Deuterated hpk1 kinase inhibitors, methods of making and using the same

Abstract

The application provides a compound of a general formula I or a pharmaceutically acceptable salt, a stereoisomer, an ester, a prodrug, a solvate thereof, and a preparation method and application thereof, in particular, application of the compound of the general formula I or a pharmaceutically acceptable salt, a stereoisomer, an ester, a prodrug, a solvate thereof as an HPK1 kinase inhibitor in prevention and / or treatment of tumors, diseases caused by pathogen infection or related to pathogen infection. The compound of the general formula I contains at least one deuterium atom, has high stability in an animal body and good bioavailability.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a deuterated HPK1 kinase inhibitor, its preparation method, and its application. This invention also relates to the crystal form of the aforementioned deuterated HPK1 kinase inhibitor (particularly 4-(3-(((2-amino-5-(1-(1-trideuteratedmethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol and its salts), their preparation methods, and their applications. Background Technology

[0002] HPK1 kinase participates in many signaling cascades, including growth factor signaling, MAPK signaling, cytokine signaling, apoptosis signaling, and antigen receptor signaling. HPK1 kinase is a key functional activator of the JNK / SAPK signaling pathway. When activated, it can selectively activate the MAPK signaling pathway of C-Jun N-terminal kinase (JNK).

[0003] HPK1 kinase can serve as a target for immunotherapy. It is activated by lymphocyte antigen receptors and inhibits AP-1, which plays a role in tumor formation and development by promoting cell proliferation, inhibiting differentiation, and promoting tumor cell invasion and metastasis. Targeted disruption of HPK1 kinase alleles can increase the production of Th1 cytokines by T cells in the TCR response.

[0004] S Sawasdikosol (HPK1 as a novel target for cancer immunotherapy, Immunol Res, 54 (2012), pp. 262–265) reported that HPK1 kinase- / - T cells proliferate much faster than monocytic wild-type cells and resist prostaglandin E2 (PGE2)-mediated inhibition. Most notably, mice transfected with HPK1 kinase- / - T cells resisted tumor growth, and dendritic cells (DCs) lacking HPK1 kinase exhibited excellent antigenicity, making them a potential candidate for antitumor vaccines that enhance antitumor immune responses in HPK1 kinase- / - DCs. Eliminating the blockade of HPK1 kinase activity with small molecule inhibitors can activate the good antitumor activity of both cell types, ultimately synergistically amplifying their potential antitumor capabilities. Furthermore, the transfected HPK1 kinase was not expressed in major organs, suggesting that inhibitors of HPK1 kinase activity may not lead to any serious complications.

[0005] US2016158360A1 discloses a composition and method for enhancing immune responses and treating cancer, the composition comprising a PD-1 antagonist and an HPK1 antagonist, wherein the HPK1 antagonist comprises a compound that inhibits the serine / threonine kinase activity of HPK1.

[0006] It is evident that HPK1 kinase plays a crucial role in disease treatment, particularly cancer treatment, and the discovery of small molecule inhibitors of HPK1 kinase is currently in urgent need. Patent document CN110396087A discloses a heterocyclic compound that can serve as an HPK1 kinase inhibitor. Summary of the Invention

[0007] This invention provides a compound represented by general formula I:

[0008]

[0009] in:

[0010] A is selected from CR 10 Or N;

[0011] Q is selected from O or S;

[0012] x and z are independently selected from integers between 0 and 6 (e.g., 0, 1, 2, 3, 4, 5, 6);

[0013] y is 0 or 1;

[0014] Ar is selected from aromatic five-membered heterocyclic groups, aromatic six-membered heterocyclic groups, or phenyl groups. The aromatic five-membered heterocyclic group is selected from: furanyl, thiophene, pyrrole, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, or selenothiazolyl. The aromatic six-membered heterocyclic group is selected from: pyridinyl, pyridazinyl, pyrimidinyl, or pyrazinyl. Optionally, the H on the aromatic five-membered heterocyclic group, aromatic six-membered heterocyclic group, or phenyl group can be substituted by the following groups: -D, -SO2, -SO2N(C). 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl groups, halogens, -CN, -OCH2F, -OCHF2, -OCF3, C 1-10 Straight-chain / branched alkyl, -N(C)0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, C 3-10 Cycloalkyl, -O heterocycloalkyl, -N heterocycloalkyl, -N heterocyclic aromatic, -O heterocyclic aromatic, or -S heterocyclic aromatic, wherein the alkyl moiety may be substituted by one or more of the following groups: -SO2, -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl), halogen, -CN, -OCH2F, -OCHF2, -OCF3, -N(C 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, -CO(C) 0-10 Alkyl), -COO(C 0-10 Alkyl), -N heterocyclic aromatic, -O heterocyclic aromatic or -S heterocyclic aromatic; wherein one or more H atoms attached to the C atom or heteroatom can be substituted with deuterium;

[0015] R2 is selected from: -H, -D, halogen, -NO2, -CN, C 1-10 Straight-chain / branched alkyl, C 3-10 cycloalkyl, -N(C) 0-10 Alkyl)(C 0-10 Alkyl groups, -CF3, -OCF3, -OCHF2, -OCH2F, or -OC 0-10 Alkyl group; wherein one or more H atoms bonded to the C atom can be substituted with deuterium;

[0016] B1, B2, B3, B4 and B5 are independently selected from C or N (when B1, B2, B3, B4 or B5 is N, the corresponding R3, R4, R5, R6 and R7 do not exist);

[0017] When present, R3, R4, R5, R6, and R7 are independently selected from: -H, -D, halogen, -CN, and -OC. 0-10 Alkyl, -CO(C) 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10Alkyl), C 1-10 Straight-chain / branched alkyl groups, heteroalkyl groups containing O or N, -N(C) 0-10 Alkyl)(C 0-10 Alkyl), C 3-10 cycloalkyl, -C≡CR 10 -O heterocyclic alkyl, -N heterocyclic alkyl, or R5 and R4, R4 and R3, R3 and R7, R7 and R6 form C with the carbon atoms between them. 3-8 cycloalkyl or C containing -O-, -S- 3-8 Heterocyclic alkyl, -N-heterocyclic aromatic, -O-heterocyclic aromatic or -S-heterocyclic aromatic, phenyl, wherein the alkyl moiety may be substituted by one or more of the following groups: -SO2, -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl groups, halogens, -CN, -OCH2F, -OCHF2, -OCF3, C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, C 3-10 Cycloalkyl, -O heterocycloalkyl, -N heterocycloalkyl, -N heterocyclic aromatic, -O heterocyclic aromatic or -S heterocyclic aromatic; wherein one or more H atoms attached to the C atom or heteroatom can be substituted with deuterium;

[0018] R8 and R9 are independently selected from: -H, -D, halogen, C 1-10 Straight-chain / branched alkyl; wherein one or more H atoms bonded to the C atom can be substituted with deuterium;

[0019] R 10 Selected from: H, -D, C 1-5 Straight-chain / branched alkyl, C 3-10 cycloalkyl, In this case, one or more H atoms bonded to the C atom can be replaced by deuterium;

[0020] R 11 R 12Independently selected from: -H, -D, -CF3, -CHF2H, -CH2F, C 1-10 Straight-chain / branched alkyl, -CH=C(C 0-10 Alkyl)(C 0-10 Alkyl), -C≡C(C 0-10 Alkyl), C 3-10 Cycloalkyl, aromatic five-membered ring or aromatic six-membered ring, or R 11 R 12 With R 11 and R 12 The carbon atoms between them form C 3-8 cycloalkyl or C containing -O-, -S- 3-8 Heterocyclic alkyl, C 4-9 Polycyclic alkyl, C 5-10 Spirocycloalkyl, C 4-9 Bridged cycloalkyl, C 3-7 cyclic lactam, C 3-7 cyclic lactones, C 3-7 Cyclic ketones, wherein the alkyl moiety may be substituted with one or more of the following groups: -SO2, -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl), halogen, -CN, -OCH2F, -OCHF2, -OCF3, -N(C 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, -N heterocyclic aromatic, -O heterocyclic aromatic, or -S heterocyclic aromatic; wherein one or more H atoms attached to the C atom or heteroatom may be substituted with deuterium;

[0021] Furthermore, compounds of general formula I contain at least one deuterium atom.

[0022] In one embodiment of the present invention, A is CR. 10 Especially CH.

[0023] In one embodiment of the present invention, Q is 0.

[0024] In one embodiment of the present invention, x is 0.

[0025] In one embodiment of the present invention, z is 1.

[0026] In one embodiment of the present invention, y is 1.

[0027] In one embodiment of the present invention, B1, B2, B3, B4, and B5 are all C, that is, in general formula I, for

[0028] In another embodiment of the invention, at least one of B1, B2, B3, B4 and B5 is N.

[0029] Specifically, B2 is C, and at least one of B1, B3, B4, and B5 is N.

[0030] More specifically, B2 is C, and B1 is N.

[0031] More specifically, B2 is C, and B3 is N.

[0032] More specifically, B2 is C, and B4 is N.

[0033] More specifically, B2 is C, and B5 is N.

[0034] More specifically, B2 is C, and B3 and B4 are N, or both B3 and B5 are N.

[0035] Specifically, Ar is selected from: thiazolyl, selenothiazolyl, imidazolel, pyrazolyl and pyridinyl.

[0036] In one embodiment of the present invention, the compound of general formula I has the following structure:

[0037]

[0038] Among them, ring E is selected from:

[0039] In ring E, each R0 is independently selected from: -H, -D, C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, -CO(C) 0-10 alkyl) or C 3-10 Cycloalkyl; wherein the H atom attached to the C atom or heteroatom may be replaced by deuterium;

[0040] R1 is selected from: -H, -D, -O heterocyclic alkyl, -N heterocyclic alkyl, C 1-10 Straight-chain / branched alkyl, C 3-10 cycloalkyl, -OC 0-10 Alkyl, -N(C)0-10 Alkyl)(C 0-10 alkyl), -SO2(C 0-10 Alkyl), -CO(C) 0-10 alkyl), -O-phenyl, -S(C 0-10 Alkyl), -N heterocyclic aromatic group, -O heterocyclic aromatic group or -S heterocyclic aromatic group; wherein the H atom attached to the C atom or heteroatom can be substituted with deuterium;

[0041] R 2-9 It has the corresponding definitions of the present invention above;

[0042] Furthermore, R 0-9 It contains at least one deuterium atom.

[0043] In one embodiment of the present invention, in general formula II, R1 contains at least one deuterium atom; more specifically, for example, R1 contains at least one deuterium atom, while R... 2-9 It does not contain deuterium atoms.

[0044] In another embodiment of the present invention, in general formula II, R2 contains at least one deuterium atom.

[0045] In another embodiment of the present invention, in general formula II, R3 contains at least one deuterium atom.

[0046] In another embodiment of the present invention, in general formula II, R4 contains at least one deuterium atom.

[0047] In another embodiment of the present invention, in general formula II, R5 contains at least one deuterium atom.

[0048] In another embodiment of the present invention, in general formula II, R6 contains at least one deuterium atom.

[0049] In another embodiment of the present invention, in general formula II, R8 and / or R9 contain at least one deuterium atom.

[0050] Specifically, each R0 is independently selected from: C 1-5 Straight-chain / branched alkyl or -N(C) 0-10 Alkyl)(C 0-10 Alkyl group, wherein the H atom attached to the C atom can be replaced by deuterium.

[0051] More specifically, each R0 is independently selected from: -H, -D, -CH3, -CH2CH3 or -NH2.

[0052] Specifically, R1 is selected from: -O heterocyclic alkyl or -N heterocyclic alkyl, -SO2(C 0-3 alkyl), -O-phenyl, -S(C 0-4 Alkyl), C3-6 cycloalkyl or C 3-5 Straight-chain / branched alkyl groups, wherein the H atom attached to the C atom or heteroatom can be replaced by deuterium.

[0053] More specifically, R1 is selected from: -CH3、 The H atoms bonded to the C or N atoms can be replaced by deuterium.

[0054] More specifically, R1 is selected from: -CH2D, -CHD2, -CD3

[0055] Specifically, when R0 and R1 are adjacent, R0 and R1 form C with the carbon atoms between them. 3-8 cycloalkyl or C containing -O-, -S- 3-8 Heterocyclic alkyl, -N heterocyclic aromatic, -O heterocyclic aromatic or -S heterocyclic aromatic, phenyl.

[0056] Specifically, R2 is selected from: -H, -D, halogen, -NO2, -CN, C 1-5 Straight-chain / branched alkyl, C 3-10 cycloalkyl, -N(C) 0-10 Alkyl)(C 0-10 Alkyl groups, -CF3, -OCF3, -OCHF2, -OCH2F, or -OC 0-10 Alkyl groups, wherein the H atoms attached to the C or N atoms can be replaced by deuterium.

