Deuterated HPK1 kinase inhibitors and their preparation and use

Abstract

The present invention provides a compound of general formula I or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof, and a method for preparing and using the same, and particularly provides the use of a compound of general formula I or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof as an HPF1 kinase inhibitor for the prevention and / or treatment of tumors, diseases caused by pathogen infection, or diseases associated with pathogen infection. The compound of general formula I contains at least one deuterium atom, which has high stability and high bioavailability in the animal body.

Description

[Technical Field]

[0001] The present invention relates to the pharmaceutical technology field, specifically to a deuterated HPK1 kinase inhibitor, and a method for preparing and using the same. The present invention also relates to a crystalline form of the deuterated HPK1 kinase inhibitor (particularly, 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol and salts thereof), and a method for preparing and using the same. [Background technology]

[0002] HPK1 kinase is involved in many signal cascades, including growth factor signaling, MAPK signaling, cytokine signaling, apoptosis signaling, growth factor signaling, antigen receptor signaling, etc. HPK1 kinase is an important functional activator of the JNK / SAPK signaling pathway, and upon activation, it can selectively activate the MAPK signaling pathway of c-Jun N-terminal kinase (JNK).

[0003] HPK1 kinase can be a target for immunotherapy. It is activated by lymphocyte antigen receptors and inhibits AP-1, which acts in tumor formation and progression by promoting cell proliferation, suppressing differentiation, and promoting tumor cell invasion and metastasis. Targeted disruption of HPK1 kinase alleles can increase Th1 cytokine production by T cells in TCR responses.

[0004] Sawasdikosol (HPK1 as a novel target for cancer immunotherapy, Immunol Res, 54 (2012), pp. 262-265) reported that HPK1 kinase- / - T cells proliferate faster than their monomeric wild-type counterparts and are resistant to prostaglandin E2 (PGE2)-mediated suppression. Most notably, mice transfected with HPK1 kinase- / - T cells exhibit resistance to cancer tumor growth, and HPK1 kinase-deficient dendritic cells (DCs) are highly antigenic, enabling HPK1 kinase- / - DCs to elicit a better antitumor immune response as an antitumor vaccine. Inhibition of HPK1 kinase activity with a small molecule inhibitor can activate the enhanced antitumor activity of these two types of cells, ultimately synergistically amplifying their potential antitumor capabilities. At the same time, because the transfected HPK1 kinase is not expressed in major organs, it is predicted that inhibitors of HPK1 kinase activity will not cause serious complications.

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

[0006] It is clear that HPK1 kinase plays an important role in disease treatment, particularly cancer treatment, and there is currently an urgent need to develop small molecule inhibitors of HPK1 kinase.Patent document CN110396087A discloses heterocyclic compounds that can be used as HPK1 kinase inhibitors. Summary of the Invention

[0007] The present invention provides compounds of general formula I:

[0008] [ka] During the ceremony, A is CR 10 or selected from N; Q is selected from O or S; x and z are independently selected from integers 0 to 6 (e.g., 0, 1, 2, 3, 4, 5, 6); y is 0 or 1; Ar is selected from an aromatic 5-membered heterocyclic group, an aromatic 6-membered heterocyclic group, or phenyl, wherein the aromatic 5-membered heterocyclic group is selected from furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, or selenothiazolyl, and the aromatic 6-membered heterocyclic group is selected from pyridyl, pyridazinyl, pyrimidinyl, or pyrazinyl, and optionally, H in the aromatic 5-membered heterocyclic group, the aromatic 6-membered heterocyclic group, or the phenyl is selected from -D, -SO2, -SON(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, C 1~10 Straight / branched alkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), -OC 0~10 Alkyl, C 3~10 It may be substituted with cycloalkyl, -Oheterocycloalkyl, -Nheterocycloalkyl, -Nheterocycloaryl, -Oheterocycloaryl or -Sheterocycloaryl, provided that the alkyl portion is not substituted with -SO, -SON(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), -COO(C 0~10 alkyl), -Nheterocycloaryl, -Oheterocycloaryl or -Sheterocycloaryl, provided that one or more H atoms to which a C atom or heteroatom is attached may be replaced with deuterium; R2 is -H, -D, halogen, -NO2, -CN, C 1~10 Straight / branched alkyl, C 3~10 Cycloalkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), -CF3, -OCF3, -OCHF2, -OCH2F or -OC 0~10 alkyl, provided that one or more H atoms to which the C atom is attached may be replaced by deuterium; 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, R7 is absent); When present, R3, R4, R5, R6, and R7 are independently -H, -D, halogen, -CN, -OC 0~10 Alkyl, -CO(C 0~10 alkyl), -CON(C 0~10 Alkyl)(C 0~10 alkyl), C 1~10 Straight chain / branched alkyl, O- or N-containing heteroalkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), C 3~10 Cycloalkyl, -C≡CR 10, -Oheterocycloalkyl, -Nheterocycloalkyl, or R5 and R4, R4 and R3, R3 and R7, R7 and R6 together with the carbon atoms therebetween are selected from C 3~8 Cycloalkyl or C containing -O-, -S- 3~8 Heterocycloalkyl, -Nheteroaryl, -Oheteroaryl or -Sheteroaryl, phenyl, provided that the alkyl portion is not -SO, -SON(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, C 1~10 Straight / branched alkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), -OC 0~10 Alkyl, C 3~10 optionally substituted with one or more of cycloalkyl, -Oheterocycloalkyl, -Nheterocycloalkyl, -Nheterocycloaryl, -Oheterocycloaryl or -Sheterocycloaryl, provided that one or more H atoms to which a C atom or heteroatom is attached may be replaced with deuterium; R8 and R9 are independently -H, -D, halogen, C 1-10 selected from straight chain / branched alkyl, provided that one or more H atoms to which the C atom is attached may be replaced by deuterium; R 10 H, -D, C 1~5 Straight / branched alkyl, C 3~10 cycloalkyl,

[0009] [ka] wherein one or more H atoms to which the C atom is attached may be replaced by deuterium; R 11 , R 12 are independently -H, -D, -CF3, -CHF2H, -CH2F, C 1-10 Straight / branched alkyl, -CH=C(C 0~10 Alkyl)(C 0~10 alkyl), -C≡C(C 0~10 alkyl), C 3~10 cycloalkyl, an aromatic five-membered ring group, or an aromatic six-membered ring group; or R 11 , R 12 , and R 11 and R 12 The carbon atom between 3~8 Cycloalkyl or C containing -O-, -S- 3~8 Heterocycloalkyl, C 4~9 Fused cycloalkyl, C 5~10 Spirocycloalkyl, C 4~9 Bridged cycloalkyl, C 3~7 Cyclolactam, C 3~7 Cyclolactone, C 3~7 Forms a cycloketone, provided that the alkyl moiety is -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 0-10optionally substituted with one or more of alkyl, -Nheterocycloaryl, -Oheterocycloaryl, or -Sheterocycloaryl, provided that one or more H atoms to which a C atom or heteroatom is attached may be replaced with deuterium; And the compound of general formula I contains at least one deuterium atom.

[0010] In one embodiment of the present invention, A is CR 10 and especially CH.

[0011] In one embodiment of the present invention, Q is O.

[0012] In one embodiment of the present invention, x is 0.

[0013] In one embodiment of the present invention, z is 1.

[0014] In one embodiment of the present invention, y is one.

[0015] In one embodiment of the present invention, B1, B2, B3, B4 and B5 are all C, i.e. in general formula I:

[0016] [ka] teeth,

[0017] [ka] is.

[0018] In another embodiment of the present invention, at least one of B1, B2, B3, B4 and B5 is N.

[0019] Specifically, B2 is C, and at least one of B1, B3, B4, and B5 is N.

[0020] More specifically, B2 is C and B1 is N.

[0021] More specifically, B2 is C and B3 is N.

[0022] More specifically, B2 is C and B4 is N.

[0023] More specifically, B2 is C and B5 is N.

[0024] More specifically, B2 is C and B3 and B4 are N, or both B3 and B5 are N.

[0025] Specifically, Ar is selected from thiazolyl, selenothiazolyl, imidazolyl, pyrazolyl, and pyridyl.

[0026] In one embodiment of the present invention, the compound of general formula I has the following structure:

[0027] [ka] However, the E ring is

[0028] [ka] Selected from; In the E ring, each R is independently -H, -D, C 1~10 Straight / 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; provided that H attached to the C atom or heteroatom may be replaced by deuterium; R1 is -H, -D, -O heterocycloalkyl, -N heterocycloalkyl, C 1~10 Straight / branched alkyl, C 3~10 Cycloalkyl, -OC 0~10 Alkyl, -N(C0~10 Alkyl)(C 0~10 alkyl), -SO2(C 0~10 alkyl), -CO(C 0~10 alkyl), -O-phenyl, -S(C 0~10 alkyl), -Nheterocycloaryl, -Oheterocycloaryl or -Sheterocycloaryl; provided that H bonded to the C atom or heteroatom may be replaced by deuterium; R 2-9 has the corresponding definition above in the present invention; And R 0-9 At least one of the groups contains a deuterium atom.

[0029] 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, but R 2-9 does not contain deuterium atoms.

[0030] In another embodiment of the present invention, in general formula II, R2 contains at least one deuterium atom.

[0031] In another embodiment of the present invention, in general formula II, R3 contains at least one deuterium atom.

[0032] In another embodiment of the present invention, in general formula II, R4 contains at least one deuterium atom.

[0033] In another embodiment of the present invention, in general formula II, R5 contains at least one deuterium atom.

[0034] In another embodiment of the present invention, in general formula II, R6 contains at least one deuterium atom.

[0035] In another embodiment of the present invention, in general formula II, R8 and / or R9 contain at least one deuterium atom.

[0036] Specifically, each R0 independently represents C 1~5 Straight chain / branched alkyl or -N(C 0~10 Alkyl)(C 0~10 alkyl), provided that H attached to the C atom may be replaced with deuterium.

[0037] More specifically, each R0 is independently selected from -H, -D, -CH3, -CH2CH3, or -NH2.

[0038] Specifically, R1 is -Oheterocycloalkyl or -Nheterocycloalkyl, -SO2(C 0~3 alkyl), -O-phenyl, -S(C 0~4 alkyl), C 3~6 Cycloalkyl or C 3~5 It is selected from straight chain / branched alkyl, provided that H attached to a C atom or heteroatom may be replaced with deuterium.

[0039] More specifically, R1 is

[0040] [ka] , -CH3,

[0041] [ka] wherein H bonded to a C atom or N atom may be replaced with deuterium.

[0042] More specifically, R1 is

[0043] [ka] , -CH2D, -CHD2, -CD3,

[0044] [ka] is selected from.

[0045] Specifically, when R0 is adjacent to R1, R0, R1, and the carbon atom between them are C 3-8 Cycloalkyl or C containing -O-, -S- 3-8 Forms heterocycloalkyl, -Nheterocycloaryl, -Oheterocycloaryl or -Sheterocycloaryl, phenyl.

[0046] Specifically, R2 is -H, -D, halogen, -NO2, -CN, C 1~5 Straight / branched alkyl, C 3~10 Cycloalkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), -CF3, -OCF3, -OCHF2, -OCH2F or -OC 0~10 alkyl, provided that H bonded to a C atom or N atom may be replaced with deuterium.

[0047] More specifically, R2 is -NO2, -N(C 0~10 Alkyl)(C 0~10 alkyl), -OC 0~10 alkyl, and -OCF3, provided that H bonded to a C atom or N atom may be replaced by deuterium.

[0048] More specifically, R2 is selected from -NH2, -NHD, -ND2, or -NO2.

