WO2024039861A1 - 2,3,4,9-tetrahydro-1h-pyrido[3,4-b]indole derivatives as estrogen receptor modulators for the treatment of cancer - Google Patents

2,3,4,9-tetrahydro-1h-pyrido[3,4-b]indole derivatives as estrogen receptor modulators for the treatment of cancer Download PDF

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WO2024039861A1
WO2024039861A1 PCT/US2023/030599 US2023030599W WO2024039861A1 WO 2024039861 A1 WO2024039861 A1 WO 2024039861A1 US 2023030599 W US2023030599 W US 2023030599W WO 2024039861 A1 WO2024039861 A1 WO 2024039861A1
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compound
mmol
methyl
cancer
fluoro
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PCT/US2023/030599
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French (fr)
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Brian R. Hearn
Xufeng Cao
David YEGHIKYAN
Rupa S. Shetty
Brian RICH
Reena CHAWLA
Dirk A. Heerding
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Olema Pharmaceuticals, Inc.
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Publication of WO2024039861A1 publication Critical patent/WO2024039861A1/en

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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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems

Definitions

  • the estrogen receptor (ER) plays important roles in various diseases, disorders, and conditions, such as cancers, including breast cancers, menopause-related conditions or symptoms, and osteoporosis.
  • About 70% of human breast cancers are hormone dependent and ER-positive.
  • a variety of treatments have been developed to target the estrogen receptor and/or its activities.
  • Selective estrogen receptor modulators and/or degraders (SERDs) are a particularly useful or promising tools for such therapy.
  • an estrogen receptor modulator that acts as an agonist (or partial agonist) in bone tissue may be useful for treating osteoporosis, e.g., in post-menopausal women.
  • an estrogen receptor modulator that acts as an antagonist in breast tissue may be useful for treating breast cancer. In some instances, the same estrogen receptor modulator may be used in both scenarios.
  • the present disclosure provides compounds that are estrogen receptor modulators.
  • provided compounds are estrogen receptor agonists, e.g., as defined herein.
  • provided compounds are estrogen receptor antagonists, e.g., as defined herein.
  • estrogen receptors include those coded for by both wild-type and mutant versions (e.g., those containing activating mutations) of the gene encoding Estrogen Receptor-alpha (ERa), Estrogen Receptor 1 (ESRI).
  • the estrogen receptor is a tripartite protein comprising two distinct transcriptional activation functions (AF1 and AF2). Complete anti-estrogen activity requires inactivation of both AF1 and AF2. Activating mutations in the gene that codes for estrogen receptor 1 allows for activation of both AF1 and AF2 even in the absence of estrogen.
  • the present disclosure provides certain compounds and compositions that are complete estrogen receptor antagonists, and therefore do not suffer from the deficiencies found in previous therapies.
  • provided compounds may be orally bioavailable.
  • the present disclosure provides an estrogen receptor modulator (e.g., an estrogen receptor agonist, an estrogen receptor antagonist, and/or a complete estrogen receptor antagonist) that is a compound of Formula I: or a pharmaceutically acceptable salt thereof, wherein A, B, L, n, R 1 , R 2 , R 3 , and R 4 are as defined herein.
  • an estrogen receptor modulator e.g., an estrogen receptor agonist, an estrogen receptor antagonist, and/or a complete estrogen receptor antagonist
  • the present disclosure provides methods of treating a disease, disorder, or condition associated with an estrogen receptor. In some embodiments, the present disclosure provides a methods of treating a disease, disorder, or condition associated with a mutation of an estrogen receptor.
  • the present disclosure provides methods of treating a cancer.
  • the present disclosure provides methods of treating a cancer comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof, optionally in combination with an anti-cancer agent.
  • the present disclosure provides methods of preventing recurrence of a cancer. In some embodiments, the present disclosure provides methods of preventing recurrence of a cancer comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof, optionally in combination with an anti-cancer agent.
  • the present disclosure provides methods of treating osteoporosis, e.g., in post-menopausal women. In some embodiments, the present disclosure provides methods of treating osteoporosis comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof.
  • the present disclosure provides methods of treating one or more menopausal symptoms or conditions. In some embodiments, the present disclosure provides methods of treating one or more menopausal symptoms comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof.
  • the present disclosure provides compounds and compositions useful as estrogen receptor modulators (e.g., estrogen receptor agonists, estrogen receptor antagonists, and/or complete estrogen receptor antagonists).
  • estrogen receptor modulators e.g., estrogen receptor agonists, estrogen receptor antagonists, and/or complete estrogen receptor antagonists.
  • such compounds include those of the formulae described herein, or a pharmaceutically acceptable salt thereof, wherein each variable is as defined and described herein.
  • structures depicted herein are meant to include all stereoisomeric (e.g., enantiomeric or diastereomeric) forms of the structure, as well as all geometric or conformational isomeric forms of the structure.
  • the R and S configurations of each stereocenter are contemplated as part of the disclosure. Therefore, single stereochemical isomers, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of provided compounds are within the scope of the disclosure.
  • Table 1 shows one or more stereoisomers of a compound, and unless otherwise indicated, represents each stereoisomer alone and/or as a mixture. Unless otherwise stated, all tautomeric forms of provided compounds are within the scope of the disclosure.
  • structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures including replacement of hydrogen by deuterium or tritium, or replacement of a carbon by 13 C- or 14 C-enriched carbon are within the scope of this disclosure.
  • the term "approximately” or “about” may encompass a range of values that are within (i.e., ⁇ ) 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.
  • Administering typically refers to the administration of a composition to a subject to achieve delivery of an agent that is, or is included in, a composition to a target site or a site to be treated.
  • agents that are, or is included in, a composition to a target site or a site to be treated.
  • routes that may, in appropriate circumstances, be utilized for administration to a subject, for example a human.
  • administration may be ocular, oral, parenteral, topical, etc.
  • administration may be bronchial (e.g., by bronchial instillation), buccal, dermal (which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreal, etc.
  • bronchial e.g., by bronchial instillation
  • buccal which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal, etc.
  • enteral intra-arterial, intradermal, intragas
  • administration may be parenteral. In some embodiments, administration may be oral. In some particular embodiments, administration may be intravenous. In some particular embodiments, administration may be subcutaneous. In some embodiments, administration may involve only a single dose. In some embodiments, administration may involve application of a fixed number of doses. In some embodiments, administration may involve dosing that is intermittent (e.g., a plurality of doses separated in time) and/or periodic (e.g., individual doses separated by a common period of time) dosing. In some embodiments, administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time. In some embodiments, administration may comprise a prime-and-boost protocol. A prime-and-boost protocol can include administration of a first dose of a pharmaceutical composition followed by, after an interval of time, administration of a second or subsequent dose of a pharmaceutical composition.
  • a prime-and-boost protocol can include administration of a first dose of a pharmaceutical composition followed by, after
  • agonist generally refers to an agent whose presence or level correlates with elevated level or activity of a target, as compared with that observed absent the agent (or with the agent at a different level).
  • an agonist is one whose presence or level correlates with a target level or activity that is comparable to or greater than a particular reference level or activity (e.g., that observed under appropriate reference conditions, such as presence of a known agonist, e.g., a positive control).
  • an agonist may be a direct agonist in that it exerts its influence directly on (e.g., interacts directly with) the target; in some embodiments, an agonist may be an indirect agonist in that it exerts its influence indirectly (e.g., by acting on, such as interacting with, a regulator of the target, or with some other component or entity.
  • Aliphatic refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “cycloaliphatic”), that has a single point or more than one points of attachment to the rest of the molecule.
  • aliphatic groups contain 1-12 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms (e.g., Ci-e).
  • aliphatic groups contain 1-5 aliphatic carbon atoms (e.g., C1-5). In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms (e.g., C1-4). In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms (e.g., C1-3), and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms (e.g., C1-2). Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups and hybrids thereof. A preferred aliphatic group is C1-6 alkyl.
  • Alkyl refers to a saturated, optionally substituted straight or branched chain hydrocarbon group having (unless otherwise specified) 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms (e.g., C1-12, C1-10, Ci-s, C1-6, Ci-4, Ci- 3, or C1-2).
  • exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, and heptyl.
  • Alkylene refers to a bivalent alkyl group. In some embodiments, “alkylene” is a bivalent straight or branched alkyl group. In some embodiments, an "alkylene chain" is a polymethylene group, i.e., -(CH2)n-, wherein n is a positive integer, e.g., from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3.
  • An optionally substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms is optionally replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group and also include those described in the specification herein.
  • two substituents of the alkylene group may be taken together to form a ring system.
  • two substituents can be taken together to form a 3- to 7- membered ring.
  • the substituents can be on the same or different atoms.
  • the suffix “-ene” when appended to certain groups herein are intended to refer to a bifunctional moiety of said group.
  • “-ene”, when appended to “cyclopropyl” becomes “cyclopropylene” and is intended to refer to a bifunctional cyclopropyl group, e.g.,
  • Alkenyl refers to an optionally substituted straight or branched chain or cyclic hydrocarbon group having at least one double bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms(e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or C2-3).
  • alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl.
  • cycloalkenyl refers to an optionally substituted non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon double bond and having about 3 to about 10 carbon atoms.
  • exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.
  • Alkynyl refers to an optionally substituted straight or branched chain hydrocarbon group having at least one triple bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms (e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or C2-3).
  • exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.
  • Antagonist generally refers to an agent whose presence or level correlates with decreased level or activity of a target, as compared with that observed absent the agent (or with the agent at a different level).
  • an antagonist is one whose presence or level correlates with a target level or activity that is comparable to or less than a particular reference level or activity (e.g., that observed under appropriate reference conditions, such as presence of a known antagonist, e.g., a positive control).
  • an antagonist may be a direct antagonist in that it exerts its influence directly on (e.g., interacts directly with) the target; in some embodiments, an antagonist may be an indirect antagonist in that it exerts its influence indirectly (e.g., by acting on, such as interacting with, a regulator of the target, or with some other component or entity.
  • Aryl refers to monocyclic and bicyclic ring systems having a total of six to fourteen ring members (e.g., C6-C14), wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. In some embodiments, an “aryl” group contains between six and twelve total ring members (e.g., C6-C12). The term “aryl” may be used interchangeably with the term “aryl ring”. In certain embodiments, “aryl” refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents.
  • aryl groups are hydrocarbons.
  • an “aryl” ring system is an aromatic ring (e.g., phenyl) that is fused to a non-aromatic ring (e.g., cycloalkyl). Examples of aryl rings include that are fused include [0029]
  • Biological sample typically refers to a sample obtained or derived from a biological source (e.g., a tissue or organism or cell culture) of interest, as described herein.
  • a source of interest comprises an organism, such as an animal or human.
  • a biological sample is or comprises biological tissue or fluid.
  • a biological sample may be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; cell-containing body fluids; free floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; washings or lavages such as a ductal lavages or broncheoalveolar lavages; aspirates; scrapings; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions, and/or excretions; and/or cells therefrom, etc.
  • a biological sample is or comprises cells obtained from an individual.
  • obtained cells are or include cells from an individual from whom the sample is obtained.
  • a sample is a “primary sample” obtained directly from a source of interest by any appropriate means.
  • a primary biological sample is obtained by methods selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluid (e.g., blood, lymph, feces etc.), etc.
  • sample refers to a preparation that is obtained by processing (e.g., by removing one or more components of and/or by adding one or more agents to) a primary sample. For example, filtering using a semi-permeable membrane.
  • processing e.g., by removing one or more components of and/or by adding one or more agents to
  • a primary sample For example, filtering using a semi-permeable membrane.
  • Such a “processed sample” may comprise, for example, nucleic acids or proteins extracted from a sample or obtained by subjecting a primary sample to techniques such as amplification or reverse transcription of mRNA, isolation and/or purification of certain components, etc.
  • Carrier refers to a diluent, adjuvant, excipient, or vehicle with which a composition is administered.
  • carriers can include sterile liquids, such as, for example, water and oils, including oils of petroleum, animal, vegetable or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil and the like.
  • carriers are or include one or more solid components.
  • Combination therapy refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents or modality(ies)).
  • the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens.
  • “administration” of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination.
  • combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some embodiments, two or more agents, or active moieties thereof, may be administered together in a combination composition, or even in a combination compound (e.g., as part of a single chemical complex or covalent entity).
  • Comparable refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison therebetween so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed.
  • comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features.
  • composition may be used to refer to a discrete physical entity that comprises one or more specified components.
  • a composition may be of any form - e.g., gas, gel, liquid, solid, etc.
  • Cycloaliphatic refers to a monocyclic C3-8 hydrocarbon or a bicyclic C5-10 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point or more than one points of attachment to the rest of the molecule.
  • Cycloalkyl refers to an optionally substituted saturated monocyclic or polycyclic ring system of about 3 to about 10 ring carbon atoms.
  • Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.
  • Dosage form or unit dosage form may be used to refer to a physically discrete unit of an active agent (e.g., a therapeutic or diagnostic agent) for administration to a subject.
  • an active agent e.g., a therapeutic or diagnostic agent
  • each such unit contains a predetermined quantity of active agent.
  • such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen).
  • Dosing regimen or therapeutic regimen may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time.
  • a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.
  • a dosing regimen comprises a plurality of doses each of which is separated in time from other doses.
  • individual doses are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses.
  • all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).
  • Excipient refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stabilizing effect.
  • suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.
  • heteroaliphatic or “heteroaliphatic group”, as used herein, denotes an optionally substituted hydrocarbon moiety having, in addition to carbon atoms, from one to five heteroatoms, that may be straight-chain (i.e., unbranched), branched, or cyclic (“heterocyclic”) and may be completely saturated or may contain one or more units of unsaturation, but which is not aromatic.
  • heteroatom refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quatemized form of a basic nitrogen.
  • nitrogen also includes a substituted nitrogen.
  • heteroaliphatic groups contain 1-10 carbon atoms wherein 1-3 carbon atoms are optionally and independently replaced with heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, heteroaliphatic groups contain 1-4 carbon atoms, wherein 1-2 carbon atoms are optionally and independently replaced with heteroatoms selected from oxygen, nitrogen, and sulfur. In yet other embodiments, heteroaliphatic groups contain 1-3 carbon atoms, wherein 1 carbon atom is optionally and independently replaced with a heteroatom selected from oxygen, nitrogen, and sulfur. Suitable heteroaliphatic groups include, but are not limited to, linear or branched, heteroalkyl, heteroalkenyl, and heteroalkynyl groups. For example, a 1- to 10 atom heteroaliphatic group includes the following exemplary groups: -O-CH3, -CH2-O-CH3, -O-CH2- CH2-O-CH2-CH2-O-CH3, and the like.
  • Heteroaryl refers to monocyclic or bicyclic ring groups having 5 to 10 ring atoms (e.g., 5- to 6-membered monocyclic heteroaryl or 9- to 10-membered bicyclic heteroaryl); having 6, 10, or 14 71-electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms.
  • Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[l,2-a]pyrimidinyl, imidazo[l,2-a]pyridyl, imidazo[4,5-b]pyridyl, imidazo[4,5-c]pyridyl, pyrrol opyridyl, pyrrolopyrazinyl, thienopyrimidinyl, triazolopyridyl, and benzois
  • heteroaryl and “heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms).
  • Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzotri azolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4/7- quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3-b]-l,4-oxazin-3(4H)-one, 4H- thieno[3,2-b]pyrrole, and benzoisoxazolyl.
  • Heteroatom refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quatemized form of a basic nitrogen.
  • Heterocycle As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a stable 3- to 8-membered monocyclic, a 6- to 10-membered bicyclic, or a 10- to 16-membered polycyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, such as one to four, heteroatoms, as defined above.
  • nitrogen includes a substituted nitrogen.
  • the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (as in N-substituted pyrrolidinyl).
  • a heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted.
  • saturated or partially unsaturated heterocyclic radicals include, without limitation, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiamorpholinyl.
  • a heterocyclyl group may be mono-, bi-, tri-, or polycyclic, preferably mono-, bi-, or tricyclic, more preferably mono- or bicyclic.
  • a bicyclic heterocyclic ring also includes groups in which the heterocyclic ring is fused to one or more aryl rings.
  • Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl.
  • a bicyclic heterocyclic ring can also be a spirocyclic ring system (e.g., 6- to 11 -membered spirocyclic fused heterocyclic ring having, in addition to carbon atoms, one or more heteroatoms as defined above (e.g., one, two, three or four heteroatoms)).
  • a bicyclic heterocyclic ring can also be a bridged ring system (e.g., 7- to 11 -membered bridged heterocyclic ring having one, two, or three bridging atoms.
  • modulator refers to a compound (e.g., a small molecule) that can alter the activity of another molecule (e.g., a protein).
  • a modulator can cause an increase or decrease in the magnitude of a certain activity of a type of molecule as compared to the magnitude of the activity in the absence of the modulator.
  • a modulator can be an agonist or an antagonist of a particular target, as those terms are defined herein.
  • a modulator is an agonist.
  • a modulator is an antagonist.
  • oral administration and “administered orally” as used herein have their art-understood meaning referring to administration by mouth of a compound or composition.
  • parenteral administration and “administered parenterally” as used herein have their art-understood meaning referring to modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal, and intrastemal injection and infusion.
  • Partially unsaturated refers to a ring moiety that includes at least one double or triple bond between ring atoms.
  • the term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (e.g., aryl or heteroaryl) moieties, as herein defined.
  • Patient or subject refers to any organism to which a provided composition is or may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients or subjects include animals (e g., mammals such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, a patient is a human. In some embodiments, a patient or a subject is suffering from or susceptible to one or more disorders or conditions. In some embodiments, a patient or subject displays one or more symptoms of a disorder or condition. In some embodiments, a patient or subject has been diagnosed with one or more disorders or conditions. In some embodiments, a patient or a subject is receiving or has received certain therapy to diagnose and/or to treat a disease, disorder, or condition.
  • animals e.g., mammals such as mice, rats, rabbits, non-human primates, and/or humans.
  • a patient is a human.
  • a patient or a subject is suffering from or susceptible to one or more disorders or conditions
  • composition refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers.
  • the active agent is present in unit dose amount appropriate for administration in a therapeutic or dosing regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population.
  • compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonary, and to other mucosal surfaces.
  • oral administration for example, drenches (aqueous or non-aqueous solutions or suspension
  • compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • compositions that are appropriate for use in pharmaceutical contexts, i.e., salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
  • Prevent or prevention when used in connection with the occurrence of a disease, disorder, and/or condition, refer to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time.
  • Substituted or optionally substituted As described herein, compounds of the invention may contain “optionally substituted” moieties.
  • the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. “Substituted” applies to one or more hydrogens that are either explicit or implicit from the structure (e.g., O RI refers to at least ).
  • an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
  • Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds.
  • stable refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes provided herein.
  • Groups described as being “substituted” preferably have between 1 and 4 substituents, more preferably 1 or 2 substituents.
  • Groups described as being “optionally substituted” may be unsubstituted or be “substituted” as described above.
  • Suitable monovalent substituents on R° are independently halogen, -(CH 2 )o- 2 R*, -(haloR*), -(CH 2 )o- 2 OH, -(CH 2 )o- 2 OR*, -(CH 2 )o-
  • Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: -O(CR* 2 ) 2-3 O-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Suitable substituents on the aliphatic group of R* include halogen, - R*, -(haloR*), -OH, -OR*, -O(haloR’), -CN, -C(O)OH, -C(O)OR’, -NH 2 , -NHR*, -NR* 2 , or -NO 2 , wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1-4 aliphatic, -CH 2 Ph, -0(CH 2 )o iPh, or a 3- to 6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include -R f , -NR f 2 , -C(O)R f , -C(O)OR f , -C(O)C(O)R f , C(O)CH 2 C(O)R t , -S(O) 2 R T , -SCO ⁇ NR ⁇ , -C(S)NR f 2 , -CCNT ⁇ NR ⁇ , or -NCR ⁇ SCO) ⁇ ; wherein each R' is independently hydrogen, C1-6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R', taken together with their intervening atom(s) form an unsubstitute
  • Suitable substituents on the aliphatic group of R? are independently halogen, - R*, -(haloR*), -OH, -OR’, -O(haloR’), -CN, -C(O)OH, -C(O)OR*, -NH 2 , -NHR*, -NR* 2 , or -NO 2 , wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C 1-4 aliphatic, -CH 2 Ph, -0(CH 2 )o-iPh, or a 3- to 6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • Small molecule means a low molecular weight organic and/or inorganic compound.
  • a “small molecule” is a molecule that is less than about 5 kilodaltons (kD) in size.
  • a small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD.
  • the small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D.
  • a small molecule is less than about 2000 g/mol, less than about 1500 g/mol, less than about 1000 g/mol, less than about 800 g/mol, or less than about 500 g/mol. In some embodiments, a small molecule is not a polymer.
  • a small molecule does not include a polymeric moiety.
  • a small molecule is not and/or does not comprise a protein or polypeptide (e.g., is not an oligopeptide or peptide).
  • a small molecule is not and/or does not comprise a polynucleotide (e.g., is not an oligonucleotide).
  • a small molecule is not and/or does not comprise a polysaccharide; for example, in some embodiments, a small molecule is not a glycoprotein, proteoglycan, glycolipid, etc.). In some embodiments, a small molecule is not a lipid.
  • a small molecule is a modulating agent (e.g., is an inhibiting agent or an activating agent).
  • a small molecule is biologically active.
  • a small molecule is detectable (e.g., comprises at least one detectable moiety).
  • a small molecule is a therapeutic agent.
  • small molecule compounds have structures that can exist in one or more steroi someric forms.
  • such a small molecule may be utilized in accordance with the present disclosure in the form of an individual enantiomer, diastereomer or geometric isomer, or may be in the form of a mixture of stereoisomers; in some embodiments, such a small molecule may be utilized in accordance with the present disclosure in a racemic mixture form.
  • small molecule compounds have structures that can exist in one or more tautomeric forms.
  • such a small molecule may be utilized in accordance with the present disclosure in the form of an individual tautomer, or in a form that interconverts between tautomeric forms.
  • small molecule compounds have structures that permit isotopic substitution (e.g., 2 H or 3 H for H; n C, 13 C or 14 C for 12 C; 13 N or 15 N for 14 N; 17 O or 18 O for 16 O; 36 C1 for 35 C1 or j7 Cl; 18 F for 19 F; 131 I for 127 I; etc.).
  • such a small molecule may be utilized in accordance with the present disclosure in one or more isotopically modified forms, or mixtures thereof.
  • reference to a particular small molecule compound may relate to a specific form of that compound.
  • a particular small molecule compound may be provided and/or utilized in a salt form (e.g., in an acid-addition or baseaddition salt form, depending on the compound); in some such embodiments, the salt form may be a pharmaceutically acceptable salt form.
  • a small molecule compound is one that exists or is found in nature
  • that compound may be provided and/or utilized in accordance in the present disclosure in a form different from that in which it exists or is found in nature.
  • a reference preparation of interest e.g., in a primary sample from a source of interest such as a biological or environmental source
  • a preparation of a single stereoisomer of a small molecule compound may be considered to be a different form of the compound than a racemic mixture of the compound; a particular salt of a small molecule compound may be considered to be a different form from another salt form of the compound; a preparation that contains only a form of the compound that contains one conformational isomer ((Z) or (E)) of a double bond may be considered to be a different form of the compound from one that contains the other conformational isomer ((E) or (Z)) of the double bond; a preparation in which one or more atoms is a different isotope than is present in a reference preparation may be considered to be a different form; etc.
  • the symbol as used herein refers to a point of attachment between two atoms. Additionally or alternatively, the symbol refers to a point of attachment ring in a spirocyclic manner
  • Treat As used herein, the terms “treat,” “treatment,” or “treating” refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition, for example, for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition.
  • the present disclosure provides compounds that are estrogen receptor modulators.
  • provided compounds are estrogen receptor agonists.
  • an “estrogen receptor agonist” refers to a compound or composition that produces an agonistic effect when contacting the estrogen receptor of a subject or biological sample.
  • an estrogen receptor agonist is characterized by having (i) at least 80% increase in the E2 -normalized signal in the AP assay (agonist mode) of Example 209 and (ii) no more than 80% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209.
  • provided compounds are estrogen receptor antagonists.
  • an “estrogen receptor antagonist” refers to a compound or composition that produces an antagonistic effect when contacting the estrogen receptor of a subject or biological sample.
  • an estrogen receptor antagonist is characterized by having:
  • the present disclosure provides compounds that are complete estrogen receptor (ER) antagonists.
  • ER complete estrogen receptor
  • a “complete estrogen receptor antagonist” refers to a compound or composition that produces an antagonistic effect when contacting the estrogen receptor of a subject or biological sample, with minimal agonistic effect (e g., with no or substantially no agonistic effect).
  • Complete estrogen receptor antagonism is determined according to methods described herein, for example in Example 209.
  • a complete estrogen receptor antagonist is characterized by having (i) a pICso greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode) of Example 209.
  • a complete estrogen receptor antagonist is characterized by having (i) a pICso greater than 7.5 and at least a 10% reduction in the E2- normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode) of Example 209.
  • a complete estrogen receptor antagonist is an agent (e.g., a small molecule compound) that shows ER antagonism and no or substantially no ER agonism in one or more of ERct protein level assays, MCF-7 cell line assays, Ishikawa cell line assays (measuring wild type ER and certain mutants including mutants lacking AF1 and/or AF2 domains), and rodent uterine weight gain assays. See, generally, WO 2017/059139.
  • a complete estrogen receptor antagonist has three characteristics: it (1) inhibits both activating function 1 (AF1) and activating function 2 (AF2), as complete anti-estrogen activity requires inactivation of both AF1 and AF2; (2) promotes ER degradation; and (3) lacks the partial ER agonist activity observed with certain other agents.
  • AF1 and AF2 activating function 1
  • AF2 activating function 2
  • the present disclosure provides a compound of Formula I: or a pharmaceutically acceptable salt thereof, wherein: A is an optionally substituted 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S or an optionally substituted 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S; L is a covalent bond or an optionally substituted bivalent group selected from -C 1 -C 6 aliphatic-, - L a -C 0 -C 5 aliphatic-, and -C 1 -C 5 aliphatic-L a -, wherein L a is selected from -S-, -SO-, -SO 2 -, and -N(R a )-; B is selected from –OH, -CO2H, C1-C6 aliphatic, 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O,
  • A is an optionally substituted 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S or an optionally substituted 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S.
  • A is 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S or 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S, each optionally substituted with halogen, -(CH2)o ⁇ R°, or -(CH 2 )O-40R°.
  • A is optionally substituted 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S optionally substituted with halogen, -(CH2)o-4R°, or -(CH 2 )o-40R°. In some embodiments, A is 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S, optionally substituted with halogen or Ci-Ce aliphatic.
  • A is optionally substituted 7-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 8-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 9- membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S.
  • A is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-[0075]
  • A is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • A is N-(0,1] n-(0,1] n-(0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1] n-[0,1]
  • A is optionally substituted 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 4- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, A is 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S optionally substituted with halogen, -(CH2)o-4R°, or -(CI l2 )o-40R°.
  • A is 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S comprising 1-4 heteroatoms selected from N, O, and S, optionally substituted with halogen or Ci-Ce aliphatic.
  • A is optionally substituted piperidinyl.
  • A is: , wherein * represents a point of attachment to moiety L.
  • L is a covalent bond or an optionally substituted bivalent group selected from Ci-Ce aliphatic, -L a -Co-C5 aliphatic-, and -C1-C5 aliphatic-!?-, wherein L a is selected from -S-, -SO-, -SO2-, and -N(R a )-.
  • L a is selected from -S-, -SO-, -SO2-, and -N(R a )-.
  • L is a covalent bond
  • L is optionally substituted Ci-Ce aliphatic.
  • L is -(CH2)I-6-.
  • L is -CH2-, -CH2-CH2-, -CH2-CH2-, -CH2- CH2-CH2-, -CH2-CH2-CH2-CH2-, -CH2-CH2-CH2-CH2-, or -CH2-CH2-CH2-CH2-CH2-CH2-.
  • L is -CH2-.
  • L is optionally substituted -L a -Co-Cs aliphatic.
  • L is -S-C0-C5 aliphatic, -SO-C0-C5 aliphatic, -SO2-C0-C5 aliphatic, or -N(R a )-Co- C5 aliphatic.
  • L is -S-C0-C5 aliphatic.
  • L is -SO-Co- Cs aliphatic.
  • L is -SO2-C0-C5 aliphatic.
  • L is - N(R a )-Co-Cs aliphatic.
  • L is L a . In some embodiments, L is -S-, -SO-, - SO2-, or -N(R a )-. In some embodiments, L is -S-. In some embodiments, L is -SO-. In some embodiments, L is -SO2-. In some embodiments, L is -N(R a )-. In some embodiments, L is - N(H)-. In some embodiments, L is -N(CH3)-.
  • L is selected from a covalent bond, -CH2-, -CH2-CH2-, -CH2- CH2-CH2-, -S-, and -SO2-. In some embodiments, L is selected from a covalent bond, -CH2-, - CH2-CH2-, -CH2-CH2-CH2-, -S-, and -N(H)-.
  • B is selected from -OH, -CO2H, Ci-Ce aliphatic, 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S, and C3-C6 cycloaliphatic.
  • n is 0.
  • B is selected from 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S and C3-C6 cycloaliphatic.
  • B is selected from -OH, -CO2H, Ci-Ce aliphatic, and 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S.
  • B is selected from -CO2H and 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S.
  • B is -OH.
  • B is -CO2H.
  • B is Ci-Ce aliphatic. In some embodiments, B is Ci-Ce alkyl.
  • B is 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is a 4- to 7-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is a 4- or 7-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S.
  • B is 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is 3-membered heterocyclyl comprising 1 heteroatom selected from N, O, and S. In some embodiments, B is 4-membered heterocyclyl comprising 1-2 heteroatoms selected from N, O, and S. In some embodiments, B is 5-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S.
  • B is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or Attorney Docket No.: 2012034-0263 morpholinyl.
  • B is azetidinyl or pyrrolidinyl.
  • B is azetidinyl.
  • B is pyrrolidinyl.
  • B is selected from: , [0093] In some embodiments, B is selected from: and . [0094] In some embodiments, B is selected from In some embodiments, B is .
  • B is 6- to 12-membered bicyclic fused or spirocyclic heterocyclyl.
  • B is 6- to 8-membered bicyclic fused or spirocyclic heterocyclyl.
  • B is 6- to 12-membered bicyclic fused heterocyclyl.
  • B is 6- to 8-membered bicyclic fused heterocyclyl.
  • B is 6- to 12-membered bicyclic spirocyclic heterocyclyl.
  • B is 6- to 8- membered bicyclic spirocyclic heterocyclyl.
  • B is selected from: , and Page 28 of 231 11528310v1 [0096] In some embodiments, B is selected from:
  • B is selected from:
  • B is:
  • B is Ca-Ce cycloaliphatic In some embodiments, B is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
  • each R 4 is independently oxo, halogen, -CN, -OR a , - N(R a ) 2 , -C(O)R a , -OC(O)R a , -C(O)OR a , -C(O)N(R a ) 2 , -N(R a )C(O)R a , or an optionally substituted group selected from Ci-Ce aliphatic and 3- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S.
  • R 4 is oxo
  • R 4 is halogen. In some embodiments, R 4 is flouro.
  • R 4 is -CN.
  • R 4 is -OR a . In some embodiments, R 4 is -OH. In some embodiments, R 4 is -O-Ci-Ce aliphatic (e.g., -OCH3).
  • R 4 is -N(R a ) 2 . In some embodiments, R 4 is -N(H)(R a ). In some embodiments, R 4 is -NH 2 . In some embodiments, R 4 is -N(H)Ci-Ce aliphatic. In some embodiments, R 4 is -N(Ci-Ce aliphatic) 2 .
  • R 4 is -N(H)CH3, - N(H)CH 2 CH 3 , -N(CH 2 CH 3 ) 2J -N(H)CH 2 CH 2 CH 3 , -N(CH 3 ) 2 , -N(CH 3 )CH 2 CH 3 , or - N(CH 3 )CH 2 CH 2 CH3.
  • R 4 is -C(O)R a .
  • R 4 is -C(O)-Ci-C6 aliphatic optionally substituted with -(CH 2 )O ⁇ IOR°.
  • R 4 is -C(O)CH3, - C(O)CH 2 OCH 3 , or -C(O)CH 2 CH 2 OCH 3 .
  • R 4 is -OC(O)R a . In some embodiments, R 4 is -OC(O)-Ci-Ce aliphatic. [0107] In some embodiments, R 4 is -C(O)2R a In some embodiments, R 4 is -C(O)OH. In some embodiments, R 4 is -C(O)2-Ci-Ce aliphatic.
  • R 4 is -C(O)N(R a )2. In some embodiments, R 4 is - C(O)N(H)R a . In some embodiments, R 4 is -C(O)NH 2 . In some embodiments, R 4 is - C(O)N(H)Ci-Ce aliphatic.
  • R 4 is -N(R a )C(O)R a . In some embodiments, R 4 is - N(H)C(O)R a . In some embodiments, R 4 is -N(H)C(O)Ci-Ce aliphatic optionally substituted with -(CH 2 )O-40R°. In some embodiments, R 4 is -N(H)C(O)CH 2 OH or -N(H)C(O)CH 2 OCH 3 .
  • R 4 is an optionally substituted Ci-Ce aliphatic.
  • R 4 is Ci-Ce aliphatic optionally substituted with halogen, -(CH 2 )o-4R°, -(CH 2 )o- 4OR°, -0(CH 2 )o-4R°, -CN, -(CH 2 )O-4N(R°) 2 , or phenyl.
  • R 4 is Ci-Ce aliphatic substituted with halogen, -OH, -OCH3, -CN, or 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
  • R 4 is an optionally substituted 3- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S. In some embodiments, R 4 is optionally substituted 3- to 6-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S. In some embodiments, R 4 is optionally substituted azetidinyl.
  • R 4 is oxo, -OH, -OCH3, fluoro, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, -CN, -CH2F, -CHF 2 , -CH2CHF2, -CH2CH2CHF2, -CH2CH2CF3, - CH 2 CH 2 CH 2 OH, -CH 2 CH 2 CH 2 F, -CH 2 CH 2 CH(CH3) 2 , -CH 2 CH 2 -Ph, -C(CH 3 ) 2 -OH, -CH 2 OCH 3 , 2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2OCH2CH3, -CH2C ⁇ CH, -CH2C ⁇ CHCH3, - CH 2 CN, -CH 2 CH 2 CN, -CH 2 CH 2 CH 2 OCF 3 , -NHCH 2 CH 3 ,
  • each R 4 is independently selected from fluoro, -N(CH3)2, methyl, ethyl, propyl, -CH 2 F, -CHF 2 , -CH 2 CN, and -CH 2 CH 2 CH 2 F. In some embodiments, each R 4 is independently selected from fluoro, methyl, and -CH2F. [0114] In some embodiments, a moiety: [0115] In some embodiments, a moiety: is a moiety selected from:
  • a moiety is a moiety selected from:
  • a moiety is a moiety selected from:
  • a moiety is a moiety selected from:
  • [0121] is a moiety selected from:
  • a moiety is a moiety selected from:
  • a moiety [0123] In some embodiments, a moiety:
  • a moiety is a moiety selected from:
  • R 1 is selected from hydrogen and optionally substituted Ci-Ce aliphatic.
  • R 1 is hydrogen.
  • R 1 is Ci-Ce aliphatic optionally substituted with halogen, -(CH 2 )o-4R°, -(CH 2 )o ⁇ OR°, or -(CH 2 )o ⁇ iPh.
  • R 1 is Ci-Ce aliphatic optionally substituted with halogen or -OH.
  • R 1 is selected from:
  • R 1 is selected from:
  • R 1 is selected from: ,
  • R 2 is selected from hydrogen and optionally substituted
  • R 2 is hydrogen. In some embodiments, R 2 is Ci-Ce aliphatic. In some embodiments, R 2 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments, R 2 is methyl.
  • R 3 is selected from hydrogen, halogen, -CN, -OR a , - C(O)R a , -C(O) 2 R a , -OC(O)R a -C(O)N(R a ) 2 , -OC(O)N(R a ) 2 , -NO 2 , -N(R a ) 2 , -N(R a )C(O)R a , - N(R a )C(O) 2 R a , -N(R a )S(O) 2 R a , -SR a , -S(O) 2 R a , -S(O)N(R a ) 2 , -S(O) 2 N(R a ) 2 , and an optionally substituted Ci-6 aliphatic group.
  • R 3 is hydrogen.
  • n is 0-5. In some embodiments, n is 0. In some embodiments, n is 1, 2, 3, 4, or 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.
  • a compound of Formula I is a compound of Formula II: or a pharmaceutically acceptable salt thereof, wherein A, B, L, n, R 1 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula Il-a:
  • Il-a or a pharmaceutically acceptable salt thereof wherein A, L, R 1 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula Il-b:
  • Il-b or a pharmaceutically acceptable salt thereof wherein A, L, R 1 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula II-c: or a pharmaceutically acceptable salt thereof, wherein A, L, R 1 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula Il-d:
  • a compound of Formula I is a compound of Formula Il-e:
  • Il-e or a pharmaceutically acceptable salt thereof wherein A, L, R 1 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula Il-f: or a pharmaceutically acceptable salt thereof, wherein A, B, L, n, R 1 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula III: or a pharmaceutically acceptable salt thereof, wherein B, L, n, R 1 , R 2 , R 3 , and R 4 are as defined above for Formula T and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula IV: or a pharmaceutically acceptable salt thereof, wherein B, L, n, R 1 , R 2 , R 3 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula V:
  • a compound of Formula I is a compound of Formula VI: or a pharmaceutically acceptable salt thereof, wherein B, L, n, R 1 , R 2 , R 3 , and R 4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • a compound of Formula I is a compound of Formula VII-1 or VII-2:
  • the present disclosure provides compounds selected from Table 1, or a pharmaceutically acceptable salt thereof:
  • the present disclosure provides a compound of Table 2, or a pharmaceutically acceptable salt thereof:
  • the present disclosure provides a compound selected from Table 3, or a pharmaceutically acceptable salt thereof:
  • a compound provided herein is an agonist, and is selected from Table 3.
  • the present disclosure provides a compound selected from Table 4, or a pharmaceutically acceptable salt thereof:
  • a compound provided herein is an antagonist, and is selected from Table
  • the present disclosure provides a compound selected from
  • a compound provided herein is an antagonist, and is selected from Table 5.
  • provided compounds are provided and/or utilized in a salt form (e g., a pharmaceutically acceptable salt form).
  • a salt form e g., a pharmaceutically acceptable salt form.
  • Reference to a compound provided herein is understood to include reference to salts thereof, unless otherwise indicated.
  • Provided compounds may generally be made by the processes described in the ensuing schemes and examples.
  • provided compounds are prepared according to Schemes 1 -4:
  • Scheme 1 [0153] In some embodiments, compounds described herein are prepared via Pictet-Spengler reaction between an appropriately substituted tryptamine (INT 1.1) and a heterocyclic aldehyde of interest (INT 1.2), wherein R 1 , R 2 , R 3 , and A are as defined in classes and subclasses herein with respect to Formula I both singly and in combination, and X is -CN, -CH2OH, -CChMe, - CH2CI, or -CI hBr.
  • INT 1.3 then undergoes selective reduction (when X is -CN), selective oxidation (when X is -CH2OH), or a two-step reduction-oxidation sequence (when X is -CChMe).
  • a B group comprising an amine and substituted with (R 4 )n is coupled with INT 1.4 by reductive amination to provide INT 1.5 (wherein B, R 4 , and n are as defined in classes and subclasses herein with respect to Formula I both singly and in combination).
  • INT 1.3 undergoes a substitution (when X is -CH2CI, or CFFBr) with a B group comprising an amine and substituted with (R 4 ) n to provide INT 1.5 directly.
  • the X group of INT 1.3 is an amine precursor such as -CN or -CH2NHB0C.
  • the intermediates are subjected to reductive amination to provide INT 1.5.
  • target compounds are prepared via a late-stage, two-step conversion of advanced intermediate carboxylic acids to amines.
  • a compound INT 2.1 is saponified to provide carboxylic acid INT 2.2, wherein R 1 , R 2 , R 3 , and A are as defined in classes and subclasses herein with respect to Formula I.
  • INT 2.2 is subjected to amidation with amines of interest (e.g., a B group comprising an amine group and substituted with (R 4 )n), followed by amide reduction to provide compound INT 1.5.
  • preparation of targeted compounds begins with reductive amination between appropriately substituted heterocyclic aldehydes (INT 1.2) and amines of interest (e.g., a B group comprising an amine group and substituted with (R 4 )n), to provide INT 3.1.
  • INT 3.1 undergoes selective reduction (when X is -CN), selective oxidation (when X is -CH2OH), a two-step reduction-oxidation sequence (when X is -CCbMe), or formylation (when X is H) to provide INT 3.2.
  • INT 3.2 is reacted with INT 1.1 in the presence of acid and heat to undergo a Pictet-Spengler reaction to provide INT 1.5.
  • INT 1.3 undergoes a Suzuki -type cross coupling reaction (when X is halogen) with INT 4.1, wherein Y is boronate ester or boronic acid and L is a covalent bond or contains at least one carbon atom (e g., L is -CH2-).
  • INT 4.2 undergoes reduction with LiAlHr to give INT 4.3, wherein R 4 is Me, or acidic deprotection, followed by further functionalization, for example, via reductive amination, SN2 alkylation, or various acylation reactions to give INT 4.3, wherein R 4 is alkyl.
  • provided compounds and compositions are useful in medicine (e.g., as therapy).
  • provided compounds and compositions are useful in research as, for example, analytical tools and/or control compounds in biological assays.
  • the present disclosure provides a composition comprising a compound described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle.
  • the amount of compound in compositions described herein is such that it is effective to measurably induce degradation of a target in a biological sample or in a patient.
  • a composition described herein is formulated for administration to a patient in need of such composition.
  • a composition described herein is formulated for oral administration to a patient.
  • compositions typically contain an active agent (e.g., a compound described herein) in an amount effective to achieve a desired therapeutic effect while avoiding or minimizing adverse side effects.
  • an active agent e.g., a compound described herein
  • provided pharmaceutical compositions comprise a compound described herein and one or more fillers, disintegrants, lubricants, glidants, anti-adherents, and/or anti-statics, etc.
  • Provided pharmaceutical compositions can be in a variety of forms including oral dosage forms, topical creams, topical patches, iontophoresis forms, suppository, nasal spray and/or inhaler, eye drops, intraocular injections forms, deport forms, as well as injectable and infusible solutions. Methods of preparing pharmaceutical compositions are well known in the art.
  • a unit dosage form may be, for example, a liquid pharmaceutical composition containing a predetermined quantity of one or more active agents, a solid pharmaceutical composition (e.g., a tablet, a capsule, or the like) containing a predetermined quantity of one or more active agents, a sustained release formulation containing a predetermined quantity of one or more active agents, or a drug delivery device containing a predetermined amount of one or more active agents, etc.
  • a liquid pharmaceutical composition containing a predetermined quantity of one or more active agents
  • a solid pharmaceutical composition e.g., a tablet, a capsule, or the like
  • sustained release formulation containing a predetermined quantity of one or more active agents
  • a drug delivery device containing a predetermined amount of one or more active agents, etc.
  • compositions may be administered using any amount and any route of administration effective for treating or lessening the severity of any disease or disorder described herein.
  • compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, intraperitoneally, intraci sternally or via an implanted reservoir.
  • the compositions are administered orally, intraperitoneally or intravenously.
  • Sterile injectable forms of the compositions described herein may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3 -butanediol.
  • acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or di-glycerides.
  • Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions.
  • Other commonly used surfactants such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
  • Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
  • delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle.
  • injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactidepolyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
  • provided pharmaceutically acceptable compositions are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions described herein are administered without food. In other embodiments, pharmaceutically acceptable compositions described herein are administered with food. Pharmaceutically acceptable compositions described herein may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fdlers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cety
  • Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
  • the active compounds can also be in micro-encapsulated form with one or more excipients as noted above.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art.
  • the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch.
  • Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose.
  • the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.
  • Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • compositions described herein may be administered in the form of suppositories for rectal administration.
  • suppositories for rectal administration.
  • suppositories can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug.
  • suitable non-irritating excipient include cocoa butter, beeswax and polyethylene glycols.
  • compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • compositions described herein may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
  • Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.
  • compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers.
  • Carriers for topical administration of compounds described herein include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.
  • provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers.
  • Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
  • compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride.
  • the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.
  • compositions described herein may also be administered by nasal aerosol or inhalation.
  • Such compositions are prepared according to techniques well- known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
  • Dosage forms for topical or transdermal administration of a compound disclosed herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches.
  • the active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required.
  • Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this disclosure.
  • the present disclosure contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body.
  • Such dosage forms can be made by dissolving or dispensing the compound in the proper medium.
  • Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
  • the estrogen receptor (“ER”) is involved in a variety of biological processes, relating, for example, to development of the female reproductive system, maintenance of bone mass, protection of cardiovascular and/or central nervous system components, etc. (see, for example, Pearce & Jordan Crit. Rev. Onc/Hem 50:3, 2004; Heldring Phys. Rev. 87:905, 2007).
  • the ER has been implicated in a variety of cancers. In many tumors that express the estrogen receptor (i.e., ER + tumors), active ERa signaling has been demonstrated to drive cell proliferation (although ERp signaling has been reported to be able to achieve tumor suppressor effects; see, for example, Nilsson & Gustafson Clin. Pharmacol. Ther. 89:44, 2011).
  • tumors e.g., breast tumors
  • therapies targeting the ER are standard of care for many patients with ER + tumors (see, for example, Cardoso et al Annals One. ⁇ https://doi.org/10.1093/announc/mdmx036>, 2017; Rugo et al. J. Clin. Oncol. 34:3069, 2016; Senkus et al Annal One. 26:v8, 2015; Sareddy & Vadlamudi Clin. J Nat. Med, 13:801, 2015).
  • recommended therapy typically involves tumor resection, followed by ER-targeted therapy (e.g., as discussed below).
  • ER- targeted therapy is the mainstay.
