3-yl)cyclohexanamine.
Figure imgf000070_0003
[00273] The title compound was synthesized in a similar fashion as Intermediate 23 using (±)-(9H-fluoren-9-yl) methyl rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl carbamate. The product was purified by flash chromatography (silica gel, DCM/MeOH=10/l) to give :(±)-rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine. ESI: m/z: 249.1 (M+H)+.
[00274] Synthesis of Examples
Exemplary Procedure 5
Example 1: N-(rel)-(lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)cyclopentanecarboxamide [I-l].
Figure imgf000071_0001
[00275] A mixture of (±)-rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine (100 mg, 0.41mmol), cyclopentanecarboxylic acid (46.6 mg, 0.41 mmol), HATU (186.7 mg, 0.49 mmol) and DIPEA (158.6 mg, 1.23 mmol) in DMF (3 mL) was stirred for 2 h at 25 °C. The reaction mixture was poured into water (30 mL) and extracted with EtOAc (2 * 20 mL). The combined organics were washed with brine (3 * 30 mL), dried over Na2S04, concentrated, and purified by flash chromatography (silica gel, EtOAc/PE=l :2) to obtain N-(rel-(l S,3R)-3- (5-methoxy-lH-indol-3-yl)cyclohexyl)cyclopentanecarboxamide (I-l); ¾ NMR (400 MHz, DMSO^) δ 10.58 (s, 1H), 7.63 (d, J = 8.0 Hz, 1H), 7.20 (d, J = 8.8 Hz, 1H), 7.00 (dd, J = 5.6, 2.2 Hz, 2H), 6.70 (dd, J = 8.4, 2.2 Hz, 1H), 3.75 (s, 3H), 3.64-3.76 (m, 1H), 2.76-2.86 (m, 1H), 2.02-2.07 (m, 1H), 1.78-1.95 (m, 3H), 1.13-1.75 (m, 13H); ESI: m/z: 341.1 (M+H)+.
[00276] Example 2: N-(rel)-(lS,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)furan-2- carboxamide [1-2].
Figure imgf000071_0002
[00277] The title compound was synthesized in a similar fashion as Example 1 using furan 2-carboxylic acid and (±)-rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine to give N-(rel)-(l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)furan-2-carboxamide (1-2). ¾ NMR (400 MHz, DMSO^) δ 10.60 (s, 1H), 8.16-8.18 (d, 1H, J = 8.4 Hz), 7.81 (s, 1H, J = 8.8 Hz), 7.21-7.23 (d, 1H, J = 8.4 Hz), 7.09-7.10 (d, 1H, J = 3.6 Hz), 7.03 (d, 2H, J = 2.4 Hz), 6.70-6.72 (q, 1H, J = 2.4 Hz), 6.60 (q, 1H, J= 1.2 Hz), 3.95 (brs, 1H), 3.77 (s, 3H),2.87 (t, 1H), 2.07-2.12 (d, 1H, J = 12.8 Hz), 1.84-1.99 (m, 3H), 1.50-1.62 (m, 2H), 1.31-1.43 (m, 2H); ESI: m/z: 339.2 (M+H)+.
[00278] Example 3: N-(rel)-(lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrah dr o-2H-pyran-4-carboxamide [1-3] .
Figure imgf000072_0001
[00279] The title compound (1-3) was synthesized in a similar fashion as Example 1 using tetrahydro-2H-pyran-4-carboxylic acid and (±)-rel-(l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexanamine to give N-(rel)-(l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydro-2H-pyran-4-carboxamide (1-3). ¾ NMR (500 MHz, CDCb) δ 7.85 (b, 1H), 7.24 (d, J= 8.5 Hz, 1H), 7.06 (d, J = 2 Hz, 1H), 6.92 (d, J = 2 Hz, 1H), 6.86 (dd, J = 9 Hz, 2.5 Hz, 1H), 5.30 (d, J = 7.5 Hz, 1H), 4.02-3.99 (m, 3H), 3.87 (s, 3H), 3.49-3.48 (m, 1H), 3.42-3.36 (m, 2H), 2.96-2.92 (m, 1H), 2.36-2.25 (m, 2H), 2.09-2.06 (m, 2H), 1.94-1.89 (m, 1H), 1.79-1.74 (m, 4H), 1.43-1.12 (m, 3H); ESI: m/z: 357.1(M+H)+.
[00280] Example 4: (R)-N-(rel)-((lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-3-carboxamide [1-4].
Figure imgf000072_0002
[00281] (R)-Tetrahydrofuran-3-carbonyl chloride (165 mg, 1.23 mmol) was slowly added to a solution of ((±)-rel-(l S,3R)-3-(5-Methoxy-lH-indol-3-yl)cyclohexanamine (200 mg, 2.82 mmol), TEA (248 mg, 2.46 mmol) in DCM (20 mL) at 0 °C. Then the mixture was stirred at this temperature for 2h. The mixture was concentrated and the residue was purified by preparative HPLC using Xbridge OBD column eluting with 45%-75% acetonitrile in water (0.01% NH3 +10 mM NH4HC03) to give (R)-N-rel-((l S,3R)-3-(5-methoxy-lH-indol- 3-yl)cyclohexyl)tetrahydrofuran-3-carboxamide (1-4). ¾ NMR (500 MHz, DMSO-<f6) δ 10.58 (s, 1H), 7.84 (d, J = 7.8 Hz, 1H), 7.21 (d, J = 8.7 Hz, 1H), 7.01 (d, J = 7.3 Hz, 2H), 6.70 (dd, J= 8.7, 2.2 Hz, 1H), 3.82 (dd, J= 18.4, 8.2 Hz, 1H), 3.77 - 3.52 (m, 7H), 2.85 (m, J = 20.1, 9.6 Hz, 2H), 2.07 (s, 1H), 1.95 (m, J = 14.8, 7.3 Hz, 3H), 1.90 - 1.77 (m, 2H), 1.49 (d, J= 13.1 Hz, 1H), 1.39 - 1.27 (m, 2H), 1.18 (d, J= 12.3 Hz, 1H); ESI: m/z: 343.2 (M+H)+.
[00282] Example 5: N-(rel-(lS,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)oxazole-2- carboxamide [1-5].
Figure imgf000073_0001
[00283] The title compound was synthesized in a similar fashion as Example 1 using oxazole-2-carboxylic acid and (±)-(rel)-(l S,3R)-3-(5-Methoxy-lH-indol-3- yl)cyclohexanamine. The crude product was purified by preparative HPLC using a PHLEX ODS column eluting with 45%-65% acetonitrile in water (0.1%NH4 OH) to give N-(rel- (l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)oxazole-2-carboxamide (1-5). ¾ NMR (500 MHz, CDCb) δ 7.84 (b, 1H), 7.78 (s, 1H), 7.24-7.23 (m, 1H), 7.21 (s, 1H), 7.06 (d, J = 3.0 Hz, 1H), 6.96-6.93 (m, 2H), 6.84 (dd, J = 11.0, 3.0 Hz, 1H), 4.20-4.16 (m, 1H), 3.88 (s, 3H), 3.00-2.96 (m, 1H), 2.48-2.45 (m, 1H), 2.23-1.94 (m, 4H), 1.67-1.64 (m, 1H), 1.57-1.36 (m, 2H); ESI: m/z: 340.1 (M+H)+.
[00284] Examples 6, 7, 8, and 9: (S)-N-((lR,3S)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide and (R)-N-((lR,3S)-3-(5-methoxy- lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide [1-6 and 1-7] and (S)-N- ((lS,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide and (R)-N-((lS,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2- carboxamide [1-8 and 1-9].
Figure imgf000073_0002
1-6 1-7 1-8 1-9
[00285] A racemic mixture of the title compounds was synthesized in a similar fashion as example 4 using (±)-tetrahydro-2H-pyran-2-carbonyl chloride and (±)-rel-(l S,3R)-3-(5- methoxy-lH-indol-3-yl)cyclohexanamine. The product was purified by flash chromatography (silica gel, EtOAc/PE=l : l) to give (±)-N-(rel-(l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide. ESI: m/z: 257.1 (M+H)+.
[00286] The title compounds were resolved by chiral SFC using a CELRALPAK ID column eluting with Hexane/IPA/DEAO.1=70/30/0. l(v/v/v) and a CELRALPAK IF column eluting with Hexane/IPA/DEA0.1=85/15/0.1(v/v/v) to give Examples 6, 7, 8, and 9. (S)-N- ((lR,3S)-3-(5-Methoxy-lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide or (R)-N-(( 1 R, 3 S)-3 -(5 -Methoxy- 1 H-indol-3 -yl)cyclohexyl)tetrahydro-2H-pyran-2- carboxamide (1-6 ot 1-7), (retention time 13.58 min); ¾ MR (400 MHz, CDCb) δ 7.91 (br s, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.07 (d, J= 2.4 Hz, 1H), 6.92 (d, J= 2.4 Hz, 1H), 6.85 (dd, J = 8.8, 2.4 Hz, 1H), 6.45 (d, J = 8.4 Hz, 1H), 3.94-4.04 (m, 2H), 3.80 (s, 3H), 3.74-3.78 (m, 1H), 3.44-3.52 (m, 1H), 2.87-2.98 (m, 1H), 2.31-2.35 (m, 1H), 2.04-2.14 (m, 3H), 1.86-1.95 (m, 2H), 1.48-1.66 (m, 4H), 1.15-1.45 (m, 4H); ESI: m/z: 357.1 (M+H)+. (R)-N-((lR,3S)-3- (5-Methoxy-lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide or (S)-N- ((lR,3S)-3-(5-Methoxy-lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide (1-7 or 1-6), (retention time 15.48 min); ¾ MR (400 MHz, CDCb) δ 7.93 (br s, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.06 (d, J = 2.4 Hz, 1H), 6.91 (d, J = 2.0 Hz, 1H), 6.84 (dd, J = 8.8, 2.4 Hz, 1H), 6.47 (d, J = 8.4 Hz, 1H), 3.94-4.04 (m, 2H), 3.87 (s, 3H), 3.72-3.76 (m, 1H), 3.43-3.49 (m, 1H), 2.87-2.96 (m, 1H), 2.33-2.37 (m, 1H), 2.04-2.16 (m, 3H), 1.86-1.94 (m, 2H), 1.48- 1.66 (m, 4H), 1.14-1.42 (m, 4H); ESI: m/z: 357.1 (M+H)+. (S)-N-((l S,3R)-3-(5-Methoxy- lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide or (R)-N-((l S,3R)-3-(5- Methoxy-lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide (1-8 or 1-9), (retention time 13.34 min); ¾ NMR (400 MHz, CDCb) δ 7.91 (br s, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.07 (d, J = 2.4 Hz, 1H), 6.92 (d, J = 2.4 Hz, 1H), 6.85 (dd, J = 8.8, 2.4 Hz, 1H), 6.45 (d, J = 8.4 Hz, 1H), 3.94-4.04 (m, 2H), 3.80 (s, 3H), 3.74-3.78 (m, 1H), 3.44-3.52 (m, 1H), 2.87-2.98 (m, 1H), 2.31-2.35 (m, 1H), 2.04-2.14 (m, 3H), 1.86-1.95 (m, 2H), 1.48-1.66 (m, 4H), 1.15-1.45 (m, 4H); ESI: m/z: 357.1 (M+H)+. (R)-N-((l S,3R)-3-(5-Methoxy-lH- indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide or (S)-N-((l S,3R)-3-(5-Methoxy- lH-indol-3-yl)cyclohexyl)tetrahydro-2H-pyran-2-carboxamide (1-9 or 1-8), (retention time 14.57 min); ¾ NMR (400 MHz, CDCb) δ 7.96 (br s, 1H), 7.24 (d, J= 8.8 Hz, 1H), 7.05 (d, J = 2.4 Hz, 1H), 6.91 (d, J = 2.4 Hz, 1H), 6.84 (dd, J = 8.8, 2.4 Hz, 1H), 6.47 (d, J = 8.4 Hz, 1H), 3.94-4.04 (m, 2H), 3.87 (s, 3H), 3.72-3.76 (m, 1H), 3.43-3.49 (m, 1H), 2.87-2.96 (m, 1H), 2.33-2.37 (m, 1H), 2.04-2.16 (m, 3H), 1.86-1.94 (m, 2H), 1.48-1.66 (m, 4H), 1.14-1.42 (m, 4H); ESI: m/z: 357.2 (M+H)+. [00287] Example 10: (S)-(rel)-(lS,3R)-3-(5-methoxy-lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine-l-carbox late [I- 10].
Figure imgf000075_0001
(S)-Benzyl 2-(rel-(lS,3R)-3-(5-methoxy-lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine-l-carboxylate.
Figure imgf000075_0002
[00288] The title compound was synthesized in a similar fashion as Example 1 using (±)- rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine, and (S)-l-
(benzyloxycarbonyl)pyrrolidine-2-carboxylic acid. The crude product was purified by chromatography (silica gel, EtOAc/PE=l :2) to give (S)-benzyl 2-(rel-(l S,3R)-3-(5-methoxy- lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine-l-carboxylate. ESI: m/z: 476.2 (M+H)+. (S)-N-(Rel-(lS,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexylcarbamoyl)pyrrolidine-l- carboxylate [I- 10].
Figure imgf000075_0003
[00289] A mixture of (S)-benzyl 2-(rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine-l-carboxylate (210 mg, 0.44 mmol) and Pd/C (30 mg) in EtOH (10 mL) under H2 was stirred over 1 h at RT. The mixture was purified by chromatography on silica gel to give (S)-N-(rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl) cyclohexyl) pyrrolidine-2-carboxamide (1-10). ¾ NMR (500 MHz, DMSO-^) δ 10.62 (s, 1H), 7.45-7.31 (d, J = 8.0 Hz, 1H), 7.21-7.20 (d, J = 8.5 Hz, 1H), 7.02-7.00 (m, 2H), 6.71- 6.69 (m, 1H), 3.75-3.68 (m, 4H), 3.47-3.43 (m, 1H), 2.86-2.74 (m, 3H), 2.06-1.75 (m, 5H), 1.62-1.33 (m, 8H); ESI: m/z: 342.1 (M+H)+. [00290] Example 11: N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-2- methoxyacetamide [I-ll].
