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WO2024169781A1 - Pyrimidinone derivative and pharmaceutical application thereof - Google Patents

Pyrimidinone derivative and pharmaceutical application thereof Download PDF

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Publication number
WO2024169781A1
WO2024169781A1 PCT/CN2024/076273 CN2024076273W WO2024169781A1 WO 2024169781 A1 WO2024169781 A1 WO 2024169781A1 CN 2024076273 W CN2024076273 W CN 2024076273W WO 2024169781 A1 WO2024169781 A1 WO 2024169781A1
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Prior art keywords
compound
pharmaceutically acceptable
added
stereoisomer
alkyl
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French (fr)
Chinese (zh)
Inventor
杨亚讯
赵杨杨
罗云富
陈曙辉
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Medshine Discovery Inc
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Medshine Discovery Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
    • C07D239/545—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

  • the present invention relates to a class of pyrimidone derivatives and applications thereof, and in particular to a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof.
  • Purinergic receptors are ion channel receptors with adenosine triphosphate (ATP) as ligand.
  • ATP adenosine triphosphate
  • purinergic receptor subtypes including 7 ion channel receptors (ionotropic receptors P2X), 9 metabotropic receptors (metabotropic receptors P2Y) and 4 adenosine receptors.
  • P2X3 receptor purinergic ligandgated ion channel 3 receptor
  • P2X3 receptor has unique and important physiological functions. P2X3 is selectively expressed in primary sensory neurons, mediating signal transduction and regulating a variety of physiological functions.
  • ATP can activate P2X3 receptors after binding to them, increasing the permeability of the cell membrane to Na + , K + , and Ca 2+ , especially the permeability change of Ca 2+ is the most obvious.
  • a large amount of ATP can be released, activating the presynaptic P2X3 receptor, causing a large amount of Ca 2+ influx, and the increase in intracellular calcium concentration activates protein kinase A (PKA) and protein kinase C (PKC), causing PKA and PKC phosphorylation, while promoting the release of glutamate, further activating N-methyl-D-aspartic acid receptor (NMDA), leading to the generation of excitatory postsynaptic currents, causing central sensitization.
  • PKA protein kinase A
  • PKC protein kinase C
  • NMDA N-methyl-D-aspartic acid receptor
  • P2X3 receptor expression can lead to the formation of pain sensitivity, participate in the signal transmission of pain, and P2X3 receptors can mediate the generation of inflammatory pain, neuropathic pain, cancer pain, etc.
  • P2X3 inhibitors can mediate the generation of inflammatory pain, neuropathic pain, cancer pain, etc.
  • the public in vivo research of P2X3 inhibitors is used to treat diseases such as cough and pain.
  • the discovery and application of new P2X3 inhibitors have broad prospects and are of great significance for the treatment of diseases such as cough and pain.
  • the present invention provides a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,
  • R 1 is selected from H, D, halogen, CN, C 1-3 alkyl or C 1-3 haloalkyl;
  • each R 2 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , -CH 2 OH, cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy, wherein the cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 Ra ;
  • each R 3 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R b ;
  • each R 4 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R c ;
  • R 5 and R 6 are each independently selected from H, D, F, CN and C 1-4 alkyl;
  • R 5 and R 6 and the carbon atom to which they are connected form a cyclopropyl group
  • R 7 and R 8 are each independently selected from H, D, F, OH, CN and C 1-4 alkyl;
  • R7 and R8 and the carbon atoms to which they are connected form a cyclopropyl group, a cyclobutyl group or an oxetanyl group;
  • R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and The C 1-4 alkoxy and C 1-4 alkylamino groups are each independently optionally substituted by 1, 2 or 3 substituents selected from F, Cl, Br, I and OH;
  • Each Ra is independently selected from D, F, Cl, Br, I, CN, NH2 and OH;
  • Each R b is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;
  • Each R c is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;
  • n is selected from 0, 1, 2, 3 and 4;
  • n is selected from 0, 1, 2, 3 and 4;
  • r is selected from 0, 1, 2, 3 and 4;
  • t is selected from 1, 2 and 3;
  • Ring A1 is selected from phenyl and 5-10 membered heteroaryl
  • Ring A2 is selected from phenyl and 5-6 membered heteroaryl
  • Ring A3 is selected from phenyl and 5-6 membered heteroaryl
  • E 1 is absent or selected from NR 10 ;
  • R 10 is selected from H and C 1-4 alkyl
  • R7 and R8 are not all H.
  • the present invention provides a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,
  • R 1 is selected from H, D, halogen, CN, C 1-3 alkyl or C 1-3 haloalkyl;
  • each R 2 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , -CH 2 OH, cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy, wherein the cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 Ra ;
  • each R 3 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R b ;
  • Each R 4 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy,
  • the C 1-3 alkyl and C 1-3 alkoxy groups are optionally substituted by 1, 2 or 3 R c ;
  • R 5 and R 6 are each independently selected from H, D, F, CN and C 1-4 alkyl;
  • R 5 and R 6 and the carbon atom to which they are connected form a cyclopropyl group
  • R 7 and R 8 are each independently selected from H, D, F, OH, CN and C 1-4 alkyl;
  • R7 and R8 and the carbon atoms to which they are connected form a cyclopropyl group, a cyclobutyl group or an oxetanyl group;
  • R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and The C 1-4 alkoxy and C 1-4 alkylamino groups are each independently optionally substituted by 1, 2 or 3 substituents selected from F, Cl, Br, I and OH;
  • Each Ra is independently selected from D, F, Cl, Br, I, CN, NH2 and OH;
  • Each R b is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;
  • Each R c is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;
  • n is selected from 0, 1, 2, 3 and 4;
  • n is selected from 0, 1, 2, 3 and 4;
  • r is selected from 0, 1, 2, 3 and 4;
  • t is selected from 1, 2 and 3;
  • Ring A1 is selected from phenyl and 5-6 membered heteroaryl
  • Ring A2 is selected from phenyl and 5-6 membered heteroaryl
  • Ring A3 is selected from phenyl and 5-6 membered heteroaryl
  • E 1 is absent or selected from NR 10 ;
  • R 10 is selected from H and C 1-4 alkyl
  • R7 and R8 are not all H.
  • the present invention provides a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,
  • R 1 is selected from H, D, halogen, CN, C 1-3 alkyl or C 1-3 haloalkyl;
  • each R 2 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , -CH 2 OH, cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy, wherein the cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 Ra ;
  • each R 3 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R b ;
  • each R 4 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R c ;
  • R 5 and R 6 are each independently selected from H, D, F, CN and methyl;
  • R 5 and R 6 and the carbon atom to which they are connected form a cyclopropyl group
  • R 7 and R 8 are each independently selected from H, D, F, OH, CN and methyl;
  • R7 and R8 and the carbon atoms to which they are connected form a cyclopropyl group, a cyclobutyl group or an oxetanyl group;
  • R 9 is selected from OH, C 1-4 alkoxy and C 1-4 alkylamino, wherein the C 1-4 alkoxy and C 1-4 alkylamino are each independently optionally substituted by 1, 2 or 3 substituents selected from F and OH;
  • Each Ra is independently selected from D, F, Cl, Br, I, CN, NH2 and OH;
  • Each R b is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;
  • Each R c is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;
  • n is selected from 0, 1, 2, 3 and 4;
  • n is selected from 0, 1, 2, 3 and 4;
  • r is selected from 0, 1, 2, 3 and 4;
  • t is selected from 1, 2 and 3;
  • Ring A1 is selected from phenyl and 5-6 membered heteroaryl
  • Ring A2 is selected from phenyl and 5-6 membered heteroaryl
  • Ring A3 is selected from phenyl and 5-6 membered heteroaryl
  • E 1 is absent or selected from NR 10 ;
  • R 10 is selected from H and C 1-4 alkyl.
  • R 1 is selected from H, D, F, Cl and methyl, and other variables are as defined in the present invention.
  • each R 2 mentioned above is independently selected from H, D, F, Cl, CN, methyl and methoxy, and other variables are as defined in the present invention.
  • each R 4 mentioned above is independently selected from H, D, F, Cl and methyl, and other variables are as defined in the present invention.
  • R 5 and R 6 are each independently selected from H, D, F and methyl, and other variables are as defined in the present invention.
  • the carbon atoms connected to R7 and R8 may be the same atom or different atoms.
  • the above Selected from -CH 2 -, -(CH 2 ) 2 -, -CH(CH 3 )-, -C(CH 3 ) 2 -, -(CH 2 ) 3 -, -CH2CH ( CH3 )-, -CH2C ( CH3 ) 2- and -CH2CH (F)-, and other variables are as defined herein.
  • the above is selected from -CH 2 -, -(CH 2 ) 2 -, -CH(CH 3 )-, -(CH 2 ) 3 -, -CH2CH ( CH3 )-, -CH2C ( CH3 ) 2- and -CH2CH (F)-, and other variables are as defined herein.
  • the above Selected from -CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )- and -CH 2 C(CH 3 ) 2 -, and other variables are as defined herein.
  • the above is selected from -CH 2 - and -CH 2 CH(CH 3 )-, and the other variables are as defined herein.
  • the above is selected from -CH 2 CH(CH 3 )-, and the other variables are as defined herein.
  • the above R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and
  • the C 1-4 alkoxy and C 1-4 alkylamino are each independently optionally substituted by 1, 2 or 3 F, and other variables are as defined herein.
  • the above R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and
  • the C 1-4 alkoxy and C 1-4 alkylamino are each independently optionally substituted by 1, 2 or 3 F, and other variables are as defined herein.
  • the above R 9 is selected from OH, methoxy, methylamino, ethylamino and
  • the methoxy group, methylamino group and ethylamino group are each independently optionally substituted with 1, 2 or 3 F groups, and other variables are as defined in the present invention.
  • the above R 9 is selected from OH, OCH 3 , NHS(O) 2 CH 3 , NHCH 3 , NHCH 2 CHF 2 and NHCH 2 CF 3 , and other variables are as defined in the present invention.
  • R 9 is selected from OH, and other variables are as defined in the present invention.
  • the ring A1 is selected from phenyl, Other variables are as defined in the present invention.
  • the ring A1 is selected from phenyl, Other variables are as defined in the present invention.
  • the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.
  • the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.
  • the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.
  • the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.
  • the above structural unit Selected from Other variables are as defined in the present invention.
  • the above structural unit Selected from Other variables are as defined in the present invention.
  • the above structural unit Selected from Other variables are as defined in the present invention.
  • the above structural unit Selected from Other variables are as defined in the present invention.
  • the above t is selected from 1, and other variables are as defined in the present invention.
  • the above t is selected from 2, and other variables are as defined in the present invention.
  • the above t is selected from 3, and other variables are as defined in the present invention.
  • the above r is selected from 0, 1 and 2, and other variables are as defined in the present invention.