  • some ER-targeting agents are designed and/or documented to reduce levels of estrogen (i.e., 17(3 estradiol) production. In other embodiments, some ER-targeting agents are designed and/or documented to increase levels of estrogen production.
  • ER-targeting agents are designed and/or documented to bind directly to the ER; in some cases, such agents compete with estrogen for binding to the ER and/or interfere with the allosteric changes that estrogen binding would naturally produce. Often, the term “antiestrogen” is used to refer to agents that bind to the ER, and sometimes is specifically used to indicate those agents that compete with estrogen for ER binding.
  • SERM selective estrogen receptor modulator
  • compounds provided herein are estrogen receptor antagonists.
  • an “estrogen receptor antagonist” refers to a compound or composition that produces an antagonistic effect when contacting the estrogen receptor of a subject or biological sample.
  • an estrogen receptor antagonist is characterized by having:
  • compounds provided herein are complete estrogen receptor antagonists.
  • a complete estrogen receptor antagonist (a “CERAN”) is one that (1) inhibits both AF1 and AF2, and in particular inhibits AF1 activity that remains present in constitutively active ER mutants; (2) promotes ER degradation; and (3) lacks the partial ER agonist activity observed with certain other agents.
  • CERAN complete estrogen receptor antagonist
  • many previous therapies including for example, ARN-810, AZD9496, tamoxifen, and others, are less effective than CERANs at least in part because they only partially antagonize ER, and specifically because they inhibit activation of AF2 but not AF1.
  • an estrogen receptor antagonist is characterized by having (i) a pICso greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode) of Example 209.
  • an estrogen receptor antagonist is characterized by having (i) a pICso greater than 7.5 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode) of Example 209.
  • compounds provided herein are estrogen receptor agonists.
  • an “estrogen receptor agonist” refers to a compound or composition that produces an agonistic effect when contacting the estrogen receptor of a subject or biological sample.
  • an estrogen receptor agonist is characterized by having (i) at least 80% increase in the E2 -normalized signal in the AP assay (agonist mode) of Example 209 and (ii) no more than 80% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209.
  • a disease, disorder, or condition is a cancer.
  • a disease, disorder, or condition is associated with a mutation in an estrogen receptor.
  • provided compounds are useful for treating a disorder associated with increased ER activity (e.g., an ER-associated cancer such as breast cancer).
  • provided estrogen receptor antagonists e.g., complete estrogen receptor antagonists
  • provided compounds are useful for treating a disorder associated with decreased ER activity (e.g., menopause-related conditions or symptoms, or osteoporosis).
  • provided estrogen receptor agonists are useful for treating such disorders.
  • the present disclosure provides a method of treating a disorder mediated by an estrogen receptor in a subject, comprising administering to the subject a therapeutically effective amount of a compound described herein.
  • a disorder is selected from the group consisting of breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, uterine cancer, and endometriosis.
  • a disorder is breast cancer.
  • a disorder is ovarian cancer.
  • a disorder is endometrial cancer.
  • a disorder is vaginal cancer.
  • a disorder is lung cancer.
  • a disorder is bone cancer.
  • a disorder is uterine cancer.
  • a disorder is endometriosis.
  • the present disclosure provides a method of treating a disorder associated with a mutation of an estrogen receptor in a subject, comprising administering to the subject a therapeutically effective amount of a compound described herein.
  • a disorder is selected from the group consisting of breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, uterine cancer, and endometriosis.
  • a disorder is breast cancer.
  • a disorder is ovarian cancer.
  • a disorder is endometrial cancer.
  • a disorder is vaginal cancer.
  • a disorder is lung cancer.
  • a disorder is bone cancer.
  • a disorder is uterine cancer.
  • a disorder is endometriosis.
  • a method of treating a disorder in a subject described herein comprises administering to the subject a compound described herein in combination with or alternation with an anti-cancer agent.
  • an anti-cancer agent is selected from an mTOR inhibitor, a CDK4/6 inhibitor, a PI3 kinase inhibitor, an aromatase inhibitor, an antibody to or inhibitor of PD-1, PD-L1 or CTLA-4, or an antibody to or inhibitor of EGFR, PGFR, or IGFR.
  • an anti-cancer agent is a HER2 inhibitor.
  • a HER2 inhibitor is selected from tucatinib, trastuzumab, pertuzumab, ado- trastuzumab, trastuzumab emtansine, ado-trastuzumab emtansine, trastuzumab deruxtecan pertuzumab, lapatinib, and neratinib.
  • an anti-cancer agent is an mTOR inhibitor.
  • an mTOR inhibitor is selected from everolimus sirolimus, temsirolimus, and LY3023414.
  • an anti-cancer agent is a CDK4/6 inhibitor.
  • a CDK4/6 inhibitor is selected from palbociclib, abemaciclib, ribociclib, lerociclib, trilaciclib, and SHR6390.
  • an anti-cancer agent is a PI3 kinase inhibitor.
  • a PI3 kinase inhibitor is selected from perifosine, CAL101, BEZ235, XL147, XL765, GDC-0941, and IPI-145.
  • a PI3 kinase inhibitor is a PIK3CA inhibitor.
  • a PIK3CA inhibitor is selected from alpelisib, taselisib, and LY3023414.
  • an anti-cancer agent is an aromatase inhibitor.
  • an aromatase inhibitor is selected from aminoglutethimide, testolactone, anastrozole, letrozole, exemestane, vorozole, formestane, fadrozole, 4-hydroxyandrostenedione, 1, 4, 6-androstatrien-3, 17-dione, and 4-androstene-3, 6, 17-trione.
  • an anti-cancer agent is an antibody to or inhibitor of PD-1, PD-L1 or CTLA-4.
  • an anti-cancer agent is an antibody to or inhibitor of EGFR, PGFR, or IGFR. In some embodiments, an anti-cancer agent is erlotinib or gefitinib.
  • a method described herein comprises administering a compound reported herein in combination or in alternation with an estrogen receptor antagonist or a partial estrogen receptor antagonist.
  • the present disclosure provides a method of preventing recurrence of a cancer in a subject comprising administering to the subject a compound described herein.
  • a cancer is selected from breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, and uterine cancer.
  • a compound described herein is administered as an adjunctive therapy after or instead of chemotherapy, radiation, or surgery.
  • a compound is administered after surgery.
  • a compound is administered prior to surgery.
  • a cancer is a breast cancer that has progressed in the presence of endocrine or aromatase therapy.
  • Example 1 Methyl 3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylate.
  • Example 2 3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylic acid.
  • Example 3 (3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indol-6-yl)methanol.
  • the reaction mixture was cooled to 0 °C, 0.06 mL H2O was added dropwise to the reaction mixture, followed by 0.12 mL 10% NaOH solution, then 0.24 mL H2O, and stirred at room temperature for 30 min. The mixture was then fdtered and concentrated under vacuum to afford crude product.
  • the crude product was purified by normal phase Combiflash chromatography (0% - 60% ethyl acetate in hexane) to afford the title compound (268.3 mg, 95.6% yield) as paleyellow solid.
  • Example 4 3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde.
  • Example 7 3-Fluoro-N-((3-((l R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4, 9-tetrahydro- lH-pyrido[3,4-b]indol-l-yl)-lH-indol-6-yl)methyl)-N-methylpropan-l-amine
  • Example 8 N-((3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH indol-6-yl)methyl)-N-methylpropan-l-amine.
  • Example 9 3-Fluoro-N-((3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4, 9-tetrahydro- lH-pyrido[3,4-b]indol-l-yl)-lH-indol-6-yl)methyl)propan-l-amine
  • Example 10 Methyl 3- ((JS,3R)-2- (2- Fluor o- 2- methylpropyl)-3 -methyl- 2, 3, 4, 9 -tetrahydro- 1 H- pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carboxylate.
  • Example 11 3-((lS,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carboxylic acid.
  • Example 12 (3-((lS,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3, 4-b]indol-l -yl)-lH-indazol- 6-yl) methanol.
  • the reaction mixture was cooled to 0 °C, 0.12 mL H2O was added dropwise to the reaction mixture, then added 0.24 mL of 10% NaOH solution, followed by 0.48 mL of H2O, stirred at room temperature for 30 min, then filtered and concentrated under vacuum to afford crude product.
  • the crude product was purified by normal phase Combiflash chromatography (0%-60% ethyl acetate in hexane) to afford the title compound (350.00 mg, 68.0% yield) as pale-yellow solid.
  • Example 13 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde.
  • Example 14 (lS,3R)-2-(2-Fluoro-2-methylpropyl)-l-(6-((3-(fluoromethyl)azetidin-l- yl)niethyl)-lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 15 (lS,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-l-(6-((3-methylazetidin-l- yl)methyl)-lH-indazol-3-yl)-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 16 (1 S,3R)-1 -(6-((3-ethylazetidin-l -yl)methyl)-lH-indazol-3-yl)-2-(2-fluoro-2- methyl-propyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 17 3-Fluoro-N-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9- tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazol-6-yl)methyl)-N-methylpropan-l-amine.
  • Example 18 N-((3-((l S, 3R)-2- (2-Fluoro-2-methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH indazol-6-yl)methyl)-N-methylpropan-l-amine.
  • Example 19 3-Fluoro-N-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2, 3,4,9- tetrahydro- 1 H-pyrido[3,4-b]indol-l -yl)-l H-indazol-6-yl)methyl)propan-l -amine.
  • Example 20 (lS,3R)-2-(2-fluoro-2-methylpropyl)-l-(6-(((R)-3-fluoropyrrolidin-l-yl)methyl)- lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 21 (lS,3R)-2-(2-fluoro-2-methylpropyl)-l-(6-(((S)-3-fluoropyrrolidin-l-yl)methyl)- lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 22 (lS,3R)-l-(6-(((R)-3-ethylpyrrolidin-l-yl)methyl)-lH-indazol-3-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH-pyrido[3, 4-b]indole.
  • Example 23 (lS,3R)-l-(6-(((S)-3-ethylpyrrolidin-l-yl)methyl)-lH-indazol-3-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH-pyrido[3, 4-b]indole.
  • Example 24 (lS,3R)-l-(6-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)-lH-indazol-3-yl)-2-(2- fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 25 (1 S,3R)-2-(2-fluoro-2-methylpropyl)-l -(6-(((R)-3-(fluoromethyl)pyrrolidin-l - yl)methyl)-lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 26 (lS,3R)-2-(2-fluoro-2-methylpropyl)-l-(6-(((S)-3-(fluoromethyl)pyrrolidin-l- yl)methyl)-lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 29 6-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH pyrido-[3,4-b]indol-l-yl)-lH-indazol-6-yl)methyl)-2-oxa-6-azaspiro[3.3]heptane.
  • Example 30 2-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]-indol-l-yl)-lH-indazol-6-yl)methyl)-2-azaspiro[3.3]heptan-6-ol.
  • Example 31 l-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-LE[- pyrido-[3,4-b]indol-l-yl)-lE[-indazol-6-yl)methyl)azetidine-3-carbonitrile.
  • Example 32 (lS,3R)-l-(6-((6-fluoro-2-azaspiro[3.3]heptan-2-yl)methyl)-lH-indazol-3-yl)-2- (2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
  • Example 33 (1S,3R)-1-(6-((6,6-difluoro-1-azaspiro[3.3]heptan-1-yl)methyl)-1H-indazol-3-yl)- 2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole.
  • Example 34 Methyl 3-((lR,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylate.
  • Example 35 (3-((lR,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lEl-pyrido[3,4-b]indol-l-yl)-lE[-indol-6-yl)methanol.
  • Example 36 3-((lR,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde.
  • Example 39 (3-((lS,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazol-6-yl)methanol.
  • Example 40 3-((lS,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde.
  • Example 41 2,2-difluoro-3-((lS,3R)-l-(6-((3-(fluoromethyl)azetidin-l-yl)methyl)-lH-indazol- 3-yl)-3-methyl-l,3,4,9-tetrahydro-2H-pyrido[3,4-b]indol-2-yl)propan-l-ol.
  • Example 48 Methyl 2-[(3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-5-carboxylate.
  • Example 49 [2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-5-yl]methanol.
  • Example 50 2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-5-carbaldehyde.
  • Example 55 2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lE[-indole-6-carbaldehyde.
  • the reaction was the quenched with ethyl acetate, warmed to 0 °C and then treated with saturated solution of Rochelle's salt. The reaction mixture was stirred for 30 mins, and the layers separated. The aqueous phase was extracted with EtOAc, washed with brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-30% EtOAc in hexane to give title compound (60 mg, 0.149 mmol, 56.72 % yield). 404.2 [M+H] + .
  • Example 56 (lR,3R)-l-[6-[[3-(fluorometbyl)azetidin-l-yl]metliyl]-lH-indol-2-yl]-2-(2-fluoro- 2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • Example 61 (lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l-[2-[(3-methylazetidin-l- yl)methyl]-lH-indol-5-yl]-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • Example 64 (lR,3R)-2-(2-fluoro-2-methyl-propyl)-l-[2-[[(3S)-3-fluoropyrrolidin-l- yl]methyl]-lH-indol-5-yl]-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • LCMS: m/z 477.3 [M+H] + .
  • Example 65 [5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methanol.
  • Example 66 5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde.
  • Example 68 2,2-difluoro-3-[(lR,3R)-3-methyl-l-[2-[(3-methylazetidin-l-yl)methyl]-lfI-indol- 5-yl]-l, 3, 4, 9-tetrahydropyrido[3, 4-b]indol-2-yl]propan-l -ol.
  • Example 70 [6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-2-yl]methanol.
  • Example 71 6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde.
  • reaction mixture was stirred for 1 h and treated with sodium triacetoxyborohydride (3.00 equiv, 39 mg, 0.186 mmol), and then stirred for additional 2 h.
  • the reaction was quenched with 1 mL of methanol and purified by C-18 reverse phase chromatography using a Phenomenex Luna column, eluting with 10-50% ACN in water with 0.1% formic acid as modifier, to give title compound (4.5 mg, 0.00944 mmol, 15.24 % yield). 477.2 [M+H] + .
  • Example 73 Methyl 5-[(lR, 3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3, 4-b]indol-l -yl]benzofuran-2-carboxylate.
  • Example 75 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]benzofuran-2-carbaldehyde.
  • Example 78 Methyl 3-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-5-carboxylate.
  • Example 80 3-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l-yl]-lH-indazole-5-carbaldehyde.
  • Example 81 (1 R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl- 1 -[5-[(3-methylazetidin- 1 - yl)methyl]-lH-pyrrolo[2,3-b]pyridin-3-yl]-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • Example 82 (lR,3R)-l-[5-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-pyrrolo[2,3-b]pyridin-3- yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • Example 83 (lR,3R)-l-[5-(3-azabicyclo[3.1.0]hexan-3-ylmethyl)-lEI-pyrrolo[2,3-b]pyridin-3- yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • Example 84 Methyl 2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-6H-thieno[2,3-b]pyrrole-5-carboxylate.
  • 2-Fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l -amine 550 mg, 2.2 mmol
  • DCE 13 mL
  • Example 86 2-[(lS,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-6H-thieno[2,3-b]pyrrole-5-carbaldehyde.
  • Example 87 2-[(lS,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- bJindol-l-ylJ-5-[(3-methylazetidin-l-yl)methylJ-6H-thieno[2,3-bJpyrrole.
  • Example 88 5-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-2-[(lS,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-6H-thieno[2,3-b]pyrrole.
  • Example 89 Methyl 2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole-5-carboxylate.
  • Example 90 2-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l -yl]-4H-thieno[3,2-b]pyrrole-5-carboxylic acid.
  • Example 91 [2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-4H-thieno[3,2-b]pyrrol-5-yl]methanol.
  • Example 92 2-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l-yl]-4H-thieno[3,2-b]pyrrole-5-carbaldehyde.
  • Example 93 5-[[3-(fluoromethyl)azetidin-l-yl]methyl]-2-[(lS,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole.
  • Example 94 2-[( IS, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl- 1, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l-yl]-5-[(3-methylazetidin-l-yl)methyl]-4H-thieno[3,2-b]pyrrole.
  • Example 95 3-fluoro-N-[[2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrol-5-yl]methyl]propan-l-amine.
  • Example 96 5-[[(3S)-3-ethylpyrrolidin-l-yl]methyl]-2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole.
  • Example 97 Methyl 5-bromo-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-2- carboxylate.
  • Example 99 2-(Hydroxymethyl)-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-bJpyridine-5- carbaldehyde.
  • Example 100 [5-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-2- yljmethanol.
  • Example 101 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-2-carbaldehyde.
  • Example 102 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-2-carbaldehyde.
  • Example 103 (lR,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-pyrrolo[2,3-b]pyridin- 5-yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • Example 104 Ethyl 5-chloro-l-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridine-2- carboxylate.
  • Example 107 Ethyl 5-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-b]pyridine-2- carboxylate
  • Example 108 (5-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lEI- pyrido[3,4-b]indol-l-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-b]pyridin-2- yl) methanol
  • Example 110 5-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4-
  • Example 111 (lS,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-pyrrolo[3,2-b]pyridin- [0321]
  • 3-(fluoromethyl)azetidine hydrochloride (2.00 equiv, 28 mg, 0.223 mmol) in NMP (1 mL) was added the triethylamine (2.00 equiv, 0.031 mL, 0.223 mmol), stirred for 5 min, followed by addition of 5-[(lS,3R)-2-(2-fhioro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-pyrrolo[3,2-b]pyridine-2-carbaldehyde (1.00 equiv, 45 mg, 0.111 mmol) and acetic acid (2.00 equiv, 0.0
  • Example 112 2-[(3-methylazetidin-l-yl)methylJ-3l l-benzimidazole-5-carbonitrile.
  • Example 114 (lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l-[2-[(3-methylazetidin-l- yl) methyl]-3H-benzimidazol-5-yl]-l,3, 4, 9-tetrahydropyrido[3 , 4-b/indole.
  • Example 116 [2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazol-6-yl]methanol.
  • Example 118 2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-6-[(lR,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l, 3, 4, 9-tetrahydropyrido[3, 4-b]indol-l -yl]-l, 3-benzoxazole.
  • Example 119 Ethyl 6-bromoimidazo[l,2-a]pyridine-2-carboxylate.
  • Ethyl 6-bromoimidazo[l,2-a]pyridine-2-carboxylate (4.6 g, 17.1 mmol) was dissolved in THF (20 mL) under nitrogen and brought to 0 °C. DIBAL (34 mmol) was added and stirred for 1.5 h. The reaction was brought to ambient temperature and saturated Rochelle's Salt solution was added and stirred for 30 minutes. The aqueous layer was separated and extracted with EtOAc twice, and the organic layers were combined then washed with saturated brine solution. It was then dried over sodium sulfate, filtered, and dried in vacuo.
  • Example 122 2-[[Tert-butyl(dimethyl)silyl]oxymethyl]imidazo[l,2-a]pyridine-6-carbaldehyde.
  • Example 123 Tert-butyl-[[6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]imidazo[l,2-a]pyridin-2-yl]methoxy]-dimethyl-sdane.
  • Example 124 [6-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4, 9- tetrahydropyrido[3,4-b]indol-l-yl]inddazo[l,2-a]pyridin-2-yl]niethanoL
  • Example 125 6-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4, 9- tetrahydropyrido[3,4-b]indol-l-yl]imidazo[l,2-a]pyridine-2-carbaldehyde.
  • Example 126 (lR,3R)-l-[2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]imidazo[l,2-a]pyridin-6- yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
  • Example 127 2- (hydroxyniethyl)inudazo[l, 2-a]pyridine- 7-carbonitrile [0338] To 7-cyano-imidazo[l,2-a]pyridine-2-carboxylic acid ethyl ester (1.00 equiv, 250 mg, 1.16 mmol) in THF (10 mL) at 0 °C under nitrogen was added diisobutylaluminum hydride (2.00 equiv, 2.3 mL, 2.32 mmol) dropwise and stirred for 1 h. After slowly warming to room temp, the reaction was cooled back to 0 °C, and quenched with EtOAc, followed by saturated solution of Rochelle's salt. The mixture was stirred for 30 min.
  • Example 135 2-(chloromethyl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
  • Example 137 2-[[3-methylazetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3- methyl-1, 3, 4, 9-tetrahydropyrido[3,4-b]indol-l-yl]-l, 3-benzoxazole.
  • Example 138 2-(2-azaspiro[3.3]heptan-2-ylmethyl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)- 3-methyl-l ,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
  • Example 139 5-Bromo-2-(chloromethyl)-6-fluoro-l,3-benzoxazole.
  • 2 -Amino-4-bromo-5-fhiorophenol (1.00 equiv, 640 mg, 3.11 mmol) and ethyl 2- chloroethanimidoate hydrochloride (1.30 equiv, 638 mg, 4.04 mmol) were heated at 90 °C in ethanol (10 mL) overnight. The volatiles were removed under reduced pressure, and the crude was dissolved in DCM. The insoluble material was filtered, and the solution was concentrated under reduced pressure to give the title product (783 mg, 95% yield).
  • LCMS: m/z 265.5 [M+H] + .
  • Example 140 5-Bromo-6-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole.
  • Example 141 6-Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5- carbonitrile.
  • Example 143 6-Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2- methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
  • the solution was diluted with DCM, poured into saturated sodium bicarbonate solution, and the organic layers were separated. The organic layers were dried over sodium sulfate, filtered, and dried in vacuo.
  • the crude was dissolved in DMF (3 mL) and was purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (7.7 mg, 7% yield).
  • Example 144 5-Bromo-2-(chloromethyl)-7-fluoro-l,3-benzoxazole.
  • Example 145 5-Bromo-7-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole.
  • Example 147 7-Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5- carbaldehyde.
  • Example 148 7-Eluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2- methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
  • the solution was diluted with DCM, poured into saturated sodium bicarbonate solution, and the organic layers were separated. The organic layers were dried over sodium sulfate, filtered, and dried in vacuo.
  • the crude was dissolved in DMF (2 mL) and was directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (115.9 mg, 55% yield).
  • Example 149 Tert-butyl-[3-[(lR,3R)-l-[2-(chloromethyl)-l,3-benzoxazol-5-yl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]-2,2-difluoro-propoxy]-diphenyl-silane.
  • Example 150 3-[(lR,3R)-l-[2-(Chloromethyl)-l,3-benzoxazol-5-yl]-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-2-yl]-2,2-difluoro-propan-l-ol.
  • Example 151 2,2-Difluoro-3-[(lR,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3- benzoxazol-5-yl]-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-l-ol.
  • reaction was quenched with formic acid (80 pL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water), then lyophilized to give the title compound as a white solid (28.7 mg, 29% yield).
  • Example 152 Tert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[7-fluoro-2-[[3-(fluoromethyl)azetidin-l- yl]methyl]-l ,3-benzoxazol-5-yl]-3-methyl-l ,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propoxy]- diphenyl-silane.
  • Example 153 2,2-Difluoro-3-[(lR,3R)-l-[7-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl] ⁇ l,3-benzoxazol-5-yl]-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-L-ol.
  • Example 155 Tert-butyl 3-(5-bromo-l,3-benzoxazol-2-yl)azetidine-l-carboxylate.
  • Example 156 Tert-butyl 3-(5-cyano-l,3-benzoxazol-2-yl)azetidine-l-carboxylate.
  • Tert-butyl 3-(5-bromo-l,3-benzoxazol-2-yl)azetidine-l-carboxylate (1.00 equiv, 536 mg, 1.52 mmol)
  • tetrakis(triphenylphosphine)palladium(0) 0.0500 equiv, 88 mg, 0.0759 mmol
  • zinc cyanide (1.00 equiv, 178 mg, 1.52 mmol) were dissolved in dry DMF and under an atmosphere of nitrogen and stirred at 80 °C overnight.
  • Example 157 Tert-butyl 3-(5-formyl-l,3-benzoxazol-2-yl)azetidine-l-carboxylate.
  • Example 158 Tert-butyl 3-[5-[(lR,3R)-2-(2-fluoro-2-metliyl-propyl)-3-metliyl-l,3,4,9- tetrahydropyrido[3, 4-b]indol-l-yl]-l, 3-b enzoxazol-2-yl] azetidine- 1-carboxy late.
  • Example 159 2-(Azetidin-3-yl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
  • reaction was directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water), then lyophilized to give the title compound as a white solid (6.1 mg, 28% yield).
  • Example 161 2-[l-[[5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]azetidin-3-yl]propan-2-ol.
  • Example 162 6-(Difluoromethyl)-2-[[5-[(lR)-2-(2-fluoro-2-metliyl-propyl)-3,3-dimetliyl-4,9- dihydro-lH-pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]-2-azaspiro[3.3]heptan-6-ol.
  • 6-(difluoromethyl)-2- azaspiro[3.3]heptan-6-ol hydrochloride 10 mg, 0.05 mmol was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size.
  • Example 163 l-[[5-[(lR)-2-(2-Fluoro-2-tnethyl-propyl)-3,3-dimethyl-4,9-diliydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]azetidine-3-carbonitrile.
  • Example 164 (lR)-2-(2-Fluoro-2-methyl-propyl)-l-[2-[[3-(methoxymethyl)azetidin-l- yl]methyl]-lH-indol-5-yl]-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4-b]indole.
  • Example 165 (lR)-l-[2-[(6-Fluoro-2-azaspiro[3.3]heptan-2-yl)metliyl]-lH-indol-5-yl]-2-(2- fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4-b]indole.
  • 6-fluoro-2- azoniaspiro[3.3]heptane;2,2,2-trifluoroacetate 11 mg, 0.05 mmol was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours.
  • Example 166 2-[[5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]-2-azaspiro[3.3]heptan-6-oL
  • Example 167 6-[[5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]-2-oxa-6-azaspiro[3.3]heptane.
  • the residue was purified on a Biotage automated chromatography system (11 g, Biotage 50 pm KP-amino-d column), eluting with a gradient of 0 to 100% ethyl acetate in hexanes to give the title compound (50 mg, -85% purity).
  • the resulting residue was loaded onto Celite® (1 g) purified on a Biotage automated chromatography system (5.5 g Redigold C18 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound (21 mg, 31% yield) a pale-yellow solid.
  • Example 171 2-(l-((5-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indol-2-yl)methyl)azetidin-3-yl)acetonitrile
  • Example 173 (lR,3R)-l-(2-((3-(Difluoromethyl)azetidin-l-yl)methyl)-lH-indol-5-yl)-2-(2- fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole
  • Example 174 l-((5-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indol-2-yl)methyl)-N,N-dimethylazetidin-3-amine
  • Example 175 (1R,3R)-1-(2-((3,3-Dimethylazetidin-1-yl)methyl)-1H-indol-5-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole [0386] A solution of 3,3 (37 mg, 0.297 mmol, 2 equiv) and triethylamine (41 ⁇ L, 0.297 mmol, 2 equiv) in dichloromethane (2 mL) was added to a solution of 5-[(1R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-1,3,4,9-tetrahydropyrido[3,4- b]indol-1-yl]-1H-indole-2-carbaldehyde (60 mg, 0.149 mmol, 1 equiv) in dichlorome
  • Example 176 (1R,3R)-2-(2-Fluoro-2-methylpropyl)-1-(2-((3-methoxy-3-methylazetidin-1- yl)methyl)-1H-indol-5-yl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole [0387]
  • a solution of 3-methoxy-3-methylazetidine hydrochloride (41 mg, 0.297 mmol, 2 equiv) and triethylamine (41 ⁇ L, 0.297 mmol, 2 equiv) in dichloromethane (2 mL) was added to a solution of 5-[(1R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-1,3,4,9-tetrahydropyrido[3,4- b]indol-1-yl]-1H-indole-2-carbaldehyde (
  • Example 177 1R,3R)-1-(2-((3,3-Difluoroazetidin-1-yl)methyl)-1H-indol-5-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole 9 mg, 0.297 mmol, 2 equiv) and triethylamine (41 ⁇ L, 0.297 mmol, 2 equiv) in dichloromethane (2 mL) was added to a solution of 5-[(1R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-1,3,4,9-tetrahydropyrido[3,4- b]indol-1-yl]-1H-indole-2-carbaldehyde (60 mg, 0.149 mmol, 1 equiv) in dichloromethane (2 mL).
  • Example 178 (lR,3R)-2-(2-Fluoro-2-methylpropyl)-l-(2-((3-fluoro-3-methylazetidin-l- yl)methyl)-lH-indol-5-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole
  • Example 179 (5-((lR,3R)-3-Methyl-2-(2,2,2-trifluoroethyl)-2,3,4,9-tetrahydro-lH-pyrido[3,4- b]indol-l-yl)-lH-indol-2-yl)methanol
  • Example 180 5-((lR,3R)-3-Methyl-2-(2,2,2-trifluoroethyl)-2,3,4, 9-tetrahydro-lH-pyrido[3,4- b]indol-l-yl)-lH-indole-2-carbaldehyde
  • Example 181 (1R,3R)-1-(2-((3-(Fluoromethyl)azetidin-1-yl)methyl)-1H-indol-5-yl)-3-methyl- 2-(2,2,2-trifluoroethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole [0392] A mixture of roethyl)-2,3,4,9-tetrahydro-1H- pyrido[3,4-b]indol-1-yl)-1H-indole-2-carbaldehyde (40 mg, 0.1 mmol, 1.0 equiv), 3- (fluoromethyl)azetidine hydrochloride (24 mg, 0.2 mmol, 2.0 equiv), 1.0 M triethylamine in dichloromethane (0.19 mL, 0.19 mmol, 2.0 equiv), and 1.0 M acetic acid in dichloromethan
  • Example 184 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole-5-carbonitrile.
  • Example 185 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole-5-carbaldehyde.
  • Example 186 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l, 3, 4, 9-tetrahydropyrido[3, 4-b]indol-l -yl]-l, 3-benzothiazole.
  • Example 188 tert-Rutyl 3-(5-formylindazol-l-yl)azetidine-l -carboxylate and tert-butyl 3-(5- formylindazol-2-yl)azetidine-l-carboxylate
  • the organic layer was washed with 5% LiCl (50 mL x 2).
  • the organic layers were dried over sodium sulfate and purified via silica gel (40 g) eluting with 25% hexanes:EtOAc to afford tert-butyl 3-(5-formylindazol-l-yl)azetidine-l-carboxylate as a beige solid (844 mg, 2.80 mmol, 51.4% yield) and with 35% hexanes:EtOAc to afford tert-butyl 3-(5-formylindazol-2- yl)azetidine-l -carboxylate as a beige solid (514 mg, 1.71 mmol, 31.3 % yield).
  • Example 189 tert-Butyl 3-[5-[(l R,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]- [0402]
  • a solution of (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)- 2,2-difluoropropan-l -amine (1.20 equiv, 456 mg, 0.900 mmol)
  • tert-butyl 3-(5-formylindazol-l- yl)azetidine-l -carboxylate (1.00 equiv, 226 mg, 0.750 mmol)
  • acetic acid (3.00 equiv, 0.13 mL, 2.25 mmol) in DCE (1.5 mL) was heated at 125 °C for 3 h.
  • Example 190 tert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]-3-methyl-
  • Example 191 2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]-3-methyl-l,3,4,9- [0404] To a solution oftert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propoxy]-diphenyl-silane;2,2,2- trifluoroacetic acid (1.00 equiv, 52 mg, 0.0566 mmol) in THF (0.5 mb) was added tetra-n- butylammonium fluoride (5.30 equiv, 0.30 mL, 0.300 mmol).
  • Example 194 tert-butyl 3-[5-[(l R,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]indazol-2-yl]azetidine-l-carboxylate
  • Example 200 2-(Hydroxymethyl)-l-(p-tolylsulfonyl)indole-5-carbaldehyde.
  • Example 201 [5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(p-tolylsulfonyl)indol-2-yl]methanol
  • Example 202 [5-[(lR,3R)-3-methyl-2-(2, 2,3,3, 3-pentafluoropropyl)-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l-(p-tolylsulfonyl)indol-2-yl]methanol
  • Example 203 [5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9- tetrahydropyrido[3, 4-b]indol-l -yl]-lH-indol-2-yl]methanol
  • Example 204 [5-[(lR,3R)-3-Methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4, 9- tetrahydropyrido[3, 4-b]indol-l -yl]-LH-indol-2-yl] methanol
  • Example 205 5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde
  • Example 206 5-[(lR,3R)-3-Methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde
  • 3- (fluoromethyl)azetidine hydrochloride (1.13 equiv, 36 mg, 0.18 mmol) was dissolved in DMF (1 mL) and TEA (2.2 equiv, 50 pL, 0.35 mmol) was added. These solutions were combined and stirred for 30 min. STAB (3.0 equiv, 104 mg, 0.48 mmol) was added and stirred for 1 h.
  • reaction was quenched with MeOH (1 mL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-45% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (36.7 mg, 0.07 mmol, 44% yield).
  • Example 208 (1R,3R)-1 -[2-[[3- (Fluoromethyl) azetidin- 1 -yljmethyl]- 1 H-indol-5-yl]-3- methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4-b]indole
  • 3- (fluoromethyl)azetidine hydrochloride (1.08 equiv, 17 mg, 0.13 mmol) was dissolved in DMF (1 mL) and TEA (2.2 equiv, 37 pL, 0.26 mmol) was added. These solutions were combined and stirred for 30 min. STAB (3.00 equiv, 77.2 mg, 0.36 mmol) was added and stirred for 1 h.
  • reaction was quenched with MeOH (1 mL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-45% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (29.8 mg, 0 056 mmol, 46% yield).
  • ECC-1 cells were trypsinized and resuspended in hormone-depleted media and plated at a density of 15 k cells per well into a 96-well plate for at least 4 hours. Cells were treated with test compounds in the absence of E2 (for agonist mode) or in the presence of 500 pM E2 (for antagonist mode) for 3 days and plates were subsequently frozen at -80 °C. Thawed plates were incubated with a chromogenic substrate of AP, p-nitrophenyl phosphate (Thermo Fisher Scientific), for 40 minutes at 42 °C, and absorbances were read at 405 nm. AP activity was normalized to the activity of 500 pM E2 alone. This assay was shown to correlate with the in vivo studies comparing uterine wet weight in ovariectomized rats following treatment with a number of antiestrogens.
  • Table 6 shows estrogen receptor modulation (e.g., agonism and antagonism) of certain compounds of the present disclosure.
  • the compound numbers correspond to the compound numbers of Table 1 and 2.
  • Compounds having an activity designated as “+” provided estrogen receptor agonism having (i) at least 80% increase in the E2-normalized signal in the AP assay (agonist mode) and (ii) no more than 80% reduction in the E2-normalized signal in the AP assay (antagonist mode).
  • Compounds having an activity designated as “++” provided estrogen receptor antagonism having (i) between 10% and 80% increase in the E2 -normalized signal in the AP assay (agonist mode) and (ii) between 10% and 80% reduction in the E2 -normalized signal in the AP assay (antagonist mode).
  • Compounds having an activity designated as “+++” provided estrogen receptor antagonism (i) with pICso greater than 7.5 and at least a 10% reduction in the E2 -normalized signal in the AP assay (antagonist mode); and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode).
  • particularly useful modulators of the estrogen receptor are compounds having greater than “+” activity in Table 6. In some embodiments, particularly useful modulators of the estrogen receptor are compounds having greater than “++” activity in Table 6. In some embodiments, particularly useful modulators of the estrogen receptor are compounds having “+++” activity in Table 6.
  • Compound 1-109 and Compound 1-110 provided estrogen receptor antagonism with (i) pICso greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode); and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode).

Abstract

The present disclosure provides 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole derivatives of formula I as estrogen receptor modulators for the treatment of cancer.

Description

ESTROGEN RECEPTOR MODULATORS AND USES THEREOF
RELATED APPLICATIONS
[0001] This application claims priority to and benefit of U.S. Application No. 63/399,276, filed August 19, 2022, the entire contents of which are hereby incorporated by reference.
BACKGROUND
[0002] The estrogen receptor (ER) plays important roles in various diseases, disorders, and conditions, such as cancers, including breast cancers, menopause-related conditions or symptoms, and osteoporosis. An, K-C. Asian Spine J. 10(4);787-91 (2016 Aug). About 70% of human breast cancers are hormone dependent and ER-positive. Lumachi, et al., Curr. Med. Chem., 20(5):596-604 (2013). A variety of treatments have been developed to target the estrogen receptor and/or its activities.
SUMMARY
[0003] Selective estrogen receptor modulators (SERMs) and/or degraders (SERDs) are a particularly useful or promising tools for such therapy. For example, an estrogen receptor modulator that acts as an agonist (or partial agonist) in bone tissue may be useful for treating osteoporosis, e.g., in post-menopausal women. Further, an estrogen receptor modulator that acts as an antagonist in breast tissue may be useful for treating breast cancer. In some instances, the same estrogen receptor modulator may be used in both scenarios.
[0004] In some embodiments, the present disclosure provides compounds that are estrogen receptor modulators. In some embodiments, provided compounds are estrogen receptor agonists, e.g., as defined herein. In some embodiments, provided compounds are estrogen receptor antagonists, e.g., as defined herein.
[0005] Additionally, there remains a need for anti-estrogen agents that can completely inhibit estrogen receptors, including those coded for by both wild-type and mutant versions (e.g., those containing activating mutations) of the gene encoding Estrogen Receptor-alpha (ERa), Estrogen Receptor 1 (ESRI). The estrogen receptor is a tripartite protein comprising two distinct transcriptional activation functions (AF1 and AF2). Complete anti-estrogen activity requires inactivation of both AF1 and AF2. Activating mutations in the gene that codes for estrogen receptor 1 allows for activation of both AF1 and AF2 even in the absence of estrogen.
[0006] Many patients develop resistance to certain therapies that target the estrogen receptor (ER) over time. Certain first line therapies for treating ER-associated diseases, disorders, or conditions, are found to exhibit agonistic activity in conjunction with their antagonistic properties. Fulvestrant, in contrast, is the only approved therapy that exhibits complete antiestrogenic activity, but is not orally bioavailable, and must be administered parenterally.
[0007] In some embodiments, the present disclosure provides certain compounds and compositions that are complete estrogen receptor antagonists, and therefore do not suffer from the deficiencies found in previous therapies.
[0008] Additionally, in some embodiments, provided compounds may be orally bioavailable. [0009] In some embodiments, the present disclosure provides an estrogen receptor modulator (e.g., an estrogen receptor agonist, an estrogen receptor antagonist, and/or a complete estrogen receptor antagonist) that is a compound of Formula I:
Figure imgf000004_0001
or a pharmaceutically acceptable salt thereof, wherein A, B, L, n, R1, R2, R3, and R4 are as defined herein.
[0010] In some embodiments, the present disclosure provides methods of treating a disease, disorder, or condition associated with an estrogen receptor. In some embodiments, the present disclosure provides a methods of treating a disease, disorder, or condition associated with a mutation of an estrogen receptor.
[0011] In some embodiments, the present disclosure provides methods of treating a cancer. In some embodiments, the present disclosure provides methods of treating a cancer comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof, optionally in combination with an anti-cancer agent.
[0012] In some embodiments, the present disclosure provides methods of preventing recurrence of a cancer. In some embodiments, the present disclosure provides methods of preventing recurrence of a cancer comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof, optionally in combination with an anti-cancer agent.
[0013] In some embodiments, the present disclosure provides methods of treating osteoporosis, e.g., in post-menopausal women. In some embodiments, the present disclosure provides methods of treating osteoporosis comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof.
[0014] In some embodiments, the present disclosure provides methods of treating one or more menopausal symptoms or conditions. In some embodiments, the present disclosure provides methods of treating one or more menopausal symptoms comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS
[0015] The present disclosure provides compounds and compositions useful as estrogen receptor modulators (e.g., estrogen receptor agonists, estrogen receptor antagonists, and/or complete estrogen receptor antagonists). In some embodiments, such compounds include those of the formulae described herein, or a pharmaceutically acceptable salt thereof, wherein each variable is as defined and described herein.
Compounds and Definitions
[0016] Compounds of this disclosure include those described generally above and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75111 Ed. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.
[0017] Unless otherwise stated, structures depicted herein are meant to include all stereoisomeric (e.g., enantiomeric or diastereomeric) forms of the structure, as well as all geometric or conformational isomeric forms of the structure. For example, the R and S configurations of each stereocenter are contemplated as part of the disclosure. Therefore, single stereochemical isomers, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of provided compounds are within the scope of the disclosure. For example, in some cases, Table 1 shows one or more stereoisomers of a compound, and unless otherwise indicated, represents each stereoisomer alone and/or as a mixture. Unless otherwise stated, all tautomeric forms of provided compounds are within the scope of the disclosure.
[0018] Unless otherwise indicated, structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including replacement of hydrogen by deuterium or tritium, or replacement of a carbon by 13C- or 14C-enriched carbon are within the scope of this disclosure.
[0019] About or approximately: As used herein, the term "approximately" or "about," as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In general, those skilled in the art, familiar within the context, will appreciate the relevant degree of variance encompassed by "about" or "approximately" in that context. For example, in some embodiments, the term "approximately" or "about" may encompass a range of values that are within (i.e., ±) 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.
[0020] Administering: As used herein, the term "administering" or "administration" typically refers to the administration of a composition to a subject to achieve delivery of an agent that is, or is included in, a composition to a target site or a site to be treated. Those of ordinary skill in the art will be aware of a variety of routes that may, in appropriate circumstances, be utilized for administration to a subject, for example a human. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, etc. In some particular embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, dermal (which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreal, etc. In some embodiments, administration may be parenteral. In some embodiments, administration may be oral. In some particular embodiments, administration may be intravenous. In some particular embodiments, administration may be subcutaneous. In some embodiments, administration may involve only a single dose. In some embodiments, administration may involve application of a fixed number of doses. In some embodiments, administration may involve dosing that is intermittent (e.g., a plurality of doses separated in time) and/or periodic (e.g., individual doses separated by a common period of time) dosing. In some embodiments, administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time. In some embodiments, administration may comprise a prime-and-boost protocol. A prime-and-boost protocol can include administration of a first dose of a pharmaceutical composition followed by, after an interval of time, administration of a second or subsequent dose of a pharmaceutical composition.
[0021] Agonist: As used herein, the term “agonist” generally refers to an agent whose presence or level correlates with elevated level or activity of a target, as compared with that observed absent the agent (or with the agent at a different level). In some embodiments, an agonist is one whose presence or level correlates with a target level or activity that is comparable to or greater than a particular reference level or activity (e.g., that observed under appropriate reference conditions, such as presence of a known agonist, e.g., a positive control). In some embodiments, an agonist may be a direct agonist in that it exerts its influence directly on (e.g., interacts directly with) the target; in some embodiments, an agonist may be an indirect agonist in that it exerts its influence indirectly (e.g., by acting on, such as interacting with, a regulator of the target, or with some other component or entity.
[0022] Aliphatic: The term “aliphatic” refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “cycloaliphatic”), that has a single point or more than one points of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-12 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms (e.g., Ci-e). In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms (e.g., C1-5). In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms (e.g., C1-4). In still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms (e.g., C1-3), and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms (e.g., C1-2). Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups and hybrids thereof. A preferred aliphatic group is C1-6 alkyl.
[0023] Alkyl: The term “alkyl”, used alone or as part of a larger moiety, refers to a saturated, optionally substituted straight or branched chain hydrocarbon group having (unless otherwise specified) 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms (e.g., C1-12, C1-10, Ci-s, C1-6, Ci-4, Ci- 3, or C1-2). Exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, and heptyl.
[0024] Alkylene: The term “alkylene” is refers to a bivalent alkyl group. In some embodiments, “alkylene” is a bivalent straight or branched alkyl group. In some embodiments, an "alkylene chain" is a polymethylene group, i.e., -(CH2)n-, wherein n is a positive integer, e.g., from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. An optionally substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms is optionally replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group and also include those described in the specification herein. It will be appreciated that two substituents of the alkylene group may be taken together to form a ring system. In certain embodiments, two substituents can be taken together to form a 3- to 7- membered ring. The substituents can be on the same or different atoms. The suffix “-ene” when appended to certain groups herein are intended to refer to a bifunctional moiety of said group. For example, “-ene”, when appended to “cyclopropyl” becomes “cyclopropylene” and is intended to refer to a bifunctional cyclopropyl group, e.g.,
Figure imgf000008_0001
[0025] Alkenyl: The term “alkenyl”, used alone or as part of a larger moiety, refers to an optionally substituted straight or branched chain or cyclic hydrocarbon group having at least one double bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms(e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or C2-3). Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl. The term “cycloalkenyl” refers to an optionally substituted non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon double bond and having about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.
[0026] Alkynyl: The term “alkynyl”, used alone or as part of a larger moiety, refers to an optionally substituted straight or branched chain hydrocarbon group having at least one triple bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms (e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or C2-3). Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.
[0027] Antagonist. As will be understood by those skilled in the art, the term “antagonist” generally refers to an agent whose presence or level correlates with decreased level or activity of a target, as compared with that observed absent the agent (or with the agent at a different level). In some embodiments, an antagonist is one whose presence or level correlates with a target level or activity that is comparable to or less than a particular reference level or activity (e.g., that observed under appropriate reference conditions, such as presence of a known antagonist, e.g., a positive control). In some embodiments, an antagonist may be a direct antagonist in that it exerts its influence directly on (e.g., interacts directly with) the target; in some embodiments, an antagonist may be an indirect antagonist in that it exerts its influence indirectly (e.g., by acting on, such as interacting with, a regulator of the target, or with some other component or entity.