Figure imgf000076_0001
[00291] The title compound was synthesized in a similar fashion as Example 1 using 2- methoxyacetic acid and (±)-rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine. The crude product was purified by chromatography on silica gel to give N-(rel-(l S,3R)-3-(5- chloro-lH-indol-3-yl) cyclohexyl)-2-methoxyacetamide (I-ll). ¾ NMR (400 MHz, DMSO- d6) δ 10.98 (s, 1H), 7.62-7.58 (m, 2H), 7.35-7.33 (d, J = 8.4 Hz, 1H), 7.15-7.14 (d, J = 2.0 Hz, 1H), 7.06-7.03 (m, 1H), 3.86-3.77(m, 3H), 3.21(s, 3H), 2.89-2.82 (m, 1H), 2.00-1.79 (d, J = 13.6 Hz, 1H), 1.91-1.87 (d, J = 14.0 Hz, 1H), 1.82-1.79 (d, J = 13.2 Hz, 2H), 1.56-1.45(m, 2H), 1.39-1.31 (m, 2H); ESI: m/z: 321.1 (M+H) +.
[00292] Example 12: N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-2-hydroxy- 2-methylpropanamide [1-12].
Figure imgf000076_0002
[00293] The title compound was synthesized in a similar fashion as Example 1 using 2- hydroxy-2-methylpropanoic acid and (±)-rel-(l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexanamine. The crude product was purified by prep-TLC (silica gel, EtOAc/PE=l : l) to obtain N-(rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-2-hydroxy- 2-methylpropanamide (1-12). ¾ NMR (500 MHz, OMSO-d6) δ 10.97 (br s, 1H), 7.59 (d, J = 2.0 Hz, 1H), 7.32-7.36 (m, 2H), 7.16 (d, J = 2.0 Hz, 1H), 7.04 (dd, J = 8.5, 2.0 Hz, 1H), 5.32 (s, 1H), 3.67-3.76 (m, 1H), 2.82-2.89 (m, 1H), 1.77-2.00 (m, 4H), 1.44-1.55 (m, 2H), 1.26- 1.41 (m, 2H), 1.23 (s, 3H), 1.22 (s, 3H); ESI: m/z: 335.1 (M+H)+.
[00294] Example 13: N-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-2- hydroxyacetamide [1-13].
Figure imgf000077_0001
[00295] The title compound was synthesized in a similar fashion as Example 1 using 2- hydroxyacetic acid and (±)-rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine. The residue was purified by preparative HPLC using Xbridge OBD column eluting with 45%- 75% acetonitrile in water (0.01% NH3 +10 mM H4HC03) to give N-rel-((l S,3R)-3-(5- chloro-lH-indol-3-yl)cyclohexyl)-2-hydroxyacetamide (1-13). ¾ NMR (400 MHz, DMSO- d6) δ 10.99 (s, 1H), 7.58 (s, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.33 (d, J = 8.4 Hz, 1H), 7.15 (s, 1H), 7.04 (dd, J = 8.8, 2.0 Hz, 1H), 5.45-5.42 (m, 1H), 3.82 - 3.77 (m, 3H), 2.86- 2.84 (m, 1H), 2.02- 1.80 (m, 4H), 1.57 - 1.48 (m, 2H), 1.36 - 1.30 (m, 2H); ESI: m/z: 307.1 (M+H)+.
[00296] Example 14 and 15: (S)-N-((lR,3S)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-14] and (S)-N-((lS,3R)-3-(5-methoxy- lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-15].
Figure imgf000077_0002
1-14 T-15
[00297] The title compounds were synthesized in a similar fashion as Example 1 using (±)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2-carboxylic acid. The cis and trans isomers were separated by preparative HPLC using an Xbridge OBD column eluting with 35%-55% acetonitrile in water (0.01% NH3 +10 mM H4HC03) to provide the (S)-N-rel-((lR,3S)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide (cis isomers, retention time 6.5 min) and (S)-N-rel-((l S,3S)-3-(5-methoxy-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (trans isomers, retention time 7.0 min). The (S)-N-rel-((lR,3S)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide was separated into individual diastereomers by chiral SFC using a AD-H column eluting with 35% MeOH(0.1% H4OH) in carbon dioxide to give (S)-N-((lR,3S)-3- (5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (ee 98.8%, retention time 3.11 min) (1-14); ¾ NMR (400 MHz, CD3OD) δ 7.22 (d, J = 8.8 Hz, 1H), 7.07 (d, J = 2.0 Hz, 1H), 6.98 (s, 1H), 6.76 (dd, J = 2.4, 8.8 Hz, 1H), 4.28 (dd, J = 5.2, 8.0 Hz, 1H), 3.99- 3.81 (m, 6H), 2.95-2.88 (m, 1H), 2.30-2.18 (m, 2H), 2.07-2.03 (m, 1H), 1.97-1.81 (m, 5H), 1.65-1.30 (m, 4H); ESI: m/z: 343.2 (M+H)+; and (S)-N-((l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (ee 100%, retention time 6.29 min) (1-15); ¾ MR (400 MHz, CD3OD) δ 7.22 (d, J = 8.8 Hz, 1H), 7.07 (d, J = 2.4 Hz, 1H), 6.99 (s, 1H), 6.75 (dd, J = 2.0, 8.8 Hz, 1H), 4.28 (dd, J = 5.2, 8.4 Hz, 1H), 4.04-3.84 (m, 6H), 2.96-2.90 (m, 1H), 2.32-2.22 (m, 2H), 2.09-2.06 (m, 1H), 2.01-1.88 (m, 5H), 1.65-1.33 (m, 4H); ESI: m/z: 343.2 (M+H)+.
[00298] Example 16: N-(rel-(lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)acetamide [1-16].
Figure imgf000078_0001
[00299] To the solution of (±)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine (50 mg, 0.205 mmol) and Et3N (104 mg, 1.03 mmol) in dichloromethane (5 mL) was added acetic anhydride (62.8 mg, 0.615 mmol) at 0 °C. The mixture was stirred for 2 h at 25 °C. The reaction mixture was diluted in dichloromethane (30 mL) and washed with water (30 mL) and brine (30 mL), dried over Na2S04, concentrated at 40 °C and purified by Prep-TLC (silica gel, EtOAc/PE=l : l) to give N-(rel-(l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)acetamide (1-16). 1H MR (400 MHz, OMSO-d6) δ 7.89 (s, 1H), 7.23-7.26 (m, 1H), 7.06 (s, 1H), 6.91 (s, 1H), 6.85 (dd, J= 10.4, 2.0 Hz, 1H), 6.34 (d, J= 7.6 Hz, 1H), 3.95- 4.05 (m, 1H), 3.88 (s, 3H), 2.88-2.98 (m, 1H), 2.33-2.38 (m, 1H), 2.04-2.13 (m, 2H), 1.97 (s, 3H), 1.87-1.97 (m, 1H), 1.53-1.64 (m, 1H), 1.08-1.44 (m, 3H); ESI: m/z: 287.3 (M+H)+.
[00300] Example 17 and 18: (S)-N-((lR,3S)-3-(5-methoxy-l-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-17] and (S)-N-((lS,3R)-3-(5-methoxy- l-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-18].
Figure imgf000078_0002
1-17 1-18
[00301] The title compound was synthesized in a similar fashion as Example 1 using (±)- rel-3-(5-methoxy-l-methyl-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2- carboxylic acid. The mixture was purified by HPLC using Xbridge OBD column eluting with 45%-75% acetonitnle in water (0.01% H3 +10 mM NH4HC03). The diastereomers were separated by chiral SFC using an AD-H column eluting with 35% MeOH(0.1% H4OH) in carbon dioxide. (S)-N-((lR,3S)-3-(5-Methoxy-l-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-17), (retention time 3.85 min). ¾ MR (400 MHz, CD3OD) δ 7.07 (d, J= 8.8 Hz, 1H), 6.95 (d, J = 2.1 Hz, 1H), 6.80 - 6.62 (m, 2H), 4.15 (dd, J = 8.1, 5.6 Hz, 1H), 3.93 - 3.65 (m, 6H), 3.57 (s, 3H), 2.88 - 2.68 (m, 1H), 2.12- 1.90 (m, 2H), 1.97 - 1.65 (m, 6H), 1.54 - 1.19 (m, 4H); ESI: m/z: 357.1 (M+H)+. (S)-N- ((l S,3R)-3-(5-Methoxy-l-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-18), (retention time 6.56 min). ¾ NMR (400 MHz, CD3OD) δ 7.05 (d, J = 8.8 Hz, 1H), 6.94 (d, J = 2.2 Hz, 1H), 6.78 - 6.58 (m, 2H), 4.14 (dd, J = 8.2, 5.6 Hz, 1H), 3.93 - 3.63 (m, 6H), 3.54 (d, J = 13.5 Hz, 3H), 2.77 (ddd, J = 12.1, 9.0, 3.1 Hz, 1H), 2.19 - 2.00 (m, 2H), 1.96 - 1.68 (m, 6H), 1.53 - 1.15 (m, 4H); ESI : m/z: 357.1 (M+H)+
[00302] Example 19 and 20: (S)-N-((lR,3S)-3-(5-ethoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-19] and (S)-N-((lS,3R)-3-(5-ethoxy-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-20].
Figure imgf000079_0001
1-19 1-20
[00303] The title compounds were synthesized in a similar fashion as Example 1 using (±)-3-(5-ethoxy-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2-carboxylic acid. The cis and trans isomers were isolated by preparative HPLC using a PHLEX ODS column eluting with 34%-64% acetonitrile in water (0.1% NH3) to obtain (S)-N-(rel-(l S,3R)-3-(5- ethoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide, ESI: m/z: 357.3 (M+H)+ and (S)-N-(rel-(l S,3S)-3-(5-ethoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide ESI: m/z: 357.3 (M+H)+.
The (S)-N-(rel-(l S,3R)-3-(5-ethoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide was separated into individual diastereomers by chiral SFC using a AD column eluting with 40% MeOH (0.1% NH4OH) in carbon dioxide to give (S)-N-((lR,3S)-3-(5- ethoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-19), (ee 98.9%, retention time 2.77 min). ¾ NMR (400 MHz, CDC13) δ 7.84 (s, 1H), 7.22-7.26 (m, 1H), 7.09 (d, J = 2.8 Hz, 1H), 6.93 (d, J= 2.4 Hz, 1H), 6.84-6.87 (dd, J= 8.8, 2.0 Hz, 1H), 6.55 (d, J = 8.4 Hz, 1H), 4.32-4.36 (m, 1H), 4.07-4.13 (m, 2H), 3.96-4.02 (m, 1H), 3.82-3.89 (m, 2H), 2.89-2.97 (m, 1H), 2.24-2.32 (m, 2H), 1.99-2.10 (m, 3H), 1.78-1.95 (m, 3H), 1.53-1.64 (m, 1H), 1.45 (t, J = 7.2 Hz, 3H), 1.14-1.50 (m, 3H); ESI: m/z: 357.1 (M+H)+; and (S)-N-((l S,3R)-3-(5- ethoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-20), (ee 100.0%, retention time 5.47 min). ¾ NMR (400 MHz, CDCb) δ 7.83 (s, 1H), 7.22-7.24 (m, 1H), 7.07 (d, J = 2.4 Hz, 1H), 6.91 (d, J= 2.0 Hz, 1H), 6.83-6.87 (dd, J = 8.8, 2.4 Hz, 1H), 6.57 (d, J = 8.4 Hz, 1H), 4.31-4.35 (m, 1H), 4.07-4.13 (m, 2H), 3.84-4.03 (m, 3H), 2.88-2.97 (m, 1H), 2.24-2.38 (m, 2H), 2.00-2.13 (m, 3H), 1.81-2.02 (m, 3H), 1.53-1.66 (m, 1H), 1.44 (t, J = 7.0 Hz, 3H), 1.13-1.41 (m, 3H); ESI: m/z: 357.2 (M+H)+.
[00304] Example 21 and 22: (S)-N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-21] and (S)-N-(rel-(lS,3S)-3-(5-chloro- lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-22] .
Figure imgf000080_0001
1-21 1-22
[00305] The title compounds were synthesized in a similar fashion as Example 1 using (±)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2-carboxylic acid. The cis and trans diastereomers were separated by preparative HPLC using a PHLEX ODS column eluting with 34%-64% acetonitnle in water (0.1%NH3) to obtain (S)-N-(rel-(l S,3R)- 3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-21); ¾ NMR (400 MHz, CDCb) δ 8.05 (s, 1H), 7.60-7.62 (m, 1H), 7.25-7.28 (m, 1H), 7.10-7.14 (m, 1H), 6.94- 6.98 (m, 1H), 6.57-6.59 (m, 1H), 4.31-4.36 (m, 1H), 3.86-3.95 (m, 3H), 2.88-2.95 (m, 1H), 2.25-2.34 (m, 2H), 2.01-2.10 (m, 3H), 1.82-1.92 (m, 3H), 1.18-1.62 (m, 4H); ESI: m/z: 347.2 (M+H)+; and (S)-N-(rel-(l S,3 S)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide (1-22), ¾ NMR (400 MHz, CDCb) δ 8.09 (s, 1H), 7.58-7.51 (m, 1H), 7.26-7.29 (m, 1H), 7.07-7.14 (m, 2H), 6.98-7.01 (m, 1H), 4.31-4.41 (m, 2H), 3.90-4.04 (m, 2H), 3.05- 3.09 (m, 1H), 2.26-2.35 (m, 1H), 1.87-2.14 (m, 6H), 1.62-1.70 (m, 5H); ESI: m/z: 347.1 (M+H)+. [00306] Example 23 and 24: (S)-N-((lS,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-23] and (S)-N-((lR,3S)-3-(5-chloro-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-24].
