WO2011151361A1 - Novel compounds - Google Patents

Novel compounds Download PDF

Info

Publication number
WO2011151361A1
WO2011151361A1 PCT/EP2011/059011 EP2011059011W WO2011151361A1 WO 2011151361 A1 WO2011151361 A1 WO 2011151361A1 EP 2011059011 W EP2011059011 W EP 2011059011W WO 2011151361 A1 WO2011151361 A1 WO 2011151361A1
Authority
WO
WIPO (PCT)
Prior art keywords
chloro
compound
mmol
carboxamide
pyridinyl
Prior art date
Application number
PCT/EP2011/059011
Other languages
French (fr)
Inventor
Barbara Bertani
Susanna Cremonesi
Vincenzo Garzya
Fabrizio Micheli
Renata Rupcic
Fabio Maria Sabbatini
Original Assignee
Glaxo Group Limited
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB1009341.7A external-priority patent/GB201009341D0/en
Priority claimed from GBGB1019575.8A external-priority patent/GB201019575D0/en
Priority claimed from GBGB1020622.5A external-priority patent/GB201020622D0/en
Application filed by Glaxo Group Limited filed Critical Glaxo Group Limited
Publication of WO2011151361A1 publication Critical patent/WO2011151361A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system

Definitions

  • the invention relates to tricyclic derivatives, and their use in treating diseases and conditions mediated by antagonism of the mGluRs receptor, in particular substance related disorders.
  • the invention relates to compositions containing the derivatives and processes for their preparation.
  • the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,
  • E is -CH2-, -0-, -S-, -(SO)- or -SO2-;
  • ring A is either:
  • ring A is imidazole
  • RY is H, C-
  • n 1 or 2;
  • R1 is phenyl or 5- or 6-membered monocyclic heteroaryl, either of which are
  • R 2 , R3, R and R 5 which may be the same or different, are selected from the list consisting of H, halo, C-
  • the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,
  • E is -CH2-, -0-, -S-, -(SO)- or -SO2-;
  • ring A is either:
  • ring A is imidazole
  • RY is H, C-
  • n 1 or 2;
  • R1 is phenyl or 5- or 6-membered monocyclic heteroaryl, either of which are
  • halo optionally substituted by one or two groups independently selected from halo, C-
  • R 2 , R3, R4 and R 5 which may be the same or different, are selected from the list consisting of H, halo, C-
  • an alkyl substituent is a univalent radical derived by removal of a hydrogen atom from an acyclic alkane.
  • .galkyl refers to such an alkyl substituent containing 1 to 6 carbons.
  • alkyl substituents include methyl and ethyl, may be straight chain (i.e. n-propyl, n-butyl, n- pentyl and n-hexyl) or branched chain (for example, isopropyl, isobutyl, secbutyl, tert- butyl, isopentyl and neopentyl).
  • such an alkyl substituent is methyl, ethyl, n-propyl or isopropyl.
  • a halo substituent refers to fluoro, chloro, bromo and iodo radicals. In an embodiment unless otherwise indicated such a halo substituent is fluoro or chloro.
  • a haloalkyl substituent is an alkyl group substituted by one or more halo substituents, which halo substituents may be the same or different.
  • .ghaloalkyl refers to such a haloalkyl substituent containing 1 to 6 carbons.
  • Such haloalkyl substituents include monofluoromethyl, difluoromethyl, trifluoromethyl and 1-chloro-2-fluoroethyl.
  • such a haloalkyl substituent is monofluoromethyl, difluoromethyl or trifluoromethyl.
  • an alkoxy substituent is a group of formula " -0-" where R is alkyl as defined above.
  • .galkoxy refers to such an alkoxy substituent containing 1 to 6 carbons.
  • alkoxy substituents include methoxy and ethoxy and may be straight chain (i.e. n-propoxy, n-butoxy, n- pentoxy and n-hexyloxy) or branched chain (for example, isopropoxy, isobutoxy, secbutoxy, tert-butoxy, isopentoxy and neopentoxy).
  • such an alkoxy substituent is methoxy, ethoxy, n-propoxy or isopropoxy.
  • an alkylthio substituent is a group of formula "R-S-" where R is alkyl as defined above.
  • R is alkyl as defined above.
  • .galkylthio refers to such an alkylthio substituent containing 1 to 6 carbons.
  • alkylthio substituents include methylthio and ethylthio and may be straight chain or branched chain.
  • such an alkylthio substituent is methylthio, ethylthio, n-propylthio or isopropylthio.
  • an alkylsulfonyl substituent is a group of formula "R-SO2-" where R is alkyl as defined above.
  • R is alkyl as defined above.
  • .galkylsulfonyl refers to such an alkylsulfonyl substituent containing 1 to 6 carbons.
  • alkylsulfonyl substituents include methylsulfonyl and ethylsulfonyl and may be straight chain or branched chain. In an embodiment unless otherwise indicated, such an alkylsulfonyl substituent is methylsulfonyl, ethylsulfonyl, n-propylsulfonyl or isopropylsulfonyl.
  • a haloalkoxy substituent is a group of formula "R-0-" where R is haloalkyl as defined above.
  • R is haloalkyl as defined above.
  • .ghaloalkoxy refers to such a haloalkoxy substituent containing 1 to 6 carbons.
  • Such haloalkoxy substituents include monofluoromethoxy, difluoromethoxy, trifluoromethoxy and 1 - chloro-2-fluoroethoxy and may be straight chain or branched chain.
  • a haloalkylthio substituent is a group of formula "R-S-" where R is haloalkyl as defined above.
  • .ghaloalkylthio refers to such a haloalkylthio substituent containing 1 to 6 carbons.
  • Such haloalkylthio substituents include monofluoromethylthio, difluoromethylthio, trifluoromethylthio and 1-chloro-2-fluoroethylthio and may be straight chain or branched chain.
  • such a haloalkylthio substituent is monofluoromethylthio, difluoromethylthio or trifluoromethylthio.
  • a haloalkylsulfonyl substituent is a group of formula "R-SO2-" where R is haloalkyl as defined above.
  • .ghaloalkylsulfonyl refers to such a haloalkylsulfonyl substituent containing 1 to 6 carbons.
  • Such haloalkylsulphonyl substituents include monofluoromethylsulfonyl, difluoromethylsulfonyl, trifluoromethylsulfonyl and 1 -chloro-2-fluoroethylsulfonyl and may be straight chain or branched chain.
  • such a haloalkylsulfonyl substituent is monofluoromethylsulfonyl, difluoromethylsulfonyl or trifluoromethylsulfonyl.
  • monocyclic heteroaryl is a univalent radical derived by removal of a hydrogen atom from a monocyclic heteroaromatic ring, which ring comprises ring- carbon atoms and ring-heteroatoms selected from the list nitrogen, oxygen and sulphur, and which ring is aromatic.
  • 5- or 6-membered monocyclic heteroaryl is monocyclic heteroaryl consisting of 5 or 6 ring-atoms, 1 to 3 of which are ring heteroatoms.
  • Examples of monocyclic heteroaryl are furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, azepinyl, oxazepinyl, thiazepinyl and diazepinyl.
  • E is -CH2-, -0-, -S- or -SC>2--
  • ring A is:
  • ring A is imidazole, RY is C-
  • ring A is a triazole ring where V is C, W is C, X is N, Y is N and Z is N.
  • ring A is an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; wherein RY is C-
  • R 1 is pyridyl optionally substituted by halo or alkyl.
  • R5 j hich may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R 2 , R3, R4 and R 5 are H.
  • E is -CH2-, -0-, -S- or -SO2-;
  • ring A is a triazole ring where V is C, W is C, X is N, Y is N and Z is N;
  • n 1 or 2;
  • R1 is pyridyl optionally substituted by halo or alkyl
  • R 2 , R3, R4 and R 5 which may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R 2 , R3 j R4 ANC
  • E is -CH 2 -, -0-, -S- or -SO2-;
  • ring A is an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; wherein RY is C-
  • n 1 or 2;
  • R1 is pyridyl optionally substituted by halo or alkyl
  • R5 J w ich may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R ⁇ , R3, R4 ANC
  • the compound of formula (I) is selected from the list:
  • the pharmaceutically acceptable salts of the compounds defined in the first aspect may contain a basic centre and may form non-toxic acid addition salts formed with inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid, with carboxylic acids or with organo-sulfonic acids.
  • Examples include the HCI, HBr, HI, sulfate or bisulfate, nitrate, phosphate or hydrogen phosphate, acetate, benzoate, succinate, saccharate, fumarate, maleate, lactate, citrate, tartrate, gluconate, camsylate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate salts.
  • suitable pharmaceutical salts see Berge et al, J.
  • pro-drugs for the compounds of the present invention are described in Drugs of Today, Volume 19, Number 9, 1983, pp 499 - 538 and in Topics in Chemistry, Chapter 31 , pp 306 - 316 and in "Design of Prodrugs" by H.
  • the invention provides a prodrug of a compound defined in the first aspect.
  • the compounds defined in the first aspect may exist in solvated or hydrated form. Therefore, in a further aspect, the invention provides a solvate or hydrate of a compound defined in the first aspect or a pharmaceutically acceptable salt thereof.
  • the compounds defined in the first aspect may exist in one or more polymorphic form.
  • the invention provides a polymorph of a compound defined in the first aspect or their pharmaceutically acceptable salts, or a polymorph of a solvate or hydrate of a compound defined in the first aspect or a
  • the compounds of the invention may possess one or more chiral centres and so exist in a number of stereoisomeric forms. All stereoisomers and mixtures thereof are included in the scope of the present invention. Racemic compounds may either be separated using preparative HPLC and a column with a chiral stationary phase or resolved to yield individual enantiomers utilising methods known to those skilled in the art. In addition, chiral intermediate compounds may be resolved and used to prepare chiral compounds of the invention. In addition, the chiral compounds of the invention may be prepared by chiral synthesis. The compounds of the invention may exist in one or more tautomeric forms. All tautomers and mixtures thereof are included in the scope of the present invention. For example, a claim to 2-hydroxyquinolinyl would also cover its tautomeric form, a- quinolinonyl.
  • the invention also includes all suitable isotopic variations of a compound of the invention.
  • An isotopic variation of a compound of the invention is defined as one in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually found in nature.
  • isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine and chlorine such as 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 17 0, 18 0, 35 S, 18 F and 36 CI, respectively.
  • isotopic variations of the invention are useful in drug and/or substrate tissue distribution studies. Tritiated, i.e., 3 H, and carbon-14, i.e., 14 C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with isotopes such as deuterium, i.e., 2 H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements and hence may be preferred in some circumstances.
  • Isotopic variations of the compounds of the invention can generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the Experimental section hereafter using appropriate isotopic variations of suitable reagents.
  • Compound of formula (lib) i.e. compounds of general formula (II) where ring A is an imidazole ring may be prepared according to reaction scheme 3, by reaction compounds of fomula (III) with zinc, followed by reaction with triethylorthoacetate.
  • Compound of general formula (III) can be prepared from compounds of general formula (IV) according to reaction scheme 4.
  • Typical reaction conditions comprise reacting (IV) with Lawesson reagent followed by treatment with base and methyl iodide. The resulting thioether may then be reacted under basic condition with ethyl nitroacetate.
  • Compound of formula (IVb), i.e. compounds of general formula (IVb) where E is - CH 2 -and n is 2, may be prepared according to reaction scheme 6 from compounds of formula (VI). Typical reaction conditions comprise treatment with sodium azide and concentrated sulphuric acid.
  • the compounds of the invention antagonise the mGluR5 receptor and may be used to treat diseases or conditions mediated by antagonism of the mGluR5 receptor. Therefore according to a further aspect, the invention provides a compound of the invention for use in treating a disease or condition.
  • the disease or condition is a human disease or condition.
  • the disease or condition is mediated by antagonism of the mGluR5 receptor.
  • the disease or condition mediated by antagonism of the mGluR5 receptor is selected from the list consisting of: [the numbers in brackets after the listed diseases below refer to the classification code in Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, published by the American Psychiatric Association (DSM-IV) and/or the International Classification of Diseases, 10th Edition (ICD-10)]: i) Psychotic disorders for example Schizophrenia (including the subtypes Paranoid Type (295.30), Disorganised Type (295.10), Catatonic Type (295.20), Undifferentiated Type (295.90) and Residual Type (295.60)); Schizophreniform Disorder (295.40); Schizoaffective Disorder (295.70) (including the subtypes Bipolar Type and Depressive Type); Delusional Disorder (297.1 ) (including the subtypes Erotomanic Type, Grandiose Type, Jealous Type, Persecutory Type, Somatic Type, Mixed Type and Unspecified
  • Psychotic Disorder 297.3
  • Psychotic Disorder due to a General Medical Condition including the subtypes with Delusions and with Hallucinations
  • Substance-Induced Psychotic Disorder including the subtypes with Delusions (293.81 ) and with
  • Depression and mood disorders for example Depressive Episodes (including Major Depressive Episode, Manic Episode, Mixed Episode and Hypomanic Episode); Depressive Disorders (including Major Depressive Disorder, Dysthymic Disorder (300.4), Depressive Disorder Not Otherwise Specified (31 1 )); Bipolar Disorders (including Bipolar I Disorder, Bipolar II Disorder (i.e.
  • Phobia (300.29) (including the subtypes Animal Type, Natural Environment Type, Blood-lnjection-lnjury Type, Situational Type and Other Type); Social Phobia
  • Substance-related disorders for example Substance Use Disorders (including Substance Dependence, Substance Craving and Substance Abuse); Substance- Induced Disorders (including Substance Intoxication, Substance Withdrawal,
  • Alcohol-Induced Persisting Dementia Alcohol-Induced Persisting Amnestic Disorder, Alcohol-Induced Psychotic Disorder, Alcohol-Induced Mood Disorder, Alcohol-Induced Anxiety Disorder, Alcohol-Induced Sexual dysfunction
  • Hallucinogen Dependence (304.50), Hallucinogen Abuse (305.30), Hallucinogen Intoxication (292.89), Hallucinogen Persisting Perception Disorder (Flashbacks)
  • Inhalant-Related Disorders including Inhalant Dependence (304.60), Inhalant Abuse
  • Phencyclidine (or Phencyclidine-Like)-Related Disorders including Phencyclidine Dependence (304.60), Phencyclidine Abuse (305.90), Phencyclidine Intoxication (292.89), Phencyclidine Intoxication Delirium, Phencyclidine-lnduced Psychotic Disorder, Phencyclidine-lnduced Mood Disorder, Phencyclidine-lnduced Anxiety Disorder and Phencyclidine-Related Disorder Not Otherwise Specified (292.9)); Sedative-, Hypnotic-, or Anxiolytic-Related Disorders (including Sedative, Hypnotic, or Anxiolytic Dependence (304.10), Sedative, Hypnotic, or Anxiolytic Abuse (305.40), Sedative, Hypnotic, or Anxiolytic Intoxication (292.89), Sedative, Hypnotic, or Anxiolytic Withdrawal (292.0), Sedative, Hypnotic, or Anxio
  • Eating disorders such as Anorexia Nervosa (307.1 ) (including the subtypes Restricting Type and Binge-Eating/Purging Type); Bulimia Nervosa (307.51 )
  • Attention-Deficit /Hyperactivity Disorder including the subtypes Attention-Deficit /Hyperactivity Disorder Combined Type (314.01 ), Attention-Deficit/Hyperactivity Disorder Predominantly Inattentive Type (314.00), Attention-Deficit/Hyperactivity Disorder Hyperactive-Impulse Type (314.01 ) and Attention-Deficit/Hyperactivity Disorder Not Otherwise Specified (314.9)); Hyperkinetic Disorder; Disruptive
  • Behaviour Disorders such as Conduct Disorder (including the subtypes childhood- onset type (321.81 ), Adolescent-Onset Type (312.82) and Unspecified Onset (312.89), Oppositional Defiant Disorder (313.81 ) and Disruptive Behaviour Disorder Not Otherwise Specified; and Tic Disorders such as Tourette's Disorder (307.23).
  • Enhancement of cognition including the treatment of cognition impairment in other diseases such as schizophrenia, bipolar disorder, depression, other psychiatric disorders and psychotic conditions associated with cognitive impairment, e.g.
  • the disease or condition mediated by antagonism of the mGluR5 receptor is selected from the list consisting of: Parkinson's Disease, epilepsy, inflammatory pain, neuropathic pain, migraine, Down's Syndrome, gastroesophageal reflux disease.
  • the invention provides a compound of the invention for use as a neuroprotectant.
  • the disease or condition mediated by antagonism of the mGluR5 receptor is a substance-related disorder.
  • references herein to "treat”, “treating” or “treatment” extend to prophylaxis, prevention of recurrence and suppression or amelioration of symptoms (whether mild, moderate or severe) as well as the treatment of established conditions.
  • the invention includes the following further aspects. The diseases and conditions described above extend, where appropriate, to these further aspects. i) The use of a compound of the invention in the manufacture of a medicament in treating a disease or condition mediated by antagonism of the mGluR5 receptor.
  • the disease or condition is a substance related disorder.
  • a method of treating a disease or condition mediated by antagonism of the mGluR5 receptor in a mammal (preferably a human) comprising administering an effective amount of a compound of the invention.
  • the disease or condition is a substance related disorder.
  • the compounds of the invention will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient by an appropriate route. Accordingly, in another aspect, the invention provides pharmaceutical compositions comprising a compound of the invention and one or more
  • composition or dosage form other than the compound or compounds of the invention.
  • material usually gives form, consistency and performance to the pharmaceutical
  • compositions of the invention typically contain one compound of the invention. However, in certain embodiments, the pharmaceutical compositions of the invention contain more than one compound of the invention. In addition, the pharmaceutical compositions of the invention may comprise one or more additional pharmaceutically active compounds. Such pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be dispensed and then given to the patient such as with powders or syrups.
  • compositions of the invention may be prepared and packaged as dosage forms wherein each physically discrete dosage form contains a safe and effective amount of a compound of the invention. Accordingly, in another aspect, the invention provides dosage forms comprising pharmaceutical
  • compositions of the invention typically contains from 0.1 mg to 100 mg of a compound of the invention.
  • the compositions of the invention will typically be formulated into dosage forms which are adapted for administration to the patient by the desired route of administration.
  • dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets and cachets; (2) parenteral administration such as sterile solutions, suspensions, implants and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal and vaginal administration such as suppositories, pessaries and foams; (5) inhalation and intranasal such as dry powders, aerosols, suspensions and solutions (sprays and drops); (6) topical administration such as creams, ointments, lotions, solutions, pastes, drops, sprays, foams and gels; (7)
  • Suitable pharmaceutically-acceptable excipients will vary depending upon the particular dosage form chosen.
  • suitable pharmaceutically-acceptable excipients may be chosen for a particular function that they may serve in the composition.
  • certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms.
  • Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms.
  • Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the carrying or transporting of the compound or compounds of the invention once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body.
  • Certain pharmaceutically-acceptable excipients may be chosen for their ability to enhance patient compliance.
  • Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the release of the compound of the invention at the appropriate rate to treat the condition.
  • Suitable pharmaceutically-acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavouring agents, flavour masking agents, colouring agents, anticaking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, rate modifying agents, antioxidants, preservatives, stabilizers, surfactants and buffering agents.
  • excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavouring agents, flavour masking agents, colouring agents, anticaking agents, hume
  • pharmaceutically-acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.
  • Skilled artisans possess the knowledge and skill in the art to enable them to determine suitable pharmaceutically-acceptable excipients in appropriate amounts for use with the compounds of the invention.
  • resources that are available to the skilled artisan which describe pharmaceutically- acceptable excipients and may be useful in selecting suitable pharmaceutically- acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).
  • the pharmaceutical compositions of the invention may be prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).
  • the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of the invention and a diluent or filler.
  • Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate.
  • the oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g.
  • the oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include starches, crospovidone, sodium starch glycolate, cros- carmellose, alginic acid, and sodium carboxymethyl cellulose.
  • the oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and sodium dodecyl sulphate.
  • the oral solid dosage form may further comprise a glidant such as talc and colloidal silicon dioxide.
  • the oral solid dosage form may further comprise an outer coating which may have cosmetic or functional properties.
  • the compounds of the invention may be used in combination with the following agents to treat or prevent psychotic disorders: i) antipsychotics; ii) drugs for extrapyramidal side effects, for example anticholinergics (such as benztropine, biperiden, procyclidine and trihexyphenidyl), antihistamines (such as
  • diphenhydramine and dopaminergics (such as amantadine); iii) antidepressants; iv) anxiolytics; and v) cognitive enhancers for example cholinesterase inhibitors (such as tacrine, donepezil, rivastigmine and galantamine).
  • dopaminergics such as amantadine
  • antidepressants such as amantadine
  • anxiolytics iv
  • cognitive enhancers for example cholinesterase inhibitors (such as tacrine, donepezil, rivastigmine and galantamine).
  • the compounds of the invention may be used in combination with antidepressants to treat or prevent depression and mood disorders.
  • the compounds of the invention may be used in combination with the following agents to treat or prevent bipolar disease: i) mood stabilisers; ii) antipsychotics; and iii) antidepressants.
  • the compounds of the invention may be used in combination with the following agents to treat or prevent anxiety disorders: i) anxiolytics; and ii) antidepressants.
  • Antipsychotic drugs include Typical Antipsychotics (for example chlorpromazine, thioridazine, mesoridazine, fluphenazine, perphenazine, prochlorperazine, trifluoperazine, thiothixine, haloperidol, molindone and loxapine); and Atypical Antipsychotics (for example clozapine, olanzapine, risperidone, quetiapine, aripirazole, ziprasidone and amisulpride).
  • Typical Antipsychotics for example chlorpromazine, thioridazine, mesoridazine, fluphenazine, perphenazine, prochlorperazine, trifluoperazine, thiothixine, haloperidol, molindone and loxapine
  • Atypical Antipsychotics for example clozapine, olanzapine, risperidone, quetiapine,
  • Antidepressant drugs include serotonin reuptake inhibitors (such as citalopram, escitalopram, fluoxetine, paroxetine, sertraline femoxetine, fluvoxamine, indalpine and zimeldine); dual serotonin/noradrenaline reuptake inhibitors (such as venlafaxine, duloxetine and milnacipran); Noradrenaline reuptake inhibitors (such as reboxetine and venlafaxine); tricyclic antidepressants (such as amitriptyline, clomipramine, imipramine, maprotiline, nortriptyline and trimipramine); monoamine oxidase inhibitors (such as isocarboxazide, moclobemide, phenelzine and tranylcypromine); and others (such as bupropion, mianserin, mirtazapine, nefazodone and trazodone).
  • Mood stabiliser drugs include lithium, sodium valproate/valproic acid/divalproex, carbamazepine, lamotrigine, gabapentin, topiramate and tiagabine.
  • Anxiolytics include benzodiazepines such as alprazolam and lorazepam.
  • the compounds of the combination or composition may be administered simultaneously (either in the same or different pharmaceutical formulations), separately or sequentially.
  • the starting material may not necessarily have been prepared from the batch referred to.
  • Mass spectra where recorded using a combined high pressure liquid chromatography/ mass spectroscopy (HPLC/MS) apparatus.
  • HPLC part of the apparatus comprised a Sunfire C18 column (30mm x 4.6 mm i.d.35 urn packing diameter) operated at 30 degC.
  • the compounds were eluted using the following solvent gradient where A is 0.1 % v/v solution of formic acid in water and B is a 0.1 % v/v solution of formic acid in acetonitrile.
  • UV detection comprised a summed signal from wavelength 210 to 350 nm.
  • Toluene (20 mL) was sealed and heated in Biotage Initiator at 150 °C for 7 min. After cooling the solvent was filtered off and the residue was triturated in cyclohexane (ca 100 ml) and methanol (ca 2 ml) and filtered. The title product (1 .1 g) was obtained after drying at 40°C and 0 mmHg for 2 hours in the form of pale green powder. The filtrate was evaporated and triturated once more in the same manner and the title compound (0.35 g) was obtained in the form of pale green powder.
  • reaction mixture was stirred at rt for 22h then triethyl orthoacetate (0.094 mL, 0.508 mmol) was added and reaction was stirred for another 2h. Then it was quenched with water (50ml) and extracted with DCM (5x50ml). The organic layer was washed with NaHCC>3(sat.sol.) until evolution of CO2 ceased. Then it was dried over sodium sulphate/magnesium sulphate and passed through an 1ST Phase separator and evaporated to give a crude oil (387 mg).
  • the organic solvent was removed under vacuum and the resulting aqueous residue was extracted with EtOAc (3 ⁇ 100 mL), and dried over Na 2 S0 4 .
  • the solvent was removed under vacuum to obtain the title compound (3.74 g).
  • the filter cake contained significant amounts of the desired compound and therefore it was transfered to a round bottom flask and triturated with EtOAc (150 mL). The mixture was heated for 1 hour at 60 °C, and filtered hot through a Buchner funnel to receive a colourless solution.
  • Reaction mixture was quenched with water (50 mL) and extracted with DCM (3 x 15 ml_). Combined organic extracts were washed with sat NaHC0 3 (3 x 30 mL), brine (50 mL), and dried over Na 2 S0 4 .
  • Reaction mixture was quenched with water (30 mL) and extracted with DCM (3x30 mL). Organic phase was washed with NaHC03 saturated solution (3x20 mL) and water (1 x30 mL), then dried over Na2S04, filtered and evaporated.
  • the mixture was filtered to give a green precipitate which was dried then washed using warm water (2 litres at 35 °C)
  • the washings were basified with 25 % aqueous ammonia solution to pH 1 1-12 (during base addition the mixture was cooled with ice added to the mixture).
  • 6-Chloro-1 ,3-benzothiazol-2-amine (intermediate 39, 8.7 g, 47.1 mmol) was suspended in 50 % aqueous sodium hydroxide solution (120 mL) and stirred at reflux for 18 hours. Water (50 ml) was added and the mixture was filtered through a Buchner funnel. Adding ice in the mixture and cooling it in the ice bath, the mixture was vigorously stirred and pH of the mixture was adjusted to 6.5 to 7 using glacial acetic acid.
  • the mixture was stirred at room temperature for 2 hours then the reaction mixture was poured into 50 ml of water.
  • the organic substances were extracted using phase separator cartridge using 60 ml of DCM in total.
  • the DCM layer was then washed twice with 100 ml of NaHCC>3 (in total).
  • the precipitate was dissolved in warm water (1 litre) and basified with 25 % aqueous ammonium hydroxide solution to pH 1 1 to 12 with cooling. The resulting precipitate was filtered, washed with aqueous ammonium hydroxide to pH 1 1 to 12) and dried in a vacuum oven at 40 °C to give the title mixture of two regioisomers, 5-chloro-6-fluoro-1 ,3-benzothiazol-2-amine (6.32 g) and 7-chloro-6-fluoro-1 ,3-benzothiazol-2-amine (3.16 g);
  • the mixture was extracted with DCM (200 ml was used in total) and the DCM layer was dried using a phase separator cartridge. The solvent was evaporated and the residue was dissolved in dimethyl sulfoxide (200 mL) and heated at 70 °C for 16 hours. The mixture was cooled to room temperature, poured in 200 ml of water and the resulting precipitate was filtered and dried in a vacuum oven at 40 °C.
  • 6-Chloro-7-fluoro-3-(methylthio)-2H-1 ,4-benzothiazine (intermediate 52, 1.25 g, 5.05 mmol) was dissolved in ethyl nitroacetate (6 ml, 54.2 mmol) under an argon atmosphere.
  • Benzyltrimethylammonium hydroxide in 40 % methanol solution (2.316 ml, 5.10 mmol) was added and the resulting mixture was stirred for 48 hours at 40 °C.
  • the mixture was diluted with water (approximately 100 ml).
  • Organic substances were extracted with 3x50 ml of DCM and the combined DCM layers were washed with 2x50 ml of 10 % aqueous potassium carbonate solution.
  • 6-Fluoro-1 ,3-benzothiazol-2-amine (intermediate 57, 12 g, 71.3 mmol) was suspended in 50 % aquous sodium hydroxide soluiton (200 mL) and heated under reflux for 24 hours. The mixture was cooled, diluted with water (70 mL) and extracted with ethyl acetate (3x). The organic phase was washed with water and all aqueous layers (3 L in total) were combined, filtered and the filtrate was acidified with glacial acetic acid to pH 6). This aqueous mixture was extracted with ethyl-acetate (3x) and the combined extracts were washed with water and dried over IS ⁇ SC ⁇ . The solvent was evaporated to give the title compound (6.2 g); 1 H NMR (500 MHz,
  • the second reaction vessel also containing 7-fluoro-2H-1 ,4-benzothiazin-3(4H)-one (1 ,7 g, 9,28 mmol) and Lawesson's reagent (2,252 g, 5,57 mmol) suspended in toluene (18 mL) was sealed and heated in Biotage Initiator at 150 °C for 7 minutes. After cooling, the two reaction mixtures were gathered and triturated with
  • Triethyl orthoacetate (1 .3 mL, 7.05 mmol) was added followed by p-toluenesulfonic acid monohydrate (14.0 mg, 0.074 mmol) and the mixture was heated at 70 °C with stirring for 2 hours. The reaction was quenched with water and extracted with DCM (2x). Organic phase was washed with saturated aqueous sodium bicarbonate solution, dried using a phase separator filter tube and concentrated under reduced pressure.
  • the reaction was quenched with dioxane/water (30/1 )and the reaction mixture was evaporated to dryness.
  • DCM 50 ml was added and stirred for 0.5 h.
  • the layers were separated by passing through an 1ST Phase separator, and the aqueous layer was extracted three times with DCM (3x50ml). The combined organic layers were then washed with water (100 ml) and brine (100ml), dried over sodium sulphate and passed through an 1ST phase separator.
  • the reaction was then cooled to room temperature and then to 0°C and quenched with dioxane/water (30/1 ). The entire reaction mixture was evaporated to dryness. DCM (50 ml) was added and stirred for 0.5 h. NaHC0 3 (sat. sol.) (20 ml) was added and stirring continued for 15 min. The layers were separated by passing the mixture through an 1ST Phase separator. The aqueous layer was extracted three times with DCM (3x50ml) and the combined organic layers were washed with water (100 ml) and brine (100ml), dried over sodium sulphate and passed through an 1ST phase separator.
  • the combined organic extracts were diluted with water (30 mL) and the pH adjusted to 2 with 0.1 M HCI. The layers were separated and the aqueous layer was extracted with DCM (25 mL). The combined organic extracts were washed with saturated aqueous sodium bicarbonate (30 mL), brine (30 mL) and dried over Na 2 S0 4 .
  • the reaction mixture was cooled to room temperature and carefully quenched with water (3 mL), stirred for 10 minutes and concentrated under vacuum.
  • the residue was diluted with DCM (50 mL) and washed with 0.5 M NaOH (3x30 mL).
  • the combined organic extracts were diluted with water (50 mL) and the pH adjusted to 2.0 with 5 M HCI.
  • the layers were separated and the aqueous layer extracted with DCM (50 mL). Combined organic extracts were washed with brine (50 mL) and dried over Na 2 S0 4 .
  • reaction mixture was diluted with DCM (20 mL) and washed with water (20 mL) and saturated aqueous sodium bicarbonate solution (20 mL). The combined water aqueous layers were extracted with DCM (4x20 mL) and the combined organic extracts were dried over Na2SC> , filtered and evaporated.
  • reaction mixture was cooled to 0 °C and quenched with dioxane/water (30ml_/1 mL) and stirred for 10 min.
  • the reaction mixture was concentrated, dissolved in water (25 mL) and extracted with DCM (4x25 ml). The combined organic extracts were washed with brine (25 ml) and dried over anhydrous Na2SC>4. The solvent was filtered through a phase separator filter tube and removed in vacuo.
  • the mixture was cooled to 0 °C and quenched with dioxane/water (30 mL/1 mL) and stirred for 10 min.
  • the mixture was concentrated and dissolved in water (25 mL) and extracted with DCM (4 x 35 ml).
  • the combined organic extracts were washed with 1 N HCI (3x25 ml), then with brine (25 ml) and dried over anhydrous Na2SC> .
  • the solvent was filtered through a phase separator filter tube and removed in vacuo.
  • reaction mixture was cooled to room temperature, quenched with dioxane/water (2.0/0.5 mL) and stirred for 10 min. 1 M NaOH (15 mL) was added and the mixture was extracted with DCM (4x15 ml). The combined organic extracts were washed with 1 N HCI (4x10 mL) then brine (15 ml), and dried over anhydrous Na2SC> . The mixture was filtered through a phase separator filter tube and the solvent removed in vacuo.
  • the mixture was extracted with DCM (4 x 30 ml) and the combined organic extracts were washed with 1 N HCI (4x25 mL), then brine (20 ml) and dried over anhydrous IS ⁇ SC ⁇ .
  • the mixture was filtered through a phase separator filter tube and the solvent removed in vacuo.
  • the residue was purified using a Biotage SP1 purification system using a Normal phase Silica SNAP 10 g column, eluting with a gradient of EtOAc in Cyclohexane (8 % in 1 CV, 8-66 % in 15 CV, 66 % in 5 CV), followed by precipitation from
  • the retained aqueous phase was basified with 1 M NaOH and extracted with DCM.
  • the organic layer was dried using a phase separator filter tube and concentrated under reduced pressure to give a residue containing the title compound together with some impurities.
  • This residue was dissolved in DCM, 1 M HCI was added and mixture stirred vigorously for 1 hour.
  • the layers were separated and the water layer basified with 1 M NaOH.
  • the mixture was was extracted with DCM and the organic layer dried using a phase separator filter tube and concentrated under reduced pressure to give the title compound of >90% purity (by NMR). This 90% pure material was further purified.
  • the residue was dissolved in DCM, water was added and mixture stirred vigorously for 1 hour.
  • the compounds of the invention may be tested in the following mGluR5 aequorin assay.
  • CHO cells containing human mGluR5 receptors with Tet On expression control technology (supplied by Clontech) were prepared. These cells were grown in cell factories, induced with 10ng/ml doxycycline to enable expression, harvested and then cryo-preserved at -140°C in 1 ml aliquots for future use. On the afternoon prior to the assay, the cells were thawed, suspended in growth media and centrifuged at l OOOrpm for 5 min. The growth media consisted of F12 Hams Nutrient mix (supplied by Gibco - catalogue number 21765) and 10% Tet approved FBS (supplied by Clontech - catalogue number 631 106).
  • the cells were then re-suspended in growth media and incubated at 37°C for 1 hour in a spinner flask. After this post thaw recovery period, the cell suspension was centrifuged once more and resuspended at 2.5 x 10 ⁇ cells/ml in loading buffer consisting of HBSS, 0.1 % BSA (supplied by CalBiochem - catalogue number 126609) and 0.1 % Pluronic F68 (supplied by Gibco - catalogue number 24040-032). The cells were loaded with coelentrazine (supplied by Invitrogen C - catalogue number 6780) to a concentration of 5 ⁇ , wrapped in foil and loaded overnight with mixing. Immediately prior to the assay, the cells were diluted to 15 x 10 ⁇ cells/ml in dilution buffer consisting of HBSS and 0.1 % Pluronic F68.
  • Coelentrazine is the chromophore co-factor which activates the apo-protein, aequorin.
  • the protein has three high affinity binding sites for calcium.
  • agonism of the mGluR5 receptor binding of calcium to the aequorin protein induces a conformational change resulting in an oxidative decarboxylation reaction producing coelenteramide and a flash luminescence signal. This signal was measured using the Lumilux (supplied by Perkin Elmer).
  • test compounds were prepared in DMSO at a concentration of 3mM. These solutions are serially diluted with DMSO to 1 in 4 using a Biomek FX liquid handling device (supplied by Beckman Coulter) in a 384-well compound plate (supplied by Greiner). Daughter plates of 0.5 ⁇ / ⁇ were stamped-out from this master plate for use in the assay. Glutamate Dose Response Curve Preparation
  • a 100mM solution of glutamic acid was prepared in water. This was further diluted with DMSO to a concentration of 10mM. 16 x 1 1 point concentration response curves (CRC) were prepared in DMSO, making the final assay concentration 1.66 x10 " ⁇ M, with 1 in 3 serial dilutions using the Biomek FX. 0.5 ⁇ stamp-outs of this plate were generated for use in the assay.
  • CRC concentration response curves
  • the glutamate CRC plate was placed on the Lumilux where 20 ⁇ / ⁇ of dilution buffer was added, followed by 10 ⁇ /well of loaded cell suspension and a
  • luminescence read was made.
  • An EC80 concentration of glutamate was calculated by using 4X EC50 generated.
  • the EC80 solution was prepared in dilution buffer and added to a reservoir within the Lumilux.

