WO2021175271A1 - 新型hpk1抑制剂及其制备方法和应用 - Google Patents

新型hpk1抑制剂及其制备方法和应用 Download PDF

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WO2021175271A1
WO2021175271A1 PCT/CN2021/078966 CN2021078966W WO2021175271A1 WO 2021175271 A1 WO2021175271 A1 WO 2021175271A1 CN 2021078966 W CN2021078966 W CN 2021078966W WO 2021175271 A1 WO2021175271 A1 WO 2021175271A1
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substituted
cancer
unsubstituted
following group
group
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PCT/CN2021/078966
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French (fr)
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江磊
冯志勇
金贤
刘胜洋
石倩
寿建勇
徐圆
张建华
赵海霞
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轶诺(浙江)药业有限公司
上海轶诺药业有限公司
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Priority to CN202180019219.4A priority Critical patent/CN115244050A/zh
Publication of WO2021175271A1 publication Critical patent/WO2021175271A1/zh

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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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • A61P35/02—Antineoplastic agents specific for leukemia
    • 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
    • C07D217/00—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems
    • C07D217/22—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the nitrogen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

  • the present invention relates to the field of small molecule drugs. Specifically, the present invention relates to a kinase inhibitor and its preparation and use.
  • T cells, B cells and dendritic cells are powerful weapons used by the human immune system to resist foreign invaders such as viruses and bacteria, as well as self-infected cells or abnormal cells such as cancerous cells.
  • the human body has a complex and sophisticated control system to ensure the normal operation of the immune system.
  • cancerous cells appear in the human body, if the immune system cannot completely kill them, these immune escaped cancer cells will proliferate abnormally and form tumors.
  • Traditional tumor treatment is mainly through surgery, radiotherapy, chemotherapy and molecular targeted drugs. However, for many types of tumors or cancers, surgical resection is often not a viable option. While radiotherapy and chemotherapy target tumor cells, they also damage some healthy cells.
  • HPK1 Hematopoietic progenitor cell kinase 1
  • MAP4K1 a member of the MAP4K family
  • DCs dendritic cells
  • HPK1 is mainly expressed by hematopoietic cells, including early hematopoietic progenitor cells. In T cells, HPK1 is believed to be able to degrade these proteins by phosphorylating the Ser376 site of the downstream SLP76 protein and the Thr254 site of the Gads protein and recruiting 14-3-3 protein, thereby reducing the persistence of the signaling microclusters. To negatively regulate the role of T cell activation. HPK1 can also be activated in response to prostaglandins (PGE2) normally secreted by tumors, helping tumor cells escape from the immune system.
  • PGE2 prostaglandins
  • HPK1 can also inhibit AP-1, and AP-1 plays a role in promoting cell proliferation, inhibiting differentiation, and promoting tumor cell invasion and metastasis during tumor formation and development.
  • Targeted destruction of HPK1 kinase alleles can enable T cells to increase the production of Th1 cytokines (IL-2, IFN ⁇ , etc.) in the TCR response.
  • HPK1 has multiple roles in immunity and is related to the pathogenesis of autoimmune diseases, cancer and inflammation.
  • HPK1 kinase knockout (HPK1-/-) T cells proliferate much faster than monomeric wild-type, and mice transfected with HPK1-/-T cells can resist tumor growth, and dendritic cells (DCs) that have lost HPK1 kinase ) Compared with wild type, it has better antigen presentation ability and can better show anti-tumor immune response.
  • DCs dendritic cells
  • HPK1 kinase knockout T cells proliferate much faster than monomeric wild-type, and mice transfected with HPK1-/-T cells can resist tumor growth, and dendritic cells (DCs) that have lost HPK1 kinase )
  • DCs dendritic cells
  • HPK1 kinase knockout T cells proliferate much faster than monomeric wild-type, and mice transfected with HPK1-/-T cells can resist tumor growth, and dendritic cells (DCs) that have lost HPK1 kinase )
  • the purpose of the present invention is to provide a small molecule HPK1 inhibitor with selectivity and high activity.
  • the first aspect of the present invention provides a compound represented by the following formula I, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof;
  • X is selected from N or CH, wherein the CH may be further substituted by R 1;
  • Ar ring is selected from the following group: 6-10 membered aromatic ring, or 5-10 membered heteroaromatic ring;
  • Ring B is selected from the following group: 6-12 membered aromatic ring, or 5-12 membered heteroaromatic ring; or 8-15 membered bicyclic ring containing ring structure unit selected from the following group: 6-10 membered aromatic ring , 5-10 membered heteroaromatic ring, 3-10 membered heterocyclic group, C3-C8 cycloalkyl;
  • n 0, 1, 2 or 3;
  • n and p are each independently 0, 1, 2, 3, or 4;
  • Re is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted one with 1-3 heteroatoms selected from N, S and O 3-10 membered heterocyclic group; wherein, the substitution preferably includes one or more groups selected from the following group: -N(Rc) 2
  • Rc is selected from the following group: H, or substituted or unsubstituted C1-C5 alkyl
  • R 7 , R 8 , R 9 , R 10 , R 11 , and R 12 is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy , Substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group with 1-3 heteroatoms selected from the following group of N, S and O;
  • each chiral center is R configuration or S configuration.
  • the Re is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 3- 3 heteroatoms selected from N, S and O. 10-membered heterocyclic group.
  • the Ar ring is a 5- to 7-membered heteroaromatic ring; and the R 2 is selected from the group consisting of H, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted Substituted C1-C6 alkoxy.
  • the compound of formula I has the structure shown in formula II as follows:
  • Ra and Rb are each independently selected from the following group: hydrogen, halogen, CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;
  • Rb and R 4 together form a ring atom attached to a 5-6 membered carbocyclic or heterocyclic ring, preferably, the carbocyclic or heterocyclic non-aromatic ring.
  • the compound of formula I has the structure shown in the following formula III:
  • the compound of formula I is selected from the following group:
  • a pharmaceutical composition comprising (1) the compound according to claim 1 or its stereoisomer or tautomer, or its pharmaceutically Acceptable salts, hydrates or solvates; (2) pharmaceutically acceptable carriers.
  • the third aspect of the present invention provides a compound as described in the first aspect of the present invention or a stereoisomer or tautomer, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or
  • the use of the pharmaceutical composition according to the second aspect of the invention is to prepare a pharmaceutical composition for preventing and/or treating diseases related to the activity or expression of HPK1 kinase.
  • the fourth aspect of the present invention provides a compound according to the first aspect of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates and deuterated compounds and other tumor immunotherapy
  • the other tumor immunotherapeutic agents are selected from the following group: small molecule compounds and antibodies (including but not limited to PD-1, PD-L1, CTLA-4, TIM-3, TGF- ⁇ and Its receptors, LAG3 antagonists or TLR4, TLR7, TLR8, TLR9, STING agonists, etc.), chemotherapy regimens, radiotherapy regimens, tumor-targeted drugs, tumor vaccines, etc.
  • the diseases include but are not limited to cancer, metastatic cancer, inflammation and autoimmune related diseases.
  • the disease includes but is not limited to: lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuroendocrine tumor, carcinoid tumor, Gastrinoma, islet cell carcinoma, mesothelioma, schwannoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignancies, squamous cell carcinoma, epithelial squamous cell carcinoma, lung cancer, small Cell lung cancer, non-small cell lung cancer, adenocarcinoma, lung cancer, lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastric cancer, bowel cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, breast cancer Cancer, metastatic
  • the term “about” means that the value can vary from the recited value by no more than 5%.
  • the expression “about 100” includes all values between 95 and 105 (eg, 95.1, 95.2, 95.3, 95.4, etc.).
  • the term "containing” or “including (including)” can be open, semi-closed, and closed. In other words, the term also includes “substantially consisting of” or “consisting of”.
  • alkyl includes straight or branched chain alkyl groups.
  • C 1 -C 8 alkyl means a straight or branched alkyl group having 1-8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl Wait.
  • alkenyl includes linear or branched alkenyl.
  • C 2 -C 6 alkenyl refers to a linear or branched alkenyl having 2-6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2 -Butenyl, or similar groups.
  • alkynyl includes straight-chain or branched alkynyl groups.
  • C 2 -C 6 alkynyl refers to a linear or branched alkynyl group having 2-6 carbon atoms, such as ethynyl, propynyl, butynyl, or the like.
  • C 3 -C 8 cycloalkyl refers to a cycloalkyl group having 3-8 carbon atoms. It may be a monocyclic ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or similar groups. It may also be in the form of a bicyclic ring, such as a bridged ring or a spiro ring.
  • C 1 -C 8 alkoxy refers to a linear or branched alkoxy group having 1-8 carbon atoms; for example, methoxy, ethoxy, propoxy, iso Propoxy, butoxy, isobutoxy, tert-butoxy, etc.
  • the term "3-10 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S, and O" refers to a group having 3-10 atoms and 1-3 atoms are selected Saturated or partially saturated cyclic groups of heteroatoms from the following group of N, S, and O. It can be a single ring or a double ring form, such as a bridged ring or a spiro ring.
  • C 6 -C 10 aryl group refers to an aryl group having 6-10 carbon atoms, for example, a phenyl group or a naphthyl group and the like.
  • N, S, and O refers to those having 5-10 atoms and wherein 1-3 atoms are selected from The following group of N, S, and O heteroatoms are cyclic aromatic groups. It may be a monocyclic ring or a condensed ring form.
  • pyridyl pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2, 4)-Triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, etc.
  • the groups of the present invention can be substituted by substituents selected from the group consisting of halogen, nitrile, nitro, hydroxyl, amino, C 1 -C 6 alkyl-amino, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 alkoxy, halo C 1 -C 6 alkyl, halo C 2 -C 6 alkenyl, halogen Substituted C 2 -C 6 alkynyl, halogenated C 1 -C 6 alkoxy, allyl, benzyl, C 6 -C 12 aryl, C 1 -C 6 alkoxy-C 1 -C 6 alkane Group, C 1 -C 6 alkoxy-carbonyl, phenoxycarbonyl, C 2 -C 6 alkynyl-carbonyl, C 2 -C 6 alkenyl-carbonyl, C
  • halogen or halogen atom refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl and Br. "Halogenated” refers to substitution with an atom selected from F, Cl, Br, and I.
  • the structural formula described in the present invention is intended to include all isomeric forms (such as enantiomers, diastereomers and geometric isomers (or conformational isomers)): for example, containing asymmetry The R and S configuration of the center, the (Z) and (E) isomers of the double bond, etc. Therefore, a single stereochemical isomer of the compound of the present invention or a mixture of its enantiomers, diastereomers or geometric isomers (or conformational isomers) all belong to the scope of the present invention.
  • tautomers means that structural isomers with different energies can exceed the low energy barrier to convert into each other.
  • proton tautomers ie, proton transfer
  • Valence tautomers include interconversion through the recombination of some bonding electrons.
  • solvate refers to a complex in which the compound of the present invention coordinates with solvent molecules to form a specific ratio.