[0049] Specifically, R3 is -H, -D, halogen, or -OC. 0~10 Alkyl, -CO(C 0~10 alkyl), C 1~10 Straight / branched alkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl) or C 3~10 cycloalkyl, provided that the H attached to the C atom may be replaced by deuterium.

[0050] More specifically, R3 is -H, -D, halogen, -OC 0~10 Alkyl, C 1~10 It is selected from straight chain / branched alkyl, provided that H attached to the C atom may be replaced with deuterium.

[0051] More specifically, R3 is selected from -H, -D, -F, -OCH3, -OCH2D, -OCHD2, and -OCD3.

[0052] Specifically, R4 is -H, -D, halogen, or -OC. 0~10 Alkyl, -CO(C 0~10 alkyl), -CN, C 3~10 Cycloalkyl, -C≡CR 10 , C 1~10 Straight / branched alkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), -Oheterocycloalkyl or -Nheterocycloalkyl, provided that H bonded to a C atom or N atom may be replaced by deuterium.

[0053] More specifically, R4 is -H, -D, halogen, -OC 0~10 Alkyl, -CN, C 3~10 Cycloalkyl or -C≡CR 10 wherein H attached to the C atom may be replaced by deuterium. 10 The H bonded to the C atom of may be replaced by deuterium.

[0054] In one embodiment of the present invention, R4 is -H, -D, -F, -Cl, -OCH3, -OCH2D, -OCHD2, -OCD3, -CN,

[0055] [ka] or -C≡CR 10 is selected from.

[0056] Specifically, R5, R6, and R7 are independently -H, -D, halogen, -CN, or -OC. 0~10 Alkyl, -CO(C 0~10 alkyl), C 1~10 Straight / branched alkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), C 3~10 Cycloalkyl, -C≡CR 10 , -Oheterocycloalkyl or -Nheterocycloalkyl, C containing O or N 1~5 or R6, R7, and the carbon atom between R6 and R7 are selected from C 3~8 Cycloalkyl or C containing -O-, -S- 3~8 Forms a heterocycloalkyl, provided that H attached to a C atom or heteroatom may be replaced with deuterium.

[0057] More specifically, R5, R6, and R7 are independently -H, -D, halogen, -CN, C 1~3 Straight / branched alkyl, -OC 0~3 Alkyl, -CO(C 0~3 alkyl), N-containing C 1~3 R6, R7, and the carbon atoms between R6 and R7 are selected from linear / branched alkyl, or 3~8 Cycloalkyl or -O-containing C 3~8 Forms a heterocycloalkyl, provided that H bonded to a C atom or an N atom may be replaced with deuterium.

[0058] More specifically, R5, R6, and R7 are independently selected from -H, -D, -F, -Cl, -CH3, -CH2NH2, -CH2NH(CH3), -CH2N(CH3)2, -CN, -OCH3, and -COCH3; or R6, R7, and the carbon atoms between R6 and R7 form an -O-containing 5-membered cycloalkyl ring, with the proviso that the H bonded to the C atom or N atom may be replaced with deuterium.

[0059] 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).

[0060] 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).

[0061] 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).

[0062] In one embodiment of the present invention, R 10 teeth,

[0063] [ka] In one embodiment of the present invention, R 10 The H bonded to the C atom of may be replaced by deuterium.

[0064] Specifically, R 11 and R 12 are independently -H, -D, -CF3, -CHF2H, -CH2F, C 1~10 Straight / branched alkyl, -CH=C(C 0~10 Alkyl)(C 0~10 alkyl), C 3~10 cycloalkyl or aromatic 6-membered ring group, or R 11 , R 12and R 11 and R 12 The carbon atom between 3~8 Cycloalkyl, C 4~7 Fused cycloalkyl, C 5~9 Spirocycloalkyl, C 4~9 Bridged cycloalkyl, C 3~7 Cyclolactam, C 3~7 Cyclolactone, C 3~7 Forms a cycloketone, provided that the H on the C atom is -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, C 1~10 Straight / branched alkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), -OC 0~10 Alkyl, -CO(C 0~10 alkyl), C 3~10 and -S-heterocycloaryl, ... 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 0~10Alkyl)(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), -Nheterocycloaryl, -Oheterocycloaryl, or -Sheterocycloaryl may be optionally substituted with one or more of the following: -C atom or heteroatom; -Nheterocycloaryl, -Oheterocycloaryl, or -Sheterocycloaryl; and H bonded to a C atom or heteroatom may be replaced with deuterium.

[0065] More specifically, R 11 and R 12 are independently -H, -D, -CF3, -CHF2, -CH2F, C 1~5 Straight / branched alkyl, -CH=CH(C 0~10 alkyl), C 3~10 cycloalkyl or aromatic 6-membered ring group, or R 11 , R 12 and R 11 and R 12 The carbon atom between 3~6 Cycloalkyl, C 4~6 Fused cycloalkyl, C 5~8 Spirocycloalkyl, C 4~8 Bridged cycloalkyl, C 3~7 Cyclolactam, C 3~7 Cyclolactone, C 3~7 Forms a cycloketone, provided that the alkyl moiety is -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, C1~10 Straight / branched alkyl, -N(C 0~10 Alkyl)(C 0~10 alkyl), -OC 0~10 Alkyl, -CO(C 0~10 alkyl), C 3~10 and -S-heterocycloaryl, ... 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 It may be optionally substituted with one or more of -alkyl), -Nheterocycloaryl, -Oheterocycloaryl, or -Sheterocycloaryl, provided that H bonded to a C atom or heteroatom may be replaced with deuterium.

[0066] More specifically, R 11 and R 12 are independently -H, -D, -CF3, -CHF2, -CH2F, -CH3, -CH2CH3, -CH=CH2,

[0067] [ka] or R 11 , R 12 and R11 and R 12 The carbon atoms between

[0068] [ka] wherein H bonded to a C atom or N atom may be replaced with deuterium.

[0069] More specifically, R 11 and R 12 are independently -H, -D, -CF3, -CHF2, -CDF2, -CH2F, -CD2F, -CH3, -CH2D, -CHD2, -CD3, -CH2CH3, -CH2CD3,

[0070] [ka] is selected from.

[0071] Specifically, R8 and R9 are independently -H, -D, C 1~10 It is selected from straight chain / branched alkyl, provided that H attached to the C atom may be replaced with deuterium.

[0072] More specifically, R8 and R9 are independently -H, -D, C 1~3 It is selected from straight chain / branched alkyl, provided that H attached to the C atom may be replaced with deuterium.

[0073] More specifically, R8 and R9 are independently selected from -H, -D, -CH3, -CH2D, -CHD2, and -CD3.

[0074] Specifically, compounds of general formula I have the following structure:

[0075] (1)

[0076] [ka]

[0077]

change

[0078]

change

[0079]

change

[0080]

change

[0081] (2)

[0082]

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[0083]

change

[0084]

change

[0085]

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[0086]

change

[0087] (3)

[0088]

change

[0089]

change

[0090]

change

[0091]

change

[0092] (4)

[0093]

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[0094]

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[0095]

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[0096]

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[0097] (5)

[0098]

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[0099]

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[0100] [ka]

[0101] Specifically, in the above compounds, any atom not specified as deuterium is present in its natural isotopic abundance.

[0102] Specifically, in the above compound, at least (for example) 95% of the positions defined as "heavy hydrogen" are doped with deuterium.

[0103] The present invention provides a method for preparing compounds of general formula I, comprising the following steps:

[0104] (1)

[0105] [ka] undergoes a condensation reaction

[0106] [ka] Generate R 13 is a halogen or

[0107] [ka] Selected from R 14 is selected from -OH or -F.

[0108] (2)

[0109] [ka] undergoes a condensation reaction

[0110] [ka] Generate R 15is selected from -Br or -SnBu3.

[0111] Specifically, R in step (1) above 13 is -Br.

[0112] Specifically, R in step (2) above 13 -Br or

[0113] [ka] and R 13 When is -Br, R 15 is -SnBu3, and R 13 but

[0114] [ka] When R 15 is -Br.

[0115] The present invention further provides pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates of the compounds of general formula I above.

[0116] Specifically, the pharmaceutically acceptable salts include acid addition salts and base addition salts.

[0117] Specifically, the acid addition salts include, but are not limited to, salts derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphonic acid, and salts derived from organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, and aliphatic and aromatic sulfonic acids. Thus, these salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, hydrochloride, hydrobromide, iodide, acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, and methanesulfonate salts, as well as salts of amino acids such as arginate, gluconate, galacturonate, and the like. Acid addition salts can be prepared by contacting the free base with a sufficient amount of the desired acid in a conventional manner to form the salt. The free base can be regenerated by contacting the salt with a base and separated in a conventional manner.

[0118] Specifically, the base addition salts are formed with metals or amines, such as alkali metal or alkaline earth metal hydroxides, or organic amines. Examples of metal cations include, but are not limited to, sodium, potassium, magnesium, and calcium. Examples of suitable amines include, but are not limited to, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine (ethane-1,2-diamine), N-methylglucamine, and procaine. Base addition salts can be prepared by contacting the free acid with a sufficient amount of the desired base in a conventional manner to form the salt. The free acid can be regenerated by contacting the salt with an acid and isolated in a conventional manner.

[0119] In one embodiment of the invention, the pharmaceutically acceptable salt is a hydrochloride salt.

[0120] Specifically, the stereoisomers include enantiomers, diastereomers, and geometric isomers. Some compounds of the present invention have a cycloalkyl group that may be substituted at one or more carbon atoms, and in this case, all geometric forms, including cis and trans groups and mixtures thereof, are within the scope of the present invention.

[0121] Specifically, the solvate refers to a physical combination of the compound of the present invention with one or more solvent molecules. This physical combination includes varying degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, for example, when one or more solvent molecules are incorporated into the lattice of a crystalline solid, the solvate can be isolated. "Solvate" includes solution-phase and separable solvates. Representative solvates include ethanolates, methanolates, etc. "Hydrate" is a solvate in which one or more solvent molecules are HO.

[0122] Specifically, the prodrug refers to a form of the compound of Formula I suitable for administration to a patient that is effective for its intended use without undue toxicity, irritation, allergic response, etc., and includes acetal, ester, and zwitterionic forms. The precursor is converted in vivo (e.g., by hydrolysis in blood) to yield the parent compound of the formula.

[0123] The present invention further provides crystalline forms of the compounds of general formula I above and pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates thereof.

[0124] Specifically, the present invention provides a crystalline form of 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol, which has the following structure:

[0125] [ka]

[0126] Specifically, the crystalline form is crystalline form A, which has characteristic peaks (major characteristic diffraction peaks) at at least three (or all) of the following 2θ values ​​in an XRPD pattern: 13.1°±0.2°, 16.3°±0.2°, 17.5°±0.2°, and 23.8°±0.2°.

[0127] Specifically, the XRPD pattern of crystalline form A further has characteristic peaks (secondary characteristic diffraction peaks) at at least three (at least four, at least five, at least six, at least seven, or all) of the following 2θ values: 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°.

[0128] Specifically, the crystalline form A has an XRPD pattern essentially as shown in FIG.

[0129] Specifically, the DSC pattern of the above-mentioned crystalline form A has an endothermic peak at about 168.8°C.

[0130] Specifically, the crystalline form A has a DSC pattern essentially as shown in FIG.

[0131] Specifically, when the crystalline form A is heated from room temperature to 170°C, the weight loss is about 1.1%.

[0132] Specifically, the crystalline form A has a TGA pattern essentially as shown in FIG.

[0133] Specifically, the above crystalline form A is an anhydrous crystalline form.