Abstract

The invention relates to tricyclic derivatives, and their use in treating diseases and conditions mediated by antagonism of the mGluR5 receptor, in particular substance related disorders. In addition, the invention relates to compositions containing the derivatives and processes for their preparation.

Description

TITLE OF THE INVENTION
Novel Compounds
BACKGROUND OF THE INVENTION
The invention relates to tricyclic derivatives, and their use in treating diseases and conditions mediated by antagonism of the mGluRs receptor, in particular substance related disorders. In addition, the invention relates to compositions containing the derivatives and processes for their preparation.
BRIEF SUMMARY OF THE INVENTION
According to a first aspect, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,
Figure imgf000002_0001
(I)
wherein
E is -CH2-, -0-, -S-, -(SO)- or -SO2-;
ring A is either:
a) a triazole ring where V is C, W is C, X is N, Y is N and Z is N;
b) an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; or c) an oxazole ring where V is C, W is C, X is C, Y is O and Z is N;
wherein when ring A is imidazole, RY is H, C-|.galkyl, C-|.ghaloalkyl,
C3_gcycloalkyl or C3_gcycloalkyl(C-| _4)alkyl;
n is 1 or 2;
R1 is phenyl or 5- or 6-membered monocyclic heteroaryl, either of which are
optionally substituted by one or two groups independently selected from halo, C-|.galkyl, C-|.galkoxy, C-| .ghaloalkyl, C-|.ghaloalkoxy, cyano, C-| .galkylthio, C-|.ghaloalkylthio, C-| .galkylsulfonyl, C-| .ghaloalkylsulfonyl, C-| .galkylcarbonyl and C-| .ghaloalkylcarbonyl; and R2, R3, R and R5, which may be the same or different, are selected from the list consisting of H, halo, C-|.galkyl, C-|.galkoxy, C-|.ghaloalkyl, C-|.ghaloalkoxy, cyano, C-|.galkylthio and C-|.ghaloalkylthio, wherein two or more of R2, R3, R4 and R5 are H.
DETAILED DESCRIPTION OF THE INVENTION
According to a first aspect, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,
Figure imgf000003_0001
(I)
wherein
E is -CH2-, -0-, -S-, -(SO)- or -SO2-;
ring A is either:
a) a triazole ring where V is C, W is C, X is N, Y is N and Z is N;
b) an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; or c) an oxazole ring where V is C, W is C, X is C, Y is O and Z is N;
wherein when ring A is imidazole, RY is H, C-|.galkyl, C-|.ghaloalkyl,
C3_gcycloalkyl or C3_gcycloalkyl(C-| _4)alkyl;
n is 1 or 2;
R1 is phenyl or 5- or 6-membered monocyclic heteroaryl, either of which are
optionally substituted by one or two groups independently selected from halo, C-|.galkyl, C-|.galkoxy, C-|.ghaloalkyl, C-|.ghaloalkoxy, cyano, C-|.galkylthio, C-|.ghaloalkylthio, C-|.galkylsulfonyl, C-|.ghaloalkylsulfonyl, C-|.galkylcarbonyl and C-|.ghaloalkylcarbonyl; and
R2, R3, R4 and R5, which may be the same or different, are selected from the list consisting of H, halo, C-|.galkyl, C-|.galkoxy, C-|.ghaloalkyl, C-|.ghaloalkoxy, cyano, C-|.galkylthio and C-|.ghaloalkylthio, wherein two or more of R2, R^, R4 and R5 are H. As used herein unless otherwise indicated, an alkyl substituent is a univalent radical derived by removal of a hydrogen atom from an acyclic alkane. For example, C-|.galkyl refers to such an alkyl substituent containing 1 to 6 carbons. Such alkyl substituents include methyl and ethyl, may be straight chain (i.e. n-propyl, n-butyl, n- pentyl and n-hexyl) or branched chain (for example, isopropyl, isobutyl, secbutyl, tert- butyl, isopentyl and neopentyl). In an embodiment unless otherwise indicated, such an alkyl substituent is methyl, ethyl, n-propyl or isopropyl.
As used herein, a halo substituent refers to fluoro, chloro, bromo and iodo radicals. In an embodiment unless otherwise indicated such a halo substituent is fluoro or chloro.
As used herein unless otherwise indicated, a haloalkyl substituent is an alkyl group substituted by one or more halo substituents, which halo substituents may be the same or different. For example, C-|.ghaloalkyl refers to such a haloalkyl substituent containing 1 to 6 carbons. Such haloalkyl substituents include monofluoromethyl, difluoromethyl, trifluoromethyl and 1-chloro-2-fluoroethyl. In an embodiment unless otherwise indicated such a haloalkyl substituent is monofluoromethyl, difluoromethyl or trifluoromethyl. As used herein unless otherwise indicated, an alkoxy substituent is a group of formula " -0-" where R is alkyl as defined above. For example, C-|.galkoxy refers to such an alkoxy substituent containing 1 to 6 carbons. Such alkoxy substituents include methoxy and ethoxy and may be straight chain (i.e. n-propoxy, n-butoxy, n- pentoxy and n-hexyloxy) or branched chain (for example, isopropoxy, isobutoxy, secbutoxy, tert-butoxy, isopentoxy and neopentoxy). In an embodiment unless otherwise indicated, such an alkoxy substituent is methoxy, ethoxy, n-propoxy or isopropoxy.
As used herein unless otherwise indicated, an alkylthio substituent is a group of formula "R-S-" where R is alkyl as defined above. For example, C-|.galkylthio refers to such an alkylthio substituent containing 1 to 6 carbons. Such alkylthio substituents include methylthio and ethylthio and may be straight chain or branched chain. In an embodiment unless otherwise indicated, such an alkylthio substituent is methylthio, ethylthio, n-propylthio or isopropylthio. As used herein unless otherwise indicated, an alkylsulfonyl substituent is a group of formula "R-SO2-" where R is alkyl as defined above. For example, C-| .galkylsulfonyl refers to such an alkylsulfonyl substituent containing 1 to 6 carbons. Such
alkylsulfonyl substituents include methylsulfonyl and ethylsulfonyl and may be straight chain or branched chain. In an embodiment unless otherwise indicated, such an alkylsulfonyl substituent is methylsulfonyl, ethylsulfonyl, n-propylsulfonyl or isopropylsulfonyl.
As used herein unless otherwise indicated, a haloalkoxy substituent is a group of formula "R-0-" where R is haloalkyl as defined above. For example, C-| .ghaloalkoxy refers to such a haloalkoxy substituent containing 1 to 6 carbons. Such haloalkoxy substituents include monofluoromethoxy, difluoromethoxy, trifluoromethoxy and 1 - chloro-2-fluoroethoxy and may be straight chain or branched chain. In an
embodiment unless otherwise indicated, such a haloalkoxy substituent is
monofluoromethoxy, difluoromethoxy or trifluoromethoxy.
As used herein unless otherwise indicated, a haloalkylthio substituent is a group of formula "R-S-" where R is haloalkyl as defined above. For example,
C-|.ghaloalkylthio refers to such a haloalkylthio substituent containing 1 to 6 carbons. Such haloalkylthio substituents include monofluoromethylthio, difluoromethylthio, trifluoromethylthio and 1-chloro-2-fluoroethylthio and may be straight chain or branched chain. In an embodiment unless otherwise indicated, such a haloalkylthio substituent is monofluoromethylthio, difluoromethylthio or trifluoromethylthio. As used herein unless otherwise indicated, a haloalkylsulfonyl substituent is a group of formula "R-SO2-" where R is haloalkyl as defined above. For example,
C-|.ghaloalkylsulfonyl refers to such a haloalkylsulfonyl substituent containing 1 to 6 carbons. Such haloalkylsulphonyl substituents include monofluoromethylsulfonyl, difluoromethylsulfonyl, trifluoromethylsulfonyl and 1 -chloro-2-fluoroethylsulfonyl and may be straight chain or branched chain. In an embodiment unless otherwise indicated, such a haloalkylsulfonyl substituent is monofluoromethylsulfonyl, difluoromethylsulfonyl or trifluoromethylsulfonyl.
As used herein, monocyclic heteroaryl is a univalent radical derived by removal of a hydrogen atom from a monocyclic heteroaromatic ring, which ring comprises ring- carbon atoms and ring-heteroatoms selected from the list nitrogen, oxygen and sulphur, and which ring is aromatic. For example, 5- or 6-membered monocyclic heteroaryl is monocyclic heteroaryl consisting of 5 or 6 ring-atoms, 1 to 3 of which are ring heteroatoms. Examples of monocyclic heteroaryl are furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, azepinyl, oxazepinyl, thiazepinyl and diazepinyl.
In an embodiment, E is -CH2-, -0-, -S- or -SC>2-- In an embodiment, ring A is:
a) a triazole ring where V is C, W is C, X is N, Y is N and Z is N; or
b) an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N;
wherein when ring A is imidazole, RY is C-|.galkyl, C-|.ghaloalkyl or C3_gcycloalkyl. In a further embodiment, ring A is a triazole ring where V is C, W is C, X is N, Y is N and Z is N.
In a still further embodiment, ring A is an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; wherein RY is C-|.galkyl, C-|.ghaloalkyl or C3_gcycloalkyl.
In an embodiment, R1 is pyridyl optionally substituted by halo or alkyl.
In an embodiment, R2, R3j R4 ANC| R5j hich may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R2, R3, R4 and R5 are H.
In an embodiment,
E is -CH2-, -0-, -S- or -SO2-;
ring A is a triazole ring where V is C, W is C, X is N, Y is N and Z is N;
n is 1 or 2;
R1 is pyridyl optionally substituted by halo or alkyl; and
R2, R3, R4 and R5, which may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R2, R3j R4 ANC| R5 are H. In an embodiment,
E is -CH2-, -0-, -S- or -SO2-; ring A is an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; wherein RY is C-|.galkyl, C-|.ghaloalkyl or C3_gcycloalkyl;
n is 1 or 2;
R1 is pyridyl optionally substituted by halo or alkyl; and
R2, R3, R4 ANC| R5J w ich may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R^, R3, R4 ANC| R5 ARE H
In an embodiment, the compound of formula (I) is selected from the list:
8-chloro-N-(2-methyl-4-pyridinyl)-4H-[1 ,2,3]triazolo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 1 );
8-chloro-N-(2-chloro-4-pyridinyl)-4H-[1 ,2,3]triazolo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 2);
8-chloro-1-methyl-/V-(2-methyl-4-pyridinyl)-4/-/-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 3);
8-chloro-1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 4);
8-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 5);
8-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 6);
8-chloro-/V-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4/-/-imidazo[5, 1 - c][1 ,4]benzoxazine-3-carboxamide (Compound 7);
8-chloro-/V-(2-chloro-4-pyridinyl)-7-fluoro-4/-/-[1 ,2,3]triazolo[5, 1-c][1 ,4]benzoxazine-3- carboxamide (Compound 8);
N-(2-chloro-4-pyridinyl)-5,6-dihydro-4H-[1 ,2,3]triazolo[1 ,5-a][1]benzazepine-3- carboxamide (Compound 9);
N-(2-chloro-4-pyridinyl)-1-methyl-5,6-dihydro-4H-imidazo[1 ,5-a][1]benzazepine-3- carboxamide (Compound 10);
N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3-carboxamide (Compound 1 1 );
1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 12);
N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 13);
N-(6-chloro-2-pyridinyl)-1-methyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3-carboxamide 5,5-dioxide (Compound 14); N-(2-chloro-4-pyridinyl)-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxamide (Compound 15);
N-(2-chloro-4-pyridinyl)-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxamide 5,5-dioxide (Compound 16);
N-(2-methyl-4-pyridinyl)-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxamide (Compound 17);
N-(2-chloro-4-pyridinyl)-1-(trifluoromethyl)-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3- carboxamide (Compound 18);
N-(2-chloro-4-pyridinyl)-1-cyclopropyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 19);
N-(2-chloro-4-pyridinyl)-7-f luoro-4H-[1 ,2,3]triazolo[5, 1 -c][1 ,4]benzoxazine-3- carboxamide (Compound 20);
N-(6-chloro-2-pyridinyl)-7-f luoro-4H-[1 ,2,3]triazolo[5, 1 -c][1 ,4]benzoxazine-3- carboxamide (Compound 21 );
N-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazo[5,1 -c][1 ,4]benzoxazine-3- carboxamide (Compound 22);
7-chloro-N-(2-chloro-4-pyridinyl)-4H-[1 !2,3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide (Compound 23);
7-chloro-N-(2-methyl-4-pyridinyl)-4H-[1 !2!3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide (Compound 24);
7-chloro-N-(2-chloro-4-pyridinyl)-1 -methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3- carboxamide (Compound 25);
7- chloro-N-(2-chloro-4-pyridinyl)-4H-[1 !2,3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 26);
7-chloro-1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5,1-c][1 ,4]benzothiazine-3- carboxamide (Compound 27);
8- chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine- 3-carboxamide (Compound 28);
8-chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazo[5,1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 29);
8-chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazo[5,1 - c][1 ,4]benzothiazine-3-carboxamide 5,5-dioxide (Compound 30);
N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3- carboxamide (Compound 31 );
7-fluoro-N-(2-methyl-4-pyridinyl)-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3- carboxamide (Compound 32); N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 !2!3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 33);
N-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3- carboxamide (Compound 34);
N-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 35);
7-fluoro-1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5,1-c][1 ,4]benzothiazine-3- carboxamide (Compound 36);
6-chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine- 3-carboxamide (Compound 37); and
6-chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazo[5,1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 38);
or a pharmaceutically acceptable salt thereof. The pharmaceutically acceptable salts of the compounds defined in the first aspect may contain a basic centre and may form non-toxic acid addition salts formed with inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid, with carboxylic acids or with organo-sulfonic acids. Examples include the HCI, HBr, HI, sulfate or bisulfate, nitrate, phosphate or hydrogen phosphate, acetate, benzoate, succinate, saccharate, fumarate, maleate, lactate, citrate, tartrate, gluconate, camsylate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate salts. For reviews on suitable pharmaceutical salts see Berge et al, J. Pharm, Sci., 66, 1-19, 1977; P L Gould, International Journal of Pharmaceutics, 33 (1986), 201 -217; and Bighley et al, Encyclopedia of Pharmaceutical Technology, Marcel Dekker Inc, New York 1996, Volume 13, page 453-497.
It will be appreciated by those skilled in the art that certain protected derivatives of the compounds defined in the first aspect, which may be made prior to a final deprotection stage, may not possess pharmacological activity as such, but may, in certain instances, be administered orally or parenterally and thereafter metabolised in the body to form compounds defined in the first aspect which are pharmacologically active. Such derivatives may therefore be described as "prodrugs". All protected derivatives and prodrugs of compounds defined in the first aspect are included within the scope of the invention. Examples of suitable pro-drugs for the compounds of the present invention are described in Drugs of Today, Volume 19, Number 9, 1983, pp 499 - 538 and in Topics in Chemistry, Chapter 31 , pp 306 - 316 and in "Design of Prodrugs" by H.
Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as "pro-moieties", for example as described by H. Bundgaard in "Design of Prodrugs" (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within the compounds defined in the first aspect. Therefore, in a further aspect, the invention provides a prodrug of a compound defined in the first aspect.
The compounds defined in the first aspect, their pharmaceutically acceptable salts or prodrugs, may exist in solvated or hydrated form. Therefore, in a further aspect, the invention provides a solvate or hydrate of a compound defined in the first aspect or a pharmaceutically acceptable salt thereof.
The compounds defined in the first aspect, their pharmaceutically acceptable salts, or solvates or hydrates of either, may exist in one or more polymorphic form.
Therefore, in a further aspect, the invention provides a polymorph of a compound defined in the first aspect or their pharmaceutically acceptable salts, or a polymorph of a solvate or hydrate of a compound defined in the first aspect or a
pharmaceutically acceptable salt thereof.
Hereinafter, compounds defined in the first aspect, their salts and prodrugs; any solvates or hydrates of any salt or prodrug; and any polymorph of any compound, salt, solvate or hydrate are referred to as "compounds of the invention". The term "compounds of the invention" also includes all embodiments described for the first aspect.
The compounds of the invention may possess one or more chiral centres and so exist in a number of stereoisomeric forms. All stereoisomers and mixtures thereof are included in the scope of the present invention. Racemic compounds may either be separated using preparative HPLC and a column with a chiral stationary phase or resolved to yield individual enantiomers utilising methods known to those skilled in the art. In addition, chiral intermediate compounds may be resolved and used to prepare chiral compounds of the invention. In addition, the chiral compounds of the invention may be prepared by chiral synthesis. The compounds of the invention may exist in one or more tautomeric forms. All tautomers and mixtures thereof are included in the scope of the present invention. For example, a claim to 2-hydroxyquinolinyl would also cover its tautomeric form, a- quinolinonyl.
The invention also includes all suitable isotopic variations of a compound of the invention. An isotopic variation of a compound of the invention is defined as one in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine and chlorine such as 2H, 3H, 11C, 13C, 14C, 15N, 170, 180, 35S, 18F and 36CI, respectively. Certain isotopic variations of the invention, for example, those in which a radioactive isotope such as 3H or 14C is incorporated, are useful in drug and/or substrate tissue distribution studies. Tritiated, i.e., 3H, and carbon-14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with isotopes such as deuterium, i.e., 2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements and hence may be preferred in some circumstances. Isotopic variations of the compounds of the invention can generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the Experimental section hereafter using appropriate isotopic variations of suitable reagents.
Compounds of the invention may be prepared in a variety of ways. In the following reaction schemes and hereafter, unless otherwise stated to R^, ring A, E and n are as defined in the first aspect. These processes form further aspects of the invention.
Throughout the specification, general formulae are designated by Roman numerals (I), (II), (III), (IV) etc. Subsets of these general formulae are defined as (la), (lb), (lc) etc .... (IVa), (IVb), (IVc) etc. Compound of general formula I can be prepared from compounds of general formula II (where R is lower alkyl such as methyl, ethyl, etc) according to reaction scheme 1 by reacting compounds of formula I with an appropriate heteroaromatic amine in the presence of triethyl aluminium.
Scheme 1
Figure imgf000012_0001
(ll) (I)
Compound of formula (I la), i.e. compounds of general formula (II) where ring A is a triazole may be prepared according to reaction scheme 2, by reaction compounds of fomula (III) with zinc, followed by reaction with isoamyl nitrite and trichloroacetic acid. Scheme 2
Figure imgf000012_0002
(lla)
Compound of formula (lib), i.e. compounds of general formula (II) where ring A is an imidazole ring may be prepared according to reaction scheme 3, by reaction compounds of fomula (III) with zinc, followed by reaction with triethylorthoacetate.
Scheme 3
Figure imgf000012_0003
Compound of general formula (III) can be prepared from compounds of general formula (IV) according to reaction scheme 4. Typical reaction conditions comprise reacting (IV) with Lawesson reagent followed by treatment with base and methyl iodide. The resulting thioether may then be reacted under basic condition with ethyl nitroacetate.
Scheme 4
Figure imgf000013_0001
Compounds of formula (IVa), i.e. compounds of general formula (IV) where E is O S and n is 1 , may be prepared according to reaction scheme 5, by reacting compounds of formula (V) with ethylbromoacetate under basic conditions. The resulting compounds may then be cyclised to compounds of formula (IVa) using ammonium chloride and iron.
Scheme 5
Figure imgf000013_0002
Compound of formula (IVb), i.e. compounds of general formula (IVb) where E is - CH2-and n is 2, may be prepared according to reaction scheme 6 from compounds of formula (VI). Typical reaction conditions comprise treatment with sodium azide and concentrated sulphuric acid.
Scheme 6
Figure imgf000014_0001
(VI) (IVb)
Compounds of formula (la), i.e. compounds of formula (I) where E is -S02- may be prepared from compounds of formula (lb) where E is -S-, by treatment with metachloroperbenzoic acid (MCPBA) according to reaction scheme 7.
Scheme 7
Figure imgf000014_0002
It will be appreciated by the skilled chemist that compounds of formula (I) may be converted to other compounds of formula (I) by methods known in the art.
The compounds of the invention antagonise the mGluR5 receptor and may be used to treat diseases or conditions mediated by antagonism of the mGluR5 receptor. Therefore according to a further aspect, the invention provides a compound of the invention for use in treating a disease or condition. In an embodiment the disease or condition is a human disease or condition. In an embodiment the disease or condition is mediated by antagonism of the mGluR5 receptor. In an embodiment the disease or condition mediated by antagonism of the mGluR5 receptor is selected from the list consisting of: [the numbers in brackets after the listed diseases below refer to the classification code in Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, published by the American Psychiatric Association (DSM-IV) and/or the International Classification of Diseases, 10th Edition (ICD-10)]: i) Psychotic disorders for example Schizophrenia (including the subtypes Paranoid Type (295.30), Disorganised Type (295.10), Catatonic Type (295.20), Undifferentiated Type (295.90) and Residual Type (295.60)); Schizophreniform Disorder (295.40); Schizoaffective Disorder (295.70) (including the subtypes Bipolar Type and Depressive Type); Delusional Disorder (297.1 ) (including the subtypes Erotomanic Type, Grandiose Type, Jealous Type, Persecutory Type, Somatic Type, Mixed Type and Unspecified Type); Brief Psychotic Disorder (298.8); Shared
Psychotic Disorder (297.3); Psychotic Disorder due to a General Medical Condition (including the subtypes with Delusions and with Hallucinations); Substance-Induced Psychotic Disorder (including the subtypes with Delusions (293.81 ) and with
Hallucinations (293.82)); and Psychotic Disorder Not Otherwise Specified (298.9). ii) Depression and mood disorders for example Depressive Episodes (including Major Depressive Episode, Manic Episode, Mixed Episode and Hypomanic Episode); Depressive Disorders (including Major Depressive Disorder, Dysthymic Disorder (300.4), Depressive Disorder Not Otherwise Specified (31 1 )); Bipolar Disorders (including Bipolar I Disorder, Bipolar II Disorder (i.e. Recurrent Major Depressive Episodes with Hypomanic Episodes) (296.89), Cyclothymic Disorder (301.13) and Bipolar Disorder Not Otherwise Specified (296.80)); Other Mood Disorders (including Mood Disorder due to a General Medical Condition (293.83) which includes the subtypes With Depressive Features, With Major Depressive-like Episode, With Manic Features and With Mixed Features); Substance-Induced Mood Disorder (including the subtypes With Depressive Features, With Manic Features and With Mixed Features); and Mood Disorder Not Otherwise Specified (296.90). iii) Anxiety disorders for example Social Anxiety Disorder; Panic Attack; Agoraphobia, Panic Disorder; Agoraphobia Without History of Panic Disorder (300.22); Specific
Phobia (300.29) (including the subtypes Animal Type, Natural Environment Type, Blood-lnjection-lnjury Type, Situational Type and Other Type); Social Phobia
(300.23); Obsessive-Compulsive Disorder (300.3); Posttraumatic Stress Disorder (309.81 ); Acute Stress Disorder (308.3); Generalized Anxiety Disorder (300.02); Anxiety Disorder Due to a General Medical Condition (293.84); Substance-Induced Anxiety Disorder; and Anxiety Disorder Not Otherwise Specified (300.00). iv) Substance-related disorders for example Substance Use Disorders (including Substance Dependence, Substance Craving and Substance Abuse); Substance- Induced Disorders (including Substance Intoxication, Substance Withdrawal,
Substance-Induced Delirium, Substance-Induced Persisting Dementia, Substance- Induced Persisting Amnestic Disorder, Substance-Induced Psychotic Disorder, Substance-Induced Mood Disorder, Substance-Induced Anxiety Disorder,
Substance-Induced Sexual Dysfunction, Substance-Induced Sleep Disorder and Hallucinogen Persisting Perception Disorder (Flashbacks); Alcohol-Related
Disorders (including Alcohol Dependence (303.90), Alcohol Abuse (305.00), Alcohol Intoxication (303.00), Alcohol Withdrawal (291.81 ), Alcohol Intoxication Delirium, Alcohol Withdrawal Delirium, Alcohol-Induced Persisting Dementia, Alcohol-Induced Persisting Amnestic Disorder, Alcohol-Induced Psychotic Disorder, Alcohol-Induced Mood Disorder, Alcohol-Induced Anxiety Disorder, Alcohol-Induced Sexual
Dysfunction, Alcohol-Induced Sleep Disorder and Alcohol-Related Disorder Not Otherwise Specified (291 .9)); Amphetamine (or Amphetamine-Like)-Related
Disorders (for example Amphetamine Dependence (304.40), Amphetamine Abuse (305.70), Amphetamine Intoxication (292.89), Amphetamine Withdrawal (292.0), Amphetamine Intoxication Delirium, Amphetamine Induced Psychotic Disorder, Amphetamine-Induced Mood Disorder, Amphetamine-Induced Anxiety Disorder, Amphetamine-Induced Sexual Dysfunction, Amphetamine-Induced Sleep Disorder and Amphetamine-Related Disorder Not Otherwise Specified (292.9)); Caffeine Related Disorders (including Caffeine Intoxication (305.90), Caffeine-Induced Anxiety Disorder, Caffeine-Induced Sleep Disorder and Caffeine-Related Disorder Not Otherwise Specified (292.9)); Cannabis-Related Disorders (including Cannabis Dependence (304.30), Cannabis Abuse (305.20), Cannabis Intoxication (292.89), Cannabis Intoxication Delirium, Cannabis-lnduced Psychotic Disorder, Cannabis- Induced Anxiety Disorder and Cannabis-Related Disorder Not Otherwise Specified (292.9)); Cocaine-Related Disorders (including Cocaine Dependence (304.20), Cocaine Abuse (305.60), Cocaine Intoxication (292.89), Cocaine Withdrawal (292.0), Cocaine Intoxication Delirium, Cocaine-Induced Psychotic Disorder, Cocaine- Induced Mood Disorder, Cocaine-Induced Anxiety Disorder, Cocaine-Induced Sexual Dysfunction, Cocaine-Induced Sleep Disorder and Cocaine-Related Disorder Not Otherwise Specified (292.9)); Hallucinogen-Related Disorders (including
Hallucinogen Dependence (304.50), Hallucinogen Abuse (305.30), Hallucinogen Intoxication (292.89), Hallucinogen Persisting Perception Disorder (Flashbacks)
(292.89) , Hallucinogen Intoxication Delirium, Hallucinogen-Induced Psychotic Disorder, Hallucinogen-Induced Mood Disorder, Hallucinogen-Induced Anxiety Disorder and Hallucinogen-Related Disorder Not Otherwise Specified (292.9));
Inhalant-Related Disorders (including Inhalant Dependence (304.60), Inhalant Abuse
(305.90) , Inhalant Intoxication (292.89), Inhalant Intoxication Delirium, Inhalant- Induced Persisting Dementia, Inhalant-Induced Psychotic Disorder, Inhalant-Induced Mood Disorder, Inhalant-Induced Anxiety Disorder and Inhalant-Related Disorder Not Otherwise Specified (292.9)); Nicotine-Related Disorders (including Nicotine
Dependence (305.1 ), Nicotine Withdrawal (292.0) and Nicotine-Related Disorder Not Otherwise Specified (292.9)); Opioid-Related Disorders (including Opioid
Dependence (304.00), Opioid Abuse (305.50), Opioid Intoxication (292.89), Opioid Withdrawal (292.0), Opioid Intoxication Delirium, Opioid-lnduced Psychotic Disorder, Opioid-lnduced Mood Disorder, Opioid-lnduced Sexual Dysfunction, Opioid-lnduced Sleep Disorder and Opioid-Related Disorder Not Otherwise Specified (292.9));
Phencyclidine (or Phencyclidine-Like)-Related Disorders (including Phencyclidine Dependence (304.60), Phencyclidine Abuse (305.90), Phencyclidine Intoxication (292.89), Phencyclidine Intoxication Delirium, Phencyclidine-lnduced Psychotic Disorder, Phencyclidine-lnduced Mood Disorder, Phencyclidine-lnduced Anxiety Disorder and Phencyclidine-Related Disorder Not Otherwise Specified (292.9)); Sedative-, Hypnotic-, or Anxiolytic-Related Disorders (including Sedative, Hypnotic, or Anxiolytic Dependence (304.10), Sedative, Hypnotic, or Anxiolytic Abuse (305.40), Sedative, Hypnotic, or Anxiolytic Intoxication (292.89), Sedative, Hypnotic, or Anxiolytic Withdrawal (292.0), Sedative, Hypnotic, or Anxiolytic Intoxication Delirium, Sedative, Hypnotic, or Anxiolytic Withdrawal Delirium, Sedative-, Hypnotic-, or Anxiolytic-Persisting Dementia, Sedative-, Hypnotic-, or Anxiolytic- Persisting
Amnestic Disorder, Sedative-, Hypnotic-, or Anxiolytic-lnduced Psychotic Disorder, Sedative-, Hypnotic-, or Anxiolytic-lnduced Mood Disorder, Sedative-, Hypnotic-, or Anxiolytic-lnduced Anxiety Disorder Sedative-, Hypnotic-, or Anxiolytic-lnduced Sexual Dysfunction, Sedative-, Hypnotic-, or Anxiolytic-lnduced Sleep Disorder and Sedative-, Hypnotic-, or Anxiolytic-Related Disorder Not Otherwise Specified
(292.9)); Polysubstance-Related Disorder (including Polysubstance Dependence (304.80)); and Other (or Unknown) Substance-Related Disorders (including Anabolic Steroids, Nitrate Inhalants and Nitrous Oxide). v) Sleep disorders for example primary sleep disorders such as Dyssomnias
(including Primary Insomnia (307.42), Primary Hypersomnia (307.44), Narcolepsy (347), Breathing-Related Sleep Disorders (780.59), Circadian Rhythm Sleep
Disorder (307.45) and Dyssomnia Not Otherwise Specified (307.47)); primary sleep disorders such as Parasomnias (including Nightmare Disorder (307.47), Sleep Terror Disorder (307.46), Sleepwalking Disorder (307.46) and Parasomnia Not Otherwise Specified (307.47)); Sleep Disorders Related to Another Mental Disorder (including Insomnia Related to Another Mental Disorder (307.42) and Hypersomnia Related to Another Mental Disorder (307.44)); Sleep Disorder Due to a General Medical Condition; and Substance-Induced Sleep Disorder (including the subtypes Insomnia Type, Hypersomnia Type, Parasomnia Type and Mixed Type). vi) Eating disorders such as Anorexia Nervosa (307.1 ) (including the subtypes Restricting Type and Binge-Eating/Purging Type); Bulimia Nervosa (307.51 )
(including the subtypes Purging Type and Nonpurging Type); Obesity; Compulsive Eating Disorder; Binge Eating Disorder; and Eating Disorder Not Otherwise Specified (307.50). vii) Autism Spectrum Disorders including Autistic Disorder (299.00), Asperger's Disorder, Rett's Disorder, Childhood Disintegrative Disorder and Pervasive
Developmental Disorder Not Otherwise Specified. viii) Attention-Deficit /Hyperactivity Disorder (including the subtypes Attention-Deficit /Hyperactivity Disorder Combined Type (314.01 ), Attention-Deficit/Hyperactivity Disorder Predominantly Inattentive Type (314.00), Attention-Deficit/Hyperactivity Disorder Hyperactive-Impulse Type (314.01 ) and Attention-Deficit/Hyperactivity Disorder Not Otherwise Specified (314.9)); Hyperkinetic Disorder; Disruptive
Behaviour Disorders such as Conduct Disorder (including the subtypes childhood- onset type (321.81 ), Adolescent-Onset Type (312.82) and Unspecified Onset (312.89), Oppositional Defiant Disorder (313.81 ) and Disruptive Behaviour Disorder Not Otherwise Specified; and Tic Disorders such as Tourette's Disorder (307.23). ix) Enhancement of cognition including the treatment of cognition impairment in other diseases such as schizophrenia, bipolar disorder, depression, other psychiatric disorders and psychotic conditions associated with cognitive impairment, e.g.
Alzheimer's disease.
In an embodiment the disease or condition mediated by antagonism of the mGluR5 receptor is selected from the list consisting of: Parkinson's Disease, epilepsy, inflammatory pain, neuropathic pain, migraine, Down's Syndrome, gastroesophageal reflux disease.