  • the present invention provides a compound represented by the following formula I:
  • X is selected from N or CH, wherein the CH may be further substituted by R 1;
  • Ar ring is selected from the following group: 6-10 membered aromatic ring, or 5-10 membered heteroaromatic ring;
  • Ring B is selected from the following group: 6-12 membered aromatic ring, or 5-12 membered heteroaromatic ring; or 8-15 membered bicyclic ring containing ring structure unit selected from the following group: 6-10 membered aromatic ring , 5-10 membered heteroaromatic ring, 3-10 membered heterocyclic group, C3-C8 cycloalkyl;
  • n 0, 1, 2 or 3;
  • n and p are each independently 0, 1, 2, 3, or 4;
  • Re is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted one with 1-3 heteroatoms selected from N, S and O 3-10 membered heterocyclic group; wherein, the substitution preferably includes one or more groups selected from the following group: -N(Rc) 2 ;
  • Rc is selected from the following group: H, or substituted or unsubstituted C1-C5 alkyl
  • R 7 , R 8 , R 9 , R 10 , R 11 , and R 12 is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy , Substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12 membered heterocyclic group with 1-3 heteroatoms selected from the following group of N, S and O;
  • each chiral center is R configuration or S configuration.
  • Carboxy group or substituted or unsubstituted group selected from the following group: C1-C6 alkyl group, C6-C10 aryl group, 5-10 member with 1-3 heteroatoms selected from N, S and O Heteroaryl, and the substituent is selected from the following group: halogen, C1-C6 alkyl, C1-C6 alkylene-OH, C1-C6 alkoxy, oxo, -S(O) 2 CH 3 , -CN, -OH, C6-C10 aryl, 3-10 membered heteroaryl with 1-3 heteroatoms selected from N, S and O, -C(O)CHNH 2 , -C(O) CHOH.
  • the X, Ar, B, m, n, p, Re, Rc, Rf, R 1 , R 2 , R 3 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 Each is independently the corresponding group in the specific compound in each example.
  • the compounds of the present invention can be used as HPK1 kinase inhibitors, in a preferred embodiment, HPK1 kinase inhibitors.
  • the compound of formula I of the present invention can be prepared by the following method:
  • a brominated reagent is used to react with boric acid to obtain a compound of formula I;
  • a brominated reagent is used to react with the borate to obtain the compound of formula I.
  • the compound of the present invention has excellent HPK1 kinase inhibitory activity, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and containing the compound of the present invention as the main active ingredient
  • the pharmaceutical composition can be used to prevent and/or treat diseases related to the activity or expression of HPK1 kinase (e.g., cancer).
  • the pharmaceutical composition of the present invention contains a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable excipient or carrier.
  • the "safe and effective amount” refers to: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects.
  • the pharmaceutical composition contains 1-2000 mg of the compound of the present invention/agent, more preferably, 10-200 mg of the compound of the present invention/agent.
  • the "one dose” is a capsule or tablet.
  • “Pharmaceutically acceptable carrier” refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use, and must have sufficient purity and sufficiently low toxicity. "Compatibility” here means that the components in the composition can be blended with the compound of the present invention and between them without significantly reducing the efficacy of the compound.
  • pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, and solid lubricants (such as stearic acid).
  • Magnesium stearate calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.
  • vegetable oils such as soybean oil, sesame oil, peanut oil, olive oil, etc.
  • polyols such as propylene glycol, glycerin, mannitol, sorbitol, etc.
  • emulsifiers such as emulsifiers
  • wetting agents such as sodium lauryl sulfate
  • the method of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous) injection, and the like.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.
  • the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with the following ingredients: (a) fillers or compatibilizers, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) Absorption accelerators, such as quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glycty
  • Solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the active compound or the release of the compound in such a composition may be released in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures.
  • the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-Butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.
  • composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
  • suspending agents for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
  • composition for parenteral injection may contain physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.
  • the compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.
  • the pharmaceutical composition When administered in combination, the pharmaceutical composition also includes one or more other pharmaceutically acceptable compounds.
  • One or more of the other pharmaceutically acceptable compounds may be administered simultaneously, separately or sequentially with the compound of the present invention.
  • a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage when administered is the effective dosage considered pharmaceutically, for a person weighing 60 kg,
  • the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg.
  • the specific dosage should also consider factors such as the route of administration and the patient's health status, which are all within the skill range of a skilled physician.
  • the enzyme activity test buffer contains 40mM Tris, pH 7.5; 20mM mg Cl2; 0.1 mg/ml BSA; 50 ⁇ M DTT.
  • the compound is dissolved in 100% DMSO, and the concentration of the mother liquor is 10 mM.
  • the DMSO solution of the compound starts from 100uM, and it is continuously diluted three-fold, with a total of eleven concentrations.
  • the enzyme activity test buffer contains 40mM Tris, pH 7.5; 20mM mg Cl2; 0.1 mg/ml BSA; 50 ⁇ M DTT.
  • the compound is dissolved in 100% DMSO, and the concentration of the mother liquor is 10 mM.