  • the above r is selected from 1, and other variables are as defined in the present invention.
  • the above-mentioned ring A2 is selected from phenyl, and other variables are as defined in the present invention.
  • the above-mentioned ring A3 is selected from phenyl, and other variables are as defined in the present invention.
  • the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt is selected from the structure shown in formula (VII-1) and (III-1):
  • T1 is selected from CH and N; R1 , R2 , R3 , R4 , R5 , R6 , R7 , R8 , R9 , m, n, r and t are as defined in the present invention.
  • the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt is selected from the structure shown in formula (I-1) and (III-1):
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , m, n, r and t are as defined in the present invention.
  • the above-mentioned compound, its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compounds represented by formula (P-1) and (P-2):
  • T1 is selected from CH and N;
  • E 2 is selected from NH and CH 2 ;
  • R1 , R2 , R3 , R4 , R5 , R6 , R7 , R8 , R9 , m, n, r and t are as defined herein.
  • the present invention also provides the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:
  • the present invention also provides the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:
  • the present invention also provides the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:
  • the present invention also provides a crystalline form A of compound TM03, characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ angles: 13.69 ⁇ 0.20°, 15.32 ⁇ 0.20°, 17.30 ⁇ 0.20°, 19.41 ⁇ 0.20° and 21.45 ⁇ 0.20°;
  • the above-mentioned crystal form A is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ angles: 9.44 ⁇ 0.20°, 13.69 ⁇ 0.20°, 15.32 ⁇ 0.20°, 17.30 ⁇ 0.20°, 17.85 ⁇ 0.20°, 19.41 ⁇ 0.20°, 21.45 ⁇ 0.20° and 21.73 ⁇ 0.20°.
  • the above-mentioned crystal form A is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ angles: 7.57 ⁇ 0.20°, 9.44 ⁇ 0.20°, 13.69 ⁇ 0.20°, 15.32 ⁇ 0.20°, 17.30 ⁇ 0.20°, 17.85 ⁇ 0.20°, 18.55 ⁇ 0.20°, 19.41 ⁇ 0.20°, 20.24 ⁇ 0.20°, 20.44 ⁇ 0.20°, 21.45 ⁇ 0.20°, 21.73 ⁇ 0.20° and 23.48 ⁇ 0.20°.
  • the above-mentioned crystal form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2 ⁇ angles: 7.57 ⁇ 0.20°, 8.441 ⁇ 0.20°, 9.44 ⁇ 0.20°, 10.704 ⁇ 0.20°, 12.12 ⁇ 0.20°, 13.091 ⁇ 0.20°, 13.687 ⁇ 0.20°, 15.323 ⁇ 0.20°, 16.14 ⁇ 0.20°, 16.977 ⁇ 0.20°, 17.304 ⁇ 0.20°, 17.847 ⁇ 0.20°, 18.55 ⁇ 0.20°, 1 9.412 ⁇ 0.20°, 20.24 ⁇ 0.20°, 20.439 ⁇ 0.20°, 21.449 ⁇ 0.20°, 21.73 ⁇ 0.20°, 22.833 ⁇ 0.20°, 23.482 ⁇ 0.20°, 24.648 ⁇ 0.20°, 25.297 ⁇ 0.20°, 25.594 ⁇ 0.20°, 26.43 ⁇ 0.20°, 27.141 ⁇ 0.20°, 27.498 ⁇ 0.20°, 27.932 ⁇
  • the XRPD spectrum analysis data of the above-mentioned Form A is basically as shown in Table 1.
  • the XRPD spectrum of Form A of compound TM03 is substantially as shown in FIG6 .
  • the differential scanning calorimetry curve of the above-mentioned crystal form A has a starting point of an endothermic peak at 111.1°C ⁇ 5°C; and a starting point of an exothermic peak at 176.3°C ⁇ 5°C.
  • the DSC spectrum of the above-mentioned Form A is substantially as shown in FIG. 7 .
  • thermogravimetric analysis curve of the above-mentioned crystal form A shows a weight loss of 9.325% at 150°C.
  • the TGA spectrum of the above-mentioned crystal form A is basically as shown in Figure 8.
  • the present invention also provides a method for preparing the A crystal form of compound TM03, which comprises the following steps (a) to (c):
  • the present invention also provides a crystal form B of compound TM03, characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ angles: 7.06 ⁇ 0.20°, 14.23 ⁇ 0.20°, 19.33 ⁇ 0.20° and 20.91 ⁇ 0.20°;
  • the above-mentioned B crystal form has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2 ⁇ angles: 7.06 ⁇ 0.20°, 14.23 ⁇ 0.20°, 19.33 ⁇ 0.20°, 20.91 ⁇ 0.20° and 21.46 ⁇ 0.20°.
  • the above-mentioned B crystal form has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2 ⁇ angles: 7.06 ⁇ 0.20°, 14.23 ⁇ 0.20°, 19.33 ⁇ 0.20° and 20.91 ⁇ 0.20°, and can also be 17.16 ⁇ 0.20°, and/or 20.03 ⁇ 0.20°, and/or 23.91 ⁇ 0.20°, and/or 21.46 ⁇ 0.20°, and/or 25.15 ⁇ 0.20°, and/or 14.60 ⁇ 0.
  • the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2 ⁇ angles: 7.06 ⁇ 0.20°, 14.23 ⁇ 0.20°, 17.16 ⁇ 0.20°, 19.33 ⁇ 0.20°, 20.03 ⁇ 0.20°, 20.91 ⁇ 0.20°, 21.46 ⁇ 0.20° and 23.91 ⁇ 0.20°.
  • the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2 ⁇ angles: 7.06 ⁇ 0.20°, 14.23 ⁇ 0.20°, 14.60 ⁇ 0.20°, 17.16 ⁇ 0.20°, 19.33 ⁇ 0.20°, 20.03 ⁇ 0.20°, 20.91 ⁇ 0.20°, 21.46 ⁇ 0.20°, 23.91 ⁇ 0.20°, 24.49 ⁇ 0.20°, 25.15 ⁇ 0.20° and 27.14 ⁇ 0.20°.
  • the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2 ⁇ angles: 7.06 ⁇ 0.20°, 14.23 ⁇ 0.20°, 14.60 ⁇ 0.20°, 17.16 ⁇ 0.20°, 19.33 ⁇ 0.20°, 20.03 ⁇ 0.20°, 20.91 ⁇ 0.20°, 21.46 ⁇ 0.20°, 21.90 ⁇ 0.20°, 22.36 ⁇ 0.20°, 23.60 ⁇ 0.20°, 23.91 ⁇ 0.20°, 24.49 ⁇ 0.20°, 25.15 ⁇ 0.20°, 27.14 ⁇ 0.20° and 29.89 ⁇ 0.20°.
  • the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2 ⁇ angles: 7.06 ⁇ 0.20°, 9.40 ⁇ 0.20°, 10.83 ⁇ 0.20°, 12.39 ⁇ 0.20°, 14.23 ⁇ 0.20°, 14.60 ⁇ 0.20°, 15.31 ⁇ 0.20°, 15.99 ⁇ 0.20°, 16.24 ⁇ 0.20°, 17.16 ⁇ 0.20°, 18.96 ⁇ 0.20°, 19.33 ⁇ 0.20°, 20.03 ⁇ 0.20°, 20.91 ⁇ 0.20°, 21.46 ⁇ 0.20°.
  • the XRPD spectrum analysis data of the above-mentioned Form B is basically as shown in Table 2.
  • the XRPD spectrum of Form B of compound TM03 is basically as shown in FIG. 9 .
  • the B crystal form of compound TM03 is characterized by having any one of the following characteristics:
  • thermogravimetric analysis curve shows no weight loss at 30-200°C
  • the present invention also provides a method for preparing the B crystal form of compound TM03, comprising the following steps (a) to (c):
  • the present invention also provides a D crystal form of compound TM03, characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2 ⁇ angles: 3.55 ⁇ 0.20°, 18.57 ⁇ 0.20° and 20.63 ⁇ 0.20°;
  • the XRPD spectrum analysis data of the above-mentioned D crystal form is basically as shown in Table 4.
  • the XRPD spectrum of the D crystal form of compound TM03 is basically as shown in FIG. 13 .
  • the differential scanning calorimetry curve of the above-mentioned D crystal form has an endothermic peak starting point at 180.18°C ⁇ 5°C and 327.60°C ⁇ 5°C respectively; and has an exothermic peak starting point at 128.94°C ⁇ 5°C.
  • the DSC spectrum of the above-mentioned D crystal form is basically as shown in Figure 14.
  • thermogravimetric analysis curve of the above-mentioned D crystal form shows a weight loss of 2.861% at 165°C.
  • the TGA spectrum of the above-mentioned D crystal form is basically as shown in Figure 15.
  • the present invention also provides a method for preparing the D crystal form of compound TM03, comprising the following steps (a) to (c):
  • the present invention also provides the use of the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt in the preparation of drugs for treating diseases related to P2X3 inhibitors.
  • the present invention also provides the use of the crystalline form of compound TM03 in the preparation of drugs for treating diseases related to P2X3 inhibitors.
  • the P2X3 inhibitor-related disease refers to pain or chronic cough.
  • the P2X3 inhibitor-related disease refers to refractory cough and/or COVID-19-related cough.
  • the present invention also provides the following synthesis method:
  • the present invention also provides the following testing method:
  • Test Method 1 In vitro enzyme activity test of the compounds of the present invention
  • the IC 50 value was determined using 33 P isotope-labeled kinase activity assay (Reaction Biology Corp) to evaluate the inhibitory ability of the test compounds on P2X3 receptors.
  • Buffer conditions 20 mM Hepes (pH 7.5), 10 mM MgCl 2 , 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na vanadate (Na 3 VO 4 ), 2 mM DTT, 1% DMSO.
  • Test steps At room temperature, the test compound was dissolved in DMSO to prepare a 10mM solution for use.
  • the substrate was dissolved in a freshly prepared buffer, to which the kinase to be tested was added and mixed evenly.
  • the DMSO solution containing the test compound was added to the above mixed reaction solution using acoustic technology (Echo 550).
  • the concentration of the compound in the reaction solution was 10 ⁇ M, 2.50 ⁇ M, 0.62 ⁇ M, 0.156 ⁇ M, 39.1nM, 9.8nM, 2.4nM, 0.61nM, 0.15nM, 0.038nM or 3 ⁇ M, 1 ⁇ M, 0.333 ⁇ M, 0.111 ⁇ M, 37.0nM, 12.3nM, 4.12nM, 1.37nM, 0.457nM, 0.152nM.
  • 33 P-ATP activity 0.01 ⁇ Ci/ ⁇ L
  • the reaction solution was spotted on P81 ion exchange filter paper (Whatman #3698-915).
  • the kinase activity data were expressed as a comparison of the kinase activity of the test compound and the kinase activity of the blank group (containing only DMSO), and the IC 50 value was obtained by curve fitting using Prism4 software (GraphPad).
  • the compound of the present invention has excellent inhibitory activity on P2X3 receptors, has good exposure and bioavailability, can significantly reduce the number of coughs induced by ATP+citric acid in guinea pigs, and can significantly increase the latency of coughs induced by ATP+citric acid in guinea pigs.
  • pharmaceutically acceptable salt refers to salts of compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases.
  • base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts.
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent.
  • Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base or acid addition salts.
  • salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.
  • the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention.
  • Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention.
  • enantiomer or “optical isomer” refers to stereoisomers that are mirror images of one another.
  • cis-trans isomers or “geometric isomers” arises from the inability of a ring to rotate freely about double bonds or single bonds of ring carbon atoms.
  • diastereomer refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.
  • the key is a solid wedge. and dotted wedge key
  • a straight Line Key and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Denotes a solid wedge bond or dotted wedge key Or use a wavy line Represents a straight solid bond and straight dashed key
  • a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (in a double bond containing a nitrogen atom, a lone pair of electrons on the nitrogen atom is regarded as a substituent connected to it), if a wavy line is used between the atom on the double bond and its substituent in the compound, If connected, it means the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound.
  • formula (A) means that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2) or in the form of a mixture of two isomers of formula (A-1) and formula (A-2);
  • formula (B) means that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2) or in the form of a mixture of two isomers of formula (B-1) and formula (B-2).
  • formula (C) means that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2) or in the form of a mixture of two isomers of formula (C-1) and formula (C-2).
  • tautomer or “tautomeric form” refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and can rapidly convert into each other. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be achieved.
  • proton tautomers also called prototropic tautomers
  • keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one. For example and Interconversion between two tautomers.
  • the terms “enriched in one isomer”, “isomerically enriched”, “enriched in one enantiomer” or “enantiomerically enriched” mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
  • the term “isomer excess” or “enantiomeric excess” refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.
  • Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary. Wherein the resulting diastereomeric mixture is separated and the auxiliary groups are cleaved to provide the pure desired enantiomer.
  • a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art and then the pure enantiomers are recovered.