In a further embodiment, the invention provides a compound of the invention for use as a neuroprotectant. In an embodiment the disease or condition mediated by antagonism of the mGluR5 receptor is a substance-related disorder. It will be appreciated that references herein to "treat", "treating" or "treatment" extend to prophylaxis, prevention of recurrence and suppression or amelioration of symptoms (whether mild, moderate or severe) as well as the treatment of established conditions. It will be appreciated that the invention includes the following further aspects. The diseases and conditions described above extend, where appropriate, to these further aspects. i) The use of a compound of the invention in the manufacture of a medicament in treating a disease or condition mediated by antagonism of the mGluR5 receptor. In an embodiment the disease or condition is a substance related disorder. ii) A method of treating a disease or condition mediated by antagonism of the mGluR5 receptor in a mammal (preferably a human) comprising administering an effective amount of a compound of the invention. In an embodiment the disease or condition is a substance related disorder.
The compounds of the invention will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient by an appropriate route. Accordingly, in another aspect, the invention provides pharmaceutical compositions comprising a compound of the invention and one or more
pharmaceutically-acceptable excipients.
As used herein, "pharmaceutically-acceptable excipient" means any
pharmaceutically acceptable material present in the pharmaceutical composition or dosage form other than the compound or compounds of the invention. Typically the material gives form, consistency and performance to the pharmaceutical
composition.
The pharmaceutical compositions of the invention typically contain one compound of the invention. However, in certain embodiments, the pharmaceutical compositions of the invention contain more than one compound of the invention. In addition, the pharmaceutical compositions of the invention may comprise one or more additional pharmaceutically active compounds. Such pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be dispensed and then given to the patient such as with powders or syrups.
Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged as dosage forms wherein each physically discrete dosage form contains a safe and effective amount of a compound of the invention. Accordingly, in another aspect, the invention provides dosage forms comprising pharmaceutical
compositions of the invention. Each discrete dosage form typically contains from 0.1 mg to 100 mg of a compound of the invention. The compositions of the invention will typically be formulated into dosage forms which are adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets and cachets; (2) parenteral administration such as sterile solutions, suspensions, implants and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal and vaginal administration such as suppositories, pessaries and foams; (5) inhalation and intranasal such as dry powders, aerosols, suspensions and solutions (sprays and drops); (6) topical administration such as creams, ointments, lotions, solutions, pastes, drops, sprays, foams and gels; (7) ocular administration such as drops, ointment, sprays, suspensions and inserts; (8) buccal and sublingual administration such as lozenges, patches, sprays, drops, chewing gums and tablets.
Suitable pharmaceutically-acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically-acceptable excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the carrying or transporting of the compound or compounds of the invention once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body. Certain pharmaceutically-acceptable excipients may be chosen for their ability to enhance patient compliance. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the release of the compound of the invention at the appropriate rate to treat the condition.
Suitable pharmaceutically-acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavouring agents, flavour masking agents, colouring agents, anticaking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, rate modifying agents, antioxidants, preservatives, stabilizers, surfactants and buffering agents. The skilled artisan will appreciate that certain
pharmaceutically-acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.
Skilled artisans possess the knowledge and skill in the art to enable them to determine suitable pharmaceutically-acceptable excipients in appropriate amounts for use with the compounds of the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically- acceptable excipients and may be useful in selecting suitable pharmaceutically- acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press). The pharmaceutical compositions of the invention may be prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).
In one aspect, the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of the invention and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g. corn starch, potato starch and pre- gelatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g. hydroxypropyl methyl cellulose). The oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include starches, crospovidone, sodium starch glycolate, cros- carmellose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and sodium dodecyl sulphate. The oral solid dosage form may further comprise a glidant such as talc and colloidal silicon dioxide. The oral solid dosage form may further comprise an outer coating which may have cosmetic or functional properties.
The compounds of the invention may be used in combination with the following agents to treat or prevent psychotic disorders: i) antipsychotics; ii) drugs for extrapyramidal side effects, for example anticholinergics (such as benztropine, biperiden, procyclidine and trihexyphenidyl), antihistamines (such as
diphenhydramine) and dopaminergics (such as amantadine); iii) antidepressants; iv) anxiolytics; and v) cognitive enhancers for example cholinesterase inhibitors (such as tacrine, donepezil, rivastigmine and galantamine).
The compounds of the invention may be used in combination with antidepressants to treat or prevent depression and mood disorders. The compounds of the invention may be used in combination with the following agents to treat or prevent bipolar disease: i) mood stabilisers; ii) antipsychotics; and iii) antidepressants.
The compounds of the invention may be used in combination with the following agents to treat or prevent anxiety disorders: i) anxiolytics; and ii) antidepressants.
Antipsychotic drugs include Typical Antipsychotics (for example chlorpromazine, thioridazine, mesoridazine, fluphenazine, perphenazine, prochlorperazine, trifluoperazine, thiothixine, haloperidol, molindone and loxapine); and Atypical Antipsychotics (for example clozapine, olanzapine, risperidone, quetiapine, aripirazole, ziprasidone and amisulpride). Antidepressant drugs include serotonin reuptake inhibitors (such as citalopram, escitalopram, fluoxetine, paroxetine, sertraline femoxetine, fluvoxamine, indalpine and zimeldine); dual serotonin/noradrenaline reuptake inhibitors (such as venlafaxine, duloxetine and milnacipran); Noradrenaline reuptake inhibitors (such as reboxetine and venlafaxine); tricyclic antidepressants (such as amitriptyline, clomipramine, imipramine, maprotiline, nortriptyline and trimipramine); monoamine oxidase inhibitors (such as isocarboxazide, moclobemide, phenelzine and tranylcypromine); and others (such as bupropion, mianserin, mirtazapine, nefazodone and trazodone).
Mood stabiliser drugs include lithium, sodium valproate/valproic acid/divalproex, carbamazepine, lamotrigine, gabapentin, topiramate and tiagabine. Anxiolytics include benzodiazepines such as alprazolam and lorazepam.
It will be appreciated that the compounds of the combination or composition may be administered simultaneously (either in the same or different pharmaceutical formulations), separately or sequentially.
Supporting Compounds
The preparation of a number of the compounds of the invention are described below.
In the procedures that follow, after each starting material, reference to an
intermediate is typically provided. This is provided merely for assistance to the skilled chemist. The starting material may not necessarily have been prepared from the batch referred to.
Where reference is made to the use of a "similar" or "analogous" procedure, as will be appreciated by those skilled in the art, such a procedure may involve minor variation, for example reaction temperature, reagent/solvent amount, reaction time, work-up conditions or chromatographic purification conditions.
Compounds of the invention and intermediates have been named using ACD/Name PRO 6.02 chemical naming software (Advanced Chemistry Development Inc., Toronto, Ontario, M5H2L3, Canada). Abbreviations
DIPEA - Diisopropylethylamine
mCPBA - metaChloroPerbenzoic Acid
MDAP - Mass Spec. Directed Preparative HPLC purification
HATU - o-(7-azabenzotriazol-1 -yl)-N,N,N',N'- tetramethyluronium
hexafluorophosphate
HOAt - 1-hydroxy-7-azabenzotriazole
EtOAc - Ethylacetate
NMP - N-methyl-2-pyrrolidone
NMR - Nuclear Magnetic Resonance
Physical Characterisation
1 H NMR spectra were run on a Bruker Avance 400MHz instrument with a 5mm QNP probe or a Bruker DPX-250 250MHz instrument with a 5mm QNP probe at 27 C. Data were recorded in deuterio-chloroform, D6-dimethylsulphoxide or in D4-deuterio- methanol with tetramethylsilane (TMS) as an internal reference. Chemical shifts given in parts per million (ppm) relative to TMS. Multiplicities are designated as s (singlet), d (doublet), t (triplet), dd (double-doublet), ddd (double-double-doublet), dt (double triplet), m (multiplet).
Mass spectra where recorded using a combined high pressure liquid chromatography/ mass spectroscopy (HPLC/MS) apparatus. The HPLC part of the apparatus comprised a Sunfire C18 column (30mm x 4.6 mm i.d.35 urn packing diameter) operated at 30 degC. The compounds were eluted using the following solvent gradient where A is 0.1 % v/v solution of formic acid in water and B is a 0.1 % v/v solution of formic acid in acetonitrile. UV detection comprised a summed signal from wavelength 210 to 350 nm.
Time Flow Rate %A %B
(min) (ml/min)
0 3 97 3
0.1 3 97 3
4.2 3 0 100
4.8 3 0 100 4.9 3 97 3
5.0 3 97 3
Intermediate 1 : 6-chloro-2/-/-1 ,4-benzothiazin-3(4/-/)-one
Figure imgf000025_0001
To a solution of 2-amino-4-chlorobenzenethiol (5 g, 31.3 mmol) in dimethyl Sulfoxide (DMSO) (100 mL) stirred in air at rt was added solid sodium ethoxide (2.345 g, 34.5 mmol) and the reaction mixture was stirred at rt for 1 hr. After that methyl bromoacetate (3.17 mL, 34.5 mmol) was added in one portion and stirring was continued for 12h. Then the reaction mixture was divided into 5 portions and each portion added to a 10-20 ml vessel. Each vessel was sealed and heated in a Biotage Initiator at 150 °C for 5 min. After cooling the reaction mixture was evaluated by
UPLC-MS/UV o show only one product in UV (rt - 4.16 min; M+ - 200). The reaction mixture was diluted with ice-cold water (300 ml) and acidified to pH - 5 with 3N HCI. A tan precipitate formed which was filtered and dried at 40 °C under 0 mmHg for 3h; 1H NMR (300 MHz, DMSO-d6 δ ppm 10.7 (s, 1 H), 7.27 - 7.44 (m,1 H), 6.95 - 7.04 (m,2H), 3.44 - 3.5 (s,2H).
Intermediate 2: 6-chloro-2/-/-1 ,4-benzothiazine-3(4/-/)-thione
Figure imgf000025_0002
The reaction vessel containing 6-chloro-2H-1 ,4-benzothiazin-3(4H)-one (intermediate 1 ) (2 g, 10.02 mmol) and Lawesson's reagent (2.84 g, 7.01 mmol) suspended in
Toluene (20 mL) was sealed and heated in Biotage Initiator at 150 °C for 7 min. After cooling the solvent was filtered off and the residue was triturated in cyclohexane (ca 100 ml) and methanol (ca 2 ml) and filtered. The title product (1 .1 g) was obtained after drying at 40°C and 0 mmHg for 2 hours in the form of pale green powder. The filtrate was evaporated and triturated once more in the same manner and the title compound (0.35 g) was obtained in the form of pale green powder. The filtrate was then evaporated and purified on Biotage SP1 (Snap Si 50 g; 40 ml/min) in the gradient of EtOAc in Cyclohexane 10% of EtOAc for 2 CV then from 10-40% in 20 CV. The title compound (0.25 g) was obtained in the form of yellow powder; 1H NMR (500 MHz, DMSO-de) δ ppm 12.7 (s, 1 H), 7.44 (m,1 H), 7.2 (m,2H), 3.9 (s,2H).
Intermediate 3: 6-chloro-3-(methylthio)-2H-1 ,4-benzothiazine
Figure imgf000026_0001
To a suspension of potassium hydroxide (0.878 g, 15.65 mmol) and 6-chloro-2H- 1 ,4-benzothiazine-3(4H)-thione (Intermediate 2) (1.35 g, 6.26 mmol) in acetone (40 mL) stirred at room temp was added a solution of methyl iodide (0.391 ml_, 6.26 mmol) portionwise over 15 min and the reaction mixture was stirred at rt for 6 h. The mixture was filtered and evaporated to give a crude product that was purified on Biotage SP1 Snap Si 50g; 40 ml/min.; eluting with EtOAc/Cyclohexane 1/10. The title compound (850 mg) was obtained in the form of dark brown oil; 1H NMR (500 MHz, DMSO-de) δ ppm 7.29-7.33 (m, 1 H), 7.14-7.2 (m,1 H), 7.05-6.99 (m,1 H), 3.2 (s,2H), 2.5 (s,3H); Intermediate 4: ethyl-(6-chloro- -1 ,4-benzothiazin-3(4H)-ylidene)(nitro)ethanoate
Figure imgf000026_0002
To a solution of 6-chloro-3-(methylthio)-2H-1 ,4-benzothiazine (intermediate 3) (1.6 g, 5.92 mmol) and ethyl nitroacetate (7.88 g, 59.2 mmol) stirred under argon at room temp was added a solution of Triton-B (2.96 ml, 6.51 mmol) in one go. The reaction mixture was stirred at 50 °C for 35 hours then at rt for 48 hours. Then it was quenched with water (100ml), extracted with DCM (4x75 ml). Organic layers were combined and washed with ΝθΗΟΟ (sat. sol.) (3x100 ml) and brine. Then they were passed through an 1ST Phase separator and evaporated. The resulting crude oil was purified on Biotage SP1 Snap Si 100 g; 40 ml/min; in a gradient of EtOAc in
Cyclohexane 0% of EtOAc for 1 CV and then from 0-30% in 25 CV to give the title compound as a mixture of E/Z isomers (0.749 g); m/z: 315 [M+1 ].
Intermediate 5: ethyl 8-chloro-4H-[1 ,2,3ltriazolo[5,1-cl[1 ,4lbenzothiazine-3- carboxylate
Figure imgf000027_0001
To a suspension of ethyl (6-chloro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (Intermediate 4) (200 mg, 0.540 mmol) in acetic acid (7 mL) was added zinc (212 mg, 3.24 mmol) portionwise with cooling under argon. The resulting suspension was stirred at rt for 25 min. Then a solution of isoamyl nitrite (0.109 mL, 0.810 mmol) was added in one charge followed by trichloroacetic acid (52.9 mg, 0.324 mmol). The reaction mixture was stirred at rt for 2h. Then it was quenched with water, diluted with brine and extracted with DCM. DCM layer was washed with NaHC03 (sat sol) until evolution of carbon dioxide ceased. The organic phase was dried over sodium sulphate and evaporated. The resulting residue was purified on Biotage SP1 Snap Si 25 g 25 ml/min using a gradient of EtOAc in Cyclohexane 1 % for 2CV then from 1 -30% in 20 CV to give the title compound (104 mg); m/z: 296 [M+1 ]; 1H NMR (500 MHz, CDCI3) 5~/ppm 8.23 (s, 1 H), 7.4-7.42 (m, 1 H), 7.33-7.34 (m,1 H), 4.48-4.49 (q,2H) 4.44 (s,2H), 1.45-1.48 (t,3H).
Intermediate 6: ethyl 8-chloro-1-methyl-4H-imidazor5,1-ciri ,4lbenzothiazine-3- carboxylate
Figure imgf000027_0002
To a solution of ethyl (2E/Z)-(6-chloro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (Intermediate 4) (400 mg, 1.017 mmol) in acetic acid (10 mL) stirred under argon at 0°C was added solid zinc (465 mg, 7.12 mmol) portionwise during 15 min. The reaction mixture was allowed to warm to rt and stirred for 0.5 h. Then triethyl orthoacetate (0.375 mL, 2.033 mmol) was added dropwise during 5 min. The reaction mixture was stirred at rt for 22h then triethyl orthoacetate (0.094 mL, 0.508 mmol) was added and reaction was stirred for another 2h. Then it was quenched with water (50ml) and extracted with DCM (5x50ml). The organic layer was washed with NaHCC>3(sat.sol.) until evolution of CO2 ceased. Then it was dried over sodium sulphate/magnesium sulphate and passed through an 1ST Phase separator and evaporated to give a crude oil (387 mg). This oil was purified on Biotage SP1 Snap Si 25g; 25 ml/min; first in the gradient of 2N NH3/MeOH in DCM 0-7% in 20 CV and then in the gradient of EtOAc in Hexane 12% of EtOAc for 2 CV then from 12-100% in 15CV then 100% of EtOAc for 3 CV to give the title compound (94 mg); m/z: 341.2 [M+1]. Intermediate 7: 8-chloro-1 -methyl-4H-imidazor5,1-ciri ,4lbenzothiazine-3-carboxylate
Figure imgf000028_0001
To a suspension of ethyl 8-chloro-1 -methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3- carboxylate (intermediate 6) 5,5-dioxide (89 mg, 0.261 mmol) in tetrahydrofuran (2 mL) and Water (1 mL) stirred in air at room temp was added solid KOH (29.3 mg, 0.522 mmol) in one charge. The reaction mixture was stirred at rt for 3h. Then it was diluted with water (15ml) acidified to pH 5-6 (strip) by addition of 1 N HCI aq. and extracted with EtOAc (3x30 ml). The organic layer was dried over Na2S0 /MgS0 and evaporated giving a powder which was evaporated again over toluene giving the title compound (64 mg); 1H NMR (500 MHz, DMSO-d6) δ ppm 2.73 (s, 3 H) 5.23 (s, 2 H) 7.78 (dd, J=8.54, 1 .53 Hz, 1 H) 8.04 - 8.10 (m, 2 H)
Intermediate 8: 4-Chloro-5-fluoro-2-nitrophenol
A solution of 4-chloro-3-fluorophenol (4.937 g, 33.7 mmol) in DCM (99 mL) was cooled to 0 °C. To the stirred was added concentrated sulfuric acid (2.81 mL, 50.5 mmol). To this mixture concentrated nitric acid (2.55 mL, 37.1 mmol) was carefully added dropwise to maintain the temperature below 5 °C. The reaction was stirred for 1 hour at 0 °C. Reaction mixture was quenched with water (300 mL). The layers were separated and the aqueous layer was extracted with DCM (2 x 50 mL). The combined organics were washed with sat NaHC03 (100 mL), brine (100 mL) and dried over anhydrous Na2S04. The solvent was removed under vacuum and the crude product was added to a silica gel column and eluted with DCM to obtain the title compound as yellowish solid (3.58 g); m/z (ES): 189.96 [M+H]"; 1H NMR (500 MHz, DMSO-d6) δ ppm: 7.12 (d, J=10.68 Hz, 1 H), 8.22 (d, J=7.93 Hz, 1 H), 1 1.73 (br. s., 1 H).
Intermediate 9: Ethyl [(4-chloro-5-fluoro-2-nitrophenyl)oxylacetate
Figure imgf000029_0001
To a solution of 4-chloro-5-fluoro-2-nitrophenol (Intermediate 8, 7.38 g, 38.5 mmol) in acetone (40 mL) under argon atmosphere, potassium carbonate (5.32 g, 38.5 mmol) was added. After 10 minutes an orange solid was formed. Mixture was diluted with additional acetone (25 mL) in order to facilitate stirring and then ethyl bromoacetate (4.29 mL, 38.5 mmol) was added. Mixture was heated at 70 °C for 3 hours. A new portion of potassium carbonate (1 .597 g, 1 1 .56 mmol) and (2-bromoethyl)benzene (0.289 mL, 2.133 mmol) were added, and heating was continued for additional 2 hours. Reaction mixture was concentrated, diluted with DCM (100 mL), washed with sat NaHCOs (3 x 100 mL), brine (100 mL) and dried over Na2S04. The solvent was removed under vacuum to obtain the title compound as orange solid (10.7 g); m/z (ES): 278.07 [M+H]+; 1H NMR (300 MHz, CHLOROFORM-d) δ ppm: 1 .31 (t, J=7.16 Hz, 3 H), 4.29 (q, J=7.03 Hz, 2 H), 4.77 (s, 2 H), 6.81 (d, J=9.80 Hz, 1 H), 8.06 (d, J=7.54 Hz, 1 H).
Intermediate 10: 6-Chloro-7-fluoro-2H-1 ,4-benzoxazin-3(4/-/)-one
Figure imgf000029_0002
To a suspension of ethyl [(4-chloro-5-fluoro-2-nitrophenyl)oxy]acetate (Intermediate 9, 10.7 g, 38.5 mmol) in a mixture of methanol (78 mL) and water (78 mL) stirred with a mechanical stirrer were added ammonium chloride (20.62 g, 385 mmol) and iron (12.91 g, 231 mmol). The reaction mixture was stirred at 80 °C for 1 day. Hot reaction mixture was filtered through celite and the filter cake was washed with hot MeOH (3 x 50 mL). The organic solvent was removed under vacuum and the resulting aqueous residue was extracted with EtOAc (3 χ 100 mL), and dried over Na2S04. The solvent was removed under vacuum to obtain the title compound (3.74 g). The filter cake contained significant amounts of the desired compound and therefore it was transfered to a round bottom flask and triturated with EtOAc (150 mL). The mixture was heated for 1 hour at 60 °C, and filtered hot through a Buchner funnel to receive a colourless solution. The solvent was evaporated to yield the title compound (2.3 g); m/z (ES): 202.1 1 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 4.63 (s, 2 H), 6.98 (d, J=7.32 Hz, 1 H), 7.15 (d, J=10.07 Hz, 1 H), 10.81 (br. s., 1 H). Intermediate 1 1 : 6-Chloro-7-fluoro-2/-/-1 ,4-benzoxazine-3(4/-/)-thione
Figure imgf000030_0001
To a suspension of 6-chloro-7-fluoro-2/-/-1 ,4-benzoxazin-3(4/-/)-one (intermediate 10, 6.03 g, 29.9 mmol) in dry toluene (100 mL), Lawesson's reagent (7.26 g, 17.95 mmol) was added. The mixture was heated at 100 °C for 3 hours. Reaction mixture was left at room temperature for 2 hours, filtered, and the solid washed with Et20 (3 x 25 mL). The mother liquor was evaporated and toluene (100 mL) was added to the residue (13.9 g). This mixture was treated in an ultrasonic bath for 30-60 minutes, the precipitate filtered off and washed with cold toluene (2 x 15 mL) to yield the title compound (1 .92 g). The mother liquor was concentrated to about 80 mL. During this process a yellowish precipitate was formed. The precipitate was again treated as above [ultrasonic bath for 30-60 minutes, filtration, and washing with cold toluene (2 x 15 L)] to give another portion of the title compound (2.09 g, 31 .8 % yield). The second mother liquor was evaporated to dryness and Et20 (100 mL) was added to the residue. This mixture was treated in an ultrasonic bath for 30-60 minutes; the beige precipitate was filtered off and washed with Et20 (2 x 10 mL) to yield an unknown sideproduct (about 6 g). After evaporation of the mother liquor (Et20) the raw product (3.9 g) was purified on a FlashMaster I I Instrument (LC-Si cartridge 50 g) using n-hexane as eluent to obtain the title compound (1 .6 g); m/z (ES): 216.06 [M- H]-; 1H NMR (500 MHz, DMSO-d6) δ ppm: 4.89 (s, 2 H), 7.19 (d, J=7.63 Hz, 1 H), 7.22 (d, J=10.07 Hz, 1 H), 12.83 (br. s., 1 H).
Intermediate 12: 6-Chloro-7-fluor -3-(methylthio)-2/-/-1 ,4-benzoxazine
Figure imgf000030_0002
To a solution of 6-chloro-7-fluoro-2H-1 ,4-benzoxazine-3(4H)-thione (intermediate 1 1 , 5.577 g, 25.6 mmol) in acetone (156 mL) at room temperature was added potassium hydroxide (2.156 g, 38.4 mmol) followed by iodomethane (2.403 mL, 38.4 mmol). The mixture was stirred at room temperature for 1 hour. Reaction mixture was concentrated, diluted with DCM (100 mL), washed with water (3 x 50 mL), brine (2 χ 50 mL), and dried over IS^SC^, The solvent was removed under vacuum to obtain the title compound as yellow solid (5.49 g); m/z (ES): 232.10 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 2.50 (s, 3 H), 4.71 (s, 2 H), 7.10 (d, J=10.07 Hz, 1 H), 7.40 (d, J=7.93 Hz, 1 H).
Intermediate 13: Ethyl (2E/ZH6-chloro-7-fluoro-2H-1 ,4-benzoxazin-3(4HV
ylidene)(nitro)ethanoate
Figure imgf000031_0001
To a suspension of 6-chloro-7-fluoro-3-(methylthio)-2H-1 ,4-benzoxazine
(intermediate 12, 1 g, 4.32 mmol) in ethyl nitroacetate (4.83 mL, 43.2 mmol), triton-B (1 .962 mL, 4.32 mmol) was added. Reaction mixture was stirred at 50 °C for 20 hours. Reaction mixture was cooled to room temperature, quenched with water (50 mL) and extracted with DCM (3 * 75 mL). Combined organic extracts were washed with sat K2C03 (2 100 mL) and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a FlashMaster II Instrument (LC-Si cartridge 20 g) eluting with an n-hexane/EtOAc gradient starting from 100/0 v/v for 10 minutes going to 0/100 v/v in the next 1 10 minutes, to give the title compound as mixture of geometric isomers (0.108 g); m/z (ES): 315.12 [M-H]"
Intermediate 14: Ethyl 8-chloro-7-fluoro-1-methyl-4H-imidazor5,1-ciri ,4lbenzoxazine-
3-carboxylate
Figure imgf000031_0002
To a solution of ethyl (2£/Z)-(6-chloro-7-fluoro-2H-1 ,4-benzoxazin-3(4H)- ylidene)(nitro)ethanoate (intermediate 13, 189 mg, 0.597 mmol) in acetic acid (8 mL), stirred under argon at 10 °C, solid zinc (273 mg, 4.18 mmol) was added portionwise during 15 minutes. The reaction mixture was allowed to warm to room temperature and stirred for 30 minutes. The reaction mixture was filtered and the filter cake washed with DCM (3 x 10 mL). The organic solvent was removed in vacuo, and triethyl orthoacetate (1.566 mL, 8.49 mmol) and p-toluenesulfonic acid monohydrate (7.83 mg, 0.041 mmol) were added. The suspension was heated at 90 °C for 2 hours. Reaction mixture was diluted with EtOAc (30 mL), washed with sat NaHC03 (50 mL), brine (50 mL), and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a FlashMaster II Instrument (LC-Si cartridge 5 g) eluting with an n-hexane/EtOAc gradient starting from 100/0 v/v going to 25/75 v/v in the 60 minutes to obtain the title compound as yellowish solid (35 mg); m/z (ES): 31 1.13 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm: 1.39 (t, J=7.02 Hz, 3 H), 2.74 (s, 3 H), 4.39 (q, J=7.02 Hz, 2 H), 5.43 (s, 2 H), 6.99 (d, J=8.85 Hz, 1 H), 7.58 (d, J=7.02 Hz, 1 H).
Intermediate 15: Ethyl 8-chloro-7-fluoro-4/-/-ri,2,3ltriazolor5,1-cin,4lbenzoxazine-3- carboxylate
Figure imgf000032_0001
To a solution of ethyl (2£/Z)-(6-chloro-7-fluoro-2H-1 ,4-benzoxazin-3(4H)- ylidene)(nitro)ethanoate (intermediate 13, 44 mg, 0.139 mmol) in acetic acid (1.5 mL), stirred under argon at 10 °C, solid zinc (54.5 mg, 0.834 mmol) was added portionwise during 15 minutes. The reaction mixture was allowed to warm to room temperature and stirred for 1 hour. Then, isoamyl nitrite (0.026 mL, 0.195 mmol) was added followed by trichloroacetic acid (1 1.35 mg, 0.069 mmol) and the mixture was stirred at room temperature over night. Reaction mixture was quenched with water (25 mL) and extracted with DCM (3 x 15 mL). Combined organic extracts were washed with sat NaHC03 (3 x 15 mL), brine (25 mL), and dried over Na2S04. The solvent was removed under vacuum, the crude product was purified on a
FlashMaster II Instrument (LC-Si cartridge 1 g) eluting with an n-hexane/EtOAc gradient starting from 100/0 v/v for 3 minutes going to 25/75 v/v in the next 27 minutes to obtain the title compound (27 mg); m/z (ES): 298.12 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm: 1.43 (t, J=7.17 Hz, 3 H), 4.45 (q, J=7.12 Hz, 2 H), 5.65 (s, 2 H), 6.96 (d, J=9.16 Hz, 1 H), 8.15 (d, J=7.02 Hz, 1 H).
Intermediate 16: 1 ,3,4,5-Tetrahydro-2H-1-benzazepin-2-one
Figure imgf000032_0002
To a suspension of 3,4-dihydro-1 (2H)-naphthalenone (91 μί, 0.684 mmol) and sodium azide (55.6 mg, 0.855 mmol) in chloroform (700 μί) was stirred at 40 °C, concentrated sulphuric acid (182 μί, 3.42 mmol) was gradually added over a period of 1 hour. The reaction mixture was cooled to room temperature, diluted with water (15 mL) and extracted with DCM (3 x 15 mL). Combined organic extracts were washed with sat NaHC03 (25 mL), brine (25 mL), and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a
FlashMaster II Instrument (LC-Si cartridge 2 g) eluting with an n-hexane/EtOAc gradient starting from 95/5 v/v for 5 minutes going to 50/50 v/v in the next 30 minutes to obtain the title compound as white solid (100 mg); m/z (ES): 162.02 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 2.04 - 2.17 (m, 4 H), 2.67 (t, J=6.87 Hz, 2 H), 6.95 (d, J=7.63 Hz, 1 H), 7.03 - 7.10 (m, 1 H), 7.18 - 7.27 (m, 2 H), 9.52 (br. s., 1 H).
Intermediate 17: 1 ,3,4,5-Tetrahydro- -1-benzazepine-2-thione
Figure imgf000033_0001
To a suspension of 1 ,3,4,5-tetrahydro-2H-1-benzazepin-2-one (intermediate 16, 3.63 g, 22.52 mmol) in dry toluene (70 mL), Lawesson's reagent (5.46 g, 13.51 mmol) was added. The mixture was heated at 100 °C for 2 hours. Reaction mixture was left at room temperature for 2 hours, filtered, and the solid washed with DCM (3 x 10 mL). The mother liquor was evaporated, the raw product was purified several times on a FlashMaster II instrument using appropriate mixture of EtOAc/cyclohexane (0-25%) as eluent to obtain the title compound as white solid (3.28 g); m/z (ES): 178.16
[M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 2.19 (quin, J=7.17 Hz, 2 H), 2.62 (dt, J=14.34, 7.17 Hz, 4 H), 7.02 - 7.09 (m, 1 H), 7.15 - 7.24 (m, 1 H), 7.25 - 7.32 (m, 2 H), 1 1.76 (br. s., 1 H).
Intermediate 18: 2-(Methylthio)-4,5-dihvdro-3H-1-benzazepine
—
Figure imgf000033_0002
To a solution of 1 ,3,4,5-tetrahydro-2H-1-benzazepine-2-thione (intermediate 17, 3.275 g, 18.48 mmol) in acetone (156 mL) at room temperature, potassium hydroxide (2.59 g, 46.2 mmol) was added followed by iodomethane (1.271 mL, 20.32 mmol). The mixture was stirred at room temperature for 3 hours. Reaction mixture was diluted with water (200 mL), extracted with DCM (3 x 50 mL), washed with brine (150 mL) and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a FlashMaster II Instrument (LC-Si cartridge 20 g) eluting with a cyclohexane/EtOAc gradient starting from 100/0 v/v for 70 minutes going to 95/5 v/v in the next 40 minutes to obtain the title compound as colourless oil (3.21 g); m/z (ES): 192.15 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 2.23 - 2.38 (m, 4 H), 2.50 (s, 3 H), 2.51 - 2.55 (m, 2 H), 7.02 (d, J=7.63 Hz, 1 H), 7.05 - 7.14 (m, 1 H), 7.21 - 7.38 (m, 2 H). Intermediate 19: Ethyl 2-nitro(1 ,3A5-tetrahydro-2H-1-benzazepin-2- ylidene)ethanoate
Figure imgf000034_0001
To a solution of 2-(methylthio)-4,5-dihydro-3H-1-benzazepine (intermediate 18, 1 .0 g, 5.23 mmol) in ethyl nitroacetate (2.92 mL, 26.1 mmol) at room temperature DBU (0.867 mL, 5.75 mmol) was added and the mixture was left to stir at 40 °C for 5 days. Reaction mixture was quenched with water (100 mL) and extracted with DCM (3 * 30 mL). Combined organic extracts were washed with sat K2C03 (3 * 50 mL), brine (50 mL), and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a FlashMaster II Instrument (LC-Si cartridge 20 g) eluting with an n-hexane/EtOAc gradient starting from 100/0 v/v for 5 minutes going to 75/25 v/v in the next 1 15 minutes to obtain the title compound as unknown single geometric isomer (1.13 g); m/z (ES): 275.15 [M+H]"; 1H NMR (500 MHz, DMSO-d6) δ ppm: 1.26 (t, J=7.17 Hz, 3 H), 2.17 - 2.28 (m, 4 H), 2.69 (t, J=6.71 Hz, 2 H), 4.26 (q, J=7.22 Hz, 2 H), 7.23 - 7.30 (m, 1 H), 7.31 - 7.38 (m, 3 H), 1 1.31 (br. s., 1 H).
Intermediate 20: Ethyl 5,6-dihvdro-4H-ri .2,31triazolori .5-aimbenzazepine-3- carboxylate
Figure imgf000034_0002
To a solution of ethyl 2-nitro(1 ,3,4,5-tetrahydro-2H-1-benzazepin-2-ylidene)ethanoate (intermediate 19, 150 mg, 0.543 mmol) in glacial acetic acid (6 mL), stirred under argon at 10 °C, solid zinc (213 mg, 3.26 mmol) was added portionwise during 15 minutes. The reaction mixture was allowed to warm to room temperature and stirred at this temperature for 1 hour. Then isoamyl nitrite (0.102 mL, 0.760 mmol) was added followed by trichloroacetic acid (44.4 mg, 0.271 mmol) and the mixture was stirred at room temperature over night. Reaction mixture was quenched with water (50 mL) and extracted with DCM (3 x 15 ml_). Combined organic extracts were washed with sat NaHC03 (3 x 30 mL), brine (50 mL), and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a FlashMaster II Instrument (LC-Si cartridge 2 g) eluting with a cyclohexane/EtOAc gradient starting from 100/0 v/v going to 80/20 v/v in the 50 minutes to obtain the title compound (43.9 mg); m/z (ES): 258.19 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 1 .34 (t, J=7.17 Hz, 3 H), 2.23 (quin, J=7.M Hz, 2 H), 2.55 (t, J=7.02 Hz, 2 H), 2.99 (t, J=7.32 Hz, 2 H), 4.36 (q, J=7.02 Hz, 2 H), 7.49 - 7.59 (m, 3 H), 7.69 - 7.77 (m, 1 H).
Intermediate 21 : Ethyl 1-methyl-5,6-dihvdro-4H-imidazo[1 ,5-al[1lbenzazepine-3- carboxylate
Figure imgf000035_0001
To a solution of ethyl 2-nitro(1 ,3,4,5-tetrahydro-2H-1-benzazepin-2-ylidene)ethanoate (intermediate 19, 0.140 g, 0.507 mmol) in acetic acid (4.30 mL), stirred under argon at 10 °C, solid zinc (0.232 g, 3.55 mmol) was added portionwise during 15 minutes. The reaction mixture was allowed to warm to room temperature and stirred for 1 hour. The reaction mixture was filtered and the filter cake was washed with DCM (3 χ 10 mL). The organic solvent was removed in vacuo, and triethyl orthoacetate (1 .326 mL, 7.20 mmol) and p-toluenesulfonic acid monohydrate (6.75 mg, 0.035 mmol) were added. The suspension was heated at 90 °C over night. Reaction mixture was diluted with EtOAc (30 mL), washed with sat NaHC03 (50 mL), brine (50 mL), and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a FlashMaster II Instrument (LC-Si cartridge 5 g) eluting with a
