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WO2024148247A2 - Azaindazoles en tant qu'inhibiteurs de jak2 - Google Patents

Azaindazoles en tant qu'inhibiteurs de jak2 Download PDF

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Publication number
WO2024148247A2
WO2024148247A2 PCT/US2024/010456 US2024010456W WO2024148247A2 WO 2024148247 A2 WO2024148247 A2 WO 2024148247A2 US 2024010456 W US2024010456 W US 2024010456W WO 2024148247 A2 WO2024148247 A2 WO 2024148247A2
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compound
nitrogen
sulfur
oxygen
independently selected
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WO2024148247A3 (fr
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Craig E. Masse
Jiayi Xu
Kevin Robert DEMARCO
Phani GHANAKOTA
Jeremy R. Greenwood
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Ajax Therapeutics Inc
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Ajax Therapeutics Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • JAKs can interact with certain cytokine receptors and can couple cytokine binding to cytoplasmic signaling cascades, including the signal transducers and activators of transcription (STAT) pathway.
  • JAK proteins also contain a pseudokinase domain (JH2).
  • JH1 and JH2 contain an ATP binding site, but catalytic activity is believed to come predominantly from JH1, as JH2 lacks essential residues for phosphorylation catalysis.
  • JH2 mediates critical regulatory functions in JAKs and is believed to primarily serve to inhibit basal JAK2 activity.
  • provided compounds are useful for, among other things, treating and/or preventing diseases, disorders, or conditions associated with JAK2.
  • the present disclosure provides a compound of Formula I’: or a pharmaceutically acceptable salt thereof, wherein Ring A, Ring B, L 1 , L 2 , m, n, R 1 , R a , R b , X, and Y are as defined herein.
  • the present disclosure provides a compound of Formula I: or a pharmaceutically acceptable salt thereof, wherein Ring A, Ring B, m, n, R 1 , R a , R b , X, and Y are as defined herein.
  • Aliphatic refers to a straight-chain (i.e., unbranched) or branched, optionally substituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic 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 “carbocyclic” or “cycloaliphatic”), that has a single point 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., C 1-6 ).
  • Aryl refers to monocyclic and bicyclic ring systems having a total of six to fourteen ring members (e.g., C 6-14 ), wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members.
  • the term “aryl” may be used interchangeably with the term “aryl ring”.
  • “aryl” refers to an aromatic ring system which includes, but not limited to, phenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Unless otherwise specified, “aryl” groups are hydrocarbons.
  • heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridonyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[1,2- a]pyrimidinyl, imidazo[1,2-a]pyridinyl, thienopyrimidinyl, triazolopyridinyl, and benzoisoxazolyl.
  • 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 (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms).
  • Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H–quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3–b]–1,4–oxazin–3(4H)– one, and benzoisoxazolyl.
  • heteroaryl may be used interchangeably with the terms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any of which terms include rings that are optionally substituted.
  • Heteroatom refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic 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.
  • a bicyclic heterocyclic ring also includes groups in which the heterocyclic ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings.
  • Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3- dihydrobenzofuranyl, and tetrahydroquinolinyl.
  • Substituted applies to one or more hydrogens that are either explicit or implicit from the structure (e.g., refers to at least 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.
  • Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include –R ⁇ , –NR ⁇ 2 , –C(O)R ⁇ , –C(O)OR ⁇ , –C(O)C(O)R ⁇ , – C(O)CH 2 C(O)R ⁇ , -S(O) 2 R ⁇ , -S(O) 2 NR ⁇ 2 , –C(S)NR ⁇ 2 , –C(NH)NR ⁇ 2 , or –N(R ⁇ )S(O) 2 R ⁇ ; wherein each R ⁇ is independently hydrogen, C 1–6 aliphatic which may be substituted as defined below, 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 ⁇ ,
  • the present disclosure provides a compound of Formula I’-A: or a pharmaceutically acceptable salt thereof, wherein Ring A, Ring B, L 2 , m, n, R, R 1 , R a , and R b are as defined above for Formula I’ or Formula I and described in classes and subclasses herein, both singly and in combination, and x is 0, 1, 2, 3, or 4.