  • the DMSO solution of the compound is continuously diluted in three-fold gradient from 1 mM, with a total of eleven concentrations. After the diluted compound is diluted 1:20 with enzyme activity test buffer, 1uL is added to the working well, each concentration is two Multiple holes. Add 1uL of DMSO solution diluted 1:20 to the negative control wells and the positive control wells.
  • the preparation of 2.5 ⁇ substrate/ATP working solution is an enzyme activity test buffer containing 0.5ug/uL PKA substrate peptide and 105uM ATP, and 2uL 2.5 ⁇ substrate/ATP working solution is added to each working well.
  • A represents the IC 50 values ⁇ 50nM
  • B represents 50nM ⁇ IC 50 values ⁇ 500nM
  • C represents 500nM ⁇ IC 50 values ⁇ 10uM.
  • Inhibition of HPK1 can inhibit the phosphorylation of downstream SLP76.
  • the phosphorylation of SLP76 protein uses Jurkat (ATCC, Clone E6-1 TIB-152 TM) cells were tested, the first day of the experiment cells were diluted with culture medium (RPMI 1640 + 0.5% FBS) to 106 / ml per well 100uL, 10 5 cells were plated in an amount of 96-well cell culture In the plate, starve for 4 hours and incubate. The compound is dissolved in 100% DMSO, and the concentration of the mother liquor is 4mM. The DMSO solution of the compound is continuously diluted in four times from 10 mM to a total of 9 concentrations.
  • 4 uL of the diluted compound is diluted to 196 uL of 37°C preheated RPMI 1640 and mixed.
  • Add 50uL of the final diluted compound to the cells incubate at 37°C for 20 minutes, and add 50uL of diluted human CD3/CD28T cell activator (Stemcell, catalog number: 10971) to make the final concentration volume of the activator equal to 1/ of the total system.

Abstract

本发明提供了一种HPK1激酶抑制剂及其制备和用途。具体地，本发明提供了一种如式I所示的化合物，其中，各基团的定义如说明书中所述。所述的化合物具有优异的HPK1抑制活性，因此可用于制备治疗癌症和其他HPK1活性相关疾病的药物组合物。

Description

新型HPK1抑制剂及其制备方法和应用 技术领域
本发明涉及小分子药物领域，具体地，本发明涉及一种激酶抑制剂及其制备和用途。
背景技术
T细胞、B细胞和树突状细胞(DCs)是人体免疫系统用于抵抗外来入侵者如病毒和细菌以及自身感染细胞或异常细胞如癌变细胞等的强有力武器。而人体存在一套复杂且精密的调控系统用于确保免疫系统的正常运行。当人体中出现癌变细胞的时候，如果免疫系统无法将其完全杀死，这些免疫逃逸的癌细胞异常增殖就会形成肿瘤。传统肿瘤治疗主要通过手术、放疗、化疗和分子靶向药物。然而，对于许多形式多样的肿瘤或癌症，手术切除往往并不是可行的选择。而放射疗法和化疗在靶向肿瘤细胞的同时，也会损害部分健康细胞。肿瘤细胞易于发生突变，并且可能对特异靶向肿瘤细胞的药物产生耐药性，使得癌症治疗困难重重。近年来利用患者自身的免疫系统来克服肿瘤细胞所采用的免疫逃逸策略，并增强机体抗肿瘤免疫力是一种新型的癌症治疗策略。其中一种策略是通过抑制通常起维持外周耐受作用的免疫反应的负调控因子，使肿瘤抗原被识别为非自身抗原，从而克服肿瘤细胞免疫逃逸。造血祖细胞激酶1(HPK1)又称MAP4K1(MAP4K家族成员)，是一个树突状细胞(DCs)、T细胞和B细胞活化反应的负调控因子，抑制其活性可以针对性地增强机体抗肿瘤免疫力。HPK1主要由造血细胞，包括早期造血祖细胞表达。在T细胞中，HPK1被认为能够通过磷酸化下游SLP76蛋白的Ser376位点和Gads蛋白的Thr254位点并招募14-3-3蛋白来降解这些蛋白，从而降低信号微团簇的持久性，起到负调控T细胞活化的作用。HPK1还可以通过响应通常由肿瘤分泌的前列腺素(PGE2)而被激活，有助于肿瘤细胞从免疫系统逃逸。并且HPK1还能抑制AP-1，而AP-1在肿瘤形成及发展过程中，在促进细胞增殖、抑制分化、促进肿瘤细胞的侵袭和转移等过程中发挥作用。针对性地破坏HPK1激酶的等位基因可以使T细胞在TCR应答中提高Th1细胞因子(IL-2，IFNγ等)的产生。HPK1在免疫中具有多种作用，并与自身免疫性疾病，癌症和炎症反应的发病机理有关。HPK1激酶敲除(HPK1-/-)的T细胞增殖相对于单体野生型快很多，并且转染过HPK1-/-T细胞的老鼠能抵抗肿瘤的生长，失去HPK1激酶的树突细胞(DCs)与野生型相比具有更好的抗原提呈能力，能更好地表现出抗肿瘤免疫应答。此外，动物实验研究表明，HPK1的抑制和PD-1/PD-L1抗体药物具有明显的协同抗肿瘤活性。因此，HPK1激酶在疾病治疗特别是癌症治疗中具有关键作用。
目前针对该靶点尚未有药物上市，为了满足未来临床的巨大需求，我们希望通过设计开发出具有选择性和高活性的小分子HPK1抑制剂，为免疫相关疾病特别是肿瘤治疗提供新型的口服类药物，单独用药或者联合化疗、放疗、肿瘤靶向药、其他肿瘤免疫治疗剂(小分子化合物及抗体)以及肿瘤疫苗等。
发明内容
本发明的目的是提供一种具有选择性和高活性的小分子HPK1抑制剂。
本发明的第一方面，提供了一种如下式I所示的化合物，或其药学上可接受的盐、光学异构体或水合物；
Figure PCTCN2021078966-appb-000001
其中，
X选自N或CH，其中，所述的CH可以进一步被R 1取代；
Ar环选自下组：6-10元芳香环，或5-10元的杂芳环；
B环选自下组：6-12元芳香环，或5-12元的杂芳环；或含有选自下组的环结构单元的8-15元二环并环：6-10元芳香环、5-10元的杂芳环、3-10元杂环基、C3-C8环烷基；
m为0、1、2或3；
n和p各自独立地为0、1、2、3或4；
Re选自下组：取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自N、S和O的杂原子的3-10元杂环基；其中，所述的取代优选包括一个或多个选自下组的基团：-N(Rc) 2
Rc选自下组：H、或取代或未取代的C1-C5的烷基；
Rf选自下组：氢、卤素、CN、取代或未取代的C1-C6烷基、-NH 2、R 9R 10N-C(＝O)-；
各个R 1、R 2、R 3和Rg各自独立地选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-12元杂环基、取代或未取代的C6-C10芳基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-10元杂芳基、R 7-C(＝O)-、R 8-S(＝O) 2-、R 9R 10N-C(＝O)-、R 9R 10C(＝O)N(Rc)-、R 11R 12N-S(＝O) 2-；
各个R 7、R 8、R 9、R 10、R 11、R 12各自独立地选自下组：H、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-12元杂环基；
除非特别说明，所述的“取代”是指被选自下组的一个或多个(例如2个、3个、4个等)取代基所取代：卤素、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、甲基砜基、氧代(＝O)、-CN、羟基、羧基、C1-C6酰胺基(-C(＝O)-N(Rc) 2或-NH-C(＝O)(Rc))、-N(Rc) 2、或取代或未取代的选自下组的基团：C1-C6烷基、C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、-(CH 2)-C6-C10芳基、-(CH 2)-(具有1-3个选自N、S和O的杂原子的5-10元杂芳基)、-(具有1-3个选自N、S和O的杂原子的5-10元亚杂芳基)-(C1-C6烷基)、具有1-3个选自下组N、S和O的杂原子的3-12元杂环基(包括单环、螺环、桥环或并环)，且所述的取代基选自下组：卤素、C1-C6烷基、C1-C6亚烷基-OH、C1-C6烷氧基、氧代、-S(O) 2CH 3、-CN、-OH、C6-C10芳基、具有1-3个选自N、S和O的杂原子的3-10元杂芳基、-C(O)CHNH 2、-C(O)CHOH；
且所述的式I化合物中，各个手性中心为R构型或S构型。
在另一优选例中，所述的“取代”是指被选自下组的一个或多个(例如2个、3个、4个等)取代基所取代：卤素、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的 C3-C8环烷基、甲基砜基、氧代(＝O)、-CN、羟基、-NH 2、C1-C6胺基、羧基、或取代或未取代的选自下组的基团：C1-C6烷基、C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、且所述的取代基选自下组：卤素、C1-C6烷基、C1-C6亚烷基-OH、C1-C6烷氧基、氧代、-S(O) 2CH 3、-CN、-OH、C6-C10芳基、具有1-3个选自N、S和O的杂原子的3-10元杂芳基、-C(O)CHNH 2、-C(O)CHOH。
在另一优选例中，所述的Re选自下组：取代或未取代的C1-C6烷基、取代或未取代的具有1-3个选自N、S和O的杂原子的3-10元杂环基。
在另一优选例中，Ar环为5-7元的杂芳环；且所述的R 2选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基。
在另一优选例中，所述的B环选自下组：苯环、含有苯环结构单元的8-15元二环并环、5-6元的杂芳环、含有5-6元杂芳环结构单元的8-15元二环并环；其中，所述的R 3选自下组：H、卤素、取代或未取代的C1-C6烷基、取代的-C(＝O)-；其中，所述的取代优选包括一个或多个选自下组的基团：-N(Rc) 2。
在另一优选例中，所述的式I化合物具有如下式II所示的结构：
Figure PCTCN2021078966-appb-000002
其中，
所述的Ra和Rb各自独立地选自下组：氢、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；
所述的R 4选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、R 9R 10N-C(＝O)-；其中，所述的R 9、R 10选自下组：H、取代或未取代的C1-C6烷基；
或所述的Rb和R 4与相连的环原子共同构成5-6元碳环或杂环，较佳地，所述的碳环或杂环为非芳香性环。
在另一优选例中，所述的式I化合物具有如下式III所示的结构：
Figure PCTCN2021078966-appb-000003
其中，
所述的Rd选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的 C1-C6烷氧基、R 9R 10N-C(＝O)-；其中，所述的R 9、R 10选自下组：H、取代或未取代的C1-C6烷基。
在另一优选例中，所述的式I化合物选自下组：
Figure PCTCN2021078966-appb-000004
Figure PCTCN2021078966-appb-000005
本发明的第二方面，提供了一种药物组合物，所述的药物组合物包括(1)如权利要求1所述的化合物或其立体异构体或互变异构体，或其药学上可接受的盐、水合物或溶剂化物；(2)药学上可接受的载体。
本发明的第三方面，提供了如本发明第一方面所述的化合物或其立体异构体或互变异构体，或其药学上可接受的盐、水合物或溶剂化物，或如本发明第二方面所述的药物组合物的用途，用于制备预防和/或治疗与HPK1激酶的活性或表达量相关的疾病的药物组合物。
本发明的第四方面，提供了一种本发明第一方面所述的化合物及其药学上可接受的盐、立体异构体、酯、前药、溶剂化物和氘代化合物与其他肿瘤免疫治疗剂的联合肿瘤化疗方案，所述的其他肿瘤免疫治疗剂选自下组：小分子化合物及抗体(包括但不限于PD-1、PD-L1、CTLA-4、TIM-3、TGF-β及其受体、LAG3拮抗剂或TLR4、TLR7、TLR8、TLR9、STING激动剂等)、化疗方案、放疗方案、肿瘤靶向药、肿瘤疫苗等。
在另一优选例中，所述的疾病包括但不限于癌症、转移性癌症、炎症和自身免疫相关疾病。
在另一优选例中，所述疾病包括但不限于：淋巴瘤，母细胞瘤，髓母细胞瘤，视网膜母细胞瘤，肉瘤，脂肪肉瘤，滑膜细胞肉瘤，神经内分泌肿瘤，类癌肿瘤，胃泌素瘤，胰岛细胞癌，间皮瘤，神经鞘瘤，听神经瘤，脑膜瘤，腺癌，黑色素瘤，白血病或淋巴样 恶性肿瘤，鳞状细胞癌，上皮鳞状细胞癌，肺癌，小细胞肺癌，非小细胞肺癌，腺癌肺癌，肺鳞癌，腹膜癌，肝细胞癌，胃癌，肠癌，胰腺癌，成胶质细胞瘤，子宫颈癌，卵巢癌，肝癌，膀胱癌，乳腺癌，转移性乳腺癌，结肠癌，直肠癌，结直肠癌，子宫癌，唾液腺癌，肾癌，前列腺癌，外阴癌，甲状腺癌，肛门癌，阴茎癌，梅克尔细胞癌，食管癌，胆道肿瘤，头颈部癌，血液恶性肿瘤，鼻咽癌，多发性骨髓瘤，大场绒毛腺瘤，非霍奇金淋巴瘤，骨癌，睾丸癌，霍奇金病，精元细胞瘤，口腔癌，脑癌，皮肤癌，乳腺导管癌，肾盂癌，肾母细胞瘤，食管腺瘤，视网膜细胞瘤，神经胶质瘤，神经纤维瘤，胃肠道间质瘤，原位癌，子宫内膜癌和骨髓增生异常综合征等。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
本发明人经过长期而深入的研究，设计并合成了一类新型HPK1激酶抑制剂。在此基础上，发明人完成了本发明。
术语
除非另外定义，否则本文中所用的全部技术与科学术语均具有如本发明所属领域的普通技术人员通常理解的相同含义。