[0057] More specifically, R2 is selected from: -NO2, -N(C 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl groups and -OCF3, wherein the H atom attached to the C or N atom can be replaced by deuterium.

[0058] More specifically, R2 is selected from: -NH2, -NHD, -ND2, or -NO2.

[0059] Specifically, R3 is selected from: -H, -D, halogen, -OC. 0-10 Alkyl, -CO(C) 0-10 Alkyl), C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl) or C 3-10 Cycloalkyl, wherein the H atom attached to the C atom can be replaced by deuterium.

[0060] More specifically, R3 is selected from: -H, -D, halogen, -OC 0-10 Alkyl, C 1-10 Straight-chain / branched alkyl groups, wherein the H atom attached to the C atom can be replaced by deuterium.

[0061] More specifically, R3 is selected from: -H, -D, -F, -OCH3, -OCH2D, -OCHD2, -OCD3.

[0062] Specifically, R4 is selected from: -H, -D, halogen, -OC. 0-10 Alkyl, -CO(C) 0-10 Alkyl), -CN, C 3-10 cycloalkyl, -C≡CR 10 C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10 Alkyl), -O heterocyclic alkyl, or -N heterocyclic alkyl, wherein the H atom attached to the C or N atom can be replaced by deuterium.

[0063] More specifically, R4 is selected from: -H, -D, halogen, -OC 0-10 Alkyl, -CN, C 3-10 cycloalkyl or -C≡CR 10 In this embodiment, the H atom bonded to the C atom can be replaced by deuterium. In one embodiment of the invention, R... 10 The H atom attached to the C atom can be replaced by deuterium.

[0064] In one embodiment of the present invention, R4 is selected from: -H, -D, -F, -Cl, -OCH3, -OCH2D, -OCHD2, -OCD3, -CN, or -C≡CR 10 .

[0065] Specifically, R5, R6, and R7 are independently selected from: -H, -D, halogen, -CN, and -OC. 0-10 Alkyl, -CO(C) 0-10 Alkyl), C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10 Alkyl), C 3-10 cycloalkyl, -C≡CR 10 -O heterocyclic alkyl or -N heterocyclic alkyl, C containing O or N 1-5 Straight-chain / branched alkyl groups, or carbon atoms between R6, R7 and R6 and R7 forming C 3-8 cycloalkyl or C containing -O-, -S- 3-8 Heterocyclic alkyl groups, wherein the H atom attached to the C atom or heteroatom can be replaced by deuterium.

[0066] More specifically, R5, R6, and R7 are independently selected from: -H, -D, halogen, -CN, and C. 1-3 Straight-chain / branched alkyl, -OC 0-3 Alkyl, -CO(C) 0-3 Alkyl groups, N-containing C 1-3 Straight-chain / branched alkyl groups, or carbon atoms between R6, R7 and R6 and R7 forming C 3-8 cycloalkyl or C containing -O- 3-8 Heterocyclic alkyl groups, wherein the H atom attached to the C or N atom can be replaced by deuterium.

[0067] More specifically, R5, R6, and R7 are independently selected from: -H, -D, -F, -Cl, -CH3, -CH2NH2, -CH2NH(CH3), -CH2N(CH3)2, -CN, -OCH3, -COCH3, or R6, R7 and the carbon atoms between R6 and R7 form a -O-containing five-membered cycloalkyl group, wherein the H atom attached to the C or N atom can be replaced by deuterium.

[0068] In one embodiment of the present invention, R5 is selected from: -H, -D, -F, -Cl, -CH3, -CH2D, -CHD2, -CD3, -OCH3, -COCH3, -CH2NH2, -CH2N(CH3)2, -CN, -OCH2D, -OCHD2, -OCD3, -COCD3, -CH2N(CD3)2, -CH2N(CH3)(CD3).

[0069] In one embodiment of the present invention, R6 is selected from: -H, -D, -F, -Cl, -CH3, -CH2D, -CHD2, -CD3, -OCH3, -COCH3, -CH2NH2, -CH2N(CH3)2, -CN, -OCH2D, -OCHD2, -OCD3, -COCD3, -CH2N(CD3)2, -CH2N(CH3)(CD3).

[0070] In one embodiment of the present invention, R7 is selected from: -H, -D, -F, -Cl, -CH3, -CH2D, -CHD2, -CD3, -OCH3, -COCH3, -CH2NH2, -CH2N(CH3)2, -CN, -OCH2D, -OCHD2, -OCD3, -COCD3, -CH2N(CD3)2, -CH2N(CH3)(CD3).

[0071] In one embodiment of the present invention, R 10 for In one embodiment of the present invention, R 10The H atom attached to the C atom can be replaced by deuterium.

[0072] Specifically, R 11 and R 12 Independently selected from: -H, -D, -CF3, -CHF2H, -CH2F, C 1-10 Straight-chain / branched alkyl, -CH=C(C 0-10 Alkyl)(C 0-10 Alkyl), C 3-10 Cycloalkyl or aromatic six-membered ring groups, or R 11 R 12 With R 11 and R 12 The carbon atoms between them form C 3-8 cycloalkyl, C 4-7 fused cycloalkyl, C 5-9 Spirocycloalkyl, C 4-9 Bridged cycloalkyl, C 3-7 cyclic lactam, C 3-7 cyclic lactones, C 3-7 Cyclic ketones, in which the H atom on the C atom can be substituted with the following groups: -SO2, -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl groups, halogens, -CN, -OCH2F, -OCHF2, -OCF3, C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, -CO(C) 0-10 Alkyl), C 3-10 Cycloalkyl, -O heterocycloalkyl, -N heterocycloalkyl, -N heterocyclic aromatic, -O heterocyclic aromatic, or -S heterocyclic aromatic, wherein the alkyl moiety may be substituted by one or more of the following groups: -SO2, -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl), halogen, -CN, -OCH2F, -OCHF2, -OCF3, -N(C 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, -CO(C) 0-10 Alkyl), -N heterocyclic aromatic, -O heterocyclic aromatic or -S heterocyclic aromatic; wherein the H atom attached to the C atom or heteroatom can be replaced by deuterium.

[0073] More specifically, R 11 and R 12 Independently selected from: -H, -D, -CF3, -CHF2, -CH2F, C 1-5 Straight-chain / branched alkyl groups, -CH=CH(C) 0-10 Alkyl), C 3-10 Cycloalkyl or aromatic six-membered ring groups, or R 11 R 12 With R 11 and R 12 The carbon atoms between them form C 3-6 cycloalkyl, C 4-6 Polycyclic alkyl, C 5-8 Spirocycloalkyl, C 4-8 Bridged cycloalkyl, C 3-7 cyclic lactam, C 3-7 cyclic lactones, C 3-7 Cyclic ketones, wherein the alkyl moiety can be substituted with the following groups: -SO2, -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl groups, halogens, -CN, -OCH2F, -OCHF2, -OCF3, C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10alkyl), -OC 0-10 Alkyl, -CO(C) 0-10 Alkyl), C 3-10 Cycloalkyl, -O heterocycloalkyl, -N heterocycloalkyl, -N heterocyclic aromatic, -O heterocyclic aromatic, or -S heterocyclic aromatic, wherein the alkyl moiety may be substituted by one or more of the following groups: -SO2, -SO2N(C 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)SO2(C 0-10 Alkyl), -CON(C) 0-10 Alkyl)(C 0-10 alkyl), -N(C) 0-10 Alkyl)CO(C 0-10 alkyl), -N(C) 0-10 Alkyl)COO(C 0-10 Alkyl), -OCON(C) 0-10 Alkyl)(C 0-10 Alkyl), halogen, -CN, -OCH2F, -OCHF2, -OCF3, -N(C 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl, -CO(C) 0-10 Alkyl), -N heterocyclic aromatic, -O heterocyclic aromatic or -S heterocyclic aromatic; wherein the H atom attached to the C atom or heteroatom can be replaced by deuterium.

[0074] More specifically, R 11 and R 12 Independently selected from: -H, -D, -CF3, -CHF2, -CH2F, -CH3, -CH2CH3, -CH=CH2, or R 11 R 12 With R 11 and R 12 The formation of carbon atoms between In this case, the H atoms bonded to the C or N atoms can be replaced by deuterium.

[0075] More specifically, R 11 and R 12 Independently selected from: -H, -D, -CF3, -CHF2, -CDF2, -CH2F, -CD2F, -CH3, -CH2D, -CHD2, -CD3, -CH2CH3, -CH2CD3,

[0076] Specifically, R8 and R9 are independently selected from: -H, -D, and -C. 1-10 Straight-chain / branched alkyl groups, wherein the H atom attached to the C atom can be replaced by deuterium.

[0077] More specifically, R8 and R9 are independently selected from: -H, -D, and -C. 1-3 Straight-chain / branched alkyl groups, wherein the H atom attached to the C atom can be replaced by deuterium.

[0078] More specifically, R8 and R9 are independently selected from: -H, -D, -CH3, -CH2D, -CHD2, -CD3.

[0079] Specifically, compounds of general formula I have the following structures: (1)

[0081]

[0082]

[0083]

[0084]

[0085]

[0086] (2)

[0088]

[0089]

[0090]

[0091]

[0092]

[0093] (3)

[0095]

[0096]

[0097]

[0098]

[0099]

[0100] (4)

[0102]

[0103]

[0104]

[0105]

[0106] (5)

[0108]

[0109]

[0110]

[0111]

[0112] Specifically, in the above compounds, any atom not specified as deuterium exists at its natural isotopic abundance.

[0113] Specifically, in the above compounds, the position designated as "deuterium" has at least (for example) 95% deuterium incorporation.

[0114] This invention provides a method for preparing a compound of general formula I, comprising the following steps:

[0115] (1) and A condensation reaction occurs to produce R 13 Selected from: halogen or R 14 Selected from: -OH or -F;

[0116] (2) with Ar-R 15 A condensation reaction occurs to produce R 15 Selected from: -Br ​​or -SnBu3.

[0117] Specifically, in step (1) above, R 13 It is -Br.

[0118] Specifically, in step (2) above, R 13 For -Br or When R 13 When it is -Br, R 15 For -SnBu3, when R13 yes At that time, R 15 It is -Br.

[0119] The present invention also provides pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, and solvates of compounds of the above general formula I.

[0120] Specifically, the pharmaceutically acceptable salts mentioned above include acid addition salts and base addition salts.

[0121] Specifically, the aforementioned acid addition salts include, but are not limited to, salts from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphonic acid, as well as salts from organic acids such as aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkanic acids, hydroxyalkanic acids, alkanedioic acids, aromatic acids, and aliphatic and aromatic sulfonic acids. Therefore, these salts include, but are not limited to, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, hydrochlorides, hydrobromates, iodates, acetates, propionates, octanoates, isobutyrates, oxalates, malonates, succinates, octanoates, sebacic acid salts, fumarates, maleates, amygdalinates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, maleates, tartrates, and methanesulfonates, as well as salts of amino acids such as arginine salts, gluconates, and galacturonic acids. Acid addition salts can be prepared by contacting a sufficient amount of the desired acid in a conventional manner to form a salt. The free base can be regenerated by contacting the salt with a base and then separated in a conventional manner.

[0122] Specifically, the aforementioned base addition salts form with metals or amines, such as hydroxides of alkali metals and alkaline earth metals, or with organic amines. Examples of metals used as cations include, but are not limited to, sodium, potassium, magnesium, and calcium. Suitable amines include, but are not limited to, N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine (ethane-1,2-diamine), N-methylglucosamine, and procaine. Base addition salts can be prepared by contacting the free acid form with a sufficient amount of the desired base in a conventional manner to form a salt. The free acid form can be regenerated by contacting the salt form with an acid, and the free acid can be separated in a conventional manner.

[0123] In one embodiment of the present invention, the pharmaceutically acceptable salt is a hydrochloride salt.

[0124] Specifically, the aforementioned stereoisomers include enantiomers, diastereomers, and geometric isomers. Some compounds of the present invention have cycloalkyl groups that can be substituted at more than one carbon atom; in this case, all their geometric forms, including cis and trans, and mixtures thereof, are within the scope of the present invention.

[0125] Specifically, the solvates mentioned above refer to the physical combination of the compounds of the present invention with one or more solvent molecules. This physical combination includes various degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, solvates can be isolated, for example when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" includes both solution-phase and separable solvates. Representative solvates include ethanolides, methanolides, etc. A "hydrate" is a solvate in which one or more solvent molecules are H₂O.

[0126] Specifically, the aforementioned prodrug refers to a compound of formula I that is suitable for administration to patients without excessive toxicity, irritation, or allergic reactions, and is effective for its intended purpose, including acetals, esters, and zwitterionic forms. The prodrug is converted in vivo (e.g., through hydrolysis in the blood) to yield the parent compound of the above formula.