  • the present disclosure provides a compound of Formula I-B-1: or a pharmaceutically acceptable salt thereof, wherein Ring A, m, n, R 1 , R a , and R b are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • the present disclosure provides a compound of Formula I-C: or a pharmaceutically acceptable salt thereof, wherein Ring A, m, n, R 1 , R a , R b , X, and Y are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • the present disclosure provides a compound of Formula I-C-1: or a pharmaceutically acceptable salt thereof, wherein Ring A, m, n, R 1 , R a , and R b are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination.
  • the present disclosure provides a compound of Formula II: or a pharmaceutically acceptable salt thereof, wherein Ring A, Ring B, Cy 1 , n, p, R 1 , R a , R b , R c , X, and Y are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination; and m is 0, 1, 2, 3, or 4.
  • the present disclosure provides a compound of Formula II-C: or a pharmaceutically acceptable salt thereof, wherein Ring B, n, p, R 1 , R a , R b , R c , X, and Y are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination; and m is 0, 1, 2, 3, or 4.
  • the present disclosure provides a compound of Formula II-C: or a pharmaceutically acceptable salt thereof, wherein Ring B, n, p, R 1 , R a , R b , R c , X, and Y are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination; and m is 0, 1, 2, 3, or 4.
  • the present disclosure provides a compound of Formula II-D: or a pharmaceutically acceptable salt thereof, wherein Ring B, n, p, R 1 , R a , R b , R c , X, and Y are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination; and m is 0, 1, 2, 3, or 4.
  • the present disclosure provides a compound of Formula III-A: or a pharmaceutically acceptable salt thereof, wherein Ring A, Ring B, Cy 1 , Cy 2 , n, q, R 1 , R a , R b , R c , and R d are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination; m is 0, 1, 2, 3, or 4; and p is 0, 1, 2, 3, or 4.
  • the present disclosure provides a compound of Formula III-B- 1: or a pharmaceutically acceptable salt thereof, wherein Ring B, Cy 1 , Cy 2 , n, q, R 1 , R a , R b , R c , and R d are as defined above for Formula I and described in classes and subclasses herein, both singly and in combination; m is 0, 1, 2, 3, or 4; and p is 0, 1, 2, 3, or 4.
  • Ring A is absent such that in any Formulae described herein the place of Ring A is replaced with a hydrogen to satisfy valence and R a is absent.
  • L 1 is a bond and Ring A is absent such that in any Formulae described herein the entire side chain represented by is replaced by a hydrogen to satisfy valence.
  • the present disclosure provides a compound of or a pharmaceutically acceptable salt thereof, wherein Ring B, L 2 , n, R 1 , R b , X, and Y are as defined in classes and subclasses herein both singly and in combination.
  • L 2 is a bond.
  • L 3 is optionally substituted C 1 -C 6 aliphatic and one or more methylene groups are optionally and independently replaced with -C(O)N(R)- or -N(R)C(O)- and R is as described herein.
  • L 3 is , wherein z is 0, 1, 2, 3, 4, or 5.
  • L 3 is wherein z is 0, 1, 2, 3, 4, or 5 and * is a point of attachment to Ring A.
  • Y is CR y . In some embodiments, Y is CH. In some embodiments, Y is N. [0074] In some embodiments of any Formulae described herein, X is CR x and Y is CR y . In some embodiments, both X and Y are CH. [0075] In some embodiments of any Formulae described herein, R x is hydrogen, halogen, - CN, or optionally substituted C 1-6 alkyl. In some embodiments, R x is hydrogen. In some embodiments, R x is halogen (e.g., fluoro or chloro). In some embodiments, R x is –CN.
  • R x is optionally substituted C 1-6 aliphatic. In some embodiments, R x is optionally substituted C 1-6 alkyl.
  • R y is hydrogen, halogen, - CN, or optionally substituted C 1-6 alkyl. In some embodiments, R y is hydrogen. In some embodiments, R y is halogen (e.g., fluoro or chloro). In some embodiments, R y is –CN. In some embodiments, R y is optionally substituted C 1-6 aliphatic. In some embodiments, R y is optionally substituted C 1-6 alkyl.