  • the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamates are generated from amines).
  • the compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound.
  • the compound may be labeled with a radioactive isotope, such as tritium ( 3H ), iodine-125 ( 125I ) or C-14 ( 14C ).
  • deuterated drugs may be formed by replacing hydrogen with heavy hydrogen. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing therapeutic effects, and extending the biological half-life of drugs. All isotopic composition changes of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.
  • substituted means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable.
  • oxygen it means that two hydrogen atoms are replaced.
  • Oxygen substitution does not occur on aromatic groups.
  • optionally substituted means that it may be substituted or not substituted, and unless otherwise specified, the type and number of the substituent can be arbitrary on the basis of chemical achievable.
  • any variable e.g., R
  • its definition at each occurrence is independent.
  • the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice.
  • substituents and/or variants thereof are permitted only if such combinations result in stable compounds.
  • linking group When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
  • substituent When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A.
  • substituent does not specify which atom it is connected to the substituted group through, the substituent can be bonded through any atom of it.
  • pyridyl as a substituent can be connected to the substituted group through any carbon atom on the pyridine ring.
  • connection direction is arbitrary.
  • the connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of the reading order from left to right to form Combinations of linkers, substituents, and/or variations thereof are permissible only if such combinations result in stable compounds.
  • C 1-4 alkyl is used to represent a straight or branched saturated hydrocarbon group consisting of 1 to 4 carbon atoms.
  • the C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc.; they can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine).
  • Examples of C 1-4 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), etc.
  • C 1-3 alkyl is used to represent a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms.
  • the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine).
  • Examples of C 1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.
  • C 1-4 alkoxy refers to those alkyl groups containing 1 to 4 carbon atoms connected to the rest of the molecule through an oxygen atom.
  • the C 1-4 alkoxy includes C 1-3 , C 1-2 , C 2-4 , C 4 and C 3 alkoxy, etc.
  • Examples of C 1-4 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), etc.
  • C 1-3 alkoxy refers to those alkyl groups containing 1 to 3 carbon atoms connected to the rest of the molecule through an oxygen atom.
  • the C 1-3 alkoxy includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy, etc.
  • Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.
  • C 1-3 haloalkyl refers to monohaloalkyl and polyhaloalkyl groups containing 1 to 3 carbon atoms.
  • the C 1-3 haloalkyl group includes C 1-2 , C 2-3 , C 3 , C 2 and C 1 haloalkyl groups, etc.
  • Examples of C 1-3 haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, pentachloroethyl, 3-bromopropyl, etc.
  • C 1-4 alkylamino refers to those alkyl groups containing 1 to 4 carbon atoms attached to the rest of the molecule through an amino group.
  • the C 1-4 alkylamino group includes C 1-3 , C 1-2 , C 2-4 , C 4 , C 3 and C 2 alkylamino groups, etc.
  • C 1-4 alkylamino groups include, but are not limited to, -NHCH 3 , -N(CH 3 ) 2 , -NHCH 2 CH 3 , -N(CH 3 )CH 2 CH 3 , -N(CH 2 CH 3 )(CH 2 CH 3 ), -NHCH 2 CH 2 CH 3 , -NHCH 2 (CH 3 ) 2 , -NHCH 2 CH 2 CH 2 CH 3 , and the like.
  • the terms “5-6 membered heteroaromatic ring” and “5-6 membered heteroaryl” of the present invention can be used interchangeably, and the term “5-6 membered heteroaryl” means a monocyclic group consisting of 5 to 6 ring atoms with a conjugated ⁇ electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. Wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2).
  • the 5-6 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom.
  • the 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl.
  • Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl) and 4H-1,2,4-triazolyl, etc
  • halo or halogen, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention.
  • the intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention.
  • the structure of the compound of the present invention can be confirmed by conventional methods known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art.
  • single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data of the cultured single crystal using a Bruker D8 venture diffractometer, the light source is CuK ⁇ radiation, and the scanning mode is: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.
  • SXRD single crystal X-ray diffraction
  • the solvent used in the present invention can be obtained from commercial sources.
  • the present invention uses the following abbreviations: DMSO represents dimethyl sulfoxide; EtOH represents ethanol; ACN represents acetonitrile; hr represents hour; EA represents ethyl acetate; MTBE represents methyl tert-butyl ether; THF represents tetrahydrofuran; and PE represents petroleum ether.
  • XRPD X-ray powder diffractometer
  • DSC differential scanning calorimeter
  • TGA Thermogravimetric analysis
  • ⁇ W% indicates the weight gain of the test sample at 25 ⁇ 1°C and 80 ⁇ 2%RH.
  • the hP2X3 cocrystal structure (PDB ID code: 5SVR) was used as the docking template for this binding mode prediction.
  • Maestro The protein preparation wizard module of version 2021-2 [1] added hydrogen atoms and used the OPLS4 force field.
  • the cocrystal structure was hydrogen bond optimized and the cocrystal small molecules were removed.
  • the water molecules other than water molecules and the overall energy optimization were first docked to determine the binding mode of the molecule S-600918 published by Shionogi.
  • the Glide [2] Receptor Grid Generation module was used to generate the center of mass of the hP2X3 antagonist A-317491 with a known cocrystal structure.
  • the docking grid was used and the Ligand Docking module was used to find the optimal binding mode of S-600918.
  • the pharmacophore and key protein-ligand interactions of S-600918 in this binding mode are similar to those of A-317491, which proves the reliability of the docking model.
  • the 3D structure of the newly designed molecules was generated using LigPrep [3] and energy minimized. Based on this docking model, the newly designed molecules were docked using the SP docking mode in Glide [2] .
  • the binding modes of compounds 1 to 5 are shown in Figures 1-5.
  • the compounds of the present invention have good binding with hP2X3 ion channels, and well reproduce the binding mode of Yangshen molecule S-600918: this series of molecules form salt bridges with Arg281, Lys299, and Lys63, and form hydrogen bonds with Asn279 and Ser275.
  • the 2-phenoxypyridine fragment inserts into the hydrophobic pocket to form ⁇ - ⁇ stacking with Phe174, the chlorobenzene fragment forms a cation-pi interaction with Lys65, and has an electrostatic interaction with Thr172.
  • the docking score of the newly designed molecule is close to or better than that of the Yangshen molecule.
  • the binding with the hP2X3 ion channel will effectively inhibit the overexcitation of airway sensory neurons and relieve cough symptoms.
  • Step 1 Synthesis of compound TM01_2
  • Step 1 Synthesis of compounds TM02 and TM03
  • TM01 (100 mg, 203.69 ⁇ mol) was subjected to chiral separation and separated by preparative SFC separation (column type: DAICEL CHIRALPAK IG (250 mm*30 mm, 10 ⁇ m); mobile phase: [CO 2 -EtOH]; EtOH%: 40%; separation time: 15 mins) to give compound TM02 (retention time: 7.145 min, ee value: 100%) and compound TM03 (retention time: 8.106 min, ee value: 97.14%).
  • Step 1 Synthesis of compound TM07_2
  • Step 1 Synthesis of compound TM08_2
  • TM08_5 (540 mg, 1.32 mmol) was added to tert-butyl alcohol (5 mL), followed by TM01_5 (368.83 mg, 1.98 mmol) and acetic acid (1.12 mL), stirred at 115°C for 12 hr, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL ⁇ 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent.
  • Step 1 Synthesis of compound TM09_1
  • Step 1 Synthesis of compound TM10_1
  • Step 1 Synthesis of compound TM11_1
  • TM08_5 100 mg, 244.53 ⁇ mol was added to tert-butyl alcohol (2 mL), followed by compound TM06_1 (80.94 mg, 366.80 ⁇ mol) and acetic acid (207.74 ⁇ L), stirred at 115°C for 12 hours, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL ⁇ 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent.
  • Step 1 Synthesis of compound TM12_1
  • TM13_1 (40.42 g, 199.77 mmol, 49.29 mL) was slowly added dropwise to tri-n-butylphosphine (100 g, 998.85 mmol, 108.58 mL) under nitrogen protection, and the reaction was stirred at 60°C for 2 hr.
  • TM13_7 (0.17 g, 320.15 ⁇ mol) and tetrahydrofuran (1.7 mL), methanol (1.7 mL), water (0.35 mL) were added to a pre-dried single-mouth bottle, followed by the addition of lithium hydroxide monohydrate (4 M, 656.31 ⁇ L) and the reaction was carried out at 50°C for 2 h. After the reaction was completed, 4 mL of water was added, and the mixture was extracted with ethyl acetate (5 mL ⁇ 3).
  • Tetraisopropyl titanium oxide (6.34 g, 22.29 mmol) and titanium tetrachloride (13.01 g, 68.59 mmol) were dissolved in dichloromethane (200 mL), cooled to 0°C, stirred for 10 min, and then diisopropylethylamine (12.08 g, 93.46 mmol) was added. The mixture was kept at 0°C and stirred for 20 min, and then compound TM14_1 (20 g, 85.74 mmol) was added.
  • Hydrogen peroxide (16.94 g, 149.41 mmol, 14.36 mL, 30% purity) was dissolved in tetrahydrofuran (150 mL) and then cooled to 0°C. After adding a solution of lithium hydroxide (2.09 g, 49.80 mmol) in water (50 mL) while maintaining 0°C, a solution of compound TM14_2 (15 g, 41.50 mmol) in tetrahydrofuran (50 mL) was added dropwise at 0°C and stirred for 2 hours at 0°C.
  • TM15_2 (3.8 g, 17.10 mmol) was dissolved in tetrahydrofuran (16 mL), iron (3.82 g, 68.40 mmol), ammonium chloride (3.66 g, 68.40 mmol) and water (12 mL) were added, and the nitrogen was replaced three times and stirred at 40°C for 48 hours.
  • the reaction solution was filtered, the solid was washed with ethyl acetate (30 mL), separated, and the liquid was extracted with ethyl acetate (30 mL ⁇ 3).
  • the organic phases were combined and dried over anhydrous sodium sulfate and then dried under reduced pressure to obtain TM15_3, MS m/z: 193.1 [M+1] + .
  • Step 1 Synthesis of compound TM16_1
  • TM01 75 mg, 152.77 ⁇ mol was dissolved in dichloromethane (3 mL), the reaction system was cooled to 0°C, oxalyl chloride (29.09 mg, 229.15 ⁇ mol) was added, and the reaction temperature did not exceed 10°C during the addition. Then a drop of N,N-dimethylformamide was added, and the reaction system was stirred at 20°C for 2 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. TM16_1 was obtained, and the crude product was directly used for the next step. MS m/z: 505.2[M+1] + .
  • TM16_1 (17.44 mg, 183.33 ⁇ mol) was dissolved in dichloromethane (0.5 mL), N, N-diisopropylethylamine (59.23 mg, 458.32 ⁇ mol) was added, the reaction system was cooled to 0°C, methylsulfonamide (77.82 mg, 152.77 ⁇ mol) dissolved in dichloromethane (0.5 mL) was added dropwise, the temperature was slowly raised to 20°C and stirred for 12 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure.
  • 1 H NMR 400MHz, DMSO-d 6 ) ⁇ ppm 11.80-11.89 (m, 1H) 8.13-8.17 (m, 1H) 8.12 (s, 1H) 7.82-7.92 (m, 1H) 7.46 (br s, 4H) 7.34-7.42 (m, 2H) 7.11-7.19 (m, 3H) 7.
  • Step 1 Synthesis of compound TM20_2