Figure imgf000081_0001
1-23 1-24
[00307] (S)-N-(rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide (example 21) (160 mg, 0.46 mmol) was separated into individual diastereomers by chiral SFC using a WHELK-H column eluting with 40% MeOH (0.1% H4OH) in carbon dioxide. (S)-N-((l S,3R)-3-(5-Chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide (1-23), (ee 100%, retention time 4.41 min); ¾ NMR (400 MHz, CDCb) δ 8.02 (s, 1H), 7.60 (d, J = 2.0 Hz, 1H), 7.25-7.28 (m, 1H), 7.10-7.14 (m, 1H), 6.95 (m, 1H), 6.57- 6.59 (m, 1H), 4.31-4.36 (m, 1H), 3.86-3.95 (m, 3H), 2.88-2.95 (m, 1H), 2.34-2.25 (m, 2H), 2.01-2.10 (m, 3H), 1.82-1.92 (m, 3H), 1.18-1.62 (m, 4H); ESI: m/z: 347.1 (M+H) +. (S)-N- ((lR,3S)-3-(5-Chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-24) (ee 100%, retention time 5.72 min); ¾ MR (400 MHz, CDCb) δ 8.03 (s, 1H), 7.61 (d, J = 2.0 Hz, 1H), 7.25-7.28 (m, 1H), 7.10-7.14 (m, 1H), 6.97 (m, J = 2.0 Hz, 1H), 6.57-6.59 (m, 1H), 4.32-4.36 (m, 1H), 3.82-4.01 (m, 3H), 2.88-2.95 (m, 1H), 2.25-2.34 (m, 2H), 2.01-2.10 (m, 3H), 1.82-1.92 (m, 3H), 1.15-1.60 (m, 4H); ESI: m/z: 347.1 (M+H)+.
[00308] Example 25 and 26: (S)-N-((lS,3S)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)- tetrahydrofuran-2-carboxamide and (S)-N-((lR,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-25 and 1-26].
Figure imgf000081_0002
1-25 T-26
[00309] (S)-N-(rel-(l S,3S)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide (Example 22) (170 mg, 0.49 mmol) was separated into individual diastereomers by chiral SFC using an AD column eluting with 40% MeOH (0.5% H4OH) in carbon dioxide to give (S)-N-((l S,3S)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide or (S)-N-((lR,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide (1-25), (ee 95.5%, retention time 5.16 min); ¾ NMR (400 MHz, CDCb) δ 8.09 (s, 1H), 7.52 (d, J = 2.0 Hz, 1H), 7.26-7.29 (m, 1H), 7.07-7.14 (m, 2H), 7.01-7.03 (m, 1H), 4.30-4.39 (m, 2H), 3.90-4.04 (m, 2H), 3.00-3.06 (m, 1H), 2.29-2.35 (m, 1H), 1.87-2.14 (m, 6H), 1.58-1.73 (m, 5H); ESI: m/z: 347.1 (M+H)+, and (S)-N-((lR,3R)-3-(5-chloro-lH-indol- 3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (S)-N-((l S,3S)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-26) (ee 100%, retention time 4.20 min); ¾ MR (400 MHz, CDCb) δ 8.07 (s, 1H), 7.57 (d, J = 2.0 Hz, 1H), 7.26-7.28 (m, 1H), 7.07- 7.14 (m, 2H), 6.98-7.01 (m, 1H), 4.30-4.40 (m, 2H), 3.90-4.04 (m, 2H), 3.05-3.09 (m, 1H), 2.26-2.35 (m, 1H), 1.87-2.14 (m, 6H), 1.62-1.70 (m, 5H); ESI: m/z: 347.1 (M+H)+.
[00310] Example 27: (S)-N-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-N- methyltetrahydrofuran-2-carboxamid -27].
Figure imgf000082_0001
(S)-N-(Rel-(lS,3R)-3-(5-chloro-l-(triisopropylsilyl)-lH-indol-3-yl)cyclohexyl)-N- methyltetrahydrofuran-2-carboxamide.
Figure imgf000082_0002
[00311] To the suspension of NaH (115 mg, 2.88 mmol) in THF (2 mL) was added drop wise a solution of (S)-N-rel-((l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran- 2-carboxamide (Example 21) (100 mg, 0.29 mmol) in THF (2 mL) at 0 °C. The mixture was stirred for 0.5 h at 0 °C, then a solution of TIPSCI (56 mg, 0.29 mmol) was added drop wise and the mixture was stirred for 1 h at 25 °C. The mixture was cooled to 0 °C and Mel (122 mg, 0.86 mmol) was added. The resulting mixture was stirred for 1 h at 25 °C. The reaction mixture was poured into water (30 mL) and extracted with EtOAc (2 * 20 mL). The combined organics were washed with brine (30 mL), dried over Na2S04, concentrated, and purified by flash chromatography (silica gel, EtOAc/PE = 1 :5) to obtain (S)-N-(Rel-(l S,3R)- 3-(5-chloro-l-(triisopropylsilyl)-lH-indol-3-yl)cyclohexyl)-N-methyltetrahydrofuran-2- carboxamide.
(S)-N-Rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-N-methyltetrahydrofuran-2- carboxamide [1-27].
Figure imgf000083_0001
[00312] To a solution of (S)-N-(rel-(l S,3R)-3-(5-chloro-l-(triisopropylsilyl)-lH-indol-3- yl)cyclohexyl)-N-methyltetrahydrofuran-2-carboxamide (128 mg, 0.25 mmol) in THF (3 mL) was added tetrabutylammonium fluoride (0.3 mL, 0.30 mmol, 1 M in THF). The mixture was stirred for 1 h at 25 °C. The reaction mixture was partitioned between EtOAc (30 mL) and water (20 mL). The organic layer was washed with brine (30 mL), dried over Na2S04, concentrated, and purified by preparative HPLC using an Agela Durashell CI 8 (L) column eluting with 72%-82% acetonitrile in water (10 mM H4HCO3) to obtain (S)-N-rel-((l S,3R)- 3-(5-chloro-lH-indol-3-yl)cyclohexyl)-N-methyltetrahydrofuran-2-carboxamide (1-27). ¾ MR (500 MHz, CDCb) δ 8.10-8.16 (m, 1H), 7.55-7.60 (m, 1H), 7.25-7.30 (m, 1H), 7.10- 7.15 (m, 1H), 6.95-7.00 (m, 1H), 4.60-4.74 (m, 1.5H), 3.84-4.10 (m, 2.5H), 2.84-2.95 (m, 4H), 2.18-2.41 (m, 1H), 1.87-2.05 (m, 6H), 1.33-1.80 (m, 5H); ESI: m/z: 361.1 (M+H)+.
[00313] Example 28: 5-chloro-3-(rel-(lS,3R)-3-((S)-tetrahydrofuran-2-carboxamido)- cyclohexyl)-lH-indole-l-carboxami -28].
Figure imgf000083_0002
[00314] To a solution of (S)-N-(rel-(l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (Example 21) (50 mg, 0.14 mmol) in THF (0.5 mL) at 0 °C was added NaH (20 mg, 0.56 mmol). The mixture was stirred at 0 °C for 30 min before slow addition of chlorosulfonyl isocyanate (20 mg, 0.14 mmol) maintaining temp between 5 to 10 °C. The solution was warmed to rt and stirred for 3.5 h. Acetic acid (1.0 mL) was added and the solution was stirred at rt for 1.5 h before addition of ice cubes and water (10 mL). The suspension was stirred at rt for 30 min and the precipitate was collected by filtration. The solid was suspended in MeOH and the solid was collected by filtration. The crude solid was purified by prep-HPLC using XBridge OBD column eluting with 55-67% ACN in water (10 mM H4HCO3) to afford 5-chloro-3-(rel-(l S,3R)-3-((S)-tetrahydrofuran- 2-carboxamido)cyclohexyl)-lH-indole-l-carboxamide (1-28). ¾ NMR (400 MHz, CDCb) δ 8.14-8.10 (m, 1H), 7.58-7.55 (m, 1H), 7.31-7.27 (m, 1H), 7.13 (d, J = 8.4 Hz, 1H), 6.66-6.61 (m, 1H), 5.39 (br s, 1H), 5.35 (br s, 1H), 4.37-4.33 (m, 1H), 4.00-3.85 (m, 3H), 2.92-2.86 (m, 1H), 2.35-2.31 (m, 2H), 2.24-2.11 (m, 3H), 2.06-1.84 (m, 3H), 1.44-1.17 (m, 4H); ESI: m/z: 390.7(M+H)+.
[00315] Example 29: rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)-N-(((S)-tetrahydrofuran- 2-yl)methyl)cyclohexanamine [1-29] .
Figure imgf000084_0001
[00316] To a solution of (S)-N-(rel-(l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (Example 21) (100 mg, 0.29 mmol) in THF (4 ml) was added B2H5/THF (1M) (3 mL). The mixture was stirred at room temperature for 16 h under an Argon atmosphere. The mixture was concentrated in vacuo. HCl / methanol (4 N, 4 mL, 4 mmol) was added to the mixture. The mixture was stirred at 50 °C for 4 h. The mixture was diluted with EtOAc (100 ml), washed with saturated solution of sodium carbonate (3 χ 20 mL), dried (Na2S04) and concentrated. The residue was purified by preparative HPLC using a PHLEX ODS column eluting with 34%-64% acetonitrile in water (0.1% H4OH) to give rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)-N-(((S)-tetrahydrofuran-2- yl)methyl)cyclohexanamine (1-29). ¾ NMR (500 MHz, CDCb) δ 8.05 (b, 1H), 7.59 (m, 1H), 7.27-7.25 (m, 1H), 7.12 (dd, J = 11.0, 3.0 Hz, 1H), 6.97 (d, J = 3.0 Hz, 1H), 4.05-4.02 (m, 1H), 3.88-3.83 (m, 1H), 3.78-3.72 (m, 1H), 2.87-2.83 (m, 2H), 2.73-2.66 (m, 2H), 2.30-2.27 (m, 1H), 2.07-1.83 (m, 6H), 1.58-1.45 (m, 2H), 1.39-1.29 (m, 2H), 1.18-1.15 (m, 1H); ESI: m/z: 333.1 (M+H)+.
[00317] Example 30: rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)-N-(((R)-tetrahydrofuran- 2-yl)methyl)cyclohexanamine [1-30] .
Figure imgf000085_0001
[00318] The title compound was synthesized in a similar fashion as Example 24 using (R)- N-(rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (synthesized from (±)-rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine and (R)- tetrahydrofuran-2-carboxylic acid as in Example 15). The crude product was purified by preparative HPLC using a Xbridge OBD column eluting with 50%-70% acetonitrile in water (10 mM H4HCO3) to obtain rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)-N-(((R)- tetrahydrofuran-2-yl)methyl)cyclohexanamine (1-30). ¾ NMR (400 MHz, CDCh) δ 7.99 (br s, 1H), 7.59-7.61 (m, 1H), 7.24-7.28 (m, 1H), 7.13 (dd, J = 8.8, 2.0 Hz, 1H), 6.98 (d, J = 2.4 Hz, 1H), 3.96-4.05 (m, 1H), 3.82-3.88 (m, 1H), 3.72-3.78 (m, 1H), 2.77-2.87 (m, 2H), 2.63- 2.72 (m, 2H), 2.24-2.31 (m, 1H), 1.95-2.08 (m, 3H), 1.84-1.93 (m, 3H), 1.44-1.60 (m, 3H), 1.28-1.41 (m, 2H), 1.09-1.22 (m, 1H); ESI: m/z: 333.1 (M+H)+.
[00319] Example 31: N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)tetrahydro- 2H-pyran-4-amine [1-31].
Figure imgf000085_0002
[00320] A mixture of (±)-rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine (110 mg, 0.28 mmol) and dihydro-2H-pyran-4(3H)-one (88 mg, 0.88 mmol) in MeOH (8 mL) was stirred for 30 min at 25 °C. Then NaBH(OAc)3 (466 mg, 2.20 mmol) was added portion wise at 0 °C. The mixture was stirred for 16 h at 25 °C. The reaction was quenched with water (3 mL) and the mixture was concentrated. The residue was dissolved in EtOAc (40 mL), washed with aqueous NaHC03 (40 mL) and brine (40 mL), dried over Na2S04, concentrated at 40 °C and purified by preparative HPLC using a Xbridge OBD column eluting with 40%-60% acetonitrile in water (10 mM NH4HC03) to give N-(rel-(l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)tetrahydro-2H-pyran-4-amine (1-31). ¾ NMR (400 MHz, CDCb) δ 7.99 (br s, 1H), 7.60 (d, J = 2.0 Hz, 1H), 7.26-7.28 (m, 1H), 7.13 (dd, J = 8.4, 2.0 Hz, 1H), 6.99 (d, J = 2.0 Hz, 1H), 3.95-4.01 (m, 2H), 3.37-3.44 (m, 2H), 2.79-2.92 (m, 3H), 2.21-2.26 (m, 1H), 1.96-2.09 (m, 2H), 1.82-1.94 (m, 3H), 1.30-1.57 (m, 6H), 1.09-1.21 (m, 1H); ESI: m/z: 333.1 (M+H)+.
[00321] Example 32: (±)-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-3- phenylurea [1-32].
Figure imgf000086_0001
[00322] To a solution of (±)-rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine (1 mmol, 0.248 g), TEA (2.4 mmol, 0.242 g), benzoic acid (2 mmol, 0.244 g) in toluene (10 mL), was added DPPA (2.4 mmol, 0.660 g). The reaction mixture was stirred at 80 °C for 3h under N2 atmosphere. The mixture was treated with water (20 mL) and extracted with EtOAc (2 * 100 mL). The combined organic layers were dried with Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 1 : 1 EtOAc/PE) to give the crude product. The crude product was purified by preparative HPLC using a Xbridge OBD column eluting with 45%-75% acetonitrile in water (0.1% H4OH) to give (±)-rel- ((l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-3-phenylurea (1-32). ¾ MR (400 MHz, DMSO^) δ 10.99 (b, 1H), 8.31 (b, 1H), 7.60 (s, 1H), 7.33-7.38 (t, 3H), 7.17-7.22 (t, 3H), 7.03-7.06 (dd, J = 6.8 Hz, 2 Hz, 1H), 6.87 (t, 1H), 6.06-6.08 (d, J = 8 Hz, 1H), 3.63 (b, 1H), 2.88(m, 1H), 2.5(m, 1H), 1.81-1.94 (m, 3H), 1.51(m, 1H), 1.36(m, 2H), 1.17(m, 1H); ESI: m/z: 368.2 (M+H)+.
