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WO2025006293A2 - Inhibiteurs du virus respiratoire syncytial humain et du métapneumovirus - Google Patents

Inhibiteurs du virus respiratoire syncytial humain et du métapneumovirus Download PDF

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Publication number
WO2025006293A2
WO2025006293A2 PCT/US2024/034649 US2024034649W WO2025006293A2 WO 2025006293 A2 WO2025006293 A2 WO 2025006293A2 US 2024034649 W US2024034649 W US 2024034649W WO 2025006293 A2 WO2025006293 A2 WO 2025006293A2
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mmol
mixture
pyridin
methyl
concentrated
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PCT/US2024/034649
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English (en)
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WO2025006293A3 (fr
Inventor
Douglas C. Beshore
Brett R. AMBLER
Kira A. Armacost
Christopher James BUNGARD
Danielle M. Hurzy
Peter J. Manley
Kelly-Ann S. Schlegel
Linda M. Suen-Lai
Mahdieh YAZDANI
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Merck Sharp & Dohme Llc
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Publication of WO2025006293A2 publication Critical patent/WO2025006293A2/fr
Publication of WO2025006293A3 publication Critical patent/WO2025006293A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the present invention relates to therapeutic compounds useful for the inhibition of respiratory syncytial virus replication and metapneumovirus replication.
  • the therapeutic compounds may be used in the treatment or prevention of respiratory syncytial virus infection and metapneumovirus infection.
  • Paramyxoviruses are enveloped negative-strand RNA viruses that are significant human and animal pathogens.
  • Human Respiratory Syncytial Virus (hRSV, RSV) belongs to the family Paramyxoviridae, subfamily Pneumovirinae. Two subtypes, type A and type B, have been identified and are a major cause of severe and sometimes even fatal respiratory infections in children less than 6 months of age.
  • Adults with underlying diseases, such as COPD, asthma, cancer, immunocompromised status, including HIV or post transplantation, are also at risk of developing severe RSV infections. Fifteen percent of annual hospitalizations in adults over 50 years of age are due to acute respiratory infections caused by RSV.
  • RSV causes more than 100,000 hospitalizations annually and is estimated to cause 160,000 deaths globally each year.
  • Other viral family members including human metapneumovirus (hMPV) and human Parainfluenza Virus (hPIV), are also responsible for acute respiratory illness similar to hRSV.
  • hMPV human metapneumovirus
  • hPIV human Parainfluenza Virus
  • the RSV genome is a single-stranded negative-sense RNA molecule of approximately 15 kb, which encodes for 11 proteins. Two of these proteins are the main surface glycoproteins of the virion.
  • attachment protein which mediates virus binding to cells
  • F fusion protein
  • hMPV is the second most common cause of lower respiratory infection in young children.
  • hMPV is responsible for 5-40% of respiratory tract infections in hospitalized and outpatient children.
  • hMPV generally results in mild respiratory tract infections;
  • adults 70 years old and older, immunocompromised individuals and, people with comorbidities such as asthma and chronic obstructive pulmonary disease (COPD) are at higher risk for more serious disease and hospitalization as a result of hMPV infection.
  • COPD chronic obstructive pulmonary disease
  • the hMPV genome is approximately 13 kb and the organization is similar to hRSV.
  • hMPV genome RNA replication and mRNA transcription relies on the hMPV L-protein polymerase, which is highly homologous with the hRSV L-protein polymerase.
  • the monoclonal antibody palivizumab (Synagis®) is approved for prophylactic use but has limited efficacy and its use is limited to high-risk infants as a result of high cost.
  • Ribavarin (ViraZole®), a guanosine nucleoside analog broad-spectrum antiviral is approved as an inhaled treatment for RSV infection in infants, but clear efficacy data is lacking (Fearns et al., 2016 Antiviral Research, 134:63-76). In addition, the teratogenic potential of ribavirin raises significant risks for caregivers. The standard of care currently for RSV-infected patients is palliative and includes supplemental oxygen and intravenous fluids. [0007] There continues to be a need for anti-viral agents with pharmacokinetic properties suitable for a significant number of patients in the affected population.
  • the present invention provides novel replication inhibitors of hRSV and hMPV useful for the inhibition of respiratory syncytial virus replication and metapneumovirus replication for addressing this need.
  • SUMMARY OF THE INVENTION [0008]
  • the present disclosure is directed to compounds of Formula I and embodiments thereof for use as anti-viral agents for inhibition of the replication of hRSV and hMPV and the treatment and prophylaxis of hRSV and hMPV infection.
  • Compositions and methods of use comprising the compounds of this disclosure are also provided.
  • the present disclosure is directed to compounds of Formula I (I) i s selected from -H, halo, -NH2, -N(CH3)2, -C 1-3 alkyl, -OC 1-3 alkyl, and -C 3-5 cycloalkyl, wherein any alkyl or cycloalkyl moiety is unsubstituted or substituted with 1 to 3 substituents independently selected at each occurrence from -OH, -OC 1-3 alkyl, and 1, 2 or 3 of fluoro;
  • X 1 is independently selected from C and N;
  • X 2 is independently selected from -CH, O, S, and N;
  • X 3 is independently selected from C and N;
  • X 4 and X 5 are each independently selected from -CH and N;
  • X 6 is independently selected from -CH and N;
  • R 2 is selected from (a) -H, (b) halo, ( c) -C 1-4 alkyl, (d)
  • R 1 is selected from -H, halo, -N(CH3)2, cyclopentane, and a C 1-2 alkyl unsubstituted or substituted with 1 to 2 substituents independently selected from -OH, -OC 1-3 alkyl, and 1, 2 or 3 of fluoro;
  • X 1 and X 3 are C;
  • X 2 is independently selected from -CH, O, S, and N;
  • X 4 is N;
  • X 5 and X 6 are -CH;
  • R 2 is selected from (a) -H, (b) halo, ( c) -C 1-4 alkyl, (d) -OH, -OC 1-3 alkyl, or -OC 3 cycloalkyl, and (e) a 5-member aromatic heterocyclyl ring comprised of three carbon atoms and N and one of O, or NH and O, wherein any R 2
  • Embodiment 1 of this disclosure are compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein: (i) X 1 and X 3 are C; (ii) X 2 is independently selected from -CH, O, S, and N; (iii) X 4 is N; or (iv) X 5 and X 6 are -CH.
  • compounds of Formula I, or a pharmaceutically acceptable salt thereof wherein: (i) X 1 and X 3 are C; (ii) X 2 is independently selected from -CH, O, S, and N; (iii) X 4 is N; and (iv) X 5 and X 6 are -CH.
  • a first sub-class of compounds of Formula I and Embodiment 1 are compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein: X 1 and X 2 are -CH, X 3 is C, X 4 is N, and X 6 is -CH.
  • a first sub-class of compounds of Formula I and Embodiment 1 are compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein: X 1 and X 2 are -CH, X 3 is N, X 4 is -CH, and X 6 is -CH.
  • Embodiment 2 of this disclosure are compounds of Formula I and Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from -H, halo, -N(CH3)2, cyclopentane, and a C 1-2 alkyl unsubstituted or substituted with 1 to 2 substituents independently selected from -OH, -OC 1-3 alkyl, and 1, 2 or 3 of fluoro.
  • R1 is a C 1-2 alkyl unsubstituted or substituted with 1 to 2 substituents independently selected at each occurrence from -OH, methoxy, and 1, 2 or 3 of fluoro.
  • R 1 is selected from -H, -F, -CF 3 , cyclopentane, and C 1-3 alkyl.
  • R 2 is selected from: (a) -H, (b) halo, (c) -C1-4alkyl, ( d) -OH, -OC 1-3 alkyl, or -OC 3 cycloalkyl, and (e) a 5-member aromatic heterocyclyl ring comprised of three carbon atoms and N and one of O, or NH and O, wherein any R 2 alkyl moiety is optionally substituted with 1 to 4 substituents independently selected at occurrence from -OH, methoxy, and 1, 2 or 3 of fluoro.
  • R2 is a C 1-2 alkyl unsubstituted or substituted with 1 to 2 substituents independently selected at each occurrence from -OH, methoxy, and 1, 2 or 3 of fluoro.
  • R2 is a C 1-2 alkyl substituted with -OH.
  • R 2 is selected from -H, -F, -CF3, cyclopentane, and unsubstituted C 1- 3 alkyl.
  • R 1 and R 2 are each C 1-2 alkyl unsubstituted or substituted with 1 to 2 substituents independently selected at each occurrence from -OH, methoxy, and 1, 2 or 3 of fluoro.
  • R 1 and R 2 are each independently selected from -H, -F, -CF 3 , cyclopentane, and C 1-3 alkyl.
  • R 1 and/or R 2 are cyclopentane.
  • R 1 and/or R 2 are -CF3. -In some sub-embodiments, R 1 and/or R 2 are -F.
  • R 2 is selected from: 25700 , X b is independently selected from S, O, and NH, X c is independently selected from N and -CH, X d is independently selected from S and O, X e is independently selected from O and -CH, and R a is selected from -H, methyl, and methoxy.
  • R 2 is selected from -CO2CH3, -OCH3, -OCHF2, -NH 2 , and -N(CH 3 ) 2 .
  • R2 is a C 3-4 cycloalkyl unsubstituted or substituted with substituted with 1 to 4 substituents independently selected at each occurrence from -OH, methoxy, and 1, 2 or 3 of fluoro.
  • Embodiment 4A of this disclosure as well as Embodiments 1-3, and classes thereof, are compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein R 3 is -NH-.
  • Embodiment 4B of this disclosure are compounds of Formula I, and each of Embodiments 1-3, and classes thereof, or a pharmaceutically acceptable salt thereof, wherein R 3 is -O-.
  • Embodiment 5 of this disclosure are compounds of Formula I, and each of Embodiments 1-4A and 4B, and classes thereof, or a pharmaceutically acceptable salt thereof, wherein X 7 is N, and X 8 is -CH.
  • Embodiment 6 of this disclosure are compounds of Formula I, and each of Embodiments 1-5, and classes thereof, or a pharmaceutically acceptable salt thereof, wherein R 4 and R 6 are -H.
  • Embodiment 7 of this disclosure are compounds of Formula I, and each of Embodiments 1-6, and classes thereof, or a pharmaceutically acceptable salt thereof, wherein R 5 25700 is selected from -C 1-3 alkyl unsubstituted or substituted with 1 to 3 substituents independently selected at each occurrence from halo and -NH 2 .
  • R 5 is a C 3 alkyl substituted with -NH2.
  • Embodiment 8 of this disclosure are compounds of Formula I, and each of Embodiments 1-7, and classes thereof, or a pharmaceutically acceptable salt thereof, wherein R 7 is selected from: (1) phenyl substituted with 1 to 3 substituents independently selected at each occurrence from: (a) fluoro, (b) -CN, ( c) -C 1-3 alkyl, or (d) -OC 1-3 alkyl, wherein any such alkyl moiety is optionally substituted with 1 to 3 substituents independently selected at each occurrence from 1, 2 or 3 of fluoro; each selected from -H, -C 1-2 alkyl, methoxy, -CN, and fluoro; w herein R 13a and R 13b are each selected from -H, methyl, -F, and , wherein R 16A and R 16B are each selected from -H or -C1-3 alkyl, substituted with 1, 2 or 3 of fluoro.
  • R 7 is , wherein Rb is selected from -H, halo, -CN, -CF3, and methoxy.
  • Rb is selected from -H, halo, -CN, -CF3, and methoxy.
  • X 1 is N: there is a single bond between X 1 and X 2 and a double bond between X 2 and X 4 .
  • X 2 , X 4 , X 5 and X 6 each 3 -CH, and X is C.
  • a compound is of Formula II: or a [0027] In some aspects of the disclosed compounds, provided herein is a compound is of Formula III: or a as above. [0028] Reference to the compounds of Formula I herein encompasses the compounds of Formula II and III, and all embodiments, classes and sub-classes thereof and includes the compounds of the Examples herein. The compounds of Formula I encompass neutral compounds or salts thereof when such salts are possible, including pharmaceutically acceptable salts.
  • alkyl refers to both branched- and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms in a specified range.
  • C 1-3 alkyl has 1, 2 or 3 carbon atoms and includes each of n-propyl, i-propyl, ethyl and methyl.
  • Cycloalkyl refers to a cyclized alkyl ring having the indicated number of carbon atoms in a specified range.
  • C 3-5 cycloalkyl includes each of cyclopropyl, cyclobutyl, and cyclopentyl
  • C 3-4 cycloalkyl includes each of cyclopropyl and cyclobutyl.
  • Halo or “halogen” refers to chloro, fluoro, bromo or iodo.
  • a “stable” compound is a compound which can be prepared and isolated and whose structure and properties remain or can be caused to remain essentially unchanged for a period of time sufficient to allow use of the compound for the purposes described herein (e.g., therapeutic or prophylactic administration to a subject).
  • the compounds of the present disclosure are limited to stable compounds embraced by Formula I and its embodiments.
  • certain moieties as defined in Formula I may be unsubstituted or substituted, and the latter is intended to encompass substitution patterns (i.e., number and kind of substituents) that are chemically possible for the moiety and that result in a stable compound.
  • This disclosure includes individual diastereomers, particularly epimers, i.e., compounds having the same chemical formula but which differ in the spatial arrangement around a single atom.
  • This disclosure also includes mixtures of diastereomers, particularly mixtures of epimers, in all ratios.
  • This disclosure encompasses compounds of Formula I having either the (R) or (S) stereo-configuration at an asymmetric center and at any additional asymmetric centers that may be present in a compound of Formula I, as well as stereo-isomeric mixtures thereof.
  • Embodiments of this disclosure also include a mixture of enantiomers enriched with 51% or more of one of the enantiomers, including for example 60% or more, 70% or more, 80% or more, or 90% or more of one enantiomer.
  • a single epimer is preferred.
  • An individual or single 25700 enantiomer refers to an enantiomer obtained by chiral synthesis and/or using generally known separation and purification techniques, and which may be 100% of one enantiomer or may contain small amounts (e.g., 10% or less) of the opposite enantiomer.
  • individual enantiomers are a subject of this disclosure in pure form, both as levorotatory and as dextrorotatory antipodes, in the form of racemates and in the form of mixtures of the two enantiomers in all ratios.
  • this disclosure includes both the cis form and the trans form as well as mixtures of these forms in all ratios.
  • the preparation of individual stereoisomers can be carried out, if desired, by separation of a mixture by customary methods, for example by chromatography or crystallization, by the use of stereochemically uniform starting materials for the synthesis or by stereoselective synthesis.
  • a derivatization can be carried out before a separation of stereoisomers.
  • the separation of a mixture of stereoisomers can be carried out at an intermediate step during the synthesis of a compound of Formula I or it can be done on a final racemic product.
  • Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing a stereogenic center of known configuration.
  • absolute stereochemistry may be determined by Vibrational Circular Dichroism (VCD) spectroscopy analysis.
  • VCD Vibrational Circular Dichroism
  • the atoms in a compound of Formula I may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature.
  • the present disclosure is meant to include all suitable isotopic variations of the compounds of Formula I; for example, different isotopic forms of hydrogen (H) include protium ( 1 H) and deuterium ( 2 H). Protium is the predominant hydrogen isotope found in nature. Enriching for deuterium may afford certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound 25700 useful as a standard for characterization of biological samples.
  • Isotopically-enriched compounds of Formula I can be prepared without undue experimentation by conventional techniques well known to those skilled in the art or by processes analogous to those described in the Schemes and Examples herein using appropriate isotopically-enriched reagents and/or intermediates.
  • the compounds can be administered in the form of pharmaceutically acceptable salts.
  • pharmaceutically acceptable salt refers to a salt which possesses the effectiveness of the parent compound and which is not biologically or otherwise undesirable (e.g., is neither toxic nor otherwise rious to the recipient thereof).
  • the invention includes the corresponding pharmaceutically acceptable salts.
  • the compounds of Formula I, and pharmaceutically acceptable salts thereof, which contain one or more basic groups, i.e., groups which can be protonated, can be used according to the invention in the form of their acid addition salts with inorganic or organic acids as, for example but not limited to, salts with hydrogen chloride, hydrogen fluoride, hydrogen bromide, trifluoroacetic acid (trifluoroacetate), phosphoric acid, sulfuric acid, nitric acid, benzenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acids, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfaminic acid, phenylprop
  • one or more NH 2 groups (e.g., a single NH2 group, or two NH2 groups) of any of the compounds of Formula I is protonated in a salt form.