  • TM20_2 (60.00 mg, 111.74 ⁇ mol) was added to tetrahydrofuran (0.6 mL), methanol (0.6 mL) and water (0.2 mL), and lithium hydroxide monohydrate (8.03 mg, 335.21 ⁇ mol) was added, and stirred at 45°C for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (3 M), and ethyl acetate (10 mL ⁇ 3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent.
  • Step 1 Synthesis of compound TM21_2
  • TM21_2 (55 g, 274.71 mmol) was added to tetrahydrofuran (300 mL) and water (300 mL), and then lithium hydroxide (34.58 g, 824.12 mmol) was added, and the mixture was stirred at 25°C for 12 hours, and the solvent was removed by concentrating under reduced pressure, and then extracted with ethyl acetate (1000 mL). After collecting the aqueous phase, the solvent was removed by concentrating under reduced pressure, and the pH was adjusted to 3-4 with dilute hydrochloric acid (3 M), and the mixture was filtered, the filter cake was collected, and the solvent was removed by concentrating under reduced pressure.
  • TM21_3 (5 g, 19.43 mmol) was dissolved in dichloromethane (80 mL), and then p-chlorobenzyl bromide (3.99 g, 19.43 mmol) and diisopropylethylamine (5.02 g, 38.86 mmol) were added in sequence, and the reaction solution was stirred at room temperature (15°C) for 12 hours. After the reaction was completed, saturated aqueous ammonium chloride solution (50 mL) and dichloromethane (50 mL) were added to the reaction solution to separate the organic phase, and the organic phase was concentrated under reduced pressure to obtain a crude product.
  • TM21_4 (1 g, 2.62 mmol) was dissolved in tetrahydrofuran (5 mL), cooled to 0°C, and then sodium hydrogen (314.24 mg, 7.86 mmol, 60% purity) and methyl bromoacetate (440.65 mg, 2.88 mmol) were added.
  • the reaction solution was heated to room temperature 20°C and stirred for 2 hours.
  • saturated aqueous ammonium chloride solution (20 mL) was added dropwise to the reaction solution to quench the reaction, and then extracted with ethyl acetate (30 mL ⁇ 2), the organic phase was separated, and the organic phase was concentrated under reduced pressure to obtain a crude product.
  • TM21_5 (430 mg, 947.26 ⁇ mol) was added to pivalic acid (0.7 mL), and then TM01_5A (264.58 mg, 1.42 mmol) was added, and stirred at 130°C for 2 hr. After the reaction was completed, 5 mL of saturated sodium bicarbonate solution was slowly added to quench the reaction, and ethyl acetate (10 mL ⁇ 3) was used for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure.
  • Step 1 Synthesis of Compound TM22_1 and Compound TM23_1
  • Compound TM04_2 was separated by preparative SFC (chromatographic column: DAICELCHIRALCELOJ (250mm*30mm, 10 ⁇ m); mobile phase: [A: CO 2 -B: MeOH (0.1% NH 3 H 2 O)]; B%: 60%, isobaric elution mode), separation time: 3mins, to obtain compound TM22_1 (retention time: 1.441min, ee value 100%) and compound TM23_1 (retention time: 1.786min, ee value 99.76%).
  • TM22_1 (100.00 mg, 186.23 ⁇ mol) was dissolved in tetrahydrofuran (0.5 mL) and water (0.5 mL), and then lithium hydroxide (11.72 mg, 279.34 ⁇ mol) was added and stirred at 20°C for 2.5 hr. After the reaction was completed, the reaction solution was adjusted to pH 3-4 with 1M dilute hydrochloric acid, and the solid was precipitated, the filter cake was collected by filtration, and the solvent was removed by vacuum concentration.
  • TM22 was purified by preparative HPLC (chromatographic column: Phenomenex Luna C18 75*30mm*3 ⁇ m; mobile phase: [H 2 O (0.04% HCl)-ACN]; gradient: 20%-65% ACN, 8.0 min) and freeze-dried to obtain TM22. MSm/z: 509.1[M+1] + .
  • TM23_1 (100.00 mg, 186.23 ⁇ mol) was dissolved in tetrahydrofuran (0.5 mL) and water (0.5 mL), and then lithium hydroxide (11.72 mg, 279.34 ⁇ mol) was added and stirred at 20°C for 2.5 hr. After the reaction was completed, the pH of the reaction solution was adjusted to 3-4, and the solid was precipitated, and the filter cake was collected by filtration and concentrated under reduced pressure to remove the solvent.
  • TM23 was purified by preparative HPLC (chromatographic column: Phenomenex Luna C18 75*30mm*3 ⁇ m; mobile phase: [water (0.04% HCl)-ACN]; gradient: 20%-65% ACN, 8.0 min) and freeze-dried to obtain TM23. MS m/z: 509.1[M+1] + .
  • Step 1 Synthesis of compound TM24_1
  • TM24_1 400 mg, 769.26 ⁇ mol
  • tetrahydrofuran (6 mL)
  • methanol (6 mL)
  • water 2 mL
  • lithium hydroxide 129.12 mg, 3.08 mmol
  • the pH was adjusted to 1-2 with 1 M dilute hydrochloric acid, and then the residue was concentrated under reduced pressure. The residue was dissolved with 3 mL DMF and filtered for preparative separation.
  • Step 1 Synthesis of compound TM25_1
  • TM25_1 (100 mg, 186.23 ⁇ mol) was added to tetrahydrofuran (0.6 mL), methanol (0.6 mL) and water (0.2 mL), and lithium hydroxide monohydrate (13.38 mg, 558.68 ⁇ mol) was added, and stirred at 45°C for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (3 M), and ethyl acetate (10 mL ⁇ 3) was added for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent.
  • Example 32 Solid pre-stability test of Form B of compound TM03
  • Test substance compound of the present invention
  • Reference substance ⁇ -methyleneATP, AF-219
  • Solvent dimethyl sulfoxide (DMSO)
  • Agonist Weigh an appropriate amount of ⁇ -methylene ATP and prepare a 20mM stock solution with sterile water. Store at -80°C after aliquoting.
  • Inhibitor Weigh an appropriate amount of AF-219 and prepare a 10 mM stock solution with DMSO. Store at -20°C after aliquoting.
  • Working solution preparation design Before testing human P2X3 and P2X2/3 receptors, take out the agonist and inhibitor stock solutions and the test substance stock solutions.
  • the detection concentration of the agonist ⁇ -methylene ATP is 300 ⁇ M starting, 3-fold dilution, 9 concentrations, 2 replicates; the detection concentration of the inhibitor AF-219 is 10 ⁇ M starting, 3-fold dilution, 9 concentrations, 2 replicates; the test substance detection concentration is 1 ⁇ M or 100 ⁇ M starting, 3-fold dilution, 9 concentrations, 2 replicates; the test substance activates human P2X3 and P2X2/3 channels with the concentration of ⁇ -methyleneATPEC80.
  • the blank control is 0.5% DMSO.
  • Preparation of compound detection plate Set up the plate-making program on the ECHO555 liquid workstation. When making the plate, the final concentration of the compound plate needs to be set to 3 or 4 times the detection concentration.
  • the ECHO-specific LDV plate needs to first use the solvent DMSO or double distilled water to dilute the compound storage solution to 10mM, 1mM and 0.1mM, and make a compound concentration gradient plate. Then prepare the solvent DMSO to make the plate, so that the total volume of each well is replenished to 200nL, and ensure that the DMSO content is consistent in each well. At this time, the compound concentration is 600 times or 800 times the concentration to be tested.
  • hP2X cell lines are passaged twice a week at a dilution ratio of 1:3 or 1:4.
  • trypsin-EDTA solution When the cells reach 80% or more confluence in a T-75 flask, use 0.25% trypsin-EDTA solution to digest the cells for about 2-5 minutes until the cells can fall off naturally. Finally, transfer the cell suspension to another T-75 flask for subculture according to the dilution ratio.
  • Cell seeding After resuspending the cells as described above, the cell density and viability were determined by the fully automated cell counter Cell Countess. After calculation, the volume of the cell suspension was adjusted with complete culture medium, and hP2X3 cells were seeded at a density of 10,000 cells/well and hP2X2/3 cells at a density of about 18,000 cells/well (30 ⁇ L/well) on a PDL-coated black bottom transparent 384-well plate. The plate was placed in a humidified air controlled (5% CO 2 ) 37°C incubator for overnight culture.
  • 5% CO 2 5% CO 2
  • FLIPR plate reading Take out the cell plate cultured overnight from the incubator, discard the culture medium and add Ca6 dye (Molecular Devices, USA). Add 25 ⁇ L Ca6 to each well and incubate for 2 hours. Finally, put the cell plate, compound plate, agonist plate and FLIPR matching 384-well plate tip into FLIPR PENTA (Molecular Devices, USA), set the calcium flow detection excitation light to 470-515nm, the emission light to 515-575nm, correct the background signal window to 700-800RFU, and run the FLIPR program.
  • the specific procedure is to read the baseline for 60 seconds, then the FLIPR tip draws 12.5 ⁇ L from the 3-fold concentration compound plate and adds it to the cell plate, and then draws 12.5 ⁇ L from the 4-fold concentration stimulation plate and adds it to the cell plate after 90 seconds.
  • the data recording lasts for at least 3 minutes, and the sampling rate is set to 1Hz.
  • the raw data is exported and analyzed as the maximum value of ⁇ F/F after the addition of the agonist.
  • Quality control The average value and standard deviation are used for duplicate calculation. The window is obtained by dividing the maximum signal value by the minimum signal value. If it exceeds 2.5 times, the test is acceptable. The Z factor is used as a performance indicator, and if it exceeds 0.5, it is acceptable.
  • the compounds of the present invention have excellent inhibitory activity on P2X3 receptors.
  • Hartley guinea pigs male, 400-500 g, 6-9 weeks old, Beijing Weitong Lihua
  • This project uses 4 male Hartley guinea pigs. They are weighed before administration and the dosage is calculated based on their body weight. The rats are then divided into two groups. Two Hartley guinea pigs in one group were intravenously injected with the drug at a dose of 1 mg/kg and a concentration of 0.5 mg/mL; another group of two Hartley guinea pigs were orally administered with a dose of 3 mg/kg and a concentration of 0.5 mg/mL;
  • Plasma samples were stored in a -80°C refrigerator before analysis.
  • the compounds of the present invention have good exposure and bioavailability.
  • the therapeutic effect of the target compound on guinea pig cough was investigated, and animal experiments were conducted on the effect on ATP+citric acid-induced guinea pig cough, which proved that the test drug could reduce the number of coughs.
  • Citric acid modeling group Citric acid modeling group, ATP+citric acid modeling, ATP+citric acid modeling and administration group 1, ATP+citric acid modeling and administration group 2, ATP+citric acid modeling and administration group 3, ATP+citric acid modeling and administration group 4, number of animals: 8/group.
  • Citric acid and ATP+citric acid modeling were performed according to the grouping method.
  • citric acid modeling group citric acid was nebulized for 10 minutes, and the number of guinea pig coughs was counted after 10 minutes of citric acid nebulization and 5 and 10 minutes of observation after stopping nebulization;
  • ATP+citric acid modeling group ATP was nebulized for 2 minutes and citric acid was nebulized for 10 minutes, and the number of guinea pig coughs was counted after 10 minutes of nebulization and 5 and 10 minutes of observation after stopping nebulization.
  • Administration time 2 hours before modeling, by intragastric administration.
  • Citric acid modeling Before modeling, the guinea pigs were placed in the atomization drug delivery box for 10 minutes to adapt. After the adaptation was completed, the drug delivery device was adjusted to atomize 0.1 M citric acid at an atomization rate of 0.6 mL/min, and the atomization was stopped after 10 minutes of atomization.
  • ATP+citric acid modeling Before modeling, the guinea pigs were placed in the nebulizer drug delivery box to adapt for 10 minutes. The drug delivery device was adjusted in the last 2 minutes to administer 10 ⁇ M ATP by nebulization at a nebulization rate of 0.6 mL/min. After the ATP nebulization was completed, 0.1 M citric acid was administered by nebulization at a nebulization rate of 0.6 mL/min. The nebulization was stopped after 10 minutes of nebulization.
  • the compound of the present invention can significantly reduce the number of coughs induced by ATP+citric acid in guinea pigs, and can significantly increase the latency of coughs induced by ATP+citric acid in guinea pigs.