[00323] Example 33: (±)-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-3- methylurea [1-33].
Figure imgf000086_0002
[00324] The title compound was synthesized in a similar fashion as Example 27 using (±)- rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine and acetic acid. The product was purified by preparative HPLC using a Xbridge OBD column eluting with 45%-75% acetonitrile in water (0.1% H4OH) to give (±)- rel-((l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexyl)-3-methylurea (1-33). ¾ NMR (400 MHz, DMSO-d) δ 10.97 (s, 1H), 7.56 (d, 1H, J = 1.6 Hz), 7.32-7.34 (d, 1H, J = 8.4 Hz), 7.13 (d, 1H, J = 2.4 Hz), 7.02-7.04 (q, 1H, J = 2.0 Hz), 5.74-5.76 (d, 1H, J = 7.6 Hz), 5.58 (d, 1H, J = 4.4 Hz), 3.50 (brs, 1H), 2.79 (t, 1H), 2.51(d, 3H, J= 4.8 Hz), 2.08-2.08 (d, 1H, J = 9.0 Hz), 1.76-1.90 (m, 3H), 1.45-1.48 (m, 1H), 1.20-1.32 (m, 2H), 1.07-1.10 (m, 1H); ESI: m/z: 306.1 (M+H)+.
[00325] Example 34: (±)-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)-3- (pyridin-2-yl)urea [1-34].
Figure imgf000087_0001
[00326] The title compound was synthesized in a similar fashion as Example 27 using picolinic acid and (±)-rel-(l S,3R)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine. The product was purified by prep-HPLC using Xbridge OBD column eluting with 55-68% acetonitrile in water (0.01% H3 +10 mM H4HC03) to give (±)-rel-(Q S,3R)-3-(5-chloro- lH-indol-3-yl)cyclohexyl)-3-(pyridin-2-yl)urea (1-34). ¾ NMR (400 MHz, DMSO- ck) δ 10.99 (s, 1H), 9.11 (s, 1H), 8.15 (d, J = 4.8 Hz, 2H), 7.67-7.60 (m, 2H), 7.34-7.31 (m, 2H), 7.19 (s, 1H), 7.05-7.23 (m, 1H), 6.90-6.87 (m, 1H), 3.73-3.71 (m, 1H), 2.92-2.86 (m, 1H), 2.22-2.19 (m, 1H), 2.02-1.81 (m, 3H), 1.55-1.33 (m, 4H); ESI: m/z: 369.7 (M+H)+.
[00327] Example 35: (±)-rel-((lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl)urea [I- 35].
Figure imgf000087_0002
[00328] A solution of (±)-rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexanamine (100 mg, 0.40 mmol), and urea (242 mg, 4.0 mmol) in H20 (2 mL) was stirred at reflux for 16h. The residue was purified by preparative using Xbridge OBD column eluting with 45%-75% acetonitrile in water (0.01% H3 + 10 mM H4HC03) to give (±)-rel-((l S,3R)-3-(5-chloro- lH-indol-3-yl)cyclohexyl)urea (1-35). ¾ NMR (500 MHz, CD3OD) δ 7.58(d, J = 2 Hz, 1H), 7.29 (d, J = 7 Hz, 1H), 7.05 (m, 2H), 3.65 (m, 1H), 2.90-2.94 (m, 1H), 2.30 (d, J = 2 Hz, 1H), 2.02-2.05 (m, 2H), 1.91-1.95 (m, 1H), 1.63-1.59 (m, 1H), 1.47-1.42 (m, 2H), 1.38 - 1.31 (m, 1H); ESI: m/z: 292.1 (M+H)+.
[00329] Example 36: (±)-N-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexyl) pyrrolidine-2-carboxamide [1-36] .
Figure imgf000088_0001
(i)-Tert-butyl 2-(rel-(l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexylcarbamoyl)pyrrolidine- -carboxylate.
Figure imgf000088_0002
[00330] The title compound was synthesized in a similar fashion as Example 1 using (±)- l-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid and (±)-rel-(l S,3R)-3-(5-chloro-lH- indol-3-yl)cyclohexanamine. The crude product was purified by chromatography on silica gel to give (i)-tert-butyl 2-(rel-(l S,3R)-3-(5-chloro-lH-indol-3- yl)cyclohexylcarbamoyl)pyrrolidine-l-carboxylate. ESI: m/z: 346.1 (M+H)+.
(±)-N-(Rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl) cyclohexyl) pyrrolidine-2-carboxamide [I- 36].
Figure imgf000088_0003
[00331] A mixture of (±)-tert-butyl 2-(rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine- 1-carboxylate (0.34 mmol, 150 mg) in HCl/MeOH (15 mL, 3M) was heated to 70°C and then stirred for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H20 (10 mL) and the pH value was adjusted to 8.0 with Na2C03 (solid). The mixture was extracted with EtOAc (3 * 40 ml) and the combined organic fractions were dried over Na2S04 and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel to give (±)-N-(rel- (l S,3R)-3-(5-chloro-lH-indol-3-yl) cyclohexyl) pyrrolidine-2-carboxamide (1-36). ¾ MR (400 MHz, CD3OD) δ 7.45-7.44 (d, J = 2.0 Hz, 1H), 7.18-7.16 (d, J = 8.4 Hz, 1H), 6.95-6.91 (m, 2H), 3.80-3.72 (m, 1H), 3.55-3.51 (m, 1H), 2.96-2.90 (m, 1H), 2.84-2.78 (m, 2H), 2.14- 2.03 (m, 2H), 1.88-1.81 (m, 2H), 1.70-1.63 (m, 3H), 1.55-1.46 (m, 1H), 1.36-1.15 (m, 4H); ESI: m/z: 346.1 (M+H) +.
[00332] Example 37: (±)-N-(rel-(lS,3R)-3-(lH-indol-3-yl) cyclohexyl) pyrrolidine-2- carboxamide [1-37].
Figure imgf000089_0001
(±)-Benzyl 2-(rel-(lS,3R)-3-(5-chloro-lH-indol-3-yl)cyclohexylcarbamoyl)pyrrolidine-l- carboxylate.
Figure imgf000089_0002
[00333] The title compound was synthesized in a similar fashion as Example 1 using 1- (benzyloxycarbonyl)pyrrolidine-2-carboxylic acid and (±)-rel-(l S,3R)-3-(5-chloro-lH-indol- 3-yl)cyclohexanamine. The crude product was purified by chromatography on silica gel to give (±)-benzyl 2-(rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine- 1-carboxylate; ESI: m/z: 480.1 (M+H)+.
(±)-N-(Rel-(lS,3R)-3-(lH-indol-3-yl) cyclohexyl) pyrrolidine-2-carboxamide [1-37].
Figure imgf000089_0003
[00334] A mixture of (±)-benzyl 2-(rel-(l S,3R)-3-(5-chloro-lH-indol-3-yl) cyclohexylcarbamoyl) pyrrolidine- 1-carboxylate (0.31 mmol, 150 mg) and Pd/C (30 mg) in EtOH (10 mL) under H2 (1 atm) was stirred over 1 h at RT. The mixture was filtered through a pad of Celite and the filtrate was concentrated. The residue was purified by chromatography on silica gel to give (±)-N-(rel-(l S,3R)-3-(lH-indol-3-yl) cyclohexyl) pyrrolidine-2- carboxamide (1-37). ¾ NMR (400 MHz, CDCb) δ 7.94 (s, 1H), 7.68-7.65 (m, 1H), 7.54- 7.49 (m, 1H), 7.36-7.34 (d, J = 8.8 Hz, 1H), 7.20-7.16 (m, 1H), 7.13-7.08(m, 1H), 6.96- 6.95(m, 1H), 3.99-3.92 (m, 1H), 3.74-3.70 (m, 1H), 3.03-2.80 (m, 3H), 2.36-2.28 (m, 1H), 2.22-2.02 (m, 3H), 1.94-1.84 (m, 2H), 1.73-1.66 (m, 2H), 1.51-1.34(m, 4H), 1.22-1.13 (m, 1H); ESI: m/z: 342.1 (M+H)+.
[00335] Example 38 and 39: (S)-N-(rel-(lS,3R)-3-(5-chloro-2-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-38] and (S)-N-(rel-(lS,3S)-3-(5-chloro- 2-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-39].
Figure imgf000090_0001
1-38 1-39
[00336] The title compounds was synthesized in a similar fashion as Example 1 using (±)- 3-(5-chloro-2-methyl-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2-carboxylic acid. The crude product was purified by preparative HPLC using a Xbridge OBD column eluting with 60%-65% acetonitrile in water (10 mM H4HCO3) to obtain (S)-N-(rel-(l S,3R)- 3-(5-chloro-2-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-38); ¾ NMR (400 MHz, CDCb) δ 7.80 (br s, 1H), 7.58 (d, J = 2.0 Hz, 1H), 7.16 (d, J = 8.4 Hz, 1H), 7.03 (dd, J = 8.4, 2.0 Hz, 1H), 6.62 (d, J = 8.8 Hz, 1H), 4.31-4.35 (m, 1H), 3.82-4.00 (m, 3H), 2.80-2.88 (m, 1H), 2.36 (s, 3H), 2.20-2.30 (m, 1H), 1.91-2.08 (m, 4H), 1.67-1.89 (m, 5H), 1.48-1.57 (m, 1H), 1.24-1.34 (m, 1H); ESI: m/z: 361.1 (M+H)+ and (S)-N-(rel-(l S,3S)- 3-(5-chloro-2-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-39); ¾ NMR (500 MHz, CDCb) δ 7.77 (br s, 1H), 7.51-7.58 (m, 1H), 7.01-7.05 (m, 1H), 6.61-7.17 (m, 2H), 3.85-4.44 (m, 4H), 2.80-2.90 (m, 1H), 2.36-2.38 (m, 3H), 2.24-2.35 (m, 1H), 1.22- 2.16 (m, 11H); ESI: m/z: 361.1 (M+H)+.
[00337] Example 40 and 41: (S)-N-(rel-(lS,3R)-3-(5-methoxy-7-methyl-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide [1-40] and (S)-N-(rel-(lS,3S)-3-(5-methoxy- 7-methyl-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide [1-41].
Figure imgf000091_0001
[00338] The title compounds were synthesized in a similar fashion as Example 1 using (±)-3-(5-methoxy-7-methyl-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2- carboxylic acid. The crude product was purified by prep-HPLC using Xbridge OBD column eluting with 45-67% acetonitrile in water (10 mM H4HCO3) to give (S)-N-(rel-(l S,3R)-3- (5-methoxy-7-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-40); ¾ NMR (400 MHz, CD3OD) δ 6.97 (s, 1H), 6.89 (d, J = 2.0 Hz, 1H), 6.57 (s, 1H), 4.29-4.25 (m, 1H), 4.03-3.83 (m, 3H), 3.81 (s, 3H), 2.90(t, J= 11.6 Hz, 1H), 2.42 (s, 3H), 2.31-2.20 (m, 2H), 2.07-2.02 (m, 1H), 1.98-1.85 (m, 5H), 1.66-1.39 (m, 4H); (ESI): m/z: 357.1 (M+H)+, and (S)-N-(rel-(l S, 3S)-3-(5-methoxy-7-methyl-lH-indol-3-yl) cyclohexyl) tetrahydrofuran- 2-carboxamide (1-41); ¾ NMR (400 MHz, CD3OD) δ 7.05 (d, J= 3.2 Hz, 1H), 6.84-6.80 (m, 1H), 6.58 (s, 1H), 4.37-4.33 (m, 1H), 4.21-4.20 (m, 1H), 4.03-4.01 (m, 1H), 3.95-3.89 (m, 1H), 3.79 (d, J = 2.0 Hz, 3H), 3.15-3.07 (m, 1H), 2.43 (s, 3H), 2.34-2.26 (m, 1H), 2.11-1.83 (m, 6H), 1.72-1.68 (m, 5H); ESI: m/z: 357.1 (M+H)+.
[00339] Example 42: (S)-N-rel-((lS,3R)-3-(6-(benzyloxy)-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carbox .
Figure imgf000091_0002
[00340] The title compound was synthesized in a similar fashion as Example 1 using 3-(6- (benzyloxy)-5-methoxy-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2- carboxylic acid. The crude product was purified by chromatography on silica gel to obtain (S)-N-rel-((l S,3R)-3-(6-(benzyloxy)-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran- 2-carboxamide (1-42); ¾ NMR (400 MHz, CDCI3) δ 7.76 (s, 1H), 7.46 (d, J = 7.6 Hz, 2H), 7.36 (t, J = 7.6, 14.8 Hz, 2H) , 7.29 (d, J = 7.6 Hz, 1H), 7.06 (d, J = 5.2 Hz, 1H), 6.85-6.78 (m, 2H), 7.56 (d, J = 8.4 Hz, 1H), 5.17 (s, 2H), 4.35-4.31 (m, 1H), 4.03-3.81 (m, 6H), 2.94- 2.88 (m, 1H), 2.36-2.24 (m, 2H), 2.10-2.00 (m, 3H), 1.94-1.79(m, 3H), 1.61-1.54 (m, 1H), 1.46-1.16 (m, 3H); ESI: m/z: 449.1 (M+H)+. [00341] Example 43: (S)-N-rel-((lS,3R)-3-(6-hydroxy-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide [1-43].