[0028] Aryl: The term “aryl” refers to monocyclic and bicyclic ring systems having a total of six to fourteen ring members (e.g., C6-C14), wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. In some embodiments, an “aryl” group contains between six and twelve total ring members (e.g., C6-C12). The term “aryl” may be used interchangeably with the term “aryl ring”. In certain embodiments, “aryl” refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Unless otherwise specified, “aryl” groups are hydrocarbons. In some embodiments, an “aryl” ring system is an aromatic ring (e.g., phenyl) that is fused to a non-aromatic ring (e.g., cycloalkyl). Examples of aryl rings include that are fused include
Figure imgf000009_0001
[0029] Biological sample: As used herein, the term “biological sample” typically refers to a sample obtained or derived from a biological source (e.g., a tissue or organism or cell culture) of interest, as described herein. In some embodiments, a source of interest comprises an organism, such as an animal or human. In some embodiments, a biological sample is or comprises biological tissue or fluid. In some embodiments, a biological sample may be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; cell-containing body fluids; free floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; washings or lavages such as a ductal lavages or broncheoalveolar lavages; aspirates; scrapings; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions, and/or excretions; and/or cells therefrom, etc. In some embodiments, a biological sample is or comprises cells obtained from an individual. In some embodiments, obtained cells are or include cells from an individual from whom the sample is obtained. In some embodiments, a sample is a “primary sample” obtained directly from a source of interest by any appropriate means. For example, in some embodiments, a primary biological sample is obtained by methods selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluid (e.g., blood, lymph, feces etc.), etc. In some embodiments, as will be clear from context, the term “sample” refers to a preparation that is obtained by processing (e.g., by removing one or more components of and/or by adding one or more agents to) a primary sample. For example, filtering using a semi-permeable membrane. Such a “processed sample” may comprise, for example, nucleic acids or proteins extracted from a sample or obtained by subjecting a primary sample to techniques such as amplification or reverse transcription of mRNA, isolation and/or purification of certain components, etc.
[0030] Carrier: As used herein, the term “carrier” refers to a diluent, adjuvant, excipient, or vehicle with which a composition is administered. In some exemplary embodiments, carriers can include sterile liquids, such as, for example, water and oils, including oils of petroleum, animal, vegetable or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil and the like. In some embodiments, carriers are or include one or more solid components.
[0031] Combination therapy: As used herein, the term “combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents or modality(ies)). In some embodiments, the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, “administration” of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination. For clarity, combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some embodiments, two or more agents, or active moieties thereof, may be administered together in a combination composition, or even in a combination compound (e.g., as part of a single chemical complex or covalent entity).
[0032] Comparable As used herein, the term “comparable” refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison therebetween so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features. Those of ordinary skill in the art will understand, in context, what degree of identity is required in any given circumstance for two or more such agents, entities, situations, sets of conditions, etc. to be considered comparable. For example, those of ordinary skill in the art will appreciate that sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied.
[0033] Composition: Those skilled in the art will appreciate that the term “composition” may be used to refer to a discrete physical entity that comprises one or more specified components. In general, unless otherwise specified, a composition may be of any form - e.g., gas, gel, liquid, solid, etc.
[0034] Cycloaliphatic. As used herein, the term “cycloaliphatic” refers to a monocyclic C3-8 hydrocarbon or a bicyclic C5-10 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point or more than one points of attachment to the rest of the molecule.
[0035] Cycloalkyl. As used herein, the term “cycloalkyl” refers to an optionally substituted saturated monocyclic or polycyclic ring system of about 3 to about 10 ring carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.
[0036] Dosage form or unit dosage form: Those skilled in the art will appreciate that the term “dosage form” may be used to refer to a physically discrete unit of an active agent (e.g., a therapeutic or diagnostic agent) for administration to a subject. Typically, each such unit contains a predetermined quantity of active agent. In some embodiments, such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen).
[0037] Dosing regimen or therapeutic regimen: Those skilled in the art will appreciate that the terms “dosing regimen” and “therapeutic regimen” may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen comprises a plurality of doses each of which is separated in time from other doses. In some embodiments, individual doses are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen). [0038] Excipient: As used herein, the term “excipient” refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.
[0039] Heteroaliphatic. The term “heteroaliphatic” or “heteroaliphatic group”, as used herein, denotes an optionally substituted hydrocarbon moiety having, in addition to carbon atoms, from one to five heteroatoms, that may be straight-chain (i.e., unbranched), branched, or cyclic (“heterocyclic”) and may be completely saturated or may contain one or more units of unsaturation, but which is not aromatic. The term “heteroatom” refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quatemized form of a basic nitrogen. The term “nitrogen” also includes a substituted nitrogen. Unless otherwise specified, heteroaliphatic groups contain 1-10 carbon atoms wherein 1-3 carbon atoms are optionally and independently replaced with heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, heteroaliphatic groups contain 1-4 carbon atoms, wherein 1-2 carbon atoms are optionally and independently replaced with heteroatoms selected from oxygen, nitrogen, and sulfur. In yet other embodiments, heteroaliphatic groups contain 1-3 carbon atoms, wherein 1 carbon atom is optionally and independently replaced with a heteroatom selected from oxygen, nitrogen, and sulfur. Suitable heteroaliphatic groups include, but are not limited to, linear or branched, heteroalkyl, heteroalkenyl, and heteroalkynyl groups. For example, a 1- to 10 atom heteroaliphatic group includes the following exemplary groups: -O-CH3, -CH2-O-CH3, -O-CH2- CH2-O-CH2-CH2-O-CH3, and the like.
[0040] Heteroaryl: The terms “heteroaryl” and “heteroar-”, used alone or as part of a larger moiety, e.g., “heteroaralkyl”, or “heteroaralkoxy”, refer to monocyclic or bicyclic ring groups having 5 to 10 ring atoms (e.g., 5- to 6-membered monocyclic heteroaryl or 9- to 10-membered bicyclic heteroaryl); having 6, 10, or 14 71-electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[l,2-a]pyrimidinyl, imidazo[l,2-a]pyridyl, imidazo[4,5-b]pyridyl, imidazo[4,5-c]pyridyl, pyrrol opyridyl, pyrrolopyrazinyl, thienopyrimidinyl, triazolopyridyl, and benzoisoxazolyl. The terms “heteroaryl” and “heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms). Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzotri azolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4/7- quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3-b]-l,4-oxazin-3(4H)-one, 4H- thieno[3,2-b]pyrrole, and benzoisoxazolyl. The term “heteroaryl” may be used interchangeably with the terms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any of which terms include rings that are optionally substituted.
[0041] Heteroatom: The term “heteroatom” as used herein refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quatemized form of a basic nitrogen.
[0042] Heterocycle: As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a stable 3- to 8-membered monocyclic, a 6- to 10-membered bicyclic, or a 10- to 16-membered polycyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, such as one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR+ (as in N-substituted pyrrolidinyl). A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiamorpholinyl. A heterocyclyl group may be mono-, bi-, tri-, or polycyclic, preferably mono-, bi-, or tricyclic, more preferably mono- or bicyclic. A bicyclic heterocyclic ring also includes groups in which the heterocyclic ring is fused to one or more aryl rings. Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl. A bicyclic heterocyclic ring can also be a spirocyclic ring system (e.g., 6- to 11 -membered spirocyclic fused heterocyclic ring having, in addition to carbon atoms, one or more heteroatoms as defined above (e.g., one, two, three or four heteroatoms)). A bicyclic heterocyclic ring can also be a bridged ring system (e.g., 7- to 11 -membered bridged heterocyclic ring having one, two, or three bridging atoms.
[0043] Modulator. The term “modulator,” as used herein, refers to a compound (e.g., a small molecule) that can alter the activity of another molecule (e.g., a protein). For example, in some embodiments, a modulator can cause an increase or decrease in the magnitude of a certain activity of a type of molecule as compared to the magnitude of the activity in the absence of the modulator. For example, a modulator can be an agonist or an antagonist of a particular target, as those terms are defined herein. For example, in some embodiments, a modulator is an agonist. In some embodiments, a modulator is an antagonist.
[0044] Oral: The phrases “oral administration” and “administered orally” as used herein have their art-understood meaning referring to administration by mouth of a compound or composition.
[0045] Parenteral: The phrases “parenteral administration” and “administered parenterally” as used herein have their art-understood meaning referring to modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal, and intrastemal injection and infusion.
[0046] Partially unsaturated: As used herein, the term “partially unsaturated” refers to a ring moiety that includes at least one double or triple bond between ring atoms. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (e.g., aryl or heteroaryl) moieties, as herein defined.
[0047] Patient or subject: As used herein, the term “patient” or “subject” refers to any organism to which a provided composition is or may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients or subjects include animals (e g., mammals such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, a patient is a human. In some embodiments, a patient or a subject is suffering from or susceptible to one or more disorders or conditions. In some embodiments, a patient or subject displays one or more symptoms of a disorder or condition. In some embodiments, a patient or subject has been diagnosed with one or more disorders or conditions. In some embodiments, a patient or a subject is receiving or has received certain therapy to diagnose and/or to treat a disease, disorder, or condition.
[0048] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in unit dose amount appropriate for administration in a therapeutic or dosing regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonary, and to other mucosal surfaces.
[0049] Pharmaceutically acceptable: As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
[0050] Pharmaceutically acceptable salt: The term “pharmaceutically acceptable salt”, as used herein, refers to salts of such compounds that are appropriate for use in pharmaceutical contexts, i.e., salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977).
[0051] Prevent or prevention: As used herein, the terms “prevent” or “prevention”, when used in connection with the occurrence of a disease, disorder, and/or condition, refer to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time.
[0052] Substituted or optionally substituted: As described herein, compounds of the invention may contain “optionally substituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. “Substituted” applies to one or more hydrogens that are either explicit or implicit from the structure (e.g., O RI refers to at least
Figure imgf000017_0001
Figure imgf000017_0002
). Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this invention are preferably those that result in the formation of stable or chemically feasible compounds. The term “stable,” as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes provided herein. Groups described as being “substituted” preferably have between 1 and 4 substituents, more preferably 1 or 2 substituents. Groups described as being “optionally substituted” may be unsubstituted or be “substituted” as described above.
[0053] Suitable monovalent substituents on a substitutable carbon atom of an “optionally substituted” group are independently halogen; -(CH2)o-4R°; -(CH2)OMOR°; -0(CH2)O-4R°, -O- (CH2)O-4C(0)OR°; -(CH2)O-4CH(OR°)2; -(CH2)O-4SRC; -('CFhjo-iPh, which may be substituted with R°; -(CH2)o IO(CH2)O iPh which may be substituted with R°; -CH=CHPh, which may be substituted with R°; -(CH2)o^O(CH2)o-i-pyridyl which may be substituted with R°; -NO2; -CN; -N3; -(CH2)O-4N(R°)2; -(CH2)O^N(R°)C(0)R°; -N(R°)C(S)R°; -(CH2)O-
4N(R°)C(O)NR°2; -N(R°)C(S)NR°2; -(CH2)O-4N(R°)C(0)OR°;
N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; -(CH2)o-4C(0)R°; - C(S)R°; -(CH2)O-4C(0)OR°; -(CH2)O-4C(0)SR°, -(CH2)o^C(0)OSiR°3; -(CH2)o^OC(0)R°; - OC(0)(CH2)O-4SR°; -(CH2)O-4SC(0)R°; -(CH2)O-4C(0)NR°2; -C(S)NR°2; -C(S)SR°; - SC(S)SR°, -(CH2)O-4OC(0)NR°2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)R°; - C(NOR°)R°; -(CH2)O-4SSR°; -(CH2)O-4S(0)2R°; -(CH2)O-4S(0)20R°; -(CH2)O^OS(0)2R°; - S(O)2NR°2; -(CH2)O^S(0)R°; -N(R°)S(O)2NR°2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NH)NR°2; - P(O)2R°; -P(O)R°2; -OP(O)R°2; -OP(O)(OR°)2; -SiR°3; -(Ci-4 straight or branched alkylene)O- N(R°)2; or — (C1-4 straight or branched alkylene)C(O)O-N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, C1-6 aliphatic, -CH2Ph, -0(CH2)o- iPh, -CH2-(5- to 6-membered heteroaryl ring), or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3- to 12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.
[0054] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), are independently halogen, -(CH2)o-2R*, -(haloR*), -(CH2)o-2OH, -(CH2)o-2OR*, -(CH2)o-
2CH(OR’)2, -O(haloR’), -CN, -N3, -(CH2)o-2C(0)R*, -(CH2)o-2C(0)OH, -(CH2)o-2C(0)OR*, - (CH2)O-2SR*, -(CH2)O-2SH, -(CH2)O-2NH2, -(CH2)o-2NHRe, -(CH2)O-2NR’2, -NO2, -SiR*3, - OSiR*3, -C(O)SR’, -(Ci~ i straight or branched alkylene)C(O)OR*, or -SSR* wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently selected from C1-4 aliphatic, -CJLPh, -0(CH2)o-iPh, or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S.
[0055] Suitable divalent substituents on a saturated carbon atom of an “optionally substituted” group include the following: =0 (“oxo”), =S, =NNR*2, =NNHC(O)R*, =NNHC(0)0R*, =NNHS(O)2R*, =NR*, =N0R*, -O(C(R*2))2-3O-, or -S(C(R*2))2-3S-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: -O(CR*2)2-3O-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0056] Suitable substituents on the aliphatic group of R* include halogen, - R*, -(haloR*), -OH, -OR*, -O(haloR’), -CN, -C(O)OH, -C(O)OR’, -NH2, -NHR*, -NR*2, or -NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1-4 aliphatic, -CH2Ph, -0(CH2)o iPh, or a 3- to 6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0057] Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include -Rf, -NRf 2, -C(O)Rf, -C(O)ORf, -C(O)C(O)Rf, C(O)CH2C(O)Rt, -S(O)2RT, -SCO^NR^, -C(S)NRf 2, -CCNT^NR^, or -NCR^SCO)^; wherein each R' is independently hydrogen, C1-6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R', taken together with their intervening atom(s) form an unsubstituted 3- to 12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0058] Suitable substituents on the aliphatic group of R? are independently halogen, - R*, -(haloR*), -OH, -OR’, -O(haloR’), -CN, -C(O)OH, -C(O)OR*, -NH2, -NHR*, -NR*2, or -NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C 1-4 aliphatic, -CH2Ph, -0(CH2)o-iPh, or a 3- to 6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
[0059] Small molecule: As used herein, the term “small molecule” means a low molecular weight organic and/or inorganic compound. In general, a “small molecule” is a molecule that is less than about 5 kilodaltons (kD) in size. In some embodiments, a small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, the small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, a small molecule is less than about 2000 g/mol, less than about 1500 g/mol, less than about 1000 g/mol, less than about 800 g/mol, or less than about 500 g/mol. In some embodiments, a small molecule is not a polymer.
[0060] In some embodiments, a small molecule does not include a polymeric moiety. In some embodiments, a small molecule is not and/or does not comprise a protein or polypeptide (e.g., is not an oligopeptide or peptide). In some embodiments, a small molecule is not and/or does not comprise a polynucleotide (e.g., is not an oligonucleotide). In some embodiments, a small molecule is not and/or does not comprise a polysaccharide; for example, in some embodiments, a small molecule is not a glycoprotein, proteoglycan, glycolipid, etc.). In some embodiments, a small molecule is not a lipid.
[0061] In some embodiments, a small molecule is a modulating agent (e.g., is an inhibiting agent or an activating agent). In some embodiments, a small molecule is biologically active. In some embodiments, a small molecule is detectable (e.g., comprises at least one detectable moiety). In some embodiments, a small molecule is a therapeutic agent.
[0062] Those of ordinary skill in the art, reading the present disclosure, will appreciate that certain small molecule compounds described herein may be provided and/or utilized in any of a variety of forms such as, for example, crystal forms (e.g., polymorphs, solvates, etc), salt forms, protected forms, pro-drug forms, ester forms, isomeric forms (e.g., optical and/or structural isomers), isotopic forms, etc.
[0063] Those of ordinary skill in the art will appreciate that certain small molecule compounds have structures that can exist in one or more steroi someric forms. In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in the form of an individual enantiomer, diastereomer or geometric isomer, or may be in the form of a mixture of stereoisomers; in some embodiments, such a small molecule may be utilized in accordance with the present disclosure in a racemic mixture form.
[0064] Those of skill in the art will appreciate that certain small molecule compounds have structures that can exist in one or more tautomeric forms. In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in the form of an individual tautomer, or in a form that interconverts between tautomeric forms.
[0065] Those of skill in the art will appreciate that certain small molecule compounds have structures that permit isotopic substitution (e.g., 2H or 3H for H; nC, 13C or 14C for 12C; 13N or 15N for 14N; 17O or 18O for 16O; 36C1 for 35C1 or j7Cl; 18F for 19F; 131I for 127I; etc.). In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in one or more isotopically modified forms, or mixtures thereof.
[0066] In some embodiments, reference to a particular small molecule compound may relate to a specific form of that compound. In some embodiments, a particular small molecule compound may be provided and/or utilized in a salt form (e.g., in an acid-addition or baseaddition salt form, depending on the compound); in some such embodiments, the salt form may be a pharmaceutically acceptable salt form.
[0067] In some embodiments, where a small molecule compound is one that exists or is found in nature, that compound may be provided and/or utilized in accordance in the present disclosure in a form different from that in which it exists or is found in nature. Those of ordinary skill in the art will appreciate that, in some embodiments, a preparation of a particular small molecule compound that contains an absolute or relative amount of the compound, or of a particular form thereof, that is different from the absolute or relative (with respect to another component of the preparation including, for example, another form of the compound) amount of the compound or form that is present in a reference preparation of interest (e.g., in a primary sample from a source of interest such as a biological or environmental source) is distinct from the compound as it exists in the reference preparation or source. Thus, in some embodiments, for example, a preparation of a single stereoisomer of a small molecule compound may be considered to be a different form of the compound than a racemic mixture of the compound; a particular salt of a small molecule compound may be considered to be a different form from another salt form of the compound; a preparation that contains only a form of the compound that contains one conformational isomer ((Z) or (E)) of a double bond may be considered to be a different form of the compound from one that contains the other conformational isomer ((E) or (Z)) of the double bond; a preparation in which one or more atoms is a different isotope than is present in a reference preparation may be considered to be a different form; etc.
[0068] Those skilled in the art will further appreciate that, in small molecule structures, the symbol as used herein, refers to a point of attachment between two atoms. Additionally or alternatively, the symbol
Figure imgf000022_0001
refers to a point of attachment ring in a spirocyclic manner
[0069] Treat: As used herein, the terms “treat,” “treatment,” or “treating” refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition, for example, for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition.
Estrogen Receptor Modulators
[0070] In some embodiments, the present disclosure provides compounds that are estrogen receptor modulators. In some embodiments, provided compounds are estrogen receptor agonists. As used herein, an “estrogen receptor agonist” refers to a compound or composition that produces an agonistic effect when contacting the estrogen receptor of a subject or biological sample. In some embodiments, an estrogen receptor agonist is characterized by having (i) at least 80% increase in the E2 -normalized signal in the AP assay (agonist mode) of Example 209 and (ii) no more than 80% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209. In some embodiments, provided compounds are estrogen receptor antagonists. As used herein, an “estrogen receptor antagonist” refers to a compound or composition that produces an antagonistic effect when contacting the estrogen receptor of a subject or biological sample. In some embodiments, an estrogen receptor antagonist is characterized by having:
1. (i) between 10% and 80% increase in the E2-normalized signal in the AP assay (agonist mode) of Example 209 and (ii) between 10% and 80% reduction in the E2- normalized signal in the AP assay (antagonist mode) of Example 209;
2. (i) a pIC50 greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode) of Example 209; or
3. (i) a pICso greater than 7.5 and at least a 10% reduction in the E2 -normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode) of Example 209.
[0071] In some embodiments, the present disclosure provides compounds that are complete estrogen receptor (ER) antagonists. As used herein, a “complete estrogen receptor antagonist” refers to a compound or composition that produces an antagonistic effect when contacting the estrogen receptor of a subject or biological sample, with minimal agonistic effect (e g., with no or substantially no agonistic effect). Complete estrogen receptor antagonism is determined according to methods described herein, for example in Example 209. In some embodiments, a complete estrogen receptor antagonist is characterized by having (i) a pICso greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode) of Example 209. In some embodiments, a complete estrogen receptor antagonist is characterized by having (i) a pICso greater than 7.5 and at least a 10% reduction in the E2- normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode) of Example 209. In some embodiments, a complete estrogen receptor antagonist is an agent (e.g., a small molecule compound) that shows ER antagonism and no or substantially no ER agonism in one or more of ERct protein level assays, MCF-7 cell line assays, Ishikawa cell line assays (measuring wild type ER and certain mutants including mutants lacking AF1 and/or AF2 domains), and rodent uterine weight gain assays. See, generally, WO 2017/059139. Alternatively or additionally, in some embodiments, a complete estrogen receptor antagonist has three characteristics: it (1) inhibits both activating function 1 (AF1) and activating function 2 (AF2), as complete anti-estrogen activity requires inactivation of both AF1 and AF2; (2) promotes ER degradation; and (3) lacks the partial ER agonist activity observed with certain other agents. Without being bound by theory, it is understood that complete inhibition of both AF1 and AF2 is required for complete estrogen receptor activity, activating mutations in the gene that codes for estrogen receptor 1 allows for activation of both AF1 and AF2 even in the absence of estrogen. [0072] In some embodiments, the present disclosure provides a compound of Formula I:
Figure imgf000024_0001
or a pharmaceutically acceptable salt thereof, wherein: A is an optionally substituted 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S or an optionally substituted 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S; L is a covalent bond or an optionally substituted bivalent group selected from -C1-C6 aliphatic-, - La-C0-C5 aliphatic-, and -C1-C5 aliphatic-La-, wherein La is selected from -S-, -SO-, -SO2-, and -N(Ra)-; B is selected from –OH, -CO2H, C1-C6 aliphatic, 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S, and C3-C6 cycloaliphatic; R1 is selected from hydrogen and optionally substituted C1-C6 aliphatic; R2 is selected from hydrogen and optionally substituted C1-C6 aliphatic; R3 is selected from hydrogen, halogen, -CN, -ORa, -C(O)Ra, -C(O)2Ra, -OC(O)Ra , -C(O)N(Ra)2, -OC(O)N(Ra)2, -NO2, -N(Ra)2, -N(Ra)C(O)Ra, -N(Ra)C(O)2Ra, -N(Ra)S(O)2Ra, -SRa, - S(O)2Ra, -S(O)N(Ra)2, -S(O)2N(Ra)2, and an optionally substituted C1-6 aliphatic group; each R4 is independently oxo, halogen, -CN, -ORa, -N(Ra)2, -C(O)Ra, -OC(O)Ra, -C(O)2Ra, - C(O)N(Ra)2, -N(Ra)C(O)Ra, or an optionally substituted group selected from C1-C6 aliphatic and 3- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S; each Ra is independently selected from hydrogen and optionally substituted Ci-Ce aliphatic; and n is 0 to 5.
[0073] As defined generally above, A is an optionally substituted 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S or an optionally substituted 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, A is 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S or 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S, each optionally substituted with halogen, -(CH2)o^R°, or -(CH2)O-40R°.
[0074] In some embodiments, A is optionally substituted 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S optionally substituted with halogen, -(CH2)o-4R°, or -(CH2)o-40R°. In some embodiments, A is 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S, optionally substituted with halogen or Ci-Ce aliphatic. In some embodiments, A is optionally substituted 7-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 8-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 9- membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S.
[0075] In some embodiments, A is
Figure imgf000026_0001
attachment to moiety L.
[0076] In some embodiments, A is
Figure imgf000027_0001
[0078] In some embodiments, A is
Figure imgf000028_0001
[0079] In some embodiments, A is optionally substituted 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, A is optionally substituted 4- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, A is 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S optionally substituted with halogen, -(CH2)o-4R°, or -(CI l2 )o-40R°. In some embodiments, A is 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S comprising 1-4 heteroatoms selected from N, O, and S, optionally substituted with halogen or Ci-Ce aliphatic.
[0080] In some embodiments, A is optionally substituted piperidinyl. In some embodiments, A is:
Figure imgf000028_0002
, wherein * represents a point of attachment to moiety L.
[0081] As described generally above, L is a covalent bond or an optionally substituted bivalent group selected from Ci-Ce aliphatic, -La-Co-C5 aliphatic-, and -C1-C5 aliphatic-!?-, wherein La is selected from -S-, -SO-, -SO2-, and -N(Ra)-. It will be appreciated that the point of attachment of L is indicated by the direction in which its definition is written, such that the leftmost atom is attached to A and the rightmost atom is attached to B. For example, when L is - S-CH2-, L is attached to A via the sulfur atom and to B via the carbon atom.
[0082] In some embodiments, L is a covalent bond.
[0083] In some embodiments, L is optionally substituted Ci-Ce aliphatic. In some embodiments, L is -(CH2)I-6-. In some embodiments, L is -CH2-, -CH2-CH2-, -CH2-CH2-, -CH2- CH2-CH2-, -CH2-CH2-CH2-CH2-, -CH2-CH2-CH2-CH2-CH2-, or -CH2-CH2-CH2-CH2-CH2-CH2-. In some embodiments, L is -CH2-.
[0084] In some embodiments, L is optionally substituted -La-Co-Cs aliphatic. In some embodiments, L is -S-C0-C5 aliphatic, -SO-C0-C5 aliphatic, -SO2-C0-C5 aliphatic, or -N(Ra)-Co- C5 aliphatic. In some embodiments, L is -S-C0-C5 aliphatic. In some embodiments, L is -SO-Co- Cs aliphatic. In some embodiments, L is -SO2-C0-C5 aliphatic. In some embodiments, L is - N(Ra)-Co-Cs aliphatic. In some embodiments, L is La. In some embodiments, L is -S-, -SO-, - SO2-, or -N(Ra)-. In some embodiments, L is -S-. In some embodiments, L is -SO-. In some embodiments, L is -SO2-. In some embodiments, L is -N(Ra)-. In some embodiments, L is - N(H)-. In some embodiments, L is -N(CH3)-.
[0085] In some embodiments, L is selected from a covalent bond, -CH2-, -CH2-CH2-, -CH2- CH2-CH2-, -S-, and -SO2-. In some embodiments, L is selected from a covalent bond, -CH2-, - CH2-CH2-, -CH2-CH2-CH2-, -S-, and -N(H)-.
[0086] As described generally above, B is selected from -OH, -CO2H, Ci-Ce aliphatic, 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S, and C3-C6 cycloaliphatic. In some embodiments, when B is -OH or -CO2H, n is 0.
[0087] In some embodiments, B is selected from 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S and C3-C6 cycloaliphatic. In some embodiments, B is selected from -OH, -CO2H, Ci-Ce aliphatic, and 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is selected from -CO2H and 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S.
[0088] In some embodiments, B is -OH.
[0089] In some embodiments, B is -CO2H.
[0090] In some embodiments, B is Ci-Ce aliphatic. In some embodiments, B is Ci-Ce alkyl.
[0091] In some embodiments, B is 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is a 4- to 7-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is a 4- or 7-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S.
[0092] In some embodiments, B is 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is 3-membered heterocyclyl comprising 1 heteroatom selected from N, O, and S. In some embodiments, B is 4-membered heterocyclyl comprising 1-2 heteroatoms selected from N, O, and S. In some embodiments, B is 5-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. In some embodiments, B is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or Attorney Docket No.: 2012034-0263 morpholinyl. In some embodiments, B is azetidinyl or pyrrolidinyl. In some embodiments, B is azetidinyl. In some embodiments, B is pyrrolidinyl. In some embodiments, B is selected from: ,
Figure imgf000030_0001
[0093] In some embodiments, B is selected from: and
Figure imgf000030_0002
.
Figure imgf000030_0003
[0094] In some embodiments, B is selected from In
Figure imgf000030_0004
some embodiments, B is . [0095] In some em
Figure imgf000030_0005
is 6- to 12-membered bicyclic fused or spirocyclic heterocyclyl. In some embodiments, B is 6- to 8-membered bicyclic fused or spirocyclic heterocyclyl. In some embodiments, B is 6- to 12-membered bicyclic fused heterocyclyl. In some embodiments, B is 6- to 8-membered bicyclic fused heterocyclyl. In some embodiments, B is 6- to 12-membered bicyclic spirocyclic heterocyclyl. In some embodiments, B is 6- to 8- membered bicyclic spirocyclic heterocyclyl. In some embodiments, B is selected from: , and
Figure imgf000030_0006
Page 28 of 231 11528310v1 [0096] In some embodiments, B is selected from:
Figure imgf000031_0001
Figure imgf000031_0002
[0097] In some embodiments, B is selected from:
Figure imgf000031_0003
In some embodiments, B is:
Figure imgf000031_0004
[0098] In some embodiments, B is Ca-Ce cycloaliphatic In some embodiments, B is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
[0099] As described generally above, each R4 is independently oxo, halogen, -CN, -ORa, - N(Ra)2, -C(O)Ra, -OC(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)Ra, or an optionally substituted group selected from Ci-Ce aliphatic and 3- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S.
[0100] In some embodiments, R4 is oxo.
[0101] In some embodiments, R4 is halogen. In some embodiments, R4 is flouro.
[0102] In some embodiments, R4 is -CN.
[0103] In some embodiments, R4 is -ORa. In some embodiments, R4 is -OH. In some embodiments, R4 is -O-Ci-Ce aliphatic (e.g., -OCH3).
[0104] In some embodiments, R4 is -N(Ra)2. In some embodiments, R4 is -N(H)(Ra). In some embodiments, R4 is -NH2. In some embodiments, R4 is -N(H)Ci-Ce aliphatic. In some embodiments, R4 is -N(Ci-Ce aliphatic)2. In some embodiments, R4 is -N(H)CH3, - N(H)CH2CH3, -N(CH2CH3)2J -N(H)CH2CH2CH3, -N(CH3)2, -N(CH3)CH2CH3, or - N(CH3)CH2CH2CH3.
[0105] In some embodiments, R4 is -C(O)Ra. In some embodiments, R4 is -C(O)-Ci-C6 aliphatic optionally substituted with -(CH2)O^IOR°. In some embodiments, R4 is -C(O)CH3, - C(O)CH2OCH3, or -C(O)CH2CH2OCH3.
[0106] In some embodiments, R4 is -OC(O)Ra. In some embodiments, R4 is -OC(O)-Ci-Ce aliphatic. [0107] In some embodiments, R4 is -C(O)2Ra In some embodiments, R4 is -C(O)OH. In some embodiments, R4 is -C(O)2-Ci-Ce aliphatic.
[0108] In some embodiments, R4 is -C(O)N(Ra)2. In some embodiments, R4 is - C(O)N(H)Ra. In some embodiments, R4 is -C(O)NH2. In some embodiments, R4 is - C(O)N(H)Ci-Ce aliphatic.
[0109] In some embodiments, R4 is -N(Ra)C(O)Ra. In some embodiments, R4 is - N(H)C(O)Ra. In some embodiments, R4 is -N(H)C(O)Ci-Ce aliphatic optionally substituted with -(CH2)O-40R°. In some embodiments, R4 is -N(H)C(O)CH2OH or -N(H)C(O)CH2OCH3.
[0110] In some embodiments, R4 is an optionally substituted Ci-Ce aliphatic. In some embodiments, R4 is Ci-Ce aliphatic optionally substituted with halogen, -(CH2)o-4R°, -(CH2)o- 4OR°, -0(CH2)o-4R°, -CN, -(CH2)O-4N(R°)2, or phenyl. In some embodiments, R4 is Ci-Ce aliphatic substituted with halogen, -OH, -OCH3, -CN, or 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R4 is Ci-Ce aliphatic optionally substituted with halogen. In some embodiments, R4 is C1-C3 alkyl optionally substituted with halogen. In some embodiments, R4 is methyl, ethyl, propyl, butyl, pentyl, hexyl, -CH2F, -CHF2, -CH2CHF2, - CH2CH2CHF2, -CH2CH2CF3, -CH2OCH3 -CH2CH2CH2OH, -CH2CH2CH2F, -CH2CH2CH(CH3)2, -CH(CH3)2, -CH2CH2Ph, -C(CH3)2-OH, -CH2CH2OCH3, -CH2CH2OCH2CH3, - CH2CH2CH2OCH2CH3, -CH2C=CH, -CH2C=CHCH3, -CH2CN, -CH2CH2CN,
Figure imgf000032_0001
in some embodiments, R4 is methyl, ethyl, propyl, -CH2F, -CHF2, -CH2OCH3, -C(CH3)2-OH, -CH(CH3)2, and -CH2CH2CH2F. In some embodiments, R4 is methyl, ethyl, propyl, -CH2F, -CH2CN, and - CH2CH2CH2F. In some embodiments, R4 is methyl or -CH2F.
[0111] In some embodiments, R4 is an optionally substituted 3- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S. In some embodiments, R4 is optionally substituted 3- to 6-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S. In some embodiments, R4 is optionally substituted azetidinyl.
[0112] In some embodiments, R4 is oxo, -OH, -OCH3, fluoro, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, -CN, -CH2F, -CHF2, -CH2CHF2, -CH2CH2CHF2, -CH2CH2CF3, - CH2CH2CH2OH, -CH2CH2CH2F, -CH2CH2CH(CH3)2, -CH2CH2-Ph, -C(CH3)2-OH, -CH2OCH3, 2OCH3, -CH2CH2OCH2CH3, -CH2CH2CH2OCH2CH3, -CH2C≡CH, -CH2C≡CHCH3, - CH2CN, -CH2CH2CN, -CH2CH2CH2OCF3, -NHCH2CH3, -NHCH2CH2CH3, -NHCH2CH2CH3, - NHCH3, -N(CH3)CH2CH3, -N(CH3)CH2CH2CH3, -N(CH3)2, -NHC(O)CH2OCH3, - , , -
Figure imgf000033_0001
OCH3, methyl, ethyl, propyl, -CH2F, -CH2OCH3, -C(CH3)2OH, -CH(CH3)2, and -CH2CH2CH2F. In some embodiments, each R4 is independently selected from fluoro, -N(CH3)2, methyl, ethyl, propyl, -CH2F, -CHF2, -CH2CN, and -CH2CH2CH2F. In some embodiments, each R4 is independently selected from fluoro, methyl, and -CH2F. [0114] In some embodiments, a moiety:
Figure imgf000033_0002
[0115] In some embodiments, a moiety:
Figure imgf000034_0001
is a moiety selected from:
Figure imgf000034_0002
[0116] In some embodiments, a moiety:
Figure imgf000034_0003
is a moiety selected from:
Figure imgf000034_0004
[0117] In some embodiments, a moiety:
Figure imgf000034_0005
is a moiety selected from:
Figure imgf000035_0001
[0118] In some embodiments, a moiety:
Figure imgf000035_0002
is a moiety selected from:
Figure imgf000035_0003
[0119] In some embodiments, a moiety:
Figure imgf000036_0001
Figure imgf000037_0001
Figure imgf000038_0001
[0120] In some embodiments, a moiety:
Figure imgf000038_0002
[0121] is a moiety selected from:
Figure imgf000038_0003
Figure imgf000038_0004
[0122] In some embodiments, a moiety:
Figure imgf000039_0001
is a moiety selected from:
Figure imgf000039_0002
Figure imgf000039_0003
[0123] In some embodiments, a moiety:
Figure imgf000039_0004
[0124] In some embodiments, a moiety:
Figure imgf000040_0001
is a moiety selected from:
Figure imgf000040_0002
[0125] As described generally above, R1 is selected from hydrogen and optionally substituted Ci-Ce aliphatic. In some embodiments, R1 is hydrogen. In some embodiments, R1 is Ci-Ce aliphatic optionally substituted with halogen, -(CH2)o-4R°, -(CH2)o^OR°, or -(CH2)o^iPh. In some embodiments, R1 is Ci-Ce aliphatic optionally substituted with halogen or -OH.
[0126] In some embodiments, R1 is selected from:
Figure imgf000040_0003
[0127] In some embodiments, R1 is selected from:
Figure imgf000040_0004
[0128] In some embodiments, R1 is selected from:
Figure imgf000040_0005
,
[0129] In some embodiments,
Figure imgf000040_0006
[0130] As described generally above, R2 is selected from hydrogen and optionally substituted
Ci-Ce aliphatic. In some embodiments, R2 is hydrogen. In some embodiments, R2 is Ci-Ce aliphatic. In some embodiments, R2 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments, R2 is methyl.
[0131] As described generally above, R3 is selected from hydrogen, halogen, -CN, -ORa, - C(O)Ra, -C(O)2Ra, -OC(O)Ra -C(O)N(Ra)2, -OC(O)N(Ra)2, -NO2, -N(Ra)2, -N(Ra)C(O)Ra, - N(Ra)C(O)2Ra, -N(Ra)S(O)2Ra, -SRa, -S(O)2Ra, -S(O)N(Ra)2, -S(O)2N(Ra)2, and an optionally substituted Ci-6 aliphatic group. In some embodiments, R3 is hydrogen. [0132] As described generally above, n is 0-5. In some embodiments, n is 0. In some embodiments, n is 1, 2, 3, 4, or 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.
[0133] In some embodiments, a compound of Formula I is a compound of Formula II:
Figure imgf000041_0001
or a pharmaceutically acceptable salt thereof, wherein A, B, L, n, R1, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination. [0134] In some embodiments, a compound of Formula I is a compound of Formula Il-a:
Figure imgf000041_0002
Il-a or a pharmaceutically acceptable salt thereof, wherein A, L, R1, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0135] In some embodiments, a compound of Formula I is a compound of Formula Il-b:
Figure imgf000042_0001
Il-b or a pharmaceutically acceptable salt thereof, wherein A, L, R1, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0136] In some embodiments, a compound of Formula I is a compound of Formula II-c:
Figure imgf000042_0002
or a pharmaceutically acceptable salt thereof, wherein A, L, R1, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0137] In some embodiments, a compound of Formula I is a compound of Formula Il-d:
Figure imgf000043_0001
Il-d or a pharmaceutically acceptable salt thereof, wherein A, L, R1, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0138] In some embodiments, a compound of Formula I is a compound of Formula Il-e:
Figure imgf000043_0002
Il-e or a pharmaceutically acceptable salt thereof, wherein A, L, R1, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0139] In some embodiments, a compound of Formula I is a compound of Formula Il-f:
Figure imgf000043_0003
or a pharmaceutically acceptable salt thereof, wherein A, B, L, n, R1, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination. [0140] In some embodiments, a compound of Formula I is a compound of Formula III:
Figure imgf000044_0001
or a pharmaceutically acceptable salt thereof, wherein B, L, n, R1, R2, R3, and R4 are as defined above for Formula T and described in classes and subclasses herein, both singly and in combination.
[0141] In some embodiments, a compound of Formula I is a compound of Formula IV:
Figure imgf000044_0002
or a pharmaceutically acceptable salt thereof, wherein B, L, n, R1, R2, R3, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0142] In some embodiments, a compound of Formula I is a compound of Formula V:
Figure imgf000045_0001
or a pharmaceutically acceptable salt thereof, wherein B, L, n, R1, R2, R3, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0143] In some embodiments, a compound of Formula I is a compound of Formula VI:
Figure imgf000045_0002
or a pharmaceutically acceptable salt thereof, wherein B, L, n, R1, R2, R3, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
[0144] In some embodiments, a compound of Formula I is a compound of Formula VII-1 or VII-2:
Figure imgf000045_0003
Figure imgf000046_0001
or a pharmaceutically acceptable salt thereof, wherein B, L, n, R1, R2, R3, and R4 are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination
[0145] In some embodiments, the present disclosure provides compounds selected from Table 1, or a pharmaceutically acceptable salt thereof:
Table 1
Figure imgf000046_0002
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Figure imgf000055_0001
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
Figure imgf000061_0001
Figure imgf000062_0001
Figure imgf000063_0001
Figure imgf000064_0001
Figure imgf000065_0001
Figure imgf000066_0001
Figure imgf000067_0002
[0146] In some embodiments, the present disclosure provides a compound of Table 2, or a pharmaceutically acceptable salt thereof:
Table 2
Figure imgf000067_0001
Figure imgf000068_0001
Figure imgf000069_0001
Figure imgf000070_0001
Figure imgf000071_0001
Figure imgf000072_0001
Figure imgf000073_0001
[0147] In some embodiments, the present disclosure provides a compound selected from Table 3, or a pharmaceutically acceptable salt thereof:
Table 3
Figure imgf000074_0001
Figure imgf000075_0002
In some embodiments, a compound provided herein is an agonist, and is selected from Table 3. [0148] In some embodiments, the present disclosure provides a compound selected from Table 4, or a pharmaceutically acceptable salt thereof:
Figure imgf000075_0001
Figure imgf000076_0001
In some embodiments, a compound provided herein is an antagonist, and is selected from Table
[0149] In some embodiments, the present disclosure provides a compound selected from
Table 5, or a pharmaceutically acceptable salt thereof
Table 5
Figure imgf000076_0002
Figure imgf000077_0002
In some embodiments, a compound provided herein is an antagonist, and is selected from Table 5.
[0150] In some embodiments, provided compounds are provided and/or utilized in a salt form (e g., a pharmaceutically acceptable salt form). Reference to a compound provided herein is understood to include reference to salts thereof, unless otherwise indicated.
[0151] It will be appreciated that throughout the present disclosure, unless otherwise indicated, reference to a compound of Formula I is intended to also include Formulae I- VII, and compound species of such formulas disclosed herein.
Preparing Provided Compounds
[0152] Provided compounds may generally be made by the processes described in the ensuing schemes and examples. In some embodiments, provided compounds are prepared according to Schemes 1 -4:
Figure imgf000077_0001
Scheme 1 [0153] In some embodiments, compounds described herein are prepared via Pictet-Spengler reaction between an appropriately substituted tryptamine (INT 1.1) and a heterocyclic aldehyde of interest (INT 1.2), wherein R1, R2, R3, and A are as defined in classes and subclasses herein with respect to Formula I both singly and in combination, and X is -CN, -CH2OH, -CChMe, - CH2CI, or -CI hBr. In some embodiments, INT 1.3 then undergoes selective reduction (when X is -CN), selective oxidation (when X is -CH2OH), or a two-step reduction-oxidation sequence (when X is -CChMe). In some embodiments, a B group comprising an amine and substituted with (R4)n is coupled with INT 1.4 by reductive amination to provide INT 1.5 (wherein B, R4, and n are as defined in classes and subclasses herein with respect to Formula I both singly and in combination). Alternatively, INT 1.3 undergoes a substitution (when X is -CH2CI, or CFFBr) with a B group comprising an amine and substituted with (R4)n to provide INT 1.5 directly.
[0154] As another alternative, in some embodiments, the X group of INT 1.3 is an amine precursor such as -CN or -CH2NHB0C. In some such embodiments, after conversion of the amine precursor to an amine by selective reduction or acidic deprotection, the intermediates are subjected to reductive amination to provide INT 1.5.
Figure imgf000078_0001
Scheme 2
[0155] In some embodiments, target compounds are prepared via a late-stage, two-step conversion of advanced intermediate carboxylic acids to amines. For example, in some embodiments, a compound INT 2.1 is saponified to provide carboxylic acid INT 2.2, wherein R1, R2, R3, and A are as defined in classes and subclasses herein with respect to Formula I. In some embodiments, INT 2.2 is subjected to amidation with amines of interest (e.g., a B group comprising an amine group and substituted with (R4)n), followed by amide reduction to provide compound INT 1.5.
Figure imgf000079_0001
[0156] In some embodiments, preparation of targeted compounds begins with reductive amination between appropriately substituted heterocyclic aldehydes (INT 1.2) and amines of interest (e.g., a B group comprising an amine group and substituted with (R4)n), to provide INT 3.1. In some embodiments, INT 3.1 undergoes selective reduction (when X is -CN), selective oxidation (when X is -CH2OH), a two-step reduction-oxidation sequence (when X is -CCbMe), or formylation (when X is H) to provide INT 3.2. In some embodiments, INT 3.2 is reacted with INT 1.1 in the presence of acid and heat to undergo a Pictet-Spengler reaction to provide INT 1.5.
Figure imgf000079_0002
Scheme 4
[0157] In some embodiments, compounds described herein are prepared as shown in Scheme 4. In some embodiments, INT 1.3 undergoes a Suzuki -type cross coupling reaction (when X is halogen) with INT 4.1, wherein Y is boronate ester or boronic acid and L is a covalent bond or contains at least one carbon atom (e g., L is -CH2-). In some embodiments, INT 4.2 undergoes reduction with LiAlHr to give INT 4.3, wherein R4 is Me, or acidic deprotection, followed by further functionalization, for example, via reductive amination, SN2 alkylation, or various acylation reactions to give INT 4.3, wherein R4 is alkyl.
Uses, Formulation, and Administration
[0158] The present disclosure provides uses for compounds and compositions described herein. In some embodiments, provided compounds and compositions are useful in medicine (e.g., as therapy). In some embodiments, provided compounds and compositions are useful in research as, for example, analytical tools and/or control compounds in biological assays.
Pharmaceutically Acceptable Compositions
[0159] In some embodiments, the present disclosure provides a composition comprising a compound described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. In some embodiments, the amount of compound in compositions described herein is such that it is effective to measurably induce degradation of a target in a biological sample or in a patient. In some embodiments, a composition described herein is formulated for administration to a patient in need of such composition. In some embodiments, a composition described herein is formulated for oral administration to a patient.
[0160] Pharmaceutical compositions typically contain an active agent (e.g., a compound described herein) in an amount effective to achieve a desired therapeutic effect while avoiding or minimizing adverse side effects. In some embodiments, provided pharmaceutical compositions comprise a compound described herein and one or more fillers, disintegrants, lubricants, glidants, anti-adherents, and/or anti-statics, etc. Provided pharmaceutical compositions can be in a variety of forms including oral dosage forms, topical creams, topical patches, iontophoresis forms, suppository, nasal spray and/or inhaler, eye drops, intraocular injections forms, deport forms, as well as injectable and infusible solutions. Methods of preparing pharmaceutical compositions are well known in the art.
[0161] In some embodiments, provided compounds are formulated in a unit dosage form for ease of administration and uniformity of dosage. A unit dosage form may be, for example, a liquid pharmaceutical composition containing a predetermined quantity of one or more active agents, a solid pharmaceutical composition (e.g., a tablet, a capsule, or the like) containing a predetermined quantity of one or more active agents, a sustained release formulation containing a predetermined quantity of one or more active agents, or a drug delivery device containing a predetermined amount of one or more active agents, etc.
[0162] Provided compositions may be administered using any amount and any route of administration effective for treating or lessening the severity of any disease or disorder described herein. For example, compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, intraperitoneally, intraci sternally or via an implanted reservoir. In some embodiments, the compositions are administered orally, intraperitoneally or intravenously.
[0163] Sterile injectable forms of the compositions described herein may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3 -butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.
[0164] For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
[0165] Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. [0166] In order to prolong the effect of a compound of the present disclosure, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactidepolyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
[0167] In some embodiments, provided pharmaceutically acceptable compositions are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions described herein are administered without food. In other embodiments, pharmaceutically acceptable compositions described herein are administered with food. Pharmaceutically acceptable compositions described herein may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
[0168] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fdlers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and/or i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.