  • Ring A is an optionally substituted group selected from phenyl, 5- to 6- membered monocyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 3- to 7-membered monocyclic cycloaliphatic, and 3- to 7-membered saturated or partially unsaturated monocyclic heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
  • Ring A is an optionally substituted group selected from 8- to 10-membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 4- to 10-membered bicyclic cycloaliphatic, and 4- to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
  • Ring A is an optionally substituted group selected from phenyl, 5- to 6-membered monocyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 8- to 10- membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
  • Ring A is 5- to 6-membered saturated or partially unsaturated monocyclic heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is 3- to 7-membered saturated monocyclic heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is 5- to 6-membered saturated monocyclic heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is a piperidine ring. [0084] In some embodiments, Ring A is optionally substituted 4- to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
  • Cy 1 is phenyl substituted with p R c groups.
  • Cy 1 is 5- to 6-membered monocyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and substituted with p R c groups.
  • Cy 1 is 5-membered monocyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and substituted with p R c groups.
  • Cy 1 is 5-membered monocyclic heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and substituted with p R c groups.
  • Cy 1 substituted with p R c groups is selected from: . [0108] In some embodiments, Cy 1 substituted with p R c groups is selected from: . [0109] In some embodiments, Cy 1 substituted with p R c groups is selected from: [0110] In some embodiments, Cy 1 substituted with p R c groups is selected from: .
  • the present disclosure provides methods of administering provided compounds or compositions to a subject in need thereof. In some embodiments, the present disclosure provides methods of administering provided compounds or compositions to a subject suffering from or susceptible to a disease, disorder, or condition associated with JAK2.
  • provided compounds are useful as JAK2 inhibitors. In some embodiments, provided compounds are useful as JAK2 inhibitors that bind the pseudokinase (JH2) domain of JAK2. In some embodiments, the present disclosure provides methods of inhibiting JAK2 in a subject comprising administering a provided compound or composition. In some embodiments, the present disclosure provides methods of inhibiting JAK2 in a biological sample comprising contacting the sample with a provided compound or composition.
  • JAK e.g., JAK2
  • JAK2 has been implicated in various diseases, disorders, and conditions, such as myeloproliferative neoplasms (Vainchenker, W. et al., F1000Research 2018, 7(F1000 Faculty Rev):82), atopic dermatitis (Rodrigues, M. A. and Torres, T. J. Derm. Treat. 2019, 31(1), 33-40) and acute respiratory syndrome, hyperinflammation, and/or cytokine storm syndrome (The Lancet. doi: 10.1016/80140-6736(20)30628-0).
  • a provided compound or composition is administered to a subject who is receiving or has received one or more additional therapies (e.g., an anti-cancer therapy and/or therapy to address one or more side effects of such anti-cancer therapy, or otherwise to provide palliative care).
  • additional therapies include BCL2 inhibitors (e.g., venetoclax), HDAC inhibitors (e.g., vorinostat), BET inhibitors (e.g., mivebresib), proteasome inhibitors (e.g., bortezomib), LSD1 inhibitors (e.g., IMG-7289), and CXCR2 inhibitors.
  • CXCR2 activity has been shown to modulate signaling pathways involved in tumor growth, angiogenesis, and/or metastasis, including the JAK-STAT3 pathway (Jaffer, T., Ma, D. Transl. Cancer Res.2016, 5(Suppl.4), S616-S628).
  • JAK-STAT3 pathway Jaffer, T., Ma, D. Transl. Cancer Res.2016, 5(Suppl.4), S616-S628).
  • EXAMPLES EXAMPLES [0150] As described in the Examples below, 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.
  • EA ethyl acetate
  • DMF ⁇ , ⁇ -dimethylformamide
  • Dess-Martin periodinane 1,1-dihydro-1,1,1-triacetoxy-l,2-benzoiodooxol-3(1H)-one
  • DCM Dichloromethane
  • THF Tetrahydrofuran
  • LAH Lithium aluminium hydride
  • DME 1,2-Dimethoxyethane
  • TEA Triethylamine
  • EIPEA Ethyldiisopropylamine
  • DEAD Diethylazodicarboxylate
  • DIAD Diisopropyl azodicarboxylate
  • HATU 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate
  • EDCI N-(3-dimethylamino
  • Example I-3 (R)-N-(1-methyl-3-(1-(tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)-1H-pyrazolo[3,4- c]pyridin-5-yl)cyclopropanecarboxamide [0159] Synthesis of compound 3.1. To a solution of 5-bromo-3-iodo-1H-pyrazolo[3,4- c]pyridine (0.2 g, 0.617 mmol, 1 equiv) in DMF (4 mL) was added sodium hydride (0.054 g, 1.234 mmol, 2 equiv) at 0 oC and stirred at room temperature of 15 min.