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Abstract

Disclosed are a pyrimidinone derivative and an application thereof, and specifically disclosed is a compound as shown in formula (III), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

Description

嘧啶酮衍生物及其在药学上的应用Pyrimidone derivatives and their pharmaceutical applications

本申请主张如下优先权：This application claims the following priority:

申请号：CN202310120433.X，申请日：2023年02月15日；Application number: CN202310120433.X, application date: February 15, 2023;

申请号：CN202310213602.4，申请日：2023年03月07日；Application number: CN202310213602.4, application date: March 7, 2023;

申请号：CN202310257095.4，申请日：2023年03月16日；Application number: CN202310257095.4, application date: March 16, 2023;

申请号：CN202310354516.5，申请日：2023年04月04日；Application number: CN202310354516.5, application date: April 4, 2023;

申请号：CN202310361117.1，申请日：2023年04月06日；Application number: CN202310361117.1, application date: April 6, 2023;

申请号：CN202310450274.X，申请日：2023年04月24日；Application number: CN202310450274.X, application date: April 24, 2023;

申请号：CN202310630849.6，申请日：2023年05月30日；Application number: CN202310630849.6, application date: May 30, 2023;

申请号：CN202310677346.4，申请日：2023年06月08日。Application number: CN202310677346.4, application date: June 8, 2023.