[0169] Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
[0170] The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. [0171] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
[0172] Alternatively, pharmaceutically acceptable compositions described herein may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.
[0173] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
[0174] Pharmaceutically acceptable compositions described herein may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
[0175] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.
[0176] For topical applications, provided pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of compounds described herein include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
[0177] For ophthalmic use, provided pharmaceutically acceptable compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.
[0178] Pharmaceutically acceptable compositions described herein may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well- known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
[0179] Dosage forms for topical or transdermal administration of a compound disclosed herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this disclosure. Additionally, the present disclosure contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
Estrogen Receptor-Associated Diseases and Disorders
[0180] The estrogen receptor (“ER”) is involved in a variety of biological processes, relating, for example, to development of the female reproductive system, maintenance of bone mass, protection of cardiovascular and/or central nervous system components, etc. (see, for example, Pearce & Jordan Crit. Rev. Onc/Hem 50:3, 2004; Heldring Phys. Rev. 87:905, 2007). The ER has been implicated in a variety of cancers. In many tumors that express the estrogen receptor (i.e., ER+ tumors), active ERa signaling has been demonstrated to drive cell proliferation (although ERp signaling has been reported to be able to achieve tumor suppressor effects; see, for example, Nilsson & Gustafson Clin. Pharmacol. Ther. 89:44, 2011). Typically, tumors (e.g., breast tumors) with as few as 1% of cells staining positive for ER are classified as “ER+”. Therapies targeting the ER are standard of care for many patients with ER+ tumors (see, for example, Cardoso et al Annals One. <https://doi.org/10.1093/announc/mdmx036>, 2017; Rugo et al. J. Clin. Oncol. 34:3069, 2016; Senkus et al Annal One. 26:v8, 2015; Sareddy & Vadlamudi Clin. J Nat. Med, 13:801, 2015). For early stage breast cancer patients, for example, recommended therapy typically involves tumor resection, followed by ER-targeted therapy (e.g., as discussed below). For advanced breast cancer, including metastatic breast cancer, ER- targeted therapy is the mainstay.
[0181] Given the importance of ER signaling in many cancers, as well as in certain cardiovascular, inflammatory, and neurodegenerative diseases, significant effort has been invested in developing therapeutic agents and modalities that target the ER. There is some fluidity/flexibility in terminology that has been used to describe ER-targeting agents, but a variety of agents, with different mechanisms, have been developed and/or studied.
[0182] For example, some ER-targeting agents are designed and/or documented to reduce levels of estrogen (i.e., 17(3 estradiol) production. In other embodiments, some ER-targeting agents are designed and/or documented to increase levels of estrogen production.
[0183] Some ER-targeting agents are designed and/or documented to bind directly to the ER; in some cases, such agents compete with estrogen for binding to the ER and/or interfere with the allosteric changes that estrogen binding would naturally produce. Often, the term “antiestrogen” is used to refer to agents that bind to the ER, and sometimes is specifically used to indicate those agents that compete with estrogen for ER binding.
[0184] The term “selective estrogen receptor modulator, “SERM”, has been used to refer to compounds that are designed and/or documented to alter some aspect of ER activity. Some writings refer to “SERMs” as representing a particular type of anti-estrogens; other writings, however, use the term “SERM” more generally, to refer to a compound that specifically impacts some feature of ER (particularly ERa) expression and/or activity. [0185] The term “selective estrogen receptor degrader” (“SERD”) has been used to refer to compounds that are designed and/or documented to trigger or enhance degradation of the ER. In many instances, if presence of a compound correlates with reduced level of ER, the compound may be referred to as a SERD. Some writings classify compounds either as SERMs or as SERDs; others refer to SERDs as a particular type, or species, of compounds that are SERMs.
[0186] Regardless of mechanism of action of a particular agent, clinical experience thus far has revealed that incomplete effects (e.g., within an individual patient and/or across patient populations) and/or development of resistance remain a problem.
[0187] Among other things, presence or development of certain ER mutations has been reported to impact effectiveness of various ER-targeted therapies (see, for example, Jeselsohn et al Nature Rev. Clin. One. 12, 573, 2015; Gelsomino et al. Breast Cancer Res. Treat 157:253, 2016; Toy et al. 2013). Some particularly problematic mutations are those that “activate” one or more aspects of ER expression and/or function; some activating mutations have been reported that can render the ER ligand-independent (i.e., constitutively active). For example, particular mutations in the ER ligand binding domain, including D538G and Y537S, have been demonstrated to constitutively activate the ER; other mutations including deletions and/or fusions that remove the ligand binding domain, can have similar effects (see, for example, Li et al. Cell Repts 4:1116, 2013; Veeraraghavan et al Breast Cancer Research and Treatment 158, 219-232, 2016; Veeraraghavan, et al. Nature Comms 5:4577, 2014). Some reports have indicated that as many as 50% of women with metastatic breast cancer may have activating ER mutations detectible in circulating tumor DNA.
Estrogen Receptor Antagonists
[0188] In some embodiments, compounds provided herein are estrogen receptor antagonists. As used herein, an “estrogen receptor antagonist” refers to a compound or composition that produces an antagonistic effect when contacting the estrogen receptor of a subject or biological sample. In some embodiments, an estrogen receptor antagonist is characterized by having:
1. (i) between 10% and 80% increase in the E2-normalized signal in the AP assay (agonist mode) of Example 209 and (ii) between 10% and 80% reduction in the E2- normalized signal in the AP assay (antagonist mode) of Example 209; 2. (i) a pICso greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode) of Example 209; or
3. (i) a pICso greater than 7.5 and at least a 10% reduction in the E2 -normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode) of Example 209.
[0189] In some embodiments, compounds provided herein are complete estrogen receptor antagonists. As described herein, a complete estrogen receptor antagonist (a “CERAN”) is one that (1) inhibits both AF1 and AF2, and in particular inhibits AF1 activity that remains present in constitutively active ER mutants; (2) promotes ER degradation; and (3) lacks the partial ER agonist activity observed with certain other agents. The present disclosure further appreciates that many previous therapies, including for example, ARN-810, AZD9496, tamoxifen, and others, are less effective than CERANs at least in part because they only partially antagonize ER, and specifically because they inhibit activation of AF2 but not AF1. In some embodiments, an estrogen receptor antagonist is characterized by having (i) a pICso greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode) of Example 209. In some embodiments, an estrogen receptor antagonist is characterized by having (i) a pICso greater than 7.5 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209; and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode) of Example 209.
Estrogen Receptor Agonists
[0190] In some embodiments, compounds provided herein are estrogen receptor agonists. As used herein, an “estrogen receptor agonist” refers to a compound or composition that produces an agonistic effect when contacting the estrogen receptor of a subject or biological sample. In some embodiments, an estrogen receptor agonist is characterized by having (i) at least 80% increase in the E2 -normalized signal in the AP assay (agonist mode) of Example 209 and (ii) no more than 80% reduction in the E2-normalized signal in the AP assay (antagonist mode) of Example 209. Diseases, Disorders, and Conditions
[0191] The present disclosure encompasses the insight that provided compounds have a number of uses, including treatment of an ER-associated disorder (e.g., an ER-associated cancer such as breast cancer, osteoporosis, or menopause symptoms), detection, and/or characterization of certain tumors. In some embodiments, a disease, disorder, or condition is a cancer. In some embodiments, a disease, disorder, or condition is associated with a mutation in an estrogen receptor.
[0192] In some embodiments, provided compounds are useful for treating a disorder associated with increased ER activity (e.g., an ER-associated cancer such as breast cancer). In some embodiments, provided estrogen receptor antagonists (e.g., complete estrogen receptor antagonists) are useful for treating such disorders.
[0193] In some embodiments, provided compounds are useful for treating a disorder associated with decreased ER activity (e.g., menopause-related conditions or symptoms, or osteoporosis). In some embodiments, provided estrogen receptor agonists are useful for treating such disorders. Other uses of estrogen receptor agonists exist; see, e.g., Harrison, R. F. and Bonnar, i . Pharmac. Ther., 1980, 11, 451-67.
[0194] In some embodiments, the present disclosure provides a method of treating a disorder mediated by an estrogen receptor in a subject, comprising administering to the subject a therapeutically effective amount of a compound described herein.
[0195] In some embodiments, a disorder is selected from the group consisting of breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, uterine cancer, and endometriosis. In some embodiments, a disorder is breast cancer. In some embodiments, a disorder is ovarian cancer. In some embodiments, a disorder is endometrial cancer. In some embodiments, a disorder is vaginal cancer. In some embodiments, a disorder is lung cancer. In some embodiments, a disorder is bone cancer. In some embodiments, a disorder is uterine cancer. In some embodiments, a disorder is endometriosis.
[0196] In some embodiments, the present disclosure provides a method of treating a disorder associated with a mutation of an estrogen receptor in a subject, comprising administering to the subject a therapeutically effective amount of a compound described herein. In some embodiments, such a disorder is selected from the group consisting of breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, uterine cancer, and endometriosis. In some embodiments, a disorder is breast cancer. In some embodiments, a disorder is ovarian cancer. In some embodiments, a disorder is endometrial cancer. In some embodiments, a disorder is vaginal cancer. In some embodiments, a disorder is lung cancer. In some embodiments, a disorder is bone cancer. In some embodiments, a disorder is uterine cancer. In some embodiments, a disorder is endometriosis.
[0197] In some embodiments, a method of treating a disorder in a subject described herein comprises administering to the subject a compound described herein in combination with or alternation with an anti-cancer agent. In some embodiments, an anti-cancer agent is selected from an mTOR inhibitor, a CDK4/6 inhibitor, a PI3 kinase inhibitor, an aromatase inhibitor, an antibody to or inhibitor of PD-1, PD-L1 or CTLA-4, or an antibody to or inhibitor of EGFR, PGFR, or IGFR.
[0198] In some embodiments, an anti-cancer agent is a HER2 inhibitor. In some embodiments, a HER2 inhibitor is selected from tucatinib, trastuzumab, pertuzumab, ado- trastuzumab, trastuzumab emtansine, ado-trastuzumab emtansine, trastuzumab deruxtecan pertuzumab, lapatinib, and neratinib.
[0199] In some embodiments, an anti-cancer agent is an mTOR inhibitor. In some embodiments, an mTOR inhibitor is selected from everolimus sirolimus, temsirolimus, and LY3023414.
[0200] In some embodiments, an anti-cancer agent is a CDK4/6 inhibitor. In some embodiments, a CDK4/6 inhibitor is selected from palbociclib, abemaciclib, ribociclib, lerociclib, trilaciclib, and SHR6390.
[0201] In some embodiments, an anti-cancer agent is a PI3 kinase inhibitor. In some embodiments, a PI3 kinase inhibitor is selected from perifosine, CAL101, BEZ235, XL147, XL765, GDC-0941, and IPI-145.
[0202] In some embodiments, a PI3 kinase inhibitor is a PIK3CA inhibitor. In some embodiments, a PIK3CA inhibitor is selected from alpelisib, taselisib, and LY3023414.
[0203] In some embodiments, an anti-cancer agent is an aromatase inhibitor. In some embodiments, an aromatase inhibitor is selected from aminoglutethimide, testolactone, anastrozole, letrozole, exemestane, vorozole, formestane, fadrozole, 4-hydroxyandrostenedione, 1, 4, 6-androstatrien-3, 17-dione, and 4-androstene-3, 6, 17-trione. [0204] In some embodiments, an anti-cancer agent is an antibody to or inhibitor of PD-1, PD-L1 or CTLA-4.
[0205] In some embodiments, an anti-cancer agent is an antibody to or inhibitor of EGFR, PGFR, or IGFR. In some embodiments, an anti-cancer agent is erlotinib or gefitinib.
[0206] In some embodiments, a method described herein comprises administering a compound reported herein in combination or in alternation with an estrogen receptor antagonist or a partial estrogen receptor antagonist.
[0207] In some embodiments, the present disclosure provides a method of preventing recurrence of a cancer in a subject comprising administering to the subject a compound described herein. In some embodiments, a cancer is selected from breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, and uterine cancer. In some embodiments, a compound described herein is administered as an adjunctive therapy after or instead of chemotherapy, radiation, or surgery. In some embodiments, a compound is administered after surgery. In some embodiments, a compound is administered prior to surgery. In some embodiments, a cancer is a breast cancer that has progressed in the presence of endocrine or aromatase therapy.
EXAMPLES
[0208] As described in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It will be appreciated that, although the general methods depict the synthesis of certain compounds of the present disclosure, the following general methods and other methods known to one of ordinary skill in the art can be applied to all compounds and subclasses and species of each of these compounds, as described herein.
Example 1: Methyl 3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylate.
Figure imgf000091_0001
[0209] Under nitrogen atmosphere, methyl 3-formyl-lH-indole-6-carboxylate (393.00 mg, 1.93 mmol) and acetic acid (0.23 mL, 4.03 mmol) were added sequentially to the solution of 2- fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l-amine (400 mg, 1.61 mmol) in 1,2-di chloroethane (20 mL) in microwave vial, exchanged the air in the vial with nitrogen 3 times, the reaction mixture was heated at 100 °C in microwave reactor for 4 h. After cooling to room temperature, the reaction mixture was diluted with dichloromethane and washed with saturated NaHCOs solution, H2O and brine, dried with anhydrous Na2SO4, filtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 30 % ethyl acetate in hexane) to afford the title compound (320.3 mg, 45.9% yield) as pale-yellow solid. LC-MS: m/z = 434 [M+H]+.
Example 2: 3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylic acid.
Figure imgf000092_0001
[0210] Under nitrogen atmosphere, a solution of lithium hydroxide monohydrate (19.36 mg, 0.36 mmol) inH2O (0.5 mL) was added to a solution of methyl 3-((lR,3R)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylate (50 mg, 0.12 mmol) in THF (1 mL) at room temperature, then heated at 65 °C and stirred overnight. The reaction mixture was concentrated under vacuum, diluted with H2O (6 mL), adjusted pH to 2-3 with 1 N HC1 solution, and extracted with ethyl acetate. The organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford the title compound (20.0 mg, 41.3% yield) as a pale-yellow solid. LC-MS: m/z = 420 [M+H]+. Example 3: (3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indol-6-yl)methanol.
Figure imgf000093_0001
[0211] Under nitrogen atmosphere, a 1 M solution of lithium aluminum hydride (2.08 mL, 2.08 mmol) in THF was added dropwise to a solution of methyl 3-((lR,3R)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylate (300.0 mg, 0.69 mmol) in anhydrous THF (10 mL) at 0 °C, then stirred at room temperature for 3 h. The reaction mixture was cooled to 0 °C, 0.06 mL H2O was added dropwise to the reaction mixture, followed by 0.12 mL 10% NaOH solution, then 0.24 mL H2O, and stirred at room temperature for 30 min. The mixture was then fdtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 60% ethyl acetate in hexane) to afford the title compound (268.3 mg, 95.6% yield) as paleyellow solid. XH NMR (300 MHz, DMSO-de) 8 10.77 (s, 1H), 10.62 (s, 1H), 7.68-7.63 (m, 1H), 7.41 (d, J = 7.3 Hz, 1H), 7.28 (s, 1H), 7.19 (d, J = 7.3 Hz, 1H), 7.04-6.91 (m, 2H), 6.86 (d, J = 8.5 Hz, 1H), 6.64-6.59 (m, 1H), 5.26 (s, 2H), 5.01 (t, J = 5.7 Hz, 1H), 4.55-4.50 (m, 2H), 2.81- 2.50 (m, 5H), 1.39 (d, J = 21.6 Hz, 3H), 1.25 (d, J = 21.3 Hz, 3H), 0.99 (d, J = 6.7 Hz, 3H); LC- MS: m/z = 406 [M+H]+.
Example 4: 3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde.
Figure imgf000093_0002
[0212] Under nitrogen atmosphere, (3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indol-6-yl)methanol (220.00 mg, 0.54 mmol) was dissolved in anhydrous dichloromethane (12 mL) and the reaction mixture was cooled to 0 °C. (l,l,l-Triacetoxy)-l,l-dihydro-l,2-benziodoxol-3(lH)-one (276.13 mg, 0.65 mmol) was added and the mixture was stirred at 0 °C for 30 min, then stirred at room temperature for 2 h. The mixture was diluted with dichloromethane (40 mL) and poured into saturated solutions of NaHCCh (20 mL) and saturated solutions of Na2S2Ch (20 mL). It was stirred at room temperature for 60 min. The organic layer was washed with H2O (60 mL) and brine (60 mL), dried over anhydrous Na2SO4, and filtrated, then the solvent was evaporated under reduced pressure to afford the crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 60% ethyl acetate in hexane) to afford the title compound (198.7 mg, 90.8% yield) as pale-yellow solid. LC-MS: m/z = 404 [M+H]+.
Example 5: (lR,3R)-2-(2-Fluoro-2-methylpropyl)-l-(6-((3-(fluoromethyl)azetidin-l- yl)methyl)-lH-indol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000094_0001
[0213] Under nitrogen atmosphere, the solution of 3-((lR,3R)-2-(2-fhioro-2-methylpropyl)- 3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde (30.00 mg, 0.074 mmol) in anhydrous dichloromethane (0.5 mL) was added to the reaction solution of 3- (fluoromethyl)azetidine hydrochloride (18.67 mg, 0.15 mmol) and triethylamine (0.020 mL, 0.15 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture, then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (47.27 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), and the organic phase was washed with saturated NaHCCh solution and brine. The organic layer was dried with anhydrous Na2SC>4, fdtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 10% MeOH in dichloromethane) to afford the title compound (13.60 mg, 38.4% yield) as pale-yellow solid. XH NMR (300 MHz, DMSO-de) 8 10.76 (s, 1H), 10.62 (s, 1H), 7.63 (d, J = 7.5 Hz, 1H), 7.41 (d, J = 7.3 Hz, 1H), 7.20 (d, J = 8.9 Hz, 2H), 7.03-6.90 (m, 2H), 6.83 (d, J = 8.1 Hz, 1H), 6.61 (s, 1H), 5.26 (s, 1H), 4.58 (d, J = 6.2 Hz, 1H), 4.42 (d, J = 6.2 Hz, 1H), 3.58 (s, 2H), 3.45-3.36 (m, 1H), 3.24 (t, J = 7.0 Hz, 2H), 2.94 (s, 2H), 2.78-2.53 (m, 5H), 1.40 (d, J = 21.9 Hz, 3H), 1.25 (d, J = 20.8 Hz, 3H), 1.00 (d, J = 6.7 Hz, 3H); LC-MS: m/z = 477 [M+H]+.
Example 6: (lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-l-(6-((3-methylazetidin-l- yl)methyl)-lH-indol-3-yl)-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000095_0001
[0214J Under nitrogen atmosphere, a solution of 3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde (30.00 mg, 0.074 mmol) in anhydrous di chloromethane (0.5 mL) was added to a reaction solution of 3- m ethyl azetidine hydrochloride (16 00 mg, 0.15 mmol) and triethylamine (0.020 mb, 0.15 mmol) in anhydrous di chloromethane (1 mL). Acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (47.27 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), and the organic phase was washed with saturated NaHCOs solution and brine, then dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 10% MeOH in dichloromethane) to afford the title compound (13.20 mg, 38.7% yield) as pale-yellow solid. LC-MS: m/z = 459 [M+H]-.
Example 7: 3-Fluoro-N-((3-((l R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4, 9-tetrahydro- lH-pyrido[3,4-b]indol-l-yl)-lH-indol-6-yl)methyl)-N-methylpropan-l-amine
Figure imgf000096_0001
[0215] Under nitrogen atmosphere, a solution of 3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde (25.00 mg, 0.062 mmol) in anhydrous di chloromethane (0.5 mL) was added to a reaction solution of 3- fluoro-N-methyl-propan-1 -amine (11.29 mg, 0.12 mmol) and triethylamine (0.017 mb, 0.12 mmol) in anhydrous di chloromethane (1 mL). Acetic acid (0.014 mL, 0.25 mmol) was added to the reaction mixture, then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (39.40 mg, 0.19 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SC>4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 10 % MeOH in di chloromethane) to afford the title compound (10.00 mg, 33.7% yield) as pale-yellow solid. LC-MS: m/z = 479 [M+H]+.
Example 8: N-((3-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH indol-6-yl)methyl)-N-methylpropan-l-amine.
Figure imgf000096_0002
[0216] Under nitrogen atmosphere, a solution of 3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde (25.00 mg, 0.062 mmol) in anhydrous dichloromethane (0.5 mL) was added to a solution of N- methylpropan-1 -amine (9.06 mg, 0.12 mmol) and triethylamine (0.017 mL, 0.12 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.014 mL, 0.25 mmol) was added to the reaction mixture, then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (39.40 mg, 0.19 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated Nal lCOi solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 10% MeOH in dichloromethane) to afford the title compound (10.60 mg, 37.1% yield) as pale-yellow solid. LC-MS: m/z = 461 [M+H]“.
Example 9: 3-Fluoro-N-((3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4, 9-tetrahydro- lH-pyrido[3,4-b]indol-l-yl)-lH-indol-6-yl)methyl)propan-l-amine
Figure imgf000097_0001
[0217] Under nitrogen atmosphere, a solution of 3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde (30.00 mg, 0.074 mmol) in anhydrous dichloromethane (0.5 mL) was added to a solution of 3-fluoro-l- propanamine hydrochloride (16.89 mg, 0.15 mmol) and triethylamine (0.021 mL, 0.15 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (47.27 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCOi solution and brine, the organic layer was dried with anhydrous NaiSOi, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 10% MeOH in dichloromethane) to afford the title compound (10.40 mg, 30.1% yield) as paleyellow solid. LC-MS: m/z = 465 [M+H]+. Example 10: Methyl 3- ((JS,3R)-2- (2- Fluor o- 2- methylpropyl)-3 -methyl- 2, 3, 4, 9 -tetrahydro- 1 H- pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carboxylate.
Figure imgf000098_0001
[0218] Under nitrogen atmosphere, methyl 3-formyl-lH-indazole-6-carboxylate (400.00 mg, 1.96 mmol) and acetic acid (0.28 mL, 4.90 mmol) were added sequentially to the solution of 2-fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l-amine (632.47 mg, 2.55 mmol) in 1,2-di chloroethane (20 mL) in a microwave vial, exchanged the air in the vial with nitrogen 3 times, the reaction mixture was heated at 100 °C in microwave reactor for 2 h. After cooling to room temperature, the reaction mixture was diluted with dichloromethane and washed with saturated NaHCOs solution, H2O and brine, dried with anhydrous Na2SO4, filtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0% - 30% ethyl acetate in hexane) to afford the title compound (628.00 mg, 73.8% yield) as pale-yellow solid. LC-MS: m/z = 435 [M+H]+.
Example 11 : 3-((lS,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carboxylic acid.
Figure imgf000098_0002
[0219] Under nitrogen atmosphere, a solution of lithium hydroxide monohydrate (6.18 mg, 0.15 mmol) inH2O (0.5 mL) was added to the solution of methyl 3-((lS,3R)-2-(2-fhioro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6- carboxylate (16.00 mg, 0.037 mmol) in THF (1 mL) at room temperature, then heated at 65 °C and stirred overnight. The reaction mixture was concentrated under vacuum, diluted with H2O (6 mL), adjusted to pH 2-3 with 1 N HC1 solution, extracted with ethyl acetate, the organic phase was washed with brine, dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford the title compound (6.00 mg, 38.8% yield) as pale-yellow solid. LC-MS: m/z = 421 [M+H]+.
Example 12: (3-((lS,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3, 4-b]indol-l -yl)-lH-indazol- 6-yl) methanol.
Figure imgf000099_0001
[0220] Under nitrogen atmosphere, a solution of l M lithium aluminum hydride (3.80 mL, 3.80 mmol) in THF was added dropwise to a solution of methyl 3-((lS,3R)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6- carboxylate (550.00 mg, 1.27 mmol) in anhydrous THF (20 mL) at 0 °C, then stirred at room temperature for 3 h. The reaction mixture was cooled to 0 °C, 0.12 mL H2O was added dropwise to the reaction mixture, then added 0.24 mL of 10% NaOH solution, followed by 0.48 mL of H2O, stirred at room temperature for 30 min, then filtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-60% ethyl acetate in hexane) to afford the title compound (350.00 mg, 68.0% yield) as pale-yellow solid. XH NMR (300 MHz, DMSO-d6) 5 12.67 (s, 1H), 10.60 (s, 1H), 7.57-7.51 (m, 1H), 7.45-7.37 (m, 2H), 7.18 (d, J = 8.3 Hz, 1H), 6.95 (dt, J = 14.5, 6.4 Hz, 3H), 5.36 (s, 1H), 5.21 (s, 1H), 4.56 (d, J = 4.5 Hz, 2H), 2.73 (m, 5H), 1.33 (d, J = 20.8 Hz, 3H), 1.21 (d, J = 22.4 Hz, 3H), 1.03 (d, J = 6.7 Hz, 3H); LC-MS: m/z = 407 [M+H]“.
Example 13: 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde.
Figure imgf000099_0002
[0221] Under nitrogen atmosphere, (3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazol-6-yl)methanol (350.00 mg, 0.86 mmol) was dissolved in anhydrous dichloromethane (20 mL) and the reaction mixture was cooled to 0 °C. (l,l,l-Triacetoxy)-l,l-dihydro-l,2-benziodoxol-3(lH)-one (438.23 mg, 1.03 mmol) was added and the mixture was stirred at 0 °C for 30 min, then stirred at room temperature for 2 h. The mixture was diluted with dichloromethane (40 mL) and poured into saturated solutions of NaHCCh (20 mL) and saturated solutions of Na2S2Ch (20 mL). It was stirred at room temperature for 60 min. The organic layer was washed with H2O (60 mL) and brine (60 mL), dried over anhydrous Na2SO4, and filtrated, then the solvent was evaporated under reduced pressure to afford the crude product. The crude product was purified by normal phase Combiflash chromatography (0%-60% ethyl acetate in hexane) to afford the title compound (118.68 mg, 34.1% yield) as pale-yellow solid. LC-MS: m/z = 405 [M+H]+.
Example 14: (lS,3R)-2-(2-Fluoro-2-methylpropyl)-l-(6-((3-(fluoromethyl)azetidin-l- yl)niethyl)-lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000100_0001
[0222] Under nitrogen atmosphere, the solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)- 3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (30.00 mg, 0.074 mmol) in anhydrous dichloromethane (0.5 mL) was added to the reaction solution of 3-(fluoromethyl)azetidine hydrochloride (18.63 mg, 0.15 mmol) and triethylamine (0.021 mL, 0.15 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (47.27 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in dichloromethane) to afford the title compound (14.40 mg, 40.7% yield) as pale-yellow solid. ’H NMR (300 MHz, DMSO-d6) 6 12.66 (s, 1H), 10.60 (s, 1H), 7.52 (d, J = 7.9 Hz, 1H), 7.42 (d, J = 7.0 Hz, 1H), 7.32 (s, 1H), 7.18 (d, J = 7.2 Hz, 1H), 7.03- 6.91 (m, 2H), 6.88 (d, J = 8.3 Hz, 1H), 5.37 (s, 1H), 4.59 (d, J = 6.1 Hz, 1H), 4.43 (d, J = 6.2 Hz, 1H), 3.62 (s, 2H), 3.50-3.36 (m, 1H), 3.25 (d, J = 4.7 Hz, 2H), 2.96 (s, 2H), 2.88-2.52 (m, 5H), 1.34 (d, J = 21.6 Hz, 3H), 1.21 (d, J = 21.5 Hz, 3H), 1.03 (d, J = 6.7 Hz, 3H); LC-MS: m/z = 479 [M+H]+.
Example 15: (lS,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-l-(6-((3-methylazetidin-l- yl)methyl)-lH-indazol-3-yl)-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000101_0001
[0223] Under nitrogen atmosphere, the solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)- 3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l -yl)-lH-indazole-6-carbaldehyde (30.00 mg, 0.074 mmol) in anhydrous di chloromethane (0.5 mL) was added to the reaction solution of 3 -methylazetidine hydrochloride (15.96 mg, 0.15 mmol) and triethylamine (0.021 mL, 0.15 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (47.27 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous NaiSCL, filtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in di chloromethane) to afford the title compound (13.10 mg, 38.4% yield) as pale-yellow solid. LC-MS: m/z = 460 [M+H]-. Example 16: (1 S,3R)-1 -(6-((3-ethylazetidin-l -yl)methyl)-lH-indazol-3-yl)-2-(2-fluoro-2- methyl-propyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000102_0001
[0224] Under nitrogen atmosphere, a solution of 3-((l S,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous dichloromethane (1 mL) was added to the reaction solution of 3- ethylazetidine, trifluoroacetic acid (39 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous Na2SCU, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in di chloromethane) to afford the title compound (16.28 mg, 34.8% yield) as pale-yellow solid. LC-MS: m/z = 474 [M+l]+.
Example 17: 3-Fluoro-N-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9- tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazol-6-yl)methyl)-N-methylpropan-l-amine.
Figure imgf000102_0002
[0225] Under nitrogen atmosphere, the solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-
3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (30.00 mg, 0.072 mmol) in anhydrous di chloromethane (0.5 mL) was added to the reaction solution of 3-fluoro-N-methyl-propan-l-amine (13.52 mg, 0.15 mmol) and triethylamine (0.021 mL, 0.15 mmol) in anhydrous dichloromethane (ImL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (47.16 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in di chloromethane) to afford the title compound (14.18 mg, 39.9% yield) as pale-yellow solid. LC-MS: m/z = 480 [M+H]-.
Example 18: N-((3-((l S, 3R)-2- (2-Fluoro-2-methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH indazol-6-yl)methyl)-N-methylpropan-l-amine.
Figure imgf000103_0001
[0226] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (30.00 mg, 0.072 mmol) in anhydrous di chloromethane (0.5 mL) was added to the reaction solution ofN- methylpropan- 1 -amine (10.88 mg, 0.15 mmol) and triethylamine (0.021 mb, 0.15 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (47.16 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous NaiSCh, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in dichloromethane) to afford the title compound (16.00 mg, 46.6% yield) as pale-yellow solid. LC-MS: m/z = 462 [M+H]-. Example 19: 3-Fluoro-N-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2, 3,4,9- tetrahydro- 1 H-pyrido[3,4-b]indol-l -yl)-l H-indazol-6-yl)methyl)propan-l -amine.
Figure imgf000104_0001
[0227] Under nitrogen atmosphere, the solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)- 3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (30.00 mg, 0.072 mmol) in anhydrous dichloromethane (0.5 mL) was added to a solution of 3-fluoro-l- propanamine hydrochloride (16.85 mg, 0.15 mmol) and triethylamine (0.021 mL, 0.15 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At O °C, sodium triacetoxyborohydride (47.16 mg, 0.22 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in di chloromethane) to afford the title compound (15.60 mg, 45.2% yield) as pale-yellow solid. LC-MS: m/z = 466 [M+H]+.
Example 20: (lS,3R)-2-(2-fluoro-2-methylpropyl)-l-(6-(((R)-3-fluoropyrrolidin-l-yl)methyl)- lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000104_0002
[0228] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (50.00 mg, 0.12 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of(R)-(-)-3- fluoropyrrolidine hydrochloride (31.05 mg, 0.25 mmol) and triethylamine (0.035 mL, 0.25 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.028 mL, 0.49 mmol) was added to the reaction mixture then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (52.40 mg, 0.25 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in di chloromethane) to afford the title compound (22.55 mg, 38.2% yield) as pale-yellow solid. LC-MS: m/z = 478 [M+H]-.
Example 21: (lS,3R)-2-(2-fluoro-2-methylpropyl)-l-(6-(((S)-3-fluoropyrrolidin-l-yl)methyl)- lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000105_0001
[0229] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (50.00 mg, 0.12 mmol) in anhydrous dichloromethane (1 mL) was added to a solution of (3S)-(+)-3- fluoropyrrolidine hydrochloride (31.05 mg, 0.25 mmol) and triethylamine (0.035 mL, 0.25 mmol) in anhydrous di chloromethane (2 mL), then acetic acid (0.028 mL, 0.49 mmol) was added to the reaction mixture, then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (52.40 mg, 0.25 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in di chloromethane) to afford the title compound (24.86 mg, 42.1% yield) as pale-yellow solid. LC-MS: m/z = 478 [M+H]-. Example 22: (lS,3R)-l-(6-(((R)-3-ethylpyrrolidin-l-yl)methyl)-lH-indazol-3-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH-pyrido[3, 4-b]indole.
Figure imgf000106_0001
[0230] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of(R)-3- ethylpyrrolidine hydrochloride (26.83 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture, then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in di chloromethane) to afford the title compound (14.00 mg, 29.0% yield) as pale-yellow solid. LC-MS: m/z = 488 [M+H]+.
Example 23: (lS,3R)-l-(6-(((S)-3-ethylpyrrolidin-l-yl)methyl)-lH-indazol-3-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH-pyrido[3, 4-b]indole.
Figure imgf000106_0002
[0231] Under nitrogen atmosphere, the solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)- 3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous dichloromethane (1 mL) was added to the reaction solution of (S)-3-ethylpyrrolidine hydrochloride (26.83 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous di chloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SO4, fdtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in di chloromethane) to afford the title compound (20.00 mg, 41.5% yield) as pale-yellow solid. LC-MS: m/z = 488 [M+H]-.
Example 24: (lS,3R)-l-(6-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)-lH-indazol-3-yl)-2-(2- fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000107_0001
[0232] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous dichloromethane (1 mL) was added to a solution of 3- azabicyclo[3.1.0]hexane hydrochloride (23.65 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous Na2SO4, fdtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in di chloromethane) to afford the title compound (15.00 mg, 32.2% yield) as pale-yellow solid. LC-MS: m/z = 472 [M+H]-. Example 25: (1 S,3R)-2-(2-fluoro-2-methylpropyl)-l -(6-(((R)-3-(fluoromethyl)pyrrolidin-l - yl)methyl)-lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000108_0001
[0233] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of(R)-3- (fluoromethyl)pyrrolidine hydrochloride (27.61 mg, 0.20 mmol) and triethylamine (0.028 mb, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SC>4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in di chloromethane) to afford the title compound (23.00 mg, 47.3% yield) as pale-yellow solid. LC-MS: m/z = 492 [M+H]-.
Example 26: (lS,3R)-2-(2-fluoro-2-methylpropyl)-l-(6-(((S)-3-(fluoromethyl)pyrrolidin-l- yl)methyl)-lH-indazol-3-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000108_0002
[0234] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of(S)-3- (fluoromethyl)pyrrolidine hydrochloride (27.61 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture, then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated Nal lCOs solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in dichloromethane) to afford the title compound (15.16 mg, 31.2% yield) as pale-yellow solid. LC-MS: m/z = 492 [M+H]“.
Example 27: (lS,3R)-l-(6-((2-azaspiro[3.3]heptan-2-yl)methyl)-lH-indazol-3-yl)-2-(2-fluoro- 2-methylpropyl)-3-methyl-2,3, 4, 9-tetrahydro-lH-pyrido[3, 4-b]indole.
Figure imgf000109_0001
[0235] Under nitrogen atmosphere, a solution of 3-((l S,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of 2- azaspiro[3.3]heptane hydrochloride (26.43 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SC>4, fdtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in dichloromethane) to afford the title compound (16.18 mg, 33.7% yield) as pale yellow solid. LC-MS: m/z = 486 [M+l]+. Example 28: (lS,3R)-l-(6-((6,6-difluoro-2-azaspiro[3.3]heptan-2-yl)methyl)-lH-indazol-3-yl)-
2- (2-fluoro-2-methylpropyl)-3-methyl-2, 3, 4, 9-tetrahydro-lH-pyrido[3, 4-b]indole.
Figure imgf000110_0001
[0236] Under nitrogen atmosphere, a solution of 3-((l S,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous dichloromethane (1 mL) was added to a solution of 6,6-difluoro-2- azaspiro[3.3]heptane 2,2,2-trifluoroacetate (48.88 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in dichloromethane) to afford the title compound (22.16 mg, 43.0% yield) as pale yellow solid. LC-MS: m/z = 522 [M+l]+.
Example 29: 6-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH pyrido-[3,4-b]indol-l-yl)-lH-indazol-6-yl)methyl)-2-oxa-6-azaspiro[3.3]heptane.
Figure imgf000110_0002
[0237] Under nitrogen atmosphere, a solution of 3-((l S,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (40.00 mg, 0.099 mmol) in anhydrous dichloromethane (1 mL) was added to the reaction solution of 2-oxa- 6-azaspiro[3.3]heptane oxalate (37.42 mg, 0.20 mmol) and triethylamine (0.028 mL, 0.20 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.023 mL, 0.40 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (41.92 mg, 0.20 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous Na2SO4, fdtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in di chloromethane) to afford the title compound (19.56 mg, 40.6% yield) as pale yellow solid. LC-MS: m/z = 488 [M+l]+.
Example 30: 2-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]-indol-l-yl)-lH-indazol-6-yl)methyl)-2-azaspiro[3.3]heptan-6-ol.
Figure imgf000111_0001
[0238] Under nitrogen atmosphere, a solution of 3-((l S,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (30.00 mg, 0.072 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of 2- azaspiro[3.3]heptan-6-ol hydrochloride (22.19 mg, 0.15 mmol) and triethylamine (0.020 mL, 0.15 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (31.44 mg, 0.15 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCOs solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in dichloromethane) to afford the title compound (5.00 mg, 13.4% yield) as pale yellow solid. LC-MS: m/z = 502[M+l]+. Example 31: l-((3-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-LE[- pyrido-[3,4-b]indol-l-yl)-lE[-indazol-6-yl)methyl)azetidine-3-carbonitrile.
Figure imgf000112_0001
[0239] Under nitrogen atmosphere, a solution of 3-((l S,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (30.00 mg, 0.072 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of azetidine-3- carbonitrile hydrochloride (17.59 mg, 0.15 mmol) and triethylamine (0.020 mL, 0 15 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.017 mL, 0.30 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (31.44 mg, 0.15 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%- 10 % MeOH in dichloromethane) to afford the title compound (12.36 mg, 35.4% yield) as pale yellow solid. LC-MS: m/z = 471[M+1]+.
Example 32: (lS,3R)-l-(6-((6-fluoro-2-azaspiro[3.3]heptan-2-yl)methyl)-lH-indazol-3-yl)-2- (2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole.
Figure imgf000112_0002
[0240] Under nitrogen atmosphere, a solution of 3-((l S,3R)-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde (20.00 mg, 0.049 mmol) in anhydrous dichloromethane (1 mL) was added to a solution of 6-fluoro-2- azaspiro[3.3]heptane 2,2,2-trifluoroacetate (22.66 mg, 0.099 mmol) and triethylamine (0.014 mL, 0.099 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.011 mL, 0.20 mmol) was added to the reaction mixture then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (20.96 mg, 0.099 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCO3 solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10 % MeOH in dichloromethane) to afford the title compound (13.56 mg, 54.5% yield) as pale yellow solid. LC-MS: m/z = 504[M+1]+. Example 33: (1S,3R)-1-(6-((6,6-difluoro-1-azaspiro[3.3]heptan-1-yl)methyl)-1H-indazol-3-yl)- 2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole. [0241] Under nitrogen
Figure imgf000113_0001
-2-(2-fluoro-2-methylpropyl)-3- methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)-1H-indazole-6-carbaldehyde (20.00 mg, 0.049 mmol) in anhydrous dichloromethane (1 mL) was added to a solution of 6,6-difluoro-1- azaspiro[3.3]heptane oxalate (17.62 mg, 0.049 mmol) and triethylamine (0.014 mL, 0.099 mmol) in anhydrous dichloromethane (2 mL), then acetic acid (0.011 mL, 0.20 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (20.96 mg, 0.099 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCO3 solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-10% MeOH in dichloromethane) to afford the title compound (6.32 mg, 24.5% yield) as pale yellow solid. LC-MS: m/z = 522[M+1]+.
Example 34: Methyl 3-((lR,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carboxylate.
Figure imgf000114_0002
[0242] Under nitrogen atmosphere, methyl 3-formyl-lH-indole-6-carboxylate (325.10 mg, 1.60 mmol) and acetic acid (0.16 mL, 2.86 mmol) were added sequentially to a solution of (R)-N- ( 1 -( 1 H-indol-3 -yl)propan-2-yl)-3 -((tert-butyl diphenyl silyl)oxy)-2,2-difluoropropan- 1 -amine (506.70 mg, 1.00 mmol) in anhydrous toluene (12 mL) in microwave vial, purged with nitrogen 3 times, the reaction mixture was heated at 130 °C in microwave reactor for 2 h. After cooling to room temperature, the reaction mixture was concentrated under vacuum and extracted with ethyl acetate and H2O, washed with saturated Nal ICCh solution, H2O, and brine, dried with anhydrous Na2SC>4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combifl ash chromatography (0%-30% ethyl acetate in hexane) to afford the title compound (0.21 g, 30.4% yield) as pale-yellow solid. LC-MS: m/z = 692 [M+H]+.
Example 35: (3-((lR,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lEl-pyrido[3,4-b]indol-l-yl)-lE[-indol-6-yl)methanol.
Figure imgf000114_0001
[0243] Under nitrogen atmosphere, a 1 M solution of lithium aluminum hydride (0.87 mL,
0.87 mmol) in THF was added dropwise to a solution of methyl 3-((lR,3R)-2-(3-((tert- butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol- l-yl)-lH-indole-6-carboxylate (200.00 mg, 0.29 mmol) in anhydrous THF (6 mL) at 0 °C, then stirred at room temperature for 3 h. The reaction mixture was cooled to 0 °C, 0.12 mL H2O was dropwise to the reaction mixture, then added 0.24 mL 10% NaOH solution, followed by 0.48 mL H2O, stirred at room temperature for 30 min, then fdtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-60% ethyl acetate in hexane) to afford the title compound (0.12 g, 62.5% yield) as pale-yellow solid. LC-MS: m/z = 664[M+H]+.
Example 36: 3-((lR,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indole-6-carbaldehyde.
Figure imgf000115_0001
[0244] Under nitrogen atmosphere, (3-((lR,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2- difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indol-6- yl)methanol (120.00 mg, 0.18 mmol) was dissolved in anhydrous dichloromethane (4 mL) and the reaction mixture was cooled to 0 °C. (l,l,l-Triacetoxy)-l,l-dihydro-l,2-benziodoxol-3(lH)- one (92.00 mg, 0.22 mmol) was added and the mixture was stirred at 0 °C for 30 min, then stirred at room temperature for 2 h. The mixture was diluted with dichloromethane (40 mL) and poured into saturated solutions of NaHCOs (20 mL) and saturated solutions of NaiSiCh (20 mL). It was stirred at room temperature for 60 min. The organic layer was washed with H2O (60 mL) and brine (60 mL), dried over anhydrous Na2SO4, and filtrated, then the solvent was evaporated under reduced pressure to afford the crude product. The crude product was purified by normal phase Combiflash chromatography (0%-60% ethyl acetate in hexane) to afford the title compound (80.00 mg, 66.9% yield) as pale-yellow solid. LC-MS: m/z = 662 [M+H]+. Example 37: 2,2-difluoro-3-((lR,3R)-l-(6-((3-(fluoromethyl)azetidin-l-yl)methyl)-lH-indol-3- yl)-3-methyl-l,3,4,9-tetrahydro-2H-pyrido[3,4-b]indol-2-yl)propan-l-ol.
Figure imgf000116_0001
[0245] Under nitrogen atmosphere, a solution of 3-((lR,3R)-2-(3-((tert- butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol- l-yl)-lH-indole-6-carbaldehyde (60.00 mg, 0.091 mmol) in anhydrous dichloromethane (1 mL) was added to the reaction solution of 3-(fluoromethyl)azetidine hydrochloride (22.77 mg, 0.18 mmol) and triethylamine (0.025 mL, 0.18 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.021 mL, 0.36 mmol) was added to the reaction mixture then stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (38.43 mg, 0.18 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with di chloromethane (10 mL), the organic phase was washed with saturated NaHCCh solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product, which was used for the subsequent reaction without further purification.
[0246] Under nitrogen atmosphere, a 1 M solution of tetrabutylammonium fluoride (0.18 mL, 0.18 mmol) in THF was added to a solution of crude product described above in anhydrous THF (4 mL) at room temperature, then stirred overnight. The reaction mixture was quenched with H2O, extracted with ethyl acetate, the organic phase was washed with saturated H2O and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-20% MeOH in di chloromethane) to afford the title compound (6.50 mg, 14.4% yield) as pale-yellow solid. LC-MS: m/z = 497 [M+H]+. Example 38: Methyl 3-((lS,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3- methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carboxylate.
Figure imgf000117_0001
[0247] Under nitrogen atmosphere, methyl 3 -formyl- lH-indazole-6-carboxylate (193.42 mg,
0.95 mmol) and acetic acid (0.13 mL, 2.26 mmol) were added sequentially to a solution of (R)- N-(l-(lH-indol-3-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-l-amine
(400.00 mg, 0.79 mmol) in anhydrous toluene (10 mL) in microwave vial, exchanged the air in the vial with nitrogen 3 times, the reaction mixture was heated at 90 °C overnight. After cooling to room temperature, the reaction mixture was concentrated under vacuum and extracted with ethyl acetate and H2O, washed with saturated NaHCOs solution, H2O and brine, dried with anhydrous Na2SO4, filtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-30% ethyl acetate in hexane) to afford the title compound (0.50 g, 91.4% yield) as pale-yellow solid. LC-MS: m/z = 693 [M+H]+.
Example 39: (3-((lS,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazol-6-yl)methanol.
Figure imgf000117_0002
[0248] Under nitrogen atmosphere, a 1 M solution of lithium aluminum hydride (2.16 mL, 2.16 mmol) in THF was added dr op wise to the solution of methyl 3-((lS,3R)-2-(3-((tert- butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol- l-yl)-lH-indazole-6-carboxylate (500.00 mg, 0.72 mmol) in anhydrous THF (12 mL) at 0 °C, then stirred at room temperature for 3 h. The reaction mixture was cooled to 0 °C, 0.40 mb H2O was dropwise to the reaction mixture, then added 0.80 mb 10% NaOH solution, then added 1.20 mb H2O, stirred at room temperature for 30 min, then filtered and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-60% ethyl acetate in hexane) to afford the title compound (0.45 g, 93.8% yield) as pale-yellow solid. LC-MS: m/z = 665[M+H]+.