  • Compound 3.2 was prepared from compound 3.1 and 2.2, following the procedure described in the synthesis of compound 2.4. The product was purified by flash column chromatography on silica gel (Combiflash®, 45% ethyl acetate in hexane). MS(ES): m/z 349.2 [M+H] + . [0161] Synthesis of I-3. Compound I-3 was prepared from compound 3.2, following the procedure described in the synthesis of compound 2.5. The product was purified by flash column chromatography on silica gel (Combiflash®, 80% ethyl acetate in hexane).
  • Example I-6 N-(3-(3-(1-cyclobutyl-1H-pyrazol-4-yl)-2-methoxyphenyl)-1-methyl-1H- pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide [0170] Synthesis of compound 6.1. To a solution of 4-bromo-1H-pyrazole (5.0 g, 34.013 mmol, 1.0 equiv) in DMF (50 mL) was added sodium hydride (60 % in mineral oil, 3.4 g, 85.034 mmol, 2.5 equiv) in portions at 0 °C and stirred for 30 min.
  • N, N-dimethylethylenediamine (0.253 g, 2.87 mmol, 0.3 equiv) and copper iodide (0.366 g, 1.91 mmol, 0.2 equiv) were added, and degassed for 5 min.
  • the reaction mixture was stirred at 120 oC for 16 h. It was cooled to room temperature, filtered through a pad of Celite®. The filtrate was transferred into water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sufhate, filtered, and concentrated under reduced pressure.
  • Example I-10 N-(3-(2-methoxy-3-(1-((1S,2S)-2-methoxycyclopentyl)-1H-pyrazol-4-yl)phenyl)- 1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide and N-(3-(2-methoxy-3-(1- ((1R,2R)-2-methoxycyclopentyl)-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin- 5-yl)cyclopropanecarboxamide [0195] Synthesis of compound trans-( ⁇ )-10.1.
  • Example I-17 (R)-N-(3-(3-(1-(tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)phenyl)-1H-pyrazolo[3,4- c]pyridin-5-yl)cyclopropanecarboxamide
  • Compound 17.1 was prepared from compound 15.3 and 2.3, following the procedure described in the synthesis of compound ( ⁇ )-9.5. The product his was purified by flash column chromatography on silica gel (Combiflash®, 32% ethyl acetate in hexane). MS(ES): m/z 541.37 [M+H] + .
  • Compound I-27 was prepared from compound 27.6 and cyclopropanecarboxamide, following the procedure described in the synthesis of compound 2.5.
  • the product was purified by flash column chromatography on silica gel (Combiflash®, 2.2% methanol in DCM).
  • Tris(dibenzylideneacetone)dipalladium(0) (0.065 g, 0.08 mmol, 0.1 equiv) was added and degassed for 5 min.
  • the reaction mixture was stirred at 100 oC for 1.5 h. It was cooled to room temperature and filtered through a pad of Celite®. The filtrate was transferred into water, extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (Combiflash®, 43% ethyl acetate in hexane) to afford ( ⁇ )-29.6 (0.25 g, 83%).
  • Example I-31-a and I-31-b 6-(cyclopropanecarboxamido)-4-((3-methoxy-5-(1-((3R,4S)-4- methoxytetrahydrofuran-3-yl)-1H-pyrazol-4-yl)pyridin-2-yl)amino)nicotinamide and 6- (cyclopropanecarboxamido)-4-((3-methoxy-5-(1-((3R,4S)-4-methoxytetrahydrofuran-3-yl)-1H- pyrazol-4-yl)pyridin-2-yl)amino)nicotinamide [0319] Synthesis of compound 31.1.
  • Example I-37-a and I-37-b (R)-N-(3-(2-methoxy-3-(1-(2-oxopiperidin-3-yl)-1H-pyrazol-4- yl)phenyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide and (S)-N-(3-(2- methoxy-3-(1-(2-oxopiperidin-3-yl)-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazolo[3,4- c]pyridin-5-yl)cyclopropanecarboxamide [0357] Synthesis of compound ( ⁇ )-37.1.