[0134] Specifically, the crystalline form is crystalline form B, which has characteristic peaks (major characteristic diffraction peaks) at at least three (or all) of the following 2θ values ​​in an XRPD pattern: 5.7°±0.2°, 11.3°±0.2°, 22.7°±0.2°, and 23.5°±0.2°.

[0135] Specifically, the XRPD pattern of crystalline form B further has characteristic peaks (secondary characteristic diffraction peaks) at at least three (at least four, at least five, or all) of the following 2θ values: 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°.

[0136] Specifically, the crystalline form B has an XRPD pattern essentially as shown in FIG.

[0137] Specifically, the DSC pattern of the crystalline form B has at least one endothermic peak at about 59.5°C, 95.6°C, 150.8°C, and 160.9°C.

[0138] Specifically, the crystalline form B has a DSC pattern essentially as shown in FIG.

[0139] Specifically, when the crystalline form B is heated from room temperature to 70°C, the weight loss is about 13.2%, and when it is subsequently heated to 170°C, the weight loss is about 8.5%.

[0140] Specifically, the crystalline form B has a TGA pattern essentially as shown in FIG.

[0141] Specifically, the above crystalline form B is an EtOAc solvate.

[0142] Specifically, the present invention further provides a crystalline form of 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol hydrochloride.

[0143] Specifically, the crystalline form is crystalline form A, which has characteristic peaks (major characteristic diffraction peaks) at at least three (at least four, or all) of the following 2θ values ​​in an XRPD pattern: 13.0°±0.2°, 16.3°±0.2°, 17.5°±0.2°, 19.4°±0.2°, and 23.8°±0.2°.

[0144] Specifically, the XRPD pattern of crystalline form A further has characteristic peaks (second-order characteristic diffraction peaks) at at least three (at least four, or all) of the following 2θ values: 8.1°±0.2°, 12.1°±0.2°, 15.3°±0.2°, 18.0°±0.2°, and 21.4°±0.2°.

[0145] Specifically, the crystalline form A has an XRPD pattern essentially as shown in FIG.

[0146] Specifically, the DSC pattern of the above-mentioned crystalline form A has endothermic peaks at about 81.9°C and about 156.0°C.

[0147] Specifically, the crystalline form A has a DSC pattern essentially as shown in FIG.

[0148] Specifically, when the crystalline form A is heated from room temperature to 150°C, the weight loss is about 8.6%.

[0149] Specifically, the crystalline form A has a TGA pattern essentially as shown in FIG.

[0150] The present invention further provides methods for preparing the above crystalline forms.

[0151] Specifically, the production method is one or a combination of methods selected from the group consisting of anti-solvent addition, anti-anti-solvent addition, gas-solid diffusion, room temperature suspension and stirring, 5°C suspension and stirring, slow evaporation, slow cooling, gas-liquid diffusion, and high polymer derivatization.

[0152] Specifically, the poor solvent addition method involves dissolving the target product in a good solvent and then adding a poor solvent to the resulting solution (which is then, for example, evaporated at room temperature to obtain a solid, or stirred at −20° C. to obtain a solid).

[0153] Specifically, the anti-solvent reverse addition method involves dissolving the target product in a good solvent and then adding the resulting solution to a anti-solvent (which is then, for example, evaporated at room temperature to obtain a solid, or stirred at −20° C. to obtain a solid).

[0154] 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 involve dissolving the target product in a solvent and then drying it to obtain a solid.

[0155] Specifically, the high polymer derivatization method involves dissolving the target product in a solvent, adding a high polymer, and volatilizing it at room temperature to obtain a solid.

[0156] Specifically, the method for preparing the crystalline form A of 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol is one or a combination of methods selected from the group consisting of a poor solvent addition method, a poor solvent reverse addition method, a gas-solid diffusion method, a room temperature suspension stirring method, a 5°C suspension stirring method, a slow evaporation method, a slow cooling method, a gas-liquid diffusion method, and a high polymer derivatization method.

[0157] Specifically, in preparing the above-mentioned Crystalline Form A, in the anti-solvent addition method, the good solvent may be selected from MeOH, acetone, DMSO, EtOAc, EtOH, DCM, CHCl3, THF, IPA, ACN, and 1,4-dioxane, and the anti-solvent may be selected from MTBE, toluene, n-heptane, and water.

[0158] In one embodiment of the present invention, in the anti-solvent addition method, the good solvent may be selected from MeOH, acetone, DMSO, and the anti-solvent is MTBE.

[0159] In another embodiment of the present invention, in the anti-solvent addition method, the good solvent may be selected from EtOH, DCM, and the anti-solvent is toluene.

[0160] In another embodiment of the present invention, in the anti-solvent addition method, the good solvent may be selected from CHCl3, THF, IPA, and the anti-solvent is n-heptane.

[0161] In another embodiment of the present invention, in the anti-solvent addition method, the good solvent may be selected from acetone, ACN, 1,4-dioxane, and the anti-solvent is water.

[0162] Specifically, in preparing the above-mentioned crystalline form A, in the anti-solvent reverse addition method, the good solvent may be selected from MeOH, MIBK, acetone, anisole, EtOH, THF, EtOAc, and DCM, and the anti-solvent may be selected from toluene, n-heptane, water, and MTBE.

[0163] In one embodiment of the present invention, in the anti-solvent reverse addition method, the good solvent may be MIBK and the anti-solvent may be toluene.

[0164] In another embodiment of the present invention, in the anti-solvent reverse addition method, the good solvent may be selected from acetone and anisole, and the anti-solvent may be n-heptane.

[0165] In another embodiment of the present invention, in the anti-solvent reverse addition method, the good solvent may be selected from EtOH, THF, and the anti-solvent may be water.

[0166] In another embodiment of the present invention, in the anti-solvent reverse addition method, the good solvent may be selected from EtOAc, DCM, and the anti-solvent may be MTBE.

[0167] Specifically, for preparing the above crystalline form A, in the gas-solid diffusion method, the solvent may be selected from water, DCM, EtOH, MeOH, ACN, THF, CHCl3, acetone, DMSO, EtOAc, 1,4-dioxane, and IPA.

[0168] Specifically, in preparing the above-mentioned crystalline form A, in the room temperature suspension stirring method, the solvent may be selected from MTBE, IPAc, n-heptane, toluene, water, EtOH / toluene (e.g., 1:3 ratio, v / v), DMSO / MTBE (e.g., 1:4 ratio, v / v), acetone / water (e.g., 1:4 ratio, v / v), IPA / n-heptane (e.g., 1:4 ratio, v / v), EtOAc / n-heptane (e.g., 1:4 ratio, v / v), anisole / toluene (e.g., 1:4 ratio, v / v), DMAc / water (e.g., 1:4 ratio, v / v), and THF / water (e.g., 1:4 ratio, v / v).

[0169] Specifically, in the case of producing the above-mentioned crystalline form A by a suspension stirring method at 5°C, the solvent may be selected from MTBE, toluene, water, IPA / n-heptane (e.g., 1:2 ratio, v / v), MEK / n-heptane (e.g., 1:2 ratio, v / v), EtOAc / toluene (e.g., 1:2 ratio, v / v), CPME / toluene (e.g., 1:2 ratio, v / v), NMP / water (e.g., 1:4 ratio, v / v), THF / water (e.g., 1:4 ratio, v / v), ACN / water (e.g., 1:2 ratio, v / v), IPAc / DCM (e.g., 1:1 ratio, v / v), MeOH / toluene (e.g., 1:4 ratio, v / v), DCM / MTBE (e.g., 1:4 ratio, v / v), and THF / n-heptane (e.g., 1:4 ratio, v / v).

[0170] Specifically, for the preparation of the above crystalline form A, in the slow evaporation method, the solvent may be selected from EtOH, acetone, IPAc, THF, CPME, anisole, ACN / water (e.g., 9:1 ratio, v / v), MeOH / DCM (e.g., 1:1 ratio, v / v), acetone / EtOAc (e.g., 2:1 ratio, v / v), and THF / water (e.g., 4:1 ratio, v / v).

[0171] Specifically, in the slow cooling method for producing the above-mentioned crystalline form A, the solvent may be selected from CPME, toluene, ACN / toluene (e.g., 1:2 ratio, v / v), acetone / n-heptane (e.g., 1:1 ratio, v / v), THF / toluene (e.g., 1:2 ratio, v / v), MeOH / water (e.g., 1:1 ratio, v / v), and CHCl3 / MTBE (e.g., 1:1 ratio, v / v).

[0172] Specifically, in the production of the above-mentioned crystalline form A, in the gas-liquid diffusion method, the good solvent may be selected from EtOH, THF, and DMSO, and the poor solvent may be selected from n-heptane, MTBE, toluene, cyclohexane, and water.

[0173] In one embodiment of the present invention, in the gas-liquid diffusion method, the good solvent may be EtOH, and the poor solvent may be selected from n-heptane, MTBE, and toluene.

[0174] In another embodiment of the present invention, in the gas-liquid diffusion method, the good solvent may be THF, and the poor solvent may be selected from n-heptane, cyclohexane, and MTBE.

[0175] In another embodiment of the present invention, in the gas-liquid diffusion method, the good solvent may be DMSO, and the poor solvent may be selected from toluene, MTBE, and water.

[0176] Specifically, in the preparation of the above-mentioned crystalline form A, in the high polymer derivatization method, the solvent may be selected from MEK, ACN / toluene (e.g., 4:1 ratio, v / v), THF / water (e.g., 9:1 ratio, v / v), EtOAc, acetone / 2-MeTHF (e.g., 1:1 ratio, v / v), and MeOH / DCM (e.g., 1:1 ratio, v / v), and the high polymer may be one or a combination selected from polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropylmethylcellulose, methylcellulose, polycaprolactone, polyethylene glycol, polymethylmethacrylate, sodium alginate, and hydroxyethylcellulose.

[0177] In one embodiment of the present invention, in the high polymer derivatization method, the high polymer is a mixture of polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl chloride, polyvinyl acetate, hydroxypropyl methylcellulose and methylcellulose (e.g., in a 1:1:1:1:1:1 mass ratio), and the solvent may 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).

[0178] In another embodiment of the present invention, in the high polymer derivatization method, the high polymer is a mixture of polycaprolactone, polyethylene glycol, polymethyl methacrylate, sodium alginate and hydroxyethyl cellulose (e.g., in a 1:1:1:1:1:1 mass ratio), and the solvent may be selected from EtOAc, acetone / 2-MeTHF (e.g., in a 1:1 ratio, v / v).

[0179] Specifically, the method for producing crystalline form B of 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol is the anti-solvent addition method.

[0180] In one embodiment of the present invention, for the preparation of crystalline form B, the good solvent is EtOAc and the poor solvent is toluene.

[0181] The present invention further provides a pharmaceutical composition comprising a compound of general formula I above, or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate, or crystalline form thereof, and a pharmaceutically acceptable excipient.

[0182] Specifically, the additive is one or more selected from a carrier, a diluent, a binder, a lubricant, a wetting agent, etc. Specifically, the pharmaceutical composition comprises a therapeutically effective amount of a compound of general formula I. In certain embodiments, these pharmaceutical compositions can be used to treat diseases or conditions mediated by HPK1 kinase.

[0183] Specifically, the pharmaceutical composition may be in the form of tablets (e.g., sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, buccal tablets, etc.), pills, powders, granules, capsules (e.g., soft capsules, microcapsules), tablets, syrup, emulsions, suspensions, controlled-release preparations (e.g., immediate-release preparations, sustained-release preparations, sustained-release microcapsules), aerosols, film preparations (e.g., orally disintegrating films, oral mucosal patches), injections (e.g., subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections), drip infusions, transdermal preparations, ointments, lotions, tapes, suppositories (e.g., rectal suppositories, vaginal suppositories), pellets, nasal drops, pulmonary preparations (inhalants), eye drops, etc.