Example 40: 3-((lS,3R)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl- 2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazole-6-carbaldehyde.
Figure imgf000118_0001
[0249] Under nitrogen atmosphere, (3-((lS,37?)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2- difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol-l-yl)-lH-indazol-6- yl)methanol (0.45g, 0.68 mmol) was dissolved in anhydrous dichloromethane (16 mb) and manganese dioxide (2.35 g, 27.07 mmol) was added, and the mixture was stirred at room temperature for 24 h. The mixture was filtered, then the solvent was evaporated under reduced pressure to afford the crude product. The crude product was purified by normal phase Combiflash chromatography (0%-50% ethyl acetate in hexane) to afford the title compound (0.21 g, 46.8% yield) as pale-yellow solid. LC-MS: m/z = 663 [M+H]+.
Example 41: 2,2-difluoro-3-((lS,3R)-l-(6-((3-(fluoromethyl)azetidin-l-yl)methyl)-lH-indazol- 3-yl)-3-methyl-l,3,4,9-tetrahydro-2H-pyrido[3,4-b]indol-2-yl)propan-l-ol.
Figure imgf000118_0002
[0250] Under nitrogen atmosphere, a solution of 3-((lS,3R)-2-(3-((tert- butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indol- l-yl)-lH-indazole-6-carbaldehyde (60.09 mg, 0.097 mmol) in anhydrous di chloromethane (1 mL) was added to a solution of 3-(fluoromethyl)azetidine hydrochloride (22.77 mg, 0.18 mmol) and triethylamine (0.025 mL, 0.18 mmol) in anhydrous dichloromethane (1 mL), then acetic acid (0.021 mL, 0.36 mmol) was added to the reaction mixture and stirred at room temperature for 1 h. At 0 °C, sodium triacetoxyborohydride (38.43 mg, 0.18 mmol) was added to the reaction mixture, then stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (0.1 mL), diluted with dichloromethane (10 mL), the organic phase was washed with saturated NaHCOi solution and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product, which was used for the following reaction without further purification.
[0251] Under nitrogen atmosphere, a 1 M solution of tetrabutylammonium fluoride (0.18 mL, 0.18 mmol) in THF was added to the solution of crude product described above in anhydrous THF (4 mL) at room temperature, then stirred overnight. The reaction mixture was quenched with H2O, extracted with ethyl acetate, the organic phase was washed with saturated H2O and brine, the organic layer was dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford crude product. The crude product was purified by normal phase Combiflash chromatography (0%-20% MeOH in di chloromethane) to afford the title compound (14.00 mg, 31.0% yield) as pale-yellow solid. LC-MS: m/z = 498 [M+H]-.
Figure imgf000119_0001
[0252] In DMF (0.5 mL) was dissolved 7-fluoro-lH-indole-6-carbaldehyde (100 mg, 0.61 mmol) and acetic acid (70 uL, 1.22 mmol). To this was added 3-(fluoromethyl)azetidine hydrochloride (92 mg, 1.23 mmol) dissolved in DMF (0.5 mL) and TEA (170 pL, 1.23 mmol) and stirred for 30 min. Sodium triacetoxyborohydride (390 mg, 0.1.84 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 N NaOH solution and extracted with EtOAc. The combined organic layers were washed with brine solution five times, dried over sodium sulfate, filtered, and dried in vacuo to give the title product (140 mg, 97% yield).
Figure imgf000120_0001
[0253] 7 -Fluoro-6-[[3-(fluoromethyl)azetidin-l -yl]methyl]-lH-indole (115 mg, 0.49 mmol) and hexamethylenetetramine (218 mg, 1.56 mmol) were dissolved in 3 mL ethanol, acetic acid (128 uL, 2.24 mmol) was added and stirred at 100 °C for 6 hours. The reaction was quenched with saturated sodium bicarbonate solution and extracted with EtOAc. The combined organics were dried over sodium sulfate, fdtered, and dried in vacuo. The crude was purified via flash chromatography using a 24 g silica column in a 0-15% MeOH in DCM gradient to give the title compound (30 mg, 23% yield). LCMS: m/z = 265.3 [M+H]+.
Example 44: (lR,3R)-l-[7-Fluoro-6-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-indol-3-yl]-2- (2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000120_0002
[0254] 7-Fluoro-6-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-indole-3-carbaldehyde (30 mg, 0.11 mmol) and 2-fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l- amine (56 mg, 0.22 mmol) were dissolved in 10 mL of dioxane, acetic acid (26 uL, 0.45 mmol) was added and stirred at 120 °C for 4 hours. The reaction crude was neutralized with saturated sodium bicarbonate solution and the organic layer was separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine solution. It was then dried over sodium sulfate, filtered, and dried in vacuo. The crude was first purified via flash chromatography on an 11 g Biotage™ Sfar KP-Amino column in 0-15% IPA in DCM with 1% TEA as modifier. The resulting material was dried, then dissolved in 1.5 mL DMA and purified by HPLC on Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-50% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white, fluffy, solid (1.0 mg, 1.8% yield). LCMS: m/z = 495.5 [M+H]+.
Example 45: (lR,3R)-l-(6-bromo-lH-indol-3-yl)-2-(2-fluoro-2-methylpropyl)-3-methyl-
Figure imgf000121_0001
[0255] A solution of 6-bromo-lH-indazole-3-carbaldehyde (1.00 equiv, 338 mg, 1.50 mmol), 2-fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l-amine (1.20 equiv, 447 mg, 1.80 mmol) and acetic acid (4.00 equiv, 0.34 mL, 6.00 mmol) in DCE (4 mL) was heated at 100 °C for 90 min. NaiCOs (aq, 10 mL) was added and the reaction extracted with DCM (5 mL). The solvent was removed and the reaction was purified via silica gel (40 g, Gold) eluting at 25% hexanes:EtOAc to afford (3R)-l-(6-bromo-lH-indazol-3-yl)-2-(2-fluoro-2-methyl-propyl)-3- methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole (368 mg, 0.808 mmol, 53.88 % yield) as a yellow solid. LCMS: m/z = 455.1, 456.9 [M+H]+.
Example 46: tert-butyl 3-((3-((lR,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-
Figure imgf000121_0002
[0256] A solution of (3R)-l-(6-bromo-lH-indazol-3-yl)-2-(2-fluoro-2-methyl-propyl)-3- methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole (1.00 equiv, 140 mg, 0.307 mmol), potassium;(l- tert-butoxycarbonylazetidin-3-yl)methyl-trifluoro-boranuide (1.95 equiv, 166 mg, 0.599 mmol), cataCXium Pd G4 (0.200 equiv, 46 mg, 0.0615 mmol), cesium carbonate (3.00 equiv, 301 mg, 0.922 mmol) in 6: 1 toluene:water (3 mL:500 pL). The reaction was microwaved at 120 °C, 140 °C and 160 °C for 5 minutes each. Water (2 mL) was added and the reaction extracted with EtOAc (3 mL x 2). The combined organic layers were dried over sodium sulfate and purified twice via silica gel (25 g, Sfar and 28g, KP-Amino) eluting at 70% hexanes:EtOAc to afford tert-butyl 3-[[3-[(3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indazol-6-yl]methyl]azetidine-l-carboxylate (10 mg, 0.0183 mmol, 5.96 % yield) as a beige solid. LCMS: m/z = 546.0 [M+H]+.
Example 47: (lR,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-l-(6-((l-methylazetidin-3-
Figure imgf000122_0001
[0257] To a solution of lithium aluminum hydride (4.81 equiv, 0.15 mL, 0.150 mmol) in THF (1 mL) at 0 °C was added tert-butyl 3-[[3-[(3R)-2-(2-fluoro-2-methyl-propyl)-3- methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-lH-indol-6-yl]methyl]azetidine-l-carboxylate (1.00 equiv, 17 mg, 0.0312 mmol). After stirring for 4 d, sodium sulfate decahydrate was added along with THF (1 mL). After 1.5 h, water (2 mL) was added and the reaction was extracted with EtOAc (2 mL x 2). The combined organic layers were dried over sodium sulfate, concentrated and purified via HPLC eluting with water (0.1 % formic acid):ACN (0.1 % formic acid) to afford (3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l-[6-[(l-methylazetidin-3-yl)methyl]-lH- indol-3-yl]-l,3,4,9-tetrahydropyrido[3,4-b]indole (4.7 mg, 0.0102 mmol, 32.84 % yield) as an orange solid. LCMS: m/z = 459.3 [M+H]+.
Example 48: Methyl 2-[(3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-5-carboxylate.
Figure imgf000123_0001
[0258] To 2-fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l -amine (1.00 equiv, 122 mg, 0.492 mmol) and methyl 2-formyl-lH-indole-5-carboxylate (1.00 equiv, 100 mg, 0.492 mmol) in DCE (4 mL) under nitrogen was added acetic acid (2.00 equiv, 0 056 mL, 0.984 mmol) and heated at 110 °C for 3 h in the microwave. The reaction mixture was extracted with DCM, washed with saturated sodium bicarbonate, dried over sodium sulfate, filtered and concentrated. The crude was purified by silica gel chromatography using 0-40% EtOAc in hexane to give title compound (160 mg, 0.369 mmol, 74.99 % yield). LCMS: m/z = 434.5 [M+H]+.
Example 49: [2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-5-yl]methanol.
Figure imgf000123_0002
[0259] To methyl 2-[(3 R)-2-(2-fluoro-2-methyl-propyl)-3 -methyl- 1, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-5-carboxylate (1.00 equiv, 160 mg, 0.369 mmol) in THF (10 mL) at 0 °C under nitrogen was added lithium aluminum hydride (2.00 equiv, 0.74 mL, 0.738 mmol). The reaction was stirred at 0 °C for 2 h, diluted with 15 mL of MTBE and treated with 30 pL of water, followed by 30 pL of 15% aqueous NaOH and then 90 pL of water. The reaction was warmed to room temperature and stirred for 15 mins and to this was added anhydrous sodium sulfate stirred and the reaction mixture filtered through Celite®. The filtrated was concentrated and the crude was purified by silica gel chromatography using 0-50% EtOAc in hexane to give title compound (120 mg, 0.296 mmol, 80.18 % yield). LCMS: m/z = 406.3 [M+H]+.
Example 50: 2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-5-carbaldehyde.
Figure imgf000124_0001
[0260] To [2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-5-yl]methanol (1.00 equiv, 70 mg, 0.173 mmol) in anhydrous DMSO (1 mL) was added l-hydroxy-l,2-benziodoxol-3(lH)-one 1-oxide (1.05 equiv, 113 mg, 0.181 mmol) and stirred at room temperature for 2 h. The reaction was diluted with water, extracted with ethyl acetate and the layers separated. The EtOAc layer was washed with water, brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-50% EtOAc in hexane to give title compound (40 mg, 0.0991 mmol, 57.43 % yield). LCMS: m/z = 404.2 [M+H]+.
Example 51: (lR,3R)-l-[5-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-indol-2-yl]-2-(2-fluoro- 2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000124_0002
[0261] To a solution of 2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-5-carbaldehyde (1.00 equiv, 20 mg, 0.0496 mmol) in DMF (0.5 mL) was added the 3-(fluoromethyl)azetidin-l-ium; chloride (1.00 equiv, 6.2 mg, 0.0496 mmol) dissolved in DMF (0.5 mL) and triethylamine (2.00 equiv, 0.014 mL, 0.0991 mmol). To the reaction mixture was added acetic acid (2.00 equiv, 0.0057 mL, 0.0991 mmol) and stirred at room temperature for 1 h and then treated with sodium triacetoxyborohydride (3.00 equiv, 32 mg, 0.149 mmol) and stirred for 1 h. The reaction was quenched with 1 mL of methanol and the crude directly purified by C-18 reverse phase chromatography using prep HPLC, eluting with 10-50% ACN in water with 0.1% formic acid as modifier, to give title compound (6.0 mg, 0.0126 mmol, 25.40 % yield). LCMS: m/z = 477.9 [M+H]+.
Example 52: 2-(hydroxymethyl)-lH-indole-6-carbonitrile.
Figure imgf000125_0001
[0262] Methyl 6-cyano-lH-indole-2-carboxylate (1.00 equiv, 0.13 g, 0.624 mmol) was added to solution of lithium borohydride (2.00 equiv, 0.62 mL, 1 25 mmol) in THF (5mL) and water (0.5 mL). The reaction was stirred for 16 h at 25 °C, then quenched with water (1 mL) and made acidic with 1 N HC1. The reaction mixture was extracted with ethyl acetate, dried over Na2SO4, filtered, and concentrated under vacuum to give title compound (90 mg, 0.523 mmol, 83.71 % yield) which was used without further purification. LCMS: m/z = 173.2 [M+H]+.
Example 53: 2-formyl-lH-indole-6-carbonitrile.
Figure imgf000125_0002
[0263] To 2-(hydroxymethyl)-lH-indole-6-carbonitrile (1.00 equiv, 90 mg, 0.523 mmol) in anhydrous DMSO (1 mL) was added l-hydroxy-l,2-benxiodoxoL3(lH)-one 1-oxide (1.00 equiv, 325 mg, 0.523 mmol) and stirred at 25 °C for 2 h. The reaction mixture was diluted with water, extracted with ethyl acetate, and the EtOAc layer was washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude was purified by silica gel chromatography, eluting with 10-40% EtOAc in hexane, to give title compound (65 mg, 0.382 mmol, 73.08 % yield). LCMS: m/z = 171.2 [M+H]+. Example 54: 2-formyl-lH-indole-6-carbonitrile.
Figure imgf000126_0001
[0264] To 2-(hydroxymethyl)-lH-indole-6-carbonitrile (1.00 equiv, 90 mg, 0.523 mmol) in anhydrous DMSO (1 mL) was added l-hydroxy-l,2-benxiodoxol-3(lH)-one 1-oxide (1.00 equiv, 325 mg, 0.523 mmol) and stirred at 25 °C for 2 h. The reaction mixture was diluted with water, extracted with ethyl acetate, and the EtOAc layer was washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude was purified by silica gel chromatography, eluting with 10-40% EtOAc in hexane, to give title compound (65 mg, 0.382 mmol, 73.08 % yield). LCMS: m/z = 171.2 [M+H]+.
Example 55: 2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lE[-indole-6-carbaldehyde.
Figure imgf000126_0002
[0265] To 2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-6-carbonitrile (1.00 equiv, 105 mg, 0.262 mmol) in DCM (10 mL) under an atmosphere of nitrogen at -76 °C was added the diisobutylaluminum hydride (3.00 equiv, 0.79 mL, 0.787 mmol) and stirred for 2 h. The reaction was the quenched with ethyl acetate, warmed to 0 °C and then treated with saturated solution of Rochelle's salt. The reaction mixture was stirred for 30 mins, and the layers separated. The aqueous phase was extracted with EtOAc, washed with brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-30% EtOAc in hexane to give title compound (60 mg, 0.149 mmol, 56.72 % yield). 404.2 [M+H]+. Example 56: (lR,3R)-l-[6-[[3-(fluorometbyl)azetidin-l-yl]metliyl]-lH-indol-2-yl]-2-(2-fluoro- 2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000127_0001
[0266] To solution of 3-(fluoromethyl)azetidine hydrochloride (2.00 equiv, 12 mg, 0.0991 mmol) in DMF (0.5 mL) was added triethylamine (2.00 equiv, 0.014 mL, 0.0991 mmol), followed by a solution of 2-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-6-carbaldehyde (1.00 equiv, 20 mg, 0.0496 mmol) in DMF (0.5 mL). To the reaction was added acetic acid (2.00 equiv, 0.0057 mL, 0.0991 mmol) and stirred at room temperature for 1 h. The reaction mixture was treated with sodium triacetoxyborohydride (3.00 equiv, 32 mg, 0.149 mmol) and stirred for 1 h. The reaction was quenched with 1 mL of methanol and purified by C-18 reverse phase chromatography using a Phenomenex Luna column, eluting with 10-50% ACN in water with 0.1% formic acid as modifier, to give title compound (4.0 mg, 0.00839 mmol, 16.93 % yield). 477.2 [M+H]+.
Figure imgf000127_0002
[0267] To methyl 5-cyano-lH-indole-2-carboxylate (1.00 equiv, 1.00 g, 5.00 mmol) in DCM (30 mL) at 0 °C under nitrogen was added the diisobutylaluminum hydride (4.00 equiv, 20 mL, 20.0 mmol) and stirred at 0 °C for 2 h. The reaction was quenched with ethyl acetate, followed by saturated solution of Rochelle's salt, and stirred for 30 mins. The organic layer was separated and aqueous layer, extracted with additional DCM twice. The organic layer was dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-10% MeOH in DCM to give title compound (600 mg, 3.43 mmol, 68.6 % yield). LCMS: m/z = 176.1 [M+H]+. Example 58: [5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-2-yl]methanol.
Figure imgf000128_0001
[0268] To 2-(hydroxymethyl)-lH-indole-5-carbaldehyde (1.00 equiv, 200 mg, 1.14 mmol) in DCE (3 mL)/l,4-di oxane (3 mL) was added the 2-fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl- ethyl]-2-methyl-propan-l -amine (1.00 equiv, 284 mg, 1.14 mmol) and acetic acid (2.00 equiv, 0.13 mL, 2.28 mmol) and heated at 1 10 °C in microwave for 6 h. The reaction mixture was extracted with DCM, washed with saturated sodium bicarbonate solution, dried over sodium sulfate, fdtered, and concentrated. The crude was purified by silica gel chromatography using a 24 g RediSep gold column eluting with 0-10% methanol in DCM to give title compound (150 mg, 0.370 mmol, 32.40 % yield). LCMS: m/z = 406.9 [M+H]+.
Example 59: 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde
Figure imgf000128_0002
[0269] To [5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-2-yl]methanol (1.00 equiv, 150 mg, 0.370 mmol) in DMSO (2.5 mL) was added l-hydroxy-l,2-benziodoxol-3(lH)-one 1-oxide (1.00 equiv, 230 mg, 0.370 mmol) and stirred at room temperature for 1 h. The reaction was diluted with water, extracted with ethyl acetate, and the layers separated. The EtOAc layer was washed with water, brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-40% EtOAc in hexane to give title compound (130 mg, 0.322 mmol, 87.1 % yield). LCMS: m/z = 404.1 [M+H]+. Example 60: (lR,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-indol-5-yl]-2-(2-fluoro- 2-methyl-propyl)-3-methyl-L,3,4,9-tetrahydropyrido[3,4-b]indole
Figure imgf000129_0001
[0270] To solution of 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde (1.00 equiv, 60 mg, 0.149 mmol) in DMF (2 mL) was added 3-(fluoromethyl)azetidin-l-ium; chloride (2.00 equiv, 37 mg, 0 297 mmol) dissolved in DMF (2 mL) and triethylamine (2.00 equiv, 0.041 mL, 0.297 mmol). To the reaction mixture was added acetic acid (2.00 equiv, 0.017 mL, 0.297 mmol) and stirred at room temperature for 1 h and then treated with sodium triacetoxyborohydride (3.00 equiv, 95 mg, 0.446 mmol) and stirred for additional 2 h. The reaction was quenched with 1 mL of methanol and the crude directly purified by C-18 reverse phase chromatography on a Phenomenex Luna column, eluting with 10-50% ACN in water with 0.1% formic acid as modifier, to give title compound (30 mg, 0.0629 mmol, 42.3 % yield). LCMS: m/z = 477.3 [M+H]+.
Example 61: (lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l-[2-[(3-methylazetidin-l- yl)methyl]-lH-indol-5-yl]-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000129_0002
[0271] 3-Methylazetidin-l-ium chloride was used to synthesize the title compound using general procedure from Example 60 to give title compound (5.0 mg, 0.0109 mmol, 29.33 % yield). LCMS: m/z = 459.2 [M+H]+. Example 62: (1R,3R)-1 -[2-(3-azabicyclo[3.1.0]hexan-3-ylmethyl)- 1 H-indol-5-yl]-2-(2-fluoro- 2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000130_0001
[0272] 3-Azabicyclo[3.1.0]hexane hydrochloride (2.00 equiv, 8.9 mg, 0.0744 mmol) was used to synthesize the title compound using general procedure from Example 60 to give title compound (9.0 mg, 0.0191 mmol, 51.44 % yield). LCMS: m/z = 471.3 [M+H]+.
Example 63: (lR,3R)-l-[2-(2-azaspiro[3.3]heptan-2-ylmethyl)-lH-indol-5-yl]-2-(2-fluoro-2- methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000130_0002
[0273] 2-Azoniaspiro[3.3]heptane; chloride (2.00 equiv, 9.9 mg, 0.0744 mmol) was used to synthesize the title compound using the general procedure from Example 60 to give title compound (5.0 mg, 0.0103 mmol, 27.75 % yield). LCMS: m/z = 485.1 [M+H]+.
Example 64: (lR,3R)-2-(2-fluoro-2-methyl-propyl)-l-[2-[[(3S)-3-fluoropyrrolidin-l- yl]methyl]-lH-indol-5-yl]-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000130_0003
[0274] (3S)-3-Fluoropyrrolidin-l-ium; chloride (2.00 equiv, 9.3 mg, 0.0744 mmol) ) was used to synthesize the title compound using general procedure from Example 60 to give title compound (5.0 mg, 0.0105 mmol, 28.22 % yield). LCMS: m/z = 477.3 [M+H]+.
Example 65: [5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methanol.
Figure imgf000131_0002
[0275] To 2-(hydroxymethyl)-lH-indole-5-carbaldehyde (1.20 equiv, 166 mg, 0.947 mmol) in DCE (4 mL)/ 1,4-dioxane (4 mL) was added 3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-N- [(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]propan-l-amine (1.00 equiv, 400 mg, 0.789 mmol) and acetic acid (2.00 equiv, 0.090 mL, 1.58 mmol) and stirred at 110 °C in microwave for 6 h. The reaction mixture was extracted with DCM, washed with saturated sodium bicarbonate solution, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using a 24 g column eluting with 0-30% EtOAc in hexane to give title compound (200 mg, 0.301 mmol, 38.16 % yield). LCMS: m/z = 664.1 [M+H]+.
Example 66: 5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde.
Figure imgf000131_0001
[0276] To [5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methanol (1.00 equiv, 200 mg, 0.301 mmol) in DMSO (3 mL) was added the l-hydroxy-l,2-benziodoxol-3(lH)-one 1-oxide (1.00 equiv, 187 mg, 0.301 mmol) and stirred at room temperature for 2 h. The reaction was quenched with water, extracted with ethyl acetate and layers separated. The EtOAc layer was washed with water, brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-10% EtOAc in hexane to give title compound (130 mg, 0.196 mmol, 65.2 % yield). LCMS: m/z = 662.1 [M+H]+.
Example 67: 2,2-difluoro-3-[(lR,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-indol-5- yl]-3-methyl-l,3, 4, 9-tetrahydropyrido[3, 4-b]indol-2-yl]propan-l -ol
Figure imgf000132_0001
[0277] To a solution of 5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde (1.00 equiv, 50 mg, 0.0755 mmol) in DMF (0.5 mL) was added 3-(fluoromethyl)azetidine hydrochloride (2.00 equiv, 19 mg, 0.151 mmol) dissolved in DMF (0.5 mL) and triethylamine (2.00 equiv, 0.021 mL, 0.151 mmol). To the reaction mixture was added acetic acid (2.00 equiv, 0.0086 mL, 0.151 mmol) and stirred at room temperature for 1 h and then treated with sodium triacetoxyborohydride (3.00 equiv, 48 mg, 0.227 mmol) and stirred for 1 h. The reaction was quenched with 1 mL of methanol and extracted with ethyl acetate, washed with water, brine, dried over sodium sulfate, filtered, and concentrated. The crude was dissolved in THF (2 mL) and treated with tetra-N-butylammonium fluoride (2.00 equiv, 0.15 mL, 0.151 mmol) and stirred at room temp for 1 h. The reaction was extracted with 1 : 1 EtOAc/MTBE, washed with water and concentrated. The crude was purified by C-18 reverse phase chromatography on a Luna column, using 10-50% ACN/water with 0.1% TFA as additive to give title compound (9.0 mg, 0.0181 mmol, 23.99 % yield). LCMS: m/z = 497.3 [M+H]+. Example 68: 2,2-difluoro-3-[(lR,3R)-3-methyl-l-[2-[(3-methylazetidin-l-yl)methyl]-lfI-indol- 5-yl]-l, 3, 4, 9-tetrahydropyrido[3, 4-b]indol-2-yl]propan-l -ol.
Figure imgf000133_0001
[0278] 3-Methylazetidin-l-ium; chloride was used to synthesize the title compound using the procedure described for Example 67 (7.0 mg, 0.0146 mmol, 19.36 % yield) LCMS: m/z = 479.2 [M+H]+.
Figure imgf000133_0002
[0279] To methyl 6-cyano-lH-indole-2-carboxylate (1.00 equiv, 0.13 g, 0.624 mmol) in DCM (10 mL) at 0 °C under nitrogen was added diisobutylaluminum hydride (4.00 equiv, 2.5 mL, 2.50 mmol) and stirred at 0 °C for 2 h. The reaction was quenched with ethyl acetate, followed by solution of Rochelle's salt, and stirred for 30 mins. The organic layer was separated and aqueous layer was extracted with additional DCM twice. The organic layer was dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-10% MeOH in DCM to give title compound (30 mg, 0.171 mmol, 27.43 % yield). 176.1 [M+H]+.
Example 70: [6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-2-yl]methanol.
Figure imgf000133_0003
[0280] To 2-fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l -amine (1.00 equiv, 43 mg, 0.171 mmol) in DCE (3 mL) was added 2-(hydroxymethyl)-lH-indole-6- carbaldehyde (1.00 equiv, 30 mg, 0.171 mmol) and acetic acid (2.00 equiv, 0.020 mL, 0.343 mmol) and stirred at 110 °C in microwave for 3 h. The reaction mixture was extracted with DCM, washed with saturated sodium bicarbonate solution, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using a 12 g column, eluting with 10-80% EtOAc in hexane, to give title compound (30 mg, 0.0740 mmol, 43.20 % yield). 406.2 [M+H]+.
Example 71: 6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde.
Figure imgf000134_0001
[0281] To [6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-2-yl]methanol (1.00 equiv, 30 mg, 0.0740 mmol) in DMSO (0.5 mL) was added the 1 -hydroxy- l,2-benxiodoxol-3(lH)-one 1-oxide (1.20 equiv, 55 mg, 0.089 mmol) and stirred at room temperature for 2 h. The reaction was quenched with water, extracted with ethyl acetate and layers separated. The EtOAc layer was washed with water, brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0- 40% EtOAc in hexane to give title compound (25 mg, 0.0620 mmol, 83.75 % yield). 404.2 [M+H]+.
Example 72: (lR,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-indol-6-yl]-2-(2-fluoro-
2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000134_0002
[0282] To a solution of 3-(fluoromethyl)azetidine hydrochloride (2.00 equiv, 16 mg, 0.124 mmol) in DMF (0.5 mL) was added triethylamine (2.00 equiv, 0.017 mL, 0.124 mmol) followed by 6-[(lR, 3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl- 1,3,4, 9-tetrahydropyrido[3,4-b]indol-l- yl]-lH-indole-2-carbaldehyde (1.00 equiv, 25 mg, 0.0620 mmol) in DMF (0.5 mL) and acetic acid (2.00 equiv, 0.0071 mL, 0.124 mmol). The reaction mixture was stirred for 1 h and treated with sodium triacetoxyborohydride (3.00 equiv, 39 mg, 0.186 mmol), and then stirred for additional 2 h. The reaction was quenched with 1 mL of methanol and purified by C-18 reverse phase chromatography using a Phenomenex Luna column, eluting with 10-50% ACN in water with 0.1% formic acid as modifier, to give title compound (4.5 mg, 0.00944 mmol, 15.24 % yield). 477.2 [M+H]+.
Example 73: Methyl 5-[(lR, 3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3, 4-b]indol-l -yl]benzofuran-2-carboxylate.
Figure imgf000135_0001
[0283] Methyl 5-formylbenzofuran-2-carboxylate (250 mg, 1.22 mmol) and 2-fluoro-N- [(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l-amine (320 mg, 1.29 mmol) were dissolved in DCE (10 mb) and acetic acid (280 pL, 4.9 mmol) was added. The solution was refluxed at 120 °C for 4 hours then cooled to ambient temperature. The solution was neutralized with saturated sodium bicarbonate solution and the organic layer was separated then washed with saturated brine solution. It was then dried over sodium sulfate, filtered, and dried in vacuo to yield the title compound (523 mg, 98% yield). LCMS: m/z = 435.2 [M+H]+. Example 74: [5-[(lR,3R)-2-(2-Eluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]benzofuran-2-yl]methanol.
Figure imgf000136_0001
[0284] Methyl 5-[(lR, 3R)-2-(2-fluoro-2-methyl-propyl)-3 -methyl- 1, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]benzofuran-2-carboxylate (110 mg, 0.25 mmol) was dissolved in dry THF under a nitrogen atmosphere and cooled to -78 °C. DIBAL-H (1.0 mmol in toluene) was added and stirred for 2 hours. The reaction was quenched with saturated ammonium chloride solution and diluted with EtOAc. The organic phase was separated and washed with saturated brine solution, dried over sodium sulfate and the volatiles were evaporated in vacuo. The crude was purified via flash chromatography in 0-40% EtOAc in hexanes to give the title product (58 mg, 56% yield). LCMS: m/z = 407.5 [M+H]+.
Example 75: 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]benzofuran-2-carbaldehyde.
Figure imgf000136_0002
[0285] [5-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]benzofuran-2-yl]methanol (58 mg, 0.14 mmol) and IBX (160 mg, 0.57 mmol) were dissolved in DMSO (1 mL) and stirred for 18 h. The reaction was quenched with saturated sodium bicarbonate solution and diluted with EtOAc. The organic layer was separated and washed with saturated brine solution, then dried over sodium sulfate. The volatiles were removed under reduced pressure. The crude was purified via flash chromatography in 0-40% EtOAc in hexanes to give the title compound (37 mg, 64% yield). LCMS: m/z = 405.3 [M+H]+. Example 76: ( 1 R,3R)~ I -[2-[[3-(fluoromethyl)azetidin- 1 -yl]methyl]benzofuran-5-yl]-2-(2- fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000137_0001
[0286] 5-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]benzofuran-2-carbaldehyde (0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 3-(fluoromethyl)azetidine hydrochloride (0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. Sodium triacetoxyborohydride (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kinetix 5 pm Cl 8 100 A column (size: 100 x 30.0 mm; gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white, fluffy, solid (7.3 mg, 31% yield). LCMS: m/z = 478.3 [M+H]+.
Example 77: (lR,3R)-l-[2-[(6,6-difluoro-l-azaspiro[3.3]heptan-l-yl)methyl]benzofuran-5-yl]-
2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000137_0002
[0287] 5-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]benzofuran-2-carbaldehyde (17 mg, 0.042 mmol) was dissolved in DMF (0.5 mL) and acetic acid (5 pL, 0.084 mmol) was added. Separately, bis(6,6-difluoro-l- azaspiro[3.3]heptane); oxalic acid (7.5 mg, 0.025 mmol) was dissolved in DMF (0.5 mL) and TEA (12 pL, 0.084 mmol) was added. These solutions were combined and stirred for 30 minutes. Sodium triacetoxyborohydride (27 mg, 0.12 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kinetix 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white, fluffy, solid (8.5 mg, 39% yield). LCMS: m/z = 522.3 [M+H]+.
Example 78: Methyl 3-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-5-carboxylate.
Figure imgf000138_0001
[0288] To a microwave vial equipped with a magnetic stir bar was added a solution of 2- fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l-amine (200 mg, 0.805 mmol) in anhydrous DCE (4 mL). To the solution was added methyl 3-formyl-lH-pyrrolo[2,3- b]pyridine-5-carboxylate (160 mg, 0.784 mmol) and acetic acid (0.20 mL, 3.49 mmol). The reaction vessel was closed and purged with nitrogen. The reaction was heated in microwave reactor at 110 °C for 1 h. Saturated solution of sodium bicarbonate (20 mL) was added to the reaction mixture and the aqueous layer was extracted with EtOAc (15 mL x 3). The combined organic layer was washed with brine (10 mL), dried over anhydrous Na2SO4, fdtered and then concentrated under reduced pressure to give crude product. The crude was purified by Combiflash column chromatography (silica gel 12 g, EtOAc:Hex 0-100 %) to give 256 g (73 %) of titled compound. LCMS: m/z =435.4 [M+H]+.
Example 79: [3-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lE[-indazol-5-yl]methanol
Figure imgf000138_0002
[0289] To a reaction vial equipped with a magnetic stir bar and a nitrogen inlet was added a solution of methyl 3-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indazole-5-carboxylate (236 mg, 0.543 mmol) in anhydrous THF (5 mL). To the solution was added 1 N lithium aluminum hydride in THF (1.5 mb, 1.50 mmol) dropwise at 0 °C. The reaction mixture was then warmed to room temperature and stirred for 30 minutes. Upon completion, the reaction was cooled to 0 °C. To the solution was added, 0.10 mL H2O was dropwise, then 0.20 mL 10% NaOH solution and lastly followed by addition of 0.40 mL H2O. The reaction mixture was stirred at rt for 30 min, fdtered and then concentrated under vacuum to afford crude product as pale-yellow solid. The crude was purified by Combiflash column chromatography (DCM MeOH 0-20% and 2% TEA as modifier) to give the titled product (171 mg, 0.421 mmol, 77 % yield) as pale-yellow oil. LCMS: m/z =407.2 [M+H]+.
Example 80: 3-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l-yl]-lH-indazole-5-carbaldehyde.
Figure imgf000139_0001
[0290] To a solution of [3-[(lS,3R)-2-(2-fhioro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indazol-5-yl]methanol (164 mg, 0.403 mmol) in anhydrous DMSO (2 mL) was added 2-iodoxybenzoic acid (250 mg, 0.402 mmol). The reaction mixture was stirred for 1 hour. Upon completion, water (20 mL) was added to the reaction mixture and the aqueous layer was extracted with DCM (15 mL x 3). The combined organic layer was dried over anhydrous Na2SO4, filtered and then concentrated. The crude was purified by Teledyne Isco System (silica gel 12 g, MeOH:DCM 0-20 %) to give title compound (110 mg, 0.272 mmol, 67.41 % yield) as pure and desired product. LCMS: m/z =405.2 [M+H]+. Example 81: (1 R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl- 1 -[5-[(3-methylazetidin- 1 - yl)methyl]-lH-pyrrolo[2,3-b]pyridin-3-yl]-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000140_0001
[0291] To a solution of 3-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-5-carbaldehyde (34 mg, 0.0841 mmol) in DMF (0.50 mb) was added a solution of 3 -methylazetidine hydrochloride (18 mg, 0.167 mmol) in DMF (0.50 mL) and triethylamine (0.023 mL, 0.165 mmol). To the reaction mixture was added acetic acid (0.019 mL, 0.332 mmol) and stirred at room temperature for 1 h and then treated with sodium triacetoxyborohydride (53 mg, 0.250 mmol) and triethylamine (0.023 mL, 0.165 mmol) and stirred for 1 h. The reaction was quenched with 1 mL of methanol and the crude directly purified by C-l 8 reverse phase chromatography using prep HPLC, eluting with 10-50% ACN in water with 0.1% formic acid as modifier to give title compound (20 mg, 0.0418 mmol, 49.70 % yield) as yellow solid. LCMS: m/z =460.3 [M+H]+.
Example 82: (lR,3R)-l-[5-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-pyrrolo[2,3-b]pyridin-3- yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000140_0002
[0292] To a solution of 3-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-5-carbaldehyde (34 mg, 0.0841 mmol) in DMF (0.5 mL) was added a solution of 3-(fluoromethyl)azetidine hydrochloride (21 mg, 0.167 mmol) in DMF (0.50 mL) and triethylamine (0.023 mL, 0.165 mmol). To the reaction mixture was added acetic acid (0.019 mL, 0.332 mmol) and stirred at room temperature for 1 h. The reaction mixture was then treated with sodium triacetoxyborohydride (53 mg, 0.250 mmol) and triethylamine (0.023 mL, 0.165 mmol) and stirred for 1 h. The reaction was quenched with 1 mL of methanol and the crude directly purified by C-18 reverse phase chromatography using prep HPLC, eluting with 10-50% ACN in water with 0.1% formic acid as modifier to give title compound (16 mg, 0.0322 mmol, 38.26 % yield) as yellow solid. LCMS: m/z =478.4 [M+H]+.
Example 83: (lR,3R)-l-[5-(3-azabicyclo[3.1.0]hexan-3-ylmethyl)-lEI-pyrrolo[2,3-b]pyridin-3- yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000141_0001
[0293] To a solution of 3-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-5-carbaldehyde (33 mg, 0.0816 mmol) in DMF (0.5 mb) was added a solution of 3-azabicyclo[3.1.0]hexane hydrochloride (20 mg, 0.167 mmol) in DMF (0.5 mL) and triethylamine (0.023 mL, 0.165 mmol). To the reaction mixture was added acetic acid (0.019 mL, 0.332 mmol) and stirred at room temperature for 1 h. The reaction mixture was then treated with sodium tri acetoxyb orohydri de (52 mg, 0.245 mmol) and triethylamine (0.023 mL, 0.165 mmol) and stirred for 1 h. The reaction was quenched with 1 mL of methanol and the crude directly purified by C-18 reverse phase chromatography using prep HPLC, eluting with 10-50% ACN in water with 0.1% formic acid as modifier, to give title compound (20 mg, 0.0394 mmol, 48.34 % yield) as yellow solid. LCMS: m/z =473.0 [M+H]+.
Example 84: Methyl 2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-6H-thieno[2,3-b]pyrrole-5-carboxylate.
Figure imgf000141_0002
[0294] 2-Fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l -amine (550 mg, 2.2 mmol) was dissolved in DCE (13 mL). Methyl 2-formyl-6H-thieno[2,3-b]pyrrole-5- carboxylate (500 mg, 2.4 mmol) and acetic acid (1.27 mL, 22.1 mmol) were added to the solution and microwaved at 150 °C for 30 min. The volatiles were removed under reduced pressure and the crude was purified by flash chromatography on silica with 0-50% EtOAc in hexanes to yield the title compound (174 mg, 18% yield). LCMS: m/z = 440.9 [M+H]+.
Example 85: [2-[(lS,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-6H-thieno[2,3-b]pyrrol-5-yl]methanoL
Figure imgf000142_0001
[0295] Methyl 2-[(l S, 3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-6H-thieno[2,3-b]pyrrole-5-carboxylate (160 mg, 0.36 mmol) was dissolved under an atmosphere of nitrogen in anhydrous THF and cooled to 0 °C. 1 M LAH (1.2 mmol) in THF was added and stirred for 3 hours. The reaction was quenched with EtOAc, then washed with saturated Rochelle's salt solution, then water. The combined organic layers were dried over sodium sulfate, filtered, then evaporated under reduced pressure. The crude was purified by flash chromatography on 12 g silica column with 0-50% EtOAc in hexanes to give the title compound (35 mg, 23% yield). LCMS: m/z = 412.2 [M+H]+.
Example 86: 2-[(lS,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-6H-thieno[2,3-b]pyrrole-5-carbaldehyde.
Figure imgf000143_0001
[0296] [2-[(lS,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-6H-thieno[2,3-b]pyrrol-5-yl]methanol (35 mg, 0.085 mmol) and IBX (23.8 mg, 0.085 mmol) were dissolved in DMSO and stirred for 1.5 hours. The reaction was quenched with water, then extracted with EtOAc. The combined organic layers were washed with saturated brine solution and water, dried over sodium sulfate, filtered, and evaporated under reduced pressure. The crude was purified by flash chromatography on a TEA-rinsed 12 g silica column with 0-30% EtOAc in hexanes to yield the title compound (22 mg, 63% yield). LCMS: m/z = 410.2 [M+H]+.
Example 87: 2-[(lS,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- bJindol-l-ylJ-5-[(3-methylazetidin-l-yl)methylJ-6H-thieno[2,3-bJpyrrole.
Figure imgf000143_0002
[0297] To solution of 2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-6H-thieno[2,3-b]pyrrole-5-carbaldehyde (9.0 mg, 0.022 mmol) in DMF (0.5 mL) was added acetic acid (2.6 mg, 0.044 mmol). Separately, 3- methylazetidin-l-ium chloride (2.4 mg, 0.022 mmol) was dissolved in DMF (0.5 mL) and TEA (4.4 mg, 0.044 mmol) was added. These solutions were combined and stirred for 30 min. Sodium triacetoxyborohydride (14.0 mg, 0.066 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL of MeOH and directly purified by HPLC on Kinetix 5 pm C 18 100A column (size 100 x 30.0 mm; gradient 5-50% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to obtain the title compound as an off-white powder (2.2 mg, 21% yield). LCMS: m/z = 465.2 [M+H]+.
Example 88: 5-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-2-[(lS,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-6H-thieno[2,3-b]pyrrole.
Figure imgf000144_0001
[0298] To solution of 2-[(l S,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l , 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-6H-thieno[2,3-b]pyrrole-5-carbaldehyde (12.6 mg, 0.030 mmol) in DMF (0.5 mL) was added acetic acid (3.7 mg, 0.062 mmol). Separately, 3- (fluoromethyl)azetidin-l-ium chloride (3.9 mg, 0.031 mmol) was dissolved in DMF (0.5 mL) and TEA (6.2 mg, 0.062 mmol) was added. These solutions were combined and stirred for 30 min. Sodium triacetoxyborohydride (20.0 mg, 0.092 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL of MeOH and directly purified by HPLC on Kinetix 5 pm C18 100A column (size 100 x 30.0 mm; gradient 5-50% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to obtain the title compound as a pink powder (2.7 mg, 18% yield). LCMS: m/z = 483.2 [M+H]+.
Example 89: Methyl 2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole-5-carboxylate.
Figure imgf000144_0002
[0299] To 2-fluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]-2-methyl-propan-l -amine (1.00 equiv, 300 mg, 1.21 mmol) in DCE (20 mL) was added the methyl 2-formyl-4H- thieno[3,2-b]pyrrole-5-carboxylate (1.20 equiv, 303 mg, 1.45 mmol), followed by acetic acid (2.50 equiv, 0.17 mL, 3.02 mmol) in a microwave tube, flushed with nitrogen. The reaction tube was capped and heated at 100 °C in the microwave for 3 h. The reaction was cooled to room temperature and extracted with DCM. The organic layer was washed with saturated sodium bicarbonate, dried over sodium sulfate, filtered and concentrated. The crude was purified by silica gel chromatography using 0-10% EtOAc in DCM to give the title compound (380 mg, 0.865 mmol, 71% yield). LCMS: m/z = 440.2 [M+H]+.
Example 90: 2-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l -yl]-4H-thieno[3,2-b]pyrrole-5-carboxylic acid.
Figure imgf000145_0001
[0300] To methyl 2-[(lS,3R)-2-(2-fhioro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole-5-carboxylate (1.00 equiv, 23 mg, 0.0523 mmol) in THF (3 mL)/methanol (1 mL)/water (0.5 mL) was added the lithium hydroxide (16.0 equiv, 20 mg, 0.835 mmol) and stirred at 60 °C for 16 h. The reaction mixture was treated with formic acid (16.0 equiv, 0.032 mL, 0.837 mmol), and concentrated under reduced pressure. The crude was taken up in methanol and purified by reverse phase chromatography using Luna C-18 column eluting with water/ACN with 0.1% formic acid as additive to give the title compound (8.0 mg, 0.0181 mmol, 34.5 % yield). LCMS: m/z = 426.0 [M+H]+. Example 91: [2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-4H-thieno[3,2-b]pyrrol-5-yl]methanol.
Figure imgf000146_0001
[0301] To methyl 2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole-5-carboxylate (1.00 equiv, 200 mg, 0.455 mmol) in THF (5 mL) under an atmosphere of nitrogen at 0 °C was added the lithium aluminum hydride (3.00 equiv, 1.4 mL, 1.37 mmol) dropwise. The reaction mixture was allowed to warm up to room temperature and stirred for 2 h. The reaction mixture was cooled in an ice water bath and diluted with MTBE and few drops of water, then a solution of sodium hydroxide (0.659 equiv, 0.080 mL, 0.300 mmol) was added. The reaction mixture was stirred vigorously for 15 mins and then filtered through Celite®. The filtrate was concentrated, the residue taken up in ethyl acetate, the EtOAc layer was washed with brine, dried over sodium sulfate, filtered and concentrated. The crude was purified by silica gel chromatography using 10-80% EtOAc in DCM to give the title compound (140 mg, 0.340 mmol, 74% yield). LCMS: m/z = 413.3 [M+H]+.
Example 92: 2-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l-yl]-4H-thieno[3,2-b]pyrrole-5-carbaldehyde.
Figure imgf000146_0002
[0302] To [2-[(l S,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-4H-thieno[3,2-b]pyrrol-5-yl]methanol (1.00 equiv, 50 mg, 0.121 mmol) in DCM was added manganese dioxide (20.0 equiv, 211 mg, 2.43 mmol) and stirred at room temperature for 48 h. The reaction mixture was filtered through Celite®, and concentrated. The crude was purified by silica gel chromatography using 0-50% EtOAc in DCM to give the title compound (20 mg, 0.0488 mmol, 40.2% yield). LCMS: m/z = 410.2 [M+H]+.
Example 93: 5-[[3-(fluoromethyl)azetidin-l-yl]methyl]-2-[(lS,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole.