[0127] The present invention also provides a compound of the above general formula I and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates.

[0128] Specifically, the present invention provides a crystal form of 4-(3-(((2-amino-5-(1-(1-trideuterated methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yne-2-ol (which has the following structure).

[0129]

[0130] Specifically, the crystal form mentioned above is crystal form A, whose XRPD spectrum has characteristic peaks (main characteristic diffraction peaks) at at least three (or all) positions among the positions with 2θ values ​​of 13.1°±0.2°, 16.3°±0.2°, 17.5°±0.2°, and 23.8°±0.2°.

[0131] Specifically, the XRPD pattern of crystal form A also has characteristic peaks (minor characteristic diffraction peaks) at at least three (at least four, at least five, at least six, at least seven, or all) positions with 2θ values ​​of 8.1°±0.2°, 12.2°±0.2°, 15.3°±0.2°, 18.0°±0.2°, 19.3°±0.2°, 19.5°±0.2°, 21.3°±0.2°, and 21.6°±0.2°.

[0132] Specifically, the crystal form A described above has essentially the following characteristics: Figure 1 The XRPD map shown.

[0133] Specifically, the DSC spectrum of crystal form A above has an endothermic peak at approximately 168.8 °C.

[0134] Specifically, the crystal form A described above has essentially the following characteristics: Figure 2 The DSC spectrum shown.

[0135] Specifically, the weight loss of crystal form A when heated from room temperature to 170°C is approximately 1.1%.

[0136] Specifically, the crystal form A described above has essentially the following characteristics: Figure 2 The TGA spectrum shown.

[0137] Specifically, crystal form A is the amorphous form.

[0138] Specifically, the crystal form mentioned above is crystal form B, whose XRPD spectrum has characteristic peaks (main characteristic diffraction peaks) at at least three (or all) of the positions with 2θ values ​​of 5.7°±0.2°, 11.3°±0.2°, 22.7°±0.2°, and 23.5°±0.2°.

[0139] Specifically, the XRPD pattern of crystal form B also has characteristic peaks (minor characteristic diffraction peaks) at at least three (at least four, at least five, or all) of the positions with 2θ values ​​of 7.1°±0.2°, 8.8°±0.2°, 14.1°±0.2°, 17.0°±0.2°, 18.0°±0.2°, and 18.8°±0.2°.

[0140] Specifically, the aforementioned crystal form B has essentially the following properties: Figure 6 The XRPD map shown.

[0141] Specifically, the DSC spectrum of the aforementioned crystal form B has an endothermic peak at at least one of approximately 59.5℃, 95.6℃, 150.8℃, and 160.9℃.

[0142] Specifically, the aforementioned crystal form B has essentially the following properties: Figure 7 The DSC spectrum shown.

[0143] Specifically, the weight loss of crystal form B when heated from room temperature to 70°C is approximately 13.2%, and the weight loss when heated further to 170°C is approximately 8.5%.

[0144] Specifically, the aforementioned crystal form B has essentially the following properties: Figure 7 The TGA spectrum shown.

[0145] Specifically, the crystal form B mentioned above is an EtOAc solvate.

[0146] Specifically, the present invention also provides the crystal form of 4-(3-(((2-amino-5-(1-(1-trideuterated methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yne-2-ol hydrochloride.

[0147] Specifically, the crystal form mentioned above is crystal form A, whose XRPD spectrum has characteristic peaks (main characteristic diffraction peaks) at at least three (at least four, or all) positions among the positions with 2θ values ​​of 13.0°±0.2°, 16.3°±0.2°, 17.5°±0.2°, 19.4°±0.2°, and 23.8°±0.2°.

[0148] Specifically, the XRPD pattern of crystal form A also has characteristic peaks (minor characteristic diffraction peaks) at at least three (at least four, or all) of the positions with 2θ values ​​of 8.1°±0.2°, 12.1°±0.2°, 15.3°±0.2°, 18.0°±0.2°, and 21.4°±0.2°.

[0149] Specifically, the crystal form A described above has essentially the following characteristics: Figure 11 The XRPD map shown.

[0150] Specifically, the DSC spectrum of crystal form A has endothermic peaks at approximately 81.9 °C and approximately 156.0 °C.

[0151] Specifically, the crystal form A described above has essentially the following characteristics: Figure 12 The DSC spectrum shown.

[0152] Specifically, the weight loss of crystal form A when heated from room temperature to 150°C is approximately 8.6%.

[0153] Specifically, the crystal form A described above has essentially the following characteristics: Figure 12 The TGA spectrum shown.

[0154] The present invention also provides a method for preparing the above-mentioned crystal form.

[0155] Specifically, the above preparation method is selected from one or more combinations of antisolvent addition, anti-antisolvent addition, gas-solid diffusion, room temperature suspension stirring, 5°C suspension stirring, slow evaporation, slow cooling, gas-liquid diffusion, and polymer induction.

[0156] Specifically, the above-mentioned antisolvent addition method includes: dissolving the target product in a good solvent, and then adding an antisolvent to the resulting solution (and then, for example, evaporating at room temperature to obtain a solid, or stirring at -20°C to obtain a solid).

[0157] Specifically, the above-mentioned anti-antisolvent addition method includes: dissolving the target product in a good solvent, and then adding the resulting solution to an antisolvent (and then, for example, evaporating at room temperature to obtain a solid, or stirring at -20°C to obtain a solid).

[0158] Specifically, the above-mentioned gas-solid diffusion method, room temperature suspension stirring method, 5°C suspension stirring method, slow evaporation method, slow cooling method, and gas-liquid diffusion method all include: dissolving the target product in a solvent and then drying it to obtain a solid.

[0159] Specifically, the above-mentioned polymer-induced method includes: dissolving the target product in a solvent, adding a polymer, and evaporating it at room temperature to obtain a solid.

[0160] Specifically, the preparation method of crystal form A of the above-mentioned 4-(3-(((2-amino-5-(1-(1-trideuterated methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol is selected from one or more combinations of the following: antisolvent addition, anti-antisolvent addition, gas-solid diffusion, room temperature suspension stirring, 5°C suspension stirring, slow evaporation, slow cooling, gas-liquid diffusion, and polymer induction.

[0161] Specifically, for the preparation of the above-mentioned crystal form A, for the antisolvent addition method, the good solvent can be selected from: MeOH, acetone, DMSO, EtOAc, EtOH, DCM, CHCl3, THF, IPA, ACN, 1,4-dioxane, and the antisolvent can be selected from: MTBE, toluene, n-heptane, and water.

[0162] In one embodiment of the present invention, in the antisolvent addition method, the good solvent can be selected from: MeOH, acetone, DMSO, and the antisolvent is MTBE.

[0163] In another embodiment of the present invention, in the antisolvent addition method, the good solvent can be selected from: EtOH, DCM, and the antisolvent is toluene.

[0164] In another embodiment of the present invention, in the antisolvent addition method, the good solvent can be selected from: CHCl3, THF, IPA, and the antisolvent is n-heptane.

[0165] In another embodiment of the present invention, in the antisolvent addition method, the good solvent may be selected from: acetone, ACN, 1,4-dioxane, and the antisolvent is water.

[0166] Specifically, for the preparation of the above-mentioned crystal form A, for the anti-antisolvent addition method, the good solvent can be selected from: MeOH, MIBK, acetone, anisole, EtOH, THF, EtOAc, DCM, and the antisolvent can be selected from: toluene, n-heptane, water, MTBE.

[0167] In one embodiment of the present invention, in the anti-antisolvent addition method, the good solvent can be MIBK and the antisolvent can be toluene.

[0168] In another embodiment of the present invention, in the anti-antisolvent addition method, the good solvent can be selected from acetone and anisole, and the antisolvent can be n-heptane.

[0169] In another embodiment of the present invention, in the anti-antisolvent addition method, the good solvent can be selected from EtOH and THF, and the antisolvent can be water.

[0170] In another embodiment of the present invention, in the anti-antisolvent addition method, the good solvent can be selected from EtOAc and DCM, and the antisolvent can be MTBE.

[0171] Specifically, for the preparation of the above-mentioned crystal form A, for the gas-solid diffusion method, the solvent can be selected from: water, DCM, EtOH, MeOH, ACN, THF, CHCl3, acetone, DMSO, EtOAc, 1,4-dioxane, IPA.

[0172] Specifically, for the preparation of the above-mentioned crystal form A, for the room temperature suspension stirring method, the solvent can be selected from: MTBE, IPAc, n-heptane, toluene, water, EtOH / toluene (e.g., in a 1:3 ratio, v / v), DMSO / MTBE (e.g., in a 1:4 ratio, v / v), acetone / water (e.g., in a 1:4 ratio, v / v), IPA / n-heptane (e.g., in a 1:4 ratio, v / v), EtOAc / n-heptane (e.g., in a 1:4 ratio, v / v), anisole / toluene (e.g., in a 1:4 ratio, v / v), DMAc / water (e.g., in a 1:4 ratio, v / v), THF / water (e.g., in a 1:4 ratio, v / v).

[0173] Specifically, for the preparation of the above-mentioned crystal form A, for the 5°C suspension stirring method, the solvent can be selected from: MTBE, toluene, water, IPA / n-heptane (e.g., in a 1:2 ratio, v / v), MEK / n-heptane (e.g., in a 1:2 ratio, v / v), EtOAc / toluene (e.g., in a 1:2 ratio, v / v), CPME / toluene (e.g., in a 1:2 ratio, v / v), NMP / water (e.g., in a 1:4 ratio, v / v), THF / water (e.g., in a 1:4 ratio, v / v), ACN / water (e.g., in a 1:2 ratio, v / v), IPAc / DCM (e.g., in a 1:1 ratio, v / v), MeOH / toluene (e.g., in a 1:4 ratio, v / v), DCM / MTBE (e.g., in a 1:4 ratio, v / v), and THF / n-heptane (e.g., in a 1:4 ratio, v / v).

[0174] Specifically, for the preparation of the above-mentioned crystal form A, for the slow evaporation method, the solvent can be selected from: EtOH, acetone, IPAc, THF, CPME, anisole, ACN / water (e.g., in a 9:1 ratio, v / v), MeOH / DCM (e.g., in a 1:1 ratio, v / v), acetone / EtOAc (e.g., in a 2:1 ratio, v / v), and THF / water (e.g., in a 4:1 ratio, v / v).

[0175] Specifically, for the preparation of the above-mentioned crystal form A, for the slow cooling method, the solvent can be selected from: CPME, toluene, ACN / toluene (e.g., in a 1:2 ratio, v / v), acetone / n-heptane (e.g., in a 1:1 ratio, v / v), THF / toluene (e.g., in a 1:2 ratio, v / v), MeOH / water (e.g., in a 1:1 ratio, v / v), CHCl3 / MTBE (e.g., in a 1:1 ratio, v / v).

[0176] Specifically, for the preparation of the above crystal form A, for the gas-liquid diffusion method, the good solvent can be selected from: EtOH, THF, DMSO, and the antisolvent can be selected from: n-heptane, MTBE, toluene, cyclohexane, and water.

[0177] In one embodiment of the present invention, in the gas-liquid diffusion method, the good solvent is EtOH, and the antisolvent can be selected from: n-heptane, MTBE, and toluene.

[0178] In another embodiment of the present invention, in the gas-liquid diffusion method, the good solvent is THF, and the antisolvent can be selected from: n-heptane, cyclohexane, and MTBE.

[0179] In another embodiment of the present invention, in the gas-liquid diffusion method, the good solvent is DMSO, and the antisolvent can be selected from toluene, MTBE, and water.

[0180] Specifically, for the preparation of the above-mentioned crystal form A, for the polymer-induced method, the solvent can be selected from: MEK, ACN / toluene (e.g., in a 4:1 ratio, v / v), THF / water (e.g., in a 9:1 ratio, v / v), EtOAc, acetone / 2-MeTHF (e.g., in a 1:1 ratio, v / v), MeOH / DCM (e.g., in a 1:1 ratio, v / v), and the polymer can be selected from: polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose, methylcellulose, polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate, and hydroxyethyl cellulose, or one or more combinations thereof.

[0181] In one embodiment of the invention, in the polymer-induced method, the polymer is a mixture of polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose and methylcellulose (e.g., in a mass ratio of 1:1:1:1:1), and the solvent may be selected from: MEK, ACN / toluene (e.g., in a ratio of 4:1, v / v), THF / water (e.g., in a ratio of 9:1, v / v).