[0184] Specifically, each dosage form of the pharmaceutical composition can be prepared according to a conventional manufacturing method in the pharmaceutical field, for example, by mixing an active ingredient with one or more additives and then preparing the mixture into a desired dosage form.

[0185] Specifically, the pharmaceutical composition may contain the active ingredient in a weight ratio of 0.1 to 99.5% (e.g., 0.1%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5%).

[0186] The present invention further provides the use of the compound of general formula I above and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the above crystalline forms, or the above pharmaceutical compositions, in the manufacture of a medicament for the prevention and / or treatment of tumors.

[0187] The present invention further provides the use of the compounds of general formula I above and their pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, and crystalline forms above in combination with PD-1, PD-L1, CTLA-4, TIM-3, TGF-β and its receptors, LAG3 antagonists or TLR4, TLR7, TLR8, TLR9, STING agonists in tumor immunotherapy.

[0188] The present invention further provides the use of the compounds of general formula I above and their pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, as well as the above crystalline forms, in combination with CAR-T immunotherapy in tumor immunotherapy.

[0189] Specifically, the above-mentioned CAR-T immunotherapy refers to chimeric antigen receptor T-cell immunotherapy, which is a type of cell therapy based on the basic principle of using the patient's own immune cells to remove cancer cells.

[0190] Specifically, the tumor is a malignant tumor, such as lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuroendocrine tumor, carcinoid tumor, gastrinoma, pancreatic islet cell tumor, mesothelioma, schwannoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or malignant lymphoma, squamous cell carcinoma, epithelial squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, Cancer, including, but not limited to, lung adenocarcinoma, lung squamous cell carcinoma, peritoneal cancer, 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 cancer, head and neck cancer, and malignant hematological tumors.

[0191] The present invention further provides use of the compound of general formula I and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the crystalline forms described above, or the pharmaceutical compositions described above, in the manufacture of a medicament for the prevention and / or treatment of a disease caused by or associated with pathogen infection.

[0192] Specifically, the pathogen may be a microorganism, a parasite (such as a protozoan or a helminth), or another vector. Specifically, the microorganism may be one or more selected from the group consisting of a virus, a chlamydia, a rickettsia, a mycoplasma, a bacterium, a spirochete, a fungus, and the like.

[0193] In one embodiment of the present invention, the pathogen is a virus, such as Adenoviridae (e.g., adenovirus), Herpesviridae (e.g., HSV1 (herpes labialis), HSV2 (genital herpes), VZV (chickenpox), EBV (Epstein-Barr virus), CMV (cytomegalovirus)), Poxviridae (e.g., smallpox virus, cowpox virus), Papovaviridae (e.g., human papillomavirus (HPV)), Parvoviridae (e.g., B19 virus), Hepadnaviridae (e.g., For example, hepatitis B virus (HBV), Polyomaviridae (e.g., polyomavirus), Reoviridae (e.g., reovirus, rotavirus), Picornaviridae (e.g., enterovirus, foot-and-mouth disease virus), Caliciviridae (e.g., Norwalk virus, hepatitis E virus), Togaviridae (e.g., rubella virus), Arenaviridae (e.g., lymphocytic choriomeningitis virus), Retroviridae (HIV), Flaviviridae (e.g., dengue virus, Zika virus, encephalitis B virus), virus, chikungunya virus, yellow fever virus, hepatitis C virus (HCV), West Nile virus, etc.), Orthomyxoviridae (e.g., influenza viruses (e.g., influenza A virus, influenza B virus, influenza C virus, etc.)), Paramyxoviridae (e.g., 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.), Bunyavirus Examples of such viruses include, but are not limited to, the family Mycoviridae (e.g., California encephalitis virus, Hantavirus), Rhabdoviridae (e.g., rabies virus), Filoviridae (e.g., Ebola virus, Marburg virus), Coronaviridae (e.g., HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, SARS-CoV-2, etc.), Astroviridae (e.g., astrovirus), and Borna disease virus (e.g., Borna disease virus).

[0194] Specifically, in the above use, the virus is HBV, HIV, HCV, HPV, Ebola virus, Marburg virus, influenza virus, parainfluenza virus, dengue virus, SARS-CoV, SARS-CoV-2, etc.

[0195] Specifically, the diseases caused by or associated with the above-mentioned pathogen infections include, but are not limited to, influenza, SARS, COVID-19, viral hepatitis (e.g., hepatitis B, hepatitis C, etc.), AIDS, dengue fever, Ebola virus disease, Marburg virus disease, etc.

[0196] The present invention further provides a method for preventing and / or treating tumors, which comprises administering an effective amount of the compound of the above general formula I of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the above crystalline forms, or the above pharmaceutical composition of the present invention to a subject in need thereof.

[0197] Specifically, tumors have the corresponding definitions set out above in the present invention.

[0198] The present invention further provides a method for the prevention and / or treatment of a disease caused by or associated with a pathogen infection, comprising the step of administering to a subject in need thereof an effective amount of the compound of the above general formula I of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs and solvates, or the above-mentioned crystalline forms, or the above-mentioned pharmaceutical composition of the present invention.

[0199] Specifically, pathogens and diseases have the corresponding definitions set forth above in the present invention. [Brief explanation of the drawings]

[0200] [Figure 1] 1 shows the XRPD pattern of the free base crystalline form A of Compound A2. [Figure 2] 1 shows the TGA / DSC pattern of the free base crystalline form A of Compound A2. [Figure 3] 1 shows the 1H NMR spectrum of the free base crystalline form A of Compound A2. [Figure 4] 1 shows an HPLC chromatogram of the free base crystalline form A of Compound A2. [Figure 5] 1 shows the XRPD patterns of the starting sample of the free base crystalline form A of Compound A2 and after standing in a closed space at 60° C. for 1 day. [Figure 6] 1 shows the XRPD pattern of the free base crystalline form B of Compound A2. [Figure 7] 1 shows the TGA / DSC pattern of the free base crystalline form B of Compound A2. [Figure 8] 1 shows the 1H NMR spectrum of the free base crystalline form B of Compound A2. [Figure 9] 1 shows the XRPD patterns of the free base crystalline form B of Compound A2 before and after standing at room temperature. [Figure 10] 1 shows the XRPD patterns of the free base crystalline form B of Compound A2 before and after nitrogen purging. [Figure 11] 1 shows the XRPD pattern of the hydrochloride salt of Compound A2. [Figure 12] 1 shows the TGA / DSC pattern of the hydrochloride salt of Compound A2. [Figure 13] 1 shows an overlay of the XRPD patterns of the free base crystalline form A of Compound A2 and the hydrochloride salt of Compound A2. DETAILED DESCRIPTION OF THE INVENTION

[0201] Unless otherwise defined, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0202] The term "C" in the present invention 0~10 Regarding "alkyl", CO alkyl refers to H, so C 0~10 Alkyl is H, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C 10 Contains alkyl.

[0203] The term "C" in the present invention 1~10 "Straight chain / branched alkyl" includes 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 Includes straight / branched alkyl.

[0204] The term "C" in the present invention 3~10 "Branched alkyl" includes isopropyl, isobutyl, tert-butyl, and isoamyl.

[0205] The term "C" in the present invention 3~10 "Cycloalkyl" includes C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, C8 cycloalkyl, C9 cycloalkyl, C 10 Includes cycloalkyl.

[0206] The term "C" in the present invention 3~8 "Cycloalkyl" includes C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, and C8 cycloalkyl.

[0207] The term "C" in the present invention 4~8 "Cycloalkyl" includes C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, and C8 cycloalkyl.

[0208] The term "C" in the present invention 4~6 "Cycloalkyl" includes C4 cycloalkyl, C5 cycloalkyl, and C6 cycloalkyl.

[0209] In the present invention, the term "halogen" includes fluorine, chlorine, bromine and iodine.

[0210] As used herein, the term "heterocycloalkyl" refers to a non-aromatic saturated monocyclic or polycyclic ring system containing 3 to 10 ring atoms, preferably 5 to 10 ring atoms, in which one or more ring atoms are not carbon atoms but are, for example, nitrogen, oxygen, or sulfur atoms. Preferred heterocycloalkyls contain 5 to 6 ring atoms. The prefix aza, oxa, or thia before heterocycloalkyl indicates that the system has at least one nitrogen, oxygen, or sulfur atom, respectively, as a ring atom.

[0211] As used herein, the term "heterocycloaryl" refers to an aromatic monocyclic or polycyclic ring system containing 5 to 14 ring atoms, preferably 5 to 10 ring atoms, in which one or more ring atoms are not carbon atoms, such as nitrogen, oxygen, or sulfur atoms. Preferred heterocycloaryls contain 5 to 6 ring atoms. Representative heterocycloaryls include pyrazinyl, furanyl, thienyl, pyridinyl, pyrimidinyl, isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,4-thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, 2,3-naphthyridinyl, imidazo[1,2-a]pyridine, imidazo[2,1-b]thiazolyl, benzofurazanyl, indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolyl, imidazolyl, thienopyridinyl, quinazolinyl, thienolimidinyl, pyrrolopyridinyl, imidazolopyridinyl, isoquinolyl, benzoazaindolyl, 1,2,4-triazinyl, benzothiazolyl, and the like.

[0212] In the present invention, "D" refers to deuterium, and "substituted with deuterium" refers to one or more hydrogen atoms being replaced with the corresponding number of deuterium atoms.

[0213] It should be recognized that there may be some variation in the abundance of natural isotopes in synthesized compounds depending on the source of the chemicals used in synthesis. Thus, the compounds of the present invention will inherently contain small amounts of deuterated isotopes. Despite such variations, the concentrations of stable hydrogen and carbon isotopes at natural abundance are still low and insignificant compared to the degree of stable isotope 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.

[0214] In the compounds of the present invention, atoms not specified as deuterium are present at their natural isotopic abundance. Unless otherwise specified, when a position is specifically designated as "H" or "hydrogen," it is to be understood that the position has hydrogen composed according to its natural isotopic abundance. Similarly, unless otherwise specified, when a position is specifically designated as "D" or "deuterium," it is to be understood that the position has deuterium at an abundance at least 3000 times greater than the natural abundance of deuterium (0.015%) (i.e., at least 45% deuterium doped).

[0215] As used herein, the term "isotopic enrichment factor" refers to the ratio between the abundance and natural abundance of a specified isotope.

[0216] In other embodiments, the isotopic enrichment factor for each designated deuterium atom of the compounds of the invention is at least 3500 (52.5% deuterium doped at each designated deuterium atom), at least 4000 (60% deuterium doped), at least 4500 (67.5% deuterium doped), at least 5000 (75% deuterium doped), at least 5500 (82.5% deuterium doped), at least 6000 (90% deuterium doped), at least 6333.3 (95% deuterium doped), at least 6466.7 (97% deuterium doped), at least 6600 (99% deuterium doped), or at least 6633.3 (99.5% deuterium doped).

[0217] The term "isotopes" refers to substances whose chemical structure differs from a specific compound of the invention only in isotopic composition.

[0218] The term "compound," when referring to a compound of the present invention, refers to a collection of molecules having the same chemical structure, except for possible isotopic variations between the constituent atoms of the molecule. Thus, it will be apparent to those skilled in the art that a compound represented by a specific chemical structure containing a designated deuterium atom will also contain small amounts of isotopes bearing hydrogen atoms at one or more of the designated deuterium positions in the structure. The relative amounts of such isotopes in a compound of the present invention will depend on various factors, such as the isotopic purity of the deuteration reagent used to prepare the compound and the efficiency of deuterium doping during each synthetic step used to prepare the compound. However, as noted 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.