如本文所用，在提到具体列举的数值中使用时，术语“约”意指该值可以从列举的值变动不多于5％。例如，如本文所用，表述“约100”包括95和105和之间的全部值(例如，95.1、95.2、95.3、95.4等)。
如本文所用，术语“含有”或“包括(包含)”可以是开放式、半封闭式和封闭式的。换言之，所述术语也包括“基本上由…构成”、或“由…构成”。
定义
如本文所用，术语“烷基”包括直链或支链的烷基。例如C 1-C 8烷基表示具有1-8个碳原子的直链或支链的烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基等。
如本文所用，术语“烯基”包括直链或支链的烯基。例如C 2-C 6烯基指具有2-6个碳原子的直链或支链的烯基，例如乙烯基、烯丙基、1-丙烯基、异丙烯基、1-丁烯基、2-丁烯基、或类似基团。
如本文所用，术语“炔基”包括直链或支链的炔基。例如C 2-C 6炔基是指具有2-6个碳原子的直链或支链的炔基，例如乙炔基、丙炔基、丁炔基、或类似基团。
如本文所用，术语“C 3-C 8环烷基”指具有3-8个碳原子的环烷基。其可以是单环，例如环丙基、环丁基、环戊基、环己基、或类似基团。也可以是双环形式，例如桥环或螺环形式。
如本文所用，术语“C 1-C 8烷氧基”是指具有1-8个碳原子的直链或支链的烷氧基；例如，甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、叔丁氧基等。
如本文所用，术语“具有1-3个选自下组N、S和O的杂原子的3-10元杂环基”是指具有3-10个原子的且其中1-3个原子为选自下组N、S和O的杂原子的饱和或部分饱和的环状基团。其可以是单环，也可以是双环形式，例如桥环或螺环形式。具体的 实例可以为氧杂环丁烷、氮杂环丁烷、四氢-2H-吡喃基、哌啶基、四氢呋喃基、吗啉基和吡咯烷基、1,2,3,4-四氢异喹啉基等。
如本文所用，术语“C 6-C 10芳基”是指具有6-10个碳原子的芳基，例如，苯基或萘基等类似基团。
如本文所用，术语“具有1-3个选自下组N、S和O的杂原子的5-10元杂芳基”指具有5-10个原子的且其中1-3个原子为选自下组N、S和O的杂原子的环状芳香基团。其可以是单环，也可以是稠环形式。具体的实例可以为吡啶基、哒嗪基、嘧啶基、吡嗪基、三嗪基、吡咯基、吡唑基、咪唑基、(1,2,3)-三唑基以及(1,2,4)-三唑基、四唑基、呋喃基、噻吩基、异噁唑基、噻唑基、噁唑基等。
除非特别说明，否则本发明的基团均可被选自下组的取代基所取代：卤素、腈基、硝基、羟基、氨基、C 1-C 6烷基-胺基、C 1-C 6烷基、C 2-C 6烯基、C 2-C 6炔基、C 1-C 6烷氧基、卤代C 1-C 6烷基、卤代C 2-C 6烯基、卤代C 2-C 6炔基、卤代C 1-C 6烷氧基、烯丙基、苄基、C 6-C 12芳基、C 1-C 6烷氧基-C 1-C 6烷基、C 1-C 6烷氧基-羰基、苯氧羰基、C 2-C 6炔基-羰基、C 2-C 6烯基-羰基、C 3-C 6环烷基-羰基、C 1-C 6烷基-磺酰基等。
如本文所用，“卤素”或“卤原子”指F、Cl、Br、和I。更佳地，卤素或卤原子选自F、Cl和Br。“卤代的”是指被选自F、Cl、Br、和I的原子所取代。
除非特别说明，本发明所描述的结构式意在包括所有的同分异构形式(如对映异构，非对映异构和几何异构体(或构象异构体))：例如含有不对称中心的R、S构型，双键的(Z)、(E)异构体等。因此，本发明化合物的单个立体化学异构体或其对映异构体、非对映异构体或几何异构体(或构象异构体)的混合物都属于本发明的范围。
如本文所用，术语“互变异构体”表示具有不同能量的结构同分异构体可以超过低能垒，从而互相转化。比如，质子互变异构体(即质子移变)包括通过质子迁移进行互变，如1H-吲唑与2H-吲唑。化合价互变异构体包括通过一些成键电子重组而进行互变。
如本文所用，术语“溶剂合物”是指本发明化合物与溶剂分子配位形成特定比例的配合物。
式I化合物
本发明提供了一种如下式I所示的化合物：
Figure PCTCN2021078966-appb-000006
其中，
X选自N或CH，其中，所述的CH可以进一步被R 1取代；
Ar环选自下组：6-10元芳香环，或5-10元的杂芳环；
B环选自下组：6-12元芳香环，或5-12元的杂芳环；或含有选自下组的环结构单元的8-15元二环并环：6-10元芳香环、5-10元的杂芳环、3-10元杂环基、C3-C8环烷 基；
m为0、1、2或3；
n和p各自独立地为0、1、2、3或4；
Re选自下组：取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自N、S和O的杂原子的3-10元杂环基；其中，所述的取代优选包括一个或多个选自下组的基团：-N(Rc) 2；
Rc选自下组：H、或取代或未取代的C1-C5的烷基；
Rf选自下组：氢、卤素、CN、取代或未取代的C1-C6烷基、-NH 2、R 9R 10N-C(＝O)-；
各个R 1、R 2R 3和Rg各自独立地选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-12元杂环基、取代或未取代的C6-C10芳基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-10元杂芳基、R 7-C(＝O)-、R 8-S(＝O) 2-、R 9R 10N-C(＝O)-、R 9R 10C(＝O)N(Rc)-、R 11R 12N-S(＝O) 2-；
各个R 7、R 8、R 9、R 10、R 11、R 12各自独立地选自下组：H、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-12元杂环基；
除非特别说明，所述的“取代”是指被选自下组的一个或多个(例如2个、3个、4个等)取代基所取代：卤素、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、甲基砜基、氧代(＝O)、-CN、羟基、羧基、C1-C6酰胺基(-C(＝O)-N(Rc) 2或-NH-C(＝O)(Rc))、-N(Rc) 2或取代或未取代的选自下组的基团：C1-C6烷基、C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、-(CH 2)-C6-C10芳基、-(CH 2)-(具有1-3个选自N、S和O的杂原子的3-10元杂芳基)、-(具有1-3个选自N、S和O的杂原子的5-10元亚杂芳基)-(C1-C6烷基)、具有1-3个选自下组N、S和O的杂原子的3-12元杂环基(包括单环、螺环、桥环或并环)，且所述的取代基选自下组：卤素、C1-C6烷基、C1-C6亚烷基-OH、C1-C6烷氧基、氧代、-S(O) 2CH 3、-CN、-OH、C6-C10芳基、具有1-3个选自N、S和O的杂原子的3-10元杂芳基、-C(O)CHNH 2、-C(O)CHOH；
且所述的式I化合物中，各个手性中心为R构型或S构型。
在另一优选例中，所述的“取代”是指被选自下组的一个或多个(例如2个、3个、4个等)取代基所取代：卤素、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、甲基砜基、氧代(＝O)、-CN、羟基、-N(Rc) 2、羧基、或取代或未取代的选自下组的基团：C1-C6烷基、C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、且所述的取代基选自下组：卤素、C1-C6烷基、C1-C6亚烷基-OH、C1-C6烷氧基、氧代、-S(O) 2CH 3、-CN、-OH、C6-C10芳基、具有1-3个选自N、S和O的杂原子的3-10元杂芳基、-C(O)CHNH 2、-C(O)CHOH。
优选地，所述的X、Ar、B、m、n、p、Re、Rc、Rf、R 1、R 2、R 3、R 7、R 8、R 9、R 10、R 11、R 12各自独立地为各个实施例中具体化合物中的对应基团。
本发明的化合物可以作为HPK1激酶抑制剂，在优选的实施例中，为HPK1激酶抑制剂。
式I化合物的制备
本发明的式I化合物可以通过以下方法制备：
Figure PCTCN2021078966-appb-000007
在惰性溶剂中，用溴代试剂与硼酸反应，得到式I化合物；
或
Figure PCTCN2021078966-appb-000008
在惰性溶剂中，用溴代试剂与硼酸酯反应，得到式I化合物。
药物组合物和施用方法
由于本发明化合物具有优异的HPK1激酶的抑制活性，因此本发明化合物及其各种晶型，药学上可接受的无机或有机盐，水合物或溶剂合物，以及含有本发明化合物为主要活性成分的药物组合物可用于预防和/或治疗与HPK1激酶活性或表达量相关的疾病(例如，癌症)。
本发明的药物组合物包含安全有效量范围内的本发明化合物及药学上可以接受的赋形剂或载体。其中“安全有效量”指的是：化合物的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有1-2000毫克本发明化合物/剂，更佳地，含有10-200毫克本发明化合物/剂。较佳地，所述的“一剂”为一个胶囊或药片。
“药学上可接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份能和本发明化合物以及它们之间相互掺和，而不明显降低化合物的药效。药学上可以接受的载体部分例子有纤维素及其衍生物(如羧甲基纤维素钠、乙基纤维素钠、纤维素乙酸酯等)、明胶、滑石、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂(如
Figure PCTCN2021078966-appb-000009
)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。
本发明化合物或药物组合物的施用方式没有特别限制，代表性的施用方式包括(但并不限于)：口服、肠胃外(静脉内、肌肉内或皮下)注射等。
用于口服给药的固体剂型包括胶囊剂、片剂、丸剂、散剂和颗粒剂。在这些固体剂型中，活性化合物与至少一种常规惰性赋形剂(或载体)混合，如柠檬酸钠或磷酸二钙，或与下述成分混合：(a)填料或增容剂，例如，淀粉、乳糖、蔗糖、葡萄糖、甘露醇和硅酸；(b)粘合剂，例如，羟甲基纤维素、藻酸盐、明胶、聚乙烯基吡咯烷酮、蔗糖和阿拉伯胶；(c)保湿剂，例如，甘油；(d)崩解剂，例如，琼脂、碳酸钙、马铃薯淀粉或木薯淀粉、藻酸、某些复合硅酸盐、和碳酸钠；(e)缓溶剂，例如石蜡；(f)吸收加速剂，例如，季胺化合物；(g)润湿剂，例如鲸蜡醇和单硬脂酸甘油酯；(h)吸附 剂，例如，高岭土；和(i)润滑剂，例如，滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、十二烷基硫酸钠，或其混合物。胶囊剂、片剂和丸剂中，剂型也可包含缓冲剂。
固体剂型如片剂、糖丸、胶囊剂、丸剂和颗粒剂可采用包衣和壳材制备，如肠衣和其它本领域公知的材料。它们可包含不透明剂，并且，这种组合物中活性化合物或化合物的释放可以延迟的方式在消化道内的某一部分中释放。可采用的包埋组分的实例是聚合物质和蜡类物质。必要时，活性化合物也可与上述赋形剂中的一种或多种形成微胶囊形式。
用于口服给药的液体剂型包括药学上可接受的乳液、溶液、悬浮液、糖浆或酊剂。除了活性化合物外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例知，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。
除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂、甜味剂、矫味剂和香料。
除了活性化合物外，悬浮液可包含悬浮剂，例如，乙氧基化异十八烷醇、聚氧乙烯山梨醇和脱水山梨醇酯、微晶纤维素、甲醇铝和琼脂或这些物质的混合物等。
用于肠胃外注射的组合物可包含生理上可接受的无菌含水或无水溶液、分散液、悬浮液或乳液，和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的含水和非水载体、稀释剂、溶剂或赋形剂包括水、乙醇、多元醇及其适宜的混合物。
本发明化合物可以单独给药，或者与其他药学上可接受的化合物联合给药。
联合给药时，所述药物组合物还包括与一种或多种其他药学上可接受的化合物。该其他药学上可接受的化合物中的一种或多种可与本发明的化合物同时、分开或顺序地给药。
使用药物组合物时，是将安全有效量的本发明化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量为药学上认为的有效给药剂量，对于60公斤体重的人而言，日给药剂量通常为1～2000毫克，优选20～500毫克。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。
各实施例中：
分析方法I
LCMS仪器：waters Acquity UPLC-MS，UV检测器：Acquity UPLC
色谱柱：Acquity UPLC HSS T3 1.8uM,柱温40℃
流动相：A：H2O(0.1％TFA)，B：乙腈，梯度洗脱。
以下实施例中所用的实验材料和试剂如无特别说明均可从市售渠道获得。
实施例1：2-(2-氟-6-甲基苯基)-8-(6-(4-甲基哌嗪-1-基)吡啶-3-基)-1,6-二氮杂萘-5-胺
Figure PCTCN2021078966-appb-000010
5-氯-1,6-萘啶-1-氧化物
Figure PCTCN2021078966-appb-000011
向配有磁力搅拌器的50毫升茄形瓶中加入5-氯-1,6-二氮杂萘(500毫克，3.04毫摩尔)和氯仿(10毫升)，将该反应液置于冰浴中冷却20分钟。向溶液中分批加入间氯过氧苯甲酸(纯度：85％，926毫克，4.56毫摩尔)。然后将反应混合物置于室温搅拌过夜。反应经LCMS检测。反应结束后，反应液经减压浓缩后得固体残留物，该残留物通过快速硅胶柱(二氯甲烷:甲醇＝100:1)分离,得到黄色固体5-氯-1,6-萘啶-1-氧化物(597毫克，纯度：90％，收率：100％)。MS(ESI):m/z＝181.0,183.0[M+H] +.
5-氯-1,6-二氮杂萘-2(1H)-酮
Figure PCTCN2021078966-appb-000012
在冰浴冷却下，向5-氯-1,6-萘啶-1-氧化物(450毫克，2.5毫摩尔)和三乙胺(759毫克，7.5毫摩尔)的无水四氢呋喃溶液(30毫升)中缓缓滴加三氟乙酸酐(3.15克，15毫摩尔)。滴加完毕后，将该混合物于0度继续搅拌1.5小时，然后将反应混合物升至室温，继续搅拌过夜。反应经LCMS检测。反应结束后将反应液缓慢倒入饱和碳酸氢钠水溶液(90毫升)中，用然后用二氯甲烷萃取(30毫升×3)。合并的有机相用无水硫酸钠干燥，抽滤，滤液经减压浓缩得粗品。该粗品经快速硅胶柱(石油醚:乙酸乙酯＝2:1-1:1)分离洗脱,得到白色固体5-氯-1,6-二氮杂萘-2(1H)-酮(200毫克，纯度：90％，收率：44％)。MS(ESI):m/z＝181.0,183.0[M+H] +.