[0182] In another embodiment of the invention, in the polymer-induced method, the polymer is a mixture of polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate and hydroxyethyl cellulose (e.g., in a mass ratio of 1:1:1:1:1), and the solvent may be selected from: EtOAc, acetone / 2-MeTHF (e.g., in a 1:1 ratio, v / v).

[0183] Specifically, the preparation method of crystal form B of the above-mentioned 4-(3-(((2-amino-5-(1-(1-trideuterated methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yne-2-ol is the antisolvent addition method.

[0184] In one embodiment of the present invention, for the preparation of the above-mentioned crystal form B, the good solvent is EtOAc and the antisolvent is toluene.

[0185] The present invention also provides a pharmaceutical composition comprising a compound of the above general formula I or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or crystal form thereof, and a pharmaceutically acceptable excipient.

[0186] Specifically, the excipients are selected from one or more of the following: carriers, diluents, binders, lubricants, wetting agents, etc. Specifically, the pharmaceutical compositions comprise a therapeutically effective amount of a compound of formula I. In some embodiments, these pharmaceutical compositions can be used to treat HPK1 kinase-mediated diseases or conditions.

[0187] Specifically, the above-mentioned pharmaceutical compositions may be tablets (e.g., sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, oral tablets, etc.), pills, powders, granules, capsules (e.g., soft capsules, microcapsules), lozenges, syrups, emulsions, suspensions, controlled-release formulations (e.g., instantaneous-release formulations, sustained-release formulations, sustained-release microcapsules), aerosols, films (e.g., orally disintegrating films, oral mucosa-adhesive films), injections (e.g., subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection), intravenous infusions, transdermal absorption formulations, ointments, lotions, adhesive formulations, suppositories (e.g., rectal suppositories, vaginal suppositories), pills, nasal preparations, pulmonary preparations (inhalers), eye drops, etc.

[0188] Specifically, the various dosage forms of the above-mentioned pharmaceutical compositions can be prepared according to conventional pharmaceutical manufacturing methods. For example, the active ingredient is mixed with one or more excipients and then formulated into the desired dosage form.

[0189] Specifically, the above-mentioned pharmaceutical composition may contain an active ingredient in a weight ratio of 0.1-99.5% (e.g., 0.1%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5%).

[0190] The present invention also provides the use of compounds of the above general formula I and their pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the crystal forms described above, or the pharmaceutical compositions described above, in the preparation of medicaments for the prevention and / or treatment of tumors.

[0191] The present invention also provides the use of compounds of the above general formula I and their pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, and the above crystal forms in combination with PD-1, PD-L1, CTLA-4, TIM-3, TGF-β and their receptors, LAG3 antagonists or TLR4, TLR7, TLR8, TLR9, STING agonists in tumor immunotherapy.

[0192] The present invention also provides the use of compounds of the above general formula I, as well as pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, and the above crystal forms in combination with CAR-T immunotherapy in tumor immunotherapy.

[0193] Specifically, the aforementioned CAR-T immunotherapy refers to chimeric antigen receptor T-cell immunotherapy, which is based on the principle of using the patient's own immune cells to eliminate cancer cells and is a type of cell therapy.

[0194] Specifically, the aforementioned tumors are malignant tumors, including but not limited to: lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuroendocrine tumors, carcinoid tumors, gastrinoma, islet cell carcinoma, mesothelioma, schwannoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignant tumors, squamous cell carcinoma, epithelial squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, adenocarcinoma lung cancer, squamous cell carcinoma of the lung, peritoneal carcinoma, hepatocellular carcinoma, gastric cancer, intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, liver cancer, breast cancer, metastatic breast cancer, colon cancer, rectal cancer, colorectal cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, Merkel cell carcinoma, esophageal cancer, biliary tract tumors, head and neck cancers, and hematologic malignancies.

[0195] The present invention also provides the use of compounds of the above general formula I and their pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the crystal forms, or the pharmaceutical compositions thereof, in the preparation of medicaments for the prevention and / or treatment of diseases caused by or related to pathogen infection.

[0196] Specifically, the aforementioned pathogens can be microorganisms, parasites (protozoa, worms, etc.) or other vectors. Specifically, the aforementioned microorganisms can be selected from one or more of the following: viruses, chlamydia, rickettsia, mycoplasma, bacteria, spirochetes, fungi, etc.

[0197] In one embodiment of the present invention, the aforementioned pathogen is a virus, for example, but not limited to, adenoviridae (e.g., adenovirus), herpesviruses (e.g., HSV1 (oral herpes), HSV2 (genital herpes), VZV (varicella), EBV (Ebola virus), CMV (cytomegalovirus)), poxviruses (e.g., smallpox virus, vaccinia virus), polyvacuoviruses (e.g., human papillomavirus (HPV)), parvoviruses (e.g., B19 virus), hepatotropic DNA viruses (e.g., hepatitis B virus (HBV)), polyomaviruses (e.g., polyomavirus), reoviruses (e.g., reovirus, rotavirus), piconenoviruses (e.g., enterovirus, foot-and-mouth disease virus), caliciviruses (e.g., norovirus, hepatitis E virus), clovenviridae (e.g., rubella virus), arenaviruses (e.g., lymphocytic choriomeningitis virus), retroviridae (HIV), flaviviridae (e.g., dengue virus, etc.). Zika virus, Japanese encephalitis virus, Chikungunya virus, yellow fever virus, hepatitis C virus (HCV), West Nile virus, etc.), Orthomyxoviridae (such as influenza viruses (such as influenza A virus, influenza B virus, influenza C virus, etc.)), Paramyxoviridae (such as human parainfluenza virus type 1 (HPV), HPV type 2, HPV type 3, HPV type 4, Sendai virus, mumps virus, measles virus, respiratory syncytial virus, Newcastle disease virus, etc.), Bunyaviridae (such as California encephalitis virus, Hantavirus), Rhabdoviridae (such as rabies virus), Filoviridae (such as Ebola virus, Marburg virus), Coronaviridae (such as HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, SARS-CoV-2, etc.), Astroviridae (such as astrovirus), Bornaviridae (such as Borna virus).

[0198] Specifically, the viruses used in the above applications include HBV, HIV, HCV, HPV, Ebola virus, Marburg virus, influenza virus, parainfluenza virus, dengue virus, SARS-CoV, SARS-CoV-2, etc.

[0199] Specifically, the diseases caused by or related to pathogen infection include, but are not limited to, influenza, SARS, COVID-19, viral hepatitis (such as hepatitis B, hepatitis C, etc.), AIDS, dengue fever, Ebola virus disease, Marburg virus disease, etc.

[0200] The present invention also provides a method for preventing and / or treating tumors, comprising the step of administering to a subject in need an effective amount of a compound of general formula I of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the crystal forms thereof, or the pharmaceutical compositions thereof of the present invention.

[0201] Specifically, a tumor has the corresponding definition described above in this invention.

[0202] The present invention also provides a method for preventing and / or treating diseases caused by or related to pathogen infection, comprising administering to a subject in need an effective amount of a compound of general formula I of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the crystal forms thereof, or the pharmaceutical compositions thereof of the present invention.

[0203] Specifically, pathogens and diseases have the corresponding definitions given above in this invention. Attached Figure Description

[0204] Figure 1 The image shown is the XRPD pattern of the free basal crystal form A of compound A2.

[0205] Figure 2 The image shows the TGA / DSC spectrum of the free alkali crystal form A of compound A2.

[0206] Figure 3 The image shows the free basal crystal form A of compound A2. 1 H NMR spectrum.

[0207] Figure 4 The figure shows the HPLC chromatogram of compound A2 in its free basal form, A.

[0208] Figure 5 The image shows the initial sample of compound A2 in free alkali crystal form A and its XRPD pattern after being placed in a closed container at 60°C for 1 day.

[0209] Figure 6 The image shown is the XRPD pattern of compound A2 in its free basal crystal form B.

[0210] Figure 7 The image shown is the TGA / DSC spectrum of compound A2 in its free basal crystal form B.

[0211] Figure 8 The image shows the free basal crystal form B of compound A2.1 H NMR spectrum.

[0212] Figure 9 The image shows the XRPD spectra of compound A2 in its free alkali crystal form B before and after being placed at room temperature.

[0213] Figure 10 The image shows the XRPD spectra of compound A2 in its free alkali crystal form B before and after nitrogen purging.

[0214] Figure 11 The image shown is the XRPD spectrum of compound A2 hydrochloride.

[0215] Figure 12 The TGA / DSC spectrum of compound A2 hydrochloride is shown.

[0216] Figure 13 The image shows an overlay of the XRPD spectra of the free alkali crystal form A of compound A2 and the hydrochloride salt of compound A2. Detailed Implementation

[0217] Unless otherwise defined, all scientific and technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art.

[0218] In this invention, the term "C" 0-10 "alkyl", C0 alkyl refers to H, therefore, C 0-10 Alkyl groups include H, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, and C 10 alkyl.

[0219] In this invention, the term "C" 1-10 "Straight-chain / branched alkyl", including methyl, ethyl, C3 straight-chain / branched alkyl, C4 straight-chain / branched alkyl, C5 straight-chain / branched alkyl, C6 straight-chain / branched alkyl, C7 straight-chain / branched alkyl, C8 straight-chain / branched alkyl, C9 straight-chain / branched alkyl, C 10 Straight-chain / branched alkyl groups.

[0220] In this invention, the term "C" 3-10 "Branched alkyl groups" include isopropyl, isobutyl, tert-butyl, and isopentyl.

[0221] In this invention, the term "C" 3-10 "Cycloalkyl" includes C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, C8 cycloalkyl, C9 cycloalkyl, C 10 Cycloalkyl.

[0222] In this invention, the term "C" 3-8"Cycloalkyl" includes C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, and C8 cycloalkyl.

[0223] In this invention, the term "C" 4-8 "Cycloalkyl" includes C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, and C8 cycloalkyl.

[0224] In this invention, the term "C" 4-6 "Cycloalkyl" includes C4 cycloalkyl, C5 cycloalkyl, and C6 cycloalkyl.

[0225] The term "halogen" in this invention includes fluorine, chlorine, bromine, and iodine.

[0226] In this invention, the term "heterocyclic alkyl" refers to a non-aromatic saturated monocyclic or polycyclic cyclic system containing 3-10 ring atoms, preferably 5-10 ring atoms, wherein one or more ring atoms are not carbon atoms, but, for example, nitrogen, oxygen, or sulfur atoms. Preferred heterocyclic alkyl systems contain 5-6 ring atoms. The prefixes nitro, oxo, or thio, preceding the heterocyclic alkyl group, respectively, indicate that at least one nitrogen, oxygen, or sulfur atom is a ring atom.

[0227] In this invention, the term "heterocyclic aromatic group" refers to an aromatic monocyclic or polycyclic ring system containing 5-14 ring atoms, preferably 5-10 ring atoms, wherein one or more ring atoms are not carbon atoms, but, for example, nitrogen, oxygen, or sulfur atoms. Preferred heterocyclic aromatic groups contain 5-6 ring atoms. Representative heterocyclic aromatic groups include pyrazinyl, furanyl, thiopheneyl, pyridinyl, pyrimidinyl, isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl, pyrroleyl, pyrazolyl, triazolyl, 1,2,4-thiadiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, 2,3-diazanaphthyl, imidazo[1,2-a]pyridine, imidazo[2,1-b]thiazolyl, benzofurazanyl, indoleyl, azaindoleyl, benzimidazolyl, benzothiopheneyl, quinolinyl, imidazolyl, thiophenepyridinyl, quinazolinyl, thiophenepyrimidinyl, pyrrolopyridinyl, imidazopyridinyl, isoquinolinyl, benzoazaindoleyl, 1,2,4-triazinyl, benzothiazolyl, etc.

[0228] In this invention, "D" refers to deuterium; "replaced by deuterium" means replacing one or more hydrogen atoms with a corresponding number of deuterium atoms.

[0229] It should be recognized that, depending on the source of the chemical materials used in the synthesis, there are variations in the natural isotopic abundance in the synthesized compounds. Therefore, the compounds of the present invention will inherently contain small amounts of deuterated isotopes. Despite this variation, the concentrations of these naturally abundant stable hydrogen and carbon isotopes are low and insignificant compared to the degree of stable isotopic substitution in the compounds of the present invention. See, for example, Wada, E et al., Seikagaku, 1994, 66:15; Gannes, LZ et al., Comp Biochem Physiol Mol Integr Physiol, 1998, 119:725.

[0230] In the compounds of this invention, any atom not specifically designated as deuterium is present at its natural isotopic abundance. Unless otherwise stated, when a position is specifically designated as "H" or "hydrogen", that position should be understood as having hydrogen according to its natural abundance isotopic composition. Similarly, unless otherwise stated, when a position is specifically designated as "D" or "deuterium", that position should be understood as having deuterium at an abundance at least 3000 times greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 45% deuterium doping).