DCM/(DCM/MeOH 30/1 ) gradient starting from 100/0 v/v going to 0/100 v/v in the 60 minutes to obtain the title compound as brownish oil (69.4 mg); m/z (ES): 271 .17 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 1.28 (t, J=7.02 Hz, 3 H), 1.94 - 2.09 (m, 3 H), 2.19 - 2.29 (m, 1 H), 2.33 (s, 3 H), 2.67 (dd, J=13.28, 5.65 Hz, 1 H), 3.48 - 3.58 (m, 1 H), 4.16 - 4.30 (m, 2 H), 7.39 - 7.49 (m, 3 H), 7.49 - 7.55 (m, 1 H).
Intermediate 22: 2H-1 ,4-benzothiazine-3(4H)-thione
Figure imgf000036_0001
To a solution of 2H-1 ,4-benzothiazin-3(4H)-one (400 mg, 2.421 mmol) in dry toluene (4 mL) in a glass tube was added Lawesson's reagent (881 mg, 2.179 mmol). The tube washeated in a microwave reactor at 100 °C for 5 minutes. Thus solvent was removed under reduced pressure and the residue was dissolved in DCM/MeOH = 99:1. The resulting precipitate was filtered and the filtrate was evaporated. The residue was purified via Biotage SP1 purification system using Normal phase Silica SNAP 2x100 g column in a gradient of ethyl acetate in cyclohexane (0 % in 1 CV, 0- 22 % in 14 CV, 22 % in 5 CV) to obtain the title compound (5 g); UPLC/MS Rt = 4.27; m/z (ES): 182.39 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 3.87 (s, 2 H) 6.93 (d, 1 H) 7.08 (t, 1 H) 7.19 (t, 1 H) 7.33 (d, 1 H) 9.68 (s, 1 H).
Intermediate 23: 3-(methylthio)-2 -1 ,4-benzothiazine
Figure imgf000036_0002
To a solution of 2H-1 ,4-benzothiazine-3(4H)-thione (Intermediate 22; 5 g, 27.6 mmol) in acetone (165 mL) at rt was added potassium hydroxide (3.87 g, 69.0 mmol) followed by half amount of methyl iodide (1.29 mL). The mixture was stirred at rt for 15 min, then the rest of methyl iodide (1.29 mL) was added and the mixture was stirred for 1 h at rt. The solvent was removed under reduced pressure and the residue was purified via Biotage SP1 purification system using Normal phase Silica SNAP 100 g column in a gradient of ethyl acetate in cyclohexane (8 % in 1 CV, 8-66 % in 15 CV, 66 % in 2 CV) to obtain the title compound (4.7 g); HPLC/MS Rt = 7.19; m/z (ES): 196.81 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 2.56 (s, 3 H) 3.24 (s, 2 H) 7.05 (t, 1 H) 7.18 (t, 1 H) 7.24 (d, 1 H) 7.30 (d, J=7.93 Hz, 1 H).
Intermediate 24: ethyl (2Z)-2H-1 -benzothiazin-3(4H)-ylidene(nitro)ethanoate
Figure imgf000036_0003
To a solution of 3-(methylthio)-2H-1 ,4-benzothiazine (Intermediate 23; 4.4 g, 22.53 mmol) in ethyl nitroacetate (10.00 ml, 90 mmol) at rt and under argon was added Triton-B 40 % in MeOH (10.24 ml, 22.53 mmol) and the mixture was heated to 50 °C and stirred at that temperature for 5 h. The mixture was then left at rt for 3 days. The reaction mixture was quenched with water (40 mL) and extracted with DCM (3x40 mL). The combined organic phases were washed with K2CO3 aq solution (2x30 mL) and dried using a phase separator filter tube and concentrated under reduced pressure. The residue was purified via Biotage SP1 purification system using Normal phase Silica SNAP 100 g column in the gradient of ethyl acetate in cyclohexane (3 % in 1 CV, 3-22 % in 10 CV, 22-64 % in 10 CV) to give the title compound (1.8 g); m/z (ES): 281 .47 [M+H]+; 1 H NMR (500 MHz, CHLOROFORM-d) δ ppm 1.36 (t, 3 H) 1.38 (t, 3 H) 3.75 (s, 2 H) 3.76 (s, 2 H) 4.34 (q, 2 H) 4.38 (q, 2 H) 7.04 (d, 1 H) 7.07 - 7.17 (m, 3 H) 7.21 (t, 1 H) 7.23 - 7.25 (m, 1 H) 7.30 (d, 1 H) 7.33 (d, 1 H) 1 1.48 (s, 1 H) 1 1.95 (s, 1 H).
Intermediate 25: ethyl 1 -methyl-4H-imidazo 5,1-cl[1 ,41benzothiazine-3-carboxylate
Figure imgf000037_0001
To a solution of ethyl (2Z)-2H-1 ,4-benzothiazin-3(4H)-ylidene(nitro)ethanoate (Intermediate 24; 700 mg, 2.497 mmol) in acetic acid (16 mL) was added zinc (868 mg, 13.27 mmol) and the suspension was stirred at rt for 25 min. Then 1 ,1 ,1- tris(ethyloxy)ethane (0.916 mL, 4.99 mmol) was added and the mixture was stirred at rt overnight. Reaction mixture was quenched with water (20 mL) and extracted with DCM (3x20 mL). Organic phase was washed with a saturated solution of NaHCC>3 (2x20 mL) and water (1x20 mL), then dried over Na2SC> , filtered and evaporated. The residue was purified via Biotage SP1 purification system using Normal phase Silica SNAP 10 g column in the gradient of ethyl acetate in cyclohexane (3 % in 1 CV, 3-9 % in 1 CV, 9-63 % in 12 CV) to give the title compound (145 mg); m/z (ES): 275.51 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) d ppm 1 .42 (t, 3 H) 2.69 (s, 3 H) 4.31 (s, 2 H) 4.42 (q, 2 H) 7.27 - 7.29 (m, 1 H) 7.33 - 7.37 (m, 1 H) 7.51 - 7.57 (m, 2 H).
Intermediate 26: N-(6-chloro-2-pyridinyl)-1-methyl-4H-imidazor5,1- ciri ,41benzothiazine-3-carboxamide
Figure imgf000037_0002
To a solution of 6-chloro-2-pyridinamine (187 mg, 1.458 mmol) in dry 1 ,4-dioxane (2 mL) at rt was added dropwise trimethylaluminium (0.729 mL, 1.458 mmol) under an argon atmosphere. The solution was stirred at rt for 1 hour and then a solution of ethyl 1 -methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 25, 100 mg, 0.365 mmol) in dry 1 ,4-Dioxane (2 mL) was added. The reaction mixture was heated at 95 °C for 1 .5 h. Reaction mixture was cooled at 0 °C and quenched with dioxane/water (20/1 mL) and stirred for 10 min. It was concentrated and dissolved in water (25 mL) and extracted with DCM (4 x 25 ml). The combined organic extracts were washed with 1 N HCI (3x20 ml) and sat NaCI (25 ml), and dried over anhydrous IS^SC^. The solvent was filtered over phase separator filter tube and removed in vacuo to give a crude product. Crude product (low solubility) was triturated with MeOH to obtain the title product (72 mg); m/z (ES): 357.20 [M+H]+; 1 H NMR (500 MHz, DMSO-d6) d ppm 2.68 (s, 3 H) 4.41 (s, 2 H) 7.27 (d, 1 H) 7.37 (t, 1 H) 7.46 (t, 1 H) 7.64 (d, 1 H) 7.85 (d, 1 H) 7.92 (t, 1 H) 8.18 (d, 1 H) 9.61 (s, 1 H).
Intermediate 27: ethyl 4H-[1 ,2,3ltriazolo[5,1-cl[1 ,4lbenzothiazine-3-carboxylate
Figure imgf000038_0001
To a solution of ethyl (2Z)-2H-1 ,4-benzothiazin-3(4H)-ylidene(nitro)ethanoate
(intermediate 24, 1526 mg, 5.44 mmol) in Acetic Acid (36.5 mL) cooled in an ice bath was added zinc (2136 mg, 32.7 mmol) portionwise. The temperature of the mixture was allowed to reach room temperature during next 30 minutes and then isoamyl nitrite (1 .026 mL, 7.62 mmol) was added followed by trichloroacetic acid (445 mg, 2.72 mmol) and the mixture was stirred at rt overnight. Reaction mixture was quenched with water (30 mL) and extracted with DCM (3x30 mL). Organic phase was washed with saturated NaHCC>3 solution (2x30 mL) and water (1 x30 mL), then dried over
Figure imgf000038_0002
filtered and evaporated. Crude was purified via Biotage SP1 purification system using Normal phase Silica SNAP 50 g column in the gradient of ethyl acetate in cyclohexane (5 % in 1 CV, 5-40 % in 10 CV, 40-80 % in 10 CV) to give the title compound (1 .1 g); m/z (ES): 262.1 [M+H]+; 1H NMR (500 MHz,
CHLOROFORM-d) d ppm 1.46 (t, 3 H) 4.44 (s, 2 H) 4.48 (q, 2 H) 7.34 (t, 1 H) 7.39 (t, 1 H) 7.48 (d, 1 H) 8.20 (d, 1 H).
Intermediate 28: ethyl 4H-imidazor5,1-ciri.4lbenzothiazine-3-carboxylate
Figure imgf000039_0001
Potassium tert-butoxide (3.60 g, 32.1 mmol) was added to a solution of 2H-1 ,4- benzothiazin-3(4H)-one (intermediate 24, 5 g, 30.3 mmol) in dry N,N- dimethylformamide (DMF) (90 mL) which was cooled to 5 °C under argon and stirred for 10 minutes. Diethyl chlorophosphate (7.83 mL, 54.5 mmol) was added and stirring continued for a further 5 minutes. Ethyl isocyanoacetate (4.98 mL, 43.3 mmol) in dry N,N-dimethylformamide (45 mL) and potassium tert-butoxide (4.75 g, 42.4 mmol) were added and the mixture was stirred at 5 °C for 15 minutes and then at room temperature overnight. The reaction was acidified with 5 mL of acetic acid, diluted with 70 mL of water and poured slowly into 400 mL of vigorously stirred ice- water. After 0.5 h, the solid was collected, washed with water, air dried and precipitated with Cy : EtOAc = 10 : 1 to give the title compound (1.3 g); m/z (ES): 261.20 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 1.31 (t, J=7.17 Hz, 3 H) 4.28 (q, 2 H) 4.44 (s, 2 H) 7.32 (t, 1 H) 7.40 (t, 1 H) 7.56 (dd, J=7.78, 1.37 Hz, 1 H) 7.90 (d,1 H) 8.51 (s, 1 H).
Intermediate 29: ethyl 1 -bromo-4H-imidazo[5,1-cl[1 ,4lbenzothiazine-3-carboxylate
Figure imgf000039_0002
In a 250 mL three-necked flask ethyl 4H-imidazo[5,1 -c][1 ,4]benzothiazine-3- carboxylate (intermediate 28, 870 mg, 3.34 mmol) was dissolved in dry
tetrachloromethane (80 mL) under argon atmosphere. After stirring was commenced, the resulting suspension was treated with NBS (654 mg, 3.68 mmol) and AIBN (165 mg, 1.003 mmol) in portions and the mixture was heated at 60 °C for 5 h. The reaction was quenched with sat NaHCC>3 (60 mL) and extracted with EtOAc (3x60 mL). Combined organic layers were washed with brine, dried over
Figure imgf000039_0003
filtered through a phase separator filter tube and the solvent removed in vacuo. The residue was purified via Biotage SP1 purification system using Normal phase Silica SNAP 25 g column in the gradient of EtOAc in Cy (8 % in 1 CV, 8-66 % in 18 CV) to obtain the title compound (539 mg); m/z (ES): 341.20 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 1.31 (t, 3 H) 4.29 (q, 2 H) 4.36 (s, 2 H) 7.40 (t, 1 H) 7.48 (t, 1 H)7.67 (d, 1 H) 8.12 (d, 1 H). Intermediate 30: ethyl 1 -(trifluoromethyl)-4H-imidazo[5,1-cl[1 ,4lbenzothiazine-3- carboxylate
Figure imgf000040_0001
To a solution of ethyl 1-bromo-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 29, 40 mg, 0.1 18 mmol) in dry Ν,Ν-Dimethylformamide (DMF) (300 μΙ_) and N-Methyl-2-pyrrolidone (NMP) (300 μΙ_) mixed in microwave tube were added trimethyl(trifluoromethyl)silane (27.6 μί, 0.177 mmol), copper(l) iodide (35.9 mg, 0.189 mmol) and potassium fluoride (10.28 mg, 0.177 mmol). Reaction was performed in microwave reactor at 90 °C for 10 min. In reaction mixture was added EtOAc (10 mL) and it was washed with water (3x10 mL) and brine (10 mL). Organics were dried over anhydrous IS^SC^. Solvent was filtered over phase separator filter tube and removed in vacuo to give a crude product. Crude product was purified via Biotage SP1 purification system using Normal phase Silica SNAP 10 g column in the gradient of EtOAc in Cy (0 % in 2 CV, 0-55 % in 10 CV, 55-85 % in 10 CV) to give the title compound (12 mg); m/z (ES): 329.07 [M+H]+; 1 H NMR (600 MHz, DMSO-d6) δ ppm 1.23 - 1 .26 (m, 4 H) 4.25 (q, 2 H) 4.32 (s, 2 H) 7.35 - 7.39 (m, 1 H) 7.41 - 7.47 (m, 1 H) 7.56 - 7.59 (m, 1 H) 7.62 -7.65 (m, 1 H).
Intermediate 31 : ethyl 1 -cvclopropyl-4H-imidazor5,1-ciri,4lbenzothiazine-3- carboxylate
Figure imgf000040_0002
To a solution of ethyl 1-bromo-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 29, 150 mg, 0.442 mmol) in dry 1 ,2-dichloroethane (3.5 mL) under argon in a microwave tube were added cyclopropylboronic acid (76 mg, 0.884 mmol), tetrakis(triphenylphosphine)palladium(0) (51.1 mg, 0.044 mmol) and potassium phosphate tribasic (1.548 mL, 1.548 mmol). The reaction was heated in a microwave reactor at 130 degC for 30 minutes then at 140 degC for 30 minutes. The reaction was quenched with water and extracted with EtOAc (3x30 mL). The combined organic layers were washed with brine, then dried (anhydrous Na2SC> ), filtered and concentrated. Crude was purified via Biotage SP1 purification system using Normal phase Silica SNAP 25 g column in the gradient of ethyl acetate in cyclohexane (5 % in 1 CV, 5-30 % in 8 CV, 30-60 % in 8 CV) to obtain product = N 12201 -25-1 (45 mg). Compound N12201 -25-1 was decomposed after one day (1 H NMR: N12201 -25-1 ). It was recrystallized in Cy and EtOAc and checked by UPLC-MS/UV (N 12201 -25-3). By UPLC-MS/UV compound N12201 -25-3 has 3 major peaks. It was purified via Biotage SP1 purification system using Normal phase Silica SNAP 10 g column in the gradient of ethyl acetate in cyclohexane (5 % in 1 CV, 5-30 % in 10 CV, 30-60 % in 8 CV) to obtain the title compound (22 mg); m/z (ES): 301.30 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 1 .02 - 1.07 (m, 4 H) 1.28 (t, J=7.02 Hz, 3 H) 2.09 - 2.16 (m, 1 H) 4.24 (q, J=7.02 Hz, 2 H) 4.31 (s, 2 H) 7.31 - 7.37(m, 1 H) 7.42 - 7.47 (m, 1 H) 7.61 - 7.65 (m, 1 H) 8.00 - 8.07 (m, 1 H).
Intermediate 32: ethyl [(5-fluoro- -nitrophenyl)oxylacetate
Figure imgf000041_0001
To a solution of 5-fluoro-2-nitrophenol (5 g, 31 .8 mmol) in acetone (25 mL) under Argon was added potassium carbonate (4.40 g, 31 .8 mmol). After 5 min an orange solid formed. Mixture was diluted with additional 30 mL of acetone to facilitate stirring and then ethyl bromoacetate (3.54 mL, 31.8 mmol) was added. Mixture was heated at 70 °C for 2.5 h. Reaction mixture was evaporated. Crude was dissolved with EtOAc and washed with a saturated solution of NaHCC^. Organics dried over Na2SC> . The solvent was filtered through a phase separator filter tube and removed in vacuo to give the title compound (7.6 g); m/z (ES): 244.20 [M+H]+; 1 H NMR (500 MHz, DMSO-d6) δ ppm 1.21 (t, J=7.02 Hz, 3 H) 4.17 (q, J=7.02 Hz, 2 H) 5.06 (s, 2 H) 7.03 (ddd, J=9.00, 7.78, 2.75 Hz, 1 H) 7.34 (dd, J=10.99, 2.75 Hz, 1 H) 8.03 (dd, J=8.85, 6.10 Hz, 1 H).
Intermediate 33: 7-fluoro-2H-1 ,4-benzoxazin-3(4H)-one
Figure imgf000041_0002
To a suspension of ethyl [(5-fluoro-2-nitrophenyl)oxy]acetate (intermediate 32, 7.6 g, 31 .3 mmol) in methanol (40 mL) / water (40 mL) was added ammonium chloride (16.72 g, 313 mmol) followed by iron (10.47 g, 188 mmol) powder and the mixture was heated at 80 °C for 2hr. The reaction mixture was filtered through celite, the filtrate was evaporated under vacuo, and the residue was extracted with EtOAc (4x40 mL). The organic phases were washed with brine and dried over IS^SC^. The mixture was filtered through a phase separator filter tube and the mixture was removed in vacuo to give the title product; m/z (ES): 168.10 [M+H]+; 1 H NMR (500 MHz, DMSO-d6) δ ppm 4.59 (s, 2 H) 6.77 - 6.83 (m, 1 H) 6.85 - 6.91 (m, 2 H) 10.68 - 10.74 (m, 1 H).
Figure imgf000042_0001
To a solution of 2H-1 ,4-benzothiazin-3(4H)-one (intermediate 33, 3.1 g, 18.55 mmol) in dry Toluene (15 mL) in a glass tube was added Lawesson's reagent (4.50 g, 1 1 .13 mmol, 0.6 eqv) and the mixture reacted in a microwave reactor at 100 °C for 5 minutes). Cyclohexane was added, and the resulting solid was filtered. The solid was dissolved in Et^O and the solvent removed not quite to dryness. This concentrated mixture was purified using a Biotage SP1 purification system using Normal phase Silica SNAP 4x50 g (because of low solubility) column in the gradient of ethyl acetate in cyclohexane (8 % in 1 CV, 8-30 % in 10 CV, 30-66 % in 10 CV) to give the title compound (2 g); m/z (ES): 184.10 [M+H]+; 1H NMR (600 MHz, DMSO-d6) δ ppm 4.86 (s, 2 H) 6.84 - 6.88 (m, 1 H) 6.92 - 6.97 (m, 1 H) 7.07 - 7.1 1 (m, 1 H) 12.77 (s, 1 H).
Intermediate 35: 7-fluoro-3-(methylthio)-2H-1 ,4-benzoxazine
Figure imgf000042_0002
To a solution of 7-fluoro-2H-1 ,4-benzoxazine-3(4H)-thione (intermediate 34, 2 g, 10.92 mmol) in acetone (70 mL) at rt was added potassium hydroxide (1.225 g, 21 .83 mmol) followed by methyl iodide (0.5 mL) and the mixture was stirred at rt for 15 min. Further methyl iodide (0.251 mL) was added and the mixture was stirred for 40 min at rt. Reaction mixture was quenched with NaHCC>3 sat sol (40 mL) and extracted with DCM (4x30 mL). Combined organic layers were washed with brine (30 mL), dried over Na2SC>4, filtered through a phase separator to remove the solid and the solvent was removed under reduced pressure to give the title compound (2.1 g); m/z (ES): 198.20 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 2.50 (s, 3 H) 4.66 (s, 2 H) 6.79 - 6.86 (m, 2 H) 7.21 - 7.27 (m, 1 H). Intermediate 36: ethyl (2Z)-(7-fluoro-2H-1 ,4-benzoxazin-3(4H)- ylidene)(nitro)ethanoate
Figure imgf000043_0001
To a solution of 7-fluoro-3-(methylthio)-2H-1 ,4-benzoxazine (intermediate 35, 2 g, 10.14 mmol) in ethyl nitroacetate (1 1.26 ml, 101 mmol) at room temperature and under Argon stream, Triton-B (4.61 ml, 10.14 mmol) 40 % in MeOH was added and the mixture was heated to 50 °C and stirred at that temperature overnight. Reaction mixture was quenched with water (40 mL) and extracted with DCM (4x30 mL).
Organics were washed with K2CO3 aq solution (3x30 mL) and brine (30 mL).
Combined organic layers were dried over phase separator filter tube and
concentrated under reduced pressure. Crude was purified via Biotage SP1 purification system using Normal phase Silica SNAP 50 g column in the gradient of ethyl acetate in cyclohexane (3 % in 1 CV, 3-25 % in 10 CV, 25-66 % in 10 CV) to give the title compound (530 mg); m/z (ES): 283.0 [M+H]+; 1H NMR (600 MHz, DMSO-d6) δ ppm 1.26 (t, 3 H) 4.27 (q, 2 H) 5.07 (s, 2 H) 6.88 - 6.96 (m, 1 H) 6.98 - 7.03 (m, 1 H) 7.56 - 7.64 (m, 1 H) 1 1.33 - 1 1.59 (m, 1 H).
Intermediate 37: ethyl 7-fluoro-4H-[1 ,2,3ltriazolo[5,1 -cl[1 ,4lbenzoxazine-3- carboxylate
Figure imgf000043_0002
To a solution of ethyl (2Z)-(7-fluoro-2H-1 ,4-benzoxazin-3(4H)- ylidene)(nitro)ethanoate (300 mg, 1.063 mmol) in acetic acid (12 mL) cooled in an ice bath under Argon atmosphere, zinc (417 mg, 6.38 mmol) was added portionwise. The temperature of the mixture was allowed to reach room temperature during next 30 minutes and then isoamyl nitrite (0.2 mL, 1.488 mmol) was added followed by addition of trichloroacetic acid (87 mg, 0.531 mmol). Reaction was stirred at rt overnight. Reaction mixture was quenched with water (30 mL) and extracted with DCM (3x30 mL). Organic phase was washed with NaHC03 saturated solution (3x20 mL) and water (1 x30 mL), then dried over Na2S04, filtered and evaporated. Crude was purified via Biotage SP1 purification system using Normal phase Silica SNAP 10 g column in the gradient of ethyl acetate in cyclohexane (5 % in 1 CV, 5-40 % in 10 CV, 40-80 % in 10 CV) to give the title compound (240 mg); m/z (ES): 264.3 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 1.34 (t, J=7.02 Hz, 3 H) 4.35 (q, J=7.12 Hz, 2 H) 5.76 (s, 2 H) 7.09 (td, J=8.77, 2.59 Hz, 1 H) 7.23 (dd, J=9.61 , 2.59 Hz, 1 H) 8.08 (dd, J=9.00, 5.65 Hz, 1 H). Intermediate 38: ethyl 7-fluoro-1 -methyl-4H-imidazor5,1-ciri,4lbenzoxazine-3- carboxylate
Figure imgf000044_0001
To a solution of ethyl (2Z)-(7-fluoro-2H-1 ,4-benzoxazin-3(4H)- ylidene)(nitro)ethanoate (intermediate 36, 218 mg, 0.772 mmol) in Acetic Acid (8 mL) cooled in an ice bath under Argon atmosphere, zinc (303 mg, 4.63 mmol) was added portionwise. The temperature of the mixture was allowed to reach room temperature during next 30 minutes and then triethyl orthoacetate (0.356 mL, 1.931 mmol) was added and stirred for 4 h. Reaction mixture was quenched with water (30 mL) and extracted with DCM (3x30 mL). Organic phase was washed with saturated NaHCC>3 solution (3x20 mL) and water (1 x30 mL), then dried over
Figure imgf000044_0002
filtered and evaporated. Crude was purified via Biotage SP1 purification system using Normal phase Silica SNAP 10 g column in the gradient of ethyl acetate in cyclohexane (12 % in 1 CV, 12-100 % in 15 CV, 100 % in 5 CV) to give the title compound (91.3 mg); m/z (ES): 277.30 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 1.29 (t, 3 H) 2.67 (s, 3 H) 4.25 (q, 2 H) 5.44 (s, 2 H) 7.04 - 7.09 (m, 1 H) 7.16 - 7.21 (m, 1 H) 7.81 - 7.86 (m, 1 H).
Figure imgf000044_0003
In a 1000 ml three-neck round bottom flask were combined 300 ml of acetic acid, 4- chloroaniline (20 g, 157 mmol) and potassium thiocyanate (30.5 g, 314 mmol) with continous mechanical stirring. Bromine (12.12 mL, 235 mmol), dissolved in 300 ml of acetic acid, was added dropwise to the reaction mixture (over 30 minutes) whilst keeping the temperature between 18-25 °C throughout the addition using an ice bath). Stirring was continued for an additional 1 h after bromine addition. The mixture was filtered to give a green precipitate which was dried then washed using warm water (2 litres at 35 °C) The washings were basified with 25 % aqueous ammonia solution to pH 1 1-12 (during base addition the mixture was cooled with ice added to the mixture). A white precipitate was formed which was filtered, washed with NH4OH water solution (pH of the solution in range 1 1 -12) and dried in a vacuum oven at 40 °C to obtain the title compound (22 g); m/z (ES): 185.02 [M+H]+; 1H NMR (500 MHz, DMSO-de) δ ppm 7.21 (dd, J=8.54, 2.14 Hz, 1 H) 7.30 (d, J=8.55 Hz, 1 H) 7.59 (s, 2 H) 7.77 (d, J=2.14 Hz, 1 H).
Intermediate 40: 2-amino-5-chlorobenzenethiol
6-Chloro-1 ,3-benzothiazol-2-amine (intermediate 39, 8.7 g, 47.1 mmol) was suspended in 50 % aqueous sodium hydroxide solution (120 mL) and stirred at reflux for 18 hours. Water (50 ml) was added and the mixture was filtered through a Buchner funnel. Adding ice in the mixture and cooling it in the ice bath, the mixture was vigorously stirred and pH of the mixture was adjusted to 6.5 to 7 using glacial acetic acid. Volume of the mixture was increased to 400 ml and the mixture was extracted with diethyl ether (800 ml was used in total), the combined organic layers were washed with wather, dried (IS^SC^) and the solvent was evaporated to obtain the title compound (5.90 g) in a mixture with 2,2'-dithiobis(4-chloroaniline); m/z (ES): 159.78 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) 5ppm 2.98 (br. s., 1 H) 4.13 (br. s., 2 H) 6.65 (d, J=8.70 Hz, 1 H) 7.04 (dd, J=8.54, 2.44 Hz, 1 H) 7.34 (d, J=2.29 Hz, 1 H).
Intermediate 41 : 7-chloro-2H-1 ,4-benzothiazin-3(4H)-one
To a solution of 2-amino-5-chlorobenzenethiol (Intermediate 40, 6 g, 33.8 mmol) in Dimethyl Sulfoxide (DMSO) (150 mL) at room temperature was added sodium ethoxide (2.53 g, 37.2 mmol). After the mixture was stirred for 1 hour, methyl bromoacetate (3.43 mL, 37.2 mmol) was added. The mixture was then heated at 65 °C for 3 hours. Then 350 ml of cold water was added. The resulting precipitate was filtered and was dried in a vacuum oven at 40 °C to give the title compound (3.5 g). the organic substances from the filtrate were extracted with diethyl ether (800 ml in total, several times). The organic layers were washed with the mixture of 1 N HCI (3 x 300 ml). The organic layer was dried over IS^SC^ and the solvent evaporated to give the title compound (3 g); m/z (ES): 199.98 [M+H]+; 1H NMR (500 MHz,
CHLOROFORM-d) Bppm 3.43 (s, 2 H) 6.79 (d, J=8.54 Hz, 1 H) 7.14 (dd, J=8.47, 2.21 Hz, 1 H) 7.32 (d, J=2.14 Hz, 1 H) 8.49 (br. s., 1 H).
Intermediate 42: 7-chloro-2H-1 ,4-benzothiazin-3(4H)-one
Figure imgf000046_0001
7-Chloro-2H-1 ,4-benzothiazin-3(4H)-one (Intermediate 41 , 4 g, 20.03 mmol) was split in two portions (2g) and was put in two 20 ml crucibles for microwave synthesis. In each crucible was put Lawesson's reagent (5.27 g, 13.02 mmol) (2.65 g) and 15 ml of toluene. The reaction mixture was carried out in a microwave reactor at 150 °C for 7 minutes at 400 Watts. The reaction mixture was then suspended in 200 ml of cyclohexane, and the resulting precipitate was filtered using a Buchner funnel. The solid was washed with cyclohexane (400 ml in total) on the Buchner funnel. The precipitate was dried in a vaccum oven at 40 °C, and then suspended in 80 ml of DCM using ultrasound bath. The precipitate was filtered, washed twice with 5 ml of DCM and dried (sodium sulfate) to give (3 g); m/z (ES): 215.8 [M+H]+; 1H NMR (500 MHz, DMSO-de) δ ppm 3.92 (s, 2 H) 7.18 (d, J=8.54 Hz, 1 H) 7.23 - 7.31 (m, 1 H) 7.49 (s, 1 H) 12.61 (br. s., 1 H). Intermediate 43: 7-chloro-3-(methylthio)-2H-1 ,4-benzothiazine
Figure imgf000046_0002
To a solution of 7-chloro-2H-1 ,4-benzothiazine-3(4H)-thione (intermediate 42, 3 g, 12.10 mmol) in acetone (100 mL) at room temperature was added potassium hydroxide (1.697 g, 30.2 mmol) and 400 μΙ of methyl iodide. The solution was stirred for 15 minutes and then a further 400 μΙ of methyl iodide was added. The reaction mixture was stirred for 2 hours. The resulting red colored suspension was filtered, to remove the inorganic salts, and the filtrate was concentrated. The residue was purified by chromatography using a Biotage SP1 purification device using 100 g normal phase silica SNAP coloumn and a cyclohexane: EtOAc solvent system (gradient 0-10 % of EtOAc in 20 CV) to give the title compound (2 g); m/z (ES): 229.9 [M+H]+; 1 H NMR (500 MHz, CHLOROFORM-d) δ ppm 2.55 (s, 3 H) 3.23 (s, 2 H) 7.1 1 - 7.15 (m, 1 H) 7.21 (d, J=8.39 Hz, 1 H) 7.23 (d, J=2.14 Hz, 1 H).
Intermediate 44: Ethyl (2Z,2E)-(7-chloro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate
Figure imgf000047_0001
7-chloro-3-(methylthio)-2H-1 ,4-benzothiazine (Intermediate 43, 2010 mg, 8.75 mmol) was dissolved in ethyl nitroacetate (10 ml_, 88 mmol) and benzyltrimethylammonium hydroxide solution in methanol (4.17 ml_, 9.19 mmol) was added. The mixture was heated at 45 °C for 18 hours. The reaction mixture was cooled to room temperature, and was quenched with 200 ml of water. Organic substances were extracted with DCM (twice, 250 ml of DCM was used in total). DCM layers were washed with 10% K2CO3 water solution (300 ml was used in total, twice). The combined organic layers were dried over
Figure imgf000047_0002
the solvent was evaporated and the residue was purified by chromatography on Biotage SP1 purification device using 100 g normal phase silica SNAP coloumn and solvent system cyclohexane:EtOAc (gradient 0-30 % of EtOAc in 20 CV) to give the title compound (700 mg) as a mixture of two possible geometric isomers; m/z (ES-): 313.01 [M]"; 1H NMR (300 MHz, CHLOROFORM-d) Bppm 1.37 (m, 3 H) 3.76 (m, 2 H) 4.36 (m, 2 H) 6.97 (d, J=8.57 Hz, 0.5 H) 7.05 (d, J=8.52 Hz, 0.5 H) 7.14 - 7.24 (m, 1 H) 7.29 - 7.35 (m, 1 H) 1 1.48 (br. s., 0.5 H) 1 1.90 (br. s., 0.5 H)
Intermediate 45: ethyl 7-chloro-4H-[1 ,2,3ltriazolo[5,1-cl[1 ,4lbenzothiazine-3- carboxylate
Figure imgf000047_0003
To a solution of ethyl (2Z,2E)-(7-chloro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (Intermediate 44, 200 mg, 0.635 mmol) (a mixture of geometric isomers) dissolved in acetic acid (5 mL) cooled in an ice bath was added zinc (249 mg, 3.81 mmol), portionwise over 5 minutes. The temperature of the mixture was allowed to reach room temperature during next 40 minutes and then isoamyl nitrite (0.120 mL, 0.890 mmol) was added followed by trichloroacetic acid (51.9 mg, 0.318 mmol). The mixture was stirred at room temperature for 2 hours then the reaction mixture was poured into 50 ml of water. The organic substances were extracted using phase separator cartridge using 60 ml of DCM in total. The DCM layer was then washed twice with 100 ml of NaHCC>3 (in total). The DCM layer was then dried over phase separator cartridge, solvent was evaporated and the resulting crude was purified by chromatography on Biotage SP1 purification device using 10 g normal phase silica SNAP coloumn and cyclohexane:EtOAc solvent system (gradient 0-30% of EtOAc in 25 CV) to give the title compound; m/z (ES): 296.07 [M+H]+ ;1H NMR (600 MHz, CHLOROFORM-d) δ ppm 1.46 (t, J=7.15 Hz, 3 H) 4.45 (s, 2 H) 4.48 (q, J=7.15 Hz, 2 H) 7.36 (dd, J=8.64, 2.18 Hz, 1 H) 7.48 (d, J=2.27 Hz, 1 H) 8.14 (d, J=8.72 Hz, 1 H). Intermediate 46: ethyl 7-chloro-1-methyl-4H-imidazor5, 1-cin ,4lbenzothiazine-3- carboxylate
Figure imgf000048_0001
To a solution of ethyl (2Z,2E)-(7-chloro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (intermediate 44, 240 mg, 0.763 mmol) mixture of geometric isomers dissolved in acetic acid (6 mL) cooled in an ice bath was added zinc (299 mg, 4.58 mmol) portionwise over during 5 minutes. The temperature of the mixture was allowed to reach room temperature over the next 40 minutes and then triethyl orthoacetate (0.287 mL, 1 .525 mmol) was added. The mixture was stirred on room temperature for 20 hours. Then the reaction mixture was poured into 50 ml of water. Organic substances were extracted using phase separator cartridge with 60 ml of DCM in total. DCM layer was then washed twice with 100 ml of NaHCC>3 in total. DCM layer was then dried over phase separator cartridge, the solvent was evaporated and the resulting crude was purified by chromatography on Biotage SP1 purification device using 25 g normal phase silica SNAP coloumn and
cyclohexane:EtOAc solvent system (gradient 50-100% of EtOAc in 20 CV) to give the title compound (60 mg); m/z (ES): 308.90 [M+H]+ ;1H NMR (500 MHz,
CHLOROFORM-d) δ ppm 1.42 (t, J=7.17 Hz, 3 H) 2.67 (s, 3 H) 4.32 (s, 2 H) 4.42 (q, J=7.17 Hz, 2 H) 7.32 (dd, J=8.70, 2.29 Hz, 1 H) 7.46 (d, J=8.70 Hz, 1 H) 7.55 (d, J=2.29 Hz, 1 H).
Intermediate 47: ethyl 7-chloro-4H-[1 ,2,3ltriazolo[5,1-cl[1 ,4lbenzothiazine-3- carboxylate 5,5-dioxide
Figure imgf000049_0001
To a solution of ethyl 7-chloro-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3- carboxylate (intermediate 45, 58 mg, 0.196 mmol) dissolved in a mixture of dry dichloromethane (1.5 ml) and methanol (0.5 ml) under argon cooled in an ice bath was added m-CPBA (135 mg, 0.588 mmol) portionwise. The mixture was stirred at room temperature overnight (18 hours). The solvent was evaporated and the resulting residue was dissolved in 20 ml of DCM. The orgainic phase was washed with 2x30 ml of saturated sodium bicarbonate solution using a phase separator cartridge and the solvent removed. The resulting residue was purified by
chromatography using Biotage SP1 purification device, 10 g normal phase silica
SNAP coloumn and cyclohexane/EtOAc solvent system(gradient 30-60 % of EtOAc in 20 CV) to give the title compound (50 mg); m/z (ES): 328.08 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 1.48 (t, J=7.17 Hz, 3 H) 4.51 (q, J=7.12 Hz, 2 H) 5.05 (s, 2 H) 7.84 (dd, J=8.77, 2.21 Hz, 1 H) 8.07 (d, J=2A4 Hz, 1 H) 8.37 (d, J=8.70 Hz, 1 H).
Intermediate 48: 7-chloro-1-methyl-4H-imidazo[5,1 -cl[1 ,41benzothiazine-3-carboxylic acid
Figure imgf000049_0002
Ethyl 7-chloro-1-methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3-carboxylate
(intermediate 46, 36 mg, 0.1 17 mmol) was dissolved in a mixture of ethanol (4 mL) and water (1 mL, 55.5 mmol). To this mixture was added potassium hydroxide (60 mg, 0.909 mmol) and the mixture was stirred at 70 °C for 2 hours, then the mixture was evaporated to dryness. The resulting crude was dissolved in 5 ml of water, the pH of the solution was adjusted to 4.5 to 5.5 and the resulting acid was extraced with DCM using phase separator cartridge (250 ml of DCM was used in total). The solvent was evaporated to give the title compound (27 mg); m/z (ES): 281.2 [M+H]+; 1H NMR
(500 MHz, DMSO-d6) δ ppm 2.58 (s, 3 H) 4.35 (s, 2 H) 7.48 (dd, J=8.77, 2.37 Hz, 1 H) 7.75 (d, J=2.29 Hz, 1 H) 7.82 (d, J=8.70 Hz, 1 H). Intermediate 49: 5-chloro-6-fluoro-1 ,3-benzothiazol-2-amine and 7-chloro-6-fluoro- 1 ,3-benzothiazol-2-amine mixture
Figure imgf000050_0001
In a 1000 ml three-neck round bottom flask were combined 150 ml of acetic acid, 3- chloro-4-fluoroaniline (10 g, 68.7 mmol) and potassium thiocyanate (13.35 g, 137 mmol) with continous mechanical stirring. Bromine (5.31 mL, 103 mmol) in 150 ml of acetic acid, was added dropwise to the reaction mixture over 30 minutes whilst maintaining the temperature between 15 and 20 °C throughout the addition. Stirring was continued for an additional 1 h after bromine addition. The resulting suspension was filtered and dried. The precipitate was dissolved in warm water (1 litre) and basified with 25 % aqueous ammonium hydroxide solution to pH 1 1 to 12 with cooling. The resulting precipitate was filtered, washed with aqueous ammonium hydroxide to pH 1 1 to 12) and dried in a vacuum oven at 40 °C to give the title mixture of two regioisomers, 5-chloro-6-fluoro-1 ,3-benzothiazol-2-amine (6.32 g) and 7-chloro-6-fluoro-1 ,3-benzothiazol-2-amine (3.16 g);
5-chloro-6-fluoro-1 ,3-benzothiazol-2-amine: 1H NMR (500 MHz, DMSO-c/6) δ ppm 7.46 (d, J=6.71 Hz, 1 H) 7.67 (s, 2 H) 7.81 (d, J=9.16 Hz, 1 H)
5-chloro-6-fluoro-1 ,3-benzothiazol-2-amine: 1H NMR (500 MHz, DMSO-c/6) δ ppm 7.22 - 7.32 (m, 1 H) 7.74 (br. s., 1 H).
Intermediate 50: 6-chloro-7-fluoro-2H-1 ,4-benzothiazin-3(4H)-one and 8-chloro-7-
Figure imgf000050_0002