1H NMR(400MHz,CDCl 3)δ8.33(d,J＝5.6Hz,1H),8.21(d,J＝10.0Hz,1H),7.20(d,J＝5.6Hz,1H),6.82(d,J＝9.6Hz,1H).
5-((二苯亚甲基)氨基)-1,6-二氮杂萘-2(1H)-酮
Figure PCTCN2021078966-appb-000013
在氮气保护下，向5-氯-1,6-二氮杂萘-2(1H)-酮(89毫克，0.494毫摩尔)，二苯甲酮亚胺(107毫克，0.593毫摩尔)和碳酸铯(322毫克，0.99毫摩尔)，的二氧六环(1.0毫升)混合溶液中加入醋酸钯(22毫克，0.099毫摩尔)，BINAP(61毫克，0.099毫摩尔)。将该混合物加热至120摄氏度搅拌过夜。反应经LCMS液相检测。反应结束后，反应液冷却至室温，过滤，滤液经减压浓缩得残留物。该残留物经快速硅胶柱(SiO 2，二氯甲烷:甲醇＝100:1-30:1)分离纯化得黄色固体5-((二苯亚甲基)氨基)-1,6-二氮杂萘-2(1H)-酮(90毫克，纯度：90％，收率：55％)。MS(ESI):m/z＝326.0[M+H] +.
5-((二苯亚甲基)氨基)-1,6-二氮杂萘-2-基三氟甲磺酸
Figure PCTCN2021078966-appb-000014
在冰水浴下，向5-((二苯亚甲基)氨基)-1,6-二氮杂萘-2(1H)-酮(430毫克，1.32毫摩尔)的无水二氯甲烷(10毫升)溶液中滴加三氟甲磺酸酐(683毫克，2,64毫摩尔)的二氯甲烷(1.0毫升)溶液。滴加完毕后，将该反应混合物于0度下继续搅拌60分钟。反应经LCMS检测。待原料转化完全后，将反应液倒入20毫升水中，然后用二氯甲烷(10毫升×2)萃取。合并的有机相经无水硫酸钠干燥，抽滤，滤液浓缩得粗品。该粗品经快速硅胶柱(石油醚:乙酸乙酯＝2:1)分离纯化，得5-((二苯亚甲基)氨基)-1,6-二氮杂萘-2-基三氟甲磺酸(420毫克，收率：75.6％)。MS(ESI):m/z＝458.0[M+H] +.
2-(2-氟-6-甲基苯基)-1,6-二氮杂萘-5-胺
Figure PCTCN2021078966-appb-000015
在氩气保护下，向5-((二苯亚甲基)氨基)-1,6-二氮杂萘-2-基三氟甲磺酸(420毫克，0.918毫摩尔)，2-(2-氟-6-甲基苯基)-4,4,5,5-四甲基-1,3,2-二噁硼戊环(238毫克，1.01毫摩尔)，磷酸钾(389毫克，1.84毫摩尔)和1,4-二氧六环(6.0毫升)/水(1.5毫升)的混合物中加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(74毫克,0.091毫摩尔)。将该反应液加热至100℃搅拌8小时。反应经LCMS检测。待反应结束后，反应液冷却至室温，加乙酸乙酯(10毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩溶剂得粗品，该粗品经快速硅胶柱(石油醚:乙酸乙酯＝1:1)分离得2-(2-氟-6-甲基苯基)-1,6-二氮杂萘-5-胺(270毫克)。MS(ESI):m/z＝254.1[M+H] +.
1H NMR(400MHz,CDCl 3)δ8.28(d,J＝8.8Hz,1H),8.12(d,J＝6.0Hz,1H),7.52(dd,J＝8.4,2.0Hz,1H),7.36–7.28(m,2H),7.12(d,J＝7.6Hz,1H),7.03(t,J＝8.8Hz,1H),2.26(s,3H).
8-溴-2-(2-氟-6-甲基苯基)-1,6-二氮杂萘-5-胺
Figure PCTCN2021078966-appb-000016
在冰浴冷却下，向2-(2-氟-6-甲基苯基)-1,6-二氮杂萘-5-胺(170毫克，0.67毫摩尔)的二氯甲烷(4.0毫升)溶液中分批加入N-溴代琥珀酰亚胺(125毫克，0.70毫摩尔)。将反应液于0度搅拌0.5小时后，撤去冰浴，于室温继续搅拌1小时。反应经LCMS检测。待反应结束后，向反应液中加入10％的亚硫酸钠(10毫升)水溶液中，搅拌10分钟后，分液收集有机向。水相用二氯甲烷(5毫升×2)萃取。合并的有机相经水(10毫升)洗涤，无水硫酸钠干燥，过滤，滤液经减压浓缩溶剂得粗品，该粗品经快速硅胶柱(石油醚:乙酸乙酯＝1:1)分离得到固体8-溴-2-(2-氟-6-甲基苯基)-1,6-二氮杂萘-5-胺(128毫克)。MS(ESI):m/z＝332.0,334.0.[M+H] +.
2-(2-氟-6-甲基苯基)-8-(6-(4-甲基哌嗪-1-基)吡啶-3-基)-1,6-二氮杂萘-5-胺
Figure PCTCN2021078966-appb-000017
在氩气保护下，向8-溴-2-(2-氟-6-甲基苯基)-1,6-二氮杂萘-5-胺(60毫克，0.18毫摩尔)，(6-(4-甲基哌嗪-1-基)吡啶-3-基)硼酸(48毫克，0.22毫摩尔)，磷酸钾(77毫克，0.36毫摩尔)和1,4-二氧六环(2.0毫升)/水(0.5毫升)的混合物中加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(14.6毫克,0.018毫摩尔)。将该反应液加热至100℃搅拌3小时。反应经LCMS检测。待反应结束后，反应液冷却至室温，加乙酸乙酯(10毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩溶剂得粗品，该粗品经Prep-HPLC(NH 4HCO 3)分离得2-(2-氟-6-甲基苯基)-8-(6-(4-甲基哌嗪-1-基)吡啶-3-基)-1,6-二氮杂萘-5-胺(26.3毫克，收率：30.1％)。MS(ESI):m/z＝429.1[M+H] +.
1H NMR(400MHz,CD 3OD)δ8.64(d,J＝8.8Hz,1H),8.37(d,J＝2.0Hz,1H),8.01(s,1H),7.86(dd,J＝8.8,2.4Hz,1H),7.55(dd,J＝8.8,2.0Hz,1H),7.35–7.27(m,1H),7.10(d,J＝8.0Hz,1H),7.02(t,J＝8.8Hz,1H),6.87(d,J＝8.8Hz,1H),3.59–3.49(m,4H),2.62–2.52(m,4H),2.34(s,3H),2.16(s,3H).
实施例2:1-氨基-6-(2-氟-6-甲基苯基)-4-(6-(4-甲基哌嗪-1-基)吡啶-3-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000018
6-氯异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000019
在氩气保护下，向化合物7-溴-6-氯异喹啉(900毫克，3.96毫摩尔)和氰化锌(232毫克，2.05毫摩尔)的N,N-二甲基甲酰胺(15毫升)混合物中,加入四-三苯基磷钯(457毫克，0.39毫摩尔)，反应液加热至100度搅拌2小时。反应经LCMS检测。待反应结束后，反应液冷却至室温，将反应液倒入水(50毫升)中，然后用乙酸乙酯(20毫升×2)萃取，合并有机层，依次用水(20毫升×2)，饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，过滤，滤液经减压浓缩得粗品。该粗品经快速硅胶柱(石油醚:乙酸乙酯＝5:1)分离纯化得到类白色固体6-氯异喹啉-7-甲腈(460毫克,收率：66％)。MS(ESI):m/z＝189.0,191.0[M+H] +.
6-(2-氟-6-甲基苯基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000020
在氩气保护下，向6-氯异喹啉-7-甲腈(460毫克，2.43毫摩尔)，2-(2-氟-6-甲基苯基)-4,4,5,5-四甲基-1,3,2-二噁硼戊环(689毫克，2.92毫摩尔)，碳酸钠(772毫克，7.29毫摩尔)和1,4-二氧六环(8.0毫升)/水(2.0毫升)的混合物中加入四-三苯基磷钯(280毫克,0.24毫摩尔)。将该反应液加热至100℃搅拌16小时。反应经LCMS检测。待反应结束后，反应液冷却至室温，加乙酸乙酯(10毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩溶剂得粗品，该粗品经快速硅胶柱(石油醚:乙酸乙酯 ＝3:1)分离得6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(360毫克，收率：57％)。MS(ESI):m/z＝263.0[M+H] +.
7-氰基-6-(2-氟-6-甲基苯基)异喹啉-2-氧化物
Figure PCTCN2021078966-appb-000021
向配有磁力搅拌器的50毫升茄形瓶中加入6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(360毫克，1.37毫摩尔)的氯仿(10毫升)溶液，将反应液置于冰浴中冷却。然后向溶液中分批加入间氯过氧苯甲酸(纯度：85％，353毫克，2.01毫摩尔)。然后将反应混合物置于室温搅拌过夜。反应经LCMS检测。待反应结束后，将反应液倒入冰水(20毫升)中，然后用二氯甲烷(10毫升×2)萃取。合并的有机相依次经饱和碳酸氢钠溶液(15毫升)，水(15毫升)，饱和食盐水(15毫升)洗涤，无水硫酸钠干燥。过滤，滤液经减压浓缩后得粗品7-氰基-6-(2-氟-6-甲基苯基)异喹啉-2-氧化物(324毫克，收率：85％)。MS(ESI):m/z＝279.0[M+H] +.
1-氯-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000022
向配有磁力搅拌器的50毫升茄形瓶中，加入氯氧磷(8.0毫升)然后缓慢加入7-氰基-6-(2-氟-6-甲基苯基)异喹啉-2-氧化物(324毫克，1.16毫摩尔)，然后将该反应液加热至回流搅拌16小时。反应经LCMS检测反应。反应结束后，反应液降至室温后，将反应液滴加到水(50毫升)中，用乙酸乙酯(20毫升×2)萃取。合并的有机相依次用水(20毫升)，饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，过滤，滤液经减压浓缩得粗品。该粗品经快速硅胶柱(石油醚:乙酸乙酯＝5:1)分离纯化得固体化合物1-氯-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(190毫克，收率：58％).MS(ESI):m/z＝297.0,299.0[M+H] +.