Figure imgf000092_0001
[00342] A mixture of (S)-N-rel-((l S,3R)-3-(6-(benzyloxy)-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide (1.16 mmol, 500 mg), Pd/C (100 mg) in CH3OH (20 mL) under H2 was stirred over 1 h at RT. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified by chromatography (silica gel, PE/EtOAc=10/6) to obtain (S)-N-rel-((l S,3R)-3-(6-hydroxy-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide (1-43). ¾ MR (400 MHz, CDCb) δ 7.80 (s, 1H), 7.00 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 1.6 Hz, 1H), 6.81-6.78 (m, 1H), 6.58 (d, J = 7.6 Hz, 1H), 5.74 (d, J = 2.4 Hz, 1H), 4.36-4.31 (m, 1H), 4.04-3.94 (m, 4H), 3.92-3.84 (m, 2H), 2.94-2.87 (m, 1H), 2.38-2.25 (m, 2H), 2.10-2.04 (m, 2H), 1.95-1.79 (m, 3H), 1.62-1.54 (m, 1H), 1.43-1.28 (m, 2H), 1.22-1.13 (m, 1H); ESI: m/z: 359.1 (M+H)+.
[00343] Example 44 and 45: (S)-N-((lR,3S)-3-(6-(2-amino-2-oxoethoxy)-5-methoxy- lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide and (S)-N-((l S,3R)-3-(6-(2- amino-2-oxoethoxy)-5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide [1-44] an -45].
Figure imgf000092_0002
[00344] To a solution of (S)-N-(rel-(l S,3R)-3-(6-hydroxy-5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (50 mg, 0.14 mmol) and 2-chloroacetamide (26 mg, 0.28 mmol) in DMF (2 mL), was added potassium carbonate (58 mg, 0.42 mmol). The mixture was stirred at 60 °C for 12 h. The reaction mixture was treated with water (10 mL) and extracted with EtOAc (3 * 20 mL). The combined organic fractions were dried with Na2SC"4, filtered and concentrated. The residue was purified by preparative HPLC using a PHLEX ODS column eluting with 45%-75% acetonitrile in water (0.01% H3 + 10 mM H4HC03) to give (S)-N-((lR,3S)-3-(6-(2-amino-2-oxoethoxy)-5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (S)-N-((l S,3R)-3-(6-(2-amino-2- oxoethoxy)-5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-44), (retention time 6.70 min); ¾ MR (400 MHz, OMSO-d6) δ 10.52 (brs, 1H), 7.55 (d, J = 8.0 Hz, 1H), 7.43 (brs, 1H), 7.38 (brs, 1H), 7.06 (s, 1 H), 6.91-6.90 (m, 2H), 4.39 (s, 2H), 4.18- 4.15 (m, 1H), 3.89 (dd, J = 6.4, 14.4 Hz, 1H), 3.80 (s, 3H), 3.79-3.71 (m, 2H), 2.84-2.78 (m, 1H), 2.12-1.75 (m, 8H), 1.54-1.45 (m, 2H), 1.34-1.25 (m, 2H); ESI: m/z: 416.2 (M+H)+; and (S)-N-((lR,3S)-3-(6-(2-amino-2-oxoethoxy)-5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (S)-N-((l S,3R)-3-(6-(2-amino-2- oxoethoxy)-5-methoxy-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-45), (retention time 6.56 min) as a white solid; ¾ MR (400 MHz, DMSO-<f6) δ 10.52 (brs, 1H), 7.54 (d, J = 8.4 Hz, 1H), 7.44 (brs, 1H), 7.38 (brs, 1H), 7.06 (s, 1H), 6.91-6.90 (m, 2H), 4.39 (s, 2H), 4.19-4.16 (m, 1H), 3.89-3.70 (m, 6H), 2.84-2.78 (m, 1H), 2.10-1.73 (m, 8H), 1.52- 1.43 (m, 2H), 1.35-1.26 (m, 2H); ESI: m/z: 416.2 (M+H)+.
[00345] Example 46 and 47: (S)-N-(rel-(3R,5R)-5-(5-chloro-lH-indol-3-yl) tetrahydro-2H-pyran-3-yl) tetrahydrofuran-2-carboxamide [1-46] and (S)-N-(rel- (3R,5S)-5-(5-chloro-lH-indol-3-yl) tetrahydro-2H-pyran-3-yl) tetrahydrofuran-2- carboxamide [1-47].
Figure imgf000093_0001
1-46 1-47
[00346] The title compounds were synthesized in a similar fashion as Example 1 using of 5-(5-chloro-lH-indol-3-yl) tetrahydro-2H-pyran-3 -amine and (S)-tetrahydrofuran-2- carboxylic acid. The crude product was purified by preparative HPLC using a PHLEX ODS column eluting with 34%-64% acetonitrile in water (0.1% H4OH) to give (S)-N-(rel- (3R,5R)-5-(5-chloro-lH-indol-3-yl)tetrahydro-2H-pyran-3-yl)tetrahydrofuran-2-carboxamide (1-46); ¾ MR(500 MHz, CD3OD) δ 7.60-7.59 (m, 1H), 7.31 (d, J = 10.0 Hz, 1H), 7.16 (d, J= 9.0 Hz, 1H), 7.06 (d, J= 10.5 Hz, 1H), 4.28-4.25 (m, 1H), 4.12-4.3.79 (m, 5H), 3.47-3.35 (m, 1H), 3.29-3.19 (m, 2H), 2.28-2.21 (m, 2H), 1.97-1.79 (m, 4H); ESI: m/z: 349.1 (M+H)+ and (S)-N-(rel-(3R,5S)-5-(5-chloro-lH-indol-3-yl)tetrahydro-2H-pyran-3-yl)tetrahydrofuran- 2-carboxamide (1-47); ¾ NMR (500 MHz, CD3OD) δ 7.45-7.40 (m, 1H), 7.21-7.19 (m, 1H), 7.08 (s, 1H), 6.96-6.94 (m, 1H), 4.28-4.23 (m, 1H), 3.98-3.94 (m, 3H), 3.84-4.81 (m, 1H), 3.66-3.64 (m, 2H), 3.49-3.42 (m, 1H), 3.27-3.22 (m, 1H), 2.23-2.16 (m, 1H), 2.09-2.04 (m, 1H), 2.01-1.94 (m, 1H), 1.91-1.83 (m, 3H); ESI: m/z: 349.1 (M+H) +. [00347] Example 48 and 49: (S)-N-((lS,3S)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide and (S)-N-(lR,3R)-3-(5-methyl-lH-indol- 3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-48] and [1-49].
Figure imgf000094_0001
[00348] The title compound was synthesized in a similar fashion as Example 1 using (±)- 3-(5-methyl-lH-indol-3-yl)cyclohexanamine and (S)-tetrahydrofuran-2-carboxylic acid. The product was purified by preparative HPLC using a PHLEX ODS column eluting with 34%- 64% acetonitrile in water (0.1% H4OH) to give (±)-(2S)-N-(3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide. ESI: m/z: 327.2 (M+H)+ . The individual trans diastereomers were isolated by chiral SFC using a OJ-H column eluting with 35% MeOH (0.1% H4OH) in carbon dioxide to give (S)-N-((l S,3S)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (S)-N-((lR,3R)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-48), (ee 99.17%), retention time 3.81 min);. ¾ NMR (500 MHz, CD3OD): δ 7.33 (s, 1H), 7.21 (d, J = 8.0 Hz, 1H), 7.05 (s, 1H), 6.92 (d, J =8.5 Hz, 1H), 4.38-4.35 (m, 1H), 4.22-4.20 (m, 1H), 4.09-4.05 (m, 1H), 3.96-3.92 (m, 1H), 3.18-3.15 (m, 1H), 2.42 (s, 3H), 2.35-2.26 (m, 1H), 2.10-1.94 (m, 6H), 1.72-1.64 (m, 5H); ESI: m/z: 327.2 (M+H)+; and (S)-N-((lR,3R)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (S)-N-((l S,3S)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-49), (ee 96.31%, retention time 2.60 min); ¾ MR (500 MHz, MeOO-d4): δ 7.94 (s, 1H), 7.21 (d, J = 8.5 Hz, 1H), 7.03 (s, 1H), 6.92 (d, J =8.0 Hz, 1H), 4.38-4.35 (m, 1H), 4.22-4.20 (m, 1H), 4.10-4.08 (m, 1H), 3.95-3.93 (m, 1H), 3.13-3.10 (m, 1H), 2.42 (s, 3H), 2.35-2.29 (m, 1H), 2.13-1.97 (m, 5H), 1.96-1.94 (m, 1H), 1.74-1.70 (m, 5H); ESI: m/z: 327.2 (M+H)+.
[00349] Example 50 and 51: (S)-N-((lS,3R)-3-(5-methyl-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide and (S)-N-((lR,3S)-3-(5-methyl-lH-indol- 3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-50] and [1-51].
Figure imgf000094_0002
[00350] (±)-(2S)-N-(3-(5-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide from Examples 47 and 49 was separated into the individual cis diastereomers by chiral SFC using a AD-H column eluting with 35% MeOH (0.1% H4OH) in carbon dioxide to give (S)-N-((l S,3R)-3-(5-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide or (S)-N-((lR,3S)-3-(5-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2- carboxamide (1-50), (ee 99.17%, retention time 3.30 min); ¾ MR (500 MHz, CD3OD): δ 7.25 (s, 1H), 7.09 (d, J = 8.5 Hz, 1H), 6.84 (s, 1H), 6.80 (d, J =8.5 Hz, 1H), 4.17-4.14 (m, 1H), 3.91-3.72 (m, 3H), 2.83-2.79 (m, 1H), 2.30 (s, 3H), 2.17-2.09 (m, 2H), 1.96-1.93 (m, 1H), 1.86-1.76 (m, 5H), 1.51-1.12 (m, 4H); ESI: m/z: 327.2 (M+H)+; and (S)-N-((lR,3S)-3- (5-methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (S)-N-((l S,3R)-3-(5- methyl-lH-indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-51), (ee 99.61%), retention time 2.43 min): ¾ NMR (500 MHz, CD3OD): δ 7.38 (s, 1H), 7.20 (d, J = 8.0 Hz, 1H), 6.96 (s, 1H), 6.92 (d, J = 8.5 Hz, 1H), 4.30-4.27 (m, 1H), 4.00-3.83 (m, 3H), 2.97-2.91 (m, 1H), 2.43 (s, 3H), 2.30-2.17 (m, 2H), 2.07-2.05 (m, 1H), 2.03-1.86 (m, 5H), 1.66-1.24 (m, 4H); ESI 327.2 (M+H)+.
[00351] Examples 52 and 53: (R)-N-((lS,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide and (R)-N-((lR,3S)-3-(5-methoxy-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-52] and [1-53].
Figure imgf000095_0001
[00352] The title compounds were synthesized in a similar fashion as Example 1 using (±)-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine and (R)-tetrahydrofuran-2-carboxylic acid. The cis diastereomers were separated by chiral SFC using a AS-H column eluting with 25% MeOH(0.1%NH4OH) in carbon dioxide to give (R)-N-((l S,3R)-3-(5-methoxy-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (R)-N-((1R,3 S)-3-(5-methoxy-lH- indol-3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-52), (ee 94.8%>, retention time 4.74 min); ¾ NMR (400 MHz, CD3OD) δ 7.22 (d, J = 8.8 Hz, 1 H), 7.07 (d, J = 2.8 Hz, 1 H), 6.99 (s, 1 H), 6.76 (dd, J = 2.4, 9.6 Hz, 1 H), 4.28 (dd, J = 5.6, 8.4 Hz, 1 H), 4.01-3.84 (m, 6 H), 2.97-2.89 (m, 1 H), 2.31-2.19 (m, 2 H), 2.08-2.05 (m, 1 H), 1.98-1.84 (m, 5 H), 1.64-1.34 (m, 4 H); ESI: m/z: 343.2 (M+H)+; and (R)-N-((lR,3S)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide or (R)-N-((l S,3R)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-53), (ee 99.1%, retention time 5.12 min); ¾ MR (400 MHz, CD3OD) δ 7.22 (d, J= 7.2 Hz, 1 H), 7.07 (d, J= 2.0 Hz, 1 H), 6.96 (s, 1 H), 6.76 (dd, J = 2.4, 7.2 Hz, 1 H), 4.27 (dd, J = 4.4, 6.4 Hz, 1 H), 4.01-3.82 (m, 6 H), 2.92-2.86 (m, 1 H), 2.29-2.20 (m, 2 H), 2.05-2.03 (m, 1 H), 1.98-1.85 (m, 5 H), 1.63-1.29 (m, 4 H); ESI: m/z: 343.2 (M+H)+.
[00353] Examples: 54 and 55: (S)-N-(rel-(3R,5R)-5-(5-methoxy-lH-indol-3- yl)piperidin-3-yl)tetrahydrofuran-2-carboxamide [1-54] and (S)-N-(rel-(3R,5S)-5-(5- methoxy-lH-indol-3-yl)piperidin-3-yl)tetrahydrofuran-2-carboxamide [1-55].
Schem
Figure imgf000096_0001
1-54 1-55
(±)-Tert-butyl 3-amino-5-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxylate.
Figure imgf000096_0002
[00354] A mixture of (±)-tert-butyl 3-(5-methoxy-lH-indol-3-yl)-5-oxopiperidine-l- carboxylate (0.52 mmol, 180 mg), NH4OAc (15.7 mmol, 1.21 g) and NaBH3CN (1.3 mmol, 82 mg) in CH3OH (20 mL) was stirred for 1 h at RT. The reaction was poured into water and extracted with EtOAc (3 * 50 mL) and concentrated under reduced pressure to obtain 70 mg of (i)-tert-butyl 3-amino-5-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxylate.
(±)-(S)-Tert-butyl 3-(tetrahydrofuran-2-carboxamide)-5-(5-methoxy-lH-indol-3- yl)piperidine-l-carboxylate.