  • the racemate was separated by chiral SFC (column CHIRALPAK IC (250 x 21 mm, 5 ⁇ m); mobile phase: (A) Liquid CO 2 , (B) 0.1% diethylamine in propane-2-ol: acetonitrile (50: 50); flow rate: 80 mL/min) to afford first eluting fraction (I-38-a) and second eluting fraction (I-38-b). *The absolute configuration of the chiral center is not determined.
  • Example I-40 methyl 4-cyano-4-(3-(5-(cyclopropanecarboxamido)-1-methyl-1H-pyrazolo[3,4- c]pyridin-3-yl)-2-methoxyphenyl)piperidine-1-carboxylate [0386] Synthesis of compound 40.1. To a solution of 35.2 (0.45 g, 1.52 mmol, 1.0 equiv) and triethylamine (0.463 g, 4.57 mmol, 3.0 equiv) in DCM (5.0 mL) at 0 °C was added dropwise acetyl chloride (0.173 g, 1.83 mmol, 1.2 equiv) and stirred for 2 h.
  • Compound I-40 was prepared from compound 40.3 and cyclopropanecarboxamide, following the procedure described in the synthesis of compound 2.5.
  • the product was purified by flash column chromatography on silica gel (Combiflash®, 3.1% methanol in DCM).
  • Example I-41 4-cyano-4-(3-(5-(cyclopropanecarboxamido)-1-methyl-1H-pyrazolo[3,4- c]pyridin-3-yl)-2-methoxyphenyl)-N,N-dimethylpiperidine-1-carboxamide [0390] Synthesis of compound 41.1. To a solution of 35.2 (0.45 g, 1.52 mmol, 1.0 equiv) and triethylamine (0.463 g, 4.57 mmol, 3.0 equiv) in DCM (5.0 mL) at 0 °C was added dimethylcarbamic chloride (0.197 g, 1.83 mmol, 1.2 equiv) and stirred for 2 h.
  • Compound I-41 was prepared from compound 41.3 and cyclopropanecarboxamide, following the procedure described in the synthesis of compound 2.5.
  • the product was purified by flash column chromatography on silica gel (Combiflash®, 3.2% methanol in DCM).
  • Example I-42 N-(3-(3-(4-cyano-1-(2-methoxyethyl)piperidin-4-yl)-2-methoxyphenyl)-1- methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide [0394] Synthesis of compound 42.1. To a solution of 35.2 (0.450 g, 1.52 mmol, 1.0 equiv) and triethylamine (0.63 mL, 4.57 mmol, 3.0 equiv) in DCM (10 mL) was added 1-bromo-2- methoxyethane (0.423 g, 3.05 mmol, 2.0 equiv) at 0 °C.
  • Example I-45-a and I-45-b (S)-N-(3-(2-methoxy-3-(1-((3-methoxytetrahydrofuran-3- yl)methyl)-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5- yl)cyclopropanecarboxamide and (R)-N-(3-(2-methoxy-3-(1-((3-methoxytetrahydrofuran-3- yl)methyl)-1H-pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5- yl)cyclopropanecarboxamide [0409] Synthesis of compound ( ⁇ )-45.1.
  • Example I-47-a and I-47-b N-(3-(3-(1-((5R,9S)-1,7-dioxaspiro[4.4]nonan-9-yl)-1H-pyrazol-4- yl)-2-methoxyphenyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide and N-(3-(3-(1-((5S,9R)-1,7-dioxaspiro[4.4]nonan-9-yl)-1H-pyrazol-4-yl)-2-methoxyphenyl)-1- methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide [0427] Synthesis of compound ( ⁇ )-47.1.
  • Example I-48 N-(3-(5-methoxy-1-(2-oxaspiro[3.3]heptan-6-yl)-1H-pyrazol-4-yl)-1-methyl-1H- pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide [0437] Synthesis of compound 48.1.
  • Compound I-48 was prepared from compound 48.5 and cyclopropanecarboxamide, following the procedure described in the synthesis of compound 2.5.
  • the product was purified by flash column chromatography on silica gel (Combiflash®, 5% methanol in DCM).
  • Example I-49 N-(3-(3-(1-(ethylsulfonyl)-4-(hydroxymethyl)piperidin-4-yl)-2-methoxyphenyl)- 1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide [0442] Synthesis of compound 49.1. To a stirred solution of I-35 (0.20 g, 0.383 mmol, 1.0 equiv) in THF at 0 °C was added diisobutylaluminum hydride (0.547 mL, 1.14 mmol, 3.0 equiv) dropwise and stirred at room temperature for 5 h.