Figure imgf000147_0001
[0303] To 2-[(l S,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-4H-thieno[3,2-b]pyrrole-5-carbaldehyde (1.00 equiv, 20 mg, 0.0488 mmol) in DMF (1 mL) and acetic acid (2.00 equiv, 0.0056 mL, 0.0977 mmol) was added 3- (fluoromethyl)azetidin-l-ium; chloride (2.0 equiv, 12 mg, 0.0977 mmol) dissolved in DMF (0.5 mL) and triethylamine (2.00 equiv, 0.014 mL, 0.0977 mmol) under nitrogen. The reaction mixture was stirred at room temperature for 1 h and to this was added the sodium triacetoxyborohydride (3.00 equiv, 31 mg, 0.147 mmol). The reaction mixture was stirred at room temp for 2 h, then quenched with methanol. The crude was purified by C-18 reverse phase chromatography eluting with ACN/Water with 0.1 % formic acid as modifier to give the title compound (4.3 mg, 0.00891 mmol, 18.2 % yield). LCMS: m/z = 483.0 [M+H]+.
Example 94: 2-[( IS, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl- 1, 3,4, 9-tetrahydropyrido[3, 4- b]indol-l-yl]-5-[(3-methylazetidin-l-yl)methyl]-4H-thieno[3,2-b]pyrrole.
Figure imgf000148_0001
[0304] Prepared by the method of Example 93 using 3-methylazetidin-l-ium; chloride LCMS: m/z = 465.0 [M+H]+.
Example 95: 3-fluoro-N-[[2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrol-5-yl]methyl]propan-l-amine.
Figure imgf000148_0002
[0305] Prepared by the method of Example 93 using 3 -fluoropropyl ammonium; chloride.
LCMS: m/z = 471.0 [M+H]+.
Example 96: 5-[[(3S)-3-ethylpyrrolidin-l-yl]methyl]-2-[(lS,3R)-2-(2-fluoro-2-methyl-propyl)- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-4H-thieno[3,2-b]pyrrole.
Figure imgf000149_0001
[0306] Prepared by the method of Example 93 using (3S)-3-ethylpyrrolidin-l-ium;chloride. LCMS: m/z = 493.0 [M+H]+.
Example 97: Methyl 5-bromo-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-2- carboxylate.
Figure imgf000149_0002
[0307] Methyl 5-bromo-lH-pyrrolo[2,3-b]pyridine-2-carboxylate (944 mg, 3.7 mmol) was dissolved in DMF (10 mb) and brought to 0 °C. Sodium hydride (98 mg, 4.1 mmol) was added and stirred for 30 min. 2-(Trimethylsilyl)ethoxymethyl chloride (679 mg, 4.1 mmol) was added and stirred for 2 h. The reaction was quenched with saturated sodium bicarbonate solution and diluted with EtOAc. The organic layer was separated and washed with saturated brine solution 5 times. It was then dried over sodium sulfate, filtered, and concentrated in vacuo to give the title product (1.277 g, 90% yield). LCMS: m/z = 386.2 [M+H]+. Example 98: Methyl 5-cyano-l-(2-trimethylsilyletlioxymetliyl)pyrrolo[2,3-b]pyridine-2- carboxylate.
Figure imgf000150_0001
[0308] Methyl 5-bromo-l -(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-2- carboxylate (1.27 g, 3.3 mmol), zinc cyanide (464 mg, 4.0 mmol), [1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium(II) (241 mg, 0.33 mmol), tris(dibenzylideneacetone)dipalladium(0) (151 mg, 0.16 mmol), and zinc (54 mg, 0.82 mmol) were dissolved in DMF (15 mL) under an atmosphere of nitrogen. The reaction was heated to 120 °C and stirred for 16 h. The reaction was brought to room temperature then filtered. It was diluted with EtOAc then partitioned with saturated sodium bicarbonate solution.
The organic layer was washed with saturated brine solution 5 times, dried over sodium sulfate, filtered, then dried in vacuo. The crude material was purified on silica gel in 0-50% EtOAc in hexanes gradient to yield the title product (711 mg, 65% yield). LCMS: m/z = 332.5 [M+H]+.
Example 99: 2-(Hydroxymethyl)-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-bJpyridine-5- carbaldehyde.
Figure imgf000150_0002
[0309] Methyl 5-cyano-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-2- carboxylate (711 mg, 2.1 mmol) was dissolved in THF (7 mL) in an atmosphere of nitrogen and brought to 0 °C. DIBAL (6.4 mmol) was added dropwise and stirred for 90 min. The reaction was brought to room temperature and quenched with EtOAc. Saturated Rochelle's salt solution was added and stirred for 45 min. The organic layer was then separated and washed with saturated brine solution, dried over sodium sulfate, filtered, and dried in vacuo. The crude material was purified on silica gel in 0-10% methanol in DCM gradient to give the title product (323 mg, 49% yield). LCMS: m/z = 307.4 [M+H]+.
Example 100: [5-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-2- yljmethanol.
Figure imgf000151_0001
[0310] 2-(Hydroxymethyl)-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-5- carbaldehyde (160 mg, 0.52 mmol), (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2- methylpropan-1 -amine (130 mg, 0.52 mmol) were dissolved in DCE (2 mL) and acetic acid (1.5 mL, 2.6 mmol) was added. The solution was stirred at 120 °C with microwave irradiation for 2 h. The solution was poured into saturated sodium bicarbonate solution and the organic layer was separated and dried over sodium sulfate. It was filtered then dried in vacuo to give the title compound (280 mg, 100%). LCMS: m/z = 537.8 [M+H]+.
Example 101: 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-2-carbaldehyde.
Figure imgf000151_0002
[0311] [5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridin-2-yl]methanol (280 mg, 0.52 mmol) and IBX (219 mg, 0.78 mmol) were dissolved and stirred in DMSO (2 mL) for 1.5 h. The reaction was quenched with saturated sodium bicarbonate solution and diluted with EtOAc. The organic phase was separated and washed with saturated brine solution. It was then dried over sodium sulfate, fdtered, and dried in vacuo. The crude was purified on silica gel in 0-40% EtOAc in hexanes gradient to yield the title compound (101 mg, 36% yield). LCMS: m/z = 535.7 [M+H]+.
Example 102: 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-2-carbaldehyde.
Figure imgf000152_0001
[0312] 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(2-trimethylsilylethoxymethyl)pyrrolo[2,3-b]pyridine-2-carbaldehyde (101 mg, 0.19 mmol) was dissolved in THF (2 mL) and TBAF (1.9 mmol) was added. The reaction was heated to 80 °C and stirred for 16 h. The reaction was quenched with saturated sodium bicarbonate solution and diluted with EtOAc. The organic phase was separated and washed with saturated brine solution, dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH in DCM gradient to yield the title compound (70 mg, 92% yield). LCMS: m/z = 405.4 [M+H]+.
Example 103: (lR,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-pyrrolo[2,3-b]pyridin- 5-yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000152_0002
[0313] 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-pyrrolo[2,3-b]pyridine-2-carbaldehyde (26 mg, 0.06 mmol) was dissolved in
DMF (1 mL) and acetic acid (7.5 pL, 0.13 mmol) was added. Separately, 3- (fluoromethyl)azetidine hydrochloride (10 mg, 0.08 mmol) was dissolved in DMF (1 mL) and TEA (18 pL, 0.13 mmol) was added. These solutions were combined and heated at 60 °C for 30 min. STAB (41 mg, 0.19 mmol) was added and stirred for 2 h. The reaction was quenched with MeOH (1 mL) and directly purified by HPLC on a Kintetex 5 pm Cl 8 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (13 mg, 42% yield). LCMS: m/z = 478.3 [M+H]+.
Example 104: Ethyl 5-chloro-l-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridine-2- carboxylate.
Figure imgf000153_0001
10314] To ethyl 5-chloro-lH-pyrrolo[3,2-b]pyridine-2-carboxylate (1.00 equiv, 500 mg, 2.23 mmol) in anhydrous DMF (5 mL) under nitrogen at 0 °C was added the sodium hydride (1.20 equiv, 107 mg, 2.67 mmol) and stirred for 30 min. To this was added 2- (tri methyl silyl )ethoxymethyl chloride (1.20 equiv, 0.47 mL, 2.67 mmol) and stirred for 1 h. The reaction was quenched with water, extracted with EtOAc and the layers separated. The EtOAc layer was washed with water, brine, dried over sodium sulfate, filtered and concentrated. The crude was purified by silica gel chromatography using a 24 g column, eluting with 0-15% EtOAc in hexane to give title compound (730 mg, 2.06 mmol, 92.4 % yield). LCMS: m/z = 355.1 [M+H]+.
Example 105: Ethyl 5-cyano-l-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridine-2- carboxylate
Figure imgf000154_0001
[0315] To ethyl 5-chloro-l-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridine-2- carboxylate (1.00 equiv, 710 mg, 2.00 mmol) in NMP (10 mL) was added the zinc cyanide (6.00 equiv, 1409 mg, 12.0 mmol), and the reaction mixture sparged with nitrogen. To this was added the tetrakis(triphenylphosphine)palladium(0) (0.0500 equiv, 116 mg, 0.100 mmol), and the mixture sparged with nitrogen for an additional 5 min. The reaction was capped and heated at 120 °C for 18 h. The reaction was filtered through Celite®, and the residue was washed with EtOAc (50 mL). The filtrate was washed with water, brine, dried over sodium sulfate, filtered and concentrated. The crude was purified by silica gel chromatography using 24 g column, eluting with 0-20% EtOAc in hexane to give title compound (375 mg, 1.09 mmol, 54.3 % yield). LCMS: m/z = 346.2 [M+H]+.
Example 106: Ethyl 5-formyl-l-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-h]pyridine-2- carboxylate
Figure imgf000154_0002
[0316] To ethyl 5-cyano-l-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridine-2- carboxylate (1.00 equiv, 410 mg, 1.19 mmol) in THF (lOmL) at 0 °C under nitrogen was added the diisobutylaluminum hydride (1.50 equiv, 1.8 mL, 1.78 mmol) dropwise and stirred for 1 h after slowly warming to room temp. The reaction was cooled back to 0 °C, quenched with
EtOAc, followed by saturated solution of Rochelle's salt, and stirred for 30 min. The reaction mixture was extracted with EtOAc, layers were separated, and the organic layer dried over sodium sulfate. The EtOAc layer was filtered, concentrated under vacuum, and purified by silica gel chromatography using 0-30% EtOAc in hexane to give ethyl 5-formyl-l-(2- trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridine-2-carboxylate (210 mg, 0.603 mmol, 50.8 % yield). LCMS: m/z = 349.1 [M+H]+.
Example 107: Ethyl 5-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-b]pyridine-2- carboxylate
Figure imgf000155_0001
[0317] To (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (1.20 equiv, 191 mg, 0.768 mmol) in DCE (3 mL) was added ethyl 5-formyl-l-(2- trimethylsilylethoxymethyl)pyrrolo[2,3-c]pyridine-2-carboxylate (1.00 equiv, 223 mg, 0.640 mmol) and acetic acid (2.00 equiv, 0.073 mL, 1.28 mmol), and the mixture was stirred at 110 °C in microwave for 3 h. The reaction mixture was extracted with DCM, washed with saturated sodium bicarbonate solution, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using a 24 g column elution with 0-30% EtOAc in hexane to give title compound (256 mg, 0.442 mmol, 69.1 % yield). LCMS: m/z = 579.8 [M+H]+.
Example 108: (5-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lEI- pyrido[3,4-b]indol-l-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-b]pyridin-2- yl) methanol
Figure imgf000156_0001
[0318] To ethyl 5-((lS,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-17/- pyrido[3,4-Z>]indol-l-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-l/7-pyrrolo[3,2-/>]pyridine-2- carboxylate (1.00 equiv, 256 mg, 0.442 mmol) in THF (5mL) at 0 °C under atmosphere of nitrogen was added lithium aluminum hydride (2.00 equiv, 0.88 mL, 0.885 mmol) and stirred for 30 min. The reaction was quenched with EtOAc and saturated solution of Rochelle's salt, and stirred for 30 min. The reaction mixture was extracted with EtOAc, layers separated, and the organic layer dried over sodium sulfate. The EtOAc layer was filtered, concentrated under vacuum, and purified by silica gel chromatography using 0-30% EtOAc in DCM to give title compound (210 mg, 0.391 mmol, 88.5% yield). LCMS: m/z = 537.8 [M+H]+.
Example 109: (5-((lS,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-
Figure imgf000156_0002
[0319] A mixture of (5-((15,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- lJ77-pyrido[3,4-Z>]indol-l-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-l//-pyrrolo[3,2-£>]pyri din-2- yl)methanol (1.00 equiv, 120 mg, 0.224 mmol) in DCM (2 mL) was cooled to 0 °C and treated with trifluoroacetic acid (58.1 equiv, 1.0 mL, 13.0 mmol). The reaction mixture was stirred for 1 h. The reaction mixture was concentrated under vacuum, extracted with EtOAc, neutralized with sat. NaHCCh, dried over sodium sulfate, filtered, and concentrated. The crude was dissolved in methanol (1 mL) and treated with triethylamine (4.49 equiv, 0.14 mL, 1.00 mmol) and heated at 90 °C for 1 h, concentrated under vacuum, extracted with EtOAc, washed with water, brine, dried over sodium sulfate, filtered, and concentrated to give title compound (60 mg, 0.148 mmol, 66.0 % yield). LCMS: m/z = 407.2 [M+H]+.
Example 110: 5-[(l S, 3R)-2- (2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4, 9-tetrahydropyrido[3, 4-
Figure imgf000157_0001
[0320] To [5-[(l S,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l ,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-pyrrolo[3,2-b]pyridin-2-yl]methanol (1.00 equiv, 60 mg, 0.148 mmol) in anhydrous DMSO (1 mL) was added l,l,l-tris(acetyloxy)-l,l-dihydro-l,2-benzodioxol-3-(lh)- one (1.50 equiv, 209 mg, 0.221 mmol) and stirred at RT for 1 h. The reaction was quenched with water and extracted with EtOAc. The EtOAc layer was washed with IN NaOH and brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-30% EtOAc in DCM to give title compound (45 mg, 0.111 mmol, 75.4 % yield). LCMS: m/z = 405.3 [M+H]+.
Example 111: (lS,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-lH-pyrrolo[3,2-b]pyridin-
Figure imgf000157_0002
[0321] To 3-(fluoromethyl)azetidine hydrochloride (2.00 equiv, 28 mg, 0.223 mmol) in NMP (1 mL) was added the triethylamine (2.00 equiv, 0.031 mL, 0.223 mmol), stirred for 5 min, followed by addition of 5-[(lS,3R)-2-(2-fhioro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-pyrrolo[3,2-b]pyridine-2-carbaldehyde (1.00 equiv, 45 mg, 0.111 mmol) and acetic acid (2.00 equiv, 0.013 mL, 0.223 mmol). The reaction mixture was stirred at room temp for 30 min, followed by addition of sodium triacetoxyborohydride (3.00 equiv, 71 mg, 0.334 mmol) and stirred for 1 h. The reaction was quenched with methanol (0.5 mL) and purified by C-18 reverse phase chromatography using Luna column, eluting with 5-30% ACN in water with 0.1% formic acid as modifier to give title compound (29 mg, 0.0607 mmol, 54.6 % yield). LCMS: m/z = 478.3 [M+H]+.
[0322] XH NMR (400 MHz, de-DMSO) 5 11.15 (d, J = 2.0 Hz, 1H), 10.53 (s, 1H), 8.15 (s, 1H), 7.63 (dd, J = 8.4, 0.9 Hz, 1H), 7.41 (d, J = 7.7 Hz, 1H), 7.28 - 7.20 (m, 2H), 6.98 (dtd, J = 23.6, 7.0, 1.2 Hz, 2H), 6.32 (d, J = 1.8 Hz, 1H), 5.10 (s, 1H), 4.58 (d, J = 6.1 Hz, 1H), 4.46 (d, J = 6.1 Hz, 1H), 3.69 (s, 2H), 3.01 (t, J = 6.6 Hz, 2H), 2.91 - 2.63 (m, 3H), 1.38 (d, J = 21.7 Hz, 3H), 1.28 (d, J = 21.6 Hz, 3H), 1.09 (d, J = 6.7 Hz, 3H).
Example 112: 2-[(3-methylazetidin-l-yl)methylJ-3l l-benzimidazole-5-carbonitrile.
Figure imgf000158_0001
[0323] 2-(Chloromethyl)-3H-benzimidazole-5-carbonitrile (161 mg, 0.84 mmol) and 3- methylazetidine hydrochloride (135 mg, 1.2 mmol) were dissolved in DCM (3 mL) and TEA (3.4 mmol) was added. The solution was stirred for 4 h at 40 °C. The solution was cooled then washed with brine solution. The organic layer dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-15% MeOH in DCM to give the titled product (60 mg, 32% yield). LCMS: m/z = 227.2 [M+H]+. Example 113: 2-[(3-methylazetidin-l-yl)methyl]-3H-benzimidazole-5-carbaldeliyde.
Figure imgf000159_0002
[0324] 2-[(3-Methylazetidin-l-yl)methyl]-3H-benzimidazole-5-carbonitrile (60 mg, 0.27 mmol) was dissolved in THF (2 mL) under nitrogen at 0 °C. DIBAL (0.53 mmol) was added and stirred for 1 h. The reaction was quenched with EtOAc. Rochelle's Salt solution was added and stirred for 1 h. The organic layer was separated and washed with brine solution, dried over sodium sulfate, filtered, and evaporated in vacuo to yield the title compound (40 mg, 66% yield). LCMS: m/z = 230.3 [M+H]+.
Example 114: (lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l-[2-[(3-methylazetidin-l- yl) methyl]-3H-benzimidazol-5-yl]-l,3, 4, 9-tetrahydropyrido[3 , 4-b/indole.
Figure imgf000159_0001
[0325] 2-[(3-methylazetidin-l-yl)methyl]-3H-benzimidazole-5-carbaldehyde (40 mg, 0.17 mmol) and (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (43 mg, 0.17 mmol) were dissolved in DCE (2 mL) and acetic acid (50 pL, 0.87 mmol) was added. The solution was stirred at 120 °C under microwave irradiation for 2 h. The reaction was quenched with saturated sodium bicarbonate solution and the organic layer was separated, dried over sodium sulfate, filtered, and dried in vacuo. The crude was dissolved in DMF (2 mL) and purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5- 30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (13 mg, 42% yield). LCMS: m/z = 460.3 [M+H]+. Example 115: Methyl 2-[[3-(fluoromethyl)azetidin-l-yl]metliyl]-l,3-benzoxazole-6- carboxylate.
Figure imgf000160_0001
[0326] Methyl 2-(chloromethyl)-l,3-benzoxazole-6-carboxylate (250 mg, 1.1 mmol) and 3- (fluoromethyl)azetidine hydrochloride (167 mg, 1.3 mmol) were dissolved in DCM (4 mL) and triethylamine (620 pL, 4.4 mmol) was added. The solution was heated to 40 °C and stirred for 3 h. It was cooled to room temperature then poured into saturated sodium bicarbonate solution. The organic layer was separated, dried over sodium sulfate, fdtered, and dried in vacuo to give the title compound (320 mg, 104% yield). LCMS: m/z = 279.2 [M+H]+. This crude was used in the next step without further purification.
Example 116: [2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazol-6-yl]methanol.
Figure imgf000160_0002
[0327] Methyl 2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-6-carboxylate (320 mg, 1.1 mmol) was dissolved in THF (5 mL) under nitrogen and cooled to -78 °C. LAH (1.5 mmol) was added and stirred for 1 h. The solution was warmed to 0 °C and diluted with MTBE (10 mL). While stirring, 240 pL of water was added, then 240 pL of IN NaOH. The mixture was warmed to room temperature and stirred for 15 min. Sodium sulfate was added and stirred for another 15 min. This was then filtered and dried in vacuo to give the title compound (313 mg, 109% yield). LCMS: m/z = 251.4 [M+H]+. The crude was used in the next step without further purification. Example 117: 2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-6-carbaldeliyde.
Figure imgf000161_0001
[0328] [2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazol-6-yl]methanol (280 mg, 1.1 mmol) and IBX (313 mg, 1.1 mmol) were dissolved in DMSO (4 mL) and stirred for 2 h. The solution was diluted with EtOAc, then poured into saturated sodium bicarbonate solution, then the organic layer was separated and extracted with EtOAc 3 times. The combined organic layers were washed with saturated brine solution 3 times, dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH in DCM to give the title compound (33 mg, 12% yield). LCMS: m/z = 249.1 [M+H]+.
Example 118: 2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-6-[(lR,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l, 3, 4, 9-tetrahydropyrido[3, 4-b]indol-l -yl]-l, 3-benzoxazole.
Figure imgf000161_0002
[0329] (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (33 mg, 0.13 mmol) and 2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-6-carbaldehyde (33 mg, 0.13 mmol) were dissolved in DCE (1.5 mL) and acetic acid (38 pL, 0.66 mmol) was added. The solution was stirred at 130 °C under microwave irradiation for 2.5 h. The solution was then cooled to room temperature, diluted with DCM, and poured into saturated sodium bicarbonate solution. The organic layers were separated, dried over sodium sulfate, filtered, and dried in vacuo. The crude material was dissolved in DMF (2 mL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0,1% formic acid in water) then lyophilized to give the title compound as an off-white solid (3.1 mg, 5% yield). LCMS: m/z = 479.6 [M+H]+. 'H NMR (400 MHz, DMSO) 5 10.80 (s, 1H), 8.34 (s, 2H), 7.42 (d, J = 7.7 Hz, 1H), 7.27 (d, J = 8.0 Hz, 1H), 7.04 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.97 (td, J = 7.4, 1.1 Hz, 1H), 6.85 (d, J = 8.0 Hz, 1H), 6.74 (dd, J = 8.0, 1.9 Hz, 1H), 6.67 (d, J = 1.8 Hz, 1H), 4.99 - 4.93 (m, 1H), 4.66 (d, J = 5.7 Hz, 1H), 4.54 (d, J = 5.7 Hz, 1H), 4.46 (d, J = 2.2 Hz, 2H), 4.14 (s, 2H), 3.86 (s, 2H), 3.19 - 2.98 (m, 2H), 2.72 (dd, J = 31.2, 15.0 Hz, 1H), 1.45 (d, J = 21.8 Hz, 3H), 1.29 (d, J = 21.5 Hz, 3H), 1.06 (d, J = 6.7 Hz, 3H).
Example 119: Ethyl 6-bromoimidazo[l,2-a]pyridine-2-carboxylate.
Figure imgf000162_0001
[0330] 2-Amino-5-bromopyridine (3.0 g, 17.3 mmol) and sodium bicarbonate (2.9 g, 34.7 mmol) were stirred in dioxane (30 mL) at 100 °C and ethyl bromopyruvate (3.3 mb, 26.0 mmol) was added. The reaction was stirred for 16 h and brought to ambient temperature. It was diluted with EtOAc and washed with water, then saturated brine solution. The organic layer was separated and dried over sodium sulfate, then filtered, and concentrated in vacuo to afford the title compound (4.6 g, 99% yield). LCMS: m/z = 270.1 [M+H]+
Example 120: (6-Bromoimidazo[l,2-a]pyridin-2-yl)methanol.
Figure imgf000162_0002
[0331] Ethyl 6-bromoimidazo[l,2-a]pyridine-2-carboxylate (4.6 g, 17.1 mmol) was dissolved in THF (20 mL) under nitrogen and brought to 0 °C. DIBAL (34 mmol) was added and stirred for 1.5 h. The reaction was brought to ambient temperature and saturated Rochelle's Salt solution was added and stirred for 30 minutes. The aqueous layer was separated and extracted with EtOAc twice, and the organic layers were combined then washed with saturated brine solution. It was then dried over sodium sulfate, filtered, and dried in vacuo. The crude material was purified on silica gel in 0-10% MeOH in DCM to give the title product (1.75 g, 45% yield). LCMS: m/z = 228.0 [M+H]+. Example 121: ( 6-Bromoimidazo[ 1, 2-a]pyridin-2-yl)methoxy-tert-butyl-dimethyl-silane.
Figure imgf000163_0001
[0332] (6-Bromoimidazo[l,2-a]pyri din-2 -yl)m ethanol (1.75 g, 7.7 mmol), imidazole (682 mg, 10.0 mmol), and TBS-C1 (10.0 mmol) were stirred in DMF (20 mL) for 2 hours at room temperature. The solution was then diluted with EtOAc and washed with saturated brine solution 5 times. The organic layer was dried over sodium sulfate, filtered, and dried in vacuo. The crude material was purified on silica gel in 0-30% MTBE in hexanes to give the title product (2.65 g, 100%). LCMS: m/z = 342.9 [M+H]+.
Example 122: 2-[[Tert-butyl(dimethyl)silyl]oxymethyl]imidazo[l,2-a]pyridine-6-carbaldehyde.
Figure imgf000163_0002
[0333] (6-bromoimidazo[l,2-a]pyridin-2-yl)methoxy-tert-butyl-dimethyl-silane (195 mg, 0.57 mmol) was dissolved in dry ether (8 mL) and brought to -78 °C under nitrogen. N- butyllithium (0.57 mmol) was added and stirred for 1 h. DMF (3.5 mmol) was added and stirred for 30 min. Water was added then diluted with EtOAc. The organic layer was washed with brine solution, dried over sodium sulfate, filtered, and dried in vacuo to give the title compound (165 mg, 99%). LCMS: m/z = 291.3 [M+H]+.
Example 123: Tert-butyl-[[6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]imidazo[l,2-a]pyridin-2-yl]methoxy]-dimethyl-sdane.
Figure imgf000163_0003
[0334] (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (141 mg, 0.57 mmol), 2-[[tert-butyl(dimethyl)silyl]oxymethyl]imidazo[l,2-a]pyridine-6-carbaldehyde (165 mg, 0.57 mmol), and acetic acid (162 pL, 2.8 mmol) were dissolved in DCE (2 mL) and stirred for 2 h at 120 °C under microwave irradiation. The solution was diluted with DCM then poured into saturated sodium bicarbonate solution and the organic layer was separated. It was dried over sodium sulfate, filtered, then dried in vacuo. The crude material was purified on silica gel in 0-35% EtOAc in hexanes to give the title product (122 mg, 41% yield). LCMS: m/z = 521.6 [M+H]+.
Example 124: [6-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4, 9- tetrahydropyrido[3,4-b]indol-l-yl]inddazo[l,2-a]pyridin-2-yl]niethanoL
Figure imgf000164_0001
[0335] Tert-butyl-[[6-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]imidazo[l,2-a]pyridin-2-yl]methoxy]-dimethyl-silane (122 mg, 0.23 mmol) was dissolved in THF (3 mL) and TBAF (2.3 mmol) was added. The solution was heated to 80 °C and stirred for 2 h. The solution was cooled to room temperature then poured into a saturated solution of sodium bicarbonate. The organic layer was separated and washed with saturated brine solution 3 times. It was then dried over sodium sulfate, filtered, and dried in vacuo to yield the title product (95 mg, 100% yield). LCMS: m/z = 407.4 [M+H]+.
Example 125: 6-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4, 9- tetrahydropyrido[3,4-b]indol-l-yl]imidazo[l,2-a]pyridine-2-carbaldehyde.
Figure imgf000164_0002
[0336] [6-[(lR,3R)-2-(2-Fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]imidazo[l,2-a]pyridin-2-yl]methanol (95 mg, 0.23 mmol) and 1BX (111 mg, 1.7 mmol) were dissolved in DMSO (2 mL) and stirred for 1.5 h. The solution was diluted with EtOAc then poured into a saturated sodium bicarbonate solution and the organic layer was separated. It was washed with saturated brine solution, dried over sodium sulfate, fdtered, then dried in vacuo. The crude material was purified on silica gel in 0-30% EtOAc in hexanes to give the title product (33 mg, 35% yield). LCMS: m/z = 405.6 [M+H]+.
Example 126: (lR,3R)-l-[2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]imidazo[l,2-a]pyridin-6- yl]-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indole.
Figure imgf000165_0001
[0337] 6- [( 1 R, 3R)-2-(2-fluoro-2-methyl-propyl)-3 -methyl- 1 , 3 ,4, 9-tetrahy dropyrido[3 ,4- b]indol-l-yl]imidazo[l,2-a]pyridine-2-carbaldehyde (33 mg, 0.08 mmol) was dissolved in DMF (1 mL) and acetic acid (9.3 pL, 0.16 mmol) was added. Separately, 3 -(fluoromethyl)azeti dine hydrochloride (12.3 mg, 0.10 mmol) was dissolved in DMF (1 mL) and TEA (23 pL, 0.16 mmol) was added. These solutions were combined and stirred for 30 min. STAB (52 mg, 0.25 mmol) was added and stirred for 2 h. The reaction was quenched with MeOH (1 mL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (30.2 mg, 78% yield). LCMS: m/z = 478.6 [M+H]+. !H NMR (400 MHz, DMSO) 8 10.81 (s, 1H), 8.31 (s, 1H), 7.88 (s, 1H), 7.73 (s, 2H), 7.49 - 7.43 (m, 3H), 7.34 (d, J = 9.4 Hz, 1H), 7.29 (d, J = 8.0 Hz, 2H), 7.08 (td, J = 7.6, 1.3 Hz, 2H), 7.03 - 6.97 (m, 2H), 5.03 (s, 2H), 4.56 (d, J = 6.3 Hz, 2H), 4.44 (d, J = 6.3 Hz, 2H), 2.99 (dd, J = 7.2, 5.7 Hz, 3H), 2.86 - 2.58 (m, 5H), 1.46 (d, J = 21.8 Hz, 5H), 1.33 (s, 3H).
Example 127: 2- (hydroxyniethyl)inudazo[l, 2-a]pyridine- 7-carbonitrile
Figure imgf000165_0002
[0338] To 7-cyano-imidazo[l,2-a]pyridine-2-carboxylic acid ethyl ester (1.00 equiv, 250 mg, 1.16 mmol) in THF (10 mL) at 0 °C under nitrogen was added diisobutylaluminum hydride (2.00 equiv, 2.3 mL, 2.32 mmol) dropwise and stirred for 1 h. After slowly warming to room temp, the reaction was cooled back to 0 °C, and quenched with EtOAc, followed by saturated solution of Rochelle's salt. The mixture was stirred for 30 min. The reaction mixture was extracted with EtOAc, layers were separated, and the organic layer dried over sodium sulfate. The organic layer was fdtered, concentrated under vacuum, and purified by silica gel chromatography using 0-30% EtOAc in hexane to give title compound (70 mg, 0.404 mmol, 34.8 % yield). LCMS: m/z = 174.1 [M+H]+.
Example 128: 2-[[tert-butyl(dimethyl)silyl]oxymethyl]imidazo[l,2-a]pyridine-7-carbonitrile
Figure imgf000166_0001
[0339] To 2-(hydroxymethyl)imidazo[l,2-a]pyridine-7-carbonitrile (1.00 equiv, 80 mg, 0.462 mmol) in DMF (1 mL) at 0 °C was added imidazole (2.00 equiv, 63 mg, 0.924 mmol), followed by tert-butyldimethylchlorosilane (1.00 equiv, 70 mg, 0.462 mmol). The reaction mixture was stirred for 30 min. The reaction was quenched with water followed by brine, dried over sodium sulfate, filtered and concentrated. The crude was purified by silica gel chromatography using 0-30% EtOAc in hexane to give title compound (117 mg, 0.407 mmol, 88.1 % yield). LCMS: m/z = 288.3 [M+H]+.
Example 129: 2-(hydroxymethyl)imidazo[l,2-a]pyridine-7-carbaldehyde
Figure imgf000166_0002
[0340] To 2-[[tert-butyl(dimethyl)silyl]oxymethyl]imidazo[l,2-a]pyridine-7-carbonitrile (1.00 equiv, 117 mg, 0.407 mmol) in THF (5mL) at 0 °C under nitrogen was added diisobutylaluminum hydride (1.00 equiv, 0.41 mL, 0.407 mmol) dropwise, and the reaction mixture was stirred for 1 h. After slowly warming to room temp, the reaction was cooled back to 0 °C, and then quenched with EtOAc, followed by saturated solution of Rochelle's salt. The mixture was stirred for 30 min then extracted with EtOAc. The layers were separated, and the organic layer was dried over sodium sulfate. The organic layer was fdtered, concentrated under vacuum, and purified by silica gel chromatography using 0-30% EtOAc in DCM to give title compound (20 mg, 0.114 mmol, 27.9 % yield). LCMS: m/z = 291.3 [M+H]+.
Example 130: [7-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-
Figure imgf000167_0001
[0341] To (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (1.30 equiv, 37 mg, 0.148 mmol) in DCE (2 mL) was added 2-(hydroxymethyl)imidazo[l,2- a]pyridine-7-carbaldehyde (1.00 equiv, 20 mg, 0.114 mmol) and acetic acid (2.00 equiv, 0.013 mL, 0.227 mmol), and stirred at 110 °C in microwave for 3 h. The reaction mixture was extracted with DCM, washed with saturated sodium bicarbonate solution, dried over sodium sulfate, fdtered, and concentrated. The crude was purified by silica gel chromatography using a 24 g column elution with 0-30% EtOAc in DCM to give title compound (20 mg, 0 .0492 mmol, 43.3 % yield). LCMS: m/z = 407.2 [M+H]+.
Example 131: 7-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-
Figure imgf000167_0002
[0342] To [7-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]imidazo[l,2-a]pyridin-2-yl]methanol (1.00 equiv, 20 mg, 0.0492 mmol) in anhydrous DMSO (0.5 mL) was added 1, 1,1 -tris(acetyloxy)- 1,1 -dihydro- 1,2-benzodi oxol-3- (Ih)-one (1.20 equiv, 51 mg, 0.0590 mmol) and stirred at 25 °C for 1 h. The reaction was quenched with water and extracted with EtOAc. The EtOAc layer was washed with IN NaOH, brine, dried over sodium sulfate, filtered, and concentrated. The crude was purified by silica gel chromatography using 0-30% EtOAc in DCM to give title compound (12 mg, 0.0297 mmol, 60.3 % yield). LCMS: m/z = 405.2 [M+H]+.
Example 132: [7-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-
Figure imgf000168_0001
[0343] To 3-(fluoromethyl)azetidine hydrochloride (2.00 equiv, 7.5 mg, 0.0593 mmol) in NMP (0.5 mL) was added triethylamine (2.00 equiv, 0.0083 mL, 0.0593 mmol), stirred for 5 min, followed by addition of 7-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]imidazo[l,2-a]pyridine-2-carbaldehyde (1.00 equiv, 12 mg, 0.0297 mmol) and acetic acid (2.00 equiv, 0.0034 mL, 0.0593 mmol). The reaction mixture was stirred at room temp for 30 min, followed by addition of sodium triacetoxyborohydride (3.00 equiv, 19 mg, 0.0890 mmol) and stirred for 1 h. The reaction was quenched with methanol (0.5 mL), and purified by C-18 reverse phase chromatography using Luna column, eluting with 5- 30% ACN in water with 0.1% formic acid as modifier to give the title compound (2.0 mg, 0.00419 mmol, 14.1 % yield). LCMS: m/z = 478.4 [M+H]+.
Example 133: [2-(chloromethyl)-l,3-benzoxazol-5-yl]methanol.
Figure imgf000168_0002
[0344] Methyl 2-(chloromethyl)-l,3-benzoxazole-5- carboxylate (300 mg, 1.3 mmol) was dissolved in anhydrous THF (4 mL) under nitrogen and brought to -78 °C. DIBAL (2.0 mmol) was added and stirred for 1 h. The solution was warmed to room temperature and diluted with EtOAc. Saturated Rochelle's Salt was added and stirred for an additional hour. The layers were separated and extracted with EtOAc. The combined organic layers were washed with saturated brine solution, dried over sodium sulfate, filtered, and dried in vacuo to yield the title product (262 mg, 99% yield). LCMS: m/z = 198.5 [M+H]+. The crude was used directly in the next step.
Example 134: 2-(chloromethyl)-l,3-benzoxazole-5-carbaldehyde
Figure imgf000169_0001
[0345] [2-(Chloromethyl)-l,3-benzoxazol-5-yl]methanol (262 mg, 1.3 mmol) and IBX (742 mg, 2.6 mmol) were dissolved in DMSO (2 mL) and stirred for 2 h. The solution was diluted with EtOAc then poured into saturated sodium bicarbonate solution. The organic layer was separated and washed with saturated brine solution. It was then dried over sodium sulfate, filtered, and dried in vacuo. The crude material was purified on silica gel with 0-30% EtOAc in hexanes to give the title product (180 mg, 69% yield). LCMS: m/z = 196.6 [M+H]+.
Example 135: 2-(chloromethyl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
Figure imgf000169_0002
[0346] 2-(Chloromethyl)-l,3-benzoxazole-5-carbaldehyde (180 mg, 0.92 mmol) and (R)-N- (l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (228 mg, 0.92 mmol) were dissolved in DCE (4 mL) and acetic acid (263 pL, 4.6 mmol) was added. The solution was stirred at 120 °C under microwave irradiation for 2 h. The solution was poured into saturated sodium bicarbonate solution and the organic layer was separated. It was then washed with saturated brine solution, dried over sodium sulfate, filtered, and dried in vacuo. The crude material was purified in 0-30% EtOAc in hexanes to give the title product (140 mg, 36%). LCMS: m/z = 426.8 [M+H]+. Example 136: 2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-5-[(l R,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l, 3, 4, 9-tetrahydropyrido[3, 4-b]indol-l -yl]-l, 3-benzoxazole.
Figure imgf000170_0001
[0347] 2-(Chloromethyl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l, 3-benzoxazole (36 mg, 0.08 mmol) and 3- (fluoromethyl)azetidine hydrochloride (14 mg, 0.11 mmol) were dissolved in DMF (2 mL). TEA (47 pL, 0.34 mmol) was added and stirred for 2 h at 40 °C. The reaction was quenched with formic acid (47 pL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (14,3 mg, 35% yield), LCMS: m/z = 479.6 [M+H]+. XH NMR (400 MHz, DMSO) 6 10.82 (s, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.46 (dd, J = 4.6, 3.1 Hz, 2H), 7.36 (dd, J = 8.5, 1.7 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 7.07 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.99 (td, J = 7.4, 1.1 Hz, 1H), 5.17 (s, 1H), 4.57 (d, J = 6.2 Hz, 1H), 4.46 (d, J = 6.2 Hz, 1H), 3.83 (s, 2H), 3.43 (td, J = 7.7, 1.5 Hz, 1H), 3.12 (d, J = 6.6 Hz, 3H), 2.85 - 2.69 (m, 2H), 2.66 - 2.54 (m, 2H), 1.47 (d, J = 21.8 Hz, 3H), 1.31 (d, J = 21.5 Hz, 3H), 1.06 (d, J = 6.7 Hz, 3H).
Example 137: 2-[[3-methylazetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3- methyl-1, 3, 4, 9-tetrahydropyrido[3,4-b]indol-l-yl]-l, 3-benzoxazole.
Figure imgf000170_0002
[0348] 2-(Chloromethyl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l, 3-benzoxazole (19 mg, 0.04 mmol) and 3-methylazetidine hydrochloride (6.2 mg, 0.06 mmol) were dissolved in DMF (2 mb). TEA (25 pL, 0.18 mmol) was added and stirred for 2 h at 40 °C. The reaction was quenched with formic acid (25 pL) and directly purified by HPLC on a Kintetex 5 pm C18 100 k column (size: 100 x 30.0 mm, gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (9.8 mg, 47% yield). LCMS: m/z = 461.4 [M+H]+. XH NMR (400 MHz, DMSO) 5 10.82 (s, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.46 (d, J = 7.3 Hz, 2H), 7.36 (dd, J = 8.5, 1.7 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 7.07 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.99 (td, J = 7.5, 1.1 Hz, 1H), 5.17 (s, 1H), 3.79 (s, 2H), 3.49 - 3.44 (m, 5H), 2.87 (t, J = 6.5 Hz, 3H), 2.81 - 2.56 (m, 3H), 1.47 (d, J = 21.8 Hz, 4H), 1.31 (d, J = 21.5 Hz, 4H), 1.08 (dd, J = 19.2, 6.7 Hz, 2H).
Example 138: 2-(2-azaspiro[3.3]heptan-2-ylmethyl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)- 3-methyl-l ,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
Figure imgf000171_0002
[0349] 2-(Chloromethyl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole (30 mg, 0.07 mmol) and 2- azoniaspiro[3.3]heptane hydrochloride (612 mg, 0.09 mmol) were dissolved in DMF (2 mb). TEA (39 pL, 0.28 mmol) was added and stirred for 2 h at 40 °C. The reaction was quenched with formic acid (39 pL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (7.1 mg, 21% yield). LCMS: m/z = 487.5 [M+H]+. ’H NMR (400 MHz, DMSO) 5 10.82 (s, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.46 (d, J = 7.1 Hz, 2H), 7.35 (dd, J = 8.5, 1.7 Hz, 1H), 7 29 (d, J = 8.0 Hz, 1H), 7 07 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.99 (td, J = 7.5, 1.1 Hz, 1H), 5.17 (s, 1H), 3.76 (s, 2H), 3.25 (s, 5H), 2.85 - 2.57 (m, 3H), 2.03 (t, J = 7.6 Hz, 5H), 1.74 (pd, J = 7.3, 1.0 Hz, 2H), 1.47 (d, J = 21.8 Hz, 3H), 1.31 (d, J = 21.5 Hz, 4H), 1.06 (d, J = 6.7 Hz, 3H).
Example 139: 5-Bromo-2-(chloromethyl)-6-fluoro-l,3-benzoxazole.
Figure imgf000171_0001
[0350] 2 -Amino-4-bromo-5-fhiorophenol (1.00 equiv, 640 mg, 3.11 mmol) and ethyl 2- chloroethanimidoate hydrochloride (1.30 equiv, 638 mg, 4.04 mmol) were heated at 90 °C in ethanol (10 mL) overnight. The volatiles were removed under reduced pressure, and the crude was dissolved in DCM. The insoluble material was filtered, and the solution was concentrated under reduced pressure to give the title product (783 mg, 95% yield). LCMS: m/z = 265.5 [M+H]+.
Example 140: 5-Bromo-6-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole.
Figure imgf000172_0001
[0351] 3-(Fluoromethyl)azetidine hydrochloride (1.20 equiv, 446 mg, 3.55 mmol) and 5- bromo-2-(chloromethyl)-6-fluoro-l,3-benzoxazole (1.00 equiv, 783 mg, 2.96 mmol) were dissolved in DCM (15 mL), warmed to 50 °C, and stirred for 1.5 h. The solution was then concentrated under reduced pressure. The crude was purified on silica gel in 0-10% MeOH (2% TEA) in DCM to give the title product (725 mg, 77% yield). LCMS: m/z = 318.1 [M+H]+.
Example 141: 6-Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5- carbonitrile.
Figure imgf000172_0002
[0352] 5 -Bromo-6-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole (1.00 equiv, 460 mg, 1.45 mmol), Xantphos Pd G4 (0.0800 equiv, 112 mg, 0.116 mmol), zinc (0.200 equiv, 19 mg, 0.290 mmol) and zinc cyanide (1.00 equiv, 170 mg, 1.45 mmol) were dissolved in dry NMP (4 mL) and stirred under microwave irradiation for 10 min at 100 °C. The mixture was diluted with EtOAc and washed with saturated brine solution 3 times. The organic layers were dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0- 10% MeOH (2% TEA) in DCM to give the title product (346 mg, 91% yield). LCMS: m/z = 264.3 [M+H]+. Example 142: 6-Eluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5- carbaldehyde.
Figure imgf000173_0001
[0353] 6 -Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5-carbonitrile
(1.00 equiv, 346 mg, 1.31 mmol) was dissolved in dry THF (10 mL) under nitrogen and cooled to 0 °C. DIBAL (4 mL, 1 M in toluene) was added dropwise and stirred for 1 h. The reaction mixture was then warmed to room temperature and quenched with EtOAc. Rochelle's Salt solution was then added and stirred for 30 min. The organic layers were then separated, dried over sodium sulfate, fdtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH (2% TEA) in DCM to give the title product (55 mg, 16% yield). LCMS: m/z = 267.3 [M+H]+.
Example 143: 6-Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2- methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
Figure imgf000173_0002
[0354] (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (1.07 equiv, 55 mg, 0.221 mmol) and 6-fhjoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5- carbaldehyde (1.00 equiv, 55 mg, 0.207 mmol) were dissolved in DCE (3 mL), and acetic acid (5.00 equiv, 0.059 mL, 1.03 mmol) was added. The solution was stirred under microwave irradiation for 2 h at 120 °C. The solution was diluted with DCM, poured into saturated sodium bicarbonate solution, and the organic layers were separated. The organic layers were dried over sodium sulfate, filtered, and dried in vacuo. The crude was dissolved in DMF (3 mL) and was purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (7.7 mg, 7% yield). LCMS: m/z = 497.6 [M+H]+; XH NMR (400 MHz, DMSO) 8 10.66 (s, 1H), 7.46 (d, J = 7.7 Hz, 1H), 7.29 - 7.18 (m, 1H), 7.02 (dtd, J = 23.5, 7.1, 1.2 Hz, 2H), 6.72 (d, J = 10.5 Hz, 1H), 6.10 (d, J = 7.9 Hz, 1H), 5.28 (s, 1H), 4.59 (d, J = 5.7 Hz, 1H), 4.48 (d, J = 7.8 Hz, 2H), 4.04 (s, 2H), 3.76 (s, 4H), 3.08 - 2.61 (m, 4H), 1.40 (d, J = 21.6 Hz, 3H), 1.26 (d, J = 21.2 Hz, 3H), 1.05 (d, J = 6.6 Hz, 3H).
Example 144: 5-Bromo-2-(chloromethyl)-7-fluoro-l,3-benzoxazole.
Figure imgf000174_0001
[0355] 2 -Amino-4-bromo-6-fluoro-phenol (1.00 equiv, 640 mg, 3.11 mmol) and ethyl 2- chloroethanimidoate hydrochloride (1.30 equiv, 638 mg, 4.04 mmol) were heated at 90 °C in ethanol (10 mL) overnight. The volatiles were removed under reduced pressure, and the crude was dissolved in DCM. The insoluble material was filtered, and the solution was concentrated under reduced pressure to give the title product (821 mg, 100% yield). LCMS: m/z = 265.5 [M+H]+.
Example 145: 5-Bromo-7-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole.