The mixture from Intermediate 49 (5 g) was dissolved in 50 % NaOH water solution (70 mL) and heated at 130 °C for 16 hours. The mixture was poured on to 300 ml of ice. The pH was adjusted to 7 to 7.5 with glacial acetic acid whilst stirring vigorously with cooling in an ice/acetone bath. The mixture was extracted with diethyl ether (1 litre) and the combined diethyl ether layers were dried over IS^SC^. The residue was dissolved in 50 ml of THF under an Argon atmosphere (solution A). In another, ice cooled, round bottom three-neck flask under argon atmosphere was prepared a suspension of lithium bromide (1.737 g, 20 mmol) and sodium borohydride (0.757 g, 20 mmol) in 50 ml of dry THF. The suspension was stirred at 0 °C for 15 minutes. Solution A was added dropwise to the suspension. The temperature of the mixture was raised to 35 °C and was stirred for 1 hour. To the mixture was added sodium ethoxide (1 .528 g, 22 mmol) followed by methyl bromoacetate (2.090 mL, 22 mmol). The mixture was stirred overnight at room temperature and 200 ml of water was added. The mixture was extracted with DCM (200 ml was used in total) and the DCM layer was dried using a phase separator cartridge. The solvent was evaporated and the residue was dissolved in dimethyl sulfoxide (200 mL) and heated at 70 °C for 16 hours. The mixture was cooled to room temperature, poured in 200 ml of water and the resulting precipitate was filtered and dried in a vacuum oven at 40 °C. Purification by chromatography on Biotage SP1 purification system using 340 g normal phase silica SNAP coloumn using cyclohexane/EtOAc solvent system (gradient 20-55 % in 20 CV) gave 6-chloro-7-fluoro-2H-1 ,4-benzothiazin-3(4H)-one (1 .7 g); m/z (ES): 218.1 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 3.51 (s, 2 H) 7.09 (d, J=6.87 Hz, 1 H) 7.52 (d, J=9.31 Hz, 1 H) 10.66 (s, 1 H); and 8-chloro-7-fluoro-2H-1 ,4- benzothiazin-3(4H)-one (1 g); m/z (ES): 218.1 [M+H]+; 1H NMR (500 MHz, DMSO-d6) □ ppm 3.57 (s, 2 H) 6.92 - 6.98 (m, 1 H) 7.22 - 7.30 (m, 1 H) 10.76 (s, 1 H).
Intermediate 51 : 6-chloro-7-fluoro- -1 ,4-benzothiazine-3(4H)-thione
Figure imgf000051_0001
In a 25 ml microwave crucible were suspended mixture of 6-chloro-7-fluoro-2H-1 ,4- benzothiazin-3(4H)-one (1.6 g, 7.35 mmol) and Lawesson's reagent (1.7 g, 4.20 mmol) in dry toluene (15 mL) and the suspension was heated at 120 °C for 7 minutes at 450 Watts (following prestirring for 1 minute). The resulting precipitate was filtered and rinsed with DCM (70 ml in total, portionwise). The resulting material was dried in a vacuum oven at 40 °C to give the title compound (1.44 g); m/z (ES): 234.1 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 3.95 (s, 1 H) 7.33 (d, J=6.87 Hz, 1 H) 7.59 (d, J=9.31 Hz, 1 H) 12.60 (s, 1 H). Intermediate 52: 6-chloro-7-fluoro-3-(methylthio)-2H-1 ,4-benzothiazine
Figure imgf000051_0002
To a solution of 6-chloro-7-fluoro-2H-1 ,4-benzothiazine-3(4H)-thione (intermediate 51 , 1.44 g, 6.16 mmol) in acetone (70 ml) was added potassium hydroxide (0.864 g, 15.40 mmol) and methyl iodide (0.212 ml, 3.39 mmol). The mixture was stirred for 15 minutes and then further methyl iodide (0.212 ml, 3.39 mmol) was added. The reaction mixture was stirred for 1 hour and the solvent was removed. To the residue was added 100 ml of DCM and 100 ml of water. Using phase separator cartridge, the DCM layer was collected, and the solvent was evaporated. The residue was purified on Biotage SP1 purification device using cyclohexane/EtOAc solvent system (gradient 0-8 % in 20 CV) to give the title compound (1.25 g); m/z (ES): 248.1
[M+H]+; 1H NMR (600 MHz, CHLOROFORM-d) δ ppm 2.54 (s, 3 H) 3.24 (s, 2 H) 7.04 (d, J=8.72 Hz, 1 H) 7.36 (d, J=7.15 Hz, 1 H).
Intermediate 53: ethyl (2Z,2E)-(6-chloro-7-fluoro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate
Figure imgf000052_0001
6-Chloro-7-fluoro-3-(methylthio)-2H-1 ,4-benzothiazine (intermediate 52, 1.25 g, 5.05 mmol) was dissolved in ethyl nitroacetate (6 ml, 54.2 mmol) under an argon atmosphere. Benzyltrimethylammonium hydroxide in 40 % methanol solution (2.316 ml, 5.10 mmol) was added and the resulting mixture was stirred for 48 hours at 40 °C. The mixture was diluted with water (approximately 100 ml). Organic substances were extracted with 3x50 ml of DCM and the combined DCM layers were washed with 2x50 ml of 10 % aqueous potassium carbonate solution. The DCM layers were dried using a phase separator, the solvent was evaporated and the residue was purified by chromatography using a Biotage SP1 purification device, using cyclohexane/EtOAc solvent system and 50 g normal phase silica SNAP coloumn (gradient 1-15% of EtOAc in 20 CV) to give the title compound (267 mg); m/z (ES): 333.3 [M+H]+; 1H NMR (300 MHz, CHLOROFORM-d) δ ppm 1.34-1 .39 (m, 3 H) 3.76- 3.78 (m, 2 H) 4.33-4.40 (m, 2 H) 7.1 1 - 7.21 (m, 2 H) 1 1.46 (br. s., 0.5 H) 1 1 .84 (br. s., 0.5 H).
Intermediate 54: Ethyl 8-chloro-7-fluoro-4H-ri,2,3ltriazolor5,1-cin,4lbenzothiazine-3- carboxylate
Figure imgf000053_0001
To a vigorously stirred solution of ethyl (2Z,2E)-(6-chloro-7-fluoro-2H-1 ,4- benzothiazin-3(4H)-ylidene)(nitro)ethanoate (intermediate 53, 80 mg, 0.240 mmol) in acetic acid (5 ml) cooled in ice to between 0 and 5 °C, under an argon atmosphere was added zinc (94 mg, 1.443 mmol) over a 5 minute period. The temperature of the mixture was allowed to reach room temperature and the mixture was stirred for 30 minutes. Isoamyl nitrite (0.045 ml, 0.337 mmol) and trichloroacetic acid (19.64 mg, 0.120 mmol) were added and the resulting suspension was stirred for 1 hour at room temperature. The reaction was quenched with 10 ml of water and the mixture was extracted with 100 ml of DCM using a phase separator cartridge. The DCM solution was washed with 100 ml of saturated aqueous sodium bicarbonate solution, and evaporated. The residue was purified by chromatography on a 5 g normal phase silica ISOLUTE Si column, eluting with 50 ml of cyclohexane:EtOAc (5:1 ) to give the title compound (78 mg); m/z (ES): 314.2 [M+H]+; 1H NMR (500 MHz,
CHLOROFORM-d) δ ppm 1.46 (t, J=7.17 Hz, 3 H) 4.44 - 4.53 (m, 4 H) 7.29 (d, J=8.39 Hz, 1 H) 8.30 (d, J=6.71 Hz, 1 H).
Intermediate 55: Ethyl 8-chloro-7-fluoro-1-methyl-4H-imidazor5,1 - cl Γ 1 ,41 benzoth iazi n e-3-ca rboxylate
Figure imgf000053_0002
To a stirred solution of ethyl (2Z/2E)-(6-chloro-7-fluoro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (intermediate 53, 100 mg, 0.301 mmol) in glacial acetic acid (5 mL) cooled in ice under an argon atmosphere was added zinc (1 18 mg, 1 .803 mmol) portionwise over 5 minutes. The mixture was warmed to room temperature and stirred for 1 h. The mixture was filtered and the filtrated was evaporated under reduced pressure to give a residue. The residue was dissolved in 50 ml of DCM and treated with 100 ml of saturated aqueous sodium bicarbonate solution. The organic phase was separated using a phase separator cartridge and the solvent was evaporated. The residue was suspended in triethyl orthoacetate (1 ml, 5.42 mmol) and p-toluenesulfonic acid monohydrate (10 mg, 0.053 mmol) was added and the mixture was heated at 90 °C for 4 hours and stirred on room temperature for 60 hours. The product was puriifed by chromatography using a Biotage SP1 purification device, 25 g normal phase silica SNAP coloumn and a cyclohexane/EtOAc solvent system (gradient 3-20% of EtOAc in 3CV + 20-40% of EtOAc in 9 CV + 50-80 % of EtOAc in 12 CV) to give the title compound (37 mg); m/z (ES): 327.2 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 1.42 (t, J=7.10 Hz, 3 H) 2.69 (s, 3 H) 4.33 (s, 2 H) 4.42 (q, J=7.17 Hz, 2 H) 7.36 (d, J=8.39 Hz, 1 H) 7.57 (d,J=6.41 Hz, 1 H). Intermediate 56: Ethyl 8-chloro-7-fluoro-1-methyl-4H-imidazor5,1 - clH ,41benzothiazine-3-carboxylate 5,5-dioxide
Figure imgf000054_0001
To a solution of ethyl 8-chloro-7-fluoro-1-methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine- 3-carboxylate (intermediate 55, 34 mg, 0.095 mmol) in dichloromethane (dried using a phase separator) (3 mL) and absolute ethanol (300 μΙ) ice cooled under an argon atmosphere was added m-cpba (55 mg, 0.223 mmol) portionwise. The mixture was stirred at room temperature for 18 hours. The mixture was washed with saturated aqueous sodium bicarbonate solution (3x25 ml) and using a phase separator cartridge, the mixture was dried. The solvent was evaporated and the residue was triturated with diethyl ether to give the title compound (33 mg); m/z (ES): 359.3
[M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 1.44 (t, J=7.10 Hz, 3 H) 2.79 (s, 3 H) 4.45 (q, J=7.07 Hz, 2 H) 4.99 (s, 2 H) 7.70 (d, J=5.49 Hz, 1 H) 7.91 (d, J=7.02 Hz, 1 H).
Figure imgf000054_0002
In a 2L three-necked round bottom flask was dissolved 4-fluoroaniline (17.29 mL, 180 mmol) in acetic acid (300 mL) under vigourous mechanical stirring at room temperature. Potassium thiocyanate (35.0 g, 360 mmol) was added in portions, and stirring was continued for 10 minutes. Bromine (13.91 mL, 270 mmol) in acetic acid (300 mL) was added dropwise to the reaction mixture keeping the temperature under 30 °C (cooling in ice bath). After bromine addition, stirring was continued for an additional 1 h and the resulting precipitate was collected by filtration. The filter cake was washed several times with water (3 L of water was used in total), and the aqueous solution was filtered. The filtrate was basified with 25% aqueous ammonium hydroxide to pH 10 to1 1 . The resulting precipitate was collected and dried under vaccum at 40 °C overnight to give the title compound (24.5 g); 1H NMR (500 MHz, DMSO-de): δ ppm 7.03 (td, J=9.08, 2.59 Hz, 1 H) 7.29 (dd, J=8.70, 4.88 Hz, 1 H) 7.45 (s, 2 H) 7.57 (dd, J=8.77, 2.67 Hz, 1 H); MS (m/z): 169 [MH]+
Intermediate 58: 2-Amino-5-fluorobenzenethiol
6-Fluoro-1 ,3-benzothiazol-2-amine (intermediate 57, 12 g, 71.3 mmol) was suspended in 50 % aquous sodium hydroxide soluiton (200 mL) and heated under reflux for 24 hours. The mixture was cooled, diluted with water (70 mL) and extracted with ethyl acetate (3x). The organic phase was washed with water and all aqueous layers (3 L in total) were combined, filtered and the filtrate was acidified with glacial acetic acid to pH 6). This aqueous mixture was extracted with ethyl-acetate (3x) and the combined extracts were washed with water and dried over IS^SC^. The solvent was evaporated to give the title compound (6.2 g); 1H NMR (500 MHz,
CHLOROFORM-d): δ ppm 3.06 (br. s., 1 H) 3.95 (br. s., 2 H) 6.67 (dd, J=8.70, 4.88 Hz, 1 H) 6.82 (td, J=8.43, 2.82 Hz, 1 H) 7.09 (dd, J=8.47, 2.82 Hz, 1 H); MS (m/z): 144 [MH]+.
Figure imgf000055_0001
To a solution of 2-amino-5-fluorobenzenethiol (intermediate 58, 3 g, 20.95 mmol) in dimethyl sulfoxide (70 mL) at room temperature was added sodium ethoxide (1.568 g, 23.04 mmol). After half hour, methyl bromoacetate (2.124 mL, 23.05 mmol) was added and the mixture was stirred overnight at room temperature. The solution was poured into ice water and acidified with 3 M HCI solution to pH 5. The resulting precipitate was collected by filtration, washed with water and dried to give the title compound (3.2 g); 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 3.43 (s, 2 H) 6.83 - 6.87 (m, 1 H) 6.87 - 6.92 (m, 1 H) 7.05 (dd, J=8.16, 2.67 Hz, 1 H) 8.92 (br. s., 1 H); MS (m/z): 184 [MH]+.
Figure imgf000056_0001
Two parallel reactions were performed in a microwave reactor. In the first reaction, the reaction vessel containing 7-fluoro-2H-1 ,4-benzothiazin-3(4H)-one (intermediate 59, 1 ,5 g, 8,19 mmol) and Lawesson's reagent (1 ,987 g, 4,91 mmol) suspended in toluene (16 mL) was sealed and heated in a Biotage Initiator at 150 °C for 7 minutes. The second reaction vessel also containing 7-fluoro-2H-1 ,4-benzothiazin-3(4H)-one (1 ,7 g, 9,28 mmol) and Lawesson's reagent (2,252 g, 5,57 mmol) suspended in toluene (18 mL) was sealed and heated in Biotage Initiator at 150 °C for 7 minutes. After cooling, the two reaction mixtures were gathered and triturated with
cyclohexane. The resulting solids were collected and washed with DCM to give the title compound (1 .8 g); 1H NMR (500 MHz, DMSO-d6): δ ppm 3.91 (s, 2 H) 7.08 (td, J=8.66, 2.67 Hz, 1 H) 7.21 (dd, J=8.85, 5.19 Hz, 1 H) 7.32 (dd, J=8.77, 2.67 Hz, 1 H) 12.56 (br. s., 1 H); MS (m/z): 200 [MH]+; The mother liquor from the trituration was evaporated and the residue was purified using a Biotage SP-1 system using 50 g Si SNAP column eluting with cyclohexane / ethyl-acetate gradient (10-20 % of ethyl- acetate / 20 CV) to give a second batch of the title compound (0.4 g); 1H NMR (500 MHz, DMSO-d6): δ ppm 3.91 (s, 2 H) 7.09 (td, J=8.62, 2.75 Hz, 1 H) 7.21 (dd, J=8.85, 5.19 Hz, 1 H) 7.32 (dd, J=8.85, 2.75 Hz, 1 H) 12.56 (br. s., 1 H); MS (m/z): 200 [MH]+ Intermediate 61 : 7-Fluoro-3-(methylthio)-2H-1 ,4-benzothiazine
Figure imgf000056_0002
To a solution of 7-fluoro-2H-1 ,4-benzothiazine-3(4H)-thione (intermediate 60
2.17 g, 10.89 mmol) in acetone (65 mL) at room temperature was added potassium hydroxide (1.527 g, 27.2 mmol) followed by methyl iodide (0.749 mL, 1 1.98 mmol) which was added portionwise over 15 minutes. The reaction mixture was stirred at room temperature for 2 hours. Solvent was evaporated and the resulting residue was extracted with DCM, water and brine. The combined organic layers were dried over a phase separator filter tube and concentrated under vacuum. The residue was purified on a Biotage SP-1 system using 100 g Si SNAP column, eluting with cyclohexane / ethyl-acetate gradient (0-20 % of ethyl-acetate / 20 CV) to give the title compound (1 .97 g); 1H NMR (500 MHz, DMSO-d6): δ ppm 2.49 (s, 3 H) 3.49 (s, 2 H) 7.04 (td, J=8.62, 2.90 Hz, 1 H) 7.21 - 7.29 (m, 2 H); 1H NMR (500 MHz,
CHLOROFORM-d): δ ppm 2.55 (s, 3 H) 3.24 (s, 2 H) 6.88 (td, J=8.47, 2.75 Hz, 1 H) 6.96 (dd, J=8.47, 2.82 Hz, 1 H) 7.22 - 7.29 (m, 1 H, overlaped with CDCI3; MS (m/z): 214 [MH]+
Intermediate 62: Ethyl (2Z/2E)-(7-fluoro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (equal mixture of geometric isomers)
Figure imgf000057_0001
To a solution of 7-fluoro-3-(methylthio)-2H-1 ,4-benzothiazine (intermediate 61 , 1.96 g, 9.19 mmol) in ethyl nitroacetate (10.20 ml, 92 mmol) at room temperature and under argon stream, Triton-B 40 % in MeOH (4.18 ml, 9.19 mmol) was added and the mixture heated to 50 °C and stirred at that temperature for 24 hours. Further (0.2 eq) of Triton-B 40 % in MeOH (0.84 ml, 1.84 mmol) was added and the reaction mixture was stirred at 50 °C overnight. The reaction mixture was cooled to room temperature, quenched with water and extracted with DCM (2x). The combined organic layers were washed with 10 % aqueous potassium carbonate solution (2x), then dried over a phase separator filter tube and concentrated under reduced pressure. The obtained crude was purified on a Biotage SP-1 system using 100 g Si SNAP column eluting with cyclohexane / ethyl-acetate gradient (0-30 % of ethyl- acetate / 20 CV) to give the title compound (1.15 g) (obtained as an equal mixture of geometric isomers; as determined by 1H NMR spectrum); 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 1.33 - 1.40 (m, 3 H) 3.77 (m, 2 H) 4.31 - 4.41 (m, 2 H) 6.90 - 7.13 (m, 3 H) 1 1.53 (br. s., 0.5 H) 1 1.98 (br. s., 0.5 H); MS (m/z): 299 [MH]+ Intermediate 63: Ethyl 7-fluoro-4H-ri.2,31triazolor5,1-ciri.41benzothiazine-3- carboxylate
Figure imgf000058_0001
To a solution of ethyl (2Z/2E)-(7-fluoro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (intermediate 62, 400 mg, 1.341 mmol) (equal mixture of geometric isomers) dissolved in acetic acid (10 ml_), cooled in ice was added zinc (526 mg, 8.05 mmol) portionwise over 5 minutes. The mixture was warmed to room temperature and stirred for 45 minutes. Isoamyl nitrite (0.253 ml_, 1.877 mmol) was added followed by trichloroacetic acid (1 10 mg, 0.670 mmol) and the mixture was stirred at room temperature for 2.5 hours. The reaction was quenched with water and extracted with DCM (2x). The combined organic phase was washed twice with a saturated solution of sodium bicarbonate, dried over a phase separator filter tube and concentrated under reduced pressure. The resulting residue was purified using a Biotage SP-1 system on a 25 g Si SNAP column eluting with a cyclohexane / ethyl- acetate gradient (0-30 % of ethyl-acetate / 20 CV) to give the title compound (270 mg); 1H NMR (600 MHz, CHLOROFORM-d): δ ppm 1.46 (t, J=7.15 Hz, 3 H) 4.46 (s, 2 H) 4.48 (q, J=7.15 Hz, 2 H) 7.09 (ddd, J=8.77, 7.89, 2.88 Hz, 1 H) 7.20 (dd, J=8.20, 2.62 Hz, 1 H) 8.19 (dd, J=8.90, 5.06 Hz, 1 H); MS (m/z): 280 [MH]+ Intermediate 64: Ethyl 7-fluoro-4H-ri.2,31triazolor5,1-ciri.41benzothiazine-3- carboxylate 5,5-dioxide
Figure imgf000058_0002
To a solution of ethyl 7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3- carboxylate (intermediate 63, 90 mg, 0.322 mmol) in a mixture of dry
dichloromethane (DCM) (4 mL) and dry methanol (1 mL) was added mCPBA (167 mg, 0.967 mmol) and the reaction mixture was stirred at room temperature overnight. Further (0.5 eq) mCPBA (28 mg, 0.161 mmol) was added and the mixture was stirred at room temperature for 24 hours. Evaporation of the solvent gave a residue which was dissolved in DCM and washed with saturated sodium bicarbonate solution. The organic layer was dried using a phase separator filter tube and concentrated under reduced pressure. The residue was purified using a Biotage SP-1 system using 25 g Si SNAP column eluting with cyclohexane / ethyl-acetate gradient (0-40 % of ethyl- acetate / 20 CV; 40 % of ethyl-acetate / 5 CV) to give the title compound (80 mg); 1H NMR (500 MHz, DMSO-d6): δ ppm 1 .37 (t, J=7.10 Hz, 3 H) 4.41 (q, J=7.02 Hz, 2 H) 5.56 (s, 2 H) 7.92 (td, J=8.70, 2.75 Hz, 1 H) 8.08 (dd, J=7.32, 2.75 Hz, 1 H) 8.49 (dd, J=9.00, 4.27 Hz, 1 H); MS (m/z): 312 [MH]+
Intermediate 65: Ethyl 7-fluoro-1-methyl-4H-imidazor5, 1-ciri ,4lbenzothiazine-3- carboxylate
Figure imgf000059_0001
To a solution of ethyl (2Z/2E)-(7-fluoro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (Intermeidate 62, 400 mg, 1.341 mmol) (equal mixture of geometric isomers) dissolved in acetic acid (1 1 mL), cooled in ice was added zinc (526 mg, 8.05 mmol) portionwise over 10 minutes. The mixture was allowed to reach room temperature and stirred for 1 hour. Triethyl orthoacetate (0.494 mL, 2.68 mmol) was added and the mixture was stirred at room temperature overnight. The reaction mixture was quenched with water and extracted with DCM (2x). The organic phase was washed twice with saturated sodium bicarbonate solution, dried using a phase separator filter tube and concentrated under reduced pressure. The residue was purified using a Biotage SP-1 system using 50 g Si SNAP column, eluting with cyclohexane / ethyl-acetate gradient (30-70 % of ethyl-acetate / 20 CV) to give the title compound (95 mg); 1H NMR (500 MHz, DMSO-d6): δ ppm 1.30 (t, J=7.10 Hz, 3 H) 2.58 (s, 3 H) 4.26 (q, J=7.07 Hz, 2 H) 4.36 (s, 2 H) 7.29 (td, J=8.62, 2.90 Hz, 1 H) 7.58 (dd, J=8.70, 2.90 Hz, 1 H) 7.86 (dd, J=9.00, 5.04 Hz, 1 H); MS (m/z): 293 [MH]+
Intermediate 66: 7-Fluoro-1-methyl-4H-imidazo[5,1-cl[1 ,41benzothiazine-3-carboxylic acid
To a solution of ethyl 7-fluoro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3- carboxylate (intermediate 65, 80 mg, 0.274 mmol) in a mixture of tetrahydrofuran (THF) (2.5 mL) and water (2.5 mL) was added potassium hydroxide (19.96 mg, 0.356 mmol) at room temperature and the mixture was stirred first at room temperature for 3 hours then at 50 °C overnight. The mixture was allowed to cool to room
temperature and the THF evaporated. The remaining aqueous solution was adjusted to pH 5 with 1 N HCI and the mixture was extracted with DCM. The organic extracts were dried using a phase separator filter tube and concentrated under reduced pressure to give the title compound (57 mg); 1H NMR (500 MHz, DMSO-c/6): δ ppm 2.57 (s, 3 H) 4.35 (s, 2 H) 7.29 (td, J=8.58, 2.67 Hz, 1 H) 7.57 (dd, J=8.62, 2.52 Hz, 1 H) 7.85 (dd, J=8.93, 4.96 Hz, 1 H) 12.52 (br. s., 1 H); MS (m/z): 265 [MH]+
Intermediate 67: (7-Fluoro-1 -methyl-4H-imidazo[5,1 -cl[1 ,4lbenzothiazin-3-yl)carbonyl
2-methylpropyl carbonate
Figure imgf000060_0001
To a solution of 7-fluoro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylic acid (intermediate 66, 29 mg, 0.1 10 mmol) in dry dichloromethane (DCM) (5 mL) were added DIPEA (0.021 mL, 0.121 mmol) and isobutyl chloroformate (0.016 mL, 0.121 mmol) at -15 °C and stirred at -15 °C for 15 minutes and then at room temperature for 5 hours. The mixture was washed with saturated sodium bibarbonate solution, dried using a phase separator filter tube and concentrated under reduced pressure to give the title compound (39 mg); 1H NMR (300 MHz, CHLOROFORM-c/): δ ppm 1.00 (d, J=6.69 Hz, 6 H) 2.07 (dt, J=13.47, 6.73 Hz, 1 H) 2.68 (s, 3 H) 4.1 1 (d, J=6.69 Hz, 2 H) 4.29 (s, 2 H) 7.08 (ddd, J=8.95, 7.72, 2.83 Hz, 1 H) 7.30 (dd, 1 H) 7.52 (dd, J=8.95, 4.80 Hz, 1 H); MS (m/z): 365 [MH]+
Intermediate 68: 8-Chloro-7-fluoro-2H-1 ,4-benzothiazine-3(4H)-thione
Figure imgf000060_0002
8-Chloro-7-fluoro-2H-1 ,4-benzothiazin-3(4H)-one (Intermediate ?) (1 .00 g, 4.59 mmol) and Lawesson's reagent (1.1 15 g, 2.76 mmol) were suspended in toluene (12 mL) sealed in a microware vessel and heated in Biotage Initiator at 120 °C for 7 minutes. The mixture was triturated with DCM (150 mL), and the resulting precipitate was collected and dried to give the title compound (0.85 g); 1H NMR (500 MHz,
DMSO-de): δ ppm 4.01 (s, 2 H) 7.17 - 7.22 (m, 1 H) 7.28 - 7.34 (m, 1 H) 12.70 (br. s., 1 H); MS (m/z): 234 [MH]+. The mother liquor from trituation was evaporated and the residue purified using a Biotage SP-1 system using 50 g Si SNAP column, eluting with cyclohexane / ethyl-acetate gradient (10-20 % of ethyl-acetate / 20 CV) to give a second crop of the title compound (0.22 g); 1H NMR (500 MHz, DMSO-c/6): δ ppm 4.01 (s, 2 H) 7.17 - 7.22 (m, 1 H) 7.28 - 7.34 (m, 1 H) 12.70 (br. s., 1 H); MS (m/z): 234 [MH]+
Intermediate 69: 8-Chloro-7-fluo -3-(methylthio)-2H-1 ,4-benzothiazine
Figure imgf000061_0001
To a solution of 8-chloro-7-fluoro-2H-1 ,4-benzothiazine-3(4H)-thione (intermediate 68, 1.07 g, 4.58 mmol) in acetone (40 ml) at room temperature was added potassium hydroxide (0.642 g, 1 1.45 mmol) followed by methyl iodide (0.315 ml, 5.04 mmol) which was added portionwise over 15 minutes. The reaction mixture was stirred at room temperature for 2 h and filtered. The filtrate was evaporated and the residue was purified using a Biotage SP-1 system using 50 g Si SNAP column, eluting with a cyclohexane / ethyl-acetate gradient (0-20 % of ethyl-acetate / 20 CV) to give the title compound (1 .07 g); 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 2.55 (s, 3 H) 3.28 (s, 2 H) 6.98 (t, J=8.70 Hz, 1 H) 7.20 (dd, J=8.85, 5.04 Hz, 1 H); MS (m/z): 248 [MH]+
Intermediate 70: Ethyl (2Z/2E)-(8-chloro-7-fluoro-2H-1 ,4-benzothiazin-3(4H)- ylidene)(nitro)ethanoate (mixture of geometric isomers)
Figure imgf000062_0001
To a suspension of 8-chloro-7-fluoro-3-(methylthio)-2H-1 ,4-benzothiazine
(Intermediate 69, 1 .07 g, 4.32 mmol) in ethyl nitroacetate (4.79 ml, 43.2 mmol) at room temperature under argon was added Triton-B 40 % in MeOH (2.159 ml, 4.75 mmol) and the mixture stirred at 50 °C for 44 hours. On cooling, water was added and the mixture was extracted with DCM (2x). The organic layers were washed with 10 % aqueous potassium carbonate (2x), then dried using a phase separator filter tube and concentrated under reduced pressure. The residue was purified using a Biotage SP-1 system using 50 g Si SNAP column eluting with cyclohexane / ethyl- acetate gradient (0-30 % of ethyl-acetate / 25 CV) to give the title compound (240 mg) as mixture of geometric isomers in ratio 55:45 ratio as determined by proton NMR; 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 1.33 - 1.41 (m, 3 H) 3.80 (s,
I .10 H) 3.82 (s, 0.90 H) 4.35 (q, 1.10 H) 4.39 (q, 0.9 H) 6.93 - 7.09 (m, 2 H) 1 1 .52 (br. s., 0.55 H) 1 1 .91 (br. s., 0.45 H); 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 1.33 - 1.41 (m, 3 H) 3.79 - 3.83 (m, 2 H) 4.32 - 4.42 (m, 2 H) 6.93 - 7.09 (m, 2 H)
I I .48 - 1 1.95 (m., 1 H); MS (m/z): 333 [MH]+
Intermediate 71 : Ethyl 6-chloro-7-fluoro-4H-[1 ,2,3ltriazolo[5,1 -cl[1 ,4lbenzothiazine-3- carboxylate
Figure imgf000062_0002
To a vigorously stirred ice cooled solution of ethyl (2Z/2E)-(8-chloro-7-fluoro-2H-1 ,4- benzothiazin-3(4H)-ylidene)(nitro)ethanoate (intermediate 70, 100 mg, 0.301 mmol) dissolved in acetic acid (5 mL) under argon was added zinc (1 18 mg, 1 .803 mmol) portionwise over 10 minutes and the reaction mixture was stirred at room
temperature for 1 hour. Isoamyl nitrite (0.057 mL, 0.421 mmol) was added followed by trichloroacetic acid (24.55 mg, 0.150 mmol) and the mixture was stirred at room temperature for 1 .5 hours. The reaction mixture was quenched with water and extracted with DCM (2x). The organic phase was washed twice with saturated aqueous sodium bicarbonate solution, dried using a phase separator filter tube and concentrated under reduced pressure. The residue was purified using a Biotage SP- 1 system using a 10 g Si SNAP column eluting with cyclohexane / ethyl-acetate gradient (0-30 % of ethyl-acetate / 20 CV) to give the title compound (70 mg); 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 1 .46 (t, J=7.10 Hz, 3 H) 4.46 - 4.52 (m, 4 H) 7.17 - 7.22 (m, 1 H) 8.15 (dd, J=9.08, 4.65 Hz, 1 H); MS (m/z): 314 [MH]+
Intermediate 72: Ethyl 6-chloro-7-fluoro-1-methyl-4H-imidazor5,1 - cl Γ 1 ,41 benzoth iazi n e-3-ca rbox late
Figure imgf000063_0001
To a vigorously stirred ice cooled solution of ethyl (2Z/2E)-(8-chloro-7-fluoro-2H-1 ,4- benzothiazin-3(4H)-ylidene)(nitro)ethanoate (intermediate 70, 130 mg, 0.391 mmol) (mixture of geometric isomers) in glacial acetic acid (6 mL) under argon was added zinc (153 mg, 2.344 mmol) portionwise over 10 minutes and the reaction mixture was stirred at room temperature for 1 hour. The mixture was filtered and the the acetic acid was removed under reduced pressure. Triethyl orthoacetate (1 .3 mL, 7.05 mmol) was added followed by p-toluenesulfonic acid monohydrate (14.0 mg, 0.074 mmol) and the mixture was heated at 70 °C with stirring for 2 hours. The reaction was quenched with water and extracted with DCM (2x). Organic phase was washed with saturated aqueous sodium bicarbonate solution, dried using a phase separator filter tube and concentrated under reduced pressure. The resulting residue was purified using a Biotage SP-1 system using 25 g Si SNAP column, eluting with cyclohexane / ethyl-acetate gradient (30-100 % of ethyl-acetate / 25 CV) to give the title compound (52 mg); 1H NMR (600 MHz, CHLOROFORM-d): δ ppm 1.43 (t, J=7.15 Hz, 3 H) 2.66 (s, 3 H) 4.35 (s, 2 H) 4.43 (q, J=7.04 Hz, 2 H) 7.16 (t, J=8.46 Hz, 1 H) 7.43 (dd, J=8.98, 4.45 Hz, 1 H); MS (m/z): 327 [MH]+
Compound 1 : 8-chloro-N-(2-methyl-4-pyridinylV4H-ri ,2.31ΐήβζοΙοΓ5.1 - ciri,41benzothiazine-3-carboxamide
Figure imgf000063_0002
To a solution of 2-methyl-4-pyridinamine (73.1 mg, 0.676 mmol) in 1 ,4-dioxane (2 mL) stirred under argon at room temperature was added a solution of
trimethylaluminium (0.338 mL, 0.676 mmol) in heptane dropwise during 5 min and the reaction mixture was stirred at room temperature for 1 hr. A solution of ethyl 8- chloro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxylate (Intermediate 5) (50 mg, 0.169 mmol) in 1 ,4-dioxane (2 mL) was added dropwise at room temperature under argon. The resulting mixture was heated to 100 °C and stirred at that temperature for 15 hours. The reaction was quenched with dioxane/water (30/1 )and the reaction mixture was evaporated to dryness. DCM (50 ml) was added and stirred for 0.5 h. NaHCC>3 (sat. sol.) (20 ml) was added and stirring was continued for 15 min. The layers were separated by passing through an 1ST Phase separator, and the aqueous layer was extracted three times with DCM (3x50ml). The combined organic layers were then washed with water (100 ml) and brine (100ml), dried over sodium sulphate and passed through an 1ST phase separator. The solvent was removed and the residue was purified on Biotage SP1 , Snap Si 25 g, 25 ml/min using a gradient of 2N NH3/MeOH in DCM: 1 % of 2N NH3/MeOH to give the desired compound (47.5 mg); 1H NMR (500 MHz, CHLOROFORM-d) d ppm 2.59 (s, 3 H) 4.55 (s, 2 H) 7.33 - 7.38 (m, 1 H) 7.44 (d,J=8.54 Hz, 2 H) 7.54 (d, J=1.53 Hz, 1 H) 8.21 (d, J=2.14 Hz, 1 H) 8.47 (d, J=5.49 Hz, 1 H) 9.00 (s, 1 H); m/z: 358 [M+1 ].
Compound 2: 8-chloro-N-(2-chloro-4-pyridinylV4H-ri .2,3ltriazolor5,1- ciri,41benzothiazine-3-carboxamide
Figure imgf000064_0001
To a solution of 2-chloro-4-pyridinamine (21.73 mg, 0.169 mmol) in 1 ,4-dioxane (2 mL) stirred under argon at room temperature was added a solution of
trimethylaluminium (0.338 mL, 0.676 mmol) in heptane dropwise during 5 min. The reaction mixture was stirred at room temperature for 1 hr. A solution of ethyl 8-chloro- 4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxylate (intermediate 5) (50 mg, 0.169 mmol) in 1 ,4-dioxane (2 mL) was added dropwise at room temperature under argon. The resulting mixture was heated to 90 °C and stirred at that temperature for 15 hours. The reaction was then cooled to room temperature and then to 0°C and quenched with dioxane/water (30/1 ). The entire reaction mixture was evaporated to dryness. DCM (50 ml) was added and stirred for 0.5 h. NaHC03 (sat. sol.) (20 ml) was added and stirring continued for 15 min. The layers were separated by passing the mixture through an 1ST Phase separator. The aqueous layer was extracted three times with DCM (3x50ml) and the combined organic layers were washed with water (100 ml) and brine (100ml), dried over sodium sulphate and passed through an 1ST phase separator. The residue was suspended in DC M/MeO 1-1:98/2 (25 ml) and the resulting precipitate was filtered and after dried to give the title compound (10 mg). The filtrate was put on a Snap Si samplet 10g and purified on Biotage SP1 Snap Si 50 g; 40 ml/min by eluting with 40% of EtOAc in cyclohexane to give more of the title compound (21 mg); 1H NMR (500 MHz, DMSO-d6) d ppm 4.55 - 4.81 (m, 1 H) 7.51 - 7.61 (m, 1 H) 7.65 - 7.75 (m, 1 H) 7.82 - 7.95 (m, 1 H) 8.01 - 8.10 (m, 1 H) 8.13 - 8.22 (m, 1 H) 8.28 - 8.39 (m, 1 H) 1 1.21 - 1 1.50 (m, 1 H); m/z: 358 [M+1].
Compound 3: 8-chloro-1 -methyl-/V-(2-methyl-4-pyridinyl)-4H-imidazo[5, 1 - clH ,41benzothiazine-3-carboxamide 5,5-dioxide
Figure imgf000065_0001
To a suspension of 8-chloro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3- carboxylic acid 5,5-dioxide (obtained by mCPBA oxidation of Intermediate 7) (30 mg, 0.096 mmol) in dichloromethane (1 mL) stirred at -10 °C was added TEA (0.015 mL, 0.106 mmol) under an argon atmosphere. The reaction mixture was stirred at -10 °C until a clear solution formed. Isobutyl chloroformate (0.014 mL, 0.106 mmol) was added dropwise over 5 min and the reaction mixture was allowed to warm to room temperature and stirred overnight (Reaction mixture 1 ). To a solution of 2-methyl-4- pyridinamine (41 .5 mg, 0.384 mmol) in 1 ,4-dioxane (1 mL) stirred at room
temperature was added dropwise trimethylaluminium (0.192 mL, 0.384 mmol) over 1 min and mixture was stirred for 1 hour at room temperature. This pale yellow solution was cannulated dropwise on to Reaction mixture 1 and the combined mixture was stirred for 16 hours at rt. Water (15 ml) was added, followed by 1 N NaOH (10ml) and DCM (50 ml). The resulting suspension was stirred for 30min. The layers were separated and the aqueous layre was washed with DCM (2x30ml). The combined organic layers were washed with water (4x50ml), then saturated brine (2x30ml), dried over MgSC>4 and passed through an 1ST Phase Separator. Evaporation gave a residue which was purified by HPLC preparative chromatography to give the title compound (5.4 mg); m/z: 403.08 [M+1]; 1H NMR (500 MHz, DMSO-d6) δ ppm 2.43 (s, 3 H) 2.82 (s, 2 H) 5.35 (s, 3 H) 7.65 - 7.69 (m, 1 H) 7.80(d, J=3.97 Hz, 2 H) 8.09 (d, J=8.55 Hz, 1 H) 8.12 (d, J=1.53 Hz, 1 H) 8.32 (d, J=5.49 Hz, 1 H) 10.47 (s, 1 H). Compound 4: 8-chloro-1 -methyl-N-(2-methyl-4-pyridin yl)-4H-imidazor5, 1 - ciri ,41benzothiazine-3-carboxamide
Figure imgf000066_0001
To a solution of 2-methyl-4-pyridinamine (52.7 mg, 0.487 mmol) in 1 ,4-dioxane (1.5 mL) stirred under argon at room temp was added a solution of trimethylaluminium (0.244 mL, 0.487 mmol) in heptane dropwise over 5 min, and the reaction mixture was stirred at rt for 1 hr. To this reaction mixture was added a solution of ethyl 8- chloro-1 -methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 6) (47 mg, 0.122 mmol) in 1 ,4-dioxane (1.5 mL) dropwise at room temperature under argon. The resulting mixture was heated to 100 °C and stirred at that temperature for 30h. Dioxane/water (30/1 ) was added and the reaction mixture was evaporated to dryness. DCM (50 ml) was added and stirred for 30 minutes. Staurated aqueous sodium bicarbonate solution (20 ml) was added and stirring was continued for 15 min.. The layers were separated by passing through an 1ST Phase separator and the aqueous layer was extracted with DCM (3x50ml). The combined organic layers were washed with water (100 ml) and brine (100ml), dried over sodium sulphate and passed through an 1ST Phase separator. Evaporation gave a residue which was purified using a Biotage SP1 , Snap Si 25 g, 25 ml/min in the gradient of 2N