  • diisobutylaluminum hydride 0.547 mL, 1.14 mmol, 3.0 equiv
  • Example I-50-a and I-50-b (S)-N-(3-(5-methoxy-1-(tetrahydro-2H-pyran-3-yl)-1H-pyrazol-4- yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide and (R)-N-(3-(5- methoxy-1-(tetrahydro-2H-pyran-3-yl)-1H-pyrazol-4-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5- yl)cyclopropanecarboxamide [0444] Synthesis of compound ( ⁇ )-50.1.
  • the racemate was separated by chiral HPLC (column: CHIRALPAK IB-N (250 x 21 mm, 5 ⁇ m); mobile phase: (A) 0.1% diethylamine in n-hexane, (B) 0.1% diethylamine in propane-2-ol: methanol (50: 50); flow rate: 17 mL/min) to afford first eluting fraction (I-50-a) and second eluting fraction (I-50-b).
  • chiral HPLC column: CHIRALPAK IB-N (250 x 21 mm, 5 ⁇ m); mobile phase: (A) 0.1% diethylamine in n-hexane, (B) 0.1% diethylamine in propane-2-ol: methanol (50: 50); flow rate: 17 mL/min) to afford first eluting fraction (I-50-a) and second eluting fraction (I-50-b).
  • the absolute configuration of the chiral center is not determined.
  • Compound 51.6-a was prepared from compound 51.5- a, following the procedure described in the synthesis of compound ( ⁇ )-9.4. The product was purified by flash column chromatography on silica gel (Combiflash®, 35% ethyl acetate in hexane). MS(ES): m/z 387.3 [M+H] + . [0459] Synthesis of compound 51.6-b. Compound 51.6-b was prepared from compound 51.5- b, following the procedure described in the synthesis of compound ( ⁇ )-9.4. The product was purified by flash column chromatography on silica gel (Combiflash®, 35% ethyl acetate in hexane).
  • Compound I-52 was prepared from compound 52.5 and cyclopropanecarboxamide, following the procedure described in the synthesis of compound 2.5.
  • the product was purified by flash column chromatography on silica gel (Combiflash®, 2.2% methanol in DCM).
  • Example I-53 N-(3-(3-(1-((1r,3r)-3-fluorocyclobutyl)-1H-pyrazol-4-yl)-2-methoxyphenyl)-1- methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide [0469] Synthesis of compound cis-53.1. To a solution of 3-(benzyloxy)cyclobutan-1-one (25 g, 142.05 mmol, 1.0 equiv) in methanol (250 mL), was added sodium borohydride (16.19 g, 426.13 mmol, 2.0 equiv) in portions at 0 °C.
  • Compound ( ⁇ )-55.4 was prepared from compound ( ⁇ )-55.3 and 2-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline, following the procedure described in the synthesis of compound ( ⁇ )-11.3.
  • the product material was purified by flash column chromatography on silica gel (Combiflash®, 50% ethyl acetate in hexane). MS(ES): m/z 302.22 [M+H] + .
  • Synthesis of compound ( ⁇ )-55.5 Compound ( ⁇ )-55.5 was prepared from compound ( ⁇ )-55.4, following the procedure described in the synthesis of compound ( ⁇ )-9.4.
  • Example I-57 N-(3-(2-methoxy-3-(1-((3R,6R)-6-(methoxymethyl)-2-oxopiperidin-3-yl)-1H- pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide
  • Trimethylbenzeniminium dibromide (4.70 g) was added in portions. The reaction mixture was stirred at room temperature for 8 h. It was transferred into water and extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (Combiflash®, 50% ethyl acetate in hexane) to afford 57.5 (4.1 g, 60%). MS(ES): m/z 312.3 and 314.3[M+H] + . [0508] Synthesis of compound 57.6.
  • Example I-58-a and I-58-b (R)-N-(3-(2-methoxy-3-(1-(5-oxo-4-azaspiro[2.5]octan-6-yl)-1H- pyrazol-4-yl)phenyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide and (S)-N-(3-(2-methoxy-3-(1-(5-oxo-4-azaspiro[2.5]octan-6-yl)-1H-pyrazol-4-yl)phenyl)-1- methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide
  • the racemate was separated by chiral SFC (column: CHIRALPAK IC (250 x 21 mm, 5 ⁇ m); mobile phase: (A) CO 2 , (B) 0.1% diethylamine in isopropanol: MeCN (50: 50); flow rate: 80 mL/min) to afford first eluting fraction (I-58-a) and second eluting fraction (I-58-b). *The absolute configuration of the chiral center is not determined.