  • one or more NH2 groups (e.g., a single NH2 group) of any of the compounds of Formula I is protonated in a salt form with trifluoroacetic acid (trifluoroacetate).
  • one or more NH2 groups (e.g., a single NH2 group) of any of the compounds of Formula I is protonated in a salt form with hydrogen chloride.
  • the invention also includes, in addition to the salt forms mentioned, inner salts or betaines (zwitterions). Salts can be obtained from the compounds of Formula I by customary methods which are known to the person skilled in the art, for example by combination with an organic or inorganic acid or base in a solvent or dispersant, or by anion exchange or cation exchange from other salts.
  • the present disclosure also includes all salts of the compounds of Formula I which, owing to low physiological compatibility, are not directly suitable for use in pharmaceuticals but which can be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.
  • compositions comprised of a compound of Formula I or a compound that is a salt thereof, including for example but not limited to, a composition comprised of said compound associated together with one or more additional molecular and/or ionic component(s) which may be referred to as a “co-crystal.”
  • co- crystal refers to a solid phase (which may or may not be crystalline) wherein two or more different molecular and/or ionic components (generally in a stoichiometric ratio) are held together by non-ionic interactions including but not limited to hydrogen-bonding, dipole-dipole interactions, dipole-quadrupole interactions or dispersion forces (van der Waals).
  • compounds of the present disclosure may exist in amorphous form and/or one or more crystalline forms, and as such all amorphous and crystalline forms and mixtures thereof of the compounds of Formula I and salts thereof are intended to be included within the scope of the present disclosure.
  • some of the compounds of the present disclosure may form solvates with water (i.e., a hydrate) or common organic solvents.
  • Such solvates and hydrates, particularly the pharmaceutically acceptable solvates and hydrates, of the compounds of this disclosure are likewise encompassed within the scope of the compounds defined by Formula I and the pharmaceutically acceptable salts thereof, along with un-solvated and anhydrous forms of such compounds.
  • the compounds of Formula I or salts thereof including pharmaceutically acceptable salts thereof, embodiments thereof and specific compounds described and claimed herein encompass all possible stereoisomers, tautomers, physical forms (e.g., amorphous and crystalline forms), co-crystal forms, solvate and hydrate forms, and any combination of the foregoing forms where such forms are possible.
  • Another embodiment of the present disclosure is a compound of Formula I wherein the compound or its salt is in a substantially pure form.
  • substantially pure means suitably at least about 60 wt.%, typically at least about 70 wt.%, preferably at least about 80 wt.%, more preferably at least about 90 wt.% (e.g., from about 90 wt.% to about 99 wt.%), even more preferably at least about 95 wt.% (e.g., from about 95 wt.% to about 99 wt.%, or from about 98 wt.% to 100 wt.%), and most preferably at least about 99 wt.% (e.g., 100 wt.%) of a product containing a compound of Formula I or its salt (e.g., the product isolated from a reaction mixture affording the compound or salt) consists of the compound or salt.
  • a product containing a compound of Formula I or its salt e.g., the product isolated from a
  • the level of purity of the compounds and salts can be determined using a standard method of analysis such as, high 25700 performance liquid chromatography, and/or mass spectrometry or NMR techniques. If more than one method of analysis is employed and the methods provide experimentally significant differences in the level of purity determined, then the method providing the highest purity level governs.
  • a compound or salt of 100% purity is one which is free of detectable impurities as determined by a standard method of analysis.
  • a substantially pure compound can be either a substantially pure mixture of the stereoisomers or a substantially pure individual stereoisomer.
  • the compounds of Formula I herein, and pharmaceutically acceptable salts thereof are useful for the inhibition of respiratory syncytial virus replication and metapneumovirus replication.
  • the compounds of Formula I and pharmaceutically acceptable salts thereof are useful for: (i) a method for the treatment of respiratory syncytial virus infection in a human subject in need thereof which comprises administering to the human subject an effective amount of the compound according to Formula I, or a pharmaceutically acceptable salt thereof; (ii) a method for the prophylaxis of respiratory syncytial virus infection in a human subject in need thereof which comprises administering to the human subject an effective amount of the compound according to Formula I, or a pharmaceutically acceptable salt thereof; (iii) a method for the treatment of metapneumovirus infection in a human subject in need thereof which comprises administering to the human subject an effective amount of the compound according to Formula I, or a pharmaceutically acceptable salt thereof; and/or (iv) a method for the prophylaxis of metapneumovirus infection in
  • any of the disclosed compounds in therapy are uses of any of the disclosed compounds as a medicament, or in the manufacture of a medicament, for the treatment of respiratory syncytial virus infection, prophylaxis of respiratory syncytial virus infection, treatment of metapneumovirus infection, prophylaxis of metapneumovirus infection, or inhibition of the replication of hRSV or hMPV.
  • Additional embodiments of the present disclosure include the following: (a) a pharmaceutical composition comprising an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, and 25700 (b) a pharmaceutical composition which comprises the product prepared by combining (e.g., mixing) an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. [0047] Additional embodiments of the present disclosure include each of the pharmaceutical compositions, methods and uses set forth in the preceding paragraphs, wherein the compound of Formula I or its salt employed therein in substantially pure.
  • composition comprising a compound of Formula I or its salt and a pharmaceutically acceptable carrier and optionally one or more excipients
  • administration and variants thereof (e.g., “administering” a compound) in reference to a compound of Formula I means providing the compound to the individual in need of treatment or prophylaxis and includes both self-administration and administration to the patient by another person or any other means.
  • composition is intended to encompass a product comprising the specified ingredients, as well as any product which results from combining the specified ingredients.
  • ingredients suitable for inclusion in a pharmaceutical composition are pharmaceutically acceptable ingredients, which means the ingredients must be compatible with each other and not rious to the recipient thereof.
  • a “subject” refers to an animal, such as a human, that is the object of treatment, observation or experiment.
  • a “subject” encompasses a mammalian animal.
  • the subject encompasses a domesticated or companion animal, or an experimental animal model.
  • the subject is a rodent, such as a mouse or rat.
  • the subject is a primate.
  • the subject is a non-human primate, such as a macaque.
  • the subject is a human.
  • human subject or “patient” as used herein refers to a human (or “person”) who has been the object of treatment, observation or experiment.
  • Patients to be treated with an RSV inhibitor (RSV-i) and/or an MPV inhibitor (MPV-i) agent include but are not limited to, patients who have been infected with RSV and/or MPV.
  • Patients to be treated with an RSV-i and/or an MPV-i agent also include, but are not limited to, those using an RSV-i and/or an MPV- i agent for prophylaxis of RSV and/or MPV infection or for post-exposure prophylaxis after being potentially exposed to RSV and/or MPV to prevent or reduce the severity of symptoms of virus-associated disease or condition.
  • “Prophylaxis” includes each of (i) pre-exposure prophylaxis (PrEP), i.e., using a compound of Formula I, or a pharmaceutically acceptable salt thereof, to prevent hRSV and/or hMPV infection in a person who is not infected with hRSV and/or hMPV, and (ii) post-exposure prophylaxis (PEP), i.e., using a compound of Formula I, or a pharmaceutically acceptable salt thereof, after being exposed or potentially exposed to hRSV and/or hMPV to prevent or reduce the severity of symptoms of virus-associated disease or condition.
  • PrEP pre-exposure prophylaxis
  • PEP post-exposure prophylaxis
  • a compound of Formula I, or a pharmaceutically acceptable salt thereof is administered for PEP in patients exposed to RSV to minimize the risk of severe RSV disease or symptoms thereof.
  • the term “effective amount” as used herein means the amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician.
  • the effective amount is a “therapeutically effective amount” for the alleviation of the symptoms of the disease or condition being treated.
  • the effective amount is a “prophylactically effective amount” for prophylaxis of the symptoms of the disease or condition being prevented.
  • references to the amount of active ingredient are to the free form (i.e., the non-salt form) of the compound.
  • the disclosed compounds, and salts thereof can be administered by means that produce contact of the active agent with the agent’s site of action. They can be administered by conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or with other therapeutic agents the patient may be in need of.
  • the compound can be administered itself, but typically is administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.
  • the compounds of the invention can, for example, be administered orally (e.g., via tablet or capsule), parenterally (including subcutaneous injection; intravenous, intramuscular or intrasternal injection; or infusion techniques), by inhalation spray, or rectally, in the form of a unit dosage of a pharmaceutical composition containing an effective amount of the compound and conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles.
  • the compound could also be administered via an implantable drug delivery device adapted to provide an effective amount of the compound or a pharmaceutical composition of the compound over an extended period of time.
  • Solid preparations suitable for oral administration can be prepared according to techniques known in the art and can employ such solid excipients as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like.
  • Liquid preparations suitable for oral administration e.g., suspensions, syrups, elixirs and the like
  • Parenteral compositions can be prepared according to techniques known in the art and typically employ sterile water as a carrier and optionally other ingredients, such as a solubility aid.
  • Injectable solutions can be prepared according to methods known in the art wherein the carrier comprises a saline solution, a glucose solution or a solution containing a mixture of saline and glucose.
  • Implantable compositions can be prepared according to methods known in the art wherein the carrier comprises the active chemical ingredient with polymers and suitable excipients, or utilizing an implantable device for drug delivery. Further description of methods suitable for use in preparing pharmaceutical compositions for use in the present disclosure and of ingredients suitable for use in said compositions is provided in Remington - The Science and Practice of Pharmacy, 22nd Edition, published by Pharmaceutical Press and Philadelphia College of Pharmacy at University of the Sciences, 2012, ISBN 978085711-062-6 and prior editions.
  • Formulations of compounds of Formula I that result in drug supersaturation and/or rapid dissolution may be utilized to facilitate oral drug absorption.
  • Formulation approaches to cause drug supersaturation and/or rapid dissolution include, but are not limited to, nanoparticulate systems, amorphous systems, solid solutions, solid dispersions, and lipid systems.
  • Such formulation approaches and techniques for preparing them are known in the art.
  • solid dispersions can be prepared using excipients and processes as described in reviews (e.g., A.T.M. Serajuddin, J Pharm Sci, 88:10, pp.1058-1066 (1999)).
  • Nanoparticulate systems based on both attrition and direct synthesis have also been described in reviews such as Wu et al. (F.
  • the compounds of Formula I may be administered in a dosage range of, e.g., 1 to 20 mg/kg, or 1 to 10 mg/kg, or about 5 mg/kg of mammal (e.g., human) body weight per day, or at other time intervals as appropriate, in a single dose or in divided doses.
  • the compounds of Formula I may be administered in a dosage range of 0.001 to 2000 mg per day in a single dose or in divided doses.
  • dosage ranges are 0.01 to 1500 mg per day, or 0.1 to 1000 mg per day, administered orally or via other routes of administration in a single dose or in divided doses. 25700 [0059]
  • the dosage units may contain 100 mg to 1500 mg of the active ingredient, for example but not limited to 0.1 mg to about 1500 mg of the active ingredient, for example but not limited to 0.1, 0.25, 0.5, 1, 2, 2.5, 5, 10, 15, 20, 25, 40, 50, 75, 100, 125, 150, 175, 200, 225, 250, 500, 1000, 1250, or 1500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated.
  • the compound may be formulated in oral formulations for immediate or modified release such as extended or controlled release.
  • immediate or modified release such as extended or controlled release.
  • reference to an amount of the compound in milligrams or grams is based on the free form (i.e., the non-salt form) of the compound.
  • Daily administration can be via any suitable route of administration but is preferably via oral administration and can be a single dose or more than one dose at staggered times (divided daily doses) within each 24-hour period. Each dose may be administered using one or multiple dosage units as appropriate.
  • the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, the effect of other drugs the patient is taking while using and RSP-i or an MPV-i compound described herein, the severity of the particular condition, and the host undergoing therapy. In some cases, depending on the potency of the compound or the individual response, it may be necessary to deviate upwards or downwards from the given dose. The amount and frequency of administration will be regulated according to the judgment of the attending clinician considering such factors.
  • the compounds of this invention are also useful in the preparation and execution of screening assays for antiviral compounds.
  • the compounds of this invention are useful for isolating enzyme mutants, which are excellent screening tools for more powerful antiviral compounds.
  • the compounds of this invention are useful in establishing or determining the binding site of other antivirals.
  • a chiral center in a compound may exist in the "S" or "R" stereo- configuration, or as a mixture of both. Within a molecule, each bond drawn as a straight line from a chiral center encompasses each of the (R) and (S) stereoisomers as well as mixtures thereof unless otherwise noted.
  • isomer mixtures may have been separated, providing one or both of an isomer 1 (the faster eluting isomer) and an isomer 2 (the slower eluting isomer), based on their observed elution order resulting from the separation as performed in the Example.
  • Elution time and/or order of separated 25700 isomers may differ if performed under conditions different than those employed herein.
  • Absolute stereochemistry (R or S) of the chiral center in each of isomer “1” and/or isomer “2” separated stereoisomers in the intermediates and Examples was not determined, and “1” and “2” only refer to elution order resulting from the purification conditions as performed.
  • compounds of Formula (I) can be prepared by amide coupling of appropriately functionalized carboxylic acids of Formula (II) and amines of Formula (III). Acids of Formula (II) and amines of Formula (III) are commercially available or may be synthesized from appropriate intermediates. Other synthetic methods to prepare compounds of Formula (I) 25700 are presented for the exemplification of compounds. Further, compounds of Formula (I) can be prepared by reaction procedures generally known in the art.
  • the column used was a Waters Acquity BEH C181 ⁇ 50 mm, 1.7 ⁇ m, heated to 50 oC.
  • the mobile phases used were modified with either acidic or basic additives.
  • the acidic mobile phase consisted of water (with 0.1% TFA modifier) for Solvent A and 100% MeCN for Solvent B.
  • a two-minute run was established at a flow rate of 0.3 mL/min with initial conditions of 95% Solvent A and ramping up to 99% Solvent B at 1.60 minutes and holding at 99% Solvent B for 0.40 minutes.
  • the injection volume was 0.5 ⁇ L using partial loop needle overfill injection mode.
  • the TUV monitored wavelength 215 or 254 nm with a sampling rate of 20 points/second, normal filter constant and absorbance data mode.
  • the basic mobile phase consisted of water (with 0.05% NH 4 OH modifier) for solvent A and 100% MeCN for solvent B.
  • a two-minute run was established at a flow rate of 0.3 mL/min with Initial conditions of 99% Solvent A and ramping up to 99% Solvent B at 1.90 minutes and holding at 99% Solvent B for 0.10 minutes.
  • a five-minute run was established at a flow rate of 0.3 mL/min with initial conditions of 95% Solvent A and ramping up to 99% Solvent B at 4.90 minutes and holding at 99% Solvent B for 0.10 minutes.
  • the injection volume was 5.0 ⁇ L using Partial Loop Needle Overfill Injection mode.
  • the TUV monitored wavelength 215 nm with a sampling rate of 20 points/second, normal filter constant and absorbance data mode.
  • a commonly used system consisted of a Waters ZQTM platform with electrospray ionization in positive ion detection mode with an Agilent 1100 series HPLC with autosampler.
  • the column was commonly a Waters Xterra MS C18, 3.0 ⁇ 50 mm, 5 ⁇ m or a Waters Acquity UPLC® BEH C181.0 x 50 mm, 1.7 ⁇ m.
  • the flow rate was 1 mL/min, and the injection volume was 10 ⁇ L.
  • UV detection was in the range 210–400 nm.
  • the mobile phase consisted of solvent A (water with 0.05% TFA modifier) and solvent B (MeCN with 0.05% TFA modifier) with a gradient of 100% solvent A for 0.7 min changing to 100% solvent B over 3.75 min, maintained for 1.1 min, then reverting to 100% solvent A over 0.2 min.
  • Preparative reverse-phase chromatography was generally carried out on a Teledyne ISCO ACCQPrep HP125 or HP150 apparatus equipped with UV and ELSD detectors.
  • the UV detector typically monitored wavelengths of 215 and 254 nm.
  • the column was commonly one of the following: Waters XBridge Prep C18 OBD 5 ⁇ m 30 ⁇ 150 mm, Waters XBridge Prep C18 OBD 5 ⁇ m 30 ⁇ 250 mm, Waters XBridge Prep C18 OBD 5 ⁇ m 50 ⁇ 250 mm, Waters SunFire Prep C18 OBD 5 ⁇ m 30 ⁇ 150 mm, Waters SunFire Prep C18 OBD 10 ⁇ m 30 ⁇ 150 mm, Waters SunFire Prep C18 OBD 5 ⁇ m 50 ⁇ 250 mm, Waters SunFire Prep C18 OBD 10 ⁇ m 50 ⁇ 250 mm, or Phenomenex Luna Prep C185 ⁇ m 50 ⁇ 250 mm.