NH3/MeOH in DCM: 0% of 2N NH3/MeOH for 1 CV then from 0-8% in 25 CV to give the title compound (28 mg); m/z: 371 [M+1]; 1H NMR (500 MHz, DMSO-d6) d ppm 2.42 (s, 3 H) 2.70 (s, 3 H) 4.44 (s, 2 H) 7.44 (dd, J=8.54, 2.14 Hz, 1 H) 7.67 (d,
J=8.24 Hz, 2 H) 7.79 (s, 1 H) 7.95 (d, J=2.14 Hz, 1 H) 8.30 (d, J=5.49 Hz, 1 H) 10.25
(s, 1 H).
Compound 5: 8-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5,1- cl[1 ,4lbenzothiazine-3-carboxamide
Figure imgf000066_0002
To a solution of 2-chloro-4-pyridinamine (64.0 mg, 0.497 mmol) in 1 ,4-dioxane (1.5 mL) stirred under argon at room temperature was added a solution of
trimethylaluminium (0.249 mL, 0.497 mmol) in heptane dropwise over 5 min and the reaction mixture was stirred at room temperature for 1 hr. Then a solution of ethyl 8- chloro-1 -methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 6) (48 mg, 0.124 mmol) in 1 ,4-dioxane (1.5 mL) was added dropwise at rt under argon. The resulting mixture was heated to 90 °C and stirred at that temperature for 25 hours. The reaction mixture was cooled to 0°C and quenched with dioxane/water (30/1 ). The entire reaction mixture was then evaporated to dryness. DCM (50 ml) was added and stirred for 0.5 h. Saturated sodium bicarbonate solution (20 ml) was added and stirring was continued for 15 min. The layers were separated by passing through an 1ST Phase separator and the aqueous layer was extracted with DCM (3x50ml). The combined organic layers were washed with water (100 ml) and brine (100ml), dried over sodium sulphate and passed through an 1ST Phase separator. Evaporation gave a residue which was purified on a Biotage SP1 Snap Si 25g, 25 ml/min.; in the gradient of 2N NH3/MeOH in DCM 1 % of 2N NH3/MeOH for 1 CV then from 1-8% in 30 CV to give an impure product. This product was washed with DCM/MeOH(10/1 ) and then with diisopropyl ether to give the title compound (10.2 mg) The combined washings were purified using a Biotage SP1 Snap Si 25g, 25 ml/min eluting with a gradient of EtOAc in Cyclohexane 40% of EtOAc for 1 CV then from 40-100% in 20 CV to give the title compound; m/z: 391 [M+1]; 1H NMR (500 MHz, DMSO-d6) d ppm 2.71 (s, 3 H) 4.44 (s, 2 H) 7.44 (dd, J=8.39, 1.98 Hz, 1 H) 7.68 (d, J=8.24 Hz, 1 H) 7.90 (dd, J=5.65, 1.68 Hz, 1 H) 7.95 (d, J=2.14 Hz, 1 H) 8.09 (d, J=1.53 Hz, 1 H) 8.27 (d, J=5.49 Hz, 1 H) 10.69 (s, 1 H).
Compound 6: 8-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazor5,1- clH ,41benzothiazine-3-carboxamide 5,5-dioxide
Figure imgf000067_0001
To a stirred solution of 8-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5,1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 5) (13 mg, 0.033 mmol) in dichloromethane (DCM) (1 ml) and methanol (300 μΙ) was added m-CPBA (17.2 mg, 0.1 mmol) dropwise in methanol (300 μΙ) over 5 min, and the mixture was allowed to warm to room temperature and stirring was continued for 18h. Further m-CPBA (5.73 mg, 0.033 mmol) in methanol (300 μΙ) was added then DCM (1 .5 ml) then MeOH (400 ul) added to dissolved the resulting suspension. The mixture was stirred for 12h. Water was added (10ml) and the mixture was extracted with DCM (3x10ml). The combined organic extracts were washed with NaHCC>3(sat. sol) (4x25ml) dried by passing through an 1ST Phase Separator and evaporated. The resulting residue was purified on Biotage SP1 Snap Si 25g in the gradient of 2N NH3/MeOH in DCM 2% for 1 CV then from 2-50% in 40 CV to give the title compoundafter wahing with dry methanol (2.9 mg); m/z: 423 [M+1 ]; 1H NMR (300 MHz, DMSO-d6) d ppm 2.83 (s, 1 1 H) 5.35 (s, 7 H) 7.81 (dd, J=8.38, 1 .60 Hz, 4 H) 7.90 (dd, J=5.65,1 .70 Hz, 4 H) 8.02 - 8.15 (m, 1 1 H) 8.30 (d, J=5.65 Hz, 4 H) 10.88 (s, 4 H).
Compound 7: 8-Chloro-/V-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4/-/-imidazo[5,1 - cl[1 ,4lbenzoxazine-3-carboxamide
Figure imgf000068_0001
To a solution of 2-chloro-4-pyridinamine (51 .3 mg, 0.399 mmol) in dry 1 ,4-dioxane (1 .25 mL) at room temperature was added trimethylaluminium (0.2 mL, 0.399 mmol) dropwise under argon and the solution stirred at room temperature for 1 hour. Ethyl 8-chloro-7-fluoro-1-methyl-4/-/-imidazo[5,1-c][1 ,4]benzoxazine-3-carboxylate
(intermediate 14, 31 mg, 0.100 mmol) in dry 1 ,4-dioxane (1 .25 mL) was added, and the mixture was heated at 100 °C for 20 hours. On cooling to room temperature, the reaction mixture was quenched with water (2 mL), stirred for 10 minutes and concentrated under vacuum. The residue was diluted with DCM (50 mL) and washed with 0.5 M NaOH (3x50 mL). The combined organic extracts were diluted with water (50 mL) and the pH adjusted to 2 with 0.1 M HCI. The layers were separated and the aqueous layer extracted with DCM (50 mL). The combined organic extracts were washed with saturated aqueous sodium bicarbonate solution (50 mL), brine (50 mL), and dried over Na2S04. The solvent was removed under vacuum and the crude product was triturated with EtOAc to give the title compound (21.5 mg); m/z (ES): 393.13 [M+H]+; 1H NMR (500 MHz, DMSO-d6) d ppm: 2.77 (s, 3 H), 5.53 (s, 2 H), 7.47 (d, J=10.07 Hz, 1 H), 7.87 (dd, J=5.49, 1.83 Hz, 1 H), 7.99 (d, J=7.32 Hz, 1 H), 8.06 (d, J=1.83 Hz, 1 H), 8.26 (d, J=5.80 Hz, 1 H), 10.75 (s, 1 H). Compound 8: 8-Chloro-/V-(2-chloro-4-pyridinyl)-7-fluoro-4H-ri ,2,31triazolor5,1- cl[1 ,4lbenzoxazine-3-carboxamide
Figure imgf000069_0001
To a solution of 2-chloro-4-pyridinamine (37.8 mg, 0.294 mmol) in dry 1 ,4-dioxane (1 mL) at room temperature was added trimethylaluminium (0.147 mL, 0.294 mmol) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 8-chloro-7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzoxazine-3-carboxylate (intermediate 15, 21 .9 mg, 0.074 mmol) in dry 1 ,4-dioxane (1 mL) was added and the reaction mixture was heated at 100 °C for 20 hours. The reaction mixture was cooled to room temperature and carefully quenched with water (2 mL). The mixture was stirred for 10 minutes and concentrated under vacuum. The residue was diluted with DCM (25 mL) and washed with 0.5 M NaOH (3x15 mL). The combined organic extracts were diluted with water (30 mL) and the pH adjusted to 2 with 0.1 M HCI. The layers were separated and the aqueous layer was extracted with DCM (25 mL). The combined organic extracts were washed with saturated aqueous sodium bicarbonate (30 mL), brine (30 mL) and dried over Na2S04. The solvent was removed under vacuum and the residue was triturated with EtOAc to give the title compound (12.7 mg); m/z (ES): 380.10 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 5.83 (s, 2 H), 7.50 (d, J=9.77 Hz, 1 H), 7.86 (dd, J=5.80, 1.83 Hz, 1 H), 8.03 (d, J=1.53 Hz, 1 H), 8.22 - 8.37 (m, 2 H), 1 1.39 (s, 1 H).
Compound 9: N-(2-Chloro-4-pyridinyl)-5,6-dihvdro-4H-ri ,2,31triazolon ,5- aimbenzazepine-3-carboxamide
Figure imgf000069_0002
To a solution of 2-chloro-4-pyridinamine (80 mg, 0.622 mmol) in dry 1 ,4-dioxane (2 mL) at room temperature was added trimethylaluminium (0.31 1 mL, 0.622 mmol) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 5,6-dihydro-4H-[1 ,2,3]triazolo[1 ,5-a][1]benzazepine-3-carboxylate (intermediate 20, 40 mg, 0.155 mmol) in dry 1 ,4-dioxane (2 mL) was added and the reaction mixture was heated at 100 °C for 20 hours. The reaction mixture was cooled to room temperature and carefully quenched with water (3 mL), stirred for 10 minutes and concentrated under vacuum. The residue was diluted with DCM (50 mL) and washed with 0.5 M NaOH (3x30 mL). The combined organic extracts were diluted with water (50 mL) and the pH adjusted to 2.0 with 5 M HCI. The layers were separated and the aqueous layer extracted with DCM (50 mL). Combined organic extracts were washed with brine (50 mL) and dried over Na2S04. The solvent was removed under vacuum and the crude product was purified on a FlashMaster II Instrument (LC-Si cartridge 2 g) eluting with an n-hexane/EtOAc gradient starting from 100/0 v/v for 5 minutes going to 70/30 v/v in the next 45 minutes to give the title compound (41 mg); m/z
(ES): 340.13 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm: 2.26 (quin, J=7.10 Hz, 2 H), 2.58 (t, J=7.02 Hz, 2 H), 3.08 (t, J=7.17 Hz, 2 H), 7.52 - 7.60 (m, 3 H), 7.73 - 7.79 (m, 1 H), 7.91 (dd, J=5.65, 1.68 Hz, 1 H), 8.08 (d, J=1.53 Hz, 1 H), 8.32 (d, J=5.49 Hz, 1 H), 1 1 .24 (s, 1 H).
Compound 10: N-(2-Chloro-4-pyridinyl)-1 -methyl-5,6-dihydro-4H-imidazo[1 ,5- aimbenzazepine-3-carboxamide
Figure imgf000070_0001
To a solution of 2-chloro-4-pyridinamine (1 14 mg, 0.888 mmol) in dry 1 ,4-dioxane (3 mL) at room temperature was added trimethylaluminium (0.444 mL, 0.888 mmol) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 1 -methyl-5,6-dihydro-4H-imidazo[1 ,5-a][1 ]benzazepine-3-carboxylate
(intermediate 21 , 60 mg, 0.222 mmol) in dry 1 ,4-dioxane (3 mL) was added and the reaction mixture was cooled to room temperature. The reaction was carefully quenched with water (5 mL), stirred for 10 minutes and concentrated under vacuum. The residue was diluted with DCM (50 mL) and washed with 0.5 M NaOH (3x50 mL). The combined organic extracts were diluted with water (50 mL) and the pH adjusted to 2.0 with 5 M HCI. The layers were separated and aqueous layer extracted with DCM (20 mL). The combined organic extracts were dried over Na2S04. The solvent was removed under vacuum and the residue was purified on a FlashMaster II
Instrument (LC-Si cartridge 2 g) eluting with an n-hexane/EtOAc gradient starting from 100/0 v/v for 5 minutes going to 70/30 v/v over 45 minutes to obtain the title compound (22.1 mg); m/z (ES): 353.12 [M+H]+; 1H NMR (600 MHz, DMSO-d6) d ppm: 1.97 - 2.12 (m, 3 H), 2.25 - 2.33 (m, 1 H), 2.42 (s, 3 H), 2.68 (dd, J=13.52, 6.02 Hz, 1 H), 3.66 - 3.73 (m, 1 H), 7.41 - 7.50 (m, 3 H), 7.54 - 7.58 (m, 1 H), 7.88 (dd, J=5.58, 1.92 Hz, 1 H), 8.09 (d, J=1.74 Hz, 1 H), 8.24 (d, J=5.58 Hz, 1 H), 10.
Compound 1 1 : N-(2-chloro-4-pyridinyl)-1 -methyl-4H-imidazor5,1 - ciri ,41benzothiazine-3-carboxamide
Figure imgf000071_0001
To a solution of 2-chloro-4-pyridinamine (131 mg, 1 .021 mmol) in dry 1 ,4-dioxane
(2.5 mL) at room temperature was added trimethylaluminum (0.510 mL, 1.021 mmol) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 1 -methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3-carboxylate (intermediate 25) (70 mg, 0.255 mmol) in dry 1 ,4-dioxane (2.5mL) was added and the reaction mixture was heated at 80 °C for 20 h. On cooling the reaction mixture was quenched with dioxane/water (30/1 mL) and stirred for 10 min. The resulting precipitate was decanted and the solvent evaporated. The precipitate was dissolved in DCM and washed with water. The aqueous phase was extracted with DCM (4 x 25 ml). The combined organic extracts were washed with sat NaCI (25 ml), and dried over anhydrous IS^SC^. The mixture was filtered using a phase separator filter tube and the solvent was removed in vacuo. Attempted purification of the residue using a Biotage SP1 purification system using Normal phase Silica SNAP 25 g column eluting with a gradient of MeOH (2N NH3) in DCM (0 % in 1 CV, 0-10 % in 20 CV), failed to isolate the titled product. The fractions were combined, the solvent evaporated and the residue purified using a Biotage SP1 purification system using Normal phase Silica SNAP 10 g column eluting with a gradient of EtOAc in Cy (3-66 %, 12 CV, 66-100 % 10 CV) to give the title compound (44.5 mg); m/z (ES): 357.1 [M+H]+; 1H NMR (300 MHz, DMSO-d6) d ppm 2.68 (s, 3 H) 4.42 (s, 2 H) 7.31 - 7.40 (m, 1 H) 7.42 - 7.50 (m, 1 H) 7.61 - 7.67 (m, 1 H) 7.81 - 7.93 (m, 2 H) 8.06 -8.1 1 (m, 1 H) 8.24 - 8.29 (m, 1 H) 10.66 (s, 1 H).
Compound 12: 1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazor5,1 - ciri ,41benzothiazine-3-carboxamide
Figure imgf000072_0001
To a solution of 2-methyl-4-pyridinamine (1 10 mg, 1 .021 mmol) in dry 1 ,4-dioxane (2.5 mL) at room temperature was added trimethylaluminum (0.510 mL, 1.021 mmol) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 1 -methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3-carboxylate (intermediate 25) (70 mg, 0.255 mmol) in dry 1 ,4-dioxane (2.5mL) was added and the reaction mixture was heated at 80 °C for 20 h. The temperature was raised to 95 °C and left stirring for a further 6 hours. The reaction mixture was cooled to 0 °C and quenched with dioxane/water (30/1 mL) and stirred for 10 min.The resulting precipitate was decanted and the solvent was evaporated. The solid was dissolved in DCM and water and extracted with DCM. Reaction mixture was dissolved in saturated sodium bicarbonate solution and extracted with DCM (4 x 25 ml). The combined organic extracts were washed with sat NaCI (25 ml), and dried over anhydrous IS^SC^. Solvent was filtered over phase separator filter tube and removed in vacuo to give a crude product as an yellow solid. The residue was purified via Biotage SP1 purification system using Normal phase Silica SNAP 25 g column in the gradient of MeOH (2N NH3) in DCM (1 % in 3 CV, 1 -2 % in 8 CV, 2-3 % in 4 CV) to give the title compound (47.9 mg); m/z (ES): 337.2 [M+H]+; 1H NMR (500 MHz, CHLOROFORM- d) d ppm 2.56 (s, 3 H) 2.68 (s, 3 H) 4.40 (s, 2 H) 7.27 - 7.30 (m, 1 H) 7.34 - 7.38 (m, 1 H) 7.40 - 7.42 (m, 1 H) 7.51 -7.54 (m, 1 H) 7.55 - 7.58 (m, 2 H) 8.40 - 8.42 (m, 1 H) 9.05 (s, 1 H)
Compound 13: N-(2-chloro-4-pyridinyl)-1 -methyl-4H-imidazo[5,1 - clH ,41benzothiazine-3-carboxamide 5,5-dioxide
Figure imgf000072_0002
To a solution of N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5,1- c][1 ,4]benzothiazine-3-carboxamide (Compound 1 1 , 34 mg, 0.095 mmol) in a mixture of dry dichloromethane (DCM) (1 .45 ml) and dry methanol (483 μί) was added mCPBA (37.8 mg, 0.219 mmol) and the mixture was stirred at rt for 1 day. The reaction mixture was diluted with DCM (20 mL) and washed with water (20 mL) and saturated aquesous sodium bicarbonate (20 mL). The combined aqueous layers were extracted with DCM (4x15 ml_). The combined organic layers were dried over Na2SC> , filtered and evaporated. The residue was purified via Biotage SP1 purification system using Normal phase Silica SNAP 10 g column using a gradient of EtOAc in Cy (30-100 % in 25 CV) to give the title compound (23 mg); m/z (ES):
389.01 [M+H]+; 1 H NMR (500 MHz, DMSO-d6) d ppm 2.80 (s, 3 H) 5.33 (s, 2 H) 7.71 - 7.75 (m, 1 H) 7.90 - 7.92 (m, 1 H) 7.95 - 8.00 (m, 1 H) 8.02 - 8.05 (m, 1 H) 8.07 - 8.10 (m, 2 H) 8.29 - 8.31 (m, 1 H) 10.87 (s, 1 H).
Compound 14: N-(6-chloro-2-pyridinyl)-1 -methyl-4H-imidazor5,1 - clH ,41benzothiazine-3-carboxamide 5,5-dioxide
Figure imgf000073_0001
To a solution of N-(6-chloro-2-pyridinyl)-1-methyl-4H-imidazo[5,1- c][1 ,4]benzothiazine-3-carboxamide (intermediate 2.669 g, 0.193 mmol) in a mixture of dry dichloromethane (5 mL) and dry methanol (2 mL) was added mCPBA (77 mg, 0.445 mmol) and the mixture was stirred for 5 h. Further mCPBA (33.4 mg, 0.193 mmol) was added and the mixture was stirred at room temperature overnight. The reaction mixture was diluted with DCM (20 mL) and washed with water (20 mL) and saturated aqueous sodium bicarbonate solution (20 mL). The combined water aqueous layers were extracted with DCM (4x20 mL) and the combined organic extracts were dried over Na2SC> , filtered and evaporated. The residue was triturated with methanol to obtain the title compound (63 mg); m/z (ES): 388.96 [M+H]+; 1H NMR (500 MHz, DMSO-d6) d ppm 2.79 (s, 3 H) 5.32 (s, 2 H) 7.30 (d, 1 H) 7.73 (t, 1 H) 7.92 - 8.00 (m, 2 H) 8.03 (d, 1 H) 8.09 (d, 1 H) 8.16 (d, 1 H)9.74 (s, 1 H). Compound 15: N-(2-chloro-4-pyridinyl)-4H-ri,2,3ltriazolor5,1-cin,4lbenzothiazine-3- carboxamide
Figure imgf000073_0002
To a solution of 2-chloro-4-pyridinamine (197 mg, 1 .531 mmol) in dry 1 ,4-dioxane (3 mL) at room temperature was added dropwise trimethylaluminum (0.765 mL, 1.531 mmol) under argon and the solution was stirred at room temperature for 1 hour. Ethyl 4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxylate (intermediate 27, 100 mg, 0.383 mmol) in dry 1 ,4-dioxane (3 mL) was added and the reaction mixture was heated at 100 °C overnight. The reaction mixture was cooled to 0 °C and quenched with dioxane/water (30ml_/1 mL) and stirred for 10 min. The reaction mixture was concentrated, dissolved in water (25 mL) and extracted with DCM (4x25 ml). The combined organic extracts were washed with brine (25 ml) and dried over anhydrous Na2SC>4. The solvent was filtered through a phase separator filter tube and removed in vacuo. The residue was purified using a Biotage SP1 purification system using a Normal phase Silica SNAP 25 g column eluting with a gradient of EtOAc in cyclohexane (3 % in 1 CV, 3-66 % in 16 CV, 66-80 % in 5 CV) to give a solid which was recrystalised from EtOAc:Cyclohexane (1 :10) to obtain the title compound (31 mg); m/z (ES): 344.02 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 4.67 (s, 2 H) 7.44 - 7.52 (m, 2 H) 7.65 (dd, 1 H) 7.90 (dd, 1 H) 8.06 (s, 1 H) 8.16 (dd, 1 H) 8.33 (d, 1 H) 1 1.34 (s, 1 H). Compound 16: N-(2-chloro-4-pyridinyl)-4H-ri ,2,3ltriazolor5,1-ciri,4lbenzothiazine-3- carboxamide 5,5-dioxide
Figure imgf000074_0001
To a solution of 2-chloro-4-pyridinamine (131 mg, 1.023 mmol) in dry 1 ,4-dioxane (2 mL) at room temperature was added trimethylaluminum (0.51 1 mL, 1.023 mmol) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3-carboxylate 5,5-dioxide (75 mg, 0.256 mmol) in dry 1 ,4-dioxane (2 mL) was added and the reaction mixture was heated at 92 °C overnight. The mixture was cooled to 0 °C and quenched with dioxane/water (30 mL/1 mL) and stirred for 10 min. The mixture was concentrated and dissolved in water (25 mL) and extracted with DCM (4 x 35 ml). The combined organic extracts were washed with 1 N HCI (3x25 ml), then with brine (25 ml) and dried over anhydrous Na2SC> . The solvent was filtered through a phase separator filter tube and removed in vacuo. The residue was triturated with MeOH to obtain the title compound (24 mg); m/z (ES): 375.90 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 5.58 (s, 2 H) 7.83 - 7.90 (m, 2 H) 8.03 - 8.08 (m, 2 H) 8.15 (dd, J=7.93, 1.22 Hz, 1 H) 8.36 (d, J=5.80 Hz, 1 H) 8.44(d, J=7.32 Hz, 1 H) 1 1.50 (s, 1 H). Compound 17: N-(2-methyl-4-pyridinyl)-4H-[1 ,2,3ltriazolo[5,1 -cl[1 ,4lbenzothiazine-3- carboxamide
Figure imgf000075_0001
To a solution of 2-methyl-4-pyridinamine (166 mg, 1 .531 mmol) in dry 1 ,4-dioxane (3 mL) at room temperature was added trimethylaluminum (0.765 ml_, 1.531 mmol) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 27, 100 mg, 0.383 mmol) in dry 1 ,4-dioxane (3 mL) was added and the reaction mixture was heated at 90 °C overnight. The mixture was cooled to 0 °C and quenched with dioxane/water (30/1 mL) and stirred for 10 min. The reaction mixture was
concentrated and dissolved in water (25 mL) and extracted with DCM (4 x 25 ml). The combined organic extracts were washed with brine (25 ml), and dried over anhydrous IS^SC^. The solvent was filtered through a phase separator filter tube and evaporated in vacuo. The resulting residue was purified using a Biotage SP1 purification system using a Normal phase Silica SNAP 25 g column, eluting with a the gradient of EtOAc in Cyclohexane (12 % in 1 CV, 12-100 % in 15 CV, 100 % in 2 CV) to give the title compound (15 mg); m/z (ES): 324.06 [M+H]+; 1H NMR (500 MHz, DMSO-d6) d ppm 2.44 (s, 3 H) 4.66 (s, 2 H) 7.43 - 7.52 (m, 2 H) 7.65 (d, 1 H) 7.68 (d, 1 H) 7.79 (s, 1 H) 8.16 (d, 1 H) 8.35 (d, 1 H)10.98 (s, 1 H).
Compound 18: N-(2-chloro-4-pyridinyl)-1 -(trifluoromethyl)-4H-imidazor5,1- ciri ,41benzothiazine-3-carboxamide
Figure imgf000075_0002
To a solution of 2-chloro-4-pyridinamine (15.66 mg, 0.122 mmol) in dry 1 ,4-dioxane (300 μί) at room temperature was added trimethylaluminum (60.9 μί, 0.122 mmol) dropwise under Argon and the solution was stirred at room temperature for 1 hour. Ethyl 1 -(trif luoromethyl)-4H-imidazo[5, 1 -c][1 ,4]benzothiazine-3-carboxylate
(intermediate 30, 10 mg, 0.030 mmol) in dry 1 ,4-dioxane (300 μί) was added and the reaction mixture was heated 80 °C for 5 hours. The reaction mixture was cooled to room temperature and quenched with dioxane/water (1 .0/0.4 mL) and stirred for 10 min. Water (15 mL) was added and the mixture was extracted with DCM (4x15 ml). The combined organic extracts were washed with 1 N HCI (4x10 mL) and brine (15 ml) and dried over anhydrous IS^SC^. The solvent was filtered over a phase separator filter tube and evaporated in vacuo. The resulting residue was purified using a Biotage SP1 purification system using a Normal phase Silica SNAP 10 g column in a gradient of EtOAc in Cyclohexane (5 % in 1 CV, 5-66 % in 15 CV, 66 % in 5 CV) to give the title compound (9.7 mg); m/z (ES): 41 1.15 [M+H]+; 1H NMR (500 MHz, DMSO-d6) d ppm 4.39 (s, 2 H) 7.34 - 7.39 (m, 1 H) 7.40 - 7.46 (m, 1 H) 7.57 - 7.60 (m, 1 H) 7.62 - 7.66 (m, 1 H) 7.77 - 7.80 (m, 1 H) 7.95 - 7.98 (m, 1 H) 8.20 - 8.22 (m, 1 H) 10.70 (s, 1 H).
Compound 19: N-(2-chloro-4-pyridinyl)-1 -cvclopropyl-4H-imidazor5, 1 - cl[1 ,4lbenzothiazine-3-carboxamide
Figure imgf000076_0001
To a solution of 2-chloro-4-pyridinamine (34.2 mg, 0.266 mmol) in dry 1 ,4-dioxane (500 μΙ_) at room temperature was added trimethylaluminum (133 μΙ_, 0.266 mmol) dropwise under argon and the solution was stirred at room temperarture for 1 hour. Ethyl 1 -cyclopropyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 31 , 20 mg, 0.067 mmol) in dry 1 ,4-dioxane (500 μΙ_) was added and the reaction mixture was heated at 80 °C overnight. The reaction mixture was cooled to room temperature, quenched with dioxane/water (2.0/0.5 mL) and stirred for 10 min. 1 M NaOH (15 mL) was added and the mixture was extracted with DCM (4x15 ml). The combined organic extracts were washed with 1 N HCI (4x10 mL) then brine (15 ml), and dried over anhydrous Na2SC> . The mixture was filtered through a phase separator filter tube and the solvent removed in vacuo. The resulting residue was purified using a Biotage SP1 purification system using a Normal phase Silica SNAP 10 g column eluting with a gradient of EtOAc in Cyclohexane (5 % in 1 CV, 5-30 % in 10 CV, 30-66 % in 10 CV) to give (18.5 mg); m/z (ES): 383.17 [M+H]+; 1H NMR (300 MHz, DMSO-d6) d ppm 1 .07 - 1.16 (m, 2 H) 1.17 - 1.24 (m, 2 H) 2.15 - 2.26 (m, 1 H) 4.40 (s, 2 H) 7.33 - 7.40 (m, 1 H) 7.43 - 7.52 (m, 1 H) 7.63 - 7.69 (m, 1 H) 7.85 - 7.91 (m, 1 H) 8.04 - 8.07 (m, 1 H) 8.07 - 8.13 (m, 1 H) 8.25 - 8.31 (m, 1 H) 10.22 - 10.28 (m, 1 H). Compound 20: N-(2-chloro-4-pyridinyl)-7-fluoro-4H-ri,2,3ltriazolor5,1 - cl[1 ,4lbenzoxazine-3-carboxamide
Figure imgf000077_0001
To a solution of 2-chloro-4-pyridinamine (137 mg, 1 .064 mmol) in dry 1 ,4-dioxane (2.5 mL) at room temperature was added trimethylaluminum (0.532 mL, 1.064 mmol) (2M in heptanes) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 7-fluoro-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzoxazine-3- carboxylate (intermediate 37, 70 mg, 0.266 mmol) in dry 1 ,4-dioxane (2.5 mL) was added and the reaction mixture was heated at 90 °C overnight. The reaction mixture was cooled to room temperature and quenched with dioxane/water (5.0/1.0 mL) and stirred for 10 min. The mixture was evaporated and the residue was dissolved in saturated aqueous sodium bicarbonate solution (40 mL). The mixture was extracted with DCM (4 x 30 ml) and the combined organic extracts were washed with 1 N HCI (4x25 mL), then brine (20 ml) and dried over anhydrous IS^SC^. The mixture was filtered through a phase separator filter tube and the solvent removed in vacuo. The residue was purified using a Biotage SP1 purification system using a Normal phase Silica SNAP 10 g column, eluting with a gradient of EtOAc in Cyclohexane (8 % in 1 CV, 8-66 % in 15 CV, 66 % in 5 CV), followed by precipitation from
cyclohexane: EtOAc (10:1 ) to givethe title compound (58 mg); m/z (ES): 346.05
[M+H]+; 1H NMR (500 MHz, DMSO-d6) d ppm 5.82 (s, 2 H) 7.06 - 7.12 (m, 1 H) 7.21 - 7.27 (m, 1 H) 7.85 - 7.90 (m, 1 H) 8.03 - 8.07 (m, 1 H) 8.08 - 8.14 (m, 1 H) 8.28 - 8.36 (m, 1 H) 1 1.38 (s, 1 H).
Compound 21 : N-(6-chloro-2-pyridinylWluoro-4H-M .2,3ltriazolor5,1 - ciri,41benzoxazine-3-carboxamide
Figure imgf000077_0002
To a solution of 6-chloro-2-pyridinamine (68.4 mg, 0.532 mmol) in dry 1 ,4-dioxane (1.5 mL) at room temperature was added trimethylaluminum (0.266 mL, 0.532 mmol) (2M in heptanes) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 7-fluoro-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzoxazine-3- carboxylate (intermediate 37, 35 mg, 0.133 mmol) in dry 1 ,4-dioxane (1.5 mL) was added and the reaction mixture was heated at 90 °C with stirring for 3 hours. The reaction mixture was cooled to room temperature and quenched with dioxane/water (5.0/1.0 mL) and stirred for 10 min. The solvent was evaporated, dissolved in 1 N NaOH (40 ml) and extracted with DCM (4x30 ml). The combined organic extracts were washed with 1 N HCI (4x25 mL) then brine (20 ml) and dried over anhydrous Na2SC>4. The mixture was filtered through a phase separator filter tube and the solvent removed in vacuo. A mixture a cyclohexane/ethyl acetate (10:1 ) was added to the resulting residue causing precipitation of the title compound (35.4 mg); m/z (ES): 346.30 [M+H]+; 1 H NMR (500 MHz, DMSO-d6) d ppm 5.82 (s, 2 H) 7.08 - 7.14 (m, 1 H) 7.22 - 7.27 (m, 1 H) 7.30 - 7.36 (m, 1 H) 7.92 - 7.97 (m, 1 H) 8.08 - 8.14 (m, 2 H) 10.59 - 10.65 (m, 1 H).
Compound 22: N-(2-chloro-4-pyridinyl)-7-fluoro-1 -methyl-4H-imidazo[5,1 - cl[1 ,4lbenzoxazine-3-carboxamide
Figure imgf000078_0001
To a solution of 2-chloro-4-pyridinamine (84 mg, 0.652 mmol) in dry 1 ,4-dioxane (2 mL) at room temperature was added trimethylaluminum (0.326 mL, 0.652 mmol) (2M in heptanes) dropwise under argon and the solution was stirred at room temperature for 1 hour. Ethyl 7-fluoro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzoxazine-3-carboxylate (intermediate 38, 45 mg, 0.163 mmol) in dry 1 ,4-dioxane (2 mL) was added and the reaction mixture was heated at 90 °C with stirring for 5 h. The reaction mixture was cooled to room temperature and quenched with dioxane/water (5.0/1.0 mL) and stirred for 10 min. The solvent was evaporated and the residue was dissolved in 1 N NaOH (40 ml) and extracted with DCM (4 x 30 ml). Combined organic extracts were washed with 1 N HCI (4x25 mL) then brine (20 ml) and dried over anhydrous
Na2SC> . The mixture was filtered through a phase separator filter tube and the solvent was removed in vacuo. A mixture a cyclohexane/ethyl acetate (10:1 ) was added to the resulting residue causing precipitation of the title compound (51 mg); m/z (ES): 359.05 [M+H]+; 1H NMR (500 MHz, DMSO-d6) d ppm 2.76 (s, 3 H) 5.53 (s, 2 H) 7.06 - 7.1 1 (m, 1 H) 7.19 - 7.23 (m, 1 H) 7.84 - 7.91 (m, 2 H) 8.06 - 8.09 (m, 1 H) 8.25 - 8.28 (m, 1 H) 10.74 (s, 1 H).
Compound 23: 7-chloro-N-(2-chloro-4-pyridinvn-4H-M .2.3ltriazolor5.1- ciri ,41benzothiazine-3-carboxamide
Figure imgf000079_0001
To a solution of 2-chloro-4-pyridinamine (139 mg, 1 .082 mmol) in dry 1 ,4-dioxane (3 mL) stirred at room temperature under argon was added trimethylaluminium (2 M solution in heptanes) (0.541 mL, 1.082 mmol) dropwise and the reaction mixture was stirred for 1 hour. Ethyl 7-chloro-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3- carboxylate (intermediate 45, 80 mg, 0.271 mmol) in dry 1 ,4-dioxane (3 mL) was added and the reaction mixture was heated at 100 °C for 20 hours. The reaction mixture was cooled to room temperature and further trimethylaluminium (2 M solution in heptanes) (0.271 mL, 0.541 mmol) was added and heated at 100 °C with stirring for 25 hours. The mixture was cooled to room temperature and quenched with 20 ml of water. 50 ml of DCM was added and the resulting sludge was filtered through a paper, washing the solid with 80 ml of methanol. The combined filtrate and washings were concentrated under reduced pressure and the residue was dissolved in 50 ml of DCM. This solution was washed with water (2x100 ml) and the organic phase was separated using a phase separator cartridge. The solvent was evaporated and the residue was purified by chromatography eluting with a cyclohexane/EtOAc solvent system on normal phase SNAP coloumn (25 g) to give a product. The product was precipitated from dimethyl sulfoxide and preciptate. The precipitate was dissolved in DCM/Methanol and the solvent were evaporated under reduced pressure to give the title compound (8.4 mg); m/z (ES): 377.90 [M+H]+; 1H NMR (500 MHz,
CHLOROFORM-d) δ ppm 4.55 (s, 2 H) 7.39 (dd, J=8.70, 2.14 Hz, 1 H) 7.48 - 7.53 (m, 2 H) 7.84 (d, J=1 .68 Hz, 1 H) 8.13 (d, J=8.70 Hz, 1 H) 8.35 (d, J=5.49 Hz, 1 H) 9.10 (s, 1 H). Compound 24: 7-chloro-N-(2-methyl-4-pyridinyl)-4H-ri ,2,3ltriazolor5,1- cl[1 ,4lbenzothiazine-3-carboxamide
Figure imgf000079_0002
To a solution of 2-methyl-4-pyridinamine (1 17 mg, 1 .082 mmol) in dry 1 ,4-Dioxane, (3 mL) stirred at room temperature under argon was added trimethylaluminium (2 M solution in heptanes) (0.541 ml_, 1.082 mmol) dropwise and the reaction mixture was stirred for 1 hour. Ethyl 7-chloro-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3- carboxylate (intermediate 45, 80 mg, 0.271 mmol) in dry 1 ,4-dioxane (3 mL) was added dropwise and the reaction mixture was heated at 100 °C for 20 hours. On cooling to room temperature, 20 ml of water was added and the resulting slurry was filtered through a filter paper, washing with approximately 80 ml of methanol. The combined filtrate and washings were evaporated and to the residue were added 50 ml of water and 50 ml of DCM and residue was dissolved using an ultrasound bath. The layers were separated using a phase separator cartridge, and the aqueous layer was extracted with 50 ml of DCM. The combined organic layers were evaporated and the resulting residue was purified by chromatography using a Biotage SP1 purifiation system using 10 g normal phase silica SNAP column eluting with DCM/Methanol (2N NH3) solvent system (gradient 1 -3 % of Methanol(2 N NH3) in 20 CV) to give the title compound (65 mg); m/z (ES): 358.08 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 2.58 (s, 3 H) 4.56 (s, 2 H) 7.38 (dd, J=8.62, 2.21 Hz, 1 H) 7.43 (dd, J=5.65, 1.98 Hz, 1 H) 7.50 (d, J=2.14 Hz, 1 H) 7.54 (d, J=1.83 Hz, 1 H) 8.13 (d, J=8.70 Hz, 1 H) 8.47 (d, J=5.65 Hz, 1 H) 8.99 (s, 1 H).
Compound 25: 7-chloro-N-(2-chloro-4-pyridinyl)-1 -methyl-4H-imidazo[5,1 - cl[1 ,4lbenzothiazine-3-carboxamide
Figure imgf000080_0001
To a solution of 2-chloro-4-pyridinamine (50 mg, 0.389 mmol) in dry 1 ,4-dioxane (1 .5 mL) stirred at room temperature under argon was added trimethylaluminium (2 M solution in heptanes) (0.194 mL, 0.389 mmol) dropwise and the reaction mixture was stirred for 1 hour. Ethyl 7-chloro-1-methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3- carboxylate (intermediate 46, 30 mg, 0.097 mmol) in dry 1 ,4-dioxane (1.5 mL) was added and heated at 100 °C for 20 hours. On cooling, 20 ml of water was added and the resulting slurry was filtered through a filter paper, and washing with aproximately 80 ml of methanol/DCM (1 :1 ) mixture. The combined filtrate and washings were evaporated under reduced pressure to dryness and the resulting residue was dissolved in 50 ml of DCM. The mixture was washed with 50 ml of water (x2) using a phase separator cartridge. The solvent from the DCM solution was evaporated and the resulting crude was purified by chromatography using Biotage SP1 purification system, 10 g normal phase silica SNAP column eluting with a cyclohexane/EtOAc solvent system (gradient 40-60 % of EtOAC in 20 CV) to give the title compound (20 mg); m/z (ES): 391 .04 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 2.66 (s, 3 H) 4.39 (s, 2 H) 7.34 (dd, J=8.70, 2.44 Hz, 1 H) 7.46 (d, J=8.39 Hz, 1 H) 7.51 (dd, J=5.65, 1.83 Hz, 1 H) 7.57 (d, J=2.29 Hz, 1 H) 7.80 (d, J=1.68 Hz, 1 H) 8.29 (d, J=5.65 Hz, 1 H) 9.15 (s, 1 H).
Compound 26: 7-chloro-N-(2-chloro-4-pyridinvn-4H-M .2.3ltriazolor5.1- clH ,41benzothiazine-3-carboxamide 5,5-dioxide
Figure imgf000081_0001
To a solution of 2-chloro-4-pyridinamine (39.2 mg, 0.305 mmol) in dry 1 ,4-dioxane (1 mL) stirred at room temperature under argon was added dropwise