Figure imgf000097_0001
[00355] A mixture of (±)-tert-butyl 3-amino-5-(5-methoxy-lH-indol-3-yl)piperidine-l- carboxylate (0.20 mmol , 70 mg), (S)-tetrahydrofuran-2-carboxylic acid (0.41 mmol, 48 mg) , HATU (0.27 mmol, 126 mg) and DIEA (0.41 mmol, 55 mg) in DMF(5 mL) was stirred for 2 h at RT. The reaction was poured into water and extracted with EtOAc (3 * 50 mL). The combined organic fractions were dried over Na2S04 and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel to obtain (±)-(S)- tert-butyl-3 -(tetrahydrofuran-2-carboxamide)-5 -(5 -methoxy- lH-indol-3 -yl)piperidine- 1 - carboxylate. ESI: m/z: 388.2 (M+H)+.
(S)-N-(rel-(3R,5R)-5-(5-methoxy-lH-indol-3-yl)piperidin-3-yl)tetrahydrofuran-2- carboxamide [1-54] and (S)-N-(rel-(3R,5S)-5-(5-methoxy-lH-indol-3-yl)piperidin-3- yl)tetrahydrofuran-2-carboxamide [1-55] .
Figure imgf000097_0002
1-54 1-55
[00356] A mixture of (±)-(S)-tert-butyl-3-(tetrahydrofuran-2-carboxamide)-5-(5-methoxy- lH-indol-3-yl)piperidine-l-carboxylate (100 mg, 0.23 mmol), TFA (258 mg, 2.36 mmol) in DCM (1.0 mL) was stirred at rt overnight. The resulting mixture was concentrated in vacuo and purified by prep-HPLC using Xbridge OBD column eluting with 45-67% acetonitrile in water (10 mM NH4HCO3) to give (S)-N-(rel-(3R,5R)-5-(5-methoxy-lH-indol-3-yl)piperidin- 3-yl)tetrahydrofuran-2-carboxamide (1-54); ¾ NMR (400 MHz, OMSO-d6) δ 10.68 (br s, 1H), 7.53-7.52 (m, 1H), 7.23 (d, J= 8.8 Hz, 1H), 7.03 (s, 2H), 6.72 (d, J= 8.8 Hz, 1H), 4.20- 4.17 (m, 1H), 3.91-3.89 (m, 1H), 3.80-3.73 (m, 5H), 3.42 (br s, 2H), 3.11-3.09 (m, 1H), 2.94- 2.92 (m, 2H), 2.38-2.33 (m, 1H), 2.13-2.04 (m, 2H), 1.87-1.69 (m, 4H); ESI: m/z: 344.7 (M+H)+; and (S)-N-(rel-(3R,5S)-5-(5-methoxy-lH-indol-3-yl)piperidin-3-yl)tetrahydrofuran- 2-carboxamide (1-55). ¾ NMR (400 MHz, DMSO- d6) δ 10.71 (br s, 1H), 7.56 (br s, 1H), 7.23 (d, J = 8.6 Hz, 1H), 7.09-6.99 (m, 2H), 6.72 (d, J = 8.7 Hz, 1H), 4.21-4.18 (m, 1H), 3.90-3.84 (m, 5H), 3.76-3.71 (m, 4H), 3.09 (br s, 1H), 2.96-2.90 (m, 1H), 2.40 (br s, 1H), 2.14-2.02 (m, 2H), 1.91-1.70 (m, 4H); ESI 344.7 (M+H)+.
[00357] Example 56: (S)-N-(rel-(lS,3R)-3-(6-(2-hydroxyethyl)-5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-56].
Scheme 3
Figure imgf000098_0001
Tert-butyl 6-(benzyloxy)-5-methoxy-3-(rel-(lS,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-lH-indole- -carboxylate.
Figure imgf000098_0002
[00358] A mixture of (±)-(S)-N-rel-((l S,3R)-3-(6-(benzyloxy)-5-methoxy-lH-indol-3-yl) cyclohexyl) tetrahydrofuran-2-carboxamide (1.25 g, 2.79 mmol), B0C2O (1.83 g, 8.37 mmol), TEA (0.85 g, 8.37 mmol) and DMAP (0.18 g, 0.84 mmol) in DCM (20 mL) was stirred for 2 h at RT. The reaction was poured into water and extracted with EtOAc (3 * 50 mL). The organic fractions were combined and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, PE/EtOAc=10/6) to obtain tert-butyl 6-(benzyloxy)-5- methoxy-3-(rel-(l S,3R)-3-((S)-tetrahydrofuran-2-carboxamido) cyclohexyl) carboxylate. ESI: m/z: 549.1 (M+H)+.
Tert-butyl 6-hydroxy-5-methoxy-3-(rel-(lS,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-lH-indole- -carboxylate.
Figure imgf000099_0001
[00359] A mixture of tert-butyl 6-(benzyloxy)-5-methoxy-3-(rel-(l S,3R)-3-((S)- tetrahydrofuran-2-carboxamido)cyclohexyl)-lH-indole-l-carboxylate (1.5 g, 2.74 mmol), Pd/C (0.4 g) in CH3OH (15 mL) under H2 was stirred over 1 h at RT. The mixture was filtered and concentrated and the residue was purified by chromatography (silica gel, PE/EtOAc=10/6) to obtain 980 mg of tert-butyl 6-hydroxy-5-methoxy-3-(rel-(l S,3R)-3-((S)- tetrahydrofuran-2-carboxamido)cyclohexyl)-lH-indole-l-carboxylate. ESI: m/z: 459.1 (M+H)+.
Tert-butyl 5-methoxy-3-(rel-(l S,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-6-(trifluoromethylsulfonyloxy)-lH-indole-l-carboxylate.
Figure imgf000099_0002
[00360] To a solution of tert-butyl 6-hydroxy-5-methoxy-3-(rel-(l S,3R)-3-((S)- tetrahydrofuran-2-carboxamido)cyclohexyl)-lH-indole-l-carboxylate (980 mg, 2.14 mmol) and TEA (432 mg, 4.28 mmol) in dry DCM (40 mL) was added drop wise trifluoromethanesulfonic anhydride (905 mg, 3.21 mmol) at -40 °C. The mixture was stirred for 2 h at -40 °C. The reaction was quenched with MeOH (2 mL) and the mixture was concentrated at 30 °C. The residue was purified by chromatography (silica gel, PE/EtOAc=10/5) to obtain 600 mg of tert-butyl 5-methoxy-3-(rel-(l S,3R)-3-((S)- tetrahydrofuran-2-carboxamido)cyclohexyl)-6-(trifluoromethylsulfonyloxy)- lH-indole- 1 - carboxylate. ESI: m/z 591.1 (M+H)+.
Tert-butyl 5-methoxy-3-(rel-(l S,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-6-vinyl-lH-indole-l-carboxylate.
Figure imgf000100_0001
[00361] To a solution of tert-butyl 5-methoxy-3-(rel-(l S,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-6-(trifluoromethylsulfonyloxy)-lH-indole-l-carboxylate (200 mg, 0.34 mmol), tributyl(vinyl)stannane (216 mg, 0.68 mmol) in DMF (5 mL), were added lithium chloride (72 mg, 1.7 mmol) and Pd(dppf)Cl2 (50 mg, 0.068 mmol). The mixture was stirred at 100 °C for 16h under N2 atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3 * 30 mL). The combined organic fractions were dried with Na2S04, filtered and concentrated. The residue was diluted with DCM (5 mL). To this solution was added Boc20 (87 mg, 0.4 mmol), triethylamine (61 mg, 0.6 mmol) and N,N- Dimethyl-4-aminopyridine (5 mg, 0.04 mmol). The mixture was stirred at room temperature for 3h. The mixture was concentrated and the residue was purified by flash chromatography (silica gel, PE/EtOAc=10/l) to give tert-butyl 5-methoxy-3-(rel-(l S,3R)-3-((S)- tetrahydrofuran-2-carboxamido)cyclohexyl)-6-vinyl-lH-indole-l-carboxylate. ESI: m/z: 469.1 (M+H)+.
Tert-butyl 6-(2-hydroxyethyl)-5-methoxy-3-(rel-(lS,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-lH-indole- -carboxylate.
Figure imgf000100_0002
[00362] To a solution of tert-butyl 5-methoxy-3-(rel-(l S,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-6-vinyl-lH-indole-l-carboxylate (50 mg, 0.11 mmol) in THF (5 mL), was added borane tetrahydrofuran complex (1 M in THF, 0.33 mL, 0.33 mmol) at -78 °C. the mixture was warmed to 0 °C and stirred at this temperature for 3h. The mixture was diluted with 10% NaOH (5 mL) and 30% hydrogen peroxide (1 mL) was added drop wise. The mixture was stirred at room temperature for 3h and then extracted with EtOAc (3 * 20 mL). The combined organic layers were dried with Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, DCM/MeOH=10/l) to give tert- butyl 6-(2-hydroxyethyl)-5-methoxy-3-(rel-(l S,3R)-3-((S)-tetrahydrofuran-2- carboxamido)cyclohexyl)-lH-indole-l-carboxylate. ESI: m/z: 487.1 (M+H)+.
(S)-N-(Rel-(lS,3R)-3-(6-(2-hydroxyethyl)-5-methoxy-lH-indol-3- yl)cyclohexyl)tetrahydrofuran-2-carboxamide [1-56].
Figure imgf000101_0001
[00363] To a solution of tert-butyl 6-(2-hydroxyethyl)-5-methoxy-3-((l S,3R)-3-((S)- tetrahydrofuran-2-carboxamido)cyclohexyl)-lH-indole-l-carboxylate (30 mg, 0.062 mmol) in DCM (1 mL), was added trifluoroacetic acid (1 mL). The mixture was stirred at room temperature for 6h. The pH of the mixture was was made neutral with sat. aqueous NaHCCb and then the mixture was concentrated under reduce pressure. The residue was purified by preparative HPLC using a Xbridge ODS column eluting with 30%-60% acetonitrile in water (10 mM H4HCO3) to give (S)-N-(rel-(l S,3R)-3-(6-(2-hydroxyethyl)-5-methoxy-lH-indol- 3-yl)cyclohexyl)tetrahydrofuran-2-carboxamide (1-56). ¾ MR (400 MHz, DMSO-<f6) δ 10.47 (brs, 1 H), 7.54 (t, J= 6.0 Hz, 1 H), 7.09 (s, 1 H), 6.98 (s, 1 H), 6.94 (d, , J = 2.0 Hz, 1 H), 4.56 (t, J = 4.0 Hz, 1 H), 4.19-4.15 (m , 1 H), 3.90-3.85 (m, 1 H), 3.79-3.71 (m, 5 H), 3.54 (m, 2 H), 2.84-2.75 (m, 2 H), 2.10-1.74 (m, 8 H), 1.51-1.45 (m, 2 H), 1.36-1.31 (m, 2 H); ESI: m/z: 387.1 (M+H)+.
[00364] Example 57: (±)-rel-N-((lS,3S)-3-(5-methoxy-lH-indol-3- yl)cyclohexyl)acetamide [1-57].
Scheme 4
Figure imgf000101_0002
(±)-Rel-(lS,3S)-N-Benzyl-3-(5-methoxy-lH-indol-3-yl)cyclohexanamine.
Figure imgf000102_0001
[00365] To a solution of (±)-3-(5-methoxy-lH-indol-3-yl)cyclohexanone (12.3 mmol, 3 g), phenylmethanamine (12.3 mmol, 1.3 g) in DCM (20 mL) was added NaBH4 (37 mmol, 1.4 g). The mixture was stirred at 30°C for 16h. The reaction mixture was treated with water (20 mL) and extracted with DCM (2 * 50 mL). The combined organic layers were dried with Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, PE/EtOAc=3/l) to give (±)-rel-(l S,3S)-N-Benzyl-3-(5-methoxy-lH-indol-3- yl)cyclohexanamine. ¾ MR (500 MHz, MeOD) δ 7.28-7.21 (m, 6H), 7.09 (d,lH), 6.88 (s, 1H), 6.77 (dd, 1H), 4.88 (s, 2H), 3.78 (s, 3H), 3.26(m, 1H, Ha), 2.89(m, 1H, Hb), 1.99- 1.57(m, 8H); ESI: m/z: 335.2 (M+H)+.
(±)-Rel-(lS,3S)-3-(5-Methoxy-lH-indol-3-yl)cyclohexanamine.
Figure imgf000102_0002
[00366] To a solution of (±)-rel-(l S,3S)-N-Benzyl-3-(5-methoxy-lH-indol-3- yl)cyclohexanamine (4.8 mmol, 1.6 g) in MeOH (20 mL) was added Pd(OH)2 (160 mg). The reaction mixture was stirred at 25 °C for 5h under H2 atmosphere. The reaction mixture was filtered and the filtrate was concentrated to give (±)-rel-(l S,3S)-3-(5-Methoxy-lH-indol-3- yl)cyclohexanamine and used to the next step without further purification. ESI: m/z: 245.2 (M+H)+.
(±)-Rel-N-((lS,3S)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)acetamide [1-57].
Figure imgf000102_0003
[00367] To a solution of (±)-rel-(l S, 3S)-3-(5-Methoxy-lH-indol-3-yl)cyclohexanamine (100 mg, 0.41 mmol) in DCE (3 mL), was added acetic anhydride (63 mg, 0.61 mmol) and Et3N (83 mg, 0.82 mmol). The mixture was stirred at 25 °C for 15h. Then the reaction mixture was concentrated and the residue was purified by preparative HPLC using a Xbridge OBD column eluting with 45%-75% acetonitrile in water(0.1% H4OH) to get (±)-rel-N- ((l S,3S)-3-(5-methoxy-lH-indol-3-yl)cyclohexyl)acetamide (I-57).1H NMR (400 MHz, DMSO-i/e) δ 10.58 (s, 1H), 7.87-7.89 (d, 1H, J = 7.2 Hz), 7.19-7.22 (d, 1H, J = 8.8 Hz), 7.04 (d, 1H, J = 2.0Hz), 6.97 (d, 1H, J =2.4 Hz), 6.68-6.71 (q, 1H, J = 2.4 Hz), 4.04 (brs, 1H), 3.74 (s, 3H), 3.13 (brs, 1H), 1.87-1.91 (m, 5H), 1.69-1.72 (brs, 2H), 1.49-1.57 (m, 4H); ESI: m/z: 287.1 (M+H)+.