[0219] Some of the abbreviations used in the present invention are explained below. XRPD: X-ray powder diffraction DSC: Differential scanning calorimetry TGA: Thermogravimetric analysis 1 H NMR: Liquid phase hydrogen nuclear magnetic resonance spectroscopy

[0220] In the present invention, the term "crystalline" is confirmed by characterizing the X-ray powder diffraction pattern. It is understood by those skilled in the art that the physicochemical properties discussed herein may be characterized with experimental error, which depends on instrument conditions, sample preparation, sample purity, and the like. In particular, it is well known to those skilled in the art that X-ray diffraction patterns typically vary depending on instrument conditions. It should be noted that the relative intensities of X-ray powder diffraction patterns may also vary depending on experimental conditions, and the order of peak intensities may not be the only or decisive factor. In fact, the relative intensities of diffraction peaks in an XRPD pattern are related to the preferred orientation of the crystals, and the peak intensities shown here are for illustrative purposes, not for absolute comparison. Furthermore, experimental errors in peak angles are typically less than 5%, and these angle errors must also be taken into account, typically allowing for an error of ±0.2°. Furthermore, experimental factors such as sample thickness may cause overall deviations in peak angles, and a certain degree of deviation is typically acceptable. Therefore, the X-ray powder diffraction pattern of the crystalline form of the present invention does not necessarily have to be completely identical to the X-ray powder diffraction pattern in the Examples herein. It is understood by those skilled in the art that the "same XRPD pattern" herein does not mean absolutely identical; the positions of the identical peaks may differ by ±0.2°, and some degree of variation in peak intensity is acceptable. Any crystalline form having a pattern identical or similar to the characteristic peaks of these patterns falls within the scope of the present invention. Those skilled in the art can compare the patterns listed in the present invention with the pattern of an unknown crystalline form to determine whether the two sets of patterns reflect the same crystalline form or different crystalline forms.

[0221] In one embodiment, crystalline form A of the present invention is pure and monolithic and is substantially free of any admixture of other crystalline forms. In the present invention, when used to refer to a new crystalline form, "substantially free" means that the crystalline form contains less than 20% by weight of other crystalline forms, particularly less than 10% by weight of other crystalline forms, further less than 5% by weight of other crystalline forms, and even less than 1% by weight of other crystalline forms.

[0222] It should be noted that the numerical values ​​and numerical ranges referred to in the present invention should not be construed narrowly as numerical values ​​or numerical ranges themselves. Those skilled in the art should understand that values ​​can vary around the specific numerical values ​​depending on the specific technical situation without departing from the concept and principles of the present invention. Such a variation range that can be predicted by those skilled in the art is often expressed as "about" in the present invention. When the term "about" appears before a numerical value of the present invention and refers to the numerical value, it means any value within a range of ±10%, preferably within a range of ±5%, more preferably within a range of ±2%, and preferably within a range of ±1% of the numerical value. For example, "about 10" should be interpreted as meaning 9 to 11, preferably 9.5 to 10.5, more preferably 9.8 to 10.2, and more preferably 9.9 to 10.1.

[0223] In the present invention, the term "room temperature" refers to the temperature of an item being close to or the same as the temperature of the space (e.g., the fume hood where the item is placed). Typically, room temperature is about 20°C to about 30°C, or about 22°C to about 27°C, or about 25°C.

[0224] The anti-solvent addition method (also known as anti-solvent crystallization, precipitation crystallization, salting out, or forced crystallization) is a method in which one or more anti-solvents are added to a solution in which the target product has been dissolved in a good solvent, resulting in a state in which the product is slightly dissolved in the solution, thereby making the solution supersaturated and then precipitating crystals.The anti-solvent reverse addition method is a method in which the target product is dissolved in a good solvent, and then the solution is added to one or more anti-solvents, resulting in a state in which the product is slightly dissolved in the solution, thereby making the solution supersaturated and then precipitating crystals.

[0225] A poor solvent has a lower ability to dissolve the target product than a good solvent, for example, by more than 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% less. The poor solvent in a system is relative. The good solvent and poor solvent may be polar or nonpolar, such as dimethylformamide (DMF), dimethyl sulfoxide (DMSO), water, alcohol solvents, ether solvents, ketone solvents, ester solvents, alkane solvents, aromatic solvents, and nitrile solvents. Alcohol solvents include, but are not limited to, methanol, ethanol, propanol, isopropanol, 1,3-propanediol, 1,2-propanediol, chlorobutanol, or combinations thereof. Ether solvents include, but are not limited to, tetrahydrofuran, methyl tert-butyl ether, 1,4-dioxane, or combinations thereof. Ketone solvents include, but are not limited to, acetone, methyl ethyl ketone, 4-methyl-2-pentanone, or combinations thereof. Ester solvents include, but are not limited to, ethyl acetate, isopropyl acetate, n-butyl acetate, tert-butyl acetate, or combinations thereof. Alkane solvents include, but are not limited to, methylene chloride, chloroform, n-hexane, cyclohexane, pentane, n-heptane, or combinations thereof. Aromatic solvents include, but are not limited to, benzene, toluene, or combinations thereof. Nitrile solvents include, but are not limited to, acetonitrile, malononitrile.

[0226] Antisolvent addition and antisolvent reverse addition can be performed by intermittent, semi-intermittent, or continuous crystallization. When adding an antisolvent to a solution (antisolvent crystallization) or a product solution to an antisolvent (antisolvent reverse crystallization), the antisolvent may be added dropwise at a constant rate, or the rate may be increased gradually after the initial dropwise addition.

[0227] The disclosures of various publications, patents and published patent specifications cited herein are hereby incorporated by reference in their entireties.

[0228] The technical solutions of the present invention will be described clearly and completely below with reference to the embodiments of the present invention, but it is clear that the described embodiments are only a part of the embodiments of the present invention, and are not all of them. Based on the embodiments of the present invention, all other embodiments that can be obtained by those skilled in the art without any creative efforts are all included in the protection scope of the present invention.

[0229] Example 1: Synthesis of Compound A2

[0230] [ka]

[0231] The experimental procedure is as follows.

[0232] Step 1:

[0233] [ka]

[0234] A 500 mL single-neck flask was charged with 1 (11.3 g, 23.7 mmol), bis(pinacolato)diboron (9.06 g, 35.6 mmol), bis(diphenylphosphino)ferrocenepalladium dichloride (1.74 g, 2.37 mmol), potassium acetate (6.99 g, 71.3 mmol), and dimethyl sulfoxide (150 mL). The mixture was protected with nitrogen and reacted at 95 °C for 16 h. The reaction was quenched with water (300 mL), extracted with ethyl acetate (150 mL x 3), washed with saturated brine (150 mL x 2), and spin-dried to give the desired product as a black solid (13.0 g, crude). LC-MS: 463 [M+Na] +

[0235] Step 2:

[0236] [ka]

[0237] A 500 mL three-neck flask was charged with 3 (15 g, 74.6 mmol), triethylamine (22.7 g, 223.8 mmol), and dichloromethane (150 mL). Methanesulfonyl chloride (12.1 g, 111.9 mmol) was added in an ice bath, and the mixture was allowed to react for 1 h. The mixture was quenched with water (300 mL), extracted with dichloromethane (100 mL x 3), washed with saturated brine, dried over anhydrous sodium sulfate, spin-dried, and purified by column chromatography (petroleum ether:ethyl acetate = 40:1) to give the desired product as a yellow solid (20.7 g, crude). LC-MS: 280 [M+H] +

[0238] Step 3:

[0239] [ka]

[0240] A 500 mL single-neck flask was charged with 5 (7.27 g, 49.4 mmol) and DMF (200 mL). 60% NaH (2.96 g, 74.1 mmol) was added in batches at 0 °C. The reaction was allowed to proceed at room temperature for 1 h. 4 (20.7 g, 74.1 mmol) was added, and the mixture was protected with nitrogen gas and reacted at 70 °C for 16 h. The mixture was quenched with water (500 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was washed with saturated brine (200 mL x 2), dried over anhydrous sodium sulfate, spin-dried, and separated by column chromatography (petroleum ether:ethyl acetate = 30:1) to obtain the desired product as a white solid (9.8 g, yield: 61.2%). LC-MS: 330 [M+H] +

[0241] Step 4:

[0242] [ka]

[0243] In a 500 mL single-neck 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 mixture was allowed to react at room temperature for 16 h. The mixture was concentrated at low temperature, diluted with DCM (200 mL), quenched with ice water, adjusted to pH 10 with aqueous ammonia at 0 °C, extracted with DCM (200 mL x 3), and the organic phase was washed with saturated brine (200 mL x 2), dried over anhydrous sodium sulfate, and spin-dried to give 6.59 g of the desired product as a white solid (yield: 96.6%). LC-MS: 230 [M+H] +

[0244] Step 5:

[0245] [ka]

[0246] In a 250 mL three-neck flask, 7 (5.53 g, 24.0 mmol) and TEA (10 mL, 72.1 mmol) were dissolved in THF (80 mL) and reacted at room temperature for 1 h. The mixture was cooled to 0 °C in an ice bath, and CD3I (1.65 mL, 26.4 mmol) was added dropwise. The mixture was protected with nitrogen gas and reacted at room temperature for 2 h. The mixture was quenched with water and extracted with DCM (100 mL x 3). The organic phase was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, and spin-dried to give the desired product (3.1 g, yellow oil) (yield: 52.2%). LC-MS: 248 [M+H] +

[0247] Step 6:

[0248] [ka]

[0249] A 500 mL single-neck flask was charged with 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 / HO (150 mL / 30 mL). The mixture was protected with nitrogen and reacted at 95 °C for 2.5 h. The reaction mixture was quenched with water (300 mL), extracted with ethyl acetate (150 mL x 3), washed with saturated brine (150 mL x 2), dried over anhydrous sodium sulfate, spin-dried, and flash purified to give the desired product as a black oil (3.3 g, crude). LC-MS: 563 [M+H] +

[0250] Step 7:

[0251] [ka]

[0252] A 250 mL three-neck flask was charged with 9 (3.3 g, 7.14 mmol), Fe (2.0 g, 35.7 mmol), ammonium chloride (1.93 g, 35.7 mmol), ethanol (40 mL), and water (8 mL), and the mixture was reacted at 85 °C for 2.5 h. The mixture was filtered under suction, and the filtrate was concentrated to give 3.5 g of the desired product as a crude black solid. LC-MS: 533 [M+H] +

[0253] Step 8:

[0254] [ka]

[0255] In a 250 mL three-neck 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 mixture was allowed to react at room temperature for 1 h. The mixture was concentrated at low temperature, diluted with DCM (100 mL), quenched with ice water, adjusted to pH 10 with aqueous ammonia at 0 °C, and extracted with DCM (100 mL x 3). The organic phase was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, spin-dried, and purified by preparative HPLC to obtain 450 mg of the desired product as a white solid (yield: 15.3%). LC-MS: 449 [M+H]+. 1 H 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).

[0256] Example 2: Synthesis of Compound B2

[0257] [ka]

[0258] The experimental procedure is as follows.