  • the mobile phases consisted of mixtures of MeCN (with 0.1% TFA modifier) and water (with 0.1% TFA modifier).
  • a commonly used system was a Waters Chromatography Workstation configured with an LCMS system consisting of: Waters ZQTM single quad MS system with Electrospray Ionization, Waters 2525 Gradient Pump, Waters 2767 Injector/Collector, Waters 996 PDA Detector.
  • MS conditions were: 150-750 amu, positive electrospray, collection triggered by MS.
  • Flow rates were maintained at 50 mL/min for the Waters XBridge column, 90 mL/min for the Phenomenex Gemini column, and 25 mL/min for the Welch Xtimate column.
  • the UV detection range was 210–400 nm.
  • Mobile phase gradients were optimized for the individual compounds.
  • Silica gel chromatography was usually performed using an ISCO CombiFlash Rf apparatus, a Biotage® Flash Chromatography apparatus (Dyax Corp.), or an ISCO CombiFlash® Companion XL apparatus on silica gel (60 ⁇ pore size) in pre-packed RediSep Rf, RediSep Rf Gold, or SepaFlash columns.
  • Chiral chromatography was commonly performed by supercritical fluid chromatography with a column chosen from one of the following: ChiralPak AD, ChiralPak AD-3, ChiralPak AD- H, ChiralPak AS, ChiralPak AS-3, ChiralPak AS-H, ChiralPak IB-N, ChiralPak OD-H, 25700 ChiralPak OJ-3, ChiralPak OJ-H, Phenomenex-Cellulose-2, or (S,S)Whelk-O1.
  • Mobile phases consisted of mixtures of CO2 or hexane with MeOH, EtOH, or IPA using 0.05-0.1% DEA or NH 4 OH modifier. Mobile phase gradients were optimized for the individual compounds. Pressure was typically maintained at 100 bar, and flow rates ranged from 50-200 mL/min. UV monitoring was generally carried out at 220 or 205 nM.
  • 1 H NMR data were typically acquired using a Bruker NEO 500 MHz NMR spectrometer equipped with a room temperature 5 mm BBF iProbe, a Bruker Avance NEO 400 MHz NMR spectrometer equipped with a Bruker PI HR-BBO400S1-BBF/H/D-5.0-Z SP probe, or a Bruker Avance III 500 MHz NMR spectrometer equipped with a Bruker 5mm PABBO probe. Chemical shift values are reported in delta ( ⁇ ) units, parts per million (ppm).
  • Lithium bis(trimethylsilyl)amide solution (1.0 M, 1.5 L, 1.5 mol) in isopropyl ether (3.0 L) was charged into a 10 L vessel.
  • Diiodomethane 130 mL, 1.6 mol was added to the mixture at –65 °C.
  • benzylchlorodimethylsilane 200 g, 1.1 mol, 200 mL was added.
  • 3 h the mixture was warmed to rt.
  • After 12 h the reactions were combined and treated with water (10 L).
  • the resulting mixture was extracted with MTBE (5 L x 2).
  • the combined organic extracts were washed with brine then dried (Na 2 SO 4 ) then filtered then the filtrate was concentrated.
  • Step 2 tert-butyl (1R,5S,6s)-6-(benzyldimethylsilyl)-3-azabicyclo[3.1.0]hexane-3- carboxylate [0074] Fourteen reactions were carried out in parallel. Chromium (II) chloride (150 g, 1.2 mol) was charged to a 5 L vessel. THF (2.5 L) and N,N,N,N-tetramethylethylenediamine (360 mL, 2.4 mol) were added to the mixture at rt over 30 minutes.
  • Chromium (II) chloride 150 g, 1.2 mol
  • THF 2.5 L
  • N,N,N,N-tetramethylethylenediamine 360 mL, 2.4 mol
  • Benzyl(diiodomethyl)dimethylsilane (240 g, 560 mmol) was added and the mixture was stirred at rt for 30 minutes.
  • tert-Butyl 2,5-dihydro- 1H-pyrrole-1-carboxylate (50 g, 0.30 mol) was added then the mixture was heated to 50 °C. After 18 h the mixture was cooled to rt then the reactions were combined and treated with water (15 L). The resulting mixture was extracted with MTBE (7 L x 3). The combined organic extracts were washed with brine then dried (Na2SO4) then filtered then the filtrate was concentrated.
  • Tetrabutylammonium fluoride solution (1.0M in THF, 630 mL) was added at rt. After 30 minutes the mixture was cooled to 0 °C then MeOH (2.1 L) and K2CO3 (200 g, 2.1 mol) were added followed by hydrogen peroxide (30 wt % in water, 100 mL, 1.1 mol). After 30 minutes the mixture was warmed to rt. After 16 h the reactions were combined then treated with Na 2 SO 3 (saturated aq., 25 L). The resulting mixture was extracted with EtOAc (7 L x 2). The combined organic extracts were washed with brine then dried (Na2SO4) then filtered then the filtrate was concentrated.
  • Step 2 benzyl (1R,5S,6s)-6-hydroxy-3-azabicyclo[3.1.0]hexane-3-carboxylate [0077] To a solution of (1R,5S,6s)-3-azabicyclo[3.1.0]hexan-6-ol TFA salt (245 mg, 2.47 mmol) and K 2 CO 3 (520 mg, 3.71 mmol) in THF (10 mL) was added benzyl chloroformate (0.52 mL, 3.71 mmol) at 0 oC.
  • Step 2 tert-butyl 6-hydroxy-1-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylate
  • a solution of tert-butyl 6,6-dibromo-1-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylate (2.0 g, 5.6 mmol) in THF (40 mL) was cooled to –78 °C.
  • Step 2 benzyl (2-(2-chloropyridin-4-yl)propan-2-yl)carbamate
  • 2-(2-chloropyridin-4-yl)propan-2-amine 64.1 g, 376 mmol
  • DIEA 79 mL, 451 mmol
  • benzyl chloroformate 80 mL, 563 mmol
  • the resulting mixture was diluted with water (150 mL) then extracted with DCM (150 mL x 3).
  • Step 3 benzyl (2-(2-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate
  • Step 4 4-(2-(((benzyloxy)carbonyl)amino)propan-2-yl)-2-(4-fluorophenyl)pyridine 1-oxide
  • 3-Chloroperoxybenzoic acid (18.83 g, 93 mmol) was added to a solution of benzyl (2- (2-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate (16.9 g, 46.4 mmol) in DCM (300 mL) at rt.
  • Step 2 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)propan-2-amine
  • Step 3 benzyl (2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate
  • 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)propan-2-amine 500 g, 1.89 mol
  • DCM 3.0 L
  • DIPEA 269 g, 2.08 mol
  • benzyl chloroformate 354 g, 2.08 mol
  • Step 2 tert-butyl rac-2-(2,6-dichloropyridin-4-yl)-2-methylazetidine-1-carboxylate
  • tert-butyl rac-2-(2,6-dichloropyridin-4-yl)azetidine-1-carboxylate 240 mg, 0.79 mmol
  • THF 4.0 mL
  • Step 3 tert-butyl 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-2-methylazetidine-1- carboxylate
  • tert-butyl 2-(2,6-dichloropyridin-4-yl)-2-methylazetidine-1-carboxylate 110 mg, 0.35 mmol
  • (4-fluorophenyl)boronic acid 54 mg, 0.39 mmol
  • Pd(dppf)Cl2-DCM adduct 15 mg, 0.018 mmol
  • Step 2 rac-4-(1-((benzyloxy)carbonyl)-2-methylpyrrolidin-2-yl)pyridine 1-oxide
  • benzyl rac-2-methyl-2-(pyridin-4-yl)pyrrolidine-1-carboxylate 510 mg, 1.7 mmol
  • 3-chloroperoxybenzoic acid 640 mg, 2.6 mmol
  • 25700 After 3 h the mixture was concentrated.
  • the crude product was subjected to silica gel chromatography (0-100% (3:1 EtOAc:EtOH)/heptane) to give the title compound.
  • Step 3 rac-4-(1-((benzyloxy)carbonyl)-2-methylpyrrolidin-2-yl)-2-(4-fluorophenyl)pyridine 1-oxide
  • 1-Bromo-4-fluorobenzene (0.13 mL, 1.2 mmol)
  • rac-4-(1-((benzyloxy)carbonyl)-2- methylpyrrolidin-2-yl)pyridine 1-oxide 440 mg, 1.4 mmol
  • K 2 CO 3 330 mg, 2.4 mmol
  • Step 4 benzyl rac-2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-2-methylpyrrolidine-1- carboxylate
  • a solution of rac-4-(1-((benzyloxy)carbonyl)-2-methylpyrrolidin-2-yl)-2-(4- fluorophenyl)pyridine 1-oxide 190 mg, 0.47 mmol
  • DCM DCM
  • triethylamine 0.27 mL, 1.9 mmol
  • oxalyl chloride 0.085 mL, 0.97 mmol
  • Step 2 6-chloro-4-(4-fluorophenyl)picolinonitrile
  • a solution of 2-chloro-4-(4-fluorophenyl)pyridine 1-oxide (4.3 g, 19 mmol), trimethylsilyl cyanide (11 g, 0.12 mol) and triethylamine (5.4 mL, 39 mmol) was heated under an atmosphere of nitrogen at 100 °C for 12 h. The mixture was cooled to rt then concentrated. The crude product was subjected to silica gel chromatography (0-40% EtOAc/petroleum ether) to give the title compound.
  • Step 3 2-(6-chloro-4-(4-fluorophenyl)pyridin-2-yl)propan-2-amine [0097] To a solution of 6-chloro-4-(4-fluorophenyl)picolinonitrile (2.7 g, 12 mmol) in THF (80 mL) was slowly added methylmagnesium bromide (3.0 M in Et 2 O, 19 mL, 58 mmol) at 0 °C. After the addition was complete the mixture was warmed to rt. After 30 minutes titanium isopropoxide (3.4 mL, 12 mmol) was added then the mixture was heated to 80 °C.
  • methylmagnesium bromide 3.0 M in Et 2 O, 19 mL, 58 mmol
  • Step 4 benzyl (2-(6-chloro-4-(4-fluorophenyl)pyridin-2-yl)propan-2-yl)carbamate
  • 2-(6-chloro-4-(4-fluorophenyl)pyridin-2-yl)propan-2-amine 2.0 g, 7.6 mmol
  • DIPEA 4.0 mL, 23 mmol
  • benzyl chloroformate 2.2 mL, 15 mmol
  • Step 4 2-(2,6-dichloropyridin-4-yl)propan-2-amine [0102] A solution of 2,6-dichloroisonicotinonitrile (300 g, 1.73 mol) in toluene (1.5 L) was cooled to 0 °C. To this was added methylmagnesium bromide solution (3M in diethyl ether, 1.73 L, 5.20 mol) dropwise over 1 h.
  • Step 5 benzyl (2-(2,6-dichloropyridin-4-yl)propan-2-yl)carbamate
  • 2-(2,6-dichloropyridin-4-yl)propan-2-amine 270 g, 1.32 mol
  • DIPEA 187 g, 1.45 mol
  • benzyl chloroformate 247 g, 1.45 mol
  • the mixture was warmed to rt. After 12 h the mixture was washed with 1N HCl (800 mL x 3) then dried (Na 2 SO 4 ) then filtered then the filtrate was concentrated.
  • Step 2 3-fluoro-2-(4-fluorophenyl)-4-(methoxycarbonyl)pyridine 1-oxide
  • 3-Chloroperoxybenzoic acid (2.44 g, 12.04 mmol) was added to a solution of methyl 3- fluoro-2-(4-fluorophenyl)isonicotinate (1.5 g, 6.02 mmol) in DCM (40 mL) at rt. After 12 h the mixture was diluted with saturated Na 2 SO 3 (15 mL) and saturated aqueous NaHCO 3 (15 mL) then extracted with DCM (3 x 20 mL).
  • Step 5 2-chloro-N-(2-(6-chloro-3-fluoro-2-(4-fluorophenyl)pyridin-4-yl)propan-2- yl)acetamide
  • 2-(6-chloro-3-fluoro-2-(4-fluorophenyl)pyridin-4-yl)propan-2-ol 2.6 g, 9.16 mmol
  • 2-chloroacetonitrile 20 mL, 316 mmol
  • Step 6 2-(6-chloro-3-fluoro-2-(4-fluorophenyl)pyridin-4-yl)propan-2-amine [0110] To a solution of 2-chloro-N-(2-(6-chloro-3-fluoro-2-(4-fluorophenyl)pyridin-4- yl)propan-2-yl)acetamide (1.65 g, 4.59 mmol) in MeOH (30 mL) and AcOH (5 mL) was added thiourea (0.699 g, 9.19 mmol) then the mixture was heated to 80 °C for 12 h. The mixture was cooled to rt then concentrated.
  • Step 7 benzyl (2-(6-chloro-3-fluoro-2-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate [0111] To a solution of 2-(6-chloro-3-fluoro-2-(4-fluorophenyl)pyridin-4-yl)propan-2-amine (50 mg, 0.18 mmol) in DCM (2 mL) was added DIPEA (0.154 mL, 0.88 mmol) then benzyl chloroformate (0.037 mL, 0.265 mmol) at rt.
  • Step 2 methyl 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)propanoate [0113] Under an atmosphere of nitrogen, to a –78 °C solution of methyl 2-(2-chloro-6-(4- fluorophenyl)pyridin-4-yl)acetate (14 g, 0.050 mol) in THF (150 mL) was added a solution of lithium bis(trimethylsilyl)amide (1.0M in THF, 0.060 L, 0.060 mol).
  • Step 3 methyl 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-3-hydroxy-2-methylpropanoate
  • aqueous formaldehyde (2.76 g, 34.0 mmol, 37% in water) at rt under N2.
  • Step 4 methyl 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-3-fluoro-2-methylpropanoate [0115] To a solution of methyl 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-3-hydroxy-2- methylpropanoate (1 g, 4.35 mmol) in DCM (20 mL) was added DAST (5.8 mL, 43.5 mmol) dropwise at -78 °C under N2. The mixture was slowly warmed to rt over 16 h.
  • Step 5 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-3-fluoro-2-methylpropanoic acid [0116] To a mixture of methyl 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-3-fluoro-2- methylpropanoate (300 mg, 0.92 mmol) in THF (5 mL) and H2O (5 mL) was added lithium hydroxide (110 mg, 4.60 mmol) at rt. After 2 h the mixture was concentrated to remove volatile organic solvents. The resulting mixture was washed with DCM (10 mL x 2).
  • the aqueous phase was adjusted to pH 5 using 1M HCl then concentrated.
  • the crude product was taken up in 20 mL 1:1 MeOH:DCM and stirred at rt for 1 h. The resulting mixture was filtered then the filtrate was concentrated in vacuo to give the title compound.
  • Step 6 benzyl (2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-1-fluoropropan-2-yl)carbamate
  • 2-(2-chloro-6-(4-fluorophenyl)pyridin-4-yl)-3-fluoro-2- methylpropanoic acid 270 mg, 0.87 mmol
  • DIPEA 0.6 mL, 3.46 mmol
  • DPPA 0.3 mL, 1.732 mmol
  • benzyl alcohol 187 mg, 1.73 mmol
  • Step 2 benzyl (2-(2-chloro-6-(((1R,5S,6s)-3-(2-methyl-8-(trifluoromethyl)imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2- yl)carbamate
  • 2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carboxylic acid 0.478 g, 1.96 mmol
  • HATU (1.12 g, 2.94 mmol
  • DIPEA 1, 7.71 mL, 9.79 mmol
  • Step 2 benzyl (2-(2-(((1R,5S,6s)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4- fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate
  • Step 2 tert-butyl (2-(2-(((1R,5S,6s)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4- fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate [0127] To a solution of benzyl (1R,5S,6s)-6-((4-(2-((tert-butoxycarbonyl)amino)propan-2-yl)- 6-(4-fluorophenyl)pyridin-2-yl)oxy)-3-azabicyclo[3.1.0]hexane-3-carboxylate (500 mg, 0.890 mmol) in THF (20 mL) was added TEA (450 mg, 4.45 mmol), triethylsilane (414 mg, 3.56 m
  • Step 2 benzyl rac-(2-(2-(4-fluorophenyl)-6-(((1R,5S,6S)-1-methyl-3-azabicyclo[3.1.0]hexan- 6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate [0129] To a mixture of tert-butyl rac-(1R,5S,6S)-6-((4-(2- (((benzyloxy)carbonyl)amino)propan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-1-methyl-3- azabicyclo[3.1.0]hexane-3-carboxylate (150 mg, 0.26 mmol) in DCM (4.0 mL) was added TFA (1.0 mL) at rt.