[00368] Examples 58 and 59: ((R)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((R)- tetrahydrofuran-2-yl)methanone and ((S)-3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)((R)-tetrahydrofuran-2-yl)methanone [1-58] and [1-59].
Sche
Figure imgf000103_0001
Tert-butyl 5-(5-methoxy-lH-indol-3- l)-3,4-dihydropyridine-l(2H)-carboxylate.
Figure imgf000103_0002
[00369] To a solution of 5-methoxy-lH-indole (1.18 g, 8.0 mmol), tert-butyl 3- oxopiperidine-l-carboxylate (4.78 g, 24.0 mmol) in MeOH (80 mL) was added sodium methoxide (2.59 g, 48.0 mmol). The reaction mixture was stirred at reflux for 48h. The pH value of the reaction mixture was adjusted to 6 using acetic acid. The mixture was concentrated. The residue was diluted with water and the mixture was extracted with EtOAc (3 * 100 mL). The combined organic layers were dried with Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, PE/EtOAc=4/l) to give the product. ESI: m/z: 329.2 (M+H)+.
(±)-Tert-butyl 3-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxylate.
Figure imgf000104_0001
[00370] To a solution of tert-butyl 5-(5-methoxy-lH-indol-3-yl)-3,4-dihydropyridine- l(2H)-carboxylate (300 mg, 0.91 mmol) in MeOH (5 mL), was added Pd/C (30 mg). The mixture was stirred at room temperature for 12h under H2 atmosphere. The mixture was filtered and concentrated. The residue was purified by flash chromatography (silica gel, PE/EtOAc=4/l) to give (±)-tert-butyl 3-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxylate. ESI: m/z: 231.3 (M+H-100)+.
(±)-5-Methoxy-3-(piperidin-3-yl)-lH-indole hydrochloride.
Figure imgf000104_0002
[00371] A solution of (±)-tert-butyl 3-(5-methoxy-lH-indol-3-yl)piperidine-l- carboxylate(300 mg, 0.9 mmol) in 3M HCl in MeOH was stirred at room temperature for 3h. The mixture was concentrated to give (±)-5-methoxy-3-(piperidin-3-yl)-lH-indole hydrochloride. ESI: m/z: 231.3 (M+H)+.
((R)-3-(5-Methoxy-lH-indol-3-yl)piperidin-l-yl)((R)-tetrahydrofuran-2-yl)methanone and ((S)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((R)-tetrahydrofuran-2- yl)methanone [1-58] and [1-59].
Figure imgf000104_0003
[00372] To a solution of (±)-5-methoxy-3-(piperidin-3-yl)-lH-indole hydrochloride (66 mg, 0.25 mmol), (R)-tetrahydrofuran-2-carboxylic acid (32 mg, 0.28 mmol) and HATU (144 mg 0.38 mmol) in DMF (1 mL), was added DIPEA (1.0 mL). The mixture was stirred at room temperature for 2 h. The mixture diluted with EtOAc (100 ml). The mixture was washed with water (3 χ 20 mL), dried (Na2S04) and concentrated. The residue was purified by preparative HPLC using a PHLEX ODS column eluting with 45%-70% acetonitrile in water (0.1% H4OH) to give ((R)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((R)- tetrahydrofuran-2-yl)methanone or ((S)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((R)- tetrahydrofuran-2-yl)methanone (1-58), (retention time 1.308 min.); ¾ NMR (500 MHz, CDCb) δ 8.08-8.02 (m, 1H), 7.31-7.27 (m, 1H), 7.26-7.13 (m, 1H), 6.99-6.98 (m, 1H), 6.88- 6.83 (m, 1H), 4.72-4.64 (m, 2H), 4.44-3.98 (m, 2H), 3.92-3.88 (m, 1H), 3.87 (s, 3H), 3.20- 2.61 (m, 3H), 2.39-1.78 (m, 8H); ESI: m/z: 329.2 (M+H)+; and ((S)-3-(5-methoxy-lH-indol- 3-yl)piperidin-l-yl)((R)-tetrahydrofuran-2-yl)methanone or ((R)-3-(5-methoxy-lH-indol-3- yl)piperidin-l-yl)((R)-tetrahydrofuran-2-yl)methanone (1-59), (retention time 1.296 min); ¾ NMR (500 MHz, CDCb) δ 8.18-8.08 (m, 1H), 7.29-7.23 (m, 1H), 7.14-7.03 (m, 1H), 7.01- 6.95 (m, 1H), 6.89-6.83 (m, 1H), 4.85-4.60 (m, 2H), 4.26-3.74 (m, 4H), 3.86 (s, 3H), 3.13- 2.94 (m, 2H), 2.75-2.30 (m, 1H), 2.30-1.97 (m, 3H), 1.94-1.77 (m, 3H), 1.66-1.62 (m, 1H); ESI: m/z: 329.2 (M+H)+.
[00373] Example 60: (±)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)methanone [1-60].
Figure imgf000105_0001
[00374] The title compound (1-60) was synthesized using cyclopentane carboxylic acid and (±)-5-methoxy-3-(piperidin-3-yl)-lH-indole hydrochloride in a similar fashion as examples 57 and 58. ¾ NMR (500 MHz, CDCb) δ 8.03-7.94 (m, 1H), 7.29-7.23 (m, 1H), 7.14-6.98 (m, 2H), 6.89-6.84 (m, 1H), 4.85-4.67 (m, 1H), 4.22-4.19 (m, 1H), 3.87 (s, 3H), 3.17-2.93 (m, 3H), 2.82-2.63 (m, 1H), 2.21-2.20 (m, 1H), 1.95-1.75 (m, 8H), 1.71-1.68 (m, 1H), 1.65-1.26 (m, 2H); ESI: m/z: 327.2 (M+H)+.
[00375] Example 61 and 62: (S)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)methanone and (R)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)methanone [1-61] and [1-62
Figure imgf000105_0002
[00376] (±)-Cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)methanone was purified by chiral SFC using an AD-H column eluting with 35% MeOH (0.1%NH4OH) in carbon dioxide to give (S)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)methanone or (R)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)methanone (I- 61), (ee 100%, retention time 3.45 min); ¾ NMR (500 MHz, CDCb) δ 8.08-7.98 (m, 1H), 7.35-7.23 (m, 1H), 7.14-6.97 (m, 2H), 6.89-6.83 (m, 1H), 4.89-4.64 (m, 1H), 4.22-3.96 (m, 1H), 3.86 (s, 3H), 3.17-2.61 (m, 4H), 2.23-2.20 (m, 1H), 2.03-1.43 (m, 11H), ESI: m/z: 327.2 (M+H)+; and (R)-cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)methanone or (S)- cyclopentyl(3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)methanon (1-62), (ee 100%, retention time 5.09 min); ¾ NMR (500 MHz, CDCb) δ 8.03-7.91 (m, 1H), 7.22-7.16 (m, 1H), 7.07- 6.90 (m, 2H), 6.82-6.77 (m, 1H), 4.78-4.60 (m, 1H), 4.14-3.89 (m, 1H), 3.80 (s, 3H), 3.10- 2.56 (m, 4H), 2.14-2.13 (m, 1H), 1.94-1.42 (m, 11H); ESI: m/z: 327.2 (M+H)+.
[00377] Example 63: (±)-N-cyclopentyl-3-(5-methoxy-lH-indol-3-yl)piperidine-l- carboxamide [1-63].
Figure imgf000106_0001
[00378] To a solution of (±)-5-methoxy-3-(piperidin-3-yl)-lH-indole hydrochloride (160 mg, 0.60 mmol) and isocyanatocyclopentane (68 mg, 0.61 mmol) in THF (10 mL), was added TEA (1.0 mL). The mixture was stirred at room temperature for 3 h. The mixture diluted with EtOAc (100 ml). The mixture was washed with water (3 x 20 mL) the organic fraction was dried (Na2S04) and concentrated. The residue was purified by preparative HPLC using a PHLEX ODS column eluting with 34%-64% acetonitrile in water (0.1%NH4OH) to give (±)-N-cyclopentyl-3-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxamide (1-63). ¾ NMR (500 MHz, CDCb) δ 8.12 (b, 1H), 7.26 (d, J = 11 Hz, 1H), 7.07 (d, J = 3.0 Hz, 1H), 6.99 (d, J = 2.5 Hz, 1H), 6.86 (dd, J = 11.0, 2.5 Hz, 1H), 4.37-4.36 (m, 1H), 4.15-4.08 (m, 1H), 4.04-3.94 (m, 2H), 3.86 (s, 3H), 3.05-2.90 (m, 3H), 2.17-1.58 (m, 10H), 1.39-1.25 (m, 2H); ESI: m/z: 342.3 (M+H)+.
[00379] Example 64 and 65: (R)-N-cyclopentyl-3-(5-methoxy-lH-indol-3- yl)piperidine-l-carboxamide and (S)-N-cyclopentyl-3-(5-methoxy-lH-indol-3- yl)piperidine-l-carboxami -64] and [1-65].
Figure imgf000106_0002
[00380] (±)-N-cyclopentyl-3-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxamide
(Example 62) was purified by chiral SFC using a AD-H column eluting with 35% MeOH(0.1% H4OH) in carbon dioxide to give (R)-N-cyclopentyl-3-(5-methoxy-lH-indol-3- yl)piperidine-l-carboxamide or (S)-N-cyclopentyl-3-(5-methoxy-lH-indol-3-yl)piperidine-l- carboxamide (1-64), (ee 98.4%, retention time 3.15 min); ¾ NMR (500 MHz, CDCb) δ 7.97 (b, 1H), 7.26 (d, J = 11.0 Hz, 1H), 7.08 (d, J = 2.5 Hz, 1H), 7.00 (d, J = 2.0 Hz, 1H), 6.86 (dd, J = 8.5, 2.0 Hz, 1H), 4.35-4.34 (m, 1H), 4.14-3.95 (m, 3H), 3.87 (s, 3H), 3.03-2.90 (m, 3H), 2.17-1.59 (m, 10H), 1.35-1.26 (m, 2H); ESI: m/z: 342.3 (M+H)+ and (S)-N-cyclopentyl- 3-(5-methoxy-lH-indol-3-yl)piperidine-l-carboxamide or (R)-N-cyclopentyl-3-(5-methoxy- lH-indol-3-yl)piperidine-l-carboxamide (1-65), (ee 92.5%, retention time 4.62 min); ¾ MR (500 MHz, CDCb) δ 7.95 (b, 1H), 7.26 (d, J = 11 Hz, 1H), 7.08 (d, J = 2.0 Hz, 1H), 7.00 (d, J = 2.0 Hz, 1H), 6.86 (dd, J = 9.0, 2.5 Hz, 1H), 4.35-4.33 (m, 1H), 4.14-3.95 (m, 3H), 3.87 (s, 3H), 3.03-2.90 (m, 3H), 2.18-1.58 (m, 10H), 1.35-1.25 (m, 2H); ESI: m/z: 342.3 (M+H)+.
[00381] Example 66 and 67: ((R)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((S)- tetrahydrofuran-2-yl)methanone and ((S)-3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)((S)-tetrahydrofuran-2-yl)methanone [1-66] and [1-67].
Figure imgf000107_0001
[00382] The title compound was synthesized in a similar fashion as Example 50 using (S)- tetrahydrofuran-2-carboxylic acid and (±)-5-methoxy-3-(piperidin-3-yl)-lH-indole hydrochloride. The mixture was purified by Prep-TLC eluting with PE/EtOAc=l/l to give ((R)-3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((S)-tetrahydrofuran-2-yl)methanone or ((S)- 3-(5-methoxy-lH-indol-3-yl)piperidin-l-yl)((S)-tetrahydrofuran-2-yl)methanone (1-66); ¾ NMR (400 MHz, CDCb) δ 8.18 (brs, 0.7 H), 8.11 (brs, 0.3 H), 7.30-7.23 (m, 1.7 H), 7.13 (d, J= 2.0 Hz, 0.3 H), 6.99-6.97 (m, 1 H), 6.88-6.83 (m, 1 H), 4.80-4.65 (m, 2 H), 4.44-4.41 (m, 0.7 H), 4.04-3.99 (m, 1.3 H), 3.92-3.86 (m, 4 H), 3.20-2.80 (m, 2.3 H), 2.68-2.61 (m, 0.7 H), 2.40-1.62 (m, 8 H); ESI: m/z: 329.2 (M+H)+; and ((S)-3-(5-methoxy-lH-indol-3-yl)piperidin- l-yl)((S)-tetrahydrofuran-2-yl)methanone or ((R)-3-(5-methoxy-lH-indol-3-yl)piperidin-l- yl)((S)-tetrahydrofuran-2-yl)methanone (1-67); ¾ NMR (400 MHz, CDCb) δ 8.07 (brs, 0.5H), 7.98 (brs, 0.5 H), 7.29-7.23 (m, 1H), 7.15-6.97 (m, 2H), 6.90-6.84 (m, 1H), 4.87-4.83 (m, 0.5H), 4.72-4.61 (m, 1.5H), 4.29-4.26 (m, 0.5H), 4.07-3.81 (m, 5.5H), 3.14-2.96 (m, 2H), 2.75-2.68 (m, 1H), 2.30-1.67 (m, 8H); ESI: m/z: 329.2 (M+H)+.
[00383] Example 68: (R)-N-(3-(5-methoxy-lH-indol-3-yl)phenyl)tetrahydrofuran-2- carboxamide [1-68].
Schem 6
Figure imgf000108_0001
Tert-butyl 5-methoxy-3-(3-nitrophenyl)-lH-indole-l-carboxylate.
Figure imgf000108_0002
[00384] The mixture of tert-butyl 3-bromo-5-methoxy-lH-indole-l-carboxylate (1.54 mmol, 500 mg), 3-nitrophenylboronic acid (1.54 mmol, 257 mg), Pd(PPh3)4 (0.15 mmol, 173 mg), Na2CO3(490 mg, 4.6 mmol), H20 (2mL) in dioxane (20mL) under an Argon atmosphere was stirred at 80 °C overnight. The crude mixture was purified by flash chromatography (silica gel, PE/EtOAc=20%) to give tert-butyl 5-methoxy-3-(3-nitrophenyl)-lH-indole-l- carboxylate. ESI: m/z: 369.1 (M+H)+.