Figure imgf000174_0002
[0356] 3-(Fluoromethyl)azetidine hydrochloride (1.20 equiv, 468 mg, 3.73 mmol) and 5- bromo-2-(chloromethyl)-7-fluoro-l,3-benzoxazole (1.00 equiv, 821 mg, 3.10 mmol) were dissolved in DCM (15 mL) and warmed to 50 °C and stirred for 1.5 h. The solution was then concentrated under reduced pressure. The crude material was purified on silica gel in 0-10% MeOH (2% TEA) in DCM to give the title product (845 mg, 86% yield). LCMS: m/z = 318.1 [M+H]+. Example 146: 7-Eluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]- 1 ,3-benzoxazole-5- carbonitrile.
Figure imgf000175_0001
[0357] l,l'-Bis(diphenylphosphino)ferrocene-palladium(II) di chloride di chloromethane complex (0.100 equiv, 188 mg, 0.257 mmol), 5-bromo-7-fluoro-2-[[3-(fluoromethyl)azetidin-l- yl]methyl]-l,3-benzoxazole (1.00 equiv, 815 mg, 2.57 mmol), zinc (0.250 equiv, 42 mg, 0.642 mmol), zinc cyanide (2.00 equiv, 603 mg, 5.14 mmol), and tris(dibenzylideneacetone)dipalladium(0) (0.0500 equiv, 118 mg, 0.128 mmol) were dissolved in dry DMF (10 mL) and stirred at 120 °C for 3 h. The mixture was then cooled to room temperature and diluted with EtOAc. It was then filtered and poured into saturated brine solution. The organic layers were then separated and washed with saturated brine solution 5 times. The organic layers were dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH (2% TEA) in DCM to give the title product (508 mg, 75% yield). LCMS: m/z = 264.2 [M+H]+.
Example 147: 7-Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5- carbaldehyde.
Figure imgf000175_0002
[0358] 7 -Fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5-carbonitrile
(1.00 equiv, 508 mg, 1.93 mmol) was dissolved in dry THF (8 mL) under nitrogen and cooled to 0 °C . DIBAL (5.7 mL, 1 M in toluene) was added dropwise and stirred for 1 h. The reaction was then warmed to room temperature and quenched with EtOAc. Rochelle's Salt solution was then added and stirred for 30 min. The organic layers were then separated, dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH (2% TEA) in DCM to give the title product. LCMS: m/z = 267.3 [M+H]+. Example 148: 7-Eluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2- methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
Figure imgf000176_0001
[0359] (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (1.20 equiv, 62 mg, 0.248 mmol) and 7-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5- carbaldehyde (1.00 equiv, 55 mg, 0207 mmol) were dissolved in DCE (3 mL), and acetic acid (5.00 equiv, 62.0 mg, 1.03 mmol) was added. The solution was stirred under microwave irradiation for 2 h at 120 °C. The solution was diluted with DCM, poured into saturated sodium bicarbonate solution, and the organic layers were separated. The organic layers were dried over sodium sulfate, filtered, and dried in vacuo. The crude was dissolved in DMF (2 mL) and was directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (115.9 mg, 55% yield). LCMS: m/z = 497.6 [M+H]+; rH NMR (400 MHz, DMSO) 8 10.82 (s, 1H), 7.44 (dd, J = 7.7, 3.6 Hz, 1H), 7.36 - 7.25 (m, 1H), 7.12 - 7.01 (m, 1H), 6.98 (ddd, J = 8.0, 7.0, 1.2 Hz, 1H), 6.80 - 6.66 (m, 1H), 6.51 (d, J = 1.9 Hz, 1H), 4.94 (s, 1H), 4.69 - 4.60 (m, 1H), 4.53 (d, J = 10.5 Hz, 3H), 3.99 - 3.64 (m, 1H), 3.20 - 2.89 (m, 1H), 2.82 - 2.54 (m, 2H), 2.49 - 2.38 (m, 1H), 1.47 (dd, J = 21.8, 4.9 Hz, 3H), 1.30 (d, J = 21.5 Hz, 3H), 1.08 (dd, J = 6.7, 4.5 Hz, 3H).
Example 149: Tert-butyl-[3-[(lR,3R)-l-[2-(chloromethyl)-l,3-benzoxazol-5-yl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]-2,2-difluoro-propoxy]-diphenyl-silane.
Figure imgf000176_0002
[0360] 2-(Chlorom ethyl)- 1,3 -benzoxazole-5-carbaldehyde (85 mg, 0.43 mmol) and (R)-N- ( 1 -( 1 H-indol-3 -yl)propan-2-yl)-3 -((tert-butyl diphenyl silyl)oxy)-2,2-difluoropropan- 1 -amine (264 mg, 0.52 mmol) were dissolved in DCE (4 mL), and acetic acid (124 pL, 2.2 mmol) was added. The solution was stirred at 120 °C under microwave irradiation for 2 h. The solution was poured into saturated sodium bicarbonate solution and the organic layer was separated. It was then washed with saturated brine solution, dried over sodium sulfate, fdtered, and dried in vacuo. The crude material was purified via flash chromatography on silica gel in 0-30% EtOAc in hexanes to give the title product (218 mg, 73%). LCMS: m/z = 685.2 [M+H]+.
Example 150: 3-[(lR,3R)-l-[2-(Chloromethyl)-l,3-benzoxazol-5-yl]-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-2-yl]-2,2-difluoro-propan-l-ol.
Figure imgf000177_0001
[0361] Tert-butyl-[3-[(lR, 3R)-l-[2-(chl oromethyl)-!, 3-benzoxazol-5-yl]-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-2-yl]-2,2-difluoro-propoxy]-diphenyl-silane (218 mg, 0.32 mmol) was dissolved in THF (3 mL), and TBAF (1.0 M in THF, 1 mL) was added and stirred for 30 min. The solution was diluted with EtOAc and poured into saturated sodium bicarbonate solution. The organic layers were separated and washed with saturated brine solution. The organic layers were then dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified via flash chromatography on silica gel in 0-30% EtOAc in hexanes to give the title product (86 mg, 61% yield). LCMS: m/z = 447.0 [M+H]+.
Example 151: 2,2-Difluoro-3-[(lR,3R)-l-[2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3- benzoxazol-5-yl]-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-l-ol.
Figure imgf000178_0001
[0362] 3-[(lR,3R)-l-[2-(Chloromethyl)-l,3-benzoxazol-5-yl]-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-2-yl]-2,2-difluoro-propan-l-ol (86 mg, 0.19 mmol) and 3- (fluoromethyl)azetidine hydrochloride (36 mg, 0.29 mmol) were dissolved in DMF (2 mL). TEA (108 pL, 0.77 mmol) was added and stirred for 2 h at 40 °C. The reaction was quenched with formic acid (80 pL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water), then lyophilized to give the title compound as a white solid (28.7 mg, 29% yield). LCMS: m/z = 499.6 [M+H]+; ’H NMR (400 MHz, DMSO) 5 10.87 (s, 1H), 7.65 (d, J = 8.5 Hz, 1H), 7.52 - 7.41 (m, 2H), 7.36 - 7.26 (m, 2H), 7.13 - 7.07 (m, 1H), 7.07 - 6.97 (m, 1H), 5.16 (s, 1H), 4.58 (d, J = 6.2 Hz, 1H), 4.46 (d, J = 6.2 Hz, 1H), 3.83 (s, 2H), 3.76 (td, J = 14.1, 4.0 Hz, 2H), 3.43 (td, J = 7.7, 1.5 Hz, 3H), 3.27 - 3.05 (m, 5H), 2.81 - 2.61 (m, 3H), 2.57 (d, J = 10.0 Hz, 1H), 1.08 (d, J = 6.7 Hz, 3H).
Example 152: Tert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[7-fluoro-2-[[3-(fluoromethyl)azetidin-l- yl]methyl]-l ,3-benzoxazol-5-yl]-3-methyl-l ,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propoxy]- diphenyl-silane.
Figure imgf000178_0002
[0363] (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2- difluoropropan- 1 -amine (1.20 equiv, 126 mg, 0.248 mmol) and 7-fluoro-2-[[3-
(fhioromethyl)azetidin-l-yl]methyl]-l,3-benzoxazole-5-carbaldehyde (1.00 equiv, 55 mg, 0.207 mmol) were dissolved in DCE (3 mL), and acetic acid (5.00 equiv, 0.059 mL, 1.03 mmol) was added. The solution was stirred under microwave irradiation for 2 h at 120 °C. The solution was diluted with DCM, poured into saturated sodium bicarbonate solution, and the organic layers were separated. The organic layers were dried over sodium sulfate, fdtered, and dried in vacuo. The crude (210 mg, 135% yield) was used directly in the next step.
Example 153: 2,2-Difluoro-3-[(lR,3R)-l-[7-fluoro-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]~ l,3-benzoxazol-5-yl]-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-L-ol.
Figure imgf000179_0001
[0364] Tert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[7-fluoro-2-[[3-(fluoromethyl)azetidin-l- yl]methyl]-l,3-benzoxazol-5-yl]-3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propoxy]- diphenyl-silane (210 mg, 0.28 mmol, 45% pure) was dissolved in THF (5 mL) and TBAF (2.00 equiv, 0.556 mmol) in THF was added and stirred for 30 min. The solution was diluted in EtOAc and poured into saturated sodium bicarbonate solution. The organic layers were washed with brine solution, dried over sodium sulfate, fdtered, and dried in vacuo. The crude was dissolved in DMF (2 mL) and was purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (7.2 mg, 11% yield). LCMS: m/z = 517.4 [M+H]+. 'l l NMR (400 MHz, DMSO) 8 10.87 (s, 1H), 7.45 (d, J = 7.7 Hz, 1H), 7.30 (d, J = 7.9 Hz, 1H), 7.08 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 6.99 (td, J = 7.5, 1.1 Hz, 1H), 6.76 - 6.58 (m, 1H), 6.54 - 6.43 (m, 1H), 4.93 (s, 1H), 4.69 - 4.58 (m, 1H), 4.53 (d, J = 12.4 Hz, 3H), 3.74 (t, J = 13.7 Hz, 2H), 3.24 - 2.93 (m, 4H), 2.73 - 2.54 (m, 2H), 1.09 (d, J = 6.7 Hz, 3H). Example 154: Tert-butyl 3-[(5-bromo-2-hydroxy-phenyl)carbamoyl]azetidine-l-carboxylate. Boc
Figure imgf000180_0001
[0365] 2-Amino-4-bromophenol (1.00 equiv, 2.00 g, 10.6 mmol), l-Boc-azetidine-3- carboxylic acid (1.00 equiv, 2.14 g, 10.6 mmol), and HATU (1.00 equiv, 4.04 g, 10.6 mmol) were dissolved in DMF (16 mb). N,N-Diisopropylethylamine (3.00 equiv, 5.6 mb, 31.9 mmol) was added, and the reaction mixture was stirred for 1.5 h then diluted with EtOAc. The solution was then poured into saturated brine solution, and the organic layers were separated and washed with brine 5 times. The organic layers were then dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-30% EtOAc in DCM to give the desired product (2.96 g, 75% yield). LCMS: m/z = 372.2 [M+H]+.
Example 155: Tert-butyl 3-(5-bromo-l,3-benzoxazol-2-yl)azetidine-l-carboxylate.
Figure imgf000180_0002
[0366] Tert-butyl 3-[(5-bromo-2-hydroxy-phenyl)carbamoyl]azetidine-l-carboxylate (1.00 equiv, 2.96 g, 7.97 mmol) and triphenylphosphine (1.10 equiv, 2301 mg, 8.77 mmol) were dissolved in dry THF (15 mL) under nitrogen and stirred at room temperature. DIAD (1.10 equiv, 1.7 mL, 8.77 mmol) was added dropwise and stirred for 1.5 h at room temperature. The solution was then dried in vacuo and purified on silica gel in 0-30% EtOAc in hexanes to give the desired product (2.86 g, 102% yield). LCMS: m/z = 354.2 [M+H]+.
Example 156: Tert-butyl 3-(5-cyano-l,3-benzoxazol-2-yl)azetidine-l-carboxylate.
Figure imgf000180_0003
[0367] Tert-butyl 3-(5-bromo-l,3-benzoxazol-2-yl)azetidine-l-carboxylate (1.00 equiv, 536 mg, 1.52 mmol), tetrakis(triphenylphosphine)palladium(0) (0.0500 equiv, 88 mg, 0.0759 mmol), and zinc cyanide (1.00 equiv, 178 mg, 1.52 mmol) were dissolved in dry DMF and under an atmosphere of nitrogen and stirred at 80 °C overnight. The solution was cooled, diluted with EtOAc, and poured into water. The organic layer was washed with brine 5 times, dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on a 24 g silica column in 0- 40% EtOAc in hexanes to give the desired product (238 mg, 52% yield). LCMS: m/z = 300.3 [M+H]+.
Example 157: Tert-butyl 3-(5-formyl-l,3-benzoxazol-2-yl)azetidine-l-carboxylate.
Figure imgf000181_0001
[0368] Tert-butyl 3-(5-cyano-l,3-benzoxazol-2-yl)azetidine-l-carboxylate (1.00 equiv, 238 mg, 0.795 mmol) was dissolved in dry THF (8 mL) under nitrogen and cooled to 0 °C . DIBAL (1 M in toluene, 1 mL) was added dropwise and stirred for 1 h. The reaction was then warmed to room temperature and quenched with EtOAc. Rochelle's Salt solution was then added and stirred for 30 min. The organic layers were then separated, dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-40% EtOAc in hexanes to give the title product (38 mg, 16%). LCMS: m/z = 303.4 [M+H]+.
Example 158: Tert-butyl 3-[5-[(lR,3R)-2-(2-fluoro-2-metliyl-propyl)-3-metliyl-l,3,4,9- tetrahydropyrido[3, 4-b]indol-l-yl]-l, 3-b enzoxazol-2-yl] azetidine- 1-carboxy late.
Figure imgf000181_0002
[0369] (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (1.00 equiv,
31 mg, 0.126 mmol) and tert-butyl 3-(5-formyl-l ,3-benzoxazol-2-yl)azetidine-l -carboxylate (1.00 equiv, 38 mg, 0.126 mmol) were dissolved in DCE (1.5 mL), and acetic acid (5.00 equiv, 0.036 mL, 0.628 mmol) was added. The solution was stirred under microwave irradiation for 2 h at 120 °C. The solution was diluted with DCM, poured into saturated sodium bicarbonate solution, and the organic layers were separated. The organic layers were dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-30% EtOAc in hexanes to give the title product (39 mg, 58% yield). LCMS: m/z = 533.6 [M+H]+.
Example 159: 2-(Azetidin-3-yl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole.
Figure imgf000182_0001
[0370] Tert-butyl 3-[5-[(lR, 3R)-2-(2-fluoro-2-methyl-propyl)-3 -methyl- 1, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazol-2-yl]azetidine-l-carboxylate (1.00 equiv, 39 mg, 0.0732 mmol) was dissolved in DCM and cooled to 0 °C . TFA (40.0 equiv, 0.23 mb, 2.93 mmol) was added dropwise and stirred for 2 h. The volatiles were removed under reduced pressure then azeotroped with toluene. The crude (29 mg, 92% yield) was used directly in the next step. LCMS: m/z = 433.6 [M+H]+.
Example 160: 5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4-
Figure imgf000182_0002
[0371] 2-(Azetidin-3-yl)-5-[(lR,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzoxazole (1.00 equiv, 16 mg, 0.0370 mmol) was dissolved in DMF. l-Bromo-3 -fluoropropane (1.00 equiv, 5.2 mg, 0.0370 mmol) and triethylamine (3.00 equiv, 0.015 mL, 0.111 mmol) were added. The solution was stirred for 1.5 h. The reaction was directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water), then lyophilized to give the title compound as a white solid (6.1 mg, 28% yield). LCMS: m/z = 493.6 [M+H]+; XH NMR (400 MHz, CDCh) 5 7.63 (s, 2H), 7.59 - 7.50 (m, 1H), 7.46 - 7.35 (m, 2H), 7.31 (s, 1H), 7.17 (dtd, J = 17.5, 7.1, 1.3 Hz, 2H), 5.21 (s, 1H), 4.58 (t, J = 5.8 Hz, 1H), 4.46 (t, J = 5.8 Hz, 1H), 4.14 - 3.95 (m, 3H), 3.64 (t, J = 7.4 Hz, 2H), 2.85 - 2.55 (m, 5H), 1.86 (dq, J = 26.3, 6.5 Hz, 2H), 1.45 (d, J = 4.6 Hz, 3H), 1.36 (s, 2H), 1.29 (d, J = 8.6 Hz, 3H), 1.10 (d, J = 6.7 Hz, 3H).
Example 161: 2-[l-[[5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]azetidin-3-yl]propan-2-ol.
Figure imgf000183_0001
[0372] 5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 2-(azeti din-3 -yl)propan-2-ol (6 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm, gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (4.2 mg, 17% yield). LCMS: m/z = 503.7 [M+H]“. Example 162: 6-(Difluoromethyl)-2-[[5-[(lR)-2-(2-fluoro-2-metliyl-propyl)-3,3-dimetliyl-4,9- dihydro-lH-pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]-2-azaspiro[3.3]heptan-6-ol.
Figure imgf000184_0001
[0373] 5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 6-(difluoromethyl)-2- azaspiro[3.3]heptan-6-ol hydrochloride (10 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size. 100 x 30.0 mm; gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (12.6 mg, 47% yield). LCMS: m/z = 551.3 [M+H]+.
Example 163: l-[[5-[(lR)-2-(2-Fluoro-2-tnethyl-propyl)-3,3-dimethyl-4,9-diliydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]azetidine-3-carbonitrile.
Figure imgf000184_0002
[0374] 5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4- b]indol-l -yl]-lH-indole-2-carbaldehyde (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 3 -cyanoazetidine hydrochloride (6 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (4.9 mg, 21% yield). LCMS: m/z = 470.2 [M+H]“.
Example 164: (lR)-2-(2-Fluoro-2-methyl-propyl)-l-[2-[[3-(methoxymethyl)azetidin-l- yl]methyl]-lH-indol-5-yl]-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4-b]indole.
Figure imgf000185_0001
[0375] 5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 3 -(methoxymethyl)-azetidine hydrochloride (7 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm, gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (4.8 mg, 20% yield). LCMS: m/z = 489.7 [M+H]+.
Example 165: (lR)-l-[2-[(6-Fluoro-2-azaspiro[3.3]heptan-2-yl)metliyl]-lH-indol-5-yl]-2-(2- fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4-b]indole.
Figure imgf000185_0002
[0376] 5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 6-fluoro-2- azoniaspiro[3.3]heptane;2,2,2-trifluoroacetate (11 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm Cl 8 100 A column (size: 100 x 30.0 mm; gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (7.1 mg, 29% yield). LCMS: m/z = 503.6 [M+H]+.
Example 166: 2-[[5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]-2-azaspiro[3.3]heptan-6-oL
Figure imgf000186_0001
[0377] 5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 2-azaspiro[3.3]heptan-6-ol hydrochloride (7 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (11.6 mg, 47% yield). LCMS: m/z = 501.5 [M+H]+.
Example 167: 6-[[5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH- pyrido[3,4-b]indol-l-yl]-lH-indol-2-yl]methyl]-2-oxa-6-azaspiro[3.3]heptane.
Figure imgf000186_0002
[0378] 5-[(lR)-2-(2-Fluoro-2-methyl-propyl)-3,3-dimethyl-4,9-dihydro-lH-pyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (20 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and acetic acid (6 pL, 0.10 mmol) was added. Separately, 2-oxa-6-azaspiro[3.3]heptane; oxalic acid (9 mg, 0.05 mmol) was dissolved in DMF (0.5 mL) and TEA (14 pL, 0.10 mmol) was added. These solutions were combined and stirred for 30 minutes. STAB (32 mg, 0.15 mmol) was added and stirred for 2 hours. The reaction was quenched with 1 mL MeOH and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-40% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as an off-white solid (9.9 mg, 41 % yield). LCMS: m/z = 487.3 [M+H]+.
Example 168: (lR,3R)-2-(2-Fluoro-2-methylpropyl)-l-(2-((3-fluoroazetidin-l-yl)methyl)-lH-
Figure imgf000187_0001
[0379] A solution of 5-((17?,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- l/7-pyrido[3,4-Z>]indol-l-yl)-lJH-indole-2-carbaldehyde (60 mg, 0.15 mmol, 1 equiv) in di chloromethane (1.9 mL) was added to a mixture of 3 -fluoroazetidine hydrochloride salt (33 mg, 0.30 mmol, 2 equiv) and triethylamine (0.04 mL, 0.3 mmol, 2 equiv) in di chloromethane (1.9 mL). Acetic acid (0.02 mL, 0.3 mmol, 2 equiv) was added to the resulting reaction mixture which was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (0.095 g, 0.45 mmol, 3 equiv) was added and reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with di chloromethane (15 mL) and washed with saturated sodium carbonate (15 mL). The aqueous layer was extracted with di chloromethane (2 x 15 mL). The combined organic layers were dried over sodium sulfate, fdtered and concentrated under reduced pressure onto Celite® (1 g). The residue was purified on a Biotage automated chromatography system (5.5 g, Redigold C18 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound (46 mg, 65% yield) as an off-white solid. LCMS: m/z = 463.3 [M+H]+; XH NMR (400 MHz, CDCk) 8 = 8.33 (br s, 1H), 7.67 - 7.43 (m, 2H), 7.31 (s, 1H), 7.29 - 7.26 (m, 1H), 7.25 - 7.19 (m, 2H), 7.19 - 7.07 (m, 2H), 6.25 (s, 1H), 5.20 (quin, J = 5.2 Hz, 1H), 5.13 - 5.02 (m, 1H), 3.78 (s, 2H), 3.70 - 3.57 (m, 2H), 3.53 (br s, 1H), 3.34 - 3.17 (m, 2H), 2.86 (br d, J= 13.0 Hz, 1H), 2.80 - 2.54 (m, 3H), 1.43 (br s, 1H), 1.40 - 1.31 (m, 3H), 1.31 - 1.17 (m, 2H), 1.08 (d, J= 6.7 Hz, 3H).
Example 169: (lR,3R)-2-(2-Fluoro-2-methylpropyl)-l-(2-((3-methoxyazetidin-l-yl)methyl)~
Figure imgf000188_0001
[0380] A solution of 5-((17?,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- 12/-pyrido[3,4-Z>]indol-l-yl)-l/7-indole-2-carbaldehyde (60 mg, 0.15 mmol, 1 equiv) in di chloromethane (1.9 mL) was added to a mixture of 3-methoxyazetidine hydrochloride salt (37 mg, 0.30 mmol, 2 equiv) and triethylamine (0.04 mL, 0.3 mmol, 2 equiv) in di chloromethane (1.9 mL). Acetic acid (0.02 mL, 0.3 mmol, 2 equiv) was added to the resulting reaction mixture which was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (0.095 g, 0.45 mmol, 3 equiv) was added and reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with di chloromethane (15 mL) and washed with saturated sodium carbonate (15 mL). The aqueous layer was extracted with dichloromethane (2 x 15 mL). The combined organic layers were dried over sodium sulfate, fdtered and concentrated under reduced pressure onto Celite® (1 g). The residue was purified on a Biotage automated chromatography system (5.5 g, Redigold C18 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound (29 mg, 41% yield) as an off-white solid. LCMS: m/z = 475.3 [M+H]+; 'H NMR (400 MHz, CDCh) 8 = 8.44 (br s, 1H), 7.63 - 7.52 (m, 2H), 7.32 (d, J = 4.5 Hz, 1H), 7.25 - 7.21 (m, 3H), 7.17 - 7.10 (m, 2H), 6.24 (s, 1H), 5.08 (br s, 1H), 4.05 (quin, J =
5.6 Hz, 1H), 3.76 (s, 2H), 3.62 - 3.52 (m, 3H), 3.26 (s, 3H), 3.04 (dd, J = 5.4, 7.3 Hz, 2H), 2.87 (br d, J= 14.5 Hz, 1H), 2.77 - 2.56 (m, 3H), 1.46 - 1.35 (m, 3H), 1.34 - 1.28 (m, 3H), 1.09 (d, J =
6.7 Hz, 3H). Example 170: (lR,3R)-l-(2-((3-Ethylazetidin-l-yl)methyl)-lH-indol-5-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole
Figure imgf000189_0001
[0381] A solution of 5-((17?,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- l/f-pyrido[3,4-/>]indol-l-yl)-177-indole-2-carbaldehyde (58 mg, 0.14 mmol, 1 equiv) in N,N- dimethylformamide (1.3 mL) was added to a mixture of 3 -ethylazetidine trifluoroacetic acid salt (57 mg, 0.29 mmol, 2 equiv) and triethylamine (0.04 mL, 0.3 mmol, 2 equiv) in N,N- dimethylformamide (1.3 mL). Acetic acid (0.02 mL, 0.3 mmol, 2 equiv) was added to the resulting reaction mixture which was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (0.092 g, 0.43 mmol, 3 equiv) was added and reaction was stirred at room temperature for 3 days. The reaction mixture was diluted with ethyl acetate (15 mL) and washed with saturated sodium bicarbonate (15 mL). The organic layer was sequentially washed with water (2 x 15 mL) and saturated brine (15 mL). The organic layer was dried over sodium sulfate, fdtered and concentrated under reduced pressure onto Celite® (0.8 g). The residue was purified on a Biotage automated chromatography system (11 g, Biotage 50 pm KP-amino-d column), eluting with a gradient of 0 to 100% ethyl acetate in hexanes to give the title compound (50 mg, -85% purity). The resulting residue was loaded onto Celite® (1 g) purified on a Biotage automated chromatography system (5.5 g Redigold C18 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound (21 mg, 31% yield) a pale-yellow solid. LCMS: m/z = 473.3 [M+H]+; 'H NMR (400 MHz, CDCk) 8 = 8.53 (br s, 1H), 7.64 - 7.44 (m, 2H), 7.32 - 7.27 (m, 1H), 7.24 - 7.20 (m, 3H), 7.17 - 7.07 (m, 2H), 6.21 (s, 1H), 5.05 (br s, 1H), 3.69 (s, 2H), 3.55 (br s, 1H), 3.43 (t, J = 7.5 Hz, 2H), 2.93 - 2.74 (m, 3H), 2.74 - 2.62 (m, 1H), 2.62 - 2.54 (m, 2H), 2.38 (spt, J= 7.2 Hz, 1H), 1.53 (quin, J= 7.4 Hz, 2H), 1.40 (br s, 1H), 1.38 - 1.30 (m, 3H), 1.27 (s, 2H), 1.07 (d, J = 6.6 Hz, 3H), 0.81 (t, J= 7.4 Hz, 3H). Example 171 : 2-(l-((5-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indol-2-yl)methyl)azetidin-3-yl)acetonitrile
Figure imgf000190_0001
[0382] A solution of 5-((17?,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- l/7-pyrido[3,4-/>]indol-l-yl)-l/7-indole-2-carbaldehyde (60 mg, 0.15 mmol, 1 equiv) in di chloromethane (1.9 mL) was added to a mixture of 2-(azeti din-3 -yl)acetonitrile hydrochloride salt (40. mg, 0.30 mmol, 2 equiv) and triethylamine (0.04 mL, 0.3 mmol, 2 equiv) in di chloromethane (1.9 mL). Acetic acid (0.02 mL, 0.3 mmol, 2 equiv) was added to the resulting reaction mixture which was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (0.095 g, 0.45 mmol, 3 equiv) was added and reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with dichloromethane (15 mL) and washed with saturated sodium carbonate (15 mL). The aqueous layer was extracted with di chloromethane (2 x 15 mL). The combined organic layers were dried over sodium sulfate, fdtered and concentrated under reduced pressure onto Celite® (1 g). The residue was purified on a Biotage automated chromatography system (5.5 g, Redigold Cl 8 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound (42 mg, 58% yield) as an off-white solid. LCMS: m/z = 484.3 [M+H]+; ’H NMR (400 MHz, CDCk) 8 = 8.30 (br s, 1H), 7.65 - 7.45 (m, 2H), 7.31 (s, 1H), 7.25 - 7.20 (m, 3H), 7.17 - 7.09 (m, 2H), 6.23 (s, 1H), 5.07 (br s, 1H), 3.72 (s, 2H), 3.53 (br s, 1H), 3.44 (t, J= 7.6 Hz, 2H), 3.08 - 3.00 (m, 2H), 2.89 - 2.69 (m, 3H), 2.67 - 2.55 (m, 4H), 1.42 (br s, 1H), 1.40 - 1.31 (m, 3H), 1.30 - 1.17 (m, 2H), 1.08 (d, J = 6.6 Hz, 3H). Example 172: (lR,3R)-2-(2-Fluoro-2-methylpropyl)-l-(2-((3-isopropylazetidin-l-yl)methyl)-
Figure imgf000191_0001
[0383] A solution of 5-((17?,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- l/f-pyrido[3,4-/>]indol-l-yl)-177-indole-2-carbaldehyde (60 mg, 0.15 mmol, 1 equiv) in di chloromethane (1.9 mL) was added to a mixture of 3-isopropylazetidine hydrochloride salt (40. mg, 0.30 mmol, 2 equiv) and triethylamine (0.04 mL, 0.3 mmol, 2 equiv) in di chloromethane (1.9 mL). Acetic acid (0.02 mL, 0.3 mmol, 2 equiv) was added to the resulting reaction mixture which was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (0.095 g, 0.45 mmol, 3 equiv) was added and reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with di chloromethane (15 mL) and washed with saturated sodium carbonate (15 mL). The aqueous layer was extracted with di chloromethane (2 x 15 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure onto Celite® (1 g). The residue was purified on a Biotage automated chromatography system (5.5 g, Redigold C18 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound (35 mg, 48% yield) as an off-white solid. LCMS: m/z = 487.3 [M+H]+; LH NMR (400 MHz, CDCh) S = 8.61 (br s, 1H), 7.63 - 7.46 (m, 2H), 7.33 - 7.27 (m, 1H), 7.23 - 7.08 (m, 5H), 6.21 (s, 1H), 5.05 (br s, 1H), 3.67 (s, 2H), 3.54 (br s, 1H), 3.47 - 3.40 (m, 2H), 2.89 - 2.75 (m, 3H), 2.72 - 2.54 (m, 3H), 2.18 - 2.08 (m, 1H), 1.72 - 1.59 (m, 1H), 1.44 - 1.37 (m, 1H), 1.37 - 1.30 (m, 3H), 1.29 - 1.13 (m, 2H), 1.07 (d, J= 6.1 Hz, 3H), 0.79 (d, J= 6.6 Hz, 6H). Example 173 : (lR,3R)-l-(2-((3-(Difluoromethyl)azetidin-l-yl)methyl)-lH-indol-5-yl)-2-(2- fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole
Figure imgf000192_0001
[0384] A solution of 5-((17?,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- l/7-pyrido[3,4-/>]indol-l-yl)-l/7-indole-2-carbaldehyde (60 mg, 0.15 mmol, 1 equiv) in di chloromethane (1.9 mL) was added to a mixture of 3-(difluoromethyl)azetidine hydrochloride salt (43 mg, 0.30 mmol, 2 equiv) and triethylamine (0.04 mL, 0.3 mmol, 2 equiv) in di chloromethane (1.9 mL). Acetic acid (0.02 mL, 0.3 mmol, 2 equiv) was added to the resulting reaction mixture which was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (0.095 g, 0.45 mmol, 3 equiv) was added and reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with dichloromethane (15 mL) and washed with saturated sodium carbonate (15 mL). The aqueous layer was extracted with di chloromethane (2 x 15 mL). The combined organic layers were dried over sodium sulfate, fdtered and concentrated under reduced pressure onto Celite® (1 g). The residue was purified on a Biotage automated chromatography system (5.5 g, Redigold Cl 8 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound (45 mg, 61% yield) as an off-white solid. LCMS: m/z = 495.3 [M+H]+; ’H NMR (400 MHz, CDCk) 8 = 8.31 (br s, 1H), 7.61 - 7.48 (m, 2H), 7.31 (s, 1H), 7.26 - 7.20 (m, 3H), 7.17 - 7.09 (m, 2H), 6.24 (s, 1H), 6.14 - 5.81 (m, 1H), 5.08 (br s, 1H), 3.71 (s, 2H), 3.54 (br s, 1H), 3.41 - 3.32 (m, 2H), 3.26 - 3.19 (m, 2H), 2.92 - 2.76 (m, 2H), 2.73 - 2.55 (m, 3H), 1.43 (br s, 1H), 1.40 - 1.32 (m, 3H), 1.31 - 1.24 (m, 2H), 1.08 (d, J= 6.6 Hz, 3H). Example 174: l-((5-((lR,3R)-2-(2-Fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro-lH- pyrido[3,4-b]indol-l-yl)-lH-indol-2-yl)methyl)-N,N-dimethylazetidin-3-amine
Figure imgf000193_0001
[0385] A solution of 5-((17?,37?)-2-(2-fluoro-2-methylpropyl)-3-methyl-2,3,4,9-tetrahydro- 12/-pyrido[3,4-Z>]indol-l-yl)-l/7-indole-2-carbaldehyde (60 mg, 0.15 mmol, 1 equiv) in di chloromethane (1.9 mL) was added to a mixture of N, /V-dim ethyl azeti din-3 -amine dihydrochloride salt (52 mg, 0.30 mmol, 2 equiv) and triethylamine (0.08 mL, 0.6 mmol, 4 equiv) in di chloromethane (1.9 mL). Acetic acid (0.02 mL, 0.3 mmol, 2 equiv) was added to the resulting reaction mixture which was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (0.095 g, 0.45 mmol, 3 equiv) was added and reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with dichloromethane (15 mL) and washed with saturated sodium carbonate (15 mL). The aqueous layer was extracted with di chloromethane (2 x 15 mL). The combined organic layers were dried over sodium sulfate, fdtered and concentrated under reduced pressure onto Celite® (1 g). The residue was purified on a Biotage automated chromatography system (5.5 g, Redigold C18 column), eluting with a gradient of 0 to 100% acetonitrile in water to give the title compound (50 mg, 69% yield) as an off-white solid. LCMS: m/z = 488.3 [M+H]+; XH NMR (400 MHz, CDCh) 5 = 8.60 (s, 1H), 7.83 (br s, 1H), 7.54 (d, J = 6.5 Hz, 1H), 7.33 - 7.26 (m, 1H), 7.23 - 7.07 (m, 5H), 6.21 (s, 1H), 5.06 (br s, 1H), 3.69 (s, 2H), 3.52 (br s, 1H), 3.48 - 3.37 (m, 2H), 2.98 - 2.79 (m, 4H), 2.75 - 2.53 (m, 3H), 2.08 (s, 6H), 1.43 - 1.22 (m, 6H), 1.06 (d, J= 6.7 Hz, 3H).
Example 175: (1R,3R)-1-(2-((3,3-Dimethylazetidin-1-yl)methyl)-1H-indol-5-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole [0386] A solution of 3,3 (37 mg, 0.297 mmol, 2 equiv)
Figure imgf000194_0001
and triethylamine (41 µL, 0.297 mmol, 2 equiv) in dichloromethane (2 mL) was added to a solution of 5-[(1R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-1,3,4,9-tetrahydropyrido[3,4- b]indol-1-yl]-1H-indole-2-carbaldehyde (60 mg, 0.149 mmol, 1 equiv) in dichloromethane (2 mL). Acetic acid (17 µL, 0.297 mmol, 2 equiv) was added and the resulting mixture was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (95 mg, 0.446 mmol, 3 equiv) was added in one portion, and the reaction was stirred at room temperature for 14 hours. The reaction was quenched with methanol (1 mL) and the volatiles were evaporated under reduced pressure. The residue was purified on an Interchim automated chromatography system (RediSep Rf GOLD 15 g HP C18 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound as an off-white solid. LCMS: m/z = 473.3 [M+H]+; 1H NMR (400 MHz, CDCl3) δ = 8.56 (br s, 1H), 7.63 - 7.47 (m, 2H), 7.27 - 7.07 (m, 7H), 6.20 (s, 1H), 5.04 (br s, 1H), 3.69 (s, 2H), 3.53 (br s, 1H), 2.99 (s, 4H), 2.93 - 2.78 (m, 1H), 2.76 - 2.52 (m, 3H), 2.35 (s, 1H), 1.40 (br s, 1H), 1.37 - 1.29 (m, 3H), 1.23 - 1.17 (m, 6H), 1.09 - 1.03 (m, 3H). Example 176: (1R,3R)-2-(2-Fluoro-2-methylpropyl)-1-(2-((3-methoxy-3-methylazetidin-1- yl)methyl)-1H-indol-5-yl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole
Figure imgf000194_0002
[0387] A solution of 3-methoxy-3-methylazetidine hydrochloride (41 mg, 0.297 mmol, 2 equiv) and triethylamine (41 µL, 0.297 mmol, 2 equiv) in dichloromethane (2 mL) was added to a solution of 5-[(1R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-1,3,4,9-tetrahydropyrido[3,4- b]indol-1-yl]-1H-indole-2-carbaldehyde (60 mg, 0.149 mmol, 1 equiv) in dichloromethane (2 mL). Acetic acid (17 µL, 0.297 mmol, 2 equiv) was added and the resulting mixture was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (95 mg, 0.446 mmol, 3 equiv) was added in one portion and the reaction was stirred at room temperature for 4 hours. The reaction was quenched with methanol (1 mL) and the volatiles were evaporated under reduced pressure. The residue was purified on an Interchim automated chromatography system (RediSep Rf GOLD 5.5 g HP C18 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound as a light yellow solid. LCMS: m/z = 489.3 [M+H]+; 1H NMR (400 MHz, CDCl3) δ = 8.41 (br s, 1H), 7.62 - 7.47 (m, 2H), 7.31 (s, 1H), 7.26 - 7.22 (m, 4H), 7.16 - 7.09 (m, 2H), 6.23 (s, 1H), 5.07 (br s, 1H), 3.77 (s, 2H), 3.55 (br s, 1H), 3.22 - 3.12 (m, 6H), 2.86 (br d, J = 13.2 Hz, 1H), 2.77 - 2.54 (m, 3H), 1.47 (s, 3H), 1.42 (br s, 1H), 1.39 - 1.24 (m, 5H), 1.08 (d, J = 6.6 Hz, 3H). Example 177: 1R,3R)-1-(2-((3,3-Difluoroazetidin-1-yl)methyl)-1H-indol-5-yl)-2-(2-fluoro-2- methylpropyl)-3-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole
Figure imgf000195_0001
9 mg, 0.297 mmol, 2 equiv) and triethylamine (41 µL, 0.297 mmol, 2 equiv) in dichloromethane (2 mL) was added to a solution of 5-[(1R,3R)-2-(2-fluoro-2-methyl-propyl)-3-methyl-1,3,4,9-tetrahydropyrido[3,4- b]indol-1-yl]-1H-indole-2-carbaldehyde (60 mg, 0.149 mmol, 1 equiv) in dichloromethane (2 mL). Acetic acid (17 µL, 0.297 mmol, 2 equiv) was added and the resulting mixture was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (143 mg, 0.669 mmol, 4.5 equiv) was added in one portion and the reaction was stirred at room temperature for 48 hours. The reaction was quenched with methanol (1 mL) and the volatiles were evaporated under reduced pressure. The residue was purified on an Interchim automated chromatography system (RediSep Rf GOLD 5.5 g HP Cl 8 column), eluting with a gradient of 0 to 100% acetonitrile in water to give the title compound as a yellow solid. LCMS: m/z = 481.3 [M+H]+; jH NMR (400 MHz, CDCh) 8 = 8.34 (br s, 1H), 7.61 - 7.49 (m, 2H), 7.34 - 7.27 (m, 2H), 7.23 (br d, J= 6.1 Hz, 1H), 7.18 - 7.10 (m, 2H), 6.27 (s, 1H), 5.10 (br s, 1H), 3.85 (s, 2H), 3.62 (t, J= 12.0 Hz, 4H), 3.57 - 3.47 (m, 1H), 2.85 (br d, J= 13.1 Hz, 1H), 2.78 - 2.52 (m, 3H), 1.52 - 1.26 (m, 7H), 1.08 (d, J = 6.6 Hz, 3H).
Example 178: (lR,3R)-2-(2-Fluoro-2-methylpropyl)-l-(2-((3-fluoro-3-methylazetidin-l- yl)methyl)-lH-indol-5-yl)-3-methyl-2,3,4,9-tetrahydro-lH-pyrido[3,4-b]indole
Figure imgf000196_0001
[0389] A solution of 3 -fluoro-3 -methylazetidine hydrochloride (37 mg, 0.297 mmol, 2 equiv) and tri ethylamine (41 pL, 0.297 mmol, 2 equiv) in dichloromethane (2 mL) was added to a solution of 5-[(17?,37?)-2-(2-fluoro-2-methyl-propyl)-3-methyl-l,3,4,9-tetrahydropyrido[3,4- Z>]indol-l-yl]-17/-indole-2-carbaldehyde (60 mg, 0.149 mmol, 1 equiv) in di chloromethane (2 mL). Acetic acid (17 pL, 0.297 mmol, 2 equiv) was added and the resulting mixture was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (143 mg, 0.669 mmol, 4.5 equiv) was added in one portion and the reaction was stirred at room temperature for 24 hours. The reaction was quenched with methanol (1 mb) and the volatiles were evaporated under reduced pressure. The residue was purified on an Interchim automated chromatography system (RediSep Rf GOLD 5.5 g HP Cl 8 column), eluting with a gradient of 0 to 100% acetonitrile in water, to give the title compound as a yellow solid. LCMS: m/z = 477.3 [M+H]+; 'H NMR (400 MHz, CDCh) 5 = 8.35 (br s, 1H), 7.62 - 7.48 (m, 2H), 7.32 (s, 1H), 7.26 - 7.20 (m, 3H), 7.17 - 7.09 (m, 2H), 6.25 (s, 1H), 5.08 (br s, 1H), 3.78 (s, 2H), 3.54 (br s, 1H), 3.39 - 3.27 (m, 4H), 2.86 (br d, J= 13.0 Hz, 1H), 2.78 - 2.55 (m, 3H), 1.64 - 1.55 (m, 3H), 1.46 - 1.25 (m, 6H), 1.08 (d, J = 6.7 Hz, 3H). Example 179: (5-((lR,3R)-3-Methyl-2-(2,2,2-trifluoroethyl)-2,3,4,9-tetrahydro-lH-pyrido[3,4- b]indol-l-yl)-lH-indol-2-yl)methanol
Figure imgf000197_0001
[0390] A mixture of 2-(hydroxymethyl)-l/f-indole-5-carbaldehyde (200 mg, 1.1 mmol, 1.0 equiv), (7?)-l-(l//-indol-3-yl)-A-(2,2,2-trifluoroethyl)propan-2-amine (290 mg, 1.1 mmol, 1.0 equiv), and glacial acetic acid (0.13 mL, 2.3 mmol, 2.0 equiv) in 1,2-di chloroethane (2.5 mL) and dioxane (2.5 mL) was stirred in a microwave apparatus (110 °C, 150 W, fixed power) for 6 h. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (20 mL) and washed with a saturated sodium carbonate solution (4 x 5 mL). The organic layer was dried over sodium sulfate (3 g), filtered and concentrated under reduced pressure. The crude product was diluted with di chloromethane (10 mL) and absorbed onto silica gel (5 g) under reduced pressure. The product was purified on a Biotage automated chromatography system (Biotage spherical silica gel column 25 g, 20 mm), eluting with a gradient of 0 to 10% methanol in dichloromethane to give the title compound (160 mg, 34% yield) as off-white solid. LCMS: m/z = 414 [M+H]“.
Example 180: 5-((lR,3R)-3-Methyl-2-(2,2,2-trifluoroethyl)-2,3,4, 9-tetrahydro-lH-pyrido[3,4- b]indol-l-yl)-lH-indole-2-carbaldehyde
Figure imgf000197_0002
[0391] A mixture of (5-((17^,3^)-3-methyl-2-(2,2,2-trifluoroethyl)-2,3,4,9-tetrahydro-lf7- pyrido[3,4-Z>]indol-l-yl)-l//-indol-2-yl)methanol (160 mg, 0.39 mmol, 1.0 equiv) and activated manganese dioxide (85%, 336 mg, 3.9 mmol, 10.0 equiv) in dichloromethane (3.9 mL) was stirred for 22 h. The reaction mixture was filtered through a pad of Celite® (10 g), which was washed with dichloromethane (3 x 10 mL). The filtrate was concentrated under reduced pressure. The crude product was diluted with dichloromethane (5 mL) and absorbed onto silica gel (2 g) under reduced pressure. The product was purified on a Biotage automated chromatography system (Biotage spherical silica gel column 5 g, 20 mm), eluting with a gradient of 0 to 50% ethyl acetate in hexanes to give the title compound (43 mg, 36% yield) as pale- yellow oil. LCMS: m/z = 412 [M+H]+. Example 181: (1R,3R)-1-(2-((3-(Fluoromethyl)azetidin-1-yl)methyl)-1H-indol-5-yl)-3-methyl- 2-(2,2,2-trifluoroethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole [0392] A mixture of
Figure imgf000198_0001
roethyl)-2,3,4,9-tetrahydro-1H- pyrido[3,4-b]indol-1-yl)-1H-indole-2-carbaldehyde (40 mg, 0.1 mmol, 1.0 equiv), 3- (fluoromethyl)azetidine hydrochloride (24 mg, 0.2 mmol, 2.0 equiv), 1.0 M triethylamine in dichloromethane (0.19 mL, 0.19 mmol, 2.0 equiv), and 1.0 M acetic acid in dichloromethane (0.19 mL, 0.19 mmol, 2.0 equiv) in dichloromethane (2 mL) was stirred for 1 hour. Sodium triacetoxyborohydride (62 mg, 0.3 mmol, 3.0 equiv) was added and the reaction mixture was stirred for 18 hours. Methanol (1 mL) was added and reaction mixture was stirred for 15 minutes. The mixture was concentrated under reduced pressure, diluted with dichloromethane (2 ml) and absorbed onto Celite® (1g) under reduced pressure. The product was purified on a Biotage automated chromatography system (RediSep Gold C18 column 5.5 g), eluting with a gradient of 0 to 80% acetonitrile in water. The product was lyophilized to give the title compound (23 mg, 49% yield) as white solid. 1H NMR (400 MHz, CDCl3) δ = 8.43 (br s, 1H), 7.65 - 7.44 (m, 2H), 7.32 - 7.27 (m, 2H), 7.26 - 7.05 (m, 4H), 6.27 - 6.21 (m, 1H), 5.05 (s, 1H), 4.57 (d, J = 5.7 Hz, 1H), 4.45 (d, J = 5.7 Hz, 1H), 3.72 (s, 2H), 3.51 - 3.33 (m, 3H), 3.24 (qd, J = 9.5, 15.4 Hz, 1H), 3.15 - 2.98 (m, 3H), 2.95 - 2.74 (m, 2H), 2.58 (dd, J = 8.1, 14.9 Hz, 1H), 1.14 (d, J = 6.7 Hz, 3H); LCMS: m/z = 485.2 [M+H]+. Example 182: 5-Bromo-2-(bromomethyl)-l,3-benzothiazole.