  • Step 2 benzyl (2-(2-(((1R,5S,6s)-3-azabicyclo[3.1.0]hexan-6-yl)amino)-6-(4- fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate [0131] To a solution of tert-butyl (1R,5S,6s)-6-((4-(2-(((benzyloxy)carbonyl)amino)propan-2- yl)-6-(4-fluorophenyl)pyridin-2-yl)amino)-3-azabicyclo[3.1.0]hexane-3-carboxylate (536 mg, 0.96 mmol) in DCM (5 mL) was added TFA (2 mL) at rt.
  • the reaction was quenched by addition of 2M HCl (2 mL) maintaining temperature below 25° C.
  • the 25700 resulting mixture was diluted with water (10 mL) then extracted with EtOAc (3 x 10 mL).
  • the combined organic extracts were washed with water (3 x 10 mL) then dried (MgSO4) then filtered then the filtrate was concentrated.
  • the crude product was subjected to silica gel chromatography (0-10% EtOAc/petroleum ether) to give the title compound.
  • Step 2 tert-butyl (1R,5S,6s)-6-((5-cyano-4'-fluoro-[1,1'-biphenyl]-3-yl)oxy)-3- azabicyclo[3.1.0]hexane-3-carboxylate
  • Step 3 tert-butyl (1R,5S,6s)-6-((5-(2-aminopropan-2-yl)-4'-fluoro-[1,1'-biphenyl]-3-yl)oxy)- 3-azabicyclo[3.1.0]hexane-3-carboxylate
  • a 20 mL Biotage microwave process tube with stir bar was charged with tert-butyl (1R,5S,6s)-6-((5-cyano-4'-fluoro-[1,1'-biphenyl]-3-yl)oxy)-3-azabicyclo[3.1.0]hexane-3- carboxylate (690 mg, 1.75 mmol) and THF (8 mL).
  • methylmagnesium bromide 3M in diethyl ether, 2.33 mL, 7.00 mmol
  • Ti(OiPr)4 (0.53 mL, 1.75 mmol) was added then the mixture was heated by microwave irradiation at 50 °C for 1 h.
  • the resulting mixture was quenched with a saturated Na2CO3 (25 mL) then filtered through a pad of Celite ® washing with DCM (3 x 50 mL).
  • Step 4 tert-butyl (1R,5S,6s)-6-((5-(2-(((benzyloxy)carbonyl)amino)propan-2-yl)-4'-fluoro- [1,1'-biphenyl]-3-yl)oxy)-3-azabicyclo[3.1.0]hexane-3-carboxylate [0136] To a solution of tert-butyl (1R,5S,6s)-6-((5-(2-aminopropan-2-yl)-4'-fluoro-[1,1'- biphenyl]-3-yl)oxy)-3-azabicyclo[3.1.0]hexane-3-carboxylate (680 mg, 1.59 mmol) and DIPEA (0.84 mL, 4.78
  • Step 2 tert-butyl rac-(1R,5S,6s)-6-((4-(1-(((benzyloxy)carbonyl)amino)ethyl)-6-(4- fluorophenyl)pyridin-2-yl)oxy)-3-azabicyclo[3.1.0]hexane-3-carboxylate
  • a 5 mL Biotage microwave process tube with a magnetic stir bar was charged with tert- butyl (1R,5S,6s)-6-((4-cyano-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3-azabicyclo[3.1.0]hexane-3- carboxylate (50 mg, 0.13 mmol) and THF (2 mL).
  • Step 3 tert-butyl ent-(1R,5S,6s)-6-((4-(1-(((benzyloxy)carbonyl)amino)ethyl)-6-(4- fluorophenyl)pyridin-2-yl)oxy)-3-azabicyclo[3.1.0]hexane-3-carboxylate (enantiomers 1 and 2) [0141]
  • the racemic mixture from step 2 was subjected to chiral SFC (Chiralpak IC-3, 40% IPA/CO2) to give the title compounds, Int.
  • Step 2 methyl 2-cyclopropyl-8-methoxyimidazo[1,2-a]pyridine-6-carboxylate
  • Methyl 8-bromo-2-cyclopropylimidazo[1,2-a]pyridine-6-carboxylate 200 mg, 0.68 mmol
  • Cs2CO3 442 mg, 1.36 mmol
  • methanol 65.1 mg, 2.03 mmol
  • Step 2 methyl 8-(2,4-dioxoimidazolidin-1-yl)-2-methylimidazo[1,2-a]pyridine-6- carboxylate
  • 2-chloroacetyl isocyanate 29.1 mg, 0.244 mmol
  • DBU 0.257 ml, 1.71 mmol
  • Step 3 8-(2,4-dioxoimidazolidin-1-yl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • THF 2 mL
  • potassium trimethylsilanolate 56 mg, 0.434 mmol
  • Step 2 2-methyl-8-(1H-tetrazol-1-yl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • methyl 2-methyl-8-(1H-tetrazol-1-yl)imidazo[1,2-a]pyridine-6- carboxylate 30 mg, 0.12 mmol
  • water 0.5 mL
  • potassium trimethylsilanolate 75 mg, 0.58 mmol
  • 3 h H2O 3 mL
  • 1M HCl was used to adjust pH of the mixture to 3.
  • the resulting mixture was washed with EtOAc then the water layer was lyophilized to give the title compound.
  • Step 2 methyl 8-((2-aminoethyl)amino)-2-methylimidazo[1,2-a]pyridine-6-carboxylate 25700 [0157] To a solution of methyl 8-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-2- methylimidazo[1,2-a]pyridine-6-carboxylate (180 mg, 0.52 mmol) in DCM (6 mL) was added TFA (3 mL) at rt.
  • Step 3 methyl 2-methyl-8-(2-oxoimidazolidin-1-yl)imidazo[1,2-a]pyridine-6-carboxylate
  • DIPEA 1.2 mL, 6.85 mmol
  • CDI 167 mg, 1.03 mmol
  • Step 4 2-methyl-8-(2-oxoimidazolidin-1-yl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • a mixture of methyl 2-methyl-8-(2-oxoimidazolidin-1-yl)imidazo[1,2-a]pyridine-6- carboxylate (160 mg, 0.58 mmol) in THF (5 mL) and water (5 mL) was added LiOH (41.9 mg, 1.75 mmol) at rt. After 2 h the mixture was concentrated to remove volatile organic solvents.
  • Step 2 8-cyclopropyl-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid [0161] To a solution of ethyl 8-cyclopropyl-2-methylimidazo[1,2-a]pyridine-6-carboxylate (1.01 g, 4.13 mmol) in MeOH (21 mL) was added 1M NaOH (4.13 mL, 4.13 mmol) at rt. After 16 h the mixture was concentrated to remove volatile organic solvents. MeCN (10 mL) was added then the mixture was lyophilized to give the sodium salt of the title compound.
  • Step 2 ethyl 2-chloro-8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carboxylate
  • a mixture of ethyl 2-hydroxy-8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carboxylate (300 mg, 0.66 mmol) and POCl3 (0.306 mL, 3.28 mmol) in DCE (10 mL) was heated to 80 °C. After 12 h the mixture was cooled to rt then concentrated. The residue was taken up in saturated NaHCO3then the mixture was extracted with DCM (30 mL x 2).
  • Step 3 2-chloro-8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carboxylic acid [0165] To a mixture of ethyl 2-chloro-8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carboxylate (51 mg, 0.174 mmol) in THF (5 mL) and water (3 mL) was added NaOH (34.9 mg, 0.871 mmol) at rt. After 1 h the mixture was concentrated to remove volatile organic solvents. The resulting aqueous mixture was washed with DCM (20 mL x 2) then the pH was adjusted to 6 using 1M HCl.
  • Step 2 2-cyclopropyl-8-(difluoromethoxy)imidazo[1,2-a]pyridine-6-carboxylic acid
  • 6-bromo-2-cyclopropyl-8-(difluoromethoxy)imidazo[1,2-a]pyridine 7 g, 23.09 mmol
  • 1,3-bis(dicyclohexylphosphino)propane bis(tetrafluoroborate) 1.14 g, 2.31 mmol
  • K2CO3 5.75 g, 41.6 mmol
  • Step 2 2-(1-fluoroethyl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • 6-bromo-2-(1-fluoroethyl)imidazo[1,2-a]pyridine 50 mg, 0.21 mmol
  • 1,3-bis(dicyclohexylphosphino)propane bis(tetrafluoroborate) (12.59 mg, 0.021 mmol)
  • K 2 CO 3 51.2 mg, 0.37 mmol
  • DMSO 5 mL
  • water 0.5 mL
  • Pd(OAc) 2 2.3 mg, 10.28 ⁇ mol
  • Step 2 methyl 2-cyclopropyl-8-formylimidazo[1,2-a]pyridine-6-carboxylate
  • a solution of methyl 2-cyclopropyl-8-vinylimidazo[1,2-a]pyridine-6-carboxylate (350 mg, 1.45 mmol) and NMO (338 mg, 2.89 mmol) in THF (10 mL) was added a solution of osmium tetroxide (7.35 mg, 0.029 mmol) in water (0.5 mL) at rt. After 1 h sodium periodate (927 mg, 4.33 mmol) was added.
  • Step 3 methyl 2-cyclopropyl-8-(difluoromethyl)imidazo[1,2-a]pyridine-6-carboxylate [0173] To a solution of methyl 2-cyclopropyl-8-formylimidazo[1,2-a]pyridine-6-carboxylate (210 mg, 0.860 mmol) in DCM (6 mL) was added DAST (0.568 mL, 4.30 mmol) at -78 °C. The mixture as allowed to warm to rt over 12 h. The resulting mixture was diluted with saturated 25700 NaHCO 3 (30 mL) then extracted with DCM (20 mL x 2).
  • Step 4 2-cyclopropyl-8-(difluoromethyl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • THF methyl 2-cyclopropyl-8-(difluoromethyl)imidazo[1,2-a]pyridine-6- carboxylate
  • NaOH 128 mg, 3.19 mmol
  • the resulting aqueous mixture was washed with DCM (20 mL x 2) then the pH was adjusted to 5 using 1M HCl.
  • Step 1 ethyl 8-(2-hydroxyethyl)- pyridine-6-carboxylate
  • 9-BBN 0.5 M in THF, 1.72 mL, 0.86 mmol
  • ethyl 2-methyl- 8-vinylimidazo[1,2-a]pyridine-6-carboxylate 66 mg, 0.29 mmol
  • 16 h H2O2 30 wt% in water, 0.15 mL, 1.43 mmol
  • 1N NaOH (1.43 mL, 1.43 mmol
  • Step 2 8-(2-hydroxyethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • ethyl 8-(2-hydroxyethyl)-2-methylimidazo[1,2-a]pyridine-6- carboxylate 51.8 mg, 0.21 mmol
  • MeOH 2.1 mL
  • 1M NaOH 0.21 mL, 0.21 mmol
  • Step 2 2-methyl-8-(2,2,2-trifluoro-1-hydroxyethyl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • methyl 2-methyl-8-(2,2,2-trifluoro-1-hydroxyethyl)imidazo[1,2- a]pyridine-6-carboxylate 70 mg, 0.243 mmol
  • MeOH MeOH
  • water 3 mL
  • sodium hydroxide 29.1 mg, 0.729 mmol
  • Step 2 methyl 8-(2-amino-1-hydroxyethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate [0181] To a solution of methyl 8-(1-hydroxy-2-nitroethyl)-2-methylimidazo[1,2-a]pyridine-6- carboxylate (75 mg, 0.27 mmol) in MeOH (4 mL) was added Raney-Ni (16 mg, 0.27 mmol) under N 2 atmosphere.
  • Step 4 2-methyl-8-(2-oxooxazolidin-5-yl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • methyl 2-methyl-8-(2-oxooxazolidin-5-yl)imidazo[1,2-a]pyridine-6- carboxylate 35 mg, 0.13 mmol
  • water 2 mL
  • LiOH 9 mg, 0.38 mmol
  • Step 2 methyl 8-bromo-2-formylimidazo[1,2-a]pyridine-6-carboxylate [0185] To a suspension of methyl 8-bromo-2-(dichloromethyl)imidazo[1,2-a]pyridine-6- carboxylate (2 g, 5.92 mmol) in MeOH (30 mL) and water (10 mL) was added NaOAc (1.456 g, 17.75 mmol) then the mixture was heated to 80 °C. After 12 h the mixture was cooled to rt then extracted with DCM (50 mL x 2). The combined organic extracts were washed with brine (30 mL x 2) then dried (Na2SO4) then filtered then the filtrate was concentrated.
  • Step 3 methyl 8-bromo-2-(2,2,2-trifluoro-1-hydroxyethyl)imidazo[1,2-a]pyridine-6- carboxylate
  • Trimethyl(trifluoromethyl)silane 1.055 g, 7.42 mmol was added to a degassed solution of methyl 8-bromo-2-formylimidazo[1,2-a]pyridine-6-carboxylate (1.05 g, 3.71 mmol) in DME (5 mL) at 0 °C.
  • Step 4 methyl 8-bromo-2-(2,2,2-trifluoroethyl)imidazo[1,2-a]pyridine-6-carboxylate
  • Step 5 methyl 8-methoxy-2-(2,2,2-trifluoroethyl)imidazo[1,2-a]pyridine-6-carboxylate 25700 [0188] Methyl 8-bromo-2-(2,2,2-trifluoroethyl)imidazo[1,2-a]pyridine-6-carboxylate (150 mg, 0.34 mmol), Cs2CO3 (221 mg, 0.68 mmol), and methanol (109 mg, 3.40 mmol) were combined in toluene (10 mL) in a glove box.
  • Step 6 8-methoxy-2-(2,2,2-trifluoroethyl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • a solution of methyl 8-methoxy-2-(2,2,2-trifluoroethyl)imidazo[1,2-a]pyridine-6- carboxylate (60 mg, 0.21 mmol) in THF (2 mL) and water (2 mL) was added NaOH (24.98 mg, 0.625 mmol) at rt. After 4 h the mixture was concentrated to remove volatile organic solvents. The pH of the resulting mixture was adjusted to 5 using 1M HCl then it was extracted with EtOAc (10 mL x 3).
  • Step 2 methyl 8-acetyl-2-(difluoromethyl)imidazo[1,2-a]pyridine-6-carboxylate [0191] To a solution of methyl 8-bromo-2-(difluoromethyl)imidazo[1,2-a]pyridine-6- carboxylate (240 mg, 0.79 mmol) in 1,4-dioxane (5 mL) was added tributyl(1-ethoxyvinyl)tin (0.2 mL, 0.56 mmol) and Pd(Ph 3 P) 4 (91 mg, 0.079 mmol) then the mixture was heated to 110 °C under N2.
  • Step 3 methyl 2-(difluoromethyl)-8-(1-hydroxyethyl)imidazo[1,2-a]pyridine-6-carboxylate [0192] To a mixture of methyl 8-acetyl-2-(difluoromethyl)imidazo[1,2-a]pyridine-6- carboxylate (116 mg, 0.432 mmol) in MeOH (10 mL) was added NaBH 4 (8.2 mg, 0.216 mmol) at 0 °C. After 10 minutes the mixture was quenched with water (10 mL) then warmed to rt then extracted with EtOAc (20 mL x 3).
  • Step 4 2-(difluoromethyl)-8-(1-hydroxyethyl)imidazo[1,2-a]pyridine-6-carboxylic acid [0193] To a mixture of methyl 2-(difluoromethyl)-8-(1-hydroxyethyl)imidazo[1,2-a]pyridine-6- carboxylate (60 mg, 0.222 mmol) in THF (10 mL) was added LiOH (53.2 mg, 2.22 mmol) in water (2 mL) at rt.
  • Step 3 6-bromo-2-(difluoromethyl)-8-iodoimidazo[1,2-a]pyridine [0196] To a solution of 6-bromo-8-iodoimidazo[1,2-a]pyridine-2-carbaldehyde (350 mg, 0.997 mmol) in DCM (10 ml) was added DAST (0.659 ml, 4.99 mmol) at -78 °C. After the addition was complete the mixture was warmed to rt. After 12 h the mixture was diluted with saturated NaHCO3 (5 mL) then extracted with DCM (10 mL x 3).