trimethylaluminium (2 M solution in heptanes) (0.153 mL, 0.305 mmol) and the reaction mixture was stirred for 1 hour. Ethyl 7-chloro-4H-[1 ,2,3]triazolo[5,1- c][1 ,4]benzothiazine-3-carboxylate 5,5-dioxide (intermediate 47, 25 mg, 0.076 mmol) in dry 1 ,4-dioxane (1 mL) was added and the mixture was stirred at 100 °C for 20 hours. On cooling to room temperature, the mixture was quenched with 20 ml of water and the resulting slurry was filtered through a filter paper, washing through with aproximately 80 ml of methanol. The combined filtrate and washings were
evaporated and to the resulting residue was added 50 ml of water and 50 ml of DCM and the residue dissolved using an ultrasound bath. The layers were separated using a phase separator cartridge, and the aqueous layer extracted with 50 ml of DCM. The combined DCM phases were washed with 2x30 ml of 1 N HCI and the solvent removed. The residue was purified by chromatography on a Biotage SP1 purifiation system using 10 g normal phase silica SNAP column eluting with cyclohexane:EtOAc solvent system (gradient 15-50 % of EtOAc in 20 CV) to give a product that was triturated with diethy ether to give the title compound (1.2 mg); m/z (ES): 409.98
[M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 5.62 (s, 2 H) 7.88 (dd, J=5.65, 1.53 Hz, 1 H) 8.06 (d, J=1.37 Hz, 1 H) 8.12 (dd, J=8.77, 2.21 Hz, 1 H) 8.22 (d, J=2.14 Hz, 1 H) 8.36 (d, J=5.65 Hz, 1 H) 8.46 (d, J=8.70 Hz, 1 H) 1 1.52 (s, 1 H).
Compound 27: 7-chloro-1 -methyl-N-(2-methyl-4-pyridinyl)-4H-imidazor5,1 - ciri ,41benzothiazine-3-carboxamide
Figure imgf000082_0001
To a solution of 7-chloro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylic acid (24 mg, 0.085 mmol) in dry dichloromethane (8 mL) cooled in an ice/acetone mixture were added DIPEA (16 μΙ, 0.092 mmol) and isobutyl chloroformate (12.35 μΙ, 0.094 mmol) and the mixture was stirred for 24 h at room temperature. The solvent was evaporated under reduced pressure to give a residue (Residue A) which was stored under an Argon until needed in the next step. To a stirred solution of 2-methyl- 4-pyridinamine (37.0 mg, 0.342 mmol) in dry 1 ,4-dioxane (2 mL) under argon was added dropwise trimethylaluminium (2 M solution in hexanes) (171 μΙ, 0.342 mmol) and the mixture was stirred for 1 hour. To this mixture was added Residue A in dry 1 ,4-dioxane (2 mL). The resulting mixture was heated to 80 °C and was stirred for 3 hours. The mixture was diluted with 20 ml of water and basified with 1 N NaOH to pH 12 to 13. The mixture was extracted with 70 ml of DCM using phase separator cartridge and the organic extracts were evaporated to give a residue which was purified using a Biotage SP1 purification system and then by preparative liquid chromatography to give the title compound (1.3 mg); m/z (ES): 371.3 [M+H]+; 1H NMR (500 MHz, DMSO-d6) δ ppm 2.42 (s, 3 H) 2.67 (s, 3 H) 4.46 (s, 2 H) 7.51 (dd, J=8.85, 2.29 Hz, 1 H) 7.67 (dd, J=5.49, 1.22 Hz, 1 H) 7.74 - 7.81 (m, 2 H) 7.86 (d, J=8.85 Hz, 1 H) 8.30 (d, J=5.65 Hz, 1 H) 10.24 (s, 1 H).
Compound 28: 8-chloro-N-(2-chloro-4-Dyridinvn-7-fluoro-4H-n,2,3ltriazolor5,1 - ciri,41benzothiazine-3-carboxamide
Figure imgf000082_0002
To a solution of 2-chloro-4-pyridinamine (65.6 mg, 0.484 mmol) in dry 1 ,4-dioxane (2 mL) under argon was added dropwise trimethylaluminium (2M solution in heptanes) (0.242 mL, 0.484 mmol) and the mixture was stirred for 1 hour at room temperature. Ethyl 8-chloro-7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxylate (40 mg, 0.121 mmol) in dry 1 ,4-dioxane (2 mL) was added and the mixture was heated at 95 °C for 16 hours. On cooling to room temperature, 5 ml of water and 10 ml of DCM were added and the mixture was basified with 1 N NaOH. The layers were separated and the aqueous layer was extracted with 3x10 ml of DCM. The combined organic extracts were dried using phase separator cartridge and the solvent was evaporated. The resulting residue was purified using a Biotage SP1 purification device using 10 g normal phase silica SNAP column eluting with cyclohexane/EtOAc solvent system (gradient 5-35 % of EtOAc in 20 CV) to give the title compound (23 mg); m/z (ES): 396.2 [M+H]+; 1H NMR (500 MHz, DMSO-d6) d ppm 4.71 (s, 2 H) 7.87 - 7.93 (m, 2 H) 8.06 (d, J=1.68 Hz, 1 H) 8.31 - 8.37 (m, 2 H) 1 1 .37 (s, 1 H).
Compound 29: 8-chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-1 -methyl-4H-imidazor5,1- ciri ,41benzothiazine-3-carboxamide
Figure imgf000083_0001
To a solution of 2-chloro-4-pyridinamine (58.0 mg, 0.428 mmol) in dry 1 ,4-dioxane, Aldrich (2 mL) under argon was added trimethylaluminium (2 M in heptanes) (0.214 ml_, 0.428 mmol) and the mixture was stirred for 1 hour on room temperature. Ethyl 8-chloro-7-fluoro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate
(intermediate 55, 35 mg, 0.107 mmol) in dry 1 ,4-dioxane (2 mL) was added and the mixture was heated at 95 °C for 16 hours. The mixture was cooled to room temperature, 30 ml of water and 50 ml of DCM were added and the mixture was basified with 1 N NaOH. The layers were separated and the aqueous layer was extracted with 3x10 ml of DCM. The combined organic extracts were washed with 30 ml of 0.1 N HCI and the layers separated. The aqueous layer was washed with 30 ml of DCM in total and the combined organic layers were concentrated to give a residue which was triturated twice with diethyl ether to give the title compound (35.3 mg); m/z (ES): 409.3 [M+H]+; 1 H NMR (500 MHz, DMSO-d6) δ ppm 2.70 (s, 3 H) 4.47 (s, 2 H) 7.86 (d, J=9.16 Hz, 1 H) 7.90 (dd, J=5.72, 1.75 Hz, 1 H) 8.08 (d, J=1.53 Hz, 1 H) 8.12 (d, J=6.87 Hz, 1 H) 8.27 (d, J=5.65 Hz, 1 H) 10.69 (s, 1 H).
Compound 30: 8-chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-1 -methyl-4H-imidazor5,1- clH ,41benzothiazine-3-carboxamide 5,5-dioxide
Figure imgf000084_0001
To a solution of 2-chloro-4-pyridinamine (43.5 mg, 0.321 mmol) in dry 1 ,4-dioxane (2 mL) under argon was added dropwise trimethylaluminium (2M in heptanes) (0.161 mL, 0.321 mmol) and the mixture was stirred for 1 hour at room temperature. Ethyl 8- chloro-7-fluoro-1 -methyl-4H-imidazo[5, 1 -c][1 ,4]benzothiazine-3-carboxylate 5,5- dioxide (intermediate 56, 32 mg, 0.080 mmol) in dry 1 ,4-dioxane (2 mL) was added and the mixture was heated at 80 °C for 24 hours. On cooling to room temperature, 30 ml of water and 50 ml of DCM were added and the mixture was basified with 1 N NaOH. The layers were separated and the aqueous phase was extracted with 3x10 ml of DCM. The combined organic extracts were treated with 30 ml of 0.1 N HCI solution and the layers separated. The aqueous phase was extracted with 30 ml of DCM and the combined organic phases were evaporated. The resulting residue was purified by chromatography using a Biotage SP1 purification device eluting with EtOAc/cyclohexane and 10 g normal phase silica SNAP coloumn (gradient 25-67% of EtOAc in 20 CV) to give the title compound (5.9 mg); m/z (ES): 441.4 [M+H]+; 1H NMR (500 MHz, CHLOROFORM-d) δ ppm 2.78 (s, 3 H) 5.07 (s, 2 H) 7.53 (dd, J=5.65, 1.83 Hz, 1 H) 7.70 (d, J=5.49 Hz, 1 H) 7.77 (d, J=1.68 Hz, 1 H) 7.94 (d, J=7.02 Hz, 1 H) 8.32 (d, J=5.65 Hz, 1 H) 9.10 (s, 1 H). Compound 31 : N-(2-Chloro-4-Dyridinvn-7-fluoro-4H-n,2,3ltriazolor5,1 - ciri,41benzothiazine-3-carboxamide
Figure imgf000084_0002
To a solution of 4-amino-2-chloropyridine (147 mg, 1.146 mmol) in dry 1 ,4-dioxane (3.00 mL), stirred at room temperature under argon atmosphere, trimethylaluminium, 2 M solution in heptanes (0.573 mL, 1.146 mmol) was added dropwise and the reaction mixture was stirred for 1 hour. Ethyl 7-fluoro-4H-[1 ,2,3]triazolo[5,1- c][1 ,4]benzothiazine-3-carboxylate (intermediate 63, 80 mg, 0.286 mmol) in dry 1 ,4- dioxane (3 mL) was added dropwise and the mixture was stirred at 95 °C overnight. The reaction mixture was cooled to room temperature, quenched with 20 mL of water and diluted with DCM and MeOH. The mixture was evaporated and the resulting residue was partioned between DCM and water. The layers were separated and the aqueous layer was washed with DCM (2x). The combined organic layers were dried using a phase separator filter tube and concentrated under reduced pressure. The resulting residue was purified using a Biotage SP-1 system using a 50 g Si SNAP column eluting under isocratic conditions: 30 % ethyl-acetate / 70 % cyclohexane / 20 CV) to give the title compound (85 mg); 1H NMR (500 MHz, DMSO-c/6): δ ppm 4.69 (s, 2 H) 7.34 (td, J=8.62, 2.75 Hz, 1 H) 7.64 (dd, J=8.85, 2.75 Hz, 1 H) 7.89 (dd, J=5.65, 1.83 Hz, 1 H) 8.06 (d, J=1.68 Hz, 1 H) 8.20 (dd, J=8.93, 5.1 1 Hz, 1 H) 8.33 (d, J=5.65 Hz, 1 H) 1 1 .34 (br. s., 1 H); MS (m/z): 362 [MH]+
Compound 32: 7-Fluoro-N-(2-methyl-4-pyridinyl)-4H-ri ,2,3ltriazolor5,1- cl[1 ,4lbenzothiazine-3-carboxamide
Figure imgf000085_0001
To a solution of 4-amino-2-methylpyridine (124 mg, 1.146 mmol) in dry 1 ,4-dioxane (3 mL) stirred at room temperature under argon was added trimethylaluminium (2M in heptanes) (0.573 mL, 1.146 mmol) and the reaction mixture was stirred for 1 hour. Ethyl 7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3-carboxylate (intermediate 63, 80 mg, 0.286 mmol) in dry 1 ,4-dioxane (3 mL) was added dropwise and the mixture was stirred at 95 °C for 2 days. The reaction mixture was cooled to room temperature, quenched with 20 mL of water and diluted with DCM and MeOH. The mixture was evaporated and the residue was extracted with DCM and water. The layers were separated and the aqueous layer was extracted with with DCM (2x). The combined organic layers were washed with saturated aqueous sodium bicarbonate (2x), dried using a phase separator filter tube and concentrated under reduced pressure. The residue was purified using a Biotage SP-1 system using a 25 g Si SNAP column, eluting with DCM / 2N NH3 in MeOH gradient (0-5 % of 2N NH3 in MeOH / 20 CV) to give a product which was dissolved in 2 mL of EtOAc. A precipitate formed which was collected by filtration and washed twice with diisopropyl ether to give the title compound (30 mg); 1H NMR (500 MHz, DMSO-c/6): δ ppm 2.44 (s, 3 H) 4.69 (s, 2 H) 7.34 (td, J=8.62, 2.90 Hz, 1 H) 7.64 (dd, J=8.85, 2.75 Hz, 1 H) 7.68 (dd, J=5.65, 1 .83 Hz, 1 H) 7.79 (d, J=1.83 Hz, 1 H) 8.19 (dd, J=9.00, 5.19 Hz, 1 H) 8.35 (d, J=5.65 Hz, 1 H) 10.98 (s, 1 H); MS (m/z): 342 [MH]+
Compound 33: N-(2-Chloro-4-pyridinylWluoro-4H-M .2,3ltriazolor5,1 - clH ,41benzothiazine-3-carboxamide 5,5-dioxide
Figure imgf000086_0001
To a solution of 4-amino-2-chloropyridine (127 mg, 0.989 mmol) in dry 1 ,4-dioxane (2.5 mL) stirred at room temperature under argon was added trimethylaluminium (2 M in heptanes) (0.495 mL, 0.989 mmol) dropwise and the reaction mixture was stirred for 1 hour. Ethyl 7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzothiazine-3- carboxylate 5,5-dioxide (intermediate 64, 77 mg, 0.247 mmol) in dry 1 ,4-dioxane (2.5 mL) was added dropwise and the mixture was stirred at 80 °C for 4 hours. On cooling, the reaction mixture was quenched with water and extracted with DCM. 1 N NaOH was added and the layers were separated. The organic phase was washed with 0.1 M HCI and dried using a phase separator filter tube. Evaporation gave a residue (Residue B) which contained mostly starting material together with some title compound. The reaction was repeated to give the title compound (35 mg); 1H NMR (500 MHz, DMSO-d6): δ ppm 5.62 (s, 2 H) 7.88 (dd, J=5.65, 1.68 Hz, 1 H) 7.93 (td, J=8.62, 2.90 Hz, 1 H) 8.06 (d, J=1.68 Hz, 1 H) 8.1 1 (dd, J=7.17, 2.75 Hz, 1 H) 8.36 (d, J=5.65 Hz, 1 H) 8.51 (dd, J=9.00, 4.27 Hz, 1 H) 1 1.50 (s, 1 H); MS (m/z): 394 [MH]+.
Compound 34: N-(2-Chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazor5,1- ciri ,41benzothiazine-3-carboxamide
Figure imgf000086_0002
To a solution of 4-amino-2-chloropyridine (61.6 mg, 0.479 mmol) in dry 1 ,4-dioxane (1.25 mL) at room temperature under argon was added trimethylaluminium (2 M in heptanes) (0.239 mL, 0.479 mmol) dropwise and the reaction mixture was stirred for 1 hour. Ethyl 7-fluoro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 65, 35 mg, 0.120 mmol) in dry 1 ,4-dioxane (1 .25 mL) was added dropwise and the mixture was stirred at 100 °C for 6 hours. On cooling to room temperature, the reaction mixture was quenched with water and extracted with DCM. A 1 M solution of NaOH was added in order to separate the phases. The combined organic layers were washed with 1 N HCI, dried through a phase separator filter tube and concentrated under reduced pressure. The residue contained a significant amount of starting material. The reaction was repeated, this time heating the mixture at 100 degC for 2 days, to give the title compound (6mg) after washing with diethyl ether. Concentrating the ethereal mother liquor gave a further crop of the title compound (4 mg); 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 2.66 (s, 3 H) 4.39 (s, 2 H) 7.08 (ddd, J=8.93, 7.71 , 2.90 Hz, 1 H) 7.30 (dd, J=8.09, 2.90 Hz, 1 H) 7.48 - 7.53 (m, 2 H) 7.80 (d, J=1.83 Hz, 1 H) 8.29 (d, J=5.65 Hz, 1 H) 9.16 (s, 1 H); MS (m/z): 375 [MH]+
Compound 34: altermative synthesis: N-(2-Chloro-4-pyridinyl)-7-fluoro-1-methyl-4H- imidazo[5,1-cl[1 ,41benzothiazine-3-carboxamide
Figure imgf000087_0001
To a solution of 4-amino-2-chloropyridine (54.3 mg, 0.423 mmol) in dry 1 ,4-dioxane (1 mL) stirred at room temperature under argon was added trimethylaluminium (2M in heptanes) (0.21 1 mL, 0.423 mmol) was added dropwise and the reaction mixture was stirred for 1 hour. (7-Fluoro-1-methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazin-3- yl)carbonyl 2-methylpropyl carbonate (intermediate 65, 22 mg, 0.060 mmol) in dry 1 ,4-dioxane (1.000 mL) was added dropwise and the mixture was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature, quenched with water and extracted with DCM. 1 M NaOH was added in order to separate phases. The combined organic layers were washed with 0.1 N HCI, dried through a phase separator filter tube and concentrated under reduced pressure to give the title product (1 1 mg); 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 2.66 (s, 3 H) 4.39 (s, 2 H) 7.08 (ddd, J=8.93, 7.71 , 2.90 Hz, 1 H) 7.30 (dd, J=8.09, 2.75 Hz, 1 H) 7.48 - 7.53 (m, 2 H) 7.80 (d, J=1.68 Hz, 1 H) 8.29 (d, J=5.65 Hz, 1 H) 9.16 (s, 1 H); MS (m/z): 375 [MH]+
Compound 35: N-(2-Chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazor5,1- clH ,41benzothiazine-3-carboxamide 5 5-dioxide
Figure imgf000088_0001
To a solution of N-(2-chloro-4-pyridinyl)-7-fluoro-1-methyl-4H-imidazo[5,1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 34, 14 mg, 0.037 mmol) in a mixture of dry dichloromethane (1 .5 mL) and dry methanol (0.5 mL) was added mCPBA (19.34 mg, 0.1 12 mmol) and the reaction mixture was stirred at room temperature overnight. Further mCPBA (6.5 mg, 0.037 mmol) was added and the mixture was left to stir at room temperature for 1 day. The solvent was evaporated and the residue was dissolved in DCM and washed with saturated aqueous sodium bicarbonate. The organic layer was dried using a phase separator filter tube and concentrated under reduced pressure. The residue was purified by preparative LC/MS followed by passing through a 200 mg SAX column (PS-HCO3) and lyophilized to give the title compound (2.2 mg); 1H NMR (500 MHz, DMSO-c/6): δ ppm 2.78 (s, 3 H) 5.37 (s, 2 H) 7.82 - 8.33 (m, 6 H) 10.87 (br. s., 1 H); MS (m/z): 407 [MH]+.
Compound 36: 7-Fluoro-1 -methyl-N-(2-methyl-4-pyridinyl)-4H-imidazor5,1 - ciri ,41benzothiazine-3-carboxamide
Figure imgf000088_0002
To a solution of 4-amino-2-methylpyridine (74.8 mg, 0.692 mmol) in dry 1 ,4-dioxane (1 mL) stirred at room temperature under argon was added trimethylaluminium (2 M in heptanes) (0.346 mL, 0.692 mmol) dropwise and the reaction mixture was stirred for 1 hour. (7-Fluoro-1-methyl-4H-imidazo[5,1-c][1 ,4]benzothiazin-3-yl)carbonyl 2- methylpropyl carbonate (intermediate 67, 36 mg, 0.099 mmol) in dry 1 ,4-dioxane (1 mL) was added dropwise and the mixture was stirred at 80 °C for 4 hours. The reaction mixture was cooled to room temperature, quenched with water and extracted with DCM. 1 M NaOH was added in order to separate phases. The organic layers were washed with 0.1 M HCI, then dried using a phase separator filter tube and concentrated under reduced pressure to give a residue which contained the title compound with some impurities. The retained aqueous phase was basified with 1 M NaOH and extracted with DCM. The organic layer was dried using a phase separator filter tube and concentrated under reduced pressure to give a residue containing the title compound together with some impurities. This residue was dissolved in DCM, 1 M HCI was added and mixture stirred vigorously for 1 hour. The layers were separated and the water layer basified with 1 M NaOH. The mixture was was extracted with DCM and the organic layer dried using a phase separator filter tube and concentrated under reduced pressure to give the title compound of >90% purity (by NMR). This 90% pure material was further purified. The residue was dissolved in DCM, water was added and mixture stirred vigorously for 1 hour. The organic layer was dried using a phase separator filter tube and concentrated under reduced pressure to give the title compound after washing with diethyl ether; 1H NMR (500 MHz, CHLOROFORM-d): δ ppm 2.56 (s, 3 H) 2.66 (s, 3 H) 4.41 (s, 2 H) 7.05 - 7.10 (m, 1 H) 7.30 (dd, J=8.16, 2.82 Hz, 1 H) 7.41 (dd, J=5.57, 1.91 Hz, 1 H) 7.50 (dd, J=8.93, 4.81 Hz, 1 H) 7.56 (d, J=1 .68 Hz, 1 H) 8.41 (d, J=5.65 Hz, 1 H) 9.04 (s, 1 H); MS (m/z): 355 [MH]+. Compound 37: 6-Chloro-N-(2-chloro-4-Dyridinvn-7-fluoro-4H-n,2,3ltriazolor5,1 - ciri,41benzothiazine-3-carboxamide
Figure imgf000089_0001
To a solution of 4-amino-2-chloropyridine (57.4 mg, 0.446 mmol) in dry 1 ,4-dioxane (2 mL) stirred at room temperature under argon was added trimethylaluminium (2 M in heptanes) (0.223 mL, 0.446 mmol) dropwise and the reaction mixture was stirred for 1 hour. Ethyl 6-chloro-7-fluoro-4H-[1 ,2,3]triazolo[5,1-c][1 ,4]benzothiazine-3- carboxylate (35 mg, 0.1 12 mmol) in dry 1 ,4-dioxane (2.000 mL) was added dropwise and the mixture was stirred at 90 °C for 5 h. The reaction mixture was cooled to room temperature, quenched with water and extracted with DCM. 1 M NaOH was added in order to separate phases. The organic layers were washed with 0.1 M HCI, then dried using a phase separator filter tube and concentrated under reduced pressure. The resulting residue contained significant starting material and so the reaction was repeated by heating the reaction first at 90 °C overnight and then at 100 °C for additional 6 hours and afterwards at 95 °C overnight, to give the title compound (8 mg); 1H NMR (500 MHz, DMSO-d6): δ ppm 4.78 (s, 2 H) 7.56 (t, J=8.93 Hz, 1 H) 7.89 (dd, J=5.65, 1 .68 Hz, 1 H) 8.06 (d, J=1.53 Hz, 1 H) 8.21 (dd, J=9.00, 4.73 Hz, 1 H) 8.33 (d, J=5.65 Hz, 1 H) 1 1 .36 (s, 1 H); MS (m/z): 396 [MH]+
Compound 38: 6-Chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-1 -methyl-4H-imidazo[5,1- cl[1 ,4lbenzothiazine-3-carboxamide
Figure imgf000090_0001
To a solution of 4-amino-2-chloropyridine (78 mg, 0.588 mmol) in dry 1 ,4-dioxane (1.5 mL) stirred at room temperature under argon atmosphere was added
trimethylaluminium (2 M in heptanes) (0.294 mL, 0.588 mmol) dropwise and the reaction mixture was stirred for 1 hour. Ethyl 6-chloro-7-fluoro-1 -methyl-4H- imidazo[5,1-c][1 ,4]benzothiazine-3-carboxylate (intermediate 72, 48 mg, 0.147 mmol) in dry 1 ,4-dioxane (1.5 mL) was added dropwise and the mixture was stirred at 90 °C for 3 h. The reaction mixture was cooled to room temperature, quenched with water and extracted with DCM. 1 M NaOH was added in order to separate phases. The layers were separated and aqueous layer was extracted with DCM (x2). The combined organic layers were washed with 0.1 M HCI, then dried using a phase separator filter tube and concentrated under reduced pressure. The resulting residue was triturated with diethyl ether to give the title compound as a precipitate (43 mg); 1H NMR (500 MHz, DMSO-d6): δ ppm 2.65 (s, 3 H) 4.53 (s, 2 H) 7.52 (t, J=8.85 Hz, 1 H) 7.86 - 7.91 (m, 2 H) 8.07 (d, J=1.53 Hz, 1 H) 8.27 (d, J=5.65 Hz, 1 H) 10.69 (s, 1 H); MS (m/z): 409 [MH]+
The following compounds were also prepared using methods similar to those described herein.
Figure imgf000091_0001
Figure imgf000092_0001
The compounds of the invention may be tested in the following mGluR5 aequorin assay.
Cell Preparation
CHO cells containing human mGluR5 receptors with Tet On expression control technology (supplied by Clontech) were prepared. These cells were grown in cell factories, induced with 10ng/ml doxycycline to enable expression, harvested and then cryo-preserved at -140°C in 1 ml aliquots for future use. On the afternoon prior to the assay, the cells were thawed, suspended in growth media and centrifuged at l OOOrpm for 5 min. The growth media consisted of F12 Hams Nutrient mix (supplied by Gibco - catalogue number 21765) and 10% Tet approved FBS (supplied by Clontech - catalogue number 631 106). The cells were then re-suspended in growth media and incubated at 37°C for 1 hour in a spinner flask. After this post thaw recovery period, the cell suspension was centrifuged once more and resuspended at 2.5 x 10^ cells/ml in loading buffer consisting of HBSS, 0.1 % BSA (supplied by CalBiochem - catalogue number 126609) and 0.1 % Pluronic F68 (supplied by Gibco - catalogue number 24040-032). The cells were loaded with coelentrazine (supplied by Invitrogen C - catalogue number 6780) to a concentration of 5μΜ, wrapped in foil and loaded overnight with mixing. Immediately prior to the assay, the cells were diluted to 15 x 10^ cells/ml in dilution buffer consisting of HBSS and 0.1 % Pluronic F68.
Coelentrazine is the chromophore co-factor which activates the apo-protein, aequorin. The protein has three high affinity binding sites for calcium. Upon agonism of the mGluR5 receptor, binding of calcium to the aequorin protein induces a conformational change resulting in an oxidative decarboxylation reaction producing coelenteramide and a flash luminescence signal. This signal was measured using the Lumilux (supplied by Perkin Elmer).
Compound Preparation
The test compounds were prepared in DMSO at a concentration of 3mM. These solutions are serially diluted with DMSO to 1 in 4 using a Biomek FX liquid handling device (supplied by Beckman Coulter) in a 384-well compound plate (supplied by Greiner). Daughter plates of 0.5μΙΛ/νβΙΙ were stamped-out from this master plate for use in the assay. Glutamate Dose Response Curve Preparation
A 100mM solution of glutamic acid was prepared in water. This was further diluted with DMSO to a concentration of 10mM. 16 x 1 1 point concentration response curves (CRC) were prepared in DMSO, making the final assay concentration 1.66 x10"^M, with 1 in 3 serial dilutions using the Biomek FX. 0.5μΙ stamp-outs of this plate were generated for use in the assay.
Estimation of EC80 concentration of Glutamate
The glutamate CRC plate was placed on the Lumilux where 20μΙΛ/νβΙΙ of dilution buffer was added, followed by 10μΙ/well of loaded cell suspension and a
luminescence read was made. An EC80 concentration of glutamate was calculated by using 4X EC50 generated. The EC80 solution was prepared in dilution buffer and added to a reservoir within the Lumilux.
Running the Assay
20μΙΛ/νβΙΙ of buffer and 10μΙ/well of cell suspension were added to the test compounds using the lumilux. These plates were incubated at room temperature for 15 mins, 10μΙ/well of EC80 solution added and a luminescence read made. Blocking of the receptor by test compound in a dose dependent manner was evident from the luminescence vs time profiles generated for each well. The data was analysed using XC50 software to produce CRCs, from which the potency and plC50 can be determined. When tested in this assay, supporting compounds 1 -20, 22-39, 42-54 gave a plC50 of 6.5 to 9.1. Supporting compound 37 gave a plC50 of 9.1. Supporting Compounds 55 and 56 were not tested in this assay.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof,
Figure imgf000095_0001
(I)
wherein
E is -CH2-, -0-, -S-, -(S=0)- or -SO2-;
ring A is either:
a) a triazole ring where V is C, W is C, X is N, Y is N and Z is N;
b) an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is
N; or
c) an oxazole ring where V is C, W is C, X is C, Y is O and Z is N; wherein when ring A is imidazole, RY is H, C-|.galkyl, C-|.ghaloalkyl,
C3_gcycloalkyl or C3_gcycloalkyl(C-| _4)alkyl;
n is 1 or 2;
R1 is phenyl or 5- or 6-membered monocyclic heteroaryl, either of which are optionally substituted by one or two groups independently selected from halo, C-|.galkyl, C-|.galkoxy, C-|.ghaloalkyl, C-|.ghaloalkoxy, cyano, C-|.galkylthio, C-|.ghaloalkylthio, C-|.galkylsulfonyl, C-|.ghaloalkylsulfonyl, C-|.galkylcarbonyl and C-|.ghaloalkylcarbonyl; and
R2, R3, R4 and R5, which may be the same or different, are selected from the list consisting of H, halo, C-|.galkyl, C-|.galkoxy, C-|.ghaloalkyl, C-|. ghaloalkoxy, cyano, C-|.galkylthio and C-|.ghaloalkylthio, wherein two or more of R2, R3, R4 and R5 are H.
2. A compound according to claim 1 or a pharmaceutically acceptable salt
thereof, wherein E is -CH2-, -0-, -S- or -SC>2-- 3. A compound according to any preceding claim or a pharmaceutically
acceptable salt thereof wherein ring A is: a) a triazole ring where V is C, W is C, X is N, Y is N and Z is N; or b) an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; wherein when ring A is imidazole, RY is C-|.galkyl, C-|.ghaloalkyl or
C3_gcycloalkyl.
A compound according to claim 1 or claim 2 or a pharmaceutically acceptable salt thereof wherein ring A is a triazole ring where V is C, W is C, X is N, Y is N and Z is N.
A compound according to claim 1 or claim 2 or a pharmaceutically acceptable salt thereof, wherein ring A is an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; wherein RY is C-|.galkyl, C-|.ghaloalkyl or
C3_gcycloalkyl.
A compound according to any preceding claim or a pharmaceutically acceptable salt thereof, wherein R1 is pyridyl optionally substituted by halo or alkyl.
A compound according to any preceding claim or a pharmaceutically acceptable salt thereof, wherein R2, R3j R4 and R5j which may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R2, R3, R4 and R5 are H.
A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein
E is -CH2-, -0-, -S- or -SO2-;
ring A is a triazole ring where V is C, W is C, X is N, Y is N and Z is N;
n is 1 or 2;
R1 is pyridyl optionally substituted by halo or alkyl; and
R2, R3, R4 and R5, which may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R2, R^, R4 and
R5 are H.
A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein
E is -CH2-, -0-, -S- or -SO2-; ring A is an imidazole ring where V is C, W is C, X is N, Y is -C(RY)- and Z is N; wherein RY is C-|.galkyl, C-|.ghaloalkyl or C3_gcycloalkyl; n is 1 or 2;
R1 is pyridyl optionally substituted by halo or alkyl; and
R2, R3, R4 ANC| R5J which may be the same or different, are selected from the list consisting of H and halo, wherein two or more of R^, R3, R4 ANC|
R5 are H.
A compound according to claim 1 selected from the list:
8-chloro-N-(2-methyl-4-pyridinyl)-4H-[1 ,2,3]triazolo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 1 );
8-chloro-N-(2-chloro-4-pyridinyl)-4H-[1 ,2,
3]triazolo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 2);
8-chloro-1-methyl-/V-(2-methyl-4-pyridinyl)-4/-/-imidazo[5, 1 - c][1 ,4]benzothiazine-3-carboxamide 5,5-dioxide (Compound 3);
8-chloro-1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5, 1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 4);
8-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 5);
8-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1 - c][1 ,
4]benzothiazine-3-carboxamide 5,
5-dioxide
(Compound 6);
8-chloro-/V-(2-chloro-4-pyridinyl)-7-f luoro-1 -methyl-4/-/-imidazo[5, 1 - c][1 ,4]benzoxazine-3-carboxamide
(Compound 7);
8-chloro-/V-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 ,2,3]triazolo[5, 1 - c][1 ,4]benzoxazine-3-carboxamide (Compound 8);
N-(2-chloro-4-pyridinyl)-5,6-dihydro-4H-[1 ,2,3]triazolo[1 ,5-a][1]benzazepine-3- carboxamide
(Compound 9);
N-(2-chloro-4-pyridinyl)-1 -methyl-5,6-dihydro-4H-imidazo[1 ,5- a][1]benzazepine-3-carboxamide
(Compound 10);
N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 1 1 );
1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide (Compound 12);
N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5, 1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 13); N-(6-chloro-2-pyridinyl)-1 -methyl-4H-imidazo[5,1 -c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 14);
N-(2-chloro-4-pyridinyl)-4H-[1 !2!3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide (Compound 15);
N-(2-chloro-4-pyridinyl)-4H-[1 !2,3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 16);
N-(2-methyl-4-pyridinyl)-4H-[1 !2!3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide (Compound 17);
N-(2-chloro-4-pyridinyl)-1 -(trifluoromethyl)-4H-imidazo[5,1- c][1 ,4]benzothiazine-3-carboxamide (Compound 18);
N-(2-chloro-4-pyridinyl)-1 -cyclopropyl-4H-imidazo[5,1-c][1 ,4]benzothiazine-3- carboxamide (Compound 19);
N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzoxazine-3- carboxamide (Compound 20);
N-(6-chloro-2-pyridinyl)-7-fluoro-4H-[1 ,2,3]triazolo[5,1 -c][1 ,4]benzoxazine-3- carboxamide (Compound 21 );
N-(2-chloro-4-pyridinyl)-7-fluoro-1 -methyl-4H-imidazo[5,1-c][1 ,4]benzoxazine-
3-carboxamide (Compound 22);
7-chloro-N-(2-chloro-4-pyridinyl)-4H-[1 !2!3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide (Compound 23);
7-chloro-N-(2-methyl-4-pyridinyl)-4H-[1 !2,3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide (Compound 24);
7-chloro-N-(2-chloro-4-pyridinyl)-1-methyl-4H-imidazo[5,1- c][1 ,4]benzothiazine-3-carboxamide (Compound 25);
7-chloro-N-(2-chloro-4-pyridinyl)-4H-[1 !2!3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 26);
7- chloro-1-methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5,1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 27);
8- chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 !2!3]triazolo[5,1- c][1 ,4]benzothiazine-3-carboxamide (Compound 28);
8-chloro-N-(2-chloro-4-pyridinyl)-7-f luoro-1 -methyl-4H-imidazo[5, 1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 29);
8-chloro-N-(2-chloro-4-pyridinyl)-7-f luoro-1 -methyl-4H-imidazo[5, 1 - c][1 ,4]benzothiazine-3-carboxamide 5,5-dioxide (Compound 30); N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 !2!3]triazolo[5!1 -c][1 ,4]benzothiazine-3- carboxamide (Compound 31 ); 7-fluoro-N-(2-methyl-4-pyridinyl)-4H-[1 !2!3]triazolo[5!1-c][1 ,4]benzothiazine-3- carboxamide (Compound 32);
N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 !2!3]triazolo[5!1 -c][1 ,4]benzothiazine-3- carboxamide 5,5-dioxide (Compound 33);
N-(2-chloro-4-pyridinyl)-7-fluoro-1 -methyl-4H-imidazo[5,1- c][1 ,4]benzothiazine-3-carboxamide (Compound 34);
N-(2-chloro-4-pyridinyl)-7-fluoro-1 -methyl-4H-imidazo[5,1- c][1 ,4]benzothiazine-3-carboxamide 5,5-dioxide (Compound 35); 7-fluoro-1 -methyl-N-(2-methyl-4-pyridinyl)-4H-imidazo[5,1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 36);
6-chloro-N-(2-chloro-4-pyridinyl)-7-fluoro-4H-[1 !2!3]triazolo[5,1- c][1 ,4]benzothiazine-3-carboxamide (Compound 37); and 6-chloro-N-(2-chloro-4-pyridinyl)-7-f luoro-1 -methyl-4H-imidazo[5, 1 - c][1 ,4]benzothiazine-3-carboxamide (Compound 38);
or a pharmaceutically acceptable salt thereof.
1 1 . A compound as defined in any preceding claim or a salt thereof for use in therapy.
12. A compound as defined in any one of claims 1 to 10 or a salt thereof for use in treating a substance related disorder.
13. A use of a compound defined in any one of claims 1 to 10 or a salt thereof, in the manufacture of a medicament for treating a substance related disorder.
14. A method of treating a substance related disorder in a mammal (preferably a human) comprising administering an effective amount of a compound defined in any one of claims 1 to 10 or a salt thereof.
15. A composition comprising a) a compound defined in any one of claims 1 to 10 or a salt thereof, and b) a pharmaceutically acceptable excipient.
PCT/EP2011/059011 2010-06-03 2011-06-01 Novel compounds WO2011151361A1 (en)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
GB1009341.7 2010-06-03
GBGB1009341.7A GB201009341D0 (en) 2010-06-03 2010-06-03 Novel compounds
GBGB1019575.8A GB201019575D0 (en) 2010-11-18 2010-11-18 Novel compounds
GB1019575.8 2010-11-18
GB1020622.5 2010-12-06
GBGB1020622.5A GB201020622D0 (en) 2010-12-06 2010-12-06 Novel compounds