[0231] The term “isotope enrichment coefficient” used in this article refers to the ratio between the isotopic abundance of a particular isotope and its natural abundance.

[0232] In other embodiments, the compounds of the present invention have an isotopic enrichment factor for each specified deuterium atom of at least 3500 (52.5% deuterium doping at each specified deuterium atom), at least 4000 (60% deuterium doping), at least 4500 (67.5% deuterium doping), at least 5000 (75% deuterium doping), at least 5500 (82.5% deuterium doping), at least 6000 (90% deuterium doping), at least 6333.3 (95% deuterium doping), at least 6466.7 (97% deuterium doping), at least 6600 (99% deuterium doping), or at least 6633.3 (99.5% deuterium doping).

[0233] The term "isotope" refers to a substance whose chemical structure differs from that of a specific compound of the present invention only in terms of its isotopic composition.

[0234] The term "compound," when referring to the compounds of this invention, means a collection of molecules having the same chemical structure except that there may be isotopic variations among the constituent atoms of the molecules. Therefore, those skilled in the art will appreciate that a compound represented by a specific chemical structure containing a specified deuterium atom also contains a smaller amount of isotopes having hydrogen atoms at one or more specified deuterium positions in that structure. The relative amount of such isotopes in the compounds of this invention will depend on a variety of factors, including the isotopic purity of the deuterating agent used to make the compound and the efficiency of deuterium incorporation in the various synthetic steps used to prepare the compound. However, as described above, the overall relative amount of such isotopes will be less than 49.9% of the compound. In other embodiments, the overall relative amount of such isotopes will be less than 47.5%, less than 40%, less than 32.5%, less than 25%, less than 17.5%, less than 10%, less than 5%, less than 3%, less than 1%, or less than 0.5% of the compound.

[0235] The explanations of some of the abbreviations used in this invention are as follows:

[0236] XRPD: X-ray powder diffraction

[0237] DSC: Differential Scan Calorimetry

[0238] TGA: Thermogravimetric Analysis

[0239] 1 1H NMR: Liquid NMR spectrum

[0240] In this invention, the term "crystal form" is confirmed by characterization using X-ray powder diffraction (XRD). Those skilled in the art will understand that the physicochemical properties discussed herein can be characterized, with experimental errors depending on instrument conditions, sample preparation, and sample purity, etc. In particular, it is known to those skilled in the art that XRD patterns typically change with instrument conditions. It is particularly important to note that the relative intensities of XRD patterns can also vary with experimental conditions, so the order of peak intensities cannot be considered the sole or decisive factor. In fact, the relative intensities of diffraction peaks in XRPD patterns are related to the preferred orientation of the crystal, and the peak intensities shown herein are illustrative rather than for absolute comparison. Furthermore, experimental errors in peak angles are typically 5% or less, and these angular errors should also be taken into account, generally allowing for ±0.2° errors. Additionally, the overall shift in peak angles due to experimental factors such as sample thickness is generally permissible. Therefore, those skilled in the art will understand that the X-ray powder diffraction pattern of a crystal form in this invention need not be completely identical to the X-ray powder diffraction pattern in the embodiments referred to herein. The phrase "same XRPD pattern" as used herein does not mean absolutely identical; the positions of the same peaks may differ by ±0.2°, and the peak intensities are allowed to have some variability. Any crystal form with a pattern having the same or similar characteristic peaks as those in these spectra falls within the scope of this invention. Those skilled in the art can compare the spectra listed in this invention with a spectra of an unknown crystal form to verify whether the two sets of spectra reflect the same or different crystal forms.

[0241] In some embodiments, the crystal form A of the present invention is pure and singular, substantially free of any other crystal form. In the present invention, "substantially free" when used to refer to a new crystal form means that the crystal form contains less than 20% (by weight) of other crystal forms, particularly less than 10% (by weight) of other crystal forms, more specifically less than 5% (by weight) of other crystal forms, and even more specifically less than 1% (by weight) of other crystal forms.

[0242] It should be noted that the numerical values ​​and ranges mentioned in this invention should not be narrowly interpreted as the numerical values ​​or ranges themselves. Those skilled in the art should understand that they may fluctuate around specific numerical values ​​depending on the specific technical environment, without departing from the spirit and principles of this invention. In this invention, such fluctuation ranges that are foreseeable by those skilled in the art are often expressed using the term "about". When the term "about" is used before a numerical value in this invention and refers to the stated numerical value, it means any value within ±10%, preferably ±5%, more preferably ±2%, and more preferably ±1%. For example, "about 10" should be interpreted as 9-11, preferably 9.5-10.5, more preferably 9.8-10.2, and even more preferably 9.9-10.1.

[0243] In this invention, the term "room temperature" refers to the temperature of an article being close to or the same as the temperature of a space (e.g., the location of a fume hood in which the article is located). Typically, room temperature is about 20°C to about 30°C, or about 22°C to 27°C, or about 25°C.

[0244] Antisolvent addition (also known as antisolvent crystallization, precipitation crystallization, salting-out, or forced crystallization) is a method that typically involves adding one or more antisolvents to a solution in which the target product has been dissolved in a good solvent. The product is in a slightly soluble state in the solution, causing the solution to become supersaturated and crystals to precipitate. Anti-antisolvent addition typically involves adding one or more antisolvents to a solution in which the target product has been dissolved in a good solvent. The product is in a slightly soluble state in the solution, causing the solution to become supersaturated and crystals to precipitate.

[0245] Antisolvents are less effective at dissolving target products than good solvents, for example, by more than 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80%. Therefore, the term "antisolvent" in a system is relative. Good solvents and antisolvents can be polar or nonpolar solvents, such as those selected from one or more of the following: dimethylformamide (DMF), dimethyl sulfoxide (DMSO), water, alcohols, ethers, ketones, esters, alkanes, aromatics, and nitriles. Among them, alcohol solvents include, but are not limited to, methanol, ethanol, propanol, isopropanol, or 1,3-propanediol, 1,2-propanediol, or trichlorotert-butanol or combinations thereof; ether solvents include, but are not limited to, tetrahydrofuran, methyl tert-butyl ether, or 1,4-dioxane or combinations thereof; ketone solvents include, but are not limited to, acetone, methyl ethyl ketone, or 4-methyl-2-pentanone or combinations thereof; ester solvents include, but are not limited to, ethyl acetate, isopropyl acetate, n-butyl acetate, or tert-butyl acetate or combinations thereof; alkane solvents include, but are not limited to, dichloromethane, chloroform, n-hexane, cyclohexane, or pentane or n-heptane or combinations thereof; aromatic solvents include, but are not limited to, benzene, toluene or combinations thereof; and nitrile solvents include, but are not limited to, acetonitrile and malononitrile.

[0246] Antisolvent addition and anti-antisolvent addition can be performed through intermittent, semi-intermittent, or continuous crystallization operations. Antisolvent addition to the solution (antisolvent crystallization) or product solution addition to the antisolvent (anti-antisolvent crystallization) can be done dropwise at a constant rate or by starting slowly and then gradually increasing the rate.

[0247] All publications, patents, and published patent specifications cited in this article are incorporated herein in their entirety through citation.

[0248] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0249] Example 1: Synthesis of compound A2

[0250]

[0251] The experimental steps are as follows:

[0252] Step 1:

[0253]

[0254] In a 500 mL single-necked flask, 1 (11.3 g, 23.7 mmol), pinacol diborate (9.06 g, 35.6 mmol), bis(diphenylphosphine)ferrocene palladium dichloride (1.74 g, 2.37 mmol), potassium acetate (6.99 g, 71.3 mmol), and dimethyl sulfoxide (150 mL) were added. The mixture was reacted at 95 °C for 16 hours under nitrogen protection. The reaction was quenched with water (300 mL), extracted with ethyl acetate (150 mL × 3), washed with saturated brine (150 mL × 2), and evaporated to dryness to obtain a black solid as the target product (13.0 g, crude product). LC-MS: 463 [M + Na] +

[0255] Step 2:

[0256]

[0257] In a 500 mL three-necked flask, 15 g (74.6 mmol), triethylamine (22.7 g, 223.8 mmol), and 150 mL of dichloromethane were added. Methylsulfonyl chloride (12.1 g, 111.9 mmol) was added under ice bath conditions, and the reaction was continued under ice bath conditions for 1 hour. The reaction was quenched with water (300 mL), extracted with dichloromethane (100 mL × 3), washed with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and purified by column chromatography (petroleum ether:ethyl acetate = 40:1) to obtain a yellow solid as the target product (20.7 g, crude product). LC-MS: 280 [M + H] +

[0258] Step 3:

[0259]

[0260] In a 500 mL single-necked flask, 7.27 g (49.4 mmol) and 200 mL of DMF were added. NaH (2.96 g, 74.1 mmol) with a 60% concentration was added in portions at 0 °C. The reaction was carried out at room temperature for 1 hour. Then, 20.7 g (74.1 mmol) was added, and the reaction was carried out under nitrogen protection at 70 °C for 16 hours. The reaction was quenched with 500 mL of water, extracted with ethyl acetate (200 mL × 3), and the organic phase was washed with saturated brine (200 mL × 2). The mixture was dried over anhydrous sodium sulfate, evaporated to dryness, and separated by column chromatography (petroleum ether: ethyl acetate = 30:1) to give a white solid as the target product (9.8 g, yield: 61.2%). LC-MS: 330 [M+H]+

[0261] Step 4:

[0262]

[0263] In a 500 mL single-necked flask, 6 (9.8 g, 35.8 mmol) was dissolved in DCM (80 mL), and TFA (16 mL) was added dropwise at 0 °C. The reaction was allowed to proceed for 16 h at room temperature. The solution was concentrated at low temperature, diluted with DCM (200 mL), quenched with ice water, and the pH was adjusted to 10 with ammonia at 0 °C. The solution was extracted with DCM (200 mL × 3), and the organic phase was washed with saturated brine (200 mL × 2), dried over anhydrous sodium sulfate, and evaporated to dryness to give 6.59 g of the target product as a white solid (yield: 96.6%). LC-MS: 230 [M + H] +

[0264] Step 5:

[0265]

[0266] In a 250 mL three-necked flask, 7 (5.53 g, 24.0 mmol) and TEA (10 mL, 72.1 mmol) were dissolved in THF (80 mL), and the reaction was carried out at room temperature for 1 hour. The mixture was then cooled to 0 °C in an ice bath, and CD3I (1.65 mL, 26.4 mmol) was added dropwise. Under nitrogen protection, the reaction was carried out at room temperature for 2 hours. The reaction was quenched with water, extracted with DCM (100 mL × 3), and the organic phase was washed with saturated brine (100 mL × 2). The mixture was dried over anhydrous sodium sulfate and evaporated to dryness to obtain a yellow oily substance, 3.1 g of which was the target product (yield: 52.2%). LC-MS: 248 [M + H] +

[0267] Step 6:

[0268]

[0269] In a 500 mL single-necked flask, 2 (7.13 g, 16.2 mmol), 8 (2.67 g, 10.8 mmol), xphosPdGⅡ (850 mg, 1.08 mmol), xphos (515 mg, 1.08 mmol), potassium phosphate (4.58 g, 21.6 mmol), and DMF / H2O (150 mL / 30 mL) were added. The mixture was reacted under nitrogen protection at 95 °C for 2.5 h. The reaction was quenched with water (300 mL), extracted with ethyl acetate (150 mL × 3), washed with saturated brine (150 mL × 2), dried over anhydrous sodium sulfate, and evaporated to dryness. The product (3.3 g, crude) was purified by Flash chromatography to obtain a black oily substance. LC-MS: 563 [M + H] +

[0270] Step 7:

[0271]

[0272] In a 250 mL three-necked flask, 3.3 g (7.14 mmol), 2.0 g (35.7 mmol), 1.93 g (35.7 mmol), 40 mL of ethanol, and 8 mL of water were added, and the mixture was reacted at 85 °C for 2.5 hours. The mixture was filtered, and the filtrate was concentrated to obtain 3.5 g of a black solid crude product as the target product. LC-MS: 533 [M+H]+

[0273] Step 8:

[0274]

[0275] In a 250 mL three-necked flask, 10 (3.5 g, 6.55 mmol) was dissolved in tetrahydrofuran (40 mL), and HCl / Dioxane (8 mL) was added dropwise at 0 °C. The reaction was carried out at room temperature for 1 hour. The solution was concentrated at low temperature, diluted with DCM (100 mL), quenched with ice water, and the pH was adjusted to 10 with ammonia at 0 °C. The solution was extracted with DCM (100 mL × 3), and the organic phase was washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, and evaporated to dryness. The solution was purified by Prep-HPLC to give 450 mg of the target product as a white solid (yield: 15.3%). LC-MS: 449[M+H]+,1H NMR (400MHz, MeOD) δ8.38(s,1H),8.01(s,1H),7.81(s,1H),7.73(d,J=1.5Hz,1H),7.54(s,1H),7.48(td,J=4.7,1.7Hz,1 H),7.38(s,2H),5.21(s,2H),4.51(t,J=6.9Hz,1H),3.59(d,J=12.6Hz,2H),3.19(s,2H),2.37–2.32(m,4H),1.56(s,6H).