[0259] Step 1:

[0260] [ka]

[0261] A 2000 mL single-neck flask was charged with 1 (25.0 g, 102 mmol), an aqueous solution of chloroacetaldehyde (12.0 g, 154 mmol), and acetone (500 mL), and the mixture was reacted at 50 °C for 16 h. The mixture was spin-dried and separated by column chromatography (MeOH in DCM, 0% to 10%, v / v) to obtain the desired product as a yellow oil (9.20 g, yield: 53.4%). LC-MS: 169 [M+H] +

[0262] Step 2:

[0263] [ka]

[0264] A 1000 mL single-neck flask was charged with 3 (4.00 g, 23.8 mmol), triethylamine (6.01 g, 59.5 mmol), and tetrahydrofuran (200 mL). Deuterated iodomethane (3.62 g, 25.0 mmol) was added at room temperature and stirred for 2 hours. The mixture was quenched with water (250 mL), spin-dried, extracted with ethyl acetate (150 mL x 3), dried over anhydrous sodium sulfate, and spin-dried. Column chromatography (MeOH in DCM, 0% to 10%, v / v) afforded the desired product as a yellow solid (2.70 g, yield: 61.3%). LC-MS: 186 [M+H] +

[0265] Step 3:

[0266] [ka]

[0267] A 500 mL three-neck flask was charged with 4 (2.70 g, 14.6 mmol) and tetrahydrofuran (100 mL). Protected with nitrogen gas, n-butyllithium (2.4 M in tetrahydrofuran, 7.30 mL, 17.5 mmol) was added dropwise at -78 °C. The mixture was stirred for 1 h while maintaining the temperature. Next, n-tributyltin chloride (7.14 g, 21.9 mmol) was added dropwise and the reaction was continued at -78 °C for 1 h. After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride (100 mL) and extracted with ethyl acetate (120 mL x 3). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, suction filtered, and concentrated under reduced pressure to give a yellow oily residue containing no desired product. It could be used directly in the next step. (7.05 g, crude product). LC-MS: 476 [M+H] +

[0268] Step 4:

[0269] [ka]

[0270] A 3L single-neck flask was charged with 6 (59.4g, 707mmol), tetrahydrofuran (1.5L), DHP (68.4g, 813mmol), and PPTS (3.55g, 14.1mmol). The mixture was then reacted at room temperature for 16 hours. The mixture was quenched with saturated sodium bicarbonate (1000mL*3), washed with saturated brine (500mL), dried over anhydrous sodium sulfate, and spin-dried to give the desired product as a colorless oil (120g, crude). It could be used directly in the next step. LC-MS: 169 [M+H] +

[0271] Step 5:

[0272] [ka]

[0273] A 3 L single-neck flask was charged with 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). The mixture was protected with nitrogen and reacted at room temperature for 16 h. The reaction mixture was washed with saturated ammonium chloride (1000 mL) and saturated brine (500 mL), dried over anhydrous sodium sulfate, and spin-dried to give the desired compound as a yellow oil (59.4 g, crude product). It was suitable for direct use in the next step. LC-MS: 275 [M+H] +

[0274] Step 6:

[0275] [ka]

[0276] A 2000 mL three-neck flask was charged with 9 (50.3 g, 184 mmol), tetrahydrofuran (500 mL), 10 (40.1 g, 184 mmol), and triphenylphosphine (72.3 g, 276 mmol). The mixture was protected with nitrogen gas and stirred at room temperature. Diethyl azodicarboxylate (55.8 g, 276 mmol) was added. The mixture was allowed to react at room temperature for 16 hours. The mixture was then spin-dried and purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain the desired product as a yellow solid (70.2 g, yield: 80.6%). LC-MS: 476 [M+H] +

[0277] Step 7:

[0278] [ka]

[0279] A 1000 mL single-neck flask was charged with 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). The mixture was protected with nitrogen and stirred at 90 °C for 5 h. The mixture was diluted with ethyl acetate (400 mL), washed with saturated aqueous ammonium chloride (300 mL x 3), spun dry, and separated by column chromatography (MeOH in DCM, 0% to 5%, v / v) to give the desired product as a yellow solid (2.50 g, yield: 40.5%). LC-MS: 580 [M+H] +

[0280] Step 8:

[0281] [ka]

[0282] In a 500 mL single-neck flask, 12 (2.50 g, 4.32 mmol), reduced iron powder (1.21 g, 21.6 mmol), ammonium chloride (1.14 g, 21.6 mmol), ethanol (100 mL), and water (20 mL) were reacted at 80 °C for 2 h. The mixture was diluted with dichloromethane (100 mL), suction filtered, spin-dried, and separated by column chromatography (MeOH in DCM, 0% to 12%, v / v) to obtain the desired product as a yellow solid (1.40 mg, 59.1%). LC-MS: 550 [M+H] +

[0283] Step 9:

[0284] [ka]

[0285] A 250 mL single-neck flask was charged with 13 (1.02 g, 1.86 mmol) and tetrahydrofuran (25 mL). The mixture was stirred in an ice bath, and then a 4 M / L solution of hydrochloric acid gas in 1,4-dioxane (5 mL) was added dropwise. The mixture was allowed to react at room temperature for 20 minutes, then spin-dried, dissolved in dichloromethane, adjusted to pH 9 with aqueous ammonia, and spin-dried in dichloromethane (30 mL x 3). The desired product was obtained as a pale yellow solid (350 mg, 40.5%) by high-pressure liquid phase 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).

[0286] Example 3: Synthesis of Compound C

[0287] [ka]

[0288] The experimental procedure is as follows.

[0289] Step 1:

[0290] [ka]

[0291] A 2000 mL single-neck flask was charged with 1 (7.86 g, 30.0 mmol) and tetrahydrofuran (500 mL). Lithium aluminum deuteride hydride (3.15 g, 75.0 mmol) was added in batches at 0 °C, and the reaction was allowed to proceed at 0 °C for 1 hour. The mixture was quenched with acetic acid (50 mL), extracted with ethyl acetate (500 mL x 3), washed with saturated brine (500 mL x 3), and spin-dried to give the desired product as a yellow solid (3.50 g, crude). LC-MS: 259 [M+Na] +

[0292] Step 2:

[0293] [ka]

[0294] A 250 mL three-necked round-bottom flask was charged with 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). The flask was protected with nitrogen and heated at room temperature. 3 (2.50 g, 14.8 mmol) was added and the mixture was stirred for 16 h at room temperature. The mixture was diluted with dichloromethane (400 mL), washed with saturated aqueous ammonium chloride (300 mL x 3), spun dry, and purified by column chromatography (petroleum ether:ethyl acetate = 4:1) to give the desired product as a yellow solid (1.03 g, yield: 25.2%). LC-MS: 299 [M+Na] +

[0295] Step 3:

[0296] [ka]

[0297] A 250 mL three-neck flask was charged with 4 (1.03 g, 3.73 mmol), tetrahydrofuran (30 mL), 5 (854 mg, 3.92 mmol), and triphenylphosphine (1.47 g, 5.60 mmol). Under nitrogen gas protection, diethylpropyl azodicarboxylate (1.13 g, 5.60 mmol) was added dropwise at 0°C. Under nitrogen gas protection, the reaction was allowed to proceed at room temperature for 16 hours. The mixture was spin-dried and purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain the desired product as a yellow solid (1.30 g, yield: 73.2%). LC-MS: 499 [M+Na] +

[0298] Step 4:

[0299] [ka]

[0300] A 100 mL single-neck flask was charged with 6 (300 mg, 0.630 mmol), bis(pinacolato)diboron (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). The mixture was reacted at 90 °C for 16 h 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 spin-dried to give the desired product as a brown oil (400 mg, crude). LC-MS: 443 [M+H]+

[0301] Step 5:

[0302] [ka]

[0303] A 100 mL single-neck flask was charged with 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). The mixture was protected with nitrogen and reacted at 90 °C for 2 h. 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, spin-dried, and purified by column chromatography (dichloromethane:methanol = 18:1) to obtain the desired product as a yellow solid (250 mg, yield: 49.2%). LC-MS: 562 [M+H] +

[0304] Step 6:

[0305] [ka]

[0306] 9 (250 mg, 0.446 mmol), reduced iron powder (125 mg, 2.23 mmol), ammonium chloride (118 mg, 2.23 mmol), ethanol (10 mL), and water (2 mL) were added to a 100 mL single-neck flask and reacted at 80 °C for 2 h. The mixture was diluted with dichloromethane (50 mL), suction filtered, spin-dried, and separated by column chromatography (dichloromethane:methanol = 10:1) to obtain the desired product as a yellow solid (220 mg, 93.0%). LC-MS: 532 [M+H]+

[0307] Step 7:

[0308] [ka]

[0309] A 250 mL single-neck flask was charged with 10 (220 mg, 0.414 mmol) and tetrahydrofuran (10 mL). After stirring in an ice bath, 3 mL of 4 M / L hydrochloric acid in 1,4-dioxane was added dropwise. The mixture was allowed to react at room temperature for 20 minutes, then spin-dried, dissolved in dichloromethane, adjusted to pH 9 with aqueous ammonia, and spin-dried in dichloromethane (30 mL x 3). High-pressure liquid phase chromatography afforded the desired product as a pale yellow solid (85.0 mg, 45.9%). 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).

[0310] Example 4: Synthesis of Compound D

[0311] [ka]

[0312] The experimental procedure is as follows.

[0313] Step 1:

[0314] [ka]

[0315] A 100 mL three-neck flask was charged with 1 (3.0 g, 30.61 mmol) and tetrahydrofuran (30 mL). The mixture was protected with nitrogen gas. At -78 °C, n-butyllithium (2.4 M in tetrahydrofuran, 12.75 mL, 30.61 mmol) was added dropwise. The mixture was stirred for 1 hour while maintaining the temperature. 2 (2.0 g, 30.61 mmol) was then added dropwise. The mixture was then reacted at -78 °C for 1 hour, and then warmed to room temperature and reacted for 2 hours. After completion of the reaction, the mixture was quenched with saturated aqueous ammonium chloride (30 mL) and extracted with ethyl acetate (100 mL x 3). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, suction filtered, and concentrated under reduced pressure to give a yellow oily product, which did not contain the desired product. It could be used directly in the next step. (6.1 g, crude product). LC-MS: 163 [M+H] +

[0316] Step 2:

[0317] [ka]

[0318] A 250 mL single-neck flask was charged with 3 (6.1 g, 37.65 mmol), tetrahydrofuran (50 mL), DHP (4.76 g, 56.63 mmol), and PPTS (158.8 mg, 0.63 mmol). The mixture was then reacted at room temperature for 16 hours. The mixture was quenched with saturated sodium bicarbonate (100 mL x 3), washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and spin-dried to give the desired product as a colorless oil (8.5 g, crude). It could be used directly in the next step. LC-MS: 247 [M+H] +

[0319] Step 3:

[0320] [ka]

[0321] A 250 mL single-neck flask was charged with 4 (8.5 g, 34.55 mmol), methanol and dichloromethane (10 mL / 10 mL), and potassium carbonate (7.15 g, 51.83 mmol), and the mixture was allowed to react at room temperature for 3 hours. The filtrate was suction filtered, concentrated, and spin-dried to give the desired product as a colorless oil (6.3 g, crude). It could be used directly in the next step. LC-MS: 175 [M+H] +

[0322] Step 4:

[0323] [ka]

[0324] A 250 mL single-neck flask was charged with 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). The mixture was protected with nitrogen and reacted at room temperature for 16 h. The reaction mixture was washed with saturated ammonium chloride (200 mL x 3) and saturated brine (100 mL), dried over anhydrous sodium sulfate, spin-dried, and purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to give the target compound as a yellow oil (3.2 g). LC-MS: 281 [M+H] +

[0325] Step 5:

[0326] [ka]

[0327] A 100 mL three-neck flask was charged with 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). The mixture was protected with nitrogen gas and diisopropyl azodicarboxylate (1.624 g, 8.04 mmol) was added with stirring at room temperature. The mixture was allowed to react for 16 hours at room temperature, spin-dried, and purified by column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain the desired product as a yellow solid (450 mg, yield: 17.50%). LC-MS: 481 [M+H] +

[0328] Step 6:

[0329] [ka]