6-(2-氟-6-甲基苯基)-1-((4-甲氧苄基)氨基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000023
向1-氯-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(190毫克，0.64毫摩尔)的四氢呋喃(5毫克)中加入4-甲氧基苄胺(263毫克，1.92毫摩尔)和无水碳酸钠(339毫克，3.2毫摩尔)，将反应液加热至80度搅拌16小时。反应经LCMS检测。待反应液降至室温，减压浓缩溶剂得残留物。加入乙酸乙酯(20毫升)重新溶解该残留物，然后用水洗涤(20毫升)，分液，收集有机相，水相经乙酸乙酯(10毫升×2)萃取。合并有机相，然后依次用 水(20毫升)，饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，过滤。滤液经减压浓缩，得到粗品化合物6-(2-氟-6-甲基苯基)-1-((4-甲氧苄基)氨基)异喹啉-7-甲腈)(310毫克)。该粗品直接用于下一步反应。MS(ESI):m/z＝398.0[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000024
在室温下，向化合物6-(2-氟-6-甲基苯基)-1-((4-甲氧苄基)氨基)异喹啉-7-甲腈)(310毫克，0.64毫摩尔)的二氯甲烷(4.0毫升)溶液中，加入三氟乙酸(4.0毫升)，将反应液加热至60度搅拌16小时。反应经LCMS检测。待反应冷却至室温后，减压浓缩溶剂得残留物，然后向其中加入二氯甲烷(20毫升)。该溶液依次用饱和碳酸氢钠水溶液(20毫升)，饱和食盐水(20毫升)洗涤，无水硫酸钠干燥。过滤，滤液经减压浓缩得粗品，该粗品经快速硅胶柱(石油醚:乙酸乙酯＝5:1)分离纯化得固体化合物(石油醚/乙酸乙酯＝3/1-二氯甲烷/甲醇＝10/1)固体化合物1-氨基-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(140毫克，收率：78％)。MS(ESI):m/z＝278.0[M+H] +.
1H NMR(CDCl 3,400MHz)δ8.39(s,1H),8.12(t,J＝5.6Hz,1H),7.68(s,1H),7.39-7.33(m,1H),7.17(d,J＝7.6Hz,1H),7.09-7.05(m,2H),5.53(s,2H),2.18(s,3H).
1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000025
在冰浴冷却下，向1-氨基-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(200毫克，0.72毫摩尔)的二氯甲烷(4.0毫升)溶液中分批加入N-溴代琥珀酰亚胺(100毫克，0.72毫摩尔)。将反应液于0度搅拌0.5小时后，撤去冰浴，于室温继续搅拌0.5小时。反应经LCMS检测。加二氯甲烷(5.0毫升)稀释反应液，然后向其中加入10％的亚硫酸钠(5毫升)水溶液中，搅拌10分钟后，分液收集有机向。水相用二氯甲烷(5毫升×2)萃取。合并的有机相经水(10毫升)洗涤，无水硫酸钠干燥，过滤，滤液经减压浓缩得目标化合物1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(237毫克，收率：92％)。MS(ESI):m/z＝356.0,357.9[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)-4-(6-(4-甲基哌嗪-1-基)吡啶-3-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000026
在氩气保护下，向1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(80毫克，0.22毫摩尔)，(6-(4-甲基哌嗪-1-基)吡啶-3-基)硼酸(60毫克，0.27毫摩尔)，磷酸钾(95毫克， 0.44毫摩尔)和1,4-二氧六环(2.0毫升)/水(0.5毫升)的混合物中加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(18毫克,0.022毫摩尔)。将该反应液加热至100℃搅拌3小时。反应经LCMS检测。待反应液冷却至室温后，加乙酸乙酯(10毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩溶剂得粗品，该粗品经Prep-HPLC(NH 4HCO 3)分离得1-氨基-6-(2-氟-6-甲基苯基)-4-(6-(4-甲基哌嗪-1-基)吡啶-3-基)异喹啉-7-甲腈(37毫克，收率：36.4％)。MS(ESI):m/z＝453.1[M+H] +.
1H NMR(400MHz,CD 3OD)δ8.84(s,1H),8.10(d,J＝2.0Hz,1H),7.86(s,1H),7.61–7.54(m,2H),7.39–7.31(m,1H),7.15(d,J＝7.6Hz,1H),7.04(t,J＝8.8Hz,1H),6.90(d,J＝8.8Hz,1H),3.62–3.52(m,4H),2.61–2.50(m,4H),2.32(s,3H),2.09(s,3H).
实施例3:1-氨基-6-(2-氟-6-甲基苯基)-4-(1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000027
叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-1H-吡唑-1-基)哌啶-1-羧 酸酯
Figure PCTCN2021078966-appb-000028
在氩气保护下，向1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(60毫克，0.17毫摩尔)，叔-丁基4-(4-(4,4,5,5-四甲基-1,3,2-二噁硼戊环-2-基)-1H-吡唑-1-基)哌啶-1-羧酸酯(76毫克，0.20毫摩尔)，磷酸钾(72毫克，0.34毫摩尔)和1,4-二氧六环(2.0毫升)/水(0.5毫升)的混合物中加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(14毫克,0.017毫摩尔)。将该反应液加热至100℃搅拌10小时。反应经LCMS检测。待反应液冷却至室温后，加乙酸乙酯(5毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩溶剂得粗品，该粗品经Prep-HPLC(HCOOH)分离得叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-1H-吡唑-1-基)哌啶-1-羧酸酯(55毫克，收率：62％)。MS(ESI):m/z＝527.1[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)-4-(1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000029
在0度下，向叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-1H-吡唑-1-基)哌啶-1-羧酸酯(55毫克，0.104毫摩尔)的无水二氯甲烷(2.0毫升)中加入三氟乙酸 (1.0毫升)。该反应液于0度搅拌1小时。反应经LCMS检测。待反应结束后，反应溶液经减压浓缩溶液得油状残留物。该残留物经Prep-HPLC(NH 4HCO 3)分离得固体1-氨基-6-(2-氟-6-甲基苯基)-4-(1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7-甲腈(16毫克，收率：35.9％)。MS(ESI):m/z＝427.0[M+H] +.
1H NMR(400MHz,CD 3OD)δ8.85(s,1H),7.94(s,1H),7.88(s,1H),7.77(s,1H),7.62(s,1H),7.43–7.36(m,1H),7.20(d,J＝8.0Hz,1H),7.08(t,J＝8.8Hz,1H),4.36(s,1H),3.21(d,J＝12.8Hz,2H),2.79(m,2H),2.15(d,J＝8.0Hz,5H),1.98(m,2H).
实施例4:1-氨基-6-(2-氟-6-甲基苯基)-4-(2-(哌啶-4-基)噻唑-5-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000030
叔-丁基4-(5-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)噻唑-2-基)哌啶-1-羧酸酯
Figure PCTCN2021078966-appb-000031
在氩气保护下，向1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(60毫克，0.17毫摩尔)，叔-丁基4-(5-(4,4,5,5-四甲基-1,3,2-二噁硼戊环-2-基)噻唑-2-基)哌啶-1-羧酸酯(79毫克，0.20毫摩尔)，磷酸钾(72毫克，0.34毫摩尔)和1,4-二氧六环(1.2毫升)/水(0.4毫升)的混合物中加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(14毫克,0.017毫摩尔)。将该反应液加热至100℃搅拌10小时。反应经LCMS检测。待反应液冷却至室温后，加乙酸乙酯(5毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩得粗品。该粗品经Prep-HPLC(HCOOH)分离得叔-丁基4-(5-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)噻唑-2-基)哌啶-1-羧酸酯(22毫克，收率：22％)。MS(ESI):m/z＝544.1[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)-4-(2-(哌啶-4-基)噻唑-5-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000032
在0度下，向叔-丁基4-(5-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)噻唑-2-基)哌啶-1-羧酸酯(22毫克，0.041毫摩尔)的无水二氯甲烷(2.0毫升)中加入三氟乙酸(1.0毫升)。该反应液于0度搅拌1小时。反应经LCMS检测。待反应结束后，反应溶液经减压浓缩溶液得油状残留物。该残留物经Prep-HPLC(NH 4HCO 3)分离得固体1-氨基-6-(2-氟-6-甲基苯基)-4-(2-(哌啶-4-基)噻唑-5-基)异喹啉-7-甲腈(13毫克，收率：72.4％)。MS(ESI):m/z＝444.0[M+H] +.
1H NMR(400MHz,CD 3OD)δ8.87(s,1H),8.02(s,1H),7.73(s,1H),7.68(s,1H),7.39(m,1H),7.19(d,J＝7.6Hz,1H),7.07(t,J＝8.8Hz,1H),3.24(d,J＝12.0Hz,2H),2.87(t,J＝11.6Hz,2H),2.26–2.03(m,5H),1.84(m,2H).
实施例5:1-氨基-6-(2-氟-6-甲基苯基)-4-(5-(哌嗪-1-基)吡啶-2-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000033
叔-丁基4-(6-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)吡啶-3-基)哌嗪-1-羧酸酯
Figure PCTCN2021078966-appb-000034
在氩气保护下，向1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(60毫克，0.17毫摩尔)和叔-丁基4-(6-(三丁基甲锡烷基)吡啶-3-基)哌嗪-1-羧酸酯(186毫克，0.34毫摩尔)的无水甲苯溶液中，加入四(三苯基膦)钯(0)(38毫克，0.033毫摩尔)。将该反应液加热至120℃搅拌16小时。反应经LCMS检测。待反应液冷却至室温后，减压浓缩溶剂得粗品。该粗品经Prep-HPLC(HCOOH)分离得叔-丁基4-(6-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)吡啶-3-基)哌嗪-1-羧酸酯(50毫克，收率：54.3％)。MS(ESI):m/z＝539.1[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)-4-(5-(哌嗪-1-基)吡啶-2-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000035
在零度下，向叔-丁基4-(6-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)吡啶-3-基)哌嗪-1-羧酸酯(50毫克，0.092毫摩尔)的无水二氯甲烷(2.0毫升)中加入三氟乙酸(1.0毫升)。该反应液于0度搅拌1小时。反应经LCMS检测。待反应结束后，反应溶液经减压浓缩溶液得油状残留物。该残留物经Prep-HPLC(NH 4HCO 3)分离得固体1-氨基-6-(2-氟-6-甲基苯基)-4-(5-(哌嗪-1-基)吡啶-2-基)异喹啉-7-甲腈(24毫克，收率：59％)。MS(ESI):m/z＝439.1[M+H] +.
1H NMR(400MHz,MeOD)δ＝8.90(s,1H),8.39(d,J＝2.4Hz,1H),8.09(s,1H),8.02(s,1H),7.93(s,1H),7.61–7.56(m,1H),7.54(m,1H),7.41–7.35(m,1H),7.18(d,J＝7.6Hz,1H),7.07(d,J＝8.8Hz,1H),3.56–3.50(m,4H),3.41–3.35(m,4H),2.13(s,3H).
实施例6:1-氨基-6-(2-氟-6-甲基苯基)-4-(3-甲基-1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7- 甲腈
Figure PCTCN2021078966-appb-000036
叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-3-甲基-1H-吡唑-1-基)哌 啶-1-羧酸酯
Figure PCTCN2021078966-appb-000037
在氩气保护下，向1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(50毫克，0.14毫摩尔)，叔-丁基4-(3-甲基-4-(4,4,5,5-四甲基-1,3,2-二噁硼戊环-2-基)-1H-吡唑-1-基)哌啶-1-羧酸酯(66毫克，0.17毫摩尔)，磷酸钾(60毫克，0.28毫摩尔)和1,4-二氧六环(1.5毫升)/水(0.5毫升)的混合物中加入甲烷磺酸(2-二环己基膦基-2',4',6'-三-异丙基-1,1'-联苯基)(2'-氨基-1,1'-联苯-2-基)钯(II)(12毫克,0.014毫摩尔)。将该反应液加热至100℃搅拌3小时。反应经LCMS检测。待反应液冷却至室温后，加乙酸乙酯(5毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩得粗品。该粗品经Prep-HPLC(HCOOH)分离得叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-3-甲基-1H-吡唑-1-基)哌啶-1-羧酸酯(22毫克，收率：29.3％)。MS(ESI):m/z＝541.1[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)-4-(3-甲基-1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000038
在零度下，向叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-3-甲基-1H-吡唑-1-基)哌啶-1-羧酸酯(22毫克，0.041毫摩尔)的无水二氯甲烷(2.0毫升)中加入三氟乙酸(0.5毫升)。该反应液于室温搅拌2小时。反应经LCMS检测。待反应结束后，反应溶液经减压浓缩溶液得油状残留物。该残留物经Prep-HPLC(NH4HCO3)分离得固体1-氨基-6-(2-氟-6-甲基苯基)-4-(3-甲基-1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7-甲腈(12毫克，收率：67％)。MS(ESI):m/z＝441.1[M+H] +.