Figure imgf000199_0001
[0393] 5-Bromo-2-methylbenzothiazole (1.00 equiv, 2.00 g, 8.77 mmol), NBS (1.10 equiv, 1717 mg, 9.64 mmol), and AIBN (0.250 equiv, 360 mg, 2.19 mmol) were dissolved and stirred in carbon tetrachloride (20 mL) overnight at 90 °C. The reaction was then cooled to room temperature, and the insoluble material was filtered off. The solution was then dried under reduced pressure and purified via flash chromatography on silica gel in 0-20% EtOAc in hexanes to give the title product (590 mg, 22% yield). LCMS: m/z = 307.9 [M+H]+.
Example 183: 5-Bromo-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole.
Figure imgf000199_0002
[0394] 5 -Bromo-2-(bromomethyl)-l,3-benzothiazole (1.00 equiv, 590 mg, 1.92 mmol) and 3-(fluoromethyl)azetidine hydrochloride (1.20 equiv, 290 mg, 2.31 mmol) were dissolved in DMF (5 mL), and triethylamine (3.00 equiv, 0.80 mL, 5.77 mmol) was added. The solution was warmed to 50 °C and stirred for 1 h. The reaction was then cooled and poured into brine solution. The organic layers were separated and washed with saturated brine solution 5 times. They were then dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH in DCM to give the title product (473 mg, 78% yield). LCMS: m/z = 307.9 [M+H]+.
Example 184: 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole-5-carbonitrile.
Figure imgf000199_0003
[0395] 5 -Bromo-2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole (1.00 equiv,
454 mg, 1.44 mmol), tetrakis(triphenylphosphine)palladium(0) (0.0500 equiv, 83 mg, 0.0720 mmol), and zinc cyanide (1.00 equiv, 169 mg, 1.44 mmol) were dissolved in dry DMF (5 mL) under an atmosphere of nitrogen and stirred at 80 °C overnight. The solution was cooled, diluted with EtOAc, and poured into water. The organic layer was washed with brine 5 times, dried over sodium sulfate, fdtered, and dried in vacuo. The crude was purified on a 24 g silica column in 20-60% EtOAc in DCM to give the desired product (347 mg, 92.2% yield). LCMS: m/z = 262.3 [M+H]+.
Example 185: 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole-5-carbaldehyde.
Figure imgf000200_0001
[0396] 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole-5-carbonitrile (1.00 equiv, 347 mg, 1.33 mmol) was dissolved in dry THF (4 mL) under nitrogen and cooled to 0 °C. DIBAL (1 M in toluene, 1.46 mL) was added and stirred for 1 h. The reaction was quenched with EtOAc, then Rochelle's Salt solution was added and stirred for 30 min. The organic layers were separated, dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH (2% TEA) in DCM to give the desired product (170 mg, 48 yield). LCMS: m/z = 265.3 [M+H]+.
Example 186: 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2-methyl- propyl)-3-methyl-l, 3, 4, 9-tetrahydropyrido[3, 4-b]indol-l -yl]-l, 3-benzothiazole.
Figure imgf000200_0002
[0397] (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-l-amine (1.10 equiv, 176 mg, 0.707 mmol) and 2-[[3-(fluoromethyl)azetidin-l-yl]methyl]-l,3-benzothiazole-5- carbaldehyde (1.00 equiv, 170 mg, 0.643 mmol) were dissolved in DCE. Acetic acid (5.00 equiv, 0.18 mL, 3.22 mmol) was added, and the reaction mixture stirred at 110 °C under microwave irradiation for 2 h. The solution was cooled then diluted with DCM and poured into saturated sodium bicarbonate solution. The organic layers were separated, dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified on silica gel in 0-10% MeOH (2% TEA) in DCM to give the desired product at about 90% purity. 45 mg of this material was dissolved in DMF (2 mL) and purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) and lyophilized to give the title product (24.9 mg, 55% yield, >99% purity). LCMS: m/z = 495.7 [M+H]+; XH NMR (400 MHz, CDCh) 5 8.07 (s, 1H), 7.86 - 7.78 (m, 2H), 7.54 (dd, J = 20.7, 7.9 Hz, 2H), 7.17 (dt, J = 15.6, 7.1 Hz, 2H), 4.62 (d, J = 5.6 Hz, 1H), 4.50 (d, J = 5.6 Hz, 1H), 4.07 (s, 2H), 3.64 (t, J = 7.7 Hz, 2H), 3.34 (t, J = 7.0 Hz, 2H), 2.84 - 2.55 (m, 4H), 1.49 (d, J = 21.7 Hz, 2H), 1.37 (s, 2H), 1.30 (d, J = 11.9 Hz, 3H), 1.11 (d, J = 6.7 Hz, 3H).
Example 187: 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2-methyl-
Figure imgf000201_0001
[0398] 2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-5-[(lR,3R)-2-(2-fluoro-2-methyl -propyl)- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]-l,3-benzothiazole (33 mg, 0.07 mmol) and MCPBA (17 mg, 0.10 mmol) were dissolved in DCM and stirred at room temperature for 1 h. The reaction was poured into saturated sodium bicarbonate solution and extracted with DCM. The organic layers were separated and dried over sodium sulfate, fdtered, and concentrated in vacuo. The crude was dissolved in DMF (2 mL) and purified by HPLC on a Kintetex 5 pm Cl 8 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) and lyophilized to give the title product (11.9 mg, 35% yield). LCMS: m/z = 511.2 [M+H]+; XH NMR (400 MHz, CDCh) 8 7.90 - 7.81 (m, 2H), 7.66 (d, J = 9.0 Hz, 1H), 7.58 (d, J = 7.5 Hz, 1H), 7.33 (d, J = 7.8 Hz, 1H), 7.18 (dt, J = 18.9, 7.1 Hz, 2H), 5.29 (s, 1H), 5.04 (d, J = 15.6 Hz, 2H), 4.76 - 4.65 (m, 2H), 4.65 - 4.47 (m, 3H), 4.36 (dd, J = 18.0, 7.9 Hz, 1H), 2.85 - 2.54 (m, 4H), 1.47 (d, J = 19 6 Hz, 3H), 1.35 (d, J = 21.4 Hz, 3H), 1.28 (s, 1H), 1.11 (d, J = 6.7 Hz, 3H). Example 188: tert-Rutyl 3-(5-formylindazol-l-yl)azetidine-l -carboxylate and tert-butyl 3-(5- formylindazol-2-yl)azetidine-l-carboxylate
Figure imgf000202_0001
[0399] To a solution of lH-indazole-5-carbaldehyde (1.00 equiv, 797 mg, 5.45 mmol) in DMF (5 mL) in a water bath was added sodium hydride (1.50 equiv, 327 mg, 8.18 mmol). After 45 min, 2-methyl-2-propanyl 3-iodo-l-azetidinecarboxylate (1.30 equiv, 2.2 mL, 7.09 mmol) was added. The reaction was stirred for 15 min in a water bath before heating at 50 °C overnight. The reaction was quenched with water (50 mL) and extracted with EtOAc (50 mL). The organic layer was washed with 5% LiCl (50 mL x 2). The organic layers were dried over sodium sulfate and purified via silica gel (40 g) eluting with 25% hexanes:EtOAc to afford tert-butyl 3-(5-formylindazol-l-yl)azetidine-l-carboxylate as a beige solid (844 mg, 2.80 mmol, 51.4% yield) and with 35% hexanes:EtOAc to afford tert-butyl 3-(5-formylindazol-2- yl)azetidine-l -carboxylate as a beige solid (514 mg, 1.71 mmol, 31.3 % yield).
[0400] Nl : LCMS: m/z = 302.2 [M+H]+; ’ll NMR (400 MHz, DMSO) 5 10.05 (s, 1H), 8.53-8.45 (m, 2H), 7.91 (dd, J=8.8, 1.4Hz, 1H), 7.83 (d, J=8.9Hz, 1H), 5.77 (td, J=8.1, 4.0Hz, 1H), 4.41 (t, J=8.5Hz, 2H), 4.27 (s, 2H), 1.44 (s, 9H).
[0401] N2: LCMS: m/z = 302.1 [M+H]+; XH NMR (400 MHz, DMSO) 5 9.99 (d, J=0.8Hz,
1H), 8.89 (d, J=0.9Hz, 1H), 8.48 (dd, J=1.5, 0.9Hz, 1H), 7.89-7.75 (m, 1H), 7.71 (dd, J=9.0, 1.5Hz, 1H), 5.60 (tt, J=7.9, 5.1Hz, 1H), 4.43 (t, J=8.5Hz, 2H), 4.30 (s, 2H), 1.44 (s, 8H).
Example 189: tert-Butyl 3-[5-[(l R,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]-
Figure imgf000202_0002
[0402] A solution of (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)- 2,2-difluoropropan-l -amine (1.20 equiv, 456 mg, 0.900 mmol), tert-butyl 3-(5-formylindazol-l- yl)azetidine-l -carboxylate (1.00 equiv, 226 mg, 0.750 mmol), and acetic acid (3.00 equiv, 0.13 mL, 2.25 mmol) in DCE (1.5 mL) was heated at 125 °C for 3 h. DCM (20 mL) was added, and the organic layer was washed with water (10 mL) and brine (5 mL). The organic layers were dried over sodium sulfate and purified via silica gel (40 g) eluting with 60% hexanes:EtOAc to afford the title compound (579 mg, 0.733 mmol, 97.7 % yield) as a white solid. LCMS: m/z = 790.3 [M+H]+.
Example 190: tert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]-3-methyl-
Figure imgf000203_0001
[0403] To a solution of tert-butyl 3-[5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2- difluoro-propyl]-3 -methyl- 1 ,3 ,4,9-tetrahydropyrido[3,4-b]indol- 1 -yl]indazol- 1 -yl]azetidine- 1 - carboxylate (1.00 equiv, 468 mg, 0.592 mmol) in DCM (4.5 mL) at RT was added trifluoroacetic acid (11.0 equiv, 0.50 mL, 6.49 mmol). After 5 h, the reaction was azeotroped with DCE (5 mL x 2) to afford the title compound (639 mg, 0.696 mmol, 118 % yield) as a solid. LCMS: m/z = 690.4 [M+H]+.
Example 191: 2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]-3-methyl-l,3,4,9-
Figure imgf000203_0002
[0404] To a solution oftert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propoxy]-diphenyl-silane;2,2,2- trifluoroacetic acid (1.00 equiv, 52 mg, 0.0566 mmol) in THF (0.5 mb) was added tetra-n- butylammonium fluoride (5.30 equiv, 0.30 mL, 0.300 mmol). After 1 h, the reaction was left in at -28 °C overnight. The reaction was then stirred for 3 h at RT. The solvent was removed, NaHCCL (aq, 10 mL) was added and the reaction was extracted with DCM (10 mL x 2). The organic layers were dried over sodium sulfate, concentrated and purified via silica gel (12 g) eluting at 16% DCM/MeOH (2% NH4OH) to the title compound as a beige solid (179 mg, 0.396 mmol, 62.0 % yield). LCMS: m/z = 452.2 [M+H]+.
Example 192: 2,2-difluoro-3-[(lR,3R)-3-methyl-l-[l-(l-propylazetidin-3-yl)indazol-5-yl]-
Figure imgf000204_0001
[0405] A solution of 2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-l-ol (1.00 equiv, 30 mg, 0.0664 mmol), propanal (3.00 equiv, 0.015 mL, 0.199 mmol) and acetic acid (2.00 equiv, 0.0076 mL, 0.133 mmol) in methanol (1 mL) was stirred for 5 min before addition of sodium triacetoxyborohydride (4.00 equiv, 56 mg, 0.266 mmol). After stirring for 1 h, the reaction was directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm, gradient: 5-50% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a light yellow solid (11 mg, 0.0213 mmol, 32.0 % yield). LCMS: m/z = 494.4 [M+H]+;
Figure imgf000204_0002
NMR (400 MHz, DMSO) 8 10.83 (s, 1H), 8.28 (s, 1H), 8.06 (d, J = 0.8 Hz, 1H), 7.74 (d, J = 8.8 Hz, 1H), 7.52 - 7.44 (m, 2H), 7.33 - 7.25 (m, 2H), 7.08 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 7.00 (td, J = 7.5, 1.1 Hz, 1H), 5.39 (p, J = 7.1 Hz, 1H), 5.15 (s, 1H), 3.79 (dq, J = 9.7, 3.9 Hz, 3H), 3.45 (dt, J = 20.3, 7.2 Hz, 2H), 3.32 - 3.04 (m, 1H), 2.79 - 2.60 (m, 2H), 2.60 - 2.51 (m, 1H), 2.48 (d, J = 7.1 Hz, 2H), 1.36 (h, J = 7.3 Hz, 2H), 1.06 (d, J = 6.8 Hz, 3H), 0.89 (t, J = 7.4 Hz, 3H).
Example 193: 2,2-difluoro-3-[(lR,3R)-l-[l-[l-(3-fluoropropyl)azetidin-3-yl]indazol-5-yl]-3-
Figure imgf000205_0001
[0406] To a solution of 2,2-difluoro-3-[(lR,3R)-l-[l-(azetidin-3-yl)indazol-5-yl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-l-ol (1.00 equiv, 25 mg, 0.0554 mmol) and 1- bromo-3 -fluoropropane (1.00 equiv, 0.0051 mL, 0.0554 mmol) in DMF (1 mL) was added triethylamine (3.00 equiv, 0.023 mL, 0.166 mmol). After stirring overnight, more 1- bromo-3 -fluoropropane (0.2 equiv, 0.001 mL, 0.01 1 mmol) was added and the reaction heated at 50 °C. After 1 h, more l-bromo-3 -fluoropropane (0.2 equiv, 0.001 mL, 0.011 mmol) was added. After stirring for 1 h more at 50 °C, the reaction was directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-50% 0. 1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (5.8 mg, 0.0113 mmol, 20.5 % yield). LCMS: m/z = 512.4 [M+H]+; XH NMR (400 MHz, DMSO) 5 10.83 (s, 1H), 8.28 (s, 1H), 8.07(s, 1H), 7.74 (d, J=8.8Hz, 1H), 7.52-7.41 (m, 2H), 7.33-7.25(m, 2H), 7.12-7.04 (m, 1H), 7.04-6.94 (m, 1H), 5.40 (p, J=7.1Hz, 1H), 5.15 (s, 1H), 4.57 (t, J=6.0Hz, 1H), 4.45 (t, J=6.1Hz, 1H), 3.86-3.61 (m, 3H), 3.48 (dt, J=20.2, 7.1Hz, 2H), 3.29-3.08 (m, 1H), 1.73 (dp, J=25.7, 6 ,5Hz, 2H), 1.06 (d, J=6.8Hz, 3H).
Example 194: tert-butyl 3-[5-[(l R,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-difluoro-propyl]- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-l-yl]indazol-2-yl]azetidine-l-carboxylate
Figure imgf000206_0001
[0407] A solution of (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)- 2,2-difluoropropan-l -amine (1.50 equiv, 760 mg, 1.50 mmol), acetic acid (3.00 equiv, 0.17 mL, 3.00 mmol) and tert-butyl 3-(5-formylindazol-2-yl)azetidine-l-carboxylate (1.00 equiv, 301 mg, 1.00 mmol) in DCE (2 mL) was heated at 90 °C. After 2.5 h, the temperature was raised to 100 °C for 2 h. The reaction was further heated at 120 °C for 1.5 h. DCM (20 mL) was added, and the organic layers washed with water (10 mL) and brine (5 mL). The organic layers were dried over sodium sulfate and purified via silica gel (40 g) eluting with 60% hexanes :EtO Ac to afford the title compound as a clear, thick oil (998 mg, 1.26 mmol, 126 % yield). LCMS: m/z = 790.5 [M+H]+.
Example 195: tert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[2-(azetidin-3-yl)indazol-5-yl]-3-methyl-
Figure imgf000206_0002
[0408] To a solution of tert-butyl 3-[5-[(lR,3R)-2-[3-[tert-butyl(diphenyl)silyl]oxy-2,2- difluoro-propyl]-3 -methyl- 1,3,4, 9-tetrahydropyrido[3,4-b]indol-l-yl]indazol-2-yl]azeti dine-1- carboxylate (1.00 equiv, 358 mg, 0.453 mmol) in DCM (3.6 mL) was added trifluoroacetic acid (11.5 equiv, 0.40 mL, 5.19 mmol). After stirring overnight, sodium carbonate (aq, 5 mL) was added, and the reaction extracted with DCM (5 mL x 2) which was dried over sodium sulfate and concentated to afford the title compound as a white solid (280 mg, 0.406 mmol, 89.6 % yield).
LCMS: m/z = 690.7 [M+H]+.
Example 196: 2,2-difluoro-3-[(lR,3R)-l-[2-(azetidin-3-yl)indazol-5-yl]-3-methyl-l,3,4,9-
Figure imgf000207_0001
[0409] To a solution of tert-butyl-[2,2-difluoro-3-[(lR,3R)-l-[2-(azetidin-3-yl)indazol-5-yl]- 3-methyl-l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propoxy]-diphenyl-silane (1.00 equiv, 140 mg, 0.203 mmol) in THF (1 mL) was added tetra-n-butylammonium fluoride (1.97 equiv, 0.40 mL, 0.400 mmol) at room temperature. After 1 h, the reaction was concentrated and purified via silica gel (12 g) eluting at 16% DCM/MeOH (2% NFLOH) to afford the title compound as a white solid (52 mg, 0.115 mmol, 56.8 % yield). LCMS: m/z = 452.3 [M+H]+.
Example 197: 2,2-difluoro-3-[(lR,3R)-l-[2-(l-ethylazetidin-3-yl)indazol-5-yl]-3-methyl-
Figure imgf000207_0002
[0410] To a solution of 2,2-difluoro-3-[(lR,3R)-l-[2-(azetidin-3-yl)indazol-5-yl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-l-ol (1.00 equiv, 26 mg, 0.0576 mmol) in methanol (1 mL) was added acetaldehyde (3.00 equiv, 0.0098 mL, 0.173 mmol) and acetic acid (2.00 equiv, 0.0066 mL, 0.115 mmol). After 30 min, sodium triacetoxyborohydride (3.00 equiv, 37 mg, 0.173 mmol) was added. After stirring overnight, the reaction was directly purified by HPI.C on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm: gradient: 5-30% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (8.4 mg, 0.0161 mmol, 28.0 % yield). LCMS: m/z = 480.2 [M+H]+.
Example 198: 2,2-difluoro-3-[(lR,3R)-3-methyl-l-[2-(l-propylazetidin-3-yl)indazol-5-yl]~
Figure imgf000208_0001
[0411] To a solution of 2,2-difluoro-3-[(lR,3R)-l-[2-(azetidin-3-yl)indazol-5-yl]-3-methyl- l,3,4,9-tetrahydropyrido[3,4-b]indol-2-yl]propan-l-ol (1.00 equiv, 26 mg, 0.0576 mmol) in methanol (1 mL) was added propionaldehyde (3.00 equiv, 0.012 mL, 0.173 mmol) and acetic acid (2.00 equiv, 0.0066 mL, 0.115 mmol) . After 30 min, sodium triacetoxyborohydride (3.00 equiv, 37 mg, 0.173 mmol) was added. After stirring overnight, more propionaldehyde (3.00 equiv, 0.012 mL, 0.173 mmol) and sodium triacetoxyborohydride (3.00 equiv, 37 mg, 0.173 mmol) were added. After stirring overnight, the reaction was directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-30% 0.1% formic acid in ACN in 0,1% formic acid in water) then lyophilized to give the title compound as a white solid (8.1 mg, 0.0164 mmol, 28.5 % yield). LCMS: m/z = 494.4 [M+H]+; ’H NMR (400 MHz, DMSO) 5 10.83 (s, 1H), 8.38 (d, J = 1.0 Hz, 1H), 8.32 (s, 1H), 7.62 (d, J = 9.0 Hz, 1H), 7.50 - 7.36 (m, 2H), 7.34 - 7.26 (m, 1H), 7.12 (s, 1H), 7.07 (ddd, J = 8.1, 7.0, 1.3 Hz, 1H), 7.00 (td, J = 7.4, 1.1 Hz, 1H), 5.22 (p, J = 6.9 Hz, 1H), 5.09 (s, 1H), 3.77 (s, 1H), 3.74 (d, J = 7.7 Hz, 2H), 3.44 (td, J = 7.8, 6.4 Hz, 2H), 2 78 - 2.59 (m, 2H), 2.56 (d, J = 9.8 Hz, 1H), 2.46 (t, J = 7.1 Hz, 2H), 1.33 (h, J = 7.1 Hz, 2H), 1.06 (d, J = 6.9 Hz, 3H), 0.87 (t, J = 7.4 Hz, 3H). Example J 99: Ethyl 5-cyano-l-(p-tolylsulfonyl)indole-2-carboxylate
Figure imgf000209_0001
[0412] 5-Cyanoindole-2-carboxylic acid ethyl ester (1.00 equiv, 3.41 g, 15.9 mmol) was dissolved in DMF (25 mL) and cooled to 0 °C. Sodium hydride (1.24 equiv, 458 mg, 19.1 mmol) in mineral oil (60%) was added and stirred for 30 min. p-Toluenesulfonyl chloride (1.05 equiv, 3.19 g, 16.7 mmol) was added and stirred for 2 h while warming to room temperature. The reaction was quenched with IN HCL (40 mL) and extracted with EtOAc (2 x 50 mL). The combined organic fractions were washed with saturated brine solution (5 x 30 mL), dried over sodium sulfate, filtered, and dried in-vacuo. The crude was purified by flash chromatography in 0-22% EtOAc in hexanes to give the title product as a white solid (4.75 g, 12.89 mmol, 81% yield). LCMS: m/z = 369.1 [M+H]+.
Example 200: 2-(Hydroxymethyl)-l-(p-tolylsulfonyl)indole-5-carbaldehyde.
Figure imgf000209_0002
[0413] Ethyl 5 -cyano- l-(p-tolylsulfonyl)indole-2-carboxylate (1.00 equiv, 2.50 g, 6.79 mmol) was dissolved in anhydrous THF (34 mL) and cooled to 0 °C. DIBAL (IN in THF, 4.00 equiv, 27. 1 mL, 27. 1 mmol) was added slowly and stirred for 1 h. The reaction was diluted and quenched with EtOAc and saturated Rochelle's salt was added and stirred for Ih. The layers were separated and the aqueous phase was extracted with EtOAc (1 x 30 mL). The combined organic fractions were washed with saturated brine solution (1 x 30 mL), dried over sodium sulfate, filtered, and dried in vacuo. The crude was purified by flash chromatography in 0-25% EtOAc in hexanes to give the title product as an off-white solid. (1.1 g, 3.34 mmol, 49% yield). LCMS: m/z = 330.1 [M+H]+. Example 201: [5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(p-tolylsulfonyl)indol-2-yl]methanol
Figure imgf000210_0001
[0414] 2-(Hydroxymethyl)-l-(p-tolylsulfonyl)indole-5-carbaldehyde (1.00 equiv, 227 mg, 0.69 mmol) and 2,2,3,3-tetrafluoro-N-[(lR)-2-(lH-indol-3-yl)-l-methyl-ethyl]propan-l-amine (1.10 equiv, 0.219 mg, 0.76 mmol) were dissolved in dry toluene (2.5 mL) and acetic acid (4.9 equiv, 197 pL, 3.4 mmol) was added and stirred at 100 °C for 4h. The reaction was allowed to cool to room temperature and was quenched with saturated sodium bicarbonate solution and extracted with EtOAc (2 x 5 mL). The combined organic fractions were dried over sodium sulfate, filtered, and dried in-vacuo. The crude was purified by flash chromatography in 0-25% EtOAc in hexanes to give the title compound (256 mg, 0.43 mmol, 62% yield). LCMS: m/z = 600.1 [M+H]+.
Example 202: [5-[(lR,3R)-3-methyl-2-(2, 2,3,3, 3-pentafluoropropyl)-l, 3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-l-(p-tolylsulfonyl)indol-2-yl]methanol
Figure imgf000210_0002
[0415] 2-(Hydroxymethyl)-l-(p-tolylsulfonyl)indole-5-carbaldehyde (1.00 equiv, 239 mg, 0.73 mmol) and (R)-N-(l-(lH-indol-3-yl)propan-2-yl)-2,2,3,3,3-pentafluoropropan-l-amine (1.12 mmol, 0.250 mg, 0.82 mmol) were dissolved in dry toluene (2.5 mL) and acetic acid (4.9 equiv, 207 pL, 3.6 mmol) was added and stirred at 100 °C for 18h. The reaction was allowed to cool to room temperature and was quenched with saturated sodium bicarbonate solution and extracted with EtOAc (2 x 5 mL). The combined organic fractions were dried over sodium sulfate, filtered, and dried in-vacuo. The crude was purified by flash chromatography in 0-25% EtOAc in hexanes to give the title compound (261 mg, 0.42 mmol, 58% yield). LCMS: m/z = 618.1 [M+H]+.
Example 203: [5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9- tetrahydropyrido[3, 4-b]indol-l -yl]-lH-indol-2-yl]methanol
Figure imgf000211_0001
[0416] [5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(p-tolylsulfonyl)indol-2-yl]methanol (1.00 equiv, 256 mg, 0.43 mmol) and sodium hydroxide (31 equiv, 530 mg, 13.3 mmol) were stirred in MeOH (5 mL) for 3 h at 60 °C. The reaction was allowed to cool to room temperature and was recrystallized with water. The solids were collected with a filter funnel and washed with water (3 x 20 mL). This was dried under vacuum to give the title product (166 mg, 0.37 mmol, 87% yield). LCMS: m/z = 446.1 [M+H]+.
Example 204: [5-[(lR,3R)-3-Methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4, 9- tetrahydropyrido[3, 4-b]indol-l -yl]-LH-indol-2-yl] methanol
Figure imgf000211_0002
[0417] [5-[(lR,3R)-3-Methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-l-(p-tolylsulfonyl)indol-2-yl]methanol (1.00 equiv, 261 mg, 0.42 mmol) and sodium hydroxide (30 equiv, 507 mg, 12.7 mmol) were stirred in MeOH (1 mL) for 30 min at 60 °C. The reaction was allowed to cool to room temperature and was recrystallized with water. The solids were collected with a filter funnel and washed with water (3 x 20 mL). The collected solids were dried under vacuum to give the title product (142 mg, 0.31 mmol, 74% yield). LCMS: m/z = 464.1 [M+H]+.
Example 205: 5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde
Figure imgf000212_0001
[0418] [5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-2-yl]methanol (1.00 equiv, 166 mg, 0.37 mmol) and 45% IBX (3.00 equiv, 695 mg, 1.11 mmol) were stirred in DMSO (3.5 mL) and stirred for 1.5 h. The reaction was diluted with EtOAc and quenched with saturated sodium bicarbonate solution. This was extracted with EtOAc (10 mL) and the combined organic fractions were washed with saturated brine solution (3 x 10 mL). The organic phase was then dried over sodium sulfate, filtered, and dried in-vacuo. The crude was purified by flash chromatography in 0-30% EtOAc in hexanes to give the title compound (72 mg, 0.16 mmol, 44% yield). LCMS: m/z = 444.1 [M+H]+.
Example 206: 5-[(lR,3R)-3-Methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4,9- tetrahydropyrido[3,4-b]indol-l-yl]-lH-indole-2-carbaldehyde
Figure imgf000212_0002
[0419] [5-[(lR,3R)-3-Methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indol-2-yl]methanol (1.00 equiv, 142 mg, 0.31 mmol) and 45% IBX (3.00 equiv, 582 mg, 0.93 mmol) were stirred in DMSO (1.5 mL) and stirred for 1.5h. The reaction was diluted with EtOAc and quenched with saturated sodium bicarbonate solution. The reaction mixture was extracted with EtOAc (10 mL) and the combined organic fractions were washed with saturated brine solution (3 x 10 mL). The organic phase was then dried over sodium sulfate, fdtered, and dried in-vacuo. The crude was purified by flash chromatography in 0-40% EtOAc in hexanes to give the title compound (56 mg, 0.12 mmol, 42% yield). LCMS: m/z = 462.1 [M+H]+.
Example 207: (lR,3R)-l-[2-[[3-(Fluoromethyl)azetidin-l-yl]methyl]-lH-indol-5-yl]-3- methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4-b]indole
Figure imgf000213_0001
[0420] 5-[(lR,3R)-3-Methyl-2-(2,2,3,3-tetrafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (1.00 equiv, 72 mg, 0.16 mmol) was dissolved in DMF (1 mL) and acetic acid (2.00 equiv, 18 pL, 0.32 mmol) was added. Separately, 3- (fluoromethyl)azetidine hydrochloride (1.13 equiv, 36 mg, 0.18 mmol) was dissolved in DMF (1 mL) and TEA (2.2 equiv, 50 pL, 0.35 mmol) was added. These solutions were combined and stirred for 30 min. STAB (3.0 equiv, 104 mg, 0.48 mmol) was added and stirred for 1 h. The reaction was quenched with MeOH (1 mL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-45% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (36.7 mg, 0.07 mmol, 44% yield). LCMS: m/z = 517.1 [M+H]+ XH NMR (400 MHz, DMSO-d6) 5 10.95 (d, J = 2.2 Hz, 1H), 10.78 (s, 1H), 8.18 (s, 1H), 7.46 (d, J = 7.7 Hz, 1H), 7.27 (dd, J = 8.2, 4.5 Hz, 2H), 7.09 - 6.95 (m, 4H), 6.56 (ddd, J = 58.9, 52.4, 6.2 Hz, 1H), 6.17 (d, J = 1.8 Hz, 1H), 5.06 (s, 1H), 4.57 (d, J = 6.1 Hz, 1H), 4.45 (d, J = 6 1 Hz, 1H), 3.62 (s, 2H), 3.27 (q, J = 6.2 Hz, 5H), 2.97 (t, J = 6.7 Hz, 2H), 2.71 (ddd, J = 22.8, 14.5, 5.5 Hz, 2H), 2.60 - 2.53 (m, 1H), 1.05 (d, J = 6.7 Hz, 3H). Example 208: (1R,3R)-1 -[2-[[3- (Fluoromethyl) azetidin- 1 -yljmethyl]- 1 H-indol-5-yl]-3- methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4-b]indole
Figure imgf000214_0001
[0421] 5-[(lR,3R)-3-Methyl-2-(2,2,3,3,3-pentafluoropropyl)-l,3,4,9-tetrahydropyrido[3,4- b]indol-l-yl]-lH-indole-2-carbaldehyde (1.00 equiv, 56 mg, 0.12 mmol) was dissolved in DMF (1 mL) and acetic acid (2.00 equiv, 15 pL, 0.24 mmol) was added. Separately, 3- (fluoromethyl)azetidine hydrochloride (1.08 equiv, 17 mg, 0.13 mmol) was dissolved in DMF (1 mL) and TEA (2.2 equiv, 37 pL, 0.26 mmol) was added. These solutions were combined and stirred for 30 min. STAB (3.00 equiv, 77.2 mg, 0.36 mmol) was added and stirred for 1 h. The reaction was quenched with MeOH (1 mL) and directly purified by HPLC on a Kintetex 5 pm C18 100 A column (size: 100 x 30.0 mm; gradient: 5-45% 0.1% formic acid in ACN in 0.1% formic acid in water) then lyophilized to give the title compound as a white solid (29.8 mg, 0 056 mmol, 46% yield). LCMS: m/z = 535.1 XH NMR (400 MHz, DMSO-de) 5 10.97 (d, J = 2.1 Hz, 1H), 10.79 (s, 1H), 8.18 (s, 1H), 7.46 (d, J = 7.5 Hz, 1H), 7.27 (t, J = 8.1 Hz, 2H), 7.10 - 7.03 (m, 3H), 7.02 - 6.97 (m, 1H), 6.17 (d, J = 1.8 Hz, 1H), 5.07 (s, 1H), 4.57 (d, J = 6.1 Hz, 1H), 4.45 (d, J = 6.2 Hz, 1H), 3.62 (s, 2H), 3.54 (d, J = 16.8 Hz, 1H), 3.27 (qd, J = 7.6, 2.9 Hz, 2H), 2.97 (t, J = 6.6 Hz, 2H), 2.78 - 2.57 (m, 3H), 1.06 (d, J = 6.8 Hz, 3H).
Example 209: Biological Evaluation
[0422] ECC-1 cells were trypsinized and resuspended in hormone-depleted media and plated at a density of 15 k cells per well into a 96-well plate for at least 4 hours. Cells were treated with test compounds in the absence of E2 (for agonist mode) or in the presence of 500 pM E2 (for antagonist mode) for 3 days and plates were subsequently frozen at -80 °C. Thawed plates were incubated with a chromogenic substrate of AP, p-nitrophenyl phosphate (Thermo Fisher Scientific), for 40 minutes at 42 °C, and absorbances were read at 405 nm. AP activity was normalized to the activity of 500 pM E2 alone. This assay was shown to correlate with the in vivo studies comparing uterine wet weight in ovariectomized rats following treatment with a number of antiestrogens.
[0423] Table 6 shows estrogen receptor modulation (e.g., agonism and antagonism) of certain compounds of the present disclosure. The compound numbers correspond to the compound numbers of Table 1 and 2. Compounds having an activity designated as “+” provided estrogen receptor agonism having (i) at least 80% increase in the E2-normalized signal in the AP assay (agonist mode) and (ii) no more than 80% reduction in the E2-normalized signal in the AP assay (antagonist mode). Compounds having an activity designated as “++” provided estrogen receptor antagonism having (i) between 10% and 80% increase in the E2 -normalized signal in the AP assay (agonist mode) and (ii) between 10% and 80% reduction in the E2 -normalized signal in the AP assay (antagonist mode). Compounds having an activity designated as “+++” provided estrogen receptor antagonism (i) with pICso greater than 7.5 and at least a 10% reduction in the E2 -normalized signal in the AP assay (antagonist mode); and (ii) no more than 10% increase in E2 -normalized signal in the AP assay (agonist mode).
[0424] In some embodiments, particularly useful modulators of the estrogen receptor are compounds having greater than “+” activity in Table 6. In some embodiments, particularly useful modulators of the estrogen receptor are compounds having greater than “++” activity in Table 6. In some embodiments, particularly useful modulators of the estrogen receptor are compounds having “+++” activity in Table 6.
Table 6
Figure imgf000215_0001
Figure imgf000216_0001
Figure imgf000217_0001
Figure imgf000218_0001
[0425] Compound 1-109 and Compound 1-110 provided estrogen receptor antagonism with (i) pICso greater than 6.4 and at least a 10% reduction in the E2-normalized signal in the AP assay (antagonist mode); and (ii) no more than 10% increase in E2-normalized signal in the AP assay (agonist mode).
[0426] The embodiments of the disclosure described above are intended to be merely exemplary, numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention as defined in any appended claims.

Claims

CLAIMS A compound of F ormul a I :
Figure imgf000219_0001
T or a pharmaceutically acceptable salt thereof, wherein:
A is an optionally substituted 7- to 10-membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S or an optionally substituted 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S;
L is a covalent bond or an optionally substituted bivalent group selected from -Ci-Ce aliphatic-, -La-Co-C5 aliphatic-, and -C1-C5 aliphatic-La-, wherein La is selected from - S-, -SO-, -SO2-, and -N(Ra)-;
B is selected from -OH, -CO2H, C1-C6 aliphatic, 3- to 12-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S, and C3-C6 cycloaliphatic;
R1 is selected from hydrogen and optionally substituted Ci-Ce aliphatic;
R2 is selected from hydrogen and optionally substituted Ci-Ce aliphatic;
R3 is selected from hydrogen, halogen, -CN, -ORa, -C(O)Ra, -C(0)2Ra, -OC(O)Ra - C(0)N(Ra)2, -OC(O)N(Ra)2, -NO2, -N(Ra)2, -N(Ra)C(O)Ra, -N(Ra)C(O)2Ra, - N(Ra)S(O)2Ra, -SRa, -S(O)2Ra, -S(O)N(Ra)2, -S(O)2N(Ra)2, and an optionally substituted C1-6 aliphatic group; each R4 is independently oxo, halogen, -CN, -ORa, -N(Ra)2, -C(O)Ra, -OC(O)Ra, - C(O)2Ra, -C(O)N(Ra)2, -N(Ra)C(O)Ra, or an optionally substituted group selected from Ci-Ce aliphatic and 3- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from N, O, and S; each Ra is independently selected from hydrogen and optionally substituted Ci-Ce aliphatic; and n is 0 to 5. The compound of claim 1, wherein R1 is optionally substituted Ci-Ce aliphatic. The compound of claim 1 or 2, wherein R1 is Ci-Ce aliphatic optionally substituted with one or more halogen or -OH. The compound of claim 3, wherein R1 is Ci-Ce aliphatic optionally substituted with one or more fluoro or -OH.
The compound of any one of claims 1 -4, wherein R is
Figure imgf000220_0001
The compound of any one of claims 1-4, wherein R1 is
Figure imgf000220_0002
The compound of any one of claims 1-6, wherein R2 is optionally substituted Ci-Ce aliphatic. The compound of any one of claims 1-7, wherein R2 is methyl. The compound of any one of claims 1-8, wherein R3 is hydrogen. The compound of any one of claims 1-9, wherein A is an optionally substituted 7- to 10- membered bicyclic heteroaryl comprising 1-4 heteroatoms selected from N, O, and S.
11. The compound of any one of claims 1-10, wherein A is:
Figure imgf000221_0001
Figure imgf000222_0001
wherein Represents a point of attachment to moiety L. The compound of any one of claims 1-9, wherein A is an optionally substituted 3- to 6- membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. The compound of claim 12, wherein A is:
Figure imgf000222_0002
, wherein * represents a point of attachment to moiety L. The compound of any one of claims 1-13, wherein B is 3- to 6-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S. The compound of any one of claims 1-14, wherein B is 4- to 5-membered heterocyclyl comprising 1-3 heteroatoms selected from N, O, and S The compound of any one of claims 1-15, wherein B is azetidinyl or pyrrolidinyl. The compound of any one of claims 1-16, wherein B is azetidinyl. The compound of any one of claims 1-13, wherein B is 6- to 12-membered bicyclic fused or spirocyclic heterocyclyl. The compound of any one of claims 1-18, wherein:
Figure imgf000223_0001
is a moiety selected from:
Figure imgf000223_0002
The compound of any one of claims 1-19, wherein
Figure imgf000223_0003
Figure imgf000224_0001
The compound of any one of claims 1-13, wherein B is C3-C6 cycloaliphatic. The compound of claim 21, wherein B is selected from cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. The compound of any one of claims 1-22, wherein L is selected from optionally substituted Ci-Ce aliphatic, -S-C0-C5 aliphatic-, and -SO2-C0-C5 aliphatic. The compound of any one of claims 1-22, wherein L is optionally substituted Ci-Ce aliphatic. The compound of any one of claims 1-24, wherein L is -CH2-. The compound of any one of claims 1-19 or 21-25, wherein n is 1. The compound of any one of claims 1-19 or 21-25, wherein n is 0. The compound of any one of claims 1-26, wherein each R4 is independently selected from halogen and optionally substituted Ci-Ce aliphatic. The compound of any one of claims 1-26, wherein each R4 is an independently selected optionally substituted Ci-Ce aliphatic group. The compound of any one of claims 1-26, wherein each R4 is independently selected from fluoro, -CH3 and -CH2F. The compound of any one of claims 1-30, wherein the compound is of Formula II:
Figure imgf000225_0001
or a pharmaceutically acceptable salt thereof. The compound of claim 31, wherein the compound is of Formula Il-a:
Figure imgf000225_0002
or a pharmaceutically acceptable salt thereof. The compound of claim 31, wherein the compound is of Formula Il-b:
Figure imgf000226_0001
or a pharmaceutically acceptable salt thereof. The compound of claim 31, wherein the compound is of Formula II-c:
Figure imgf000226_0002
or a pharmaceutically acceptable salt thereof. The compound of claim 31, wherein the compound is of Formula Il-d:
Figure imgf000226_0003
TT-d or a pharmaceutically acceptable salt thereof.
36. The compound of claim 31, wherein the compound is of Formula Il-e:
Figure imgf000227_0001
or a pharmaceutically acceptable salt thereof.
37. The compound of claim 31, wherein the compound is of Formula Il-f:
Figure imgf000227_0002
or a pharmaceutically acceptable salt thereof.
38. The compound of any one of claims 1-11 or 14-31, wherein the compound is of Formula
III:
Figure imgf000228_0001
or a pharmaceutically acceptable salt thereof.
39. The compound of any one of claims 1-11 or 14-31, wherein the compound is of Formula
IV:
Figure imgf000228_0002
or a pharmaceutically acceptable salt thereof.
40. The compound of any one of claims 1-11 or 14-31, wherein the compound is of Formula
Figure imgf000228_0003
or a pharmaceutically acceptable salt thereof.
41. The compound of any one of claims 1-11 or 14-31, wherein the compound is of Formula
VI:
Figure imgf000229_0001
or a pharmaceutically acceptable salt thereof.
42. A compound selected from Table 1, or a pharmaceutically acceptable salt thereof.
43. A compound selected from Table 3, or a pharmaceutically acceptable salt thereof.
44. A compound selected from Table 4, or a pharmaceutically acceptable salt thereof.
45. A compound selected from Table 5, or a pharmaceutically acceptable salt thereof.
46. A pharmaceutical composition comprising a compound of any one of claims 1-45, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.
47. A method for treating a disorder mediated by an estrogen receptor in a subject, comprising administering to the subject a therapeutically effective amount of the compound of any one of claims 1-45, or a pharmaceutically acceptable salt thereof, or the composition of claim 46.
48. The method of claim 47, wherein the disorder is selected from the group consisting of breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, uterine cancer, and endometriosis. The method of claim 47 or 48, further comprising administering the compound or composition in combination or alternation with an anti-cancer agent. The method of claim 49, wherein the anti-cancer agent is selected from an mTOR inhibitor, a CDK4/6 inhibitor, a PI3 kinase inhibitor, an aromatase inhibitor, an antibody to or inhibitor of PD-1, PD-L1 or CTLA-4, or an antibody to or inhibitor of EGFR, PGFR, or IGFR. The method of claim 47 or 48, further comprising administering the compound or composition in combination or alternation with an estrogen receptor antagonist or a partial estrogen receptor antagonist. The method of any one of claims 47-51, wherein the disorder is breast cancer. A method of treating a subject suffering from a cancer comprising administering the compound of any one of claims 1-45, or a pharmaceutically acceptable salt thereof, or the composition of claim 46, in combination with an anti-cancer agent. The method of claim 53, wherein the anti-cancer agent is selected from an mTOR inhibitor, a CDK4/6 inhibitor, a PI3 kinase inhibitor, an aromatase inhibitor, an antibody to or inhibitor of PD-1, PD-L1 or CTLA-4, or an antibody to or inhibitor of EGFR, PGFR, or IGFR. The method of claim 54, wherein the anti-cancer agent is an mTOR inhibitor. The method of claim 55, wherein the mTOR inhibitor is selected from everolimus, sirolimus, temsirolimus, and LY3023414. The method of claim 54, wherein the anti-cancer agent is a CDK4/6 inhibitor. The method of claim 57, wherein the CDK4/6 inhibitor is selected from palbociclib, abemaciclib, ribociclib, lerociclib, trilaciclib, and SHR6390. The method of claim 54, wherein the anti-cancer agent is an antibody to or inhibitor of PD-1, PD-L1 or CTLA-4. The method of claim 54, wherein the anti-cancer agent is an antibody to or inhibitor of EGFR, PGFR, or IGFR. The method of claim 53, wherein the anti -cancer agent is a HER2 inhibitor. The method of claim 61, wherein the HER2 inhibitor is selected from tucatinib, trastuzumab, pertuzumab, ado-trastuzumab, trastuzumab emtansine, ado-trastuzumab emtansine, trastuzumab deruxtecan pertuzumab, lapatinib, and neratinib. The method of claim 54, wherein the anti -cancer agent is a PI3 kinase inhibitor. The method of claim 63, wherein the PI3 kinase inhibitor is selected from perifosine, CAL101, BEZ235, XL147, XL765, GDC-0941, and IPI-145. The method of claim 54, wherein the anti-cancer agent is a PIK3CA inhibitor. The method of claim 65, wherein the PIK3CA inhibitor is selected from alpelisib, taselisib, and LY3023414. The method of claim 54, wherein the anti-cancer agent is an aromatase inhibitor. The method of claim 67, wherein the aromatase inhibitor is selected from aminoglutethimide, testolactone, anastrozole, letrozole, exemestane, vorozole, formestane, fadrozole, 4-hydroxyandrostenedione, l,4,6-androstatrien-3, 17-dione, and 4- androstene-3,6, 17-trione. The method of claim 68, wherein the aromatase inhibitor is selected from anastrozole, letrozole, and exemestane. A method of preventing recurrence of a cancer in a subject comprising administering to the subject the compound of any one of claims 1-45, or a pharmaceutically acceptable salt thereof, or the composition of claim 46. The method of claim 70, wherein the cancer is selected from breast cancer, ovarian cancer, endometrial cancer, vaginal cancer, lung cancer, bone cancer, and uterine cancer. The method of claim 70 or 71, wherein the compound or composition is administered as an adjunctive therapy after or instead of chemotherapy, radiation, or surgery. The method of any one of claims 70-72, wherein the compound or composition is administered after surgery. The method of any one of claims 70-73, wherein the compound or composition is administered prior to surgery. The method of any one of claims 70-74, wherein the cancer is breast cancer. The method of claim 75, wherein the breast cancer has progressed in the presence of endocrine or aromatase therapy.
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