  • Step 4 6-bromo-8-cyclopropyl-2-(difluoromethyl)imidazo[1,2-a]pyridine
  • a glove box To a solution of 6-bromo-2-(difluoromethyl)-8-iodoimidazo[1,2- a]pyridine (100 mg, 0.268 mmol) and cyclopropylboronic acid (69.1 mg, 0.804 mmol) in toluene (2 mL) was added Pd(OAc) 2 (6.0 mg, 0.027 mmol), Na 2 CO 3 (85 mg, 0.804 mmol) and tricyclohexylphosphene (15.0 mg, 0.054 mmol) then the mixture was heated to 100 °C.
  • Step 2 methyl 8-acetyl-2-methylimidazo[1,2-a]pyridine-6-carboxylate [0200] To a solution of methyl 8-bromo-2-methylimidazo[1,2-a]pyridine-6-carboxylate (1.5 g, 5.57 mmol) in 1,4-dioxane (30 mL) were added tributyl(1-ethoxyvinyl)stannane (4.9 mL, 14.62 mmol) and Pd(Ph3P)4 (664 mg, 0.557 mmol) then the mixture was heated to 110 °C. After 12 h the mixture was cooled to rt then 4M HCl (20 mL) was added.
  • Step 3 methyl 8-(1,1-difluoroethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate [0201] To a mixture of methyl 8-acetyl-2-methylimidazo[1,2-a]pyridine-6-carboxylate (500 mg, 2.153 mmol) in DCM (10 mL) was added DAST (8 mL, 60.6 mmol) at 0 °C then the mixture was warmed to rt. After 18 h the mixture was diluted with saturated NaHCO3 (15 mL) then extracted with EtOAc (15 mL x 3).
  • Step 4 8-(1,1-difluoroethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • a mixture of methyl 8-(1,1-difluoroethyl)-2-methylimidazo[1,2-a]pyridine-6- carboxylate (150 mg, 0.59 mmol) in THF (5 mL) was added LiOH (28.3 mg, 1.18 mmol) in water (0.5 mL) at rt. After 1 h the mixture was diluted with H2O (15 mL) then the pH adjusted to 3 with 1M HCl. The resulting mixture was extracted with EtOAc (10 mL x 4).
  • Step 2 methyl 8-(1-methoxyethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate
  • Sodium hydride (20.5 mg, 0.51 mmol, 60% dispersion in mineral oil) was added to a solution of methyl 8-(1-hydroxyethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate (40 mg, 0.17 mmol) in DMF (2.5 mL) at 0 °C.
  • DMF dimethyl 8-(1-hydroxyethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate
  • Step 3 8-(1-methoxyethyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • a mixture of methyl 8-(1-methoxyethyl)-2-methylimidazo[1,2-a]pyridine-6- carboxylate (21 mg, 0.068 mmol) in THF (2 mL) and water (2 mL) was added lithium hydroxide hydrate (14.2 mg, 0.34 mmol) at rt. After 2 h the mixture was concentrated to remove volatile organic solvents. The resulting aqueous mixture was washed with DCM (5 mL x 2) then the pH was adjusted to 6 using 1M HCl then the mixture was concentrated.
  • TBDMSOTf (683 mg, 2.58 mmol) dropwise. The mixture was slowly warmed to rt over 3 h. The resulting mixture was diluted with DCM (30 mL) then washed with saturated NaHCO 3 (20 mL), water, and brine (20 mL). The organic layer was dried (Na2SO4) then filtered then the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-30% EtOAc/petroleum ether) to give the title compound.
  • Step 2 methyl 8-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)-2-methylimidazo[1,2- a]pyridine-6-carboxylate
  • diethylzinc (1M in toluene, 4.8 mL, 4.81 mmol) in DCM (20 mL) at - 4°C was added a solution of chloroiodomethane (1.7 g, 9.62 mmol) in DCM (5 mL) dropwise. Once the addition was complete the mixture was warmed to 0 °C.
  • Step 3 methyl 8-(1-hydroxycyclopropyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate
  • TBAF TBAF
  • Step 4 8-(1-hydroxycyclopropyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • Step 2 8-(1-fluorocyclopropyl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • Step 2 methyl 8-(isoxazol-3-yl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate
  • Step 3 8-(isoxazol-3-yl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • a mixture of methyl 8-(isoxazol-3-yl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate (42 mg, 0.16 mmol) in THF (1 mL) and water (0.6 mL) was added LiOH (20 mg, 0.82 mmol) at rt. After 2 h the mixture was concentrated to remove volatile organic solvents.
  • Step 2 methyl (Z)-8-(N'-hydroxycarbamimidoyl)-2-methylimidazo[1,2-a]pyridine-6- carboxylate [0217] To a solution of methyl 8-cyano-2-methylimidazo[1,2-a]pyridine-6-carboxylate (1.1 g, 5.11 mmol) in EtOH (50 mL) was added NaHCO3 (1.116 g, 13.29 mmol) and hydroxylamine hydrochloride (0.910 g, 13.29 mmol) then the mixture was heated to 80 °C. After 8 h the mixture was cooled to rt then filtered then the filtrate was concentrated to give the title compound which was used without purification.
  • Step 3 methyl 2-methyl-8-(1,2,4-oxadiazol-3-yl)imidazo[1,2-a]pyridine-6-carboxylate [0218] To a solution of methyl (Z)-8-(N'-hydroxycarbamimidoyl)-2-methylimidazo[1,2- a]pyridine-6-carboxylate (1.5 g, 6.04 mmol) in trimethyl orthoformate (50 g, 471 mmol) was added pTsOH (0.104 g, 0.604 mmol) then the mixture was heated to 100 °C. After 12 h the mixture was cooled to rt then concentrated.
  • Step 2 8-(benzyloxy)-6-bromo-2-methylimidazo[1,2-a]pyridine [0222] To a solution of 3-(benzyloxy)-5-bromopyridin-2-amine (2 g, 7.16 mmol) in EtOH (40 mL) was added 1-bromopropan-2-one (2.1 mL, 25.6 mmol) then the mixture was heated to 80 °C.
  • Step 3 ethyl 8-(benzyloxy)-2-methylimidazo[1,2-a]pyridine-6-carboxylate
  • Pd(dppf)Cl2 268 mg, 0.37 mmol
  • Step 4 ethyl 8-hydroxy-2-methylimidazo[1,2-a]pyridine-6-carboxylate
  • ethyl 8-(benzyloxy)-2-methylimidazo[1,2-a]pyridine-6-carboxylate 680 mg, 2.19 mmol
  • MeOH MeOH
  • Pd/C 466 mg, 0.44 mmol
  • Step 5 8-hydroxy-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid 25700 [0225] To a solution of ethyl 8-hydroxy-2-methylimidazo[1,2-a]pyridine-6-carboxylate (200 mg, 0.91 mmol) in THF (2 mL) and water (2 mL) was added NaOH (109 mg, 4.54 mmol) at rt. After 2 h the mixture was concentrated to remove volatile organic solvents. The pH of the resulting mixture was adjusted to 5 using 1M HCl then it was extracted with EtOAc (10 mL x 3). The combined organic extracts were dried (Na2SO4) then filtered then the filtrate was concentrated to give the title compound.
  • Step 2 methyl rac-2-(2,2-difluorocyclopropyl)imidazo[1,2-a]pyridine-6-carboxylate
  • methyl 2-vinylimidazo[1,2-a]pyridine-6-carboxylate 400 mg, 1.98 mmol
  • toluene 15 mL
  • TMSCF2Br 0.9 mL, 5.93 mmol
  • TBAB TBAB (19.1 mg, 0.059 mmol
  • Step 2 5-(5-bromo-2-chloropyridin-3-yl)-1,3,4-oxathiazol-2-one 25700 [0230] To a solution of the 5-bromo-2-chloronicotinamide (3 g, 12.74 mmol) in THF (80 mL) was added carbonochloridic hypochlorous thioanhydride (3.34 g, 25.5 mmol) at rt. After 16 h the mixture was concentrated.
  • Step 3 3-(5-bromo-2-chloropyridin-3-yl)isothiazole [0231] A mixture of 5-(5-bromo-2-chloropyridin-3-yl)-1,3,4-oxathiazol-2-one (2.2 g, 7.50 mmol) and bicyclo[2.2.1]hepta-2,5-diene (6.91 g, 75.0 mmol) in xylene (20 mL) was heated under microwave conditions to 170 °C for 1 h. The resulting mixture was concentrated.
  • Step 4 ethyl 6-chloro-5-(isothiazol-3-yl)nicotinate
  • 3-(5-bromo-2-chloropyridin-3-yl)isothiazole 800 mg, 2.90 mmol
  • isopropylmagnesium chloride-lithium chloride complex 1.3M in hexanes, 2.68 mL, 3.48 mmol
  • ethyl chloroformate 1.1 mL, 11.61 mmol
  • Step 5 ethyl 6-amino-5-(isothiazol-3-yl)nicotinate [0233] To a solution of ethyl 6-chloro-5-(isothiazol-3-yl)nicotinate (450 mg, 1.68 mmol) in 1,4- dioxane (15 mL) was added Cs2CO3 (1310 mg, 4.02 mmol), Xantphos (194 mg, 0.34 mmol), diphenylmethanimine (364 mg, 2.01 mmol) and Pd2(dba)3 (153 mg, 0.17 mmol) under N2 then the mixture was heated to 90 °C. After 2 h the mixture was cooled to rt then concentrated.
  • Step 6 ethyl 8-(isothiazol-3-yl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate [0234]
  • a mixture of 1-bromopropan-2-one (0.3 mL, 3.61 mmol) and ethyl 6-amino-5- (isothiazol-3-yl)nicotinate (90 mg, 0.361 mmol) in EtOH (6 mL) was heated to 80 °C. After 12 h the mixture was cooled to rt then diluted with water (10 mL) then extracted with EtOAc (10 mL x 3). The combined organic extracts were dried (Na2SO4) then filtered then the filtrate was 25700 concentrated.
  • Step 7 8-(isothiazol-3-yl)-2-methylimidazo[1,2-a]pyridine-6-carboxylic acid
  • ethyl 8-(isothiazol-3-yl)-2-methylimidazo[1,2-a]pyridine-6-carboxylate 95 mg, 0.33 mmol
  • water 2 mL
  • lithium hydroxide 23.8 mg, 0.99 mmol
  • Step 2 methyl 2-cyclopropyl-8-methoxyimidazo[1,2-a]pyrazine-6-carboxylate
  • Step 2 methyl 2-cyclopropyl-8-methoxyimidazo[1,2-a]pyrazine-6-carboxylate
  • Step 3 2-cyclopropyl-8-methoxyimidazo[1,2-a]pyrazine-6-carboxylic acid
  • MeOH methyl 2-cyclopropyl-8-methoxyimidazo[1,2-a]pyrazine-6-carboxylate
  • 6M NaOH 0.539 mL, 3.24 mmol
  • Step 2 methyl 8-bromo-2-methyl-[1,2,4]triazolo[1,5-a]pyridine-6-carboxylate [0241] To a solution of 1,2-diamino-3-bromo-5-(methoxycarbonyl)pyridin-1-ium 2,4,6- trimethylbenzenesulfonate (8.0 g, 17.92 mmol) in pyridine (24 mL) was added acetyl chloride (7.04 g, 90 mmol) then the mixture was heated to 100 oC. After 1 h the mixture was cooled to rt then concentrated.
  • Step 3 methyl 8-methoxy-2-methyl-[1,2,4]triazolo[1,5-a]pyridine-6-carboxylate
  • Step 2 diethyl 2-methylpyrazolo[1,5-a]pyridine-3,5-dicarboxylate [0246] A mixture of 1-amino-4-(ethoxycarbonyl)pyridin-1-ium 2,4-dinitrophenolate (26 g, 74.2 mmol) and K2CO3 (10.26 g, 74.2 mmol) in EtOH (70 mL) was stirred at rt for 30 minutes then ethyl 3-oxobutanoate (9.66 mL, 74.2 mmol) was added. The resulting mixture was heated to 60 °C. After 5 h the mixture was cooled to rt then diluted with H2O then the mixture was extracted with EtOAc (3 x 50 mL).
  • Step 3 diethyl 7-bromo-2-methylpyrazolo[1,5-a]pyridine-3,5-dicarboxylate [0247] A solution of diethyl 2-methylpyrazolo[1,5-a]pyridine-3,5-dicarboxylate (4 g, 14.48 mmol) in THF (100 mL) was cooled to -78 °C.
  • Step 4 7-methoxy-3-(methoxycarbonyl)-2-methylpyrazolo[1,5-a]pyridine-5-carboxylic acid
  • Diethyl 7-bromo-2-methylpyrazolo[1,5-a]pyridine-3,5-dicarboxylate 2.5 g, 7.00 mmol
  • MeOH 30 mL
  • NaOMe 0.702 g, 12.99 mmol
  • Step 5 7-methoxy-2-methylpyrazolo[1,5-a]pyridine-3,5-dicarboxylic acid
  • Step 2 1-amino-2-bromo-6-(cyclopropylethynyl)-4-(methoxycarbonyl)pyridin-1-ium 2,4,6- trimethylbenzenesulfonate 25700 [0253] A mixture of TFA (22.2 mL) and water (2 mL) was cooled to 0 °C then ethyl O- mesitylsulfonylacetohydroxamate (8.21 g, 28.8 mmol) was added. After 1.5 h ice water (100 mL) was added. The resulting precipitate was collected by filtration then the cake was washed with water (10 x 10 mL).
  • Step 3 methyl 7-bromo-2-cyclopropylpyrazolo[1,5-a]pyridine-5-carboxylate
  • Step 4 methyl 2-cyclopropylpyrazolo[1,5-a]pyridine-5-carboxylate and methyl 2- propylpyrazolo[1,5-a]pyridine-5-carboxylate [0255] Methyl 7-bromo-2-cyclopropylpyrazolo[1,5-a]pyridine-5-carboxylate (150 mg, 0.51 mmol), NaOAc (41.7 mg, 0.51 mmol), and 10% Pd/C (5.4 mg, 0.051 mmol) were combined under N2.
  • Step 5 2-cyclopropylpyrazolo[1,5-a]pyridine-5-carboxylic acid and 2-propylpyrazolo[1,5- a]pyridine-5-carboxylic acid
  • a solution of a mixture of methyl 2-cyclopropylpyrazolo[1,5-a]pyridine-5- carboxylate and methyl 2-propylpyrazolo[1,5-a]pyridine-5-carboxylate 68 mg of a 3:1 mixture of c-Pr to n-Pr, 0.314 mmol
  • MeOH 3.15 mL
  • Step 2 7-bromo-2-methylpyrazolo[1,5-a]pyridine-5-carboxylic acid
  • Step 4 methyl 2-methyl-7-(trifluoromethyl)pyrazolo[1,5-a]pyridine-5-carboxylate [0261] To a mixture of methyl 7-bromo-2-methylpyrazolo[1,5-a]pyridine-5-carboxylate (300 mg, 1.12 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (643 mg, 3.34 mmol) in DMF (10 mL) was added copper (I) iodide (425 mg, 2.23 mmol) under N 2 then the mixture was heated to 80 °C. After 18 h the mixture was cooled to rt then diluted with saturated NaHCO3 (30 mL).
  • Step 2 methyl 7-(1-hydroxyethyl)-2-methylpyrazolo[1,5-a]pyridine-5-carboxylate [0264] To a mixture of methyl 7-acetyl-2-methylpyrazolo[1,5-a]pyridine-5-carboxylate (80 mg, 0.34 mmol) in MeOH (3 mL) was added NaBH4 (13 mg, 0.34 mmol) at 0 °C. After 30 minutes the mixture was quenched with H 2 O (10 mL) then warmed to rt then extracted with EtOAc (5 mL x 3). The combined organic phases were washed with brine (10 mL) then dried (Na2SO4) then filtered then the filtrate was concentrated.
  • Step 3 7-(1-hydroxyethyl)-2-methylpyrazolo[1,5-a]pyridine-5-carboxylic acid
  • Step 2 methyl 7-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)-2-methylpyrazolo[1,5- a]pyridine-5-carboxylate [0267] To a solution of diethylzinc (1M in toluene, 0.6 mL 0.58 mmol) in DCM (1 mL) was added a solution of chloroiodomethane (204 mg, 1.16 mmol) in DCM (0.5 mL) dropwise at -4 °C.
  • Step 3 methyl 7-(1-hydroxycyclopropyl)-2-methylpyrazolo[1,5-a]pyridine-5-carboxylate 25700 [0268] To a solution of methyl 7-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)-2- methylpyrazolo[1,5-a]pyridine-5-carboxylate (45 mg, 0.125 mmol) in THF (2 mL) was added TBAF (1M in THF, 0.125 mL 0.125 mmol) at rt. After 1 h the mixture was diluted with water (5 mL) then extracted with EtOAc (5 mL x 3).