技术领域Technical Field

本发明涉及一类嘧啶酮衍生物及其应用，具体涉及式(III)所示化合物、其立体异构体或其药学上可接受的盐。The present invention relates to a class of pyrimidone derivatives and applications thereof, and in particular to a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

背景技术Background Art

嘌呤受体是以三磷酸腺苷(adenosine triphosphate，ATP)为配体的离子通道型受体。目前共发现了多种嘌呤能受体亚型，包括7种离子通道型受体(ionotropic receptors P2X)、9种代谢型受体(metabotropic receptors P2Y)和4种腺苷受体。其中，嘌呤能离子通道型受体3(purinergic ligandgated ion channel 3 receptor，P2X3受体)具有独特且重要的生理功能。P2X3选择性地表达于初级感觉神经元，介导信号转导从而调控多种生理功能。ATP与P2X3受体结合后可将其激活，使得细胞膜对Na+、K+、Ca2+通透性增加，尤以Ca2+的通透性改变最为明显。当机体受到伤害或神经损伤可后释放大量ATP，激活突触前膜P2X3受体，引起大量Ca2+内流，细胞内钙浓度增加激活蛋白激酶A(protein kinase A，PKA)、蛋白激酶C(protein kinase C，PKC)，使得PKA、PKC磷酸化，同时促进了谷氨酸释放，进一步激活N-甲基-D-天门冬胺酸受体(N-methyl-D-aspartic acid receptor，NMDA)，导致兴奋性突触后电流的产生，引起中枢敏感化。研究表明，P2X3受体表达上调可导致痛敏形成，参与疼痛的信号传递，且P2X3受体可介导炎性疼痛、神经性疼痛、癌性疼痛等的产生。目前P2X3抑制剂的公开体内研究用于治疗咳嗽和疼痛等疾病，新型P2X3抑制剂的发现和应用具有广阔的前景，对于咳嗽和疼痛等疾病的治疗具有重要意义。Purinergic receptors are ion channel receptors with adenosine triphosphate (ATP) as ligand. Currently, a variety of purinergic receptor subtypes have been discovered, including 7 ion channel receptors (ionotropic receptors P2X), 9 metabotropic receptors (metabotropic receptors P2Y) and 4 adenosine receptors. Among them, purinergic ligandgated ion channel 3 receptor (P2X3 receptor) has unique and important physiological functions. P2X3 is selectively expressed in primary sensory neurons, mediating signal transduction and regulating a variety of physiological functions. ATP can activate P2X3 receptors after binding to them, increasing the permeability of the cell membrane to Na + , K + , and Ca 2+ , especially the permeability change of Ca 2+ is the most obvious. When the body is injured or nerves are damaged, a large amount of ATP can be released, activating the presynaptic P2X3 receptor, causing a large amount of Ca 2+ influx, and the increase in intracellular calcium concentration activates protein kinase A (PKA) and protein kinase C (PKC), causing PKA and PKC phosphorylation, while promoting the release of glutamate, further activating N-methyl-D-aspartic acid receptor (NMDA), leading to the generation of excitatory postsynaptic currents, causing central sensitization. Studies have shown that upregulation of P2X3 receptor expression can lead to the formation of pain sensitivity, participate in the signal transmission of pain, and P2X3 receptors can mediate the generation of inflammatory pain, neuropathic pain, cancer pain, etc. At present, the public in vivo research of P2X3 inhibitors is used to treat diseases such as cough and pain. The discovery and application of new P2X3 inhibitors have broad prospects and are of great significance for the treatment of diseases such as cough and pain.

发明内容Summary of the invention

本发明提供了式(III)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention provides a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

其中，in,

R1选自H、D、卤素、CN、C1-3烷基或C1-3卤代烷基；R 1 is selected from H, D, halogen, CN, C 1-3 alkyl or C 1-3 haloalkyl;

各R2分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、-CH2OH、环丙基、C1-3烷基和C1-3烷氧基，所述环丙基、C1-3烷基和C1-3烷氧基任选被1、2或3个Ra取代； each R 2 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , -CH 2 OH, cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy, wherein the cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 Ra ;

各R3分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基，所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rb取代；each R 3 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R b ;

各R4分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基，所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rc取代；each R 4 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R c ;

R5和R6各自独立地选自H、D、F、CN和C1-4烷基；R 5 and R 6 are each independently selected from H, D, F, CN and C 1-4 alkyl;

或者，R5和R6及相连的碳原子形成环丙基；Alternatively, R 5 and R 6 and the carbon atom to which they are connected form a cyclopropyl group;

R7和R8各自独立地选自H、D、F、OH、CN和C1-4烷基；R 7 and R 8 are each independently selected from H, D, F, OH, CN and C 1-4 alkyl;

或者，R7和R8及相连的碳原子形成环丙基、环丁基或氧杂环丁基；Alternatively, R7 and R8 and the carbon atoms to which they are connected form a cyclopropyl group, a cyclobutyl group or an oxetanyl group;

R9选自OH、C1-4烷氧基、C1-4烷氨基和所述C1-4烷氧基和C1-4烷氨基各自独立地任选被1、2或3个选自F、Cl、Br、I和OH的取代基取代；R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and The C 1-4 alkoxy and C 1-4 alkylamino groups are each independently optionally substituted by 1, 2 or 3 substituents selected from F, Cl, Br, I and OH;

各Ra分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each Ra is independently selected from D, F, Cl, Br, I, CN, NH2 and OH;

各Rb分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R b is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;

各Rc分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R c is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;

n选自0、1、2、3和4；n is selected from 0, 1, 2, 3 and 4;

m选自0、1、2、3和4；m is selected from 0, 1, 2, 3 and 4;

r选自0、1、2、3和4；r is selected from 0, 1, 2, 3 and 4;

t选自1、2和3；t is selected from 1, 2 and 3;

环A1选自苯基和5-10元杂芳基；Ring A1 is selected from phenyl and 5-10 membered heteroaryl;

环A2选自苯基和5-6元杂芳基；Ring A2 is selected from phenyl and 5-6 membered heteroaryl;

环A3选自苯基和5-6元杂芳基；Ring A3 is selected from phenyl and 5-6 membered heteroaryl;

E1不存在，或选自NR10；E 1 is absent or selected from NR 10 ;

R10选自H和C1-4烷基；R 10 is selected from H and C 1-4 alkyl;

当t为2，且E1不存在时，R7、R8不全是H。When t is 2 and E1 does not exist, R7 and R8 are not all H.

本发明提供了式(III)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention provides a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

其中，in,

R1选自H、D、卤素、CN、C1-3烷基或C1-3卤代烷基；R 1 is selected from H, D, halogen, CN, C 1-3 alkyl or C 1-3 haloalkyl;

各R2分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、-CH2OH、环丙基、C1-3烷基和C1-3烷氧基，所述环丙基、C1-3烷基和C1-3烷氧基任选被1、2或3个Ra取代；each R 2 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , -CH 2 OH, cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy, wherein the cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 Ra ;

各R3分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基，所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rb取代；each R 3 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R b ;

各R4分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基， Each R 4 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy,

所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rc取代；The C 1-3 alkyl and C 1-3 alkoxy groups are optionally substituted by 1, 2 or 3 R c ;

R5和R6各自独立地选自H、D、F、CN和C1-4烷基；R 5 and R 6 are each independently selected from H, D, F, CN and C 1-4 alkyl;

或者，R5和R6及相连的碳原子形成环丙基；Alternatively, R 5 and R 6 and the carbon atom to which they are connected form a cyclopropyl group;

R7和R8各自独立地选自H、D、F、OH、CN和C1-4烷基；R 7 and R 8 are each independently selected from H, D, F, OH, CN and C 1-4 alkyl;

或者，R7和R8及相连的碳原子形成环丙基、环丁基或氧杂环丁基；Alternatively, R7 and R8 and the carbon atoms to which they are connected form a cyclopropyl group, a cyclobutyl group or an oxetanyl group;

R9选自OH、C1-4烷氧基、C1-4烷氨基和所述C1-4烷氧基和C1-4烷氨基各自独立地任选被1、2或3个选自F、Cl、Br、I和OH的取代基取代；R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and The C 1-4 alkoxy and C 1-4 alkylamino groups are each independently optionally substituted by 1, 2 or 3 substituents selected from F, Cl, Br, I and OH;

各Ra分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each Ra is independently selected from D, F, Cl, Br, I, CN, NH2 and OH;

各Rb分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R b is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;

各Rc分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R c is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;

n选自0、1、2、3和4；n is selected from 0, 1, 2, 3 and 4;

m选自0、1、2、3和4；m is selected from 0, 1, 2, 3 and 4;

r选自0、1、2、3和4；r is selected from 0, 1, 2, 3 and 4;

t选自1、2和3；t is selected from 1, 2 and 3;

环A1选自苯基和5-6元杂芳基；Ring A1 is selected from phenyl and 5-6 membered heteroaryl;

环A2选自苯基和5-6元杂芳基；Ring A2 is selected from phenyl and 5-6 membered heteroaryl;

环A3选自苯基和5-6元杂芳基；Ring A3 is selected from phenyl and 5-6 membered heteroaryl;

E1不存在，或选自NR10；E 1 is absent or selected from NR 10 ;

R10选自H和C1-4烷基；R 10 is selected from H and C 1-4 alkyl;

当t为2，且E1不存在时，R7、R8不全是H。When t is 2 and E1 does not exist, R7 and R8 are not all H.