  • the racemate was separated by chiral SFC (column: CHIRALPAK IB-N (250 x 21 mm, 5 ⁇ m); mobile phase: (A) CO 2 , (B) 0.1% diethylamine IN isopropanol: MeCN (50: 50); flow rate: 80 mL/min) to afford first eluting fraction (I-59-a) and second eluting fraction (I-59-b). *The absolute configuration of the chiral center is not determined.
  • Example I-64-a and I-64-b N-(3-(4-(1-((5R,9S)-1,7-dioxaspiro[4.4]nonan-9-yl)-1H-pyrazol-4- yl)-3-methoxypyridin-2-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide and N-(3-(4-(1-((5S,9R)-1,7-dioxaspiro[4.4]nonan-9-yl)-1H-pyrazol-4-yl)-3-methoxypyridin-2- yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)cyclopropanecarboxamide [0573] Synthesis of compound ( ⁇ )-64.1.
  • the remaining kinases are produced in HEK-293 cells and subsequently tagged with DNA for qPCR detection.
  • Streptavidin-coated magnetic beads are treated with biotinylated small molecule ligands for 30 minutes at room temperature to generate affinity resins for kinase assays.
  • the liganded beads are blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific binding.
  • blocking buffer SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT
  • Binding reactions are assembled by combining kinases, liganded affinity beads, and test compounds in 1x binding buffer (20% SeaBlock, 0.17x PBS, 0.05% Tween 20, 6 mM DTT).
  • Test compounds are prepared as 111X stocks in 100% DMSO. Kds are determined using an 11-point 3-fold compound dilution series with three DMSO control points. All compounds for Kd measurements are distributed by acoustic transfer (non- contact dispensing) in 100% DMSO. The compounds are then diluted directly into the assays such that the final concentration of DMSO is 0.9%. All reactions are performed in polypropylene 384- well plate. Each has a final volume of 0.02 ml.
  • the cells are spun, and the cell pellets are re-suspended with 100 ⁇ L cold PBS. Then, the cells are spun again at 4 °C and 4000 rpm for 5 min. PBS is aspirated, and 25 ⁇ L lysis buffer (with protease and phosphatase inhibitor cocktail) is added to each cell pellet. The cell lysate is shaken at 4 °C for 20 min to fully lyse the cells. The cell lysate is spun at 4 °C and 4000 rpm for 15 min, and then the supernatant is transferred into a new plate and stored at -80 °C.
  • lysis buffer with protease and phosphatase inhibitor cocktail
  • MSD Meso-scale discovery
  • PBS 40 ⁇ L/well
  • 1x MSD Wash Buffer Tris-buffered saline with 0.1% Tween® 20 detergent, TBST
  • the MSD plates are then blocked with 150 ⁇ L of blocking buffer (5% BSA in TBST) and shaken for 1 h at room temperature and 600 rpm.
  • the MSD plate is washed three times with 150 ⁇ L/well of 1x MSD Wash Buffer (TBST).
  • Sample lysates are then added to MSD plates (25 ⁇ L/well) and shaken for 1 h at room temperature and 600 rpm.
  • the MSD plate is washed three times with 150 ⁇ L/well of 1x MSD Wash Buffer (TBST).
  • Detection antibody prepared in Antibody Detection buffer, 1% BSA in 1xTBST is then added to the MSD plates, and they are shaken for 1 h at room temperature and 600 rpm.
  • the MSD plate is washed three times with 150 ⁇ L/well of 1x MSD Wash Buffer (TBST).
  • Assay Procedure The Caco-2 plate is removed from the incubator and washed twice with pre-warmed HBSS (10 mM HEPES, pH 7.4), and then incubated at 37 °C for 30 minutes.
  • the stock solutions of control compounds are diluted in DMSO to get 1 mM solutions and then diluted with HBSS (10 mM HEPES, pH 7.4) to get 5 ⁇ M working solutions.