  • Step 4 7-(1-hydroxycyclopropyl)-2-methylpyrazolo[1,5-a]pyridine-5-carboxylic acid
  • Step 2 2-methylpyrazolo[1,5-a]pyridine-5-carboxylic acid
  • ethyl 2-methylpyrazolo[1,5-a]pyridine-5-carboxylate 150 mg, 0.73 mmol
  • EtOH 3 mL
  • water 1.5 mL
  • Step 2 methyl (E)-7-((hydroxyimino)methyl)-2-methylpyrazolo[1,5-a]pyridine-5- carboxylate [0273] To a solution of methyl 7-formyl-2-methylpyrazolo[1,5-a]pyridine-5-carboxylate (150 mg, 0.55 mmol) in EtOH (4 mL) was added NaHCO3 (139 mg, 1.65 mmol) and hydroxylamine hydrochloride (76 mg, 1.10 mmol) at rt. After 2 h the mixture was concentrated to give the title compound which was used in the next step without purification.
  • Step 3 methyl 2-methyl-7-(5-(trimethylsilyl)isoxazol-3-yl)pyrazolo[1,5-a]pyridine-5- carboxylate 25700 [0274] A solution of methyl (E)-7-((hydroxyimino)methyl)-2-methylpyrazolo[1,5-a]pyridine-5- carboxylate (100 mg, 0.34 mmol) and trimethylsilylacetylene (0.153 ml, 1.03 mmol) in MeCN (2 mL) was added to CrO 2 (288 mg, 3.43 mmol) then the mixture was heated to 80 °C. After 12 h the mixture was cooled to rt then filtered then the filtrate was concentrated.
  • Step 4 7-(isoxazol-3-yl)-2-methylpyrazolo[1,5-a]pyridine-5-carboxylic acid
  • Step 2 methyl 3,5-dibromo-4-ethanethioamidobenzoate
  • a solution of methyl 4-acetamido-3,5-dibromobenzoate (240 mg, 0.68 mmol) and Lawesson's reagent (138 mg, 0.34 mmol) in toluene (5 mL) was heated to 110 °C. After 2 h the 25700 mixture was cooled to rt then concentrated. The crude product was subjected to silica gel chromatography (0-30% EtOAc/petroleum ether) to give the title compound.
  • Step 3 methyl 4-bromo-2-methylbenzo[d]thiazole-6-carboxylate [0278] To a solution of methyl 3,5-dibromo-4-ethanethioamidobenzoate (100 mg, 0.27 mmol) in DME (5 mL) was added CuI (3 mg, 0.014 mmol), 1,10-phenanthroline (5 mg, 0.027 mmol) and Cs 2 CO 3 (133 mg, 0.41 mmol). The resulting mixture was heated to 85° C. After 20 h the mixture was cooled to rt then filtered through a pad of Celite ® . The filtrate was concentrated.
  • Step 4 4-bromo-2-methylbenzo[d]thiazole-6-carboxylic acid
  • a mixture of methyl 4-bromo-2-methylbenzo[d]thiazole-6-carboxylate (63 mg, 0.22 mmol) in THF (3 mL) H 2 O (1.8 mL) and EtOH (3 mL) was added NaOH (44.0 mg, 1.10 mmol) at rt. After 2 h the mixture was concentrated to remove volatile organic solvents.
  • Step 2 ethyl 2-bromo-4-(trifluoromethyl)benzo[d]thiazole-6-carboxylate
  • ethyl 2-amino-4-(trifluoromethyl)benzo[d]thiazole-6-carboxylate 2.5 g, 6.89 mmol
  • MeCN 70 ml
  • copper (I) bromide 1.83 g, 10.34 mmol
  • tert- butyl nitrite (2.132 g, 20.67 mmol
  • Step 3 ethyl 2-methyl-4-(trifluoromethyl)benzo[d]thiazole-6-carboxylate
  • methylboronic acid 50.7 mg, 0.85 mmol
  • K 3 PO 4 240 mg, 1.13 mmol
  • 1,4-dioxane 5 mL
  • Pd(dppf)Cl2 31.0 mg, 0.042 mmol
  • Step 4 2-methyl-4-(trifluoromethyl)benzo[d]thiazole-6-carboxylic acid [0283] To a solution of ethyl 2-methyl-4-(trifluoromethyl)benzo[d]thiazole-6-carboxylate (60 mg, 0.21 mmol) in THF (2 mL) and water (2 mL) was added NaOH (24.8 mg, 0.62 mmol) at rt. After 12 h the mixture was concentrated to remove volatile organic solvents.
  • Step 2 methyl 2-methyl-4-vinylbenzo[d]oxazole-6-carboxylate [0285] To a mixture of methyl 4-bromo-2-methylbenzo[d]oxazole-6-carboxylate (1 g, 3.70 mmol), K2CO3 (0.768 g, 5.55 mmol), and potassium vinyltrifluoroborate (0.595 g, 4.44 mmol) in water (1.5 mL) and 1,4-dioxane (15 mL) was added Pd(dppf)Cl 2 (0.542 g, 0.741 mmol) under N 2 then the mixture was heated to 100 °C.
  • Step 3 methyl 4-formyl-2-methylbenzo[d]oxazole-6-carboxylate [0286] To a solution of methyl 2-methyl-4-vinylbenzo[d]oxazole-6-carboxylate (580 mg, 2.67 mmol) and NMO (626 mg, 5.34 mmol) in THF (10 mL) was added K2OsO4 ⁇ 2H2O (197 mg, 0.534 mmol) at rt. After 1 h NaIO 4 (1.71 g, 8.01 mmol) was added. After 12 h the mixture was diluted with saturated Na2SO3 (30 mL) then extracted with EtOAc (30 mL x 3).
  • Step 4 methyl (E)-4-((hydroxyimino)methyl)-2-methylbenzo[d]oxazole-6-carboxylate [0287] To a solution of NaHCO3 (460 mg, 5.47 mmol) and hydroxylamine hydrochloride (190 mg, 2.74 mmol) in EtOH (10 mL) was added methyl 4-formyl-2-methylbenzo[d]oxazole-6- carboxylate (400 mg, 1.83 mmol) at rt. After 3 h the mixture was concentrated to give the title compound which was used in the next step without purification.
  • Step 5 methyl 2-methyl-4-(5-(trimethylsilyl)isoxazol-3-yl)benzo[d]oxazole-6-carboxylate [0288] To a solution of methyl (E)-4-((hydroxyimino)methyl)-2-methylbenzo[d]oxazole-6- carboxylate (400 mg, 1.71 mmol) and trimethylsilylacetylene (503 mg, 5.12 mmol) in MeCN (20 mL) was added CrO 2 (1.435 g, 17.08 mmol) then the mixture was heated to 80 °C. After 12 h the mixture was cooled to rt then filtered through a thin pad of silica gel washing with EtOAc.
  • Step 6 4-(isoxazol-3-yl)-2-methylbenzo[d]oxazole-6-carboxylic acid
  • methyl 2-methyl-4-(5-(trimethylsilyl)isoxazol-3-yl)benzo[d]oxazole-6- carboxylate 120 mg, 0.36 mmol
  • water 5 mL
  • LiOH 43.5 mg, 1.82 mmol
  • Step 2 ethyl 6-chloro-5-(2-oxo-1,3,4-oxathiazol-5-yl)nicotinate
  • ethyl 5-carbamoyl-6-chloronicotinate (1 g, 4.37 mmol) in THF (30 mL) was added carbonochloridic hypochlorous thioanhydride (1.72 g, 13.12 mmol) at rt. After 16 h the mixture was concentrated. The crude product was subjected to silica gel chromatography (0- 60% EtOAc/petroleum ether) to give the title compound.
  • Step 3 ethyl 3-(2-chloro-5-(ethoxycarbonyl)pyridin-3-yl)-1,2,4-thiadiazole-5-carboxylate
  • a mixture of ethyl 6-chloro-5-(2-oxo-1,3,4-oxathiazol-5-yl)nicotinate (300 mg, 1.046 mmol) and ethyl carbonocyanidate (1037 mg, 10.46 mmol) in xylene (6 mL) was sealed in a 40 mL microwave vial then heated to 170 °C by microwave irradition for 1 h. The resulting mixture was cooled to rt then concentrated.
  • Step 5 ethyl 3-(6-(ethoxycarbonyl)-2-methylimidazo[1,2-a]pyridin-8-yl)-1,2,4-thiadiazole-5- carboxylate
  • a mixture of 1-bromopropan-2-one (0.5 mL, 5.58 mmol) and ethyl 3-(2-amino-5- (ethoxycarbonyl)pyridin-3-yl)-1,2,4-thiadiazole-5-carboxylate (180 mg, 0.558 mmol) in EtOH (6 mL) was heated to 80 °C. After 12 h the mixture was cooled to rt then diluted with H2O (10 mL).
  • Step 6 2-methyl-8-(1,2,4-thiadiazol-3-yl)imidazo[1,2-a]pyridine-6-carboxylic acid
  • Step 2 methyl 3-cyclopropoxy-4-nitro-5-vinylbenzoate
  • Step 3 methyl 3-cyclopropoxy-5-formyl-4-nitrobenzoate
  • a solution of methyl 3-cyclopropoxy-4-nitro-5-vinylbenzoate (0.7 g, 2.66 mmol) in DCM (26 mL) was cooled to -78 o C then treated with O3 (excess) until TLC indicated starting material had been consumed. Excess O 3 was removed by purging with O 2 then N 2 . The reaction was quenched with Ph3P (2.092 g, 7.98 mmol) then warmed to 25 o C. After 1 h the mixture was concentrated.
  • Step 4 methyl 3-cyclopropoxy-5-((methylamino)methyl)-4-nitrobenzoate
  • MeOH a solution of methyl 3-cyclopropoxy-5-formyl-4-nitrobenzoate (450 mg, 1.697 mmol) in MeOH (34 mL) was added methylamine as an EtOH solution (527 mg, 5.09 mmol) and AcOH (0.583 mL, 10.18 mmol) at 0 o C.
  • NaBH(OAc)3 (2877 mg, 13.57 mmol) was added then the mixture was warmed to rt.
  • Step 5 methyl 7-cyclopropoxy-2-methyl-2H-indazole-5-carboxylate [0300] To a solution of methyl 3-cyclopropoxy-5-((methylamino)methyl)-4-nitrobenzoate (450 mg, 1.606 mmol) in MeOH (30 mL) was added zinc (840 mg, 12.84 mmol) then dropwise addition of ammonium formate (202 mg, 3.21 mmol) in MeOH (1.5 mL) at rt. After 3 h the mixture was filtered through a pad of Celite ® washing with MeOH then the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-50% EtOAc/petroleum ether) to give the title compound.
  • Step 6 7-cyclopropoxy-2-methyl-2H-indazole-5-carboxylic acid [0301] To a solution of methyl 7-cyclopropoxy-2-methyl-2H-indazole-5-carboxylate (200 mg, 0.812 mmol) in THF (3 mL) and MeOH (3 mL) was added lithium hydroxide monohydrate (85 mg, 2.030 mmol) in water (1 mL) at ambient temperature. After 2 h the mixture was poured into ice-water (25 mL) then washed with EtOAc (10 mL). The aqueous phase was acidified to pH 2 using 1N HCl then the resulting mixture was extracted with EtOAc (3 x 15 mL).
  • Step 2 6-bromo-8-(difluoromethoxy)imidazo[1,2-a]pyridine-2-carbaldehyde
  • DCM dimethylethyl sulfoxide
  • Step 2 2-(6-bromo-2-methylimidazo[1,2-a]pyridin-8-yl)propan-2-ol 25700 [0310] To a solution of methyl 6-bromo-2-methylimidazo[1,2-a]pyridine-8-carboxylate (200 mg, 0.74 mmol) in THF (10 mL) was added methylmagnesium bromide (3M in THF, 1.5 mL, 4.50 mmol) slowly at 0 °C. After 1 h the mixture was quenched with saturated NH 4 Cl (10 mL) then warmed to rt then extracted with EtOAc (40 mL x 3).
  • Step 2 rac-(6-bromo-2-methylimidazo[1,2-a]pyridin-8-yl)(cyclopropyl)methanol
  • 6-bromo-2-methylimidazo[1,2-a]pyridine-8-carbaldehyde 100 mg, 0.42 mmol
  • cyclopropylmagnesium bromide 0.5M in THF, 2.5 mL, 1.26 mmol
  • Step 2 2-(6-bromo-2-methylimidazo[1,2-a]pyridin-8-yl)-1,3,4-oxadiazole
  • Step 2 6-bromo-2-methylimidazo[1,2-a]pyridine-8-carboxamide
  • HATU 596 mg, 1.57 mmol
  • DIPEA 0.4 mL, 2.35 mmol
  • NH4Cl 84 mg, 1.57 mmol
  • Step 3 2-(6-bromo-2-methylimidazo[1,2-a]pyridin-8-yl)-4-methyloxazole [0317] A mixture of 1-bromopropan-2-one (1.21 g, 8.83 mmol), AgOTf (0.364 g, 1.42 mmol), and 6-bromo-2-methylimidazo[1,2-a]pyridine-8-carboxamide (180 mg, 0.71 mmol) in EtOAc (10 mL) was heated to 80 °C.
  • Step 2 3-cyclopropoxypyridin-2-amine [0319] A mixture of 3-cyclopropoxy-2-nitropyridine (1 g, 5.55 mmol), iron (3.10 g, 55.5 mmol), and ammonia hydrochloride (2.97 g, 55.5 mmol) in EtOH (10 mL) and AcOH (1mL) was heated to 80 °C. After 12 h the mixture was cooled to rt then filtered then the filtrate was concentrated.
  • Step 3 5-bromo-3-cyclopropoxypyridin-2-amine [0320] To a solution of 3-cyclopropoxypyridin-2-amine (510 mg, 3.40 mmol) in AcOH (10 mL) was added Br2 (0.4 mL, 6.79 mmol) at rt. After 12 h the mixture was concentrated.
  • Step 4 6-bromo-8-cyclopropoxy-2-methylimidazo[1,2-a]pyridine [0321] To a solution of 5-bromo-3-cyclopropoxypyridin-2-amine (320 mg, 1.40 mmol) in EtOH (5 mL) was added 1-bromopropan-2-one (0.2 mL, 2.79 mmol) then the mixture was heated to 80 °C. After 12 h the mixture was cooled to rt then concentrated. The residue was taken up in saturated NaHCO3 then the mixture was extracted with DCM (10 mL x 2). The combined organic extracts were washed with brine (20 mL x 2) then dried (Na 2 SO 4 ) then filtered then the filtrate was concentrated.
  • Step 3 1-(6-bromoimidazo[1,2-a]pyridin-2-yl)ethan-1-ol [0325] To a solution of 6-bromoimidazo[1,2-a]pyridine-2-carbaldehyde (400 mg, 1.78 mmol) in THF (5 mL) was added methylmagnesium bromide (3M in Et 2 O, 2.96 mL, 8.89 mmol) slowly at -78 °C.
  • Step 2 6-bromo-2-(1,1-difluoroethyl)imidazo[1,2-a]pyridine [0327] To a solution of 1-(6-bromoimidazo[1,2-a]pyridin-2-yl)ethan-1-one (250 mg, 1.05 mmol) in DCM (2 mL) was added DAST (1.4 mL, 10.46 mmol) at -78 °C. The mixture was allowed to warm to rt slowly over 12 h. The resulting mixture was diluted with saturated NaHCO3 (20 mL) then the mixture was extracted with DCM (20 mL x 2).
  • Step 2 6-bromo-2-(1-fluorocyclopropyl)imidazo[1,2-a]pyridine [0329] To a solution of 1-(6-bromoimidazo[1,2-a]pyridin-2-yl)cyclopropan-1-ol (55 mg, 0.22 mmol) in DCM (5 mL) was added DAST (0.2 mL, 1.09 mmol) at -78 °C.
  • Step 2 6-bromo-8-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carbaldehyde
  • DCM dimethyl sulfoxide
  • Step 3 (6-bromo-8-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methanol [0332] To a solution of 6-bromo-8-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carbaldehyde (150 mg, 0.52 mmol) in EtOH (6 ml) was added NaBH4 (19.36 mg, 0.52 mmol) at 0 °C.
  • Step 4 6-bromo-2-(fluoromethyl)-8-(trifluoromethyl)imidazo[1,2-a]pyridine
  • DCM dimethylethyl sulfoxide
  • DAST 0.4 mL, 2.54 mmol
  • the mixture was allowed to warm to rt slowly over 12 h.