Publications (1)

Publication Number Publication Date
WO2011151361A1 true WO2011151361A1 (en) 2011-12-08

Family

ID=44227962

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2011/059011 WO2011151361A1 (en) 2010-06-03 2011-06-01 Novel compounds

Country Status (1)

Country Link
WO (1) WO2011151361A1 (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014209978A1 (en) * 2013-06-24 2014-12-31 Merck Patent Gmbh Imidazole compounds as modulators of fshr and uses thereof
CN104788474A (en) * 2015-03-28 2015-07-22 陈定奔 Synthesis method of triazole benzothiazinone fused heterocycle
WO2015177646A1 (en) 2014-05-14 2015-11-26 Novartis Ag Carboxamide derivatives
US9403810B2 (en) 2014-05-14 2016-08-02 Novartis Ag Carboxamide derivatives
CN111454229A (en) * 2020-04-10 2020-07-28 颜建发 Dihydronaphthoisoxazole derivative and application thereof in antitumor drugs
CN113045557A (en) * 2021-03-08 2021-06-29 宁夏一帆生物科技有限公司 Production process of flumioxazin herbicide

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0827131A (en) * 1994-07-11 1996-01-30 Teikoku Hormone Mfg Co Ltd New bicyclic compound-condensed (2,1-d)isoxazole-3-carboxamide derivative
WO2010049366A1 (en) * 2008-10-27 2010-05-06 Glaxo Group Limited Tricyclic compounds as glutamate receptor modulators

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0827131A (en) * 1994-07-11 1996-01-30 Teikoku Hormone Mfg Co Ltd New bicyclic compound-condensed (2,1-d)isoxazole-3-carboxamide derivative
WO2010049366A1 (en) * 2008-10-27 2010-05-06 Glaxo Group Limited Tricyclic compounds as glutamate receptor modulators

Non-Patent Citations (11)

* Cited by examiner, † Cited by third party
Title
"Diagnostic and Statistical Manual of Mental Disorders", AMERICAN PSYCHIATRIC ASSOCIATION
"International Classification of Diseases"
"Remington's Pharmaceutical Sciences", MACK PUBLISHING COMPANY
"Topics in Chemistry", pages: 306 - 316
BERGE ET AL., J. PHARM, SCI., vol. 66, 1977, pages 1 - 19
BIGHLEY ET AL.: "Encyclopedia of Pharmaceutical Technology", vol. 13, 1996, MARCEL DEKKER INC, pages: 453 - 497
DRUGS OF TODAY, vol. 19, no. 9, 1983, pages 499 - 538
FRAVOLINI ET AL: "Synthesis and pharmacological activity of benzothiopyranopyrazole and benzothiopyranoisoxazole carboxamides", FARMACO, EDIZIONE SCIENTIFICA, SOCIETA CHIMICA ITALIANA, PAVIA, IT, vol. 33, no. 11, 1 January 1978 (1978-01-01), pages 855 - 865, XP002146285, ISSN: 0430-0920 *
H. BUNDGAARD, DESIGN OF PRODRUGS
H. BUNDGAARD: "Design of Prodrugs", 1985, ELSEVIER
P L GOULD, INTERNATIONAL JOURNAL OF PHARMACEUTICS, vol. 33, 1986, pages 201 - 217

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016523864A (en) * 2013-06-24 2016-08-12 メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフツングＭｅｒｃｋ Ｐａｔｅｎｔ Ｇｅｓｅｌｌｓｃｈａｆｔ ｍｉｔ ｂｅｓｃｈｒａｅｎｋｔｅｒ Ｈａｆｔｕｎｇ Imidazole compounds and their use as regulators of FSHR
WO2014209978A1 (en) * 2013-06-24 2014-12-31 Merck Patent Gmbh Imidazole compounds as modulators of fshr and uses thereof
US10138233B2 (en) 2013-06-24 2018-11-27 Merck Patent Gmbh Imidazole compounds as modulators of FSHR and uses thereof
AU2014302710B2 (en) * 2013-06-24 2018-10-04 Merck Patent Gmbh Imidazole compounds as modulators of FSHR and uses thereof
CN106536512B (en) * 2014-05-14 2019-06-07 诺华股份有限公司 Carboxamides derivatives
WO2015177646A1 (en) 2014-05-14 2015-11-26 Novartis Ag Carboxamide derivatives
CN106536512A (en) * 2014-05-14 2017-03-22 诺华股份有限公司 Carboxamide derivatives
CN106661012A (en) * 2014-05-14 2017-05-10 诺华股份有限公司 Carboxamide derivatives
JP2017515845A (en) * 2014-05-14 2017-06-15 ノバルティス アーゲー Carboxamide derivatives
AU2015263049B2 (en) * 2014-05-14 2017-09-28 Novartis Ag Carboxamide derivatives
US11672782B2 (en) 2014-05-14 2023-06-13 Novartis Ag Carboxamide derivatives
US9931319B2 (en) 2014-05-14 2018-04-03 Novartis Ag Carboxamide derivatives
US9403833B2 (en) 2014-05-14 2016-08-02 Novartis Ag Carboxamide derivatives
US9403810B2 (en) 2014-05-14 2016-08-02 Novartis Ag Carboxamide derivatives
US10195181B2 (en) 2014-05-14 2019-02-05 Novartis Ag Carboxamide derivatives
CN106661012B (en) * 2014-05-14 2019-12-20 诺华股份有限公司 Carboxamide derivatives
CN104788474A (en) * 2015-03-28 2015-07-22 陈定奔 Synthesis method of triazole benzothiazinone fused heterocycle
CN104788474B (en) * 2015-03-28 2017-10-10 陈定奔 A kind of synthetic method of triazole benzo buprofezin condensed hetero ring
CN111454229A (en) * 2020-04-10 2020-07-28 颜建发 Dihydronaphthoisoxazole derivative and application thereof in antitumor drugs
CN111454229B (en) * 2020-04-10 2022-07-19 徐瑛 Dihydronaphthoisoxazole derivative and application thereof in antitumor drugs
CN113045557A (en) * 2021-03-08 2021-06-29 宁夏一帆生物科技有限公司 Production process of flumioxazin herbicide

Similar Documents

Publication Publication Date Title
JP5820537B2 (en) Substituted pyrazoles as MGLU5 receptor modulators
IL181387A (en) Fused tricyclic derivatives for the treatment of psychotic disorders
WO2011151361A1 (en) Novel compounds
WO2011012622A1 (en) Benzoxazinone derivatives for the treatment of glytl mediated disorders
US4663453A (en) Benzo[b]pyrrolo[3,2,1-jk][1,4]benzodiazepines having dopamine receptor activity
Hussenether et al. Clozapine derived 2, 3-dihydro-1H-1, 4-and 1, 5-benzodiazepines with D4 receptor selectivity: synthesis and biological testing
WO2009100536A1 (en) Inhibitors of kinase activity with 1,2-di-cyclyl substituted alkyne structures
EP2488514A1 (en) Indole and azaindole modulators of the alpha 7 nachr
CA2569088C (en) Pyrrolobenzimidazolones and their use as antiproliferative agents
JPWO2020145250A1 (en) 15-PGDH inhibitor
WO2009115486A1 (en) Triazole amide derivatives for use in therapy
WO2003057161A2 (en) BENZOTHIENO [3,2-c]PYRAZOLYL AND BENZOFURANO [3,2-c] PYRAZOLYL COMPOUNDS, THEIR USE IN DISEASES ASSOCIATED WITH THE 5-HT2C RECEPTOR AND INTERMEDIATE COMPOUNDS THEREOF
CA2316388C (en) Tetrahydrobenzindole derivative
WO2011056739A1 (en) Compounds and methods
CA2797019C (en) Modulators of 5-ht receptors and methods of use thereof
CA2713551C (en) Triazolium salts as par1 inhibitors, production thereof, and use as medicaments
JPS61286367A (en) Benzazepine derivative
TW200825083A (en) Novel fluorene derivatives, compositions containing them and use thereof
NO873108L (en) PROCEDURE FOR THE PREPARATION OF THERAPEUTIC ACTIVE 1,4-BENZODIAZEPINES.
US8119629B2 (en) Carboxamide GABAA α2 modulators
WO2011023753A1 (en) Benzoxazine derivatives as glycine transport inhibitors
AU2009211887B2 (en) Triazolopyridazines as PAR1 inhibitors, production thereof, and use as medicaments
CA2562561A1 (en) 6-substituted pyridoindolone derivatives production and therapeutic use thereof
WO2010142652A1 (en) Imidazobenzazepine compounds for the treatment of central nervous system (cns) diseases
US4761412A (en) Benzopyrrolobenzodiazepines and quinobenzodiazepines useful for the treatment of psychoses

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11724169

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11724169

Country of ref document: EP

Kind code of ref document: A1