  • the stock solutions of the test compounds are diluted in DMSO to get 1 mM solutions and then diluted with HBSS (10 mM HEPES and 4% BSA, pH 7.4) to get 5 ⁇ M working solutions.
  • the final concentration of DMSO in the incubation system is 0.5%.
  • Time 0 samples are prepared by transferring 50 ⁇ L of 5 ⁇ M working solution to wells of the 96-deepwell plate, followed by the addition of 200 ⁇ L cold methanol containing appropriate internal standards (IS). The plates are incuabted at 37 °C for 2 hours. At the end of the incubation, 50 ⁇ L samples from donor sides (apical compartment for Ap ⁇ Bl flux, and basolateral compartment for Bl ⁇ Ap) and receiver sides (basolateral compartment for Ap ⁇ Bl flux, and apical compartment for Bl ⁇ Ap) are transferred to wells of a new 96-well plate, followed by the addition of 4 volume of cold acetonitrile or methanol containing appropriate internal standards (IS).
  • Lucifer Yellow leakage after 2 hour transport period stock solution of Lucifer yellow is prepared in ultra-pure water and diluted with HBSS (10 mM HEPES, pH 7.4) to reach the final concentration of 100 ⁇ M.100 ⁇ L of the Lucifer yellow solution is added to each Transwell® insert (apical compartment), followed by filling the wells in the receiver plate (basolateral compartment) with 300 ⁇ L of HBSS (10 mM HEPES, pH 7.4).
  • Hepatocyte Stability Assay 10 mM stock solutions of test compound and positive control are prepared in DMSO. Stock solutions are diluted to 100 ⁇ M by combining 198 ⁇ L of 50% acetonitrile/50% water and 2 ⁇ L of 10 mM stock solution. Verapamil is used as positive control in the assay. Vials of cryopreserved hepatocytes are thawed in a 37 °C water bath with gently shaking. The contents are poured into the 50 mL thawing medium conical tube. Vials are centrifuged at 100 g for 10 minutes at room temperature.
  • Thawing medium is aspirated and hepatocytes are re-suspended with serum- free incubation medium to yield ⁇ 1.5 ⁇ 106 cells/mL.
  • Cell viability and density are counted using a Trypan Blue exclusion, and then cells are diluted with serum-free incubation medium to a working cell density of 0.5 ⁇ 106 viable cells/mL.
  • a portion of the hepatocytes at 0.5 ⁇ 106 viable cells/mL are boiled for 5 min prior to adding to the plate as negative control to eliminate the enzymatic activity so that little or no substrate turnover should be observed.
  • Aliquots of 198 ⁇ L hepatocytes are dispensed into each well of a 96-well non-coated plate. The plate is placed in the incubator for approximately 10 minutes.
  • test compounds are calculated as follows, wherein INJ VOL is injection volume, DF is dilution factor, and STD is standard: Plasma Protein Binding Assay [0591]
  • Working solutions of test compounds and control compound are prepared in DMSO at the concentration of 200 ⁇ M, and then the working solutions are spiked into plasma. The final concentration of compound is 1 ⁇ M. The final concentration of DMSO is 0.5%.
  • Ketoconazole is used as positive control in the assay.
  • Dialysis membranes are soaked in ultrapure water for 60 minutes to separate strips, then in 20% ethanol for 20 minutes, finally in dialysis buffer for 20 minutes. The dialysis set up is assembled according to the manufacturer’s instruction.

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  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

La présente invention concerne des composés azaindazole et des compositions de ceux-ci utiles pour inhiber JAK2.
PCT/US2024/010456 2023-01-06 2024-01-05 Azaindazoles en tant qu'inhibiteurs de jak2 Ceased WO2024148247A2 (fr)

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EP2742046A1 (fr) * 2011-08-12 2014-06-18 F.Hoffmann-La Roche Ag Composés de pyrazolo[3,4-c]pyridine et procédés d'utilisation
US10155987B2 (en) * 2012-06-12 2018-12-18 Dana-Farber Cancer Institute, Inc. Methods of predicting resistance to JAK inhibitor therapy
WO2016026549A1 (fr) * 2014-08-22 2016-02-25 Merck Patent Gmbh Indazoles
MA46191A (fr) * 2016-09-09 2021-04-21 Incyte Corp Dérivés de pyrazolopyridine comme modulateurs de hpk1 et leurs utilisations pour le traitement du cancer
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