[0276] Example 2: Synthesis of compound B2

[0277]

[0278] The experimental steps are as follows:

[0279] Step 1:

[0280]

[0281] In a 2000 mL single-necked flask, 25.0 g (102 mmol) of methyl chloroacetaldehyde (12.0 g, 154 mmol), 12.0 g (154 mmol) of aqueous chloroacetaldehyde, and 500 mL of acetone were added, and the reaction was carried out at 50 °C for 16 hours. The solution was evaporated to dryness and separated by column chromatography (MeOH in DCM, from 0% to 10%, v / v) to give a yellow oily product (9.20 g, yield: 53.4%). LC-MS: 169 [M+H]+

[0282] Step 2:

[0283]

[0284] In a 1000 mL single-necked flask, 3 (4.00 g, 23.8 mmol), triethylamine (6.01 g, 59.5 mmol), and tetrahydrofuran (200 mL) were added. Deuterated iodomethane (3.62 g, 25.0 mmol) was added at room temperature, and the mixture was stirred for 2 hours at room temperature. The mixture was quenched with water (250 mL), evaporated to dryness, extracted with ethyl acetate (150 mL x 3), dried over anhydrous sodium sulfate, and evaporated to dryness again. Column chromatography (MeOH in DCM, from 0% to 10%, v / v) yielded a yellow solid as the target product (2.70 g, yield: 61.3%). LC-MS: 186 [M+H]+

[0285] Step 3:

[0286]

[0287] Add 4 (2.70 g, 14.6 mmol) and tetrahydrofuran (100 mL) to a 500 mL three-necked flask. Under nitrogen protection, add n-butyllithium (2.4 M in tetrahydrofuran, 7.30 mL, 17.5 mmol) dropwise at -78 °C, maintaining the temperature and stirring for 1 h. Then add tri-n-butylstannous chloride (7.14 g, 21.9 mmol) dropwise, and continue the reaction at -78 °C for 1 h. After the reaction is complete, quench the reaction with saturated ammonium chloride aqueous solution (100 mL), extract with ethyl acetate (120 mL x 3), wash the organic layer with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a yellow oily substance without the target product. This can be used directly for the next step. (7.05 g, crude product). LC-MS: 476 [M + H] +

[0288] Step 4:

[0289]

[0290] Add 6 (59.4 g, 707 mmol) and tetrahydrofuran (1.5 L) to a 3 L single-necked flask, followed by DHP (68.4 g, 813 mmol) and PPTS (3.55 g, 14.1 mmol), and react at room temperature for 16 hours. Wash with saturated sodium bicarbonate (1000 mL * 3), then with saturated brine (500 mL), dry to anhydrous sodium sulfate, and evaporate to dryness to obtain a colorless oily product (120 g, crude product), which can be used directly in the next reaction. LC-MS: 169 [M + H] +

[0291] Step 5:

[0292]

[0293] In a 3L single-necked flask, 7 (35.8 g, 213.15 mmol), 8 (47.5 g, 203.00 mmol), bis(triphenylphosphine)palladium dichloride (1.43 g, 2.03 mmol), cuprous iodide (1.93 g, 10.16 mmol), triethylamine (40.01 g, 406.00 mmol), and anhydrous dichloromethane (1 L) were added. The reaction mixture was incubated at room temperature for 16 hours under nitrogen protection. The reaction solution was washed with saturated ammonium chloride (1000 mL x 3) and saturated brine (500 mL), dried over anhydrous sodium sulfate, and evaporated to dryness to obtain a yellow oily substance, which was the target compound (59.4 g, crude product), which could be used directly in the next reaction. LC-MS: 275 [M + H] +

[0294] Step 6:

[0295]

[0296] In a 2000 mL three-necked flask, 9 (50.3 g, 184 mmol), tetrahydrofuran (500 mL), 10 (40.1 g, 184 mmol), and triphenylphosphine (72.3 g, 276 mmol) were added under nitrogen protection. Diethyl azodicarbonate (55.8 g, 276 mmol) was added with stirring at room temperature. The reaction was carried out at room temperature for 16 hours, and the solution was evaporated to dryness. Column chromatography (petroleum ether: ethyl acetate = 10:1) yielded a yellow solid as the target product (70.2 g, yield: 80.6%). LC-MS: 476 [M+H] +

[0297] Step 7:

[0298]

[0299] In a 1000 mL single-necked flask, 11 (5.05 g, 10.7 mmol), 5 (9.15 g, 19.3 mmol), bis(triphenylphosphine)palladium(II) dichloride (376 mg, 0.535 mmol), cuprous iodide (305 mg, 1.61 mmol), and 1,4-dioxane (200 mL) were added. The mixture was stirred at 90 °C for 5 hours under nitrogen protection. The solution was diluted with ethyl acetate (400 mL), washed with saturated ammonium chloride aqueous solution (300 mL x 3), evaporated to dryness, and separated by column chromatography (MeOH in DCM, from 0% to 5%, v / v) to give a yellow solid as the target product (2.50 g, yield: 40.5%). LC-MS: 580 [M+H] +

[0300] Step 8:

[0301]

[0302] In a 500 mL single-necked flask, 2.50 g (4.32 mmol) of 12, 1.21 g (21.6 mmol) of reduced iron powder, 1.14 g (21.6 mmol) of ammonium chloride, 100 mL of ethanol, and 20 mL of water were added and reacted at 80 °C for 2 hours. The mixture was then diluted with 100 mL of dichloromethane, filtered, evaporated to dryness, and separated by column chromatography (MeOH in DCM, from 0% to 12%, v / v) to obtain a yellow solid as the target product (1.40 mg, 59.1%). LC-MS: 550 [M+H]+

[0303] Step 9:

[0304]

[0305] Add 13 (1.02 g, 1.86 mmol) and tetrahydrofuran (25 mL) to a 250 mL single-necked flask, stir under ice bath, then add dropwise a 1,4-dioxane solution (5 mL) of 4 M / L hydrochloric acid gas. React at room temperature for 20 minutes, evaporate to dryness, dissolve in dichloromethane, adjust pH to 9 with ammonia, evaporate to dryness with dichloromethane (30 mL x 3), and prepare the target product (350 mg, 40.5%) by high-performance liquid chromatography. LC-MS: 466 [M+H]+ 1 H NMR (400MHz, DMSO) δ7.88(s,1H),7.77(d,J=1.8Hz,1H),7.54(d,J=5.4Hz,2H),7.42–7.37(m,1H),7.34(dd,J=5.4,1.6Hz,2H),6. 10(s,2H),5.50(s,1H),5.22(s,2H),2.96–2.78(m,3H),2.02(dd,J=16.9,6.7Hz,4H),1.72(qd,J=12.5,3.6Hz,2H),1.47(s,6H).

[0306] Example 3: Synthesis of Compound C

[0307]

[0308] The experimental steps are as follows:

[0309] Step 1:

[0310]

[0311] Add 1 (7.86 g, 30.0 mmol) and tetrahydrofuran (500 mL) to a 2000 mL single-necked flask. Add lithium aluminum deuterated hydride (3.15 g, 75.0 mmol) in portions at 0 °C, and react at 0 °C for 1 hour. Quench with acetic acid (50 mL), extract with ethyl acetate (500 mL × 3), wash with saturated brine (500 mL × 3), and evaporate to dryness to obtain a yellow solid as the target product (3.50 g, crude product). LC-MS: 259 [M + Na] +

[0312] Step 2:

[0313]

[0314] In a 250 mL three-necked round-bottom flask, 3 (3.50 g, 14.8 mmol), bis(triphenylphosphine)palladium dichloride (519 mg, 0.740 mmol), cuprous iodide (281 mg, 1.48 mmol), triethylamine (4.48 g, 44.4 mmol), and dichloromethane (60 mL) were added. Under nitrogen protection at room temperature, 3 (2.50 g, 14.8 mmol) was added, and the reaction was carried out for 16 hours at room temperature. The mixture was diluted with dichloromethane (400 mL), washed with saturated ammonium chloride aqueous solution (300 mL x 3), evaporated to dryness, and subjected to column chromatography (petroleum ether: ethyl acetate = 4:1) to give a yellow solid as the target product (1.03 g, yield: 25.2%). LC-MS: 299 [M + Na] +

[0315] Step 3:

[0316]

[0317] Add 4 (1.03 g, 3.73 mmol), tetrahydrofuran (30 mL), 5 (854 mg, 3.92 mmol), and triphenylphosphine (1.47 g, 5.60 mmol) to a 250 mL three-necked flask. Add diethyl propyl azodicarbonate (1.13 g, 5.60 mmol) dropwise under nitrogen protection at 0 °C. React at room temperature under nitrogen protection for 16 hours. Dry to dryness, and obtain the target product (1.30 g, yield: 73.2%) by column chromatography (petroleum ether: ethyl acetate = 10:1). LC-MS: 499 [M+Na]+

[0318] Step 4:

[0319]

[0320] In a 100 mL single-necked flask, 6 (300 mg, 0.630 mmol), bis-pinacol borate (240 mg, 0.945 mmol), Pd(dppf)Cl2 (23.5 mg, 0.0315 mmol), potassium acetate (154 mg, 1.58 mmol), and dimethyl sulfoxide (10 mL) were added. The mixture was reacted at 90 °C for 16 hours under a nitrogen atmosphere. The reaction was quenched with water (80 mL), extracted with ethyl acetate (50 mL x 3), washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, and evaporated to dryness to obtain a brown oily product (400 mg, crude). LC-MS: 443 [M + H] +

[0321] Step 5:

[0322]

[0323] In a 100 mL single-necked flask, 7 (400 mg, 0.905 mmol), 8 (200 mg, 0.905 mmol), XPhosPdG2 (35.6 mg, 0.0453 mmol), XPhos (43.2 mg, 0.0905 mmol), potassium phosphate (384 mg, 1.81 mmol), DMF (10 mL), and water (2 mL) were added. The reaction was carried out under nitrogen protection at 90 °C for 2 hours. The mixture was quenched with water (80 mL), extracted with ethyl acetate (50 mL x 3), washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, and evaporated to dryness. The product was then subjected to column chromatography (dichloromethane:methanol = 18:1) to obtain a yellow solid as the target product (250 mg, yield: 49.2%). LC-MS: 562 [M+H] +

[0324] Step 6:

[0325]

[0326] In a 100 mL single-necked flask, 250 mg (0.446 mmol) of ferric chloride, 125 mg (2.23 mmol) of reduced iron powder, 118 mg (2.23 mmol) of ammonium chloride, 10 mL of ethanol, and 2 mL of water were added and reacted at 80 °C for 2 hours. The mixture was then diluted with 50 mL of dichloromethane, filtered, evaporated to dryness, and separated by column chromatography (dichloromethane:methanol = 10:1) to obtain a yellow solid as the target product (220 mg, 93.0%). LC-MS: 532 [M+H]+

[0327] Step 7:

[0328]

[0329] In a 250 mL single-necked flask, 10 mL of 10 (220 mg, 0.414 mmol) and tetrahydrofuran were added and stirred in an ice bath. Then, a 1,4-dioxane solution (3 mL) of 4 M / L hydrochloric acid was added dropwise. The mixture was reacted at room temperature for 20 minutes, evaporated to dryness, dissolved in dichloromethane, and the pH was adjusted to 9 with ammonia. The solution was then evaporated to dryness with dichloromethane (30 mL x 3). The target product (85.0 mg, 45.9%) was obtained by high-performance liquid chromatography (HPLC). LC-MS: 448 [M+H]+ 1 H NMR(400MHz,DMSO)δ8.10(s,1H),7.79(d,J=1.8Hz,1H),7.75(s,1H),7.58–7.49(m,2H),7.44–7.30(m,3H),5.66 (s,2H),5.49(s,1H),4.14–4.00(m,1H),2.86(d,J=11.5Hz,2H),2.21(s,3H),1.99(d,J=3.1Hz,6H),1.46(s,6H).