本发明提供了式(III)所示化合物、其立体异构体或其药学上可接受的盐，
The present invention provides a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

其中，in,

R1选自H、D、卤素、CN、C1-3烷基或C1-3卤代烷基；R 1 is selected from H, D, halogen, CN, C 1-3 alkyl or C 1-3 haloalkyl;

各R2分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、-CH2OH、环丙基、C1-3烷基和C1-3烷氧基，所述环丙基、C1-3烷基和C1-3烷氧基任选被1、2或3个Ra取代；each R 2 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , -CH 2 OH, cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy, wherein the cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 Ra ;

各R3分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基，所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rb取代；each R 3 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R b ;

各R4分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基，所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rc取代；each R 4 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R c ;

R5和R6各自独立地选自H、D、F、CN和甲基；R 5 and R 6 are each independently selected from H, D, F, CN and methyl;

或者，R5和R6及相连的碳原子形成环丙基； Alternatively, R 5 and R 6 and the carbon atom to which they are connected form a cyclopropyl group;

R7和R8各自独立地选自H、D、F、OH、CN和甲基；R 7 and R 8 are each independently selected from H, D, F, OH, CN and methyl;

或者，R7和R8及相连的碳原子形成环丙基、环丁基或氧杂环丁基；Alternatively, R7 and R8 and the carbon atoms to which they are connected form a cyclopropyl group, a cyclobutyl group or an oxetanyl group;

R9选自OH、C1-4烷氧基和C1-4烷氨基，所述C1-4烷氧基和C1-4烷氨基各自独立地任选被1、2或3个选自F和OH的取代基取代；R 9 is selected from OH, C 1-4 alkoxy and C 1-4 alkylamino, wherein the C 1-4 alkoxy and C 1-4 alkylamino are each independently optionally substituted by 1, 2 or 3 substituents selected from F and OH;

各Ra分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each Ra is independently selected from D, F, Cl, Br, I, CN, NH2 and OH;

各Rb分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R b is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;

各Rc分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R c is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH;

n选自0、1、2、3和4；n is selected from 0, 1, 2, 3 and 4;

m选自0、1、2、3和4；m is selected from 0, 1, 2, 3 and 4;

r选自0、1、2、3和4；r is selected from 0, 1, 2, 3 and 4;

t选自1、2和3；t is selected from 1, 2 and 3;

环A1选自苯基和5-6元杂芳基；Ring A1 is selected from phenyl and 5-6 membered heteroaryl;

环A2选自苯基和5-6元杂芳基；Ring A2 is selected from phenyl and 5-6 membered heteroaryl;

环A3选自苯基和5-6元杂芳基；Ring A3 is selected from phenyl and 5-6 membered heteroaryl;

E1不存在，或选自NR10；E 1 is absent or selected from NR 10 ;

R10选自H和C1-4烷基。R 10 is selected from H and C 1-4 alkyl.

在本发明的一些方案中，上述R1选自H、D、F、Cl和甲基，其他变量如本发明所定义。In some embodiments of the present invention, the above R 1 is selected from H, D, F, Cl and methyl, and other variables are as defined in the present invention.

在本发明的一些方案中，上述各R2分别独立地选自H、D、F、Cl、CN、甲基和甲氧基，其他变量如本发明所定义。In some embodiments of the present invention, each R 2 mentioned above is independently selected from H, D, F, Cl, CN, methyl and methoxy, and other variables are as defined in the present invention.

在本发明的一些方案中，上述各R4分别独立地选自H、D、F、Cl和甲基，其他变量如本发明所定义。In some embodiments of the present invention, each R 4 mentioned above is independently selected from H, D, F, Cl and methyl, and other variables are as defined in the present invention.

在本发明的一些方案中，上述R5和R6各自独立地选自H、D、F和甲基，其他变量如本发明所定义。In some embodiments of the present invention, R 5 and R 6 are each independently selected from H, D, F and methyl, and other variables are as defined in the present invention.

在本发明中，当R7和R8及相连的碳原子形成环时，与R7和R8相连的碳原子可以是同一原子也可以是不同的原子。In the present invention, when R7 and R8 and the carbon atoms connected to them form a ring, the carbon atoms connected to R7 and R8 may be the same atom or different atoms.

在本发明的一些方案中，上述选自-CH2-、-(CH2)2-、-CH(CH3)-、-C(CH3)2-、-(CH2)3-、 -CH2CH(CH3)-、-CH2C(CH3)2-和-CH2CH(F)-，其他变量如本发明所定义。In some embodiments of the present invention, the above Selected from -CH 2 -, -(CH 2 ) 2 -, -CH(CH 3 )-, -C(CH 3 ) 2 -, -(CH 2 ) 3 -, -CH2CH ( CH3 )-, -CH2C ( CH3 ) 2- and -CH2CH (F)-, and other variables are as defined herein.

在本发明的一些方案中，上述选自-CH2-、-(CH2)2-、-CH(CH3)-、-(CH2)3-、 -CH2CH(CH3)-、-CH2C(CH3)2-和-CH2CH(F)-，其他变量如本发明所定义。In some embodiments of the present invention, the above is selected from -CH 2 -, -(CH 2 ) 2 -, -CH(CH 3 )-, -(CH 2 ) 3 -, -CH2CH ( CH3 )-, -CH2C ( CH3 ) 2- and -CH2CH (F)-, and other variables are as defined herein.

在本发明的一些方案中，上述选自-CH2-、-CH(CH3)-、-CH2CH(CH3)-和-CH2C(CH3)2-，其他变量如本发明所定义。In some embodiments of the present invention, the above Selected from -CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )- and -CH 2 C(CH 3 ) 2 -, and other variables are as defined herein.

在本发明的一些方案中，上述选自-CH2-和-CH2CH(CH3)-，其他变量如本发明所定义。 In some embodiments of the present invention, the above is selected from -CH 2 - and -CH 2 CH(CH 3 )-, and the other variables are as defined herein.

在本发明的一些方案中，上述选自-CH2CH(CH3)-，其他变量如本发明所定义。In some embodiments of the present invention, the above is selected from -CH 2 CH(CH 3 )-, and the other variables are as defined herein.

在本发明的一些方案中，上述R9选自OH、C1-4烷氧基、C1-4烷氨基和所述C1-4烷氧基和C1-4烷氨基各自独立地任选被1、2或3个F取代，其他变量如本发明所定义。In some embodiments of the present invention, the above R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and The C 1-4 alkoxy and C 1-4 alkylamino are each independently optionally substituted by 1, 2 or 3 F, and other variables are as defined herein.

在本发明的一些方案中，上述R9选自OH、C1-4烷氧基、C1-4烷氨基和所述C1-4烷氧基和C1-4烷氨基各自独立地任选被1、2或3个F取代，其他变量如本发明所定义。In some embodiments of the present invention, the above R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and The C 1-4 alkoxy and C 1-4 alkylamino are each independently optionally substituted by 1, 2 or 3 F, and other variables are as defined herein.

在本发明的一些方案中，上述R9选自OH、甲氧基、甲氨基、乙氨基和所述甲氧基、甲氨基、乙氨基各自独立地任选被1、2或3个F取代，其他变量如本发明所定义。In some embodiments of the present invention, the above R 9 is selected from OH, methoxy, methylamino, ethylamino and The methoxy group, methylamino group and ethylamino group are each independently optionally substituted with 1, 2 or 3 F groups, and other variables are as defined in the present invention.

在本发明的一些方案中，上述R9选自OH、OCH3、NHS(O)2CH3、NHCH3、NHCH2CHF2和NHCH2CF3，其他变量如本发明所定义。In some embodiments of the present invention, the above R 9 is selected from OH, OCH 3 , NHS(O) 2 CH 3 , NHCH 3 , NHCH 2 CHF 2 and NHCH 2 CF 3 , and other variables are as defined in the present invention.

在本发明的一些方案中，上述R9选自OH，其他变量如本发明所定义。In some embodiments of the present invention, the above R 9 is selected from OH, and other variables are as defined in the present invention.

在本发明的一些方案中，上述环A1选自苯基、 其他变量如本发明所定义。In some embodiments of the present invention, the ring A1 is selected from phenyl, Other variables are as defined in the present invention.

在本发明的一些方案中，上述环A1选自苯基、 其他变量如本发明所定义。In some embodiments of the present invention, the ring A1 is selected from phenyl, Other variables are as defined in the present invention.

在本发明的一些方案中，上述环A1选自其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述环A1选自其他变量如本发明所定义。 In some embodiments of the present invention, the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述环A1选自其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述环A1选自其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned ring A1 is selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述结构单元选自 其他变量如本发明所定义。In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述结构单元选自其他变量如本发明所定义。In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述结构单元选自其他变量如本发明所定义。In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

在本发明的一些方案中，上述t选自1，其他变量如本发明所定义。In some embodiments of the present invention, the above t is selected from 1, and other variables are as defined in the present invention.

在本发明的一些方案中，上述t选自2，其他变量如本发明所定义。In some embodiments of the present invention, the above t is selected from 2, and other variables are as defined in the present invention.

在本发明的一些方案中，上述t选自3，其他变量如本发明所定义。In some embodiments of the present invention, the above t is selected from 3, and other variables are as defined in the present invention.

在本发明的一些方案中，上述r选自0、1和2，其他变量如本发明所定义。In some embodiments of the present invention, the above r is selected from 0, 1 and 2, and other variables are as defined in the present invention.

在本发明的一些方案中，上述r选自1，其他变量如本发明所定义。In some embodiments of the present invention, the above r is selected from 1, and other variables are as defined in the present invention.

在本发明的一些方案中，上述环A2选自苯基，其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned ring A2 is selected from phenyl, and other variables are as defined in the present invention.