1H NMR(400MHz,MeOD)δ8.82(s,1H),7.81(s,1H),7.66(s,1H),7.39–7.32(m,2H),7.16(d,J＝7.6Hz,1H),7.04(t,J＝8.8Hz,1H),4.22(s,1H),3.14(d,J＝12.8Hz,2H),2.71(t,J＝11.6Hz,2H),2.05(m,8H),1.94–1.81(m,2H).
实施例7:1-氨基-6-(2-氟-6-甲基苯基)-4-(3-甲氧基-1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉- 7-甲腈
Figure PCTCN2021078966-appb-000039
叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-3-甲氧基-1H-吡唑-1-基) 哌啶-1-羧酸酯
Figure PCTCN2021078966-appb-000040
在氩气保护下，向1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(25毫克，0.07毫摩尔)，叔-丁基4-(3-甲氧基-4-(4,4,5,5-四甲基-1,3,2-二噁硼戊环-2-基)-1H-吡唑-1-基)哌啶-1-羧酸酯(28毫克，0.07毫摩尔)，磷酸钾(30毫克，0.14毫摩尔)和1,4-二氧六环(0.6毫升)/水(0.2毫升)的混合物中加入甲烷磺酸(2-二环己基膦基-2',4',6'-三-异丙基-1,1'-联苯基)(2'-氨基-1,1'-联苯-2-基)钯(II)(6.0毫克，0.007毫摩尔)。将该反应液加热至100℃搅拌3小时。反应经LCMS检测。待反应液冷却至室温后，加乙酸乙酯(5毫升×2)萃取。合并的有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩得粗品。该粗品经Prep-HPLC(HCOOH)分离得叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-3-甲氧基-1H-吡唑-1-基)哌啶-1-羧酸酯(10.3毫克，收率：26.3％)。MS(ESI):m/z＝557.1[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)-4-(3-甲氧基-1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000041
在零度下，向叔-丁基4-(4-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)-3-甲氧基-1H-吡唑-1-基)哌啶-1-羧酸酯(10.3毫克，0.018毫摩尔)的无水二氯甲烷(2.0毫升)中加入三氟乙酸(0.5毫升)。该反应液于室温搅拌2小时。反应经LCMS检测。待反应结束后，反应溶液经减压浓缩溶液得油状残留物。该残留物经Prep-HPLC(NH 4HCO 3)分离得固体1-氨基-6-(2-氟-6-甲基苯基)-4-(3-甲氧基-1-(哌啶-4-基)-1H-吡唑-4-基)异喹啉-7-甲腈(6.0毫克，收率：71.4％)。MS(ESI):m/z＝457.1[M+H] +.
1H NMR(400MHz,MeOD)δ8.80(s,1H),8.18–8.16(m,1H),7.86(s,1H),7.59(m,2H),7.38(d,J＝6.4Hz,1H),7.20(m,1H),7.08(d,J＝8.8Hz,1H),4.19–4.09(m,1H),3.75(d,J＝42.1Hz,3H),3.20(m,2H),2.75(m,2H),2.12(m,4H),1.97(s,3H).
实施例8:1-氨基-6-(2-氟-6-甲基苯基)-4-(6-(哌嗪-1-基)吡啶-2-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000042
叔-丁基4-(6-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)吡啶-2-基)哌嗪-1-羧酸酯
Figure PCTCN2021078966-appb-000043
在氩气保护下，向1-氨基-4-溴-6-(2-氟-6-甲基苯基)异喹啉-7-甲腈(60毫克，0.17毫摩尔)和叔-丁基4-(6-(三丁基甲锡烷基)吡啶-2-基)哌嗪-1-羧酸酯(186毫克，0.34毫摩尔)的无水甲苯溶液中，加入四(三苯基膦)钯(0)(38毫克，0.033毫摩尔)。将该反应液加热至120℃搅拌16小时。反应经LCMS检测。待反应液冷却至室温后，减压浓缩溶剂得粗品。该粗品经Prep-HPLC(HCOOH)分离得叔-丁基4-(6-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)吡啶-2-基)哌嗪-1-羧酸酯(51毫克，收率：56.2％))。MS(ESI):m/z＝539.1[M+H] +.
1-氨基-6-(2-氟-6-甲基苯基)-4-(6-(哌嗪-1-基)吡啶-2-基)异喹啉-7-甲腈
Figure PCTCN2021078966-appb-000044
在零度下，向叔-丁基4-(6-(1-氨基-7-氰基-6-(2-氟-6-甲基苯基)异喹啉-4-基)吡啶-2-基)哌嗪-1-羧酸酯(51毫克，0.092毫摩尔)的无水二氯甲烷(2.0毫升)中加入三氟乙酸(0.5毫升)。该反应液于室温搅拌2小时。反应经LCMS检测。待反应结束后，反应溶液经减压浓缩溶液得油状残留物。该残留物经Prep-HPLC(NH 4HCO 3)分离得固体1-氨基-6-(2-氟-6-甲基苯基)-4-(6-(哌嗪-1-基)吡啶-2-基)异喹啉-7-甲腈(28.5毫克，收率：70.0％)。MS(ESI):m/z＝439.1[M+H] +.
1H NMR(400MHz,MeOD)δ8.80(s,1H),8.43(s,1H),8.11(s,1H),7.60(m,1H),7.38–7.31(m,1H),7.15(d,J＝7.6Hz,1H),7.04(t,J＝8.8Hz,1H),6.89(d,J＝7.2Hz,1H),6.70(d,J＝8.4Hz,1H),3.51–3.42(m,4H),2.82–2.76(m,4H),2.08(s,3H).
生物测试例1 HPK1 ADP-Glo酶学活性测试
制备酶活测试缓冲液包含40mM Tris，pH7.5；20mM毫克Cl2；0.1毫克/毫升BSA；50μM DTT。化合物溶于100％DMSO，母液浓度为10mM。化合物的DMSO溶液从100uM起，连续做三倍梯度稀释，共十一个浓度，将稀释后的化合物用酶活测试缓冲液以1:20稀释好后，取1uL加入工作孔，每个浓度两复孔。阴性对照孔和阳性对照孔加入1:20稀释的DMSO溶液1uL。制备2.5×底物/ATP工作液为包含0.25ug/uLMBP蛋白和45uM ATP的酶活测试缓冲液，每个工作孔中加入2uL 2.5×底物/ATP工 作液。制备2.5×酶反应工作液为包含0.5ng/uL HPK1重组蛋白(Signalchem，货号M23-11G-10)的酶活测试缓冲液，每个工作孔中加入2uL 2.5×酶反应工作液，阴性对照孔仅加入2uL酶活测试缓冲液。贴好封板膜简单离心后置于室温反应30分钟，反应结束后每孔加入5uL ADP‐Glo试剂(Promega，货号V1901)室温反应40分钟，随后加入10uL激酶检测试剂(Promega，货号V1901)，室温反应20分钟后测定最终发光信号。
分别计算阳性孔和阴性孔的平均值，作为阳性对照值(Signal pos)和阴性对照值(Signal neg)。将工作孔信号值(Signal test)按公式Inhibition rate＝(Signal pos-Signal test)/(Signal pos–Signal neg)×100％计算抑制率。求得的抑制率在GraphPad Prism软件中按非线性拟合绘制浓度-抑制率曲线，计算IC 50。
生物测试例2 GLK ADP-Glo酶学活性测试
制备酶活测试缓冲液包含40mM Tris，pH7.5；20mM毫克Cl2；0.1毫克/毫升BSA；50μM DTT。化合物溶于100％DMSO，母液浓度为10mM。化合物的DMSO溶液从1mM起，连续做三倍梯度稀释，共十一个浓度，将稀释后的化合物用酶活测试缓冲液以1:20稀释好后，取1uL加入工作孔，每个浓度两复孔。阴性对照孔和阳性对照孔加入1:20稀释的DMSO溶液1uL。制备2.5×底物/ATP工作液为包含0.5ug/uL PKA底物多肽和105uM ATP的酶活测试缓冲液，每个工作孔中加入2uL 2.5×底物/ATP工作液。制备2.5×酶反应工作液为包含2.5ng/uL GLK重组蛋白(Signalchem，货号M25-11G-10)的酶活测试缓冲液，每个工作孔中加入2uL 2.5×酶反应工作液，阴性对照孔仅加入2uL酶活测试缓冲液。贴好封板膜简单离心后置于室温反应1小时，反应结束后每孔加入5uL ADP‐Glo试剂(Promega，货号V1901)室温反应40分钟，随后加入10uL激酶检测试剂(Promega，货号V1901)，室温反应20分钟后测定最终发光信号。
分别计算阳性孔和阴性孔的平均值，作为阳性对照值(Signal pos)和阴性对照值(Signal neg)。将工作孔信号值(Signal test)按公式Inhibition rate＝(Signal pos-Signal test)/(Signal pos–Signal neg)×100％计算抑制率。求得的抑制率在GraphPad Prism软件中按非线性拟合绘制浓度-抑制率曲线，计算IC 50。实验结果见下表：
表1 HPK1、GLK酶活实验结果
Figure PCTCN2021078966-appb-000045
其中，A表示IC 50值≤50nM；B表示50nM＜IC 50值≤500nM；C表示500nM＜IC 50值≤10uM。
生物测试例3 SLP76磷酸化细胞学测试
抑制HPK1可以抑制其下游SLP76的磷酸化。SLP76蛋白的磷酸化使用Jurkat(ATCC,Clone E6-1
Figure PCTCN2021078966-appb-000046
TIB-152 TM)细胞进行测试，实验的第一天将细胞用培养基(RPMI 1640+0.5％FBS)稀释到10 6/毫升，按每孔100uL，10 5细胞的量铺在96孔细胞培养板中，饥饿4小时培养。化合物溶于100％DMSO，母液浓度为4mM。化合物的DMSO溶液从10mM起，连续做四倍梯度稀释，共9个浓度，将4uL稀释后的化合物稀释到196uL 37度预热的RPMI 1640并混匀。取50uL最终稀释后的化合物加入细胞中，37度孵育20分钟，加入50uL稀释后的人CD3/CD28T细胞激活剂(Stemcell，货号：10971)，使激活剂的终浓度体积为总体系的1/40，37度孵育30分钟。反应结束后将细胞置于4度离心机1200rpm离心5分钟，吸走培养基，加入150uL细胞裂解液(ELISA kit中提供，Cell Signaling，货号30794C)在冰上放置30分钟使细胞充分裂解，裂解后的细胞吹打均匀后置于4度离心机以4000rpm离心5分钟，随后取50uL上清加入FastScan TM Phospho-SLP-76(Ser376)ELISA Kit(Cell Signaling，货号30794C)测试细胞SLP76的磷酸化水平。
分别计算阳性孔和阴性孔的平均值，作为阳性对照值(Signal pos)和阴性对照值(Signal neg)。将工作孔信号值(Signal test)按公式Inhibition rate＝(Signal pos-Signal test)/(Signal pos–Signal neg)×100％计算抑制率。求得的抑制率在GraphPad Prism软件中按非线性拟合绘制浓度-抑制率曲线，计算IC 50。实验结果见下表：