Tert-butyl 3-(3-aminophenyl)-5-methoxy-lH-indole-l-carboxylate.
Figure imgf000108_0003
[00385] Hydrazine (0.2mL) was slowly added to a mixture of tert-butyl 5-methoxy-3-(3- nitrophenyl)-lH-indole-l-carboxylate (150 mg, 0.41 mmol), and Raney Ni (0.2mL) in CH3OH (20mL).The mixture was stirred at R.T. for 30 min. The mixture was filtered and concentrated to provide tert-butyl 3-(3-aminophenyl)-5-methoxy-lH-indole-l-carboxylate (130 mg). ESI: m/z: 339.2 (M+H)+.
Tert-butyl (R)- 5-methoxy-3-(3-(tetrahydrofuran-2-carboxamido)phenyl)-lH-indole-l- carboxylate.
Figure imgf000109_0001
[00386] A mixture of tert-butyl 3-(3-aminophenyl)-5-methoxy-lH-indole-l-carboxylate (70 mg, 0.21 mmol), (R)-tetrahydrofuran-2-carboxylic acid (26 mg, 0.23 mmol), HATU (120 mg, 0.31 mmol), TEA (64 mg, 0.63 mmol) in DMF (4 mL) was stirred at R.T. overnight. The mixture was diluted with water (20 mL) and extracted with EtOAc (2 * 50 mL). The combined organic fractions were dried with Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, PE/EtOAc=20%) to give tert-butyl (R)- 5- methoxy-3-(3-(tetrahydrofuran-2-carboxamido)phenyl)-lH-indole-l-carboxylate. ESI: m/z: 437.2 (M+H)+.
(R)-N-(3-(5-Methoxy-lH-indol-3-yl)phenyl)tetrahydrofuran-2-carboxamide [1-68].
Figure imgf000109_0002
[00387] A mixture of (R)-tert-butyl 5-methoxy-3-(3-(tetrahydrofuran-2- carboxamido)phenyl)-lH-indole-l-carboxylate (20 mg, 0.046 mmol), and TFA (5 mL) in DCM (5 mL) was stirred at R.T. for lh. The mixture was concentrated and the residue was purified by preparative HPLC using Xbridge OBD column eluting with 45%-75% acetonitrile in water (0.01% H3 +10 mM NH4HC03) to give (R)-N-(3-(5-methoxy-lH-indol-3- yl)phenyl)tetrahydrofuran-2-carboxamide (1-68). ¾ MR (400 MHz, CD3OD) δ 8.03 (s, 1H), 7.54-7.26 (m, 6H), 6.92- 6.73 (m, 1H), 4.47 (d, J = 2.2 Hz, 1H), 4.12 (s, 1H), 3.96 (d, J = 8.1 Hz, 1H), 3.86 (s, 3H), 2.43- 2.30 (m, 1H), 2.16-2.07 (m, 1H), 2.04-1.95 (m, 2H), ESI: m/z: 337.2 (M+H)+.
[00388] Example 69: (S)-N-(3-(5-methoxy-lH-indol-3-yl)phenyl)tetrahydrofuran-2- carboxamide [1-69].
Figure imgf000110_0001
[00389] The title compound (1-69) was synthesized in a similar fashion as Example 67 using (S)-tetrahydrofuran-2-carboxylic acid and tert-butyl 5-methoxy-3-(3-nitrophenyl)-lH- indole-l-carboxylate in the amidation step followed by Boc-deprotection using TFA. ¾ NMR (400 MHz, CD3OD) δ 8.03 (s, 1H), 7.49-7.25 (m, 6H), 6.83 (dd, J = 8.8, 2.4 Hz, 1H), 4.46 (dd, J = 8.2, 6.0 Hz, 1H), 4.13 (dd, J = 14.7, 6.6 Hz, 1H), 3.95 (dd, J = 14.9, 6.8 Hz, 1H), 3.86 (s, 3H), 2.44-2.30 (m, 1H), 2.18-2.06 (m, 1H), 1.98 (p, J = 7.0 Hz, 2H); ESI: m/z: 337.2 (M+H)+.
MTHFD2 activity assay
[00390] Human MTHFD2 (residues 36-350 in Uniport ID P13995) was expressed as an N- terminal His6 tagged protein and purified in E.coli using nickel capture followed by size- exclusion chromatography.
[00391] MTHFD2 dehydrogenase activity oxidizes methylene tetrahyrofolate to methenyl- tetrahydrofolate with the concomitant reduction of NAD to NADH. MTHFD2 dehydrogenase activity was followed by coupling the generation of NADH to dipahorase catalyzed oxidation of resazurin to fluorescent resorufin. Briefly, control (DMSO) or compound was incubated with 0.2nM MTHFD2 for 15 minutes in 10.5 μΐ assay buffer containing lOOmM Hepes, pH 8.0, 5mM MgCb, 5mM KH2PO4, 0.05% BSA, 0.01% Tween- 20 in a 384 well Griener, low volume, black plate. ΙΟμΙ of substrate buffer containing 70μΜ CH2-THF, ImM NAD+, ΙΟΟμΜ Resazurin and 0.42 mg/ml diaphorase was then added to initiate the reaction and the reaction plate was incubated for 30 minutes. 5μ1 of 250mM EDTA was then added to each well to stop the reaction and MTHFD2 activity was quantified by measuring resorufin fluorescence emission at 598nm following excitation at 525nm. Activity in positive control wells containing the complete reaction mixture and DMSO alone was set to 100% while activity in negative control wells with the complete reaction mix without MTHFD2 was set to 0%. 11 concentrations of each compound were tested and the fluorescent data at each concentration was normalized to the positive and negative controls and represented as % inhibition. Median inhibitory concentration (IC50) was calculated by fitting % inhibition at each concentration to a 4 parameter logistic fit equation using Graphpad Prizm.
[00392] Table 3 shows the activity of selected compounds of this invention in the MTHFD2 inhibition assay. The compound numbers correspond to the compound numbers in Tables 1 and 2. Compounds having an activity designated as "A" provided an IC50 < 10 μΜ; compounds having an activity designated as "B" provided an IC50 10 - 30 μΜ; compounds having an activity designated as "C" provided an IC50 30 - 50 μΜ; compounds having an activity designated as "D" provided an IC50 > 50 μΜ.
Table 3. Human MTHFD2 Activity
Figure imgf000111_0001
Compound Number Human MTHFD2 activity
1-25 D
1-26 B
1-27 B
1-28 D
1-29 D
1-30 D
1-31 D
1-32 D
1-33 C
1-34 D
1-35 B
1-36 A
1-37 C
1-38 D
1-39 C
1-40 A
1-41 D
1-42 C
1-43 D
1-44 D
1-45 D
1-46 A
1-47 D
1-48 D
1-49 D
1-50 A
1-51 D
1-52 A
1-53 C
1-54 A
1-55 D
1-56 B
1-57 D
1-58 D
1-59 D
1-60 D
1-61 D
1-62 D
1-63 B
1-64 C
1-65 B Compound Number Human MTHFD2 activity
1-66 B
1-67 B
1-68 B
1-69 B

Claims

We claim:
1. A compound of formula I:
Figure imgf000114_0001
I
or a pharmaceutically acceptable salt thereof, wherein:
Ring A is a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, phenylene, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
L1 is a covalent bond or a bivalent C1-4 saturated or unsaturated, straight or branched, hydrocarbon chain;
L2 is a covalent bond, or a bivalent C1-4 saturated or unsaturated, straight or branched, hydrocarbon chain, wherein one or two methylene units of the chain are optionally and independently replaced by -C(0) R-, -N(R)C(0)-, - N(R)C(0) R-, - R-, - N(R)S02-, -S02N(R)-, -C(O)-, -OC(O)-, -C(0)0-, -0-, -S-, -SO-, or -S02-;
each R is independently hydrogen, or an optionally substituted group selected from Ci-6 aliphatic, phenyl, 4-7 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or:
two R groups on the same nitrogen are taken together with their intervening atoms to form a 4-7 membered monocyclic saturated, partially unsaturated, or heteroaryl ring having 0-3 heteroatoms, in addition to the nitrogen, independently selected from nitrogen, oxygen, or sulfur;
R1 is hydrogen, optionally substituted Ci-6 aliphatic, or Ring B optionally substituted with 0-4 independently selected Ry groups; Ring B is an optionally substituted group selected from phenyl, 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, a 8-10 membered spirocyclic saturated or partially unsaturated heterocyclic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur,or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each of Rx, Ry, R2 and R3 is independently halogen, R, -OR, -SR, -N(R)2, -C(0)R, - C(0)N(R)2, -N(R)C(0)R, -N(R)C(0)N(R)2, -N(R)C( R)N(R)2, -N02, -CN, - S02N(R)2, -N(R)S02R, -S02N(R)C(0)R, -S02N(R)C(0)N(R)2, S02N(R)C( R)N(R)2 , -C(0)N(R)C(0)R or -CR^O^R; and
m is 0, 1, or 2;
n is 0, 1, 2, 3, or 4;
p is 0 or 1;
X1 is =C(R2)- or =N-;
each of X2 and X3 is -C(R2)= or -N=;
each == is a double bond or a single bond, as valency allows;
Y1 is -N(R3)- or -N=;
Y2 is -C(R3)=, =C(R3)-, =N-, or -N=; and
Y3 is -C= or -N-,
wherein said compound is other than:
Figure imgf000116_0001
Figure imgf000116_0002
2. The compound according to claim 1, wherein Ring A is selected from phenylene, a 4- 7 membered monocyclic saturated or partially unsaturated carbocyclic ring, or a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
3. The compound according to claim 2, wherein Ring A is phenylene, cyclohexylenyl, cyclopentylenyl, or cyclobutylenyl.
4. The compound according to claim 2, wherein Ring A is tetrahydropyranylenyl, piperidinylenyl, or pyrrolidinylenyl.
5. The compound according to any one of claims 1 through 4, wherein L1 is a bivalent Ci-4 saturated or unsaturated, straight or branched, hydrocarbon chain.
6. The compound according to claim 5, wherein L1 is methylene.
7. The compound according to any one of claims 1 through 4, wherein L1 is a covalent bond.
8. The compound according to any one of claims 1 through 7, wherein L2 is -N(R)C(0)-, -N(R)C(0) R-, or -C(0) R-.
9. The compound according to any one of claims 1 through 8, wherein R1 is optionally substituted Ci-6 aliphatic, a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring, a 4-7 membered monocyclic saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 5-6 membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
10. The compound according to claim 9, wherein R1 is oxetanyl, pyrrolidinyl, tetrahydrofuranyl, or tetrahydropyranyl.
11. The compound according to claim 9, wherein R1 is furanyl, oxazolyl, or pyridyl.
12. The compound according to any one of claims 1-8, wherein R1 is optionally substituted Ci-6 aliphatic, phenyl or a 4-7 membered monocyclic saturated or partially unsaturated carbocyclic ring.
13. The compound according to claim 12, wherein R1 is methoxym ethyl.
14. The compound according to claim 12, wherein R1 is cyclopentyl.
15. The compound according to any of claims 1 through 14, wherein R2 is chloro, methoxyl or ethoxyl.
16. The compound according to any of claims 1 through 15, wherein R3 is methyl, - CH2OH, -C(0)N(R)2, or -CH2 H2.
17. The compound according to any of claims 1 through 16, wherein said compound is of formula:
Figure imgf000118_0001
I-A-3
or a pharmaceutically acceptable salt thereof.
18. The compound according to any of claims 1 through 16, wherein said compound is of formula:
Figure imgf000118_0002
I-A-4
a pharmaceutically acceptable salt thereof.
19. The compound according to any of claims 1 throuh 16, wherein said compound is of formula:
Figure imgf000119_0001
The com ound according to claim 1, wherein said compound is of formula:
Figure imgf000119_0002
IV or X
rmaceutically acceptable salt thereof.
Th mpound of claim 1, wherein said compound is of formula:
Figure imgf000119_0003
VI VII
Figure imgf000120_0001
or VIII
or a pharmaceutically acceptable salt thereof.
The compound of claim 1, wherein said compound is of formula:
Figure imgf000120_0002
or IX
or a pharmaceutically acceptable salt thereof.
23. The compound according to claim 1, wherein said compound is selected from those depicted in Table 1.
24. A pharmaceutical composition comprising the compound according to claim 1 and a pharmaceutically acceptable carrier, adjuvant or vehicle.
25. The composition according to claim 20, wherein said composition is administered in combination with an additional therapeutic agent.
26. A method of inhibiting MTHFD2 activity in a biological sample comprising the step of contacting said biological sample with a compound according to any of claims 1 through 22, or a composition thereof.
27. A method of inhibiting MTHFD2 activity in a patient comprising the step of administering to said patient a compound according to any of claims 1 through 22, or a composition thereof.
28. A method for treating a disorder mediated by MTHFD2 in a patient in need thereof, comprising the step of administering to said patient a compound according to any of claims 1 through 22 or pharmaceutically acceptable composition thereof.
29. The method according to claim 28, wherein said disorder mediated by MTHFD2 is a cancer, a carcinoma, or a sarcoma.
30. The method according to claim 29, wherein said cancer is selected from a leukemia, polycythemia vera, a lymphoma, Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain disease, or a solid tumor.
31. The method according to claim 29, wherein the cancer is melanoma, breast cancer, non-small cell lung cancer or hepatocellular carcinoma.
32. The method according to claim 29, wherein said sarcoma or carcinoma is selected from fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, meningioma, melanoma, neuroblastoma, or retinoblastoma.
33. The method according to claim 28, wherein said disorder is a cellular proliferative disorder.
34. The method according to claim 33, wherein said disorder is selected from obesity, benign prostatic hyperplasia, psoriasis, abnormal keratinization, a lymphoproliferative disorder, chronic rheumatoid arthritis, arteriosclerosis, restenosis, and diabetic retinopathy.
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