  • the resulting mixture was diluted with saturated NaHCO 3 (5 mL) then extracted with DCM (10 mL x 3).
  • Triethylamine (1.0 mL, 7.4 mmol) was added then the mixture was heated to 45 °C for 4 h. The resulting mixture was cooled to rt then concentrated. The crude product was subjected to silica gel chromatography (0-15% EtOAc/hexanes) to give the title compound.
  • Step 3 7,7-difluoro-3-iodobicyclo[4.2.0]octa-1(6),2,4-triene
  • a mixture of bis(2-methoxyethyl)aminosulfur trifluoride (0.93 mL, 5.0 mmol) and 3- iodobicyclo[4.2.0]octa-1(6),2,4-trien-7-one (250 mg, 1.0 mmol) was stirred at 50 °C for 16 h. The mixture was cooled to rt and a 0 °C solution of NaHCO3 (saturated aq., 30 mL) was added dropwise.
  • Step 2 ((1R,5S,6s)-6-hydroxy-3-azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8- (trifluoromethyl)imidazo[1,2-a]pyridin-6-yl)methanone [0338] To a solution of 2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carboxylic acid (Int.
  • Step 2 ent-((1R,5S,6s)-6-((4-(2-amino-1-fluoropropan-2-yl)-6-(4-fluorophenyl)pyridin-2- yl)oxy)-3-azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone (enantiomer 1 and 2) [0345] To a solution of rac-benzyl (1-fluoro-2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl- 8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carbonyl)
  • Example 1-1-P1 (faster eluting enantiomer): MS m/z (M+H) + : calculated 572.2, observed 572.1.
  • Example 2-1-P1 ent-((1R,5S,6s)-6-((6-(4-fluorophenyl)-4-(2-methylpyrrolidin-2-yl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone (enantiomer 1) and Example 2-1-P2 ent-((1R,5S,6s)-6-((6-(4-fluorophenyl)-4-(2-methylpyrrolidin-2-yl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone (enantiomer 2) Step 1: ent-tert-butyl 8- (trifluoro
  • the racemate was subjected to chiral SFC (ChiralPak AS, 30% EtOH (with 0.1% NH4OH modifier)/CO 2 ) to give the title compounds.
  • Example Int.2-1-P1 faster eluting enantiomer
  • Example Int.2-1-P2 lower eluting enantiomer
  • Step 2 ent-((1R,5S,6s)-6-((4-(2-amino-1-fluoropropan-2-yl)-6-(4-fluorophenyl)pyridin-2- yl)oxy)-3-azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone (enantiomer 1 and 2) [0347] To a solution of Example Int.2-1-P1 (8 mg, 0.012 mmol) in DCM (4 mL) was added TFA (1 mL).
  • Example 2-1-P1 faster eluting enantiomer: MS m/z (M+H) + : calculated 580.2, observed 580.2.
  • Example Int.2-1-P2 To a solution of Example Int.2-1-P2 (10 mg, 0.015 mmol) in DCM (4 mL) was added TFA (1 mL) at rt. After 1 h the mixture was concentrated. The crude product was purified by preparative reverse-phase HPLC (20-40% MeCN/water with 0.1% TFA modifier) to give the title compound as the TFA salt.
  • Example 2-1-P2 (slower eluting enantiomer): MS m/z (M+H) + : calculated 580.6, observed 580.2.
  • the crude product was subjected to silica gel chromatography (0-60% EtOAc/petroleum ether) to give the racemate of the title compound.
  • the racemate was subjected to chiral SFC (ChiralPak AS-3, 5- 40% EtOH (with 0.05% DEA modifier)/CO2) to give the title compounds.
  • Example Int.3-1-P1 faster eluting enantiomer
  • Example Int.3-1-P2 slower eluting enantiomer
  • Step 2 ent-((1R,5S,6s)-6-((4-(2-amino-1-fluoropropan-2-yl)-6-(4-fluorophenyl)pyridin-2- yl)oxy)-3-azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone (enantiomer 1 and 2) [0350] To a solution of Example Int.3-1-P1 (58 mg, 0.087 mmol) in DCM (8 mL) was added TFA (2 mL).
  • Example 3-1-P1 faster eluting enantiomer: MS m/z (M+H) + : calculated 566.2, observed 566.4.
  • Example Int.3-1-P2 (63 mg, 0.095 mmol) in DCM (8 mL) was added TFA (2 mL) at rt. After 1 h the mixture was concentrated. The crude product was purified by preparative reverse-phase HPLC (20-40% MeCN/water with 0.1% TFA modifier) to give the title compound as the TFA salt.
  • Example 3-1-P2 (slower eluting enantiomer): MS m/z (M+H) + : calculated 566.2, observed 566.4.
  • Example 4-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)amino)-3- azabicyclo[3.1.0]hexan-3-yl)(2-cyclopropyl-8-methylimidazo[1,2-a]pyridin-6-yl)methanone
  • Step 1 tert-butyl (2- [1,2-a]pyridine-6- carbonyl)-3-azabicyclo[3.1.0]hexan-6- -6- pyridin-4-yl)propan-2- yl)carbamate
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)amino)-3- azabicyclo[3.1.0]hexan-3-yl)(2-cyclopropyl-8-methylimidazo[1,2-a]pyridin-6-yl)methanone [0353] To a solution of tert-butyl (2-(2-(((1R,5S,6s)-3-(2-cyclopropyl-8-methylimidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)amino)-6-(4-fluorophenyl)pyridin-4- yl)propan-2-yl)carbamate (39 mg, 0.062 mmol) in DCM (0.5 mL) was added HCl (4M in 1,4- dioxane,
  • Example 5-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(8-(difluoromethoxy)-2-(difluoromethyl)imidazo[1,2-a]pyridin- 6-yl)methanone
  • Step 1 benzyl (2-(2-( imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4- yl)propan-2-yl)carbamate
  • 6-bromo-8-(difluoromethoxy)-2-(difluoromethyl)imidazo[1,2- a]pyridine Int.
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(8-(difluoromethoxy)-2-(difluoromethyl)imidazo[1,2-a]pyridin- 6-yl)methanone [0355] A solution of benzyl (2-(2-(((1R,5S,6s)-3-(8-(difluoromethoxy)-2- (difluoromethyl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy
  • Example 5-1 Utilizing the procedures described in Example 5-1, the following compounds were prepared substituting the appropriate reagents for 6-bromo-8-(difluoromethoxy)-2- (difluoromethyl)imidazo[1,2-a]pyridine (Int. BJ-1). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(8-cyclopropoxy-2-methylimidazo[1,2-a]pyridin-6- yl)methanone [0358] To a solution of benzyl (2-(2-(((1R,5S,6s)-3-(8-cyclopropoxy-2-methylimidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-
  • Example 6-1 Utilizing the procedures described in Example 6-1, the following compounds were prepared substituting the appropriate reagents for 6-bromo-8-cyclopropoxy-2- methylimidazo[1,2-a]pyridine (Int. BO-1). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Example 7-1 MS m/z (M+H) + : calculated 554.2, observed 554.2.
  • Example 8-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-cyclopropyl-8-methylimidazo[1,2-a]pyridin-6-yl)methanone [0362] 2-(2-(( pyridin-4- yl)propan-2-amine (Int. O-1, 60.5 mg, 0.14 mmol), 2-cyclopropyl-8-methylimidazo[1,2- a]pyridine-6-carboxylic acid (Int.
  • Example 8-1 Utilizing the procedures described in Example 8-1, the following compounds were prepared substituting the appropriate reagents for 2-cyclopropyl-8-methylimidazo[1,2- a]pyridine-6-carboxylic acid (Int. V-1). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Example 9-1 MS m/z (M+H) + : calculated 486.6, observed 486.3.
  • Example 9-1 Utilizing the procedures described in Example 9-1, the following compounds were prepared substituting the appropriate reagents for 2-methylbenzo[d]oxazole-6-carboxylic acid. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-cyclopropyl-8-ethoxyimidazo[1,2-a]pyridin-6-yl)methanone [0367] To a solution of benzyl (2-(2-(((1R,5S,6s)-3-(2-cyclopropyl-8-ethoxyimidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluoropheny
  • Example 10-1 Utilizing the procedures described in Example 10-1, the following compounds were prepared substituting the appropriate reagents for 2-cyclopropyl-8-ethoxyimidazo[1,2-a]pyridine- 6-carboxylic acid (Int. W-4). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(1,2,4-oxadiazol-3-yl)imidazo[1,2-a]pyridin-6- yl)methanone [0370] A solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8-(1,2,4- oxadiazol-3-yl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.
  • Example 11-1 MS m/z (M+H) + : calculated 554.2, observed 554.2.
  • Example 11-1 Utilizing the procedures described in Example 11-1, the following compounds were prepared substituting the appropriate reagents for 2-methyl-8-(1,2,4-oxadiazol-3-yl)imidazo[1,2- a]pyridine-6-carboxylic acid (Int. AQ-1). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone [0373] To a solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8- (trifluoromethyl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin- 4-yl)propan-2-yl)carbamate (800 mg, 1.16 mmol) in THF (10 mL) was added TEA (0.649 mL, 4.65 mmol), Et3SiH
  • Example 12-1 MS m/z (M+H) + : calculated 554.2, observed 554.3.
  • Example 12-1 Utilizing the procedures described in Example 12-1, the following compounds were prepared substituting the appropriate reagents for 2-methyl-8-(trifluoromethyl)imidazo[1,2- a]pyridine-6-carboxylic acid (Int. V-4). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 ent-((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(7-(1-hydroxyethyl)-2-methylpyrazolo[1,5-a]pyridin-5- yl)methanone (enantiomer 1 and 2) [0376]
  • Example Int.13-1-P1 45 mg, 0.068 mmol
  • concentrated HCl (1 mL) was heated to 80 °C. After 10 minutes the mixture was cooled to rt then made basic by careful addition of solid NaHCO3.
  • Example 13- 1-P1 faster eluting enantiomer: MS m/z (M+H) + : calculated 530.6, observed 530.2.
  • Example Int.13-1-P2 (45 mg, 0.068 mmol) in concentrated HCl (1 mL) was heated to 80 °C. After 10 minutes the mixture was cooled to rt then made basic by careful addition of solid NaHCO 3 . The resulting mixture was diluted with MeOH (2 mL) then filtered then the filtrate was concentrated. The crude product was purified by preparative reverse-phase HPLC (25-55% MeCN/water with 0.1% TFA modifier) to give the title compound as the TFA salt.
  • Example 13- 1-P2 (slower eluting enantiomer): MS m/z (M+H) + : calculated 530.6, observed 530.2.
  • Example 13-1 Utilizing the procedures described in Example 13-1, the following compounds were prepared substituting the appropriate reagents for 7-(1-hydroxyethyl)-2-methylpyrazolo[1,5- a]pyridine-5-carboxylic acid (Int. AZ-1). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Example 16-1-P2 (slower eluting enantiomer): MS m/z (M+H) + : calculated 540.2, observed 540.2.
  • 1 H-NMR 500 MHz, CD3OD
  • Example 16-1 Utilizing the procedures described in Example 16-1, the following compounds were prepared substituting the appropriate reagents for 2-cyclopropyl-8-methylimidazo[1,2- a]pyridine-6-carboxylic acid (Int. V-1). Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases. Ex # R Observed Comments + [M+H] + n n . 3, 2) n n . 3, 2) n n .
  • Step 2 benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-formylimidazo[1,2-a]pyridine-6- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate [0385] To a solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-vinylimidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate (80 mg, 0.13 mmol) and NMO (29.7 mg, 0.25)
  • Step 3 benzyl (2-(2-(((1R,5S,6s)-3-(2-(difluoromethyl)imidazo[1,2-a]pyridine-6-carbonyl)-3- azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate [0386] To a solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-formylimidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate (30 mg, 0.047 mmol) in DCM (2 mL) was added DAST (0.031 mL, 0.24 mmol) at -78 °C.
  • Step 4 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-(difluoromethyl)imidazo[1,2-a]pyridin-6-yl)methanone [0387] A solution of benzyl (2-(2-(((1R,5S,6s)-3-(2-(difluoromethyl)imidazo[1,2-a]pyridine-6- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2- yl)carbamate (15 mg, 0.023 mmol) in concentrated HCl (0.5 mL) was heated to 80°C.
  • Example 18-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-(hydroxymethyl)imidazo[1,2-a]pyridin-6-yl)methanone 25700
  • Step 1 benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-(hydroxymethyl)imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2- yl)carbamate
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-(hydroxymethyl)imidazo[1,2-a]pyridin-6-yl)methanone [0389]
  • Example 18-1 MS m/z (M+H) + : calculated 502.2, observed 502.5.
  • Example 19-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methoxyimidazo[1,2-a]pyridin-6-yl)methanone
  • Step 1 benzyl (2-(2- [1,2-a]pyridine-6- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate [0390] A mixture of Cs 2 CO 3 (47.6 mg, 0.15 mmol), benzyl (2-(2-(((1R,5S,6s)-3-(2- bromoimidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4- 25700 fluor
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methoxyimidazo[1,2-a]pyridin-6-yl)methanone [0391] To a solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methoxyimidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate (23 mg, 0.036 mmol) in THF (5 mL) was added TEA (14.64 mg, 0.15 mmol), Et 3 SiH (21.04 mg, 0.18 mmol) and Pd
  • Example 20-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2,4-dimethylbenzo[d]thiazol-6-yl)methanone
  • Step 1 benzyl (2-(2-(( carbonyl)-3- azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate [0392] To a mixture of benzyl (2-(2-(((1R,5S,6s)-3-(4-bromo-2-methylbenzo[d]thiazole-6- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2,4-dimethylbenzo[d]thiazol-6-yl)methanone [0393] A solution of benzyl (2-(2-(((1R,5S,6s)-3-(2,4-dimethylbenzo[d]thiazole-6-carbonyl)-3- azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate (73 mg, 0.11 mmol) in concentrated HCl (0.5 mL, 6.09 m
  • Step 2 benzyl (2-(2-(((1R,5S,6s)-3-(4-carbamoyl-2-methylbenzo[d]thiazole-6-carbonyl)-3- azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate 25700 [0395] To a solution of benzyl (2-(2-(((1R,5S,6s)-3-(4-cyano-2-methylbenzo[d]thiazole-6- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2- yl)carbamate (127 mg, 0.19 mmol) in MeOH (8 mL) was added H 2 O 2 (21 mg, 0.211 mmol, 30 wt%) and 1M NaOH (0.3
  • Step 3 6-((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexane-3-carbonyl)-2-methylbenzo[d]thiazole-4-carboxamide
  • Example 22-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(7-cyclopropyl-2-methylpyrazolo[1,5-a]pyridin-5-yl)methanone
  • Step 1 benzyl (2-(2-(([1,5-a]pyridine-5- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2- yl)carbamate [0397] A mixture of benzyl (2-(2-(((1R,5S,6s)-3-(7-bromo-2-methylpyrazolo[1,5-a]pyridine-5- carbonyl)-3-azabicyclo[3.1.0]hexan-6
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(7-cyclopropyl-2-methylpyrazolo[1,5-a]pyridin-5-yl)methanone [0398] To a solution of benzyl (2-(2-(((1R,5S,6s)-3-(7-cyclopropyl-2-methylpyrazolo[1,5- a]pyridine-5-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4- yl)propan-2-yl)carbamate (80 mg, 0.12 mmol) in THF (2 mL) was added TEA (0.02 mL, 0.12 mmol), Et3
  • Example 22-1 MS m/z (M+H) + : calculated 526.6, observed 526.2.
  • Example 23-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(7-(2-hydroxypropan-2-yl)-2-methylpyrazolo[1,5-a]pyridin-5- yl)methanone
  • Step 1 1-(5-( pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexane-3-carbonyl)-2-methylpyrazolo[1,5-a]pyridin-7-yl)ethan-1-one [0399] To a solution of benzyl (2-(2-(((1R,5S,6s)-3-(7-bromo-2-methylpyrazolo[1,5-a]pyridine- 5-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(7-(2-hydroxypropan-2-yl)-2-methylpyrazolo[1,5-a]pyridin-5- yl)methanone [0400] To a mixture of 1-(5-((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin- 2-yl)oxy)-3-azabicyclo[3.1.0]hexane-3-carbonyl)-2-methylpyrazolo[1,5-a]pyridin-7-yl)
  • Step 2 ent-benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(8-(1-hydroxyethyl)imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2- yl)carbamate (enantiomer 1 and 2) [0401] To a mixture of benzyl (2-(2-(((1R,5S,6s)-3-(8-acetylimidazo[1,2-a]pyridine-6- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2- yl)carbamate (enantiomer 1 and 2) [0401] To a mixture of benzyl (2-(2-(((1R,5S,6
  • Example 24-1-P1 faster eluting enantiomer: MS m/z (M+H) + : calculated 516.2, observed 516.2.