[0330] A 100 mL single-neck flask was charged with 9 (450 mg, 0.94 mmol), bis(pinacolato)diboron (359 mg, 1.41 mmol), bis(diphenylphosphino)ferrocenepalladium dichloride (35 mg, 0.047 mmol), potassium acetate (276 mg, 2.82 mmol), and dimethyl sulfoxide (10 mL). The mixture was protected with nitrogen and reacted at 90 °C for 16 h. The reaction was quenched with water (30 mL), extracted with ethyl acetate (50 mL x 3), washed with saturated brine (100 mL x 2), and spin-dried to give the desired product as a black solid (500 mg, crude). LC-MS: 447 [M+H] +

[0331] Step 7:

[0332] [ka]

[0333] A 100 mL single-neck flask was charged with 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-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (39 mg, 0.082 mmol), potassium phosphate (343 mg, 1.62 mmol), DMF (10 mL), and HO (2 mL), protected with nitrogen gas, and stirred at 95 °C for 1.5 h. The reaction mixture was quenched with water (30 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, spin-dried, and separated by TLC (dichloromethane:methanol = 15:1) to give the desired product as a brown liquid (266 mg, yield: 57.9%). LC-MS: 566 [M+H] +

[0334] Step 8:

[0335] [ka]

[0336] 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 to a 100 mL one-neck flask and reacted at 80 °C for 1 h. The mixture was cooled to room temperature, filtered under suction, and the filtrate was concentrated. Separation by TLC (methanol:dichloromethane = 1:10, v / v) afforded the desired product as a yellow solid (95 mg, 37.6%). LC-MS: 536 [M+H] +

[0337] Step 9:

[0338] [ka]

[0339] A 50 mL single-neck flask was charged with 13 (95 mg, 0.17 mmol) and tetrahydrofuran (5 mL). The mixture was stirred in an ice bath, and then a 4 M / L solution of hydrochloric acid in 1,4-dioxane (0.01 mL, 0.34 mmol) was added dropwise. The mixture was allowed to react at room temperature for 1 hour, and the mixture was spin-dried at 0°C. A black solid (15 mg, 18.7%) was obtained by high-pressure liquid phase chromatography. 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).

[0340] Example 5: Free base crystalline form A of compound A2 Compound A2 prepared in Example 1 was passed through a silica gel column to obtain 1.6 g of crude product (approximately 80% purity). This crude product was then separated by preparative HPLC, concentrated to 100 mL, adjusted to pH 9 with sodium bicarbonate solution, and extracted three times with dichloromethane (100 mL). The organic phase was washed once with saturated aqueous sodium chloride solution (80 mL), dried over sodium sulfate, and spin-dried to obtain 570 mg of a pale yellow solid (97% purity). This solid was slurried in 50 mL of petroleum ether / ethyl acetate (3:1) and filtered to obtain 460 mg of an off-white solid (purity >99%).

[0341] The following manufacturing process was repeated: (1) 230.9 mg of the above free base sample was weighed into a 20 mL glass bottle, and 1 mL of IPAc was added to obtain a clear solution; (2) After stirring (1000 rpm) at room temperature for about 5 minutes, a large amount of solid precipitated, and then 1 mL of IPAc was added; (3) After suspending and stirring at room temperature for about 1 day, the mixture was centrifuged (10,000 rpm, 2 minutes) to obtain a solid; (4) After vacuum drying at room temperature for 3 hours, a total of 195.4 mg of sample was collected (recovery rate: 84.6%).

[0342] The XRPD pattern of the sample is shown in Figure 1, and the data tables of its XRPD test parameters and results are shown in Tables 1 and 2, respectively, which indicate that the sample is crystalline, designated free-state crystalline form A, and repeatable. The entire list of peaks or corresponding d values ​​in Table 2, or a subset thereof, and an XRPD pattern substantially similar to that of Figure 1, may be sufficient to characterize the crystalline form.

[0343] The TGA / DSC pattern of the free base crystalline form A of Compound A2 is shown in Figure 2, and the test parameters of TGA and DSC are shown in Table 3. In Figure 2, the TGA results show that there is a 1.3% weight loss upon heating from room temperature to 170°C, and the DSC results show that there is one sharp endothermic peak at 168.8°C (onset temperature).

[0344] Free base crystalline form A of compound A2 1 The 1 H NMR spectrum is shown in Figure 3. 1 H NMR results indicate that the molar ratio of residual solvent IPAc to free base crystalline form A is 0.01:1 (corresponding to a TGA weight loss of 0.2%).

[0345] The HPLC chromatogram of the free base crystalline form A of Compound A2 is shown in FIG. 4, and the results are shown in Table 4.

[0346] The free base crystalline form A of Compound A2 is presumed to be an anhydrous crystalline form.

[0347] [Table 1]

[0348] [Table 2]

[0349] [Table 3]

[0350] [Table 4]

[0351] Solid-state stability of free base crystalline form A

[0352] The XRPD patterns of the initial sample of the free base crystalline form A and the sample after standing in a sealed space at 60° C. for one day are shown in FIG. 5, and the HPLC results are shown in Table 5.

[0353] [Table 5]

[0354] Example 6: Free base crystalline form B of compound A2 (1) Weigh 9.9g of free base sample into a 20mL glass bottle (e.g., as described in Example 5), add 2mL of EtOAc to dissolve the sample, and filter through a 0.45μm PTFE filter membrane to obtain a clear API solution; (2) Add the antisolvent toluene dropwise to the API solution, stirring magnetically (~1000 rpm) while adding the solution dropwise, until a total of 10 mL of toluene is added; (3) The resulting clear solution was stirred at room temperature for up to 2 hours, then transferred to 5°C and stirred for approximately 15 hours, still resulting in a clear solution; (4) After being transferred to -20°C and stirred for approximately 6 hours, the solution remained clear; (5) The clear solution was transferred to room temperature and evaporated for 8 days to give a solid.

[0355] The XRPD pattern of the sample is shown in Figure 6, its XRPD test parameters are shown above in Table 1, and the resulting data table is shown in Table 6, which showed it to be crystalline and designated free-state crystalline form B. The entire list of peaks or corresponding d values ​​in Table 6, or a subset thereof, and an XRPD pattern substantially similar to that of Figure 5 may be sufficient to characterize the crystalline form.

[0356] [Table 6]

[0357] The TGA / DSC pattern of the free base crystalline form B of Compound A2 is shown in Figure 7, and the test parameters for TGA and DSC are shown in Table 3 above. The TGA results show that the sample loses 13.2% weight when heated from room temperature to 70°C, and loses 8.5% weight when subsequently heated to 170°C. The DSC results show that the sample has four endothermic peaks at 59.5°C, 95.6°C, 150.8°C, and 160.9°C (peak temperatures).

[0358] Free base crystalline form B of compound A2 1 The 1 H NMR spectrum is shown in Figure 8. 1 H NMR results indicated that the molar ratio of residual solvent EtOAc to free base was 0.8:1 (corresponding to a TGA weight loss of 18.1%).

[0359] The XRPD patterns of the free base crystalline form B of Compound A2 before and after standing at room temperature are shown in Figure 9. The results show that after standing at room temperature in a closed space for 5 days, the diffraction peaks of the free base crystalline form A appeared, indicating that the free base crystalline form B tends to convert to the free base crystalline form A when standing at room temperature.

[0360] The XRPD patterns of the free base crystalline form B of Compound A2 before and after nitrogen purging are shown in Figure 10. The results showed that the free base crystalline form B (containing one diffraction peak of crystalline form A) changed its crystalline form after purging with nitrogen for 20 minutes at 30°C. This is presumably because the free base crystalline form B changed its crystalline form due to the removal of EtOAc after nitrogen purging.

[0361] Taking the specific results together, it is assumed that the free base crystalline form B is an EtOAc solvate.

[0362] Example 7: Preparation of the hydrochloride salt of compound A2: 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 1.11 mL of a 1N solution of hydrochloric acid in ethyl ether was added dropwise. After the dropwise addition was completed, the mixture was stirred at room temperature for 30 minutes. TLC showed that all the starting materials had disappeared. The mixture was purged with nitrogen gas for 10 minutes and then concentrated under reduced pressure at 10°C to obtain a white solid (purity >99%). This solid was placed in a freeze dryer for 12 hours to obtain 108 mg of the hydrochloride salt of A2 (white, purity >99%).

[0363] The XRPD pattern of the hydrochloride salt of Compound A2 is shown in Figure 11, the XRPD test parameters are shown in Table 1 of Example 5, and the data table of its XRPD results is shown in Table 7, which showed it to be crystalline and designated hydrochloride crystalline form A. The entire list of peaks or corresponding d values ​​in Table 7, or a subset thereof, and an XRPD pattern substantially similar to that of Figure 11 may be sufficient to characterize the crystalline form.

[0364] The TGA / DSC pattern of the hydrochloride salt of Compound A2 is shown in Figure 12, and the test parameters for TGA and DSC are shown in Table 3 of Example 5. The TGA results show that there is an 8.6% weight loss when heated from room temperature to 150°C. The DSC results show that there are two endothermic peaks at 81.9°C and 156.0°C (peak temperatures).

[0365] [Table 7]

[0366] The XRPD patterns of the free base crystalline form A of Compound A2 and the hydrochloride salt of Compound A2 are overlaid and shown in Figure 13. The XRPD results for both are essentially the same, but the hydrochloride salt has a lower degree of crystallinity.

[0367] Example 8 Procedure for identification of microsomal metabolites: The compounds (test compounds and numbers are listed below; compounds A1 and B1 were prepared with reference to the related methods disclosed in CN110396087A) were weighed and dissolved in dimethyl sulfoxide to a 20 mM solution. The compound stock solution was diluted with 50% acetonitrile (v / v) to a concentration of 1.0 mM to prepare the working solution. Liver microsomes (20 mg / mL) were diluted to 1.27 mg / mL with 50 mM dipotassium hydrogen phosphate buffer to prepare the liver microsome working solution. Reduced coenzyme was weighed and added to 3307 μL of phosphate buffer (50 mM) to prepare a 5.0 mM solution to prepare the reduced coenzyme working solution. For the T=60 min sample, 4 μL of sample working solution (1.0 mM) was added, followed by 316 μL of liver microsome working solution (1.27 mg / mL), and finally 80 μL of reduced coenzyme working solution was added to initiate the reaction. To the T=0 min sample, 316 μL of liver microsome working solution (1.27 mg / mL) was added, followed by 80 μL of reduced coenzyme working solution to initiate the reaction; the sample was not incubated. After 60 min of incubation at 37°C, the enzymatic reaction was terminated by adding 1200 μL of stop solution. To the T=0 min sample, 4 μL of sample working solution (1.0 mM) was added. The sample plate was placed on a vortexer and vortexed at 600 rpm for 5 min. The mixture was then centrifuged at 4000 rpm for 10 min. The supernatant was removed, mixed, and dried by blowing with nitrogen gas at room temperature. The residue dried by blowing with nitrogen gas was redissolved in 300 μL of 10% acetonitrile (0.1% FA) solvent and centrifuged at 4000 rpm for 15 min. The supernatant was transferred to a detection plate and prepared for mass spectrometry analysis. Mass spectrometry analysis was performed using an LC / Q-Exactive Plus.

[0368] 7-Ethoxycoumarin (10 μM) was chosen as a positive control compound and run in parallel with that compound.

[0369] [ka]

[0370] The remaining rate of parent nuclei after 60 minutes is shown in the table below.