[0330] Example 4: Synthesis of Compound D

[0331]

[0332] The experimental steps are as follows:

[0333] Step 1:

[0334]

[0335] Add 1 (3.0 g, 30.61 mmol) and tetrahydrofuran (30 mL) to a 100 mL three-necked flask, under nitrogen protection, and add n-butyllithium (2.4 M in) dropwise at -78 °C. tetrahydrofuran, 12.75 mL 30.61 mmol), keep the temperature constant and continue stirring for 1 hour, then add 2 (2.0 g, 30.61 mmol) dropwise. 78 Continue the reaction at ℃ for 1 hour, then raise the temperature to room temperature and react for another 2 hours. After the reaction is complete, use a saturated ammonium chloride aqueous solution ( 30mL Quenching reaction, ethyl acetate ( 100mL x 3 Extraction was performed, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting yellow oily substance contained no target product and could be used directly in the next step. (6.1 g, crude product). LC-MS: 163 [M+H] +

[0336] Step 2:

[0337]

[0338] Add 6.1 g (37.65 mmol) of methyl methacrylate (MTBF) and 50 mL of tetrahydrofuran to a 250 mL single-necked flask, followed by DHP (4.76 g, 56.63 mmol) and PPTS (158.8 mg, 0.63 mmol), respectively. React at room temperature for 16 hours. Wash with 100 mL x 3 saturated sodium bicarbonate solution, then wash with 100 mL of saturated brine, dry to anhydrous sodium sulfate, and evaporate to dryness to obtain a colorless oily product (8.5 g, crude product), which can be used directly in the next reaction. LC-MS: 247 [M+H] +

[0339] Step 3:

[0340]

[0341] Add 4 (8.5 g, 34.55 mmol), methanol, dichloromethane (10 mL / 10 mL), and potassium carbonate (7.15 g, 51.83 mmol) to a 250 mL single-necked flask, and react at room temperature for 3 hours. Filter, concentrate the filtrate, and evaporate to dryness to obtain a colorless oily substance as the target product (6.3 g, crude product), which can be used directly in the next reaction. LC-MS: 175 [M+H] +

[0342] Step 4:

[0343]

[0344] In a 250 mL single-necked flask, 5 (6.3 g, 36.21 mmol), 6 (8.05 g, 34.40 mmol), bis(triphenylphosphine) palladium dichloride (238 mg, 0.34 mmol), cuprous iodide (322 mg, 1.69 mmol), triethylamine (6.95 g, 68.8 mmol), and anhydrous dichloromethane (80 mL) were added. The reaction mixture was reacted at room temperature for 16 hours under nitrogen protection. The reaction solution was washed with saturated ammonium chloride (200 mL x 3) and saturated brine (100 mL), dried over anhydrous sodium sulfate, and evaporated to dryness. The solution was then subjected to column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain a yellow oily substance as the target compound (3.2 g). LC-MS: 281 [M + H] +

[0345] Step 5:

[0346]

[0347] In a 100 mL three-necked flask, 7 (1.5 g, 5.36 mmol), 8 (1.752 g, 8.04 mmol), triphenylphosphine (2.106 g, 8.04 mmol), and anhydrous tetrahydrofuran (20 mL) were added. Under nitrogen protection, diisopropyl azodicarbonate (1.624 g, 8.04 mmol) was added with stirring at room temperature. The reaction was carried out at room temperature for 16 hours, and the solution was evaporated to dryness. Column chromatography (petroleum ether: ethyl acetate = 4:1) yielded a yellow solid as the target product (450 mg, yield: 17.50%). LC-MS: 481 [M+H] +

[0348] Step 6:

[0349]

[0350] In a 100 mL single-necked flask, 9 (450 mg, 0.94 mmol), pinacol borate (359 mg, 1.41 mmol), bis(diphenylphosphino)ferrocene palladium dichloride (35 mg, 0.047 mmol), potassium acetate (276 mg, 2.82 mmol), and dimethyl sulfoxide (10 mL) were added. The mixture was reacted under nitrogen protection at 90 °C for 16 hours. The reaction was quenched with water (30 mL), extracted with ethyl acetate (50 mL × 3), washed with saturated brine (100 mL × 2), and evaporated to dryness to obtain a black solid as the target product (500 mg, crude product). LC-MS: 447 [M + H] +

[0351] Step 7:

[0352]

[0353] Add 10 (363 mg, 0.81 mmol), 11 (178 g, 0.73 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (59 mg, 0.081 mmol), 2-dicyclohexylphospho-2,4,6-triisopropylbiphenyl (39 mg, 0.082 mmol), potassium phosphate (343 mg, 1.62 mmol), DMF (10 mL), and H2O (2 mL) to a 100 mL single-necked flask, under nitrogen protection, and stir at 95 °C for 1.5 h. Quenching with water (30 mL), extraction with ethyl acetate (100 mL × 3), combining the organic phases and washing with saturated brine (100 mL × 3), drying to anhydrous sodium sulfate, evaporating to dryness, and separating by TCL (dichloromethane:methanol = 15:1) to obtain a brown liquid as the target product (266 mg, yield: 57.9%). LC-MS: 566 [M + H] +

[0354] Step 8:

[0355]

[0356] In a 100 mL single-necked flask, 12 (266 mg, 0.47 mmol), iron powder (201 mg, 3.77 mmol), ammonium chloride (203 mg, 3.77 mmol), ethanol (10 mL), and water (2 mL) were added, and the mixture was reacted at 80 °C for 1 h. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated. Separation was performed by TLC (methanol:dichloromethane = 1:10, v / v), yielding a yellow solid as the target product (95 mg, 37.6%). LC-MS: 536 [M+H] +

[0357] Step 9:

[0358]

[0359] 13 (95 mg, 0.17 mmol) and tetrahydrofuran (5 mL) were added to a 50 mL single-necked flask and stirred in an ice bath. Then, 0.01 mL (0.34 mmol) of 4 M / L 1,4-dioxane hydrochloride solution was added dropwise. The mixture was reacted at room temperature for 1 h, and then evaporated to dryness at 0 °C. The resulting black solid (15 mg, 18.7%) was obtained by high-performance liquid chromatography (HPLC). LC-MS: 452 [M+H] +1 H NMR(400MHz, DMSO)δ8.20(s,1H),8.10(s,1H),7.80(d,J=1.7Hz,1H),7.76(s,1H),7.52(d,J=1.4Hz,2H),7.40(s,1H),7.35–7.32(m,2H),5 .67(s,2H),5.18(s,2H),4.09(dd,J=10.4,5.1Hz,1H),2.89(d,J=11.1Hz,2H),2.24(s,3H),2.10(d,J=11.3Hz,2H),2.00(d,J=3.3Hz,4H).

[0360] Example 5: Free base crystal form A of compound A2

[0361] Compound A2 prepared in Example 1 was purified by silica gel column chromatography to obtain 1.6 g of crude product (purity approximately 80%). This crude product was then separated by preparative high-performance liquid chromatography (HPLC), concentrated to 100 mL, and adjusted to pH 9 with sodium bicarbonate solution. The product was extracted three times with dichloromethane (100 mL). The organic phase was washed once with saturated sodium chloride solution (80 mL), dried over sodium sulfate, and evaporated to dryness to obtain 570 mg of a pale yellow solid (purity 97%). This solid was slurried with 50 mL of petroleum ether / ethyl acetate (3:1), filtered, and yielded 460 mg of an off-white solid (purity >99%).

[0362] Repeated preparation:

[0363] (1) Weigh 230.9 mg of the above free base sample into a 20 mL glass bottle, add 1 mL of IPAc, and obtain a clear solution;

[0364] (2) After stirring at room temperature (1000 rpm) for about 5 minutes, a large amount of solid precipitates out. Then, add 1 mL of IPAc.

[0365] (3) After suspending and stirring at room temperature for about 1 day, centrifuge (10000 rpm, 2 min) to obtain a solid;

[0366] (4) After vacuum drying at room temperature for 3 hours, a total of 195.4 mg of sample was collected (recovery rate: 84.6%).

[0367] The XRPD spectra of the above samples are as follows: Figure 1 As shown, its XRPD test parameters and result data are shown in Tables 1 and 2, respectively, indicating that it is a crystal, named free-state crystal form A, and can be repeatedly prepared. Table 2 contains a complete list of peaks or corresponding d-values, or a subset thereof, and is essentially similar to... Figure 1 The XRPD pattern is sufficient to characterize this crystal form.

[0368] The TGA / DSC spectrum of compound A2 free base crystal form A is as follows: Figure 2 As shown in Table 3, the test parameters for TGA and DSC are as follows. Figure 2 In the study, TGA results showed that it lost 1.3% of its weight when heated from room temperature to 170℃; DSC results showed that it had a sharp endothermic peak at 168.8℃ (initial temperature).

[0369] Compound A2 free base crystal form A 1 H NMR spectrum as follows Figure 3 As shown. 1 1H NMR results showed that the molar ratio of the residual solvent IPAc to the free alkali crystal form A was 0.01:1 (corresponding to a TGA weight loss of 0.2%).

[0370] The HPLC chromatogram of compound A2 free base crystal form A is as follows: Figure 4 As shown in the figure, the results are shown in Table 4.

[0371] It is speculated that the free alkali crystal form A of compound A2 is the amorphous form.

[0372] Table 1 XRPD Test Parameters

[0373]

[0374] Table 2. XRPD data of compound A2 free alkali crystal form A.

[0375]

[0376]

[0377] Table 3 Test parameters for TGA and DSC

[0378] parameter TGA DSC method linear heating linear heating Sample tray Aluminum tray, open Aluminum disc, pressure cap Temperature range Ambient temperature -350℃ 25-350℃ heating rate 10℃ / min 10℃ / min Protective gas Nitrogen Nitrogen

[0379] Table 4. HPLC results of free basal form A of compound A2.

[0380] peak RRT area(%) 1 0.95 0.42 2 1.00 99.26 3 1.05 0.22 4 1.06 0.10

[0381] Solid-state stability of free alkali crystal form A

[0382] The XRPD spectra of the above-mentioned free alkali crystal form A starting sample and the sample after being placed in a closed container at 60℃ for 1 day are shown below. Figure 5 As shown in Table 5, the HPLC results are as follows.

[0383] Table 5. HPLC results of compound A2 free basal crystal form A.

[0384]

[0385] Example 6: Compound A2 free base crystal form B

[0386] (1) Weigh 9.9g of free alkali sample (as described in Example 5) into a 20mL glass bottle, add 2mL of EtOAc to dissolve the sample, filter with a 0.45μm PTFE filter membrane to obtain a clear API solution;

[0387] (2) Add the antisolvent Toluene dropwise to the API solution while stirring magnetically (~1000 rpm) until a total of 10 mL of Toluene is added;

[0388] (3) After stirring the clear solution at room temperature for 2 hours, transfer it to 5°C and stir for about 15 hours. The solution is still clear.

[0389] (4) After stirring at -20°C for about 6 hours, the solution remains clear.

[0390] (5) The clear solution was transferred to room temperature and allowed to evaporate for 8 days to obtain a solid.

[0391] The XRPD spectra of the above samples are as follows: Figure 6 As shown in Table 1 above, its XRPD test parameters are shown in Table 6 above, and the result data is shown in Table 6, indicating that it is a crystal, named free-state crystal form B. Table 6 contains a complete list of peaks or corresponding d-values, or a subset thereof, and is essentially similar to... Figure 5The XRPD pattern is sufficient to characterize this crystal form.

[0392] Table 6. XRPD data of compound A2 free alkali crystal form B.

[0393]

[0394] The TGA / DSC spectrum of compound A2 free base crystal form B is as follows: Figure 7 As shown in Table 3 above, the test parameters for TGA and DSC are as follows. The TGA results show that the sample lost 13.2% of its weight when heated from room temperature to 70℃, and 8.5% when heated further to 170℃. The DSC results show that the sample has four endothermic peaks at 59.5℃, 95.6℃, 150.8℃, and 160.9℃ (peak temperature).

[0395] Compound A2 free base crystal form B 1 H NMR spectrum as follows Figure 8 As shown. 1 1H NMR results showed that the molar ratio of residual solvent EtOAc to free base was 0.8:1 (corresponding to a TGA weight loss of 18.1%).

[0396] The XRPD spectra of compound A2 with free base crystal form B before and after being placed at room temperature are as follows: Figure 9 As shown in the figure. The results indicate that after free alkali crystal form B was placed in a closed container at room temperature for 5 days, diffraction peaks of free alkali crystal form A appeared, indicating that free alkali crystal form B tends to transform into free alkali crystal form A after being placed at room temperature.

[0397] The XRPD spectra of compound A2 free base crystal form B before and after nitrogen purging are as follows: Figure 10 As shown in the figure, the results indicate that the free alkali crystal form B (containing one diffraction peak of crystal form A) undergoes a crystal form transformation after being purged with nitrogen at 30 °C for 20 minutes. It is speculated that the free alkali crystal form B loses EtOAc after nitrogen purging, leading to the crystal form transformation.

[0398] Based on the comprehensive characterization results, it is speculated that the free alkali crystal form B is an EtOAc solvate.