  • Example Int.24-1-P2 To a solution of Example Int.24-1-P2 (30 mg, 0.046 mmol) in THF (1 mL) was added TEA (0.1 mL, 0.12 mmol), Et3SiH (27 mg, 0.23 mmol) and PdCl2 (3 mg, 0.018 mmol) at rt. After 1 h the mixture was filtered then the filtrate was concentrated. The crude product was purified by preparative reverse-phase HPLC (15-45% MeCN/water with 0.1% TFA modifier) to give the title compound as the TFA salt.
  • Example 24-1-P2 (slower eluting enantiomer): MS m/z (M+H) + : calculated 516.2, observed 516.2.
  • Example 25-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(1H-1,2,4-triazol-3-yl)imidazo[1,2-a]pyridin-6- yl)methanone
  • Step 1 6-((1R,5S,6s)- 2-yl)-6-(4- fluorophenyl)pyridin-2-yl)oxy)-3-azabicyclo[3.1.0]hexane-3-carbonyl)-2-methylimidazo[1,2- a]pyridine-8-carboxylic acid [0404] To a mixture of benzyl (2-(2-(((1R,5S,6s)-3-(8-bromo-2-methylimidazo[1,2-a]pyridine- 6-carbonyl)
  • Step 3 benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8-(1H-1,2,4-triazol-3- yl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4- yl)propan-2-yl)carbamate [0406] To a suspension of benzyl (2-(2-(((1R,5S,6s)-3-(8-carbamoyl-2-methylimidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4- yl)propan-2-yl)carbamate (50 mg, 0.075 mmol) in toluene (3 mL) was added DMF-DMA (
  • Step 4 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(1H-1,2,4-triazol-3-yl)imidazo[1,2-a]pyridin-6- yl)methanone [0407] A solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8-(1H-1,2,4- triazol-3-yl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4- yl)propan-2-yl)carbamate (35 mg, 0.051 mmol) in concentrated HCl (3 mL) was heated to
  • Example 26-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(2H-1,2,3-triazol-2-yl)imidazo[1,2-a]pyridin-6- yl)methanone N N F N Step 1: benzyl (2-(2- 1,2,3-triazol-2- yl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4- yl)propan-2-yl)carbamate [0408] An oven-dried vial was equipped with a magnetic stir bar and charged with Pd2(dba)3 (6.6 mg, 7.16 ⁇ mol) and Me 4 tBuXPhos (6.9 mg, 0.0
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(2H-1,2,3-triazol-2-yl)imidazo[1,2-a]pyridin-6- yl)methanone [0409] To a solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8-(2H-1,2,3- triazol-2-yl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4- yl)propan-2-yl)carbamate (50 mg, 0.073 mmol) in THF (10 mL) was added
  • Example 26-1 MS m/z (M+H) + : calculated 553.2, observed 553.2.
  • Example 27-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(1H-1,2,3-triazol-4-yl)imidazo[1,2-a]pyridin-6- yl)methanone
  • Step 1 benzyl (2-(2- (2-(tetrahydro-2H-pyran- 2-yl)-2H-1,2,3-triazol-4-yl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6- yl)oxy)pyridin-4-yl)propan-2-yl)carbamate [0410] To a mixture of benzyl (2-(2-(((1R,5S,6s)-3-(8-bromo-2-methyl
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(1H-1,2,3-triazol-4-yl)imidazo[1,2-a]pyridin-6- yl)methanone [0411] A solution of benzyl (2-(2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8-(2- (tetrahydro-2H-pyran-2-yl)-2H-1,2,3-triazol-4-yl)imid
  • Example 28-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(8-ethyl-2-methylimidazo[1,2-a]pyridin-6-yl)methanone [0412] Benzyl (2-(2-( pyridine-6-carbonyl)-3- azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2-yl)carbamate (Int.
  • Example 29-1 Utilizing the procedures described in Example 29-1, the following compounds were prepared substituting the appropriate reagents for 2-(tri-n-butylstannyl)oxazole. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Example 30-1 Utilizing the procedures described in Example 30-1, the following compounds were prepared substituting the appropriate reagents for azetidin-2-one. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 tert-butyl (1R,5S,6s)-6-((4-cyano-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexane-3-carboxylate [0418] To a mixture of tert-butyl (1R,5S,6s)-6-((6-chloro-4-cyanopyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexane-3-carboxylate (300 mg, 0.893 mmol) and (4-fluorophenyl)boronic acid (188 mg, 1.340 mmol) in water (0.5 mL) and 1,4-dioxane (5 mL) was added Cs2CO3 (582 mg, 1.79 mmol).
  • Step 3 2-(((1R,5S,6s)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4- fluorophenyl)isonicotinonitrile [0419] To a solution of tert-butyl (1R,5S,6s)-6-((4-cyano-6-(4-fluorophenyl)pyridin-2-yl)oxy)- 3-azabicyclo[3.1.0]hexane-3-carboxylate (150 mg, 0.38 mmol) in DCM (5 mL) was added TFA (2 mL, 26.0 mmol) at 0 °C then the mixture was warmed to rt.
  • Step 4 2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8-(trifluoromethyl)imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)isonicotinonitrile [0420] To a solution of 2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridine-6-carboxylic acid (Int.
  • Step 6 2-(4-fluorophenyl)-6-(((1R,5S,6s)-3-(2-methyl-8-(trifluoromethyl)imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)isonicotinimidamide
  • Example 32-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-cyclopropyl-7-methoxypyrazolo[1,5-a]pyridin-5- yl)methanone
  • Step 1 ((1R,5S,6s)-6-( 2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(7-bromo-2-cyclopropylpyrazolo[1,5-a]pyridin-5-yl)methanone
  • To a solution of 7-bromo-2-cyclopropylpyrazolo[1,5-a]pyridine-5-carboxylic acid (Int.
  • Step 3 tert-butyl (2-(2-(((1R,5S,6s)-3-(2-cyclopropyl-7-methoxypyrazolo[1,5-a]pyridine-5- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2- yl)carbamate
  • Tert-butyl (2-(2-(((1R,5S,6s)-3-(7-bromo-2-cyclopropylpyrazolo[1,5-a]pyridine-5- carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4-yl)propan-2- yl)carbamate (28 mg, 0.041 mmol), t-BuXphos Pd G3 (6.4 mg, 8.1 ⁇ M), and Cs 2
  • Step 4 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-fluorophenyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-cyclopropyl-7-methoxypyrazolo[1,5-a]pyridin-5- yl)methanone [0426] To a solution of tert-butyl (2-(2-(((1R,5S,6s)-3-(2-cyclopropyl-7-methoxypyrazolo[1,5- a]pyridine-5-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)-6-(4-fluorophenyl)pyridin-4- yl)propan-2-yl)carbamate (5 mg, 8 ⁇ mol) in DCM (0.2 mL) was added HCl (4N in 1,4-diox
  • Example 33-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(p-tolyl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone 25700 [0427] Benzyl (2-(2- - imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate (Int.
  • Example 33-1 Utilizing the procedures described in Example 33-1, the following compounds were prepared substituting the appropriate reagents for 4-methylphenylboronic acid. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4,4-dimethylcyclohex-1-en-1-yl)pyridin-2- yl)oxy)-3-azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone [0430] Benzyl (2-(2-(4,4-dimethylcyclohex-1-en-1-yl)-6-(((1R,5S,6s)-3-(2-methyl-8- (trifluoromethyl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin- 4-
  • Example 34-1 Utilizing the procedures described in Example 34-1, the following compounds were prepared substituting the appropriate reagents for (4,4-dimethylcyclohex-1-en-1-yl)boronic acid. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Example 35-1 Utilizing the procedures described in Example 35-1, the following compounds were prepared substituting the appropriate reagents for 2-(4,4-difluorocyclohex-1-en-1-yl)-4,4,5,5- tetramethyl-1,3,2-dioxaborolane. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Step 2 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(7,7-difluorobicyclo[4.2.0]octa-1(6),2,4- trien-3-yl)pyridin-2-yl)oxy)-3-azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8- (trifluoromethyl)imidazo[1,2-a]pyridin-6-yl)methanone [0436] Triethylsilane (39.3 ⁇ l, 0.25 mmol) was added to a mixture of benzyl (2-(2-(7,7- difluorobicyclo[4.2.0]octa-1(6),2,4-trien-3-yl)-6-(((1R,5S,6s)-3-(2-methyl-8- (trifluoromethyl)imidazo[1,2-a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hex
  • Example 37-1 ((1R,5S,6s)-6-((4-(2-aminopropan-2-yl)-6-(4-methyl-1H-indol-1-yl)pyridin-2-yl)oxy)-3- azabicyclo[3.1.0]hexan-3-yl)(2-methyl-8-(trifluoromethyl)imidazo[1,2-a]pyridin-6- yl)methanone [0437] Benzyl (2-(2-chloro-6-(((1R,5S,6s)-3-(2-methyl-8-(trifluoromethyl)imidazo[1,2- a]pyridine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)oxy)pyridin-4-yl)propan-2-yl)carbamate (Int.
  • Example 37-1 Utilizing the procedures described in Example 37-1, the following compounds were prepared substituting the appropriate reagents for 4-methyl-1H-indole. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • Example 38-1 MS m/z (M+H) + : calculated 611.6, observed 611.5.
  • Example 38-1 Utilizing the procedures described in Example 38-1, the following compounds were prepared substituting the appropriate reagents for 4-(trifluoromethyl)piperidine, HCl. Following HPLC purification, Examples were concentrated directly to give TFA or HCl salts or extracted under basic conditions to give free bases.
  • RSV-A2-GFP virus [0442] GFP sequence was derived from pJTITM R4 Dest CMV N-EmGFP pA Vector (Invitrogen) and was cloned into the intergenic sequence between wild-type RSV-A2 P and M genes using standard recombineering techniques.
  • Recombinant RSV-A2-GFP was propagated in Hep2 cells (human cells contain HeLa marker chromosomes and were derived via HeLa contamination, ATCC CCL-23) with a Multiplicity of Infection (MOI) of 0.1.
  • Virus was harvested 3 days after infection by collecting all culture material and then freeze-thawing the mixture for 5 minutes.
  • Working virus stocks were generated by thawing frozen viral cultures in 37 °C water bath, which were centrifuged at 218 g for 15 min at 4°C.1/10 of 10x SPG (Biological Industries 06-3061-01-5A) were added and then mixed. The supernatant was aliquoted, frozen in liquid nitrogen, and then transferred to a -80 °C freezer for storage.
  • Virus 25700 titer was determined by automated plaque assay in HEp-2 cells following methods described previously (Wen Z. et al.; 2019). Generating Calu-1 assay ready freeze-down (ARF): [0443] Calu-1 cells were purchased from ATCC (Cat# HTB-54) and were expanded in growth media (DMEM containing 10%FBS, 100U/ml Penicillin-Streptomycin). To make ARF, cell culture media was removed and discarded and the cell layer was briefly rinsed with PBS to remove serum.2.5 mL of TrypLE Express solution were added and cells incubated until dislodged, to which growth media was added and the cells were resuspended by gentle pipetting.
  • ARF Calu-1 assay ready freeze-down
  • Cells were counted for concentration and viability as determined by ViCell. Cells were centrifuged at RT, 300 g for 5 minutes to pellet cells. The supernatant was gently aspirated and the pellet was flicked to loosen cells. Cells were resuspended in an appropriate volume of freezing medium (DMEM containing 10% DMSO, 10%FBS, 100U/ml Penicillin-Streptomycin) to achieve a concentration of 5 x 10 6 cells/mL.1 mL aliquots of the cell suspension were transferred to freezing vials.
  • DMEM containing 10% DMSO, 10%FBS, 100U/ml Penicillin-Streptomycin
  • RSV-A2-GFP viral replication assay [0444] Assay ready freeze-down (ARF, generation described separately) Calu-1 cells (ATCC HTB-54) were used. Compound plates were prepared by dispensing compounds dissolved in DMSO into the wells of a 384 well Corning® 3985 plate with an ECHO acoustic dispenser and compounds were tested in 10-point serial 3-fold dilution. Wells with DMSO (final concentration of 0.4 %) or a compound at a concentration at which viral replication was completely inhibited with a control compound were used as viral replication assay Min_E and Max_E control, respectively.
  • CTG CellTiter-Glo
  • the cells were dispensed at 50 ⁇ L/well using Bravo with 50 ⁇ L filtered tips into compound plates. Plates were covered with MicroClime lids, loaded with 7.5 mL of assay media to minimize evaporation, and were incubated at 37 °C and 5% CO2 for 96 hrs. Following incubation, distinct, GFP-expressing cells were counted using an Acumen imaging system with appropriate settings. A same-well CTG assay was performed by adding 10 ⁇ L/well reconstituted 25700 CellTiter-Glo reagent (Promega G7573) and plates were read on PerkinElmer Envision according to manufacturer’s instructions. Raw data were loaded and analyzed in ActivityBase.
  • Antiviral IC 50 and cytotoxicity CC 50 values were determined using a 4 parameter logistic fit based on the Levenberg-Marquardt algorithm. Model: 205 - 4 Parameter Logistic.
  • Generation and propagation of hMPV-GFP virus [0445] GFP expressing rgHMPV#3 p3 was generated by the Buchholz lab (Biacchesi S. et al., J Virol.2007 Jun; 81(11): 6057–6067) and propagated in VERO cells (ATCC Cat# CCL-81) with a Multiplicity of Infection (MOI) of 0.1. Virus was harvested 4 days after infection by collecting cultured material and was freeze-thaw cycled in liquid nitrogen twice.
  • MOI Multiplicity of Infection
  • Working virus stocks were generated from thawing frozen samples in 37 °C water bath, which were centrifuged at 218 g for 15 min at 4°C.1/10 of 10x SPG (Biological Industries 06-3061-01-5A) were added and then mixed. The supernatant was aliquoted, frozen in liquid nitrogen, and then transferred to a -80 °C freezer for storage. Virus titer was determined by performing titration test in 96-well plates and calculate approximate virus titer using GFP event/well data.
  • hMPV-GFP viral replication assay [0446] Compound plates were prepared by dispensing compounds dissolved in DMSO into wells of a 384 well Corning 3985 polystyrene flat clear bottom optical imaging microplate (202.5nL/well) using an ECHO acoustic dispenser. Each compound was tested in10-point serial 3-fold dilution (typical final concentrations: 40,300nM – 2 nM). Wells with DMSO (final concentration of 0.4 %) or a compound at a concentration at which viral replication was completely inhibited relative to a control compound were used as viral replication assay Min_E and Max_E control, respectively.
  • VERO cells Continuous culture of VERO cells was maintained in complete culture media (OptiMEM supplemented with 2mM GlutaMAXTM and 100U/ml Penicillin- Streptomycin).
  • VERO cells were trypsinized with 0.25 % Trypsin-EDTA until cells were dislodged, then cells were re-suspended with 1 mL FBS. Cells were spundown at 300g for 5 minutes and cells were washed twice with culture media and counted using default parameters on ViCell. Cells were then diluted to 100,000 cells/mL (5,000 cells/50 ⁇ L) in complete culture media + TrypLE Select (80 ⁇ L/mL).
  • a same-well CTG assay was performed by adding 10 ⁇ L/well reconstituted CellTiter-Glo reagent (Promega G7573) and plates were read on PerkinElmer Envision according to manufacturer’s instructions. Raw data were loaded and analyzed in ActivityBase. Antiviral IC50 and cytotoxicity CC50 values were determined using a 4 parameter logistic fit based on the Levenberg-Marquardt algorithm. Model: 205 - 4 Parameter Logistic. References: Wen Z, Citron M, Bett AJ, Espeseth AS, Vora KA, Zhang L, DiStefano DJ. Development and application of a higher throughput RSV plaque assay by immunofluorescent imaging. J Virol Methods.2019 Jan;263:88-95.

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Abstract

La présente divulgation concerne des composés de formule I : (I) et leur utilisation en tant qu'agents antiviraux pour l'inhibition de la réplication du VRSh et du MPVh et le traitement et la prophylaxie de l'infection par VRSh et MPVh.
PCT/US2024/034649 2023-06-26 2024-06-20 Inhibiteurs du virus respiratoire syncytial humain et du métapneumovirus WO2025006293A2 (fr)

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