[0371] [Table 8]

[0372] Example 9 Pharmacokinetics Experiments Pharmacokinetics experiments using rats Six male Sprague Dawley rats were administered a single dose by intravenous injection (iv, n=3) and oral administration (po, n=3) in sequence. For intravenous injection, the compound was dissolved in 10% DMSO / 30% PEG400 / 60% water at 0.25 mg / mL or 0.5 mg / mL and administered at a volume of 2 mL / kg. For oral administration, the compound was suspended in 0.5% methylcellulose at 0.6 mg / mL or 2.0 mg / mL and administered at a volume of 5 mL / kg. The specific dosages are shown in the table below.

[0373] [Table 9]

[0374] After administration, blood samples were taken for intravenous injection and oral administration at 0.0833, 0.25, 0.5, 1, 2, 4, 8, and 24 hours, and the concentrations of the compounds in the samples were measured by LC-MS / MS with a lower limit of quantification of 1 ng / mL. The pharmacokinetic parameters of the compounds were calculated using the WinNolin nonventricular model.

[0375] The structures and numbers of the compounds involved in this example are shown below.

[0376] [ka]

[0377] result After intravenous administration, the internal exposure (AUC last) were 116hr*ng / mL, 247hr*ng / mL, 74.8hr*ng / mL, and 28.7hr*ng / mL, respectively, and the mean total clearance (CL) was 129mL / min / kg, 71mL / min / kg, 101mL / min / kg, and 151mL / min / kg, respectively.

[0378] After oral administration, the internal exposure of A1, A2, C, and D (AUC last ) were 79.8ng*hr / mL, 572ng*hr / mL, 42.8ng*hr / mL, and 9.94ng*hr / mL, respectively. Compared with the intravenous data, the oral bioavailability in rats was 6.84%, 23.2%, 9.52%, and 5.2%, respectively.

[0379] Pharmacokinetics study using cynomolgus monkeys Three male cynomolgus monkeys were given a single intravenous injection (1 mg / kg, n=3) at a single dose, allowed to leach for one week, and then orally administered (10 mg / kg, n=3). For intravenous injections, the compound was dissolved in water or 10% DMSO / 30% PEG400 / Water at a concentration of 1.0 mg / mL and administered at a volume of 1 mL / kg. For oral administration, the compound was dissolved in water or 0.5% methylcellulose / 1.7 meq 1N hydrochloric acid at a concentration of 2.0 mg / mL and suspended in water at a volume of 5 mL / kg.

[0380] After administration, blood samples were taken for intravenous injection and oral administration at 0.0833, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours, and the concentrations of the compounds in the samples were measured by LC-MS / MS with a lower limit of quantification of 1 ng / mL. The pharmacokinetic parameters of the compounds were calculated using the WinNolin nonventricular model.

[0381] result After intravenous administration, the internal exposure of A1 and A2 (AUC last ) were 881 hr*ng / mL and 949 hr*ng / mL, respectively, and the mean total clearance (CL) was 18.9 mL / min / kg and 16.7 mL / min / kg, respectively.

[0382] After oral administration, the internal exposure of A1 and A2 (AUC last ) were 388 ng*hr / mL and 1758 ng*hr / mL, respectively. Compared with the intravenous data, oral bioavailability in cynomolgus monkeys was 4.61% and 18.8%, respectively.

[0383] The above-mentioned are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, etc. made within the concept and principle of the present invention shall be included in the protection scope of the present invention.

[0384] The above embodiments and methods described in the present invention may vary based on the ability, experience, and preferences of those skilled in the art.

[0385] In the present invention, the steps of the method are merely described in a certain order, but the order of the steps of the method is not limited thereto.

Claims

1. A compound represented by general formula II, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, 1. A compound, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, characterized in that said compound has the structure: 【Chemical 1】 However, the E ring is 【Chemistry 2】 Selected from: In the E ring, each R 0 are independently -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; provided that H attached to the C atom or heteroatom may be replaced by deuterium; R 1 teeth, 【Chemistry 3】 、-CH 3 、 【Chemistry 4】 、-CH 2 D、-CHD 2 、-CD 3 、 【Chemistry 5】 Selected from: R 2 Ha-NO 2 , -N(C 0 ~ 10 alkyl) (C 0 ~ 10 alkyl), —OC 0 ~ 10 Alkyl and -OCF 3 wherein H bonded to a C atom or N atom may be replaced by deuterium; R 3 is -H, -D, halogen, -OC 0 ~ 10 Alkyl, —CO(C 0 ~ 10 alkyl), C 1 ~ 10 Selected from straight chain / branched alkyl, with the proviso that the H atom attached to the C atom may be replaced with deuterium; R 4 is -C≡C-R 10 and R 5 , R 6 , R 7 are independently —H, —D, halogen, —CN, C 1 ~ 3 Straight chain / branched alkyl, —OC 0 ~ 3 Alkyl, —CO(C 0 ~ 3 alkyl), N-containing C 1 ~ 3 straight chain / branched alkyl, or R 6 , R 7 , and R 6 and R 7 The carbon atom between 3 ~ 8 Cycloalkyl or —O— containing C 3 ~ 8 forming a heterocycloalkyl, wherein the H atom bonded to the C atom or the N atom may be replaced by deuterium; R 8 and R 9 are independently -H, -D, C 1 ~ 3 Selected from straight chain / branched alkyl, with the proviso that the H atom attached to the C atom may be replaced by deuterium; R 10 is H, -D, C 1 ~ 5 Straight / branched alkyl, C 3 ~ 10 cycloalkyl, 【Chemistry 6】 wherein one or more H atoms to which the C atom is attached may be replaced by deuterium; R 11 , R 12 are independently -H, -D, -CF 3 , -CHF 2 , -CH 2 F, 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, an aromatic five-membered ring group, or an aromatic six-membered ring group; or R 11 , R 12 , and R 11 and R 12 The carbon atom between 3 ~ 8 Cycloalkyl or C containing —O—, —S— 3 ~ 8 Heterocycloalkyl, C 4 ~ 9 Fused cycloalkyl, C 5 ~ 10 Spirocycloalkyl, C 4 ~ 9 Bridged cycloalkyl, C 3 ~ 7 Cyclolactam, C 3 ~ 7 Cyclolactone, C 3 ~ 7 forming a cycloketone, wherein one or more H atoms to which a C atom or heteroatom is attached may be replaced by deuterium; And the R 1 , R 4 , R 8 , R 9 At least one of the groups contains a deuterium atom.

2. R 11 and R 12 are independently -H, -D, -CF 3 , -CHF 2 , -CH 2 F, -CH 3 , -CH 2 CH 3 , -CH=CH 2 , 【Chemistry 7】 or R 11 , R 12 and R 11 and R 12 The carbon atom in between is 【Chemistry 8】 2. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein H bonded to a C atom or an N atom may be substituted with deuterium.

3. R 11 and R 12 are independently -H, -D, -CF 3 , -CHF 2 , -CDF 2 , -CH 2 F, -CD 2 F, -CH 3 , -CH 2 D, -CHD 2 , -CD 3 , -CH 2 CH 3 , -CH 2 CD 3 , 【Chemistry 9】 2. The compound of claim 1, wherein the compound is selected from:

4. Each R 0 are independently —H, —D, —CH 3 , -CH 2 CH 3 and -NH 2 2. The compound of claim 1, wherein the compound is selected from:

5. R 2 is -NH 2 , -NHD, -ND 2 and -NO 2 2. The compound of claim 1, wherein the compound is selected from:

6. R 3 is -H, -D, -F, -OCH 3 , -OCH 2 D, -OCHD 2 , -OCD 3 2. The compound of claim 1, wherein the compound is selected from:

7. R 5 , R 6 , R 7 are independently —H, —D, —F, —Cl, —CH 3 , -CH 2 D, -CHD 2 , -CD 3 , -OCH 3 , -COCH 3 , -CH 2 NH 2 , -CH 2 N (CH 3 ) 2 , -CN, -OCH 2 D, -OCHD 2 , -OCD 3 , -COCD 3 , -CH 2 N (CD 3 ) 2 , -CH 2 N (CH 3 ) (CD 3 2. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound is selected from the group consisting of:

8. R 8 and R 9 are independently —H, —D, —CH 3 , -CH 2 D, -CHD 2 , -CD 3 2. The compound of claim 1, wherein the compound is selected from:

9. 2. The compound of claim 1, wherein the compound is selected from the following structures: 【Chemistry 10-1】 【Chemistry 10-2】 【Chemistry 10-3】 【Chemistry 10-4】 【Chemistry 10-5】 【Chemistry 10-6】 【10-7】 【10-8】 【Chemistry 10-9】 【10-10】 【Chemistry 10-11】 【10-12】 【Chemistry 10-13】 【10-14】 【10-15】 【10-16】 【10-17】 【10-18】 【10-19】 【10-20】 【10-21】

10. 2. The compound of claim 1, wherein the compound is selected from the following structures: 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】

11. A crystalline form of the compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, The crystal is a crystal of 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol, and its XRPD pattern has characteristic peaks at at least three positions with 2θ values ​​of 13.1°±0.2°, 16.3°±0.2°, 17.5°±0.2°, and 23.8°±0.2°.

12. The crystal according to claim 11, further characterized in that the XRPD pattern of the crystal has characteristic peaks at at least three 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°.

13. The crystal according to claim 11, characterized in that the crystal has an XRPD pattern as shown in Figure 1-1 below. 【Table 1】

14. A crystalline form of the compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, The crystal is a crystal of 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol, and its XRPD pattern has characteristic peaks at at least three positions with 2θ values ​​of 5.7°±0.2°, 11.3°±0.2°, 22.7°±0.2°, and 23.5°±0.2°.

15. The crystal according to claim 14, further characterized in that the XRPD pattern of the crystal has characteristic peaks at at least three 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°.

16. The crystal according to claim 14, characterized in that the crystal has an XRPD pattern as shown in Figure 6 below. 【Table 2】

17. A crystalline form of the compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, The crystal is a crystal of 4-(3-(((2-amino-5-(1-(1-trideuteromethylpiperidin-4-yl)-1H-pyrazol-4-yl)pyridin-3-yl)oxy)methyl)phenyl)-2-methylbut-3-yn-2-ol hydrochloride, and the XRPD pattern thereof has characteristic peaks at at least three 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°.

18. The crystal of claim 17, wherein the XRPD pattern of the crystal further comprises characteristic peaks at at least three 2θ values ​​of 8.1°±0.2°, 12.1°±0.2°, 15.3°±0.2°, 18.0°±0.2°, and 21.4°±0.2°.

19. The crystal according to claim 17, characterized in that the crystal has an XRPD pattern as shown in Figure 11 below. 【Table 3】

20. A compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or a crystal according to any one of claims 11 to 19, and A pharmaceutical composition comprising a pharmaceutically acceptable excipient.

21. Use of the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the crystal according to any one of claims 11 to 19, or the pharmaceutical composition according to claim 20, in the manufacture of a medicament for the prevention and / or treatment of a tumor.

22. The tumor is selected from the group consisting of lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuroendocrine tumor, carcinoid tumor, gastrinoma, pancreatic islet cell tumor, mesothelioma, neurilemmoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or malignant lymphoma, squamous cell carcinoma, epithelial squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, 22. The use according to claim 21, characterized in that the cancer is selected from peritoneal cancer, 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 cancer, head and neck cancer, and malignant hematological tumors.

23. Use of the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the crystal according to any one of claims 11 to 19, or the pharmaceutical composition according to claim 20, in the manufacture of a medicament for the prevention and / or treatment of a disease caused by or associated with pathogen infection.

24. 24. The use according to claim 23, wherein the pathogen is a virus.

25. 25. The use according to claim 24, characterized in that the virus is selected from HBV, HIV, HCV, HPV, Ebola virus, Marburg virus, influenza virus, parainfluenza virus, dengue virus, SARS-CoV, SARS-CoV-2.

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