表2 SLP76磷酸化抑制实验结果
实施例编号 IC50(nM)
1 B
2 B
3 A
4 B
5 -
6 -
7 B
8 -
其中，A表示IC 50值≤1000nM；B表示1000nM＜IC 50值≤20000nM。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (13)

  1. 一种如下式I所示的化合物，或其药学上可接受的盐、光学异构体或水合物；
    Figure PCTCN2021078966-appb-100001
    其中，
    X选自N或CH，其中，所述的CH可以进一步被R 1取代；
    Ar环选自下组：6-10元芳香环，或5-10元的杂芳环；
    B环选自下组：6-12元芳香环，或5-12元的杂芳环；或含有选自下组的环结构单元的8-15元二环并环：6-10元芳香环、5-10元的杂芳环、3-10元杂环基、C3-C8环烷基；
    m为0、1、2或3；
    n和p各自独立地为0、1、2、3或4；
    Re选自下组：取代或未取代的C1-C6烷基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自N、S和O的杂原子的3-10元杂环基；其中，所述的取代优选包括一个或多个选自下组的基团：-N(Rc) 2
    Rc选自下组：H、或取代或未取代的C1-C5的烷基；
    Rf选自下组：氢、卤素、CN、取代或未取代的C1-C6烷基、-NH 2、R 9R 10N-C(＝O)-；
    各个R 1、R 2、R 3和Rg各自独立地选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-12元杂环基、取代或未取代的C6-C10芳基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-10元杂芳基、R 7-C(＝O)-、R 8-S(＝O) 2-、R 9R 10N-C(＝O)-、R 9R 10C(＝O)N(Rc)-、R 11R 12N-S(＝O) 2-；
    各个R 7、R 8、R 9、R 10、R 11、R 12各自独立地选自下组：H、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、取代或未取代的具有1-3个选自下组N、S和O的杂原子的3-12元杂环基；
    除非特别说明，所述的“取代”是指被选自下组的一个或多个(例如2个、3个、4个等)取代基所取代：卤素、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、甲基砜基、氧代(＝O)、-CN、羟基、羧基、C1-C6酰胺基(-C(＝O)-N(Rc) 2或-NH-C(＝O)(Rc))、-N(Rc) 2、或取代或未取代的选自下组的基团：C1-C6烷基、C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、-(CH 2)-C6-C10芳基、-(CH 2)-(具有1-3个选自N、S和O的杂原子的5-10元杂芳基)、-(具有1-3个选自N、S和O的杂原子的5-10元亚杂芳基)-(C1-C6烷基)、具有1-3个选自下组N、S和O的杂原子的3-12元杂环基(包括单环、螺环、桥环或并环)，且所述的取代基选自下组：卤素、C1-C6烷基、C1-C6亚烷基-OH、C1-C6烷氧基、氧代、-S(O) 2CH 3、-CN、-OH、C6-C10芳基、具有1-3个选自N、S和O的杂原子的3-10元杂芳基、-C(O)CHNH 2、-C(O)CHOH；
    且所述的式I化合物中，各个手性中心为R构型或S构型。
  2. 如权利要求1所述的化合物，或其药学上可接受的盐、光学异构体或水合物，其特征在于，所述的Re选自下组：取代或未取代的C1-C6烷基、取代或未取代的具有1-3个选自N、S和O的杂原子的3-10元杂环基。
  3. 如权利要求1所述的化合物，或其药学上可接受的盐、光学异构体或水合物，其特征在于，Ar环为5-7元的杂芳环；且所述的R 2选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基。
  4. 如权利要求1所述的化合物，或其药学上可接受的盐、光学异构体或水合物，其特征在于，所述的B环选自下组：苯环、含有苯环结构单元的8-15元二环并环、5-6元的杂芳环、含有5-6元杂芳环结构单元的8-15元二环并环；其中，所述的R 3选自下组：H、卤素、取代或未取代的C1-C6烷基、取代的-C(＝O)-；其中，所述的取代优选包括一个或多个选自下组的基团：-N(Rc) 2。
  5. 如权利要求1所述的化合物，或其药学上可接受的盐、光学异构体或水合物，其特征在于，所述的式I化合物具有如下式II所示的结构：
    Figure PCTCN2021078966-appb-100002
    其中，
    所述的Ra和Rb各自独立地选自下组：氢、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；
    所述的R 4选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、R 9R 10N-C(＝O)-；其中，所述的R 9、R 10选自下组：H、取代或未取代的C1-C6烷基；
    或所述的Rb和R 4与相连的环原子共同构成5-6元碳环或杂环，较佳地，所述的碳环或杂环为非芳香性环。
  6. 如权利要求5所述的化合物，或其药学上可接受的盐、光学异构体或水合物，其特征在于，所述的式I化合物具有如下式III所示的结构：
    Figure PCTCN2021078966-appb-100003
    其中，
    所述的Rd选自下组：H、卤素、CN、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、R 9R 10N-C(＝O)-；其中，所述的R 9、R 10选自下组：H、取代或未取代的C1-C6烷基。
  7. 如权利要求1所述的化合物，或其药学上可接受的盐、光学异构体或水合物，其特征在于，所述的式I化合物选自下组：
    Figure PCTCN2021078966-appb-100004
    Figure PCTCN2021078966-appb-100005
  8. 如权利要求1所述的化合物，或其药学上可接受的盐、光学异构体或水合物，其特征在于，所述的式I化合物选自下组：
    Figure PCTCN2021078966-appb-100006
    Figure PCTCN2021078966-appb-100007
  9. 一种药物组合物，其特征在于，所述的药物组合物包括(1)如权利要求1所述的化合物或其立体异构体或互变异构体，或其药学上可接受的盐、水合物或溶剂化物；(2)药学上可接受的载体。
  10. 如权利要求1所述的化合物或其立体异构体或互变异构体，或其药学上可接受的盐、水合物或溶剂化物，或如权利要求8所述的药物组合物的用途，其特征在于，用于制备预防和/或治疗与HPK1激酶的活性或表达量相关的疾病的药物组合物。
  11. 权利要求1所述的化合物及其药学上可接受的盐、立体异构体、酯、前药、溶剂化物和氘代化合物单独用药或者联合肿瘤化疗方案，其他肿瘤免疫治疗剂(小分子化合物及抗体包括但不限于PD-1、PD-L1、CTLA-4、TIM-3、TGF-β及其受体、LAG3拮抗剂或TLR4、TLR7、TLR8、TLR9、STING激动剂等)，放疗方案，肿瘤靶向药，肿瘤疫苗等，可在所述药剂之前、之后或同时施用，或者可以与其他已知疗法共施用。也可用作疫苗佐剂。
  12. 权利要求1所述的化合物及其药学上可接受的盐、立体异构体、酯、前药、溶剂化物和氘代化合物与CAR-T免疫疗法相结合在癌症免疫疗法中的应用。
  13. 权利要求10-12任一所述的应用，所述的疾病包括但不限于：淋巴瘤，母细胞瘤，髓母细胞瘤，视网膜母细胞瘤，肉瘤，脂肪肉瘤，滑膜细胞肉瘤，神经内分泌肿瘤，类癌肿瘤，胃泌素瘤，胰岛细胞癌，间皮瘤，神经鞘瘤，听神经瘤，脑膜瘤，腺癌，黑色素瘤，白血病或淋巴样恶性肿瘤，鳞状细胞癌，上皮鳞状细胞癌，肺癌，小细胞肺癌，非小细胞肺癌，腺癌肺癌，肺鳞癌，腹膜癌，肝细胞癌，胃癌，肠癌，胰腺癌，成胶质细胞瘤，子宫颈癌，卵巢癌，肝癌，膀胱癌，乳腺癌，转移性乳腺癌，结肠癌，直肠癌，结直肠癌，子宫癌，唾液腺癌，肾癌，前列腺癌，外阴癌，甲状腺癌，肛门癌，阴茎癌，梅克尔细胞癌，食管癌，胆道肿瘤，头颈部癌，血液恶性肿瘤，鼻咽癌，多发性骨髓瘤，大场绒毛腺瘤，非霍奇金淋巴瘤，骨癌，睾丸癌，霍奇金病，精元细胞瘤，口腔癌，脑癌，皮肤癌，乳腺导管癌，肾盂癌，肾母细胞瘤，食管腺瘤，视网膜细胞瘤，神经胶质瘤，神经纤维瘤，胃肠道间质瘤，原位癌，子宫内膜癌和骨髓增生异常综合征等。
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106817900A (zh) * 2014-07-17 2017-06-09 默克专利有限公司 新颖的萘啶和异喹啉及其作为cdk8/19抑制剂的用途
CN110678466A (zh) * 2017-03-30 2020-01-10 豪夫迈·罗氏有限公司 作为hpk1抑制剂的二氮杂萘类
CN110709392A (zh) * 2017-03-30 2020-01-17 豪夫迈·罗氏有限公司 作为hpk1抑制剂的异喹啉

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106817900A (zh) * 2014-07-17 2017-06-09 默克专利有限公司 新颖的萘啶和异喹啉及其作为cdk8/19抑制剂的用途
CN110678466A (zh) * 2017-03-30 2020-01-10 豪夫迈·罗氏有限公司 作为hpk1抑制剂的二氮杂萘类
CN110709392A (zh) * 2017-03-30 2020-01-17 豪夫迈·罗氏有限公司 作为hpk1抑制剂的异喹啉

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
AURÉLIE MALLINGER, SCHIEMANN KAI, RINK CHRISTIAN, SEJBERG JIMMY, HONEY MARK A., CZODROWSKI PAUL, STUBBS MARK, POESCHKE OLIVER, BUS: "2,8-Disubstituted-1,6-Naphthyridines and 4,6-Disubstituted-Isoquinolines with Potent, Selective Affinity for CDK8/19", ACS MEDICINAL CHEMISTRY LETTERS, VOL. 7, N. 6, 9 June 2016 (2016-06-09), pages 573 - 578, XP055581057, Retrieved from the Internet <URL:https://pubs.acs.org/doi/pdf/10.1021/acsmedchemlett.6b00022> [retrieved on 20190415], DOI: 10.1021/acsmedchemlett.6b00022 *

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