[0399] Example 7: Preparation of compound A2 hydrochloride:

[0400] 100 mg (0.223 mmol) of compound A2 was weighed and dissolved in 5 mL of anhydrous dichloromethane. The mixture was stirred at room temperature for 5 minutes, and then 1.11 mL of diethyl ether hydrochloride solution (1N) was added dropwise. After the addition was complete, the mixture was stirred at room temperature for another 30 minutes. TLC showed that the starting material had completely disappeared. Nitrogen gas was purged for 10 minutes, and the mixture was concentrated under reduced pressure at 10 °C to obtain a white solid (purity >99%). This solid was lyophilized for 12 hours to give 108 mg of A2 hydrochloride (white, purity >99%).

[0401] The XRPD spectrum of compound A2 hydrochloride is as follows: Figure 11 As shown, the XRPD test parameters are shown in Table 1 of Example 5, and the XRPD result data is shown in Table 7, indicating that it is a crystal, named hydrochloride crystal form A. Table 7 contains a complete list of peaks or corresponding d-values, or a subset thereof, and is substantially similar to... Figure 11 The XRPD pattern is sufficient to characterize this crystal form.

[0402] The TGA / DSC spectrum of compound A2 hydrochloride is as follows: Figure 12 As shown, the test parameters for TGA and DSC are shown in Table 3 of Example 5. The TGA results show that it loses 8.6% of its weight when heated from room temperature to 150°C; the DSC results show that it has two endothermic peaks at 81.9°C and 156.0°C (peak temperature).

[0403] Table 7. XRPD data for compound A2 hydrochloride.

[0404]

[0405]

[0406] XRPD patterns of compound A2 free base crystal form A and compound A2 hydrochloride are overlaid as follows: Figure 13 As shown, the XRPD results of the two are basically the same, but the crystallinity of the hydrochloride is lower.

[0407] Example 8

[0408] Procedure for identifying microsomal metabolites:

[0409] Weigh out the compounds (test compounds and their numbers are shown below, where compounds A1 and B1 are prepared according to the relevant methods disclosed in CN110396087A) and dissolve them in dimethyl sulfoxide to prepare a 20 mM solution. Dilute the compound stock solution with 50% acetonitrile aqueous solution (v / v) to a concentration of 1.0 mM as the working solution. Dilute liver microsomes (20 mg / mL) with 50 mM dipotassium hydrogen phosphate buffer to a concentration of 1.27 mg / mL as the liver microsome working solution. Weigh out the reduced coenzyme and dissolve it in 3307 μL of phosphate buffer (50 mM) to a concentration of 5.0 mM as the reduced coenzyme working solution. For samples with T = 60 minutes, add 4 μL of the test sample working solution (1.0 mM), then add 316 μL of liver microsome working solution (1.27 mg / mL), and finally add 80 μL of reduced coenzyme working solution to start the reaction. For samples with T = 0 minutes, add 316 μL of liver microsome working solution (1.27 mg / mL), then add 80 μL of reduced coenzyme working solution to start the reaction, without incubation with the test sample. After incubation at 37°C for 60 minutes, add 1200 μL of stop solution to terminate the enzyme reaction. For samples with T = 0 minutes, add 4 μL of the test sample working solution (1.0 mM). Place the sample plate on a vortex mixer and vortex at 600 rpm for 5 minutes. Then centrifuge at 4000 rpm for 10 minutes, transfer the supernatant, mix, and dry under nitrogen at room temperature. The residue was reconstituted with 300 μL of 10% acetonitrile (0.1% FA) solvent, dried under nitrogen, and then centrifuged at 4000 rpm for 15 minutes. The supernatant was transferred to a detection plate for mass spectrometry analysis. Mass spectrometry analysis was performed using LC / Q-Exactive plus.

[0410] 7-Ethoxycoumarin (10 μM) was selected as the positive control compound, and the same operating procedures as those for the compound were followed.

[0411]

[0412] The remaining percentage of the parent nucleus after 60 minutes is shown in the table below:

[0413] Table 8 Results of compound stability studies

[0414] Test species A1(％) A2(％) B1(％) B2(％) mice 1.13 55.0 1.24 42.8 rats 2.44 52.1 11.21 43.9 Beagle 0.17 7.0 0.12 8.9 Crab-eating macaques 0.36 29.6 9.11 23.4 people 43.84 83.9 51.82 70.4

[0415] Example 9 Pharmacokinetic Experiment

[0416] Rat pharmacokinetics experiment

[0417] Six male Sprague Dawley rats were administered single doses, either intravenously (iv, n=3) or orally (po, n=3). For intravenous administration, the compound was prepared into a solution of 0.25 mg / mL or 0.5 mg / mL using 10% DMSO / 30% PEG400 / 60% Water, and administered at a volume of 2 mL / kg. For oral administration, the compound was prepared into a homogeneous suspension of 0.6 mg / mL or 2.0 mg / mL using 0.5% Methylcellulose, and administered at a volume of 5 mL / kg. Specific dosages are shown in the table below.

[0418] Table 9 Dosage

[0419] compound IV (mg / kg) po (mg / kg) A1 1 10 A2 1 10 C 0.5 3 D 0.3 2

[0420] Blood samples were collected intravenously and orally at 0.0833, 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration. The concentration of the compounds in the samples was detected by LC-MS / MS with a limit of quantitation of 1 ng / mL. The pharmacokinetic parameters of the compounds were calculated using the WinNolin non-compartmental model.

[0421] The structures and numbering of the compounds involved in this embodiment are shown below:

[0422]

[0423] result

[0424] In vivo exposure (AUC) of A1, A2, C, and D after intravenous administration. last The effective rates were 116 hr*ng / mL, 247 hr*ng / mL, 74.8 hr*ng / mL, and 28.7 hr*ng / mL, respectively, and the average total clearance (CL) was 129 mL / min / kg, 71 mL / min / kg, 101 mL / min / kg, and 151 mL / min / kg, respectively.

[0425] After oral administration, the in vivo exposure (AUC) of A1, A2, C and D. last The bioavailability rates (BRPs) in rats were 79.8 ng / hr, 572 ng / hr, 42.8 ng / hr, and 9.94 ng / hr, respectively. Compared with the intravenous data, the oral bioavailability rates in rats were 6.84%, 23.2%, 9.52%, and 5.2%, respectively.

[0426] Pharmacokinetics of cynomolgus monkeys

[0427] Three male cynomolgus macaques were administered a single dose intravenously (iv, 1 mg / kg, n=3), followed by oral administration one week later (po, 10 mg / kg, n=3). For intravenous administration, the compound was reconstituted into a 1.0 mg / mL solution in water or 10% DMSO / 30% PEG400 / Water and administered at a volume of 1 mL / kg. For oral administration, the compound was reconstituted into a 2.0 mg / mL solution or suspension in water or 0.5% methylcellulose / 1.7 meq 1N hydrochloric acid and administered at a volume of 5 mL / kg.

[0428] Following administration, intravenous and oral blood samples were collected at 0.0833, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours. The concentrations of the compounds in the samples were determined by LC-MS / MS with a limit of quantitation of 1 ng / mL. The pharmacokinetic parameters of the compounds were calculated using the WinNolin non-compartmental model.

[0429] result

[0430] After intravenous administration, the in vivo exposure levels (AUC) of A1 and A2 were measured. last The effective clearances were 881 hr*ng / mL and 949 hr*ng / mL, respectively, with average total clearances (CL) of 18.9 mL / min / kg and 16.7 mL / min / kg, respectively.

[0431] After oral administration, the in vivo exposure levels (AUC) of A1 and A2 last The bioavailability of the cynomolgus monkeys was 388 ng*hr / mL and 1758 ng*hr / mL, respectively. Compared with the intravenous data, the oral bioavailability was 4.61% and 18.8%, respectively.

[0432] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0433] The foregoing embodiments and methods described in this invention may vary based on the capabilities, experience, and preferences of those skilled in the art.

[0434] Listing the steps of the method in a certain order in this invention does not constitute any restriction on the order of the method steps.

Claims

1. A compound having the following structure: （Ⅱ） in: Ring E is selected from: ; In ring E, each R0 is independently selected from: -H, C 1-10 Straight-chain / branched alkyl, -N(C) 0-10 Alkyl)(C 0-10 alkyl), -OC 0-10 Alkyl or C 3-10 cycloalkyl; R1 is selected from: ； R2 is -NH2; R3 is selected from: -H, halogen, -OC 0-10 Alkyl, C 1-10 Straight-chain / branched alkyl groups; R4 is -C≡CR 10 ; R5, R6, and R7 are independently selected from: -H, halogen, -CN, and C. 1-3 Straight-chain / branched alkyl, -OC 0-3 Alkyl, N-containing C 1-3 Straight-chain / branched alkyl groups; R8 and R9 are independently selected from: -H, C 1-3 Straight-chain / branched alkyl groups; R 10 for ; R 11 R 12 Independently selected from: -H, -CF3, -CHF2, -CH2F, C 1-10 Straight-chain / branched alkyl, C 3-10 cycloalkyl, or R 11 R 12 With R 11 and R 12 The carbon atoms between them form C 3-8 cycloalkyl; Or its pharmaceutically acceptable salt; The alkyl group of C0 is H.

2. The compound according to claim 1, characterized in that, R1 is .

3. The compound according to claim 1, characterized in that, R 11 and R 12 Independently selected from: -H, -CF3, -CHF2, -CH2F, -CH3, -CH2CH3, , or R 11 R 12 With R 11 and R 12 The formation of carbon atoms between .

4. The compound according to claim 1, characterized in that, R 11 and R 12 Independently selected from: -H, -CF3, -CHF2, -CH2F, -CH3, -CH2CH3, .

5. The compound according to claim 1, characterized in that, Each R0 is independently selected from: -H, -CH3, -CH2CH3 or -NH2.

6. The compound according to claim 1, characterized in that, R3 is selected from: -H, -F, -OCH3.

7. The compound according to claim 1, characterized in that, R5, R6, and R7 are independently selected from: -H, -F, -Cl, -CH3, -OCH3, -CH2NH2, -CH2N(CH3)2, and -CN.

8. The compound according to claim 1, characterized in that, R8 and R9 are independently selected from: -H, -CH3.

9. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is selected from the following structures: 。 10. A crystal form of 4-(3-(((2-amino-5-(1-(1-trideuterated methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yne-2-ol, characterized in that, The crystal form has an XRPD pattern that is essentially as shown in Figure 1.

11. A crystal form of 4-(3-(((2-amino-5-(1-(1-trideuterated methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol, characterized in that, The crystal form has an XRPD pattern that is essentially as shown in Figure 6.

12. A crystal form of 4-(3-(((2-amino-5-(1-(1-trideuterated methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yne-2-ol hydrochloride, characterized in that, The crystal form has an XRPD pattern that is essentially as shown in Figure 11.

13. A pharmaceutical composition comprising the compound of any one of claims 1-9 or a pharmaceutically acceptable salt thereof, or the crystal form of any one of claims 10-12, and a pharmaceutically acceptable excipient.

14. The use of the compound of any one of claims 1-9 or a pharmaceutically acceptable salt thereof, or the crystal form of any one of claims 10-12, or the pharmaceutical composition of claim 13 in the preparation of a medicament for the prevention and / or treatment of HPK1 kinase-mediated tumors; The tumors are selected from: lymphoma, sarcoma, mesothelioma, schwannoma, meningioma, melanoma, lymphoid malignant tumors, lung cancer, hepatocellular carcinoma, gastric cancer, intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, breast cancer, uterine cancer, kidney cancer, prostate cancer, esophageal cancer, biliary tract tumors, head and neck cancers, and hematologic malignancies.

15. The application as described in claim 14, characterized in that, The sarcoma mentioned is either liposarcoma or synovial cell sarcoma.

16. The application as described in claim 14, characterized in that, The lung cancer mentioned is squamous cell carcinoma.

17. The application as described in claim 16, characterized in that, The squamous cell carcinoma mentioned is epithelial squamous cell carcinoma.

18. The application as described in claim 14, characterized in that, The lung cancer mentioned is either small cell lung cancer or non-small cell lung cancer.

19. The application as described in claim 18, characterized in that, The non-small cell lung cancer mentioned is squamous cell carcinoma of the lung.

20. The application as described in claim 14, characterized in that, The lung cancer mentioned is adenocarcinoma.

21. The application as described in claim 14, characterized in that, The breast cancer mentioned is metastatic breast cancer.

22. The application as described in claim 14, characterized in that, The colorectal cancer mentioned refers to colon cancer, rectal cancer, or colorectal cancer.

23. The application as described in claim 14, characterized in that, The aforementioned hematologic malignancy is leukemia.

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