在本发明的一些方案中，上述环A3选自苯基，其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned ring A3 is selected from phenyl, and other variables are as defined in the present invention.

本发明的一些方案中，上述化合物、其立体异构体或其药学上可接受的盐，其化合物选自式(VII-1)和(III-1)所示结构：
In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt, the compound is selected from the structure shown in formula (VII-1) and (III-1):

其中，T1选自CH和N；R1、R2、R3、R4、R5、R6、R7、R8、R9、m、n、r和t如本发明所定义。wherein T1 is selected from CH and N; R1 , R2 , R3 , R4 , R5 , R6 , R7 , R8 , R9 , m, n, r and t are as defined in the present invention.

本发明的一些方案中，上述化合物、其立体异构体或其药学上可接受的盐，其化合物选自式(I-1)和(III-1)所示结构：
In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt, the compound is selected from the structure shown in formula (I-1) and (III-1):

其中，R1、R2、R3、R4、R5、R6、R7、R8、R9、m、n、r和t如本发明所定义。wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , m, n, r and t are as defined in the present invention.

本发明的一些方案中，上述化合物、其立体异构体或其药学上可接受的盐，其选自式(P-1)和(P-2)所示化合物：
In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compounds represented by formula (P-1) and (P-2):

其中，in,

T1选自CH和N； T1 is selected from CH and N;

E2选自NH和CH2；E 2 is selected from NH and CH 2 ;

R1、R2、R3、R4、R5、R6、R7、R8、R9、m、n、r和t如本发明所定义。 R1 , R2 , R3 , R4 , R5 , R6 , R7 , R8 , R9 , m, n, r and t are as defined herein.

本发明还有一些方案是由上述各变量任意组合而来。 Some other solutions of the present invention are obtained by arbitrarily combining the above variables.

本发明还提供下述化合物、其立体异构体或其药学上可接受的盐：





The present invention also provides the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:





本发明还提供下述化合物、其立体异构体或其药学上可接受的盐：




The present invention also provides the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:




本发明还提供下述化合物、其立体异构体或其药学上可接受的盐：




The present invention also provides the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:




本发明还提供化合物TM03的A晶型，其特征在于，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：13.69±0.20°，15.32±0.20°，17.30±0.20°，19.41±0.20°和21.45±0.20°；
The present invention also provides a crystalline form A of compound TM03, characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 13.69±0.20°, 15.32±0.20°, 17.30±0.20°, 19.41±0.20° and 21.45±0.20°;

本发明的一些方案中，上述A晶型，其特征在于，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：9.44±0.20°，13.69±0.20°，15.32±0.20°，17.30±0.20°，17.85±0.20°，19.41±0.20°，21.45±0.20°和21.73±0.20°。 In some embodiments of the present invention, the above-mentioned crystal form A is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 9.44±0.20°, 13.69±0.20°, 15.32±0.20°, 17.30±0.20°, 17.85±0.20°, 19.41±0.20°, 21.45±0.20° and 21.73±0.20°.

本发明的一些方案中，上述A晶型，其特征在于，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.57±0.20°，9.44±0.20°，13.69±0.20°，15.32±0.20°，17.30±0.20°，17.85±0.20°，18.55±0.20°，19.41±0.20°，20.24±0.20°，20.44±0.20°，21.45±0.20°，21.73±0.20°和23.48±0.20°。In some embodiments of the present invention, the above-mentioned crystal form A is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.57±0.20°, 9.44±0.20°, 13.69±0.20°, 15.32±0.20°, 17.30±0.20°, 17.85±0.20°, 18.55±0.20°, 19.41±0.20°, 20.24±0.20°, 20.44±0.20°, 21.45±0.20°, 21.73±0.20° and 23.48±0.20°.

本发明的一些方案中，上述A晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.57±0.20°，8.441±0.20°，9.44±0.20°，10.704±0.20°，12.12±0.20°，13.091±0.20°，13.687±0.20°，15.323±0.20°，16.14±0.20°，16.977±0.20°，17.304±0.20°，17.847±0.20°，18.55±0.20°，19.412±0.20°，20.24±0.20°，20.439±0.20°，21.449±0.20°，21.73±0.20°，22.833±0.20°，23.482±0.20°，24.648±0.20°，25.297±0.20°，25.594±0.20°，26.43±0.20°，27.141±0.20°，27.498±0.20°，27.932±0.20°，28.515±0.20°和30.231±0.20°。In some embodiments of the present invention, the above-mentioned crystal form A has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 7.57±0.20°, 8.441±0.20°, 9.44±0.20°, 10.704±0.20°, 12.12±0.20°, 13.091±0.20°, 13.687±0.20°, 15.323±0.20°, 16.14±0.20°, 16.977±0.20°, 17.304±0.20°, 17.847±0.20°, 18.55±0.20°, 1 9.412±0.20°, 20.24±0.20°, 20.439±0.20°, 21.449±0.20°, 21.73±0.20°, 22.833±0.20°, 23.482±0.20°, 24.648±0.20°, 25.297±0.20°, 25.594±0.20°, 26.43±0.20°, 27.141±0.20°, 27.498±0.20°, 27.932±0.20°, 28.515±0.20° and 30.231±0.20°.

本发明的一些方案中，上述A晶型的XRPD图谱解析数据基本上如表1所示。In some embodiments of the present invention, the XRPD spectrum analysis data of the above-mentioned Form A is basically as shown in Table 1.

表1化合物TM03的A晶型的XRPD图谱解析数据
Table 1 XRPD spectrum analysis data of Form A of compound TM03

本发明中，化合物TM03的A晶型，其XRPD图谱基本上如图6所示。In the present invention, the XRPD spectrum of Form A of compound TM03 is substantially as shown in FIG6 .

在本发明的一些方案中，上述A晶型的差示扫描量热曲线在111.1℃±5℃处具有吸热峰的起始点；176.3℃±5℃处具有放热峰的起始点。In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned crystal form A has a starting point of an endothermic peak at 111.1°C±5°C; and a starting point of an exothermic peak at 176.3°C±5°C.

在本发明的一些方案中，上述A晶型的DSC图谱基本上如图7所示。In some embodiments of the present invention, the DSC spectrum of the above-mentioned Form A is substantially as shown in FIG. 7 .

在本发明的一些方案中，上述A晶型的热重分析曲线在150℃时失重达9.325％。In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned crystal form A shows a weight loss of 9.325% at 150°C.

在本发明的一些方案中，上述A晶型的TGA图谱基本上如图8所示。In some embodiments of the present invention, the TGA spectrum of the above-mentioned crystal form A is basically as shown in Figure 8.

本发明还提供化合物TM03的A晶型的制备方法，其包括如下(a)～(c)的步骤：The present invention also provides a method for preparing the A crystal form of compound TM03, which comprises the following steps (a) to (c):

(a)将化合物TM03加入乙酸乙酯成澄清液，然后加入石油醚，有固体析出；(a) Compound TM03 was added to ethyl acetate to form a clear solution, and then petroleum ether was added to precipitate a solid;

(b)20～30℃下搅拌12小时；(b) stirring at 20-30° C. for 12 hours;

(c)过滤后干燥8～16小时。(c) After filtering, dry for 8 to 16 hours.

本发明还提供化合物TM03的B晶型，其特征在于，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°，14.23±0.20°，19.33±0.20°和20.91±0.20°；
The present invention also provides a crystal form B of compound TM03, characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 19.33±0.20° and 20.91±0.20°;

本发明的一些方案中，上述B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°，14.23±0.20°，19.33±0.20°，20.91±0.20°和21.46±0.20°。In some embodiments of the present invention, the above-mentioned B crystal form has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 19.33±0.20°, 20.91±0.20° and 21.46±0.20°.

本发明的一些方案中，上述B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°，14.23±0.20°，19.33±0.20°和20.91±0.20°，还可以在17.16±0.20°，和/或20.03±0.20°，和/或23.91±0.20°，和/或21.46±0.20°，和/或25.15±0.20°，和/或14.60±0.20°，和/或27.14±0.20°，和/或24.48±0.20°，和/或23.60±0.20°，和/或22.36±0.20°，和/或21.90±0.20°，和/或29.89±0.20°，和/或26.66±0.20°，和/或15.99±0.20°，和/或24.88±0.20°，和/或18.96±0.20°，和/或9.40±0.20°处有特征衍射峰。In some embodiments of the present invention, the above-mentioned B crystal form has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 19.33±0.20° and 20.91±0.20°, and can also be 17.16±0.20°, and/or 20.03±0.20°, and/or 23.91±0.20°, and/or 21.46±0.20°, and/or 25.15±0.20°, and/or 14.60±0. There are characteristic diffraction peaks at 20°, and/or 27.14±0.20°, and/or 24.48±0.20°, and/or 23.60±0.20°, and/or 22.36±0.20°, and/or 21.90±0.20°, and/or 29.89±0.20°, and/or 26.66±0.20°, and/or 15.99±0.20°, and/or 24.88±0.20°, and/or 18.96±0.20°, and/or 9.40±0.20°.

本发明的一些方案中，上述B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°，14.23±0.20°，17.16±0.20°，19.33±0.20°，20.03±0.20°，20.91±0.20°，21.46±0.20°和23.91±0.20°。In some embodiments of the present invention, the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 17.16±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20° and 23.91±0.20°.

本发明的一些方案中，上述B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°，14.23±0.20°，14.60±0.20°，17.16±0.20°，19.33±0.20°，20.03±0.20°，20.91±0.20°，21.46±0.20°，23.91±0.20°，24.49±0.20°，25.15±0.20°和27.14±0.20°。In some embodiments of the present invention, the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 14.60±0.20°, 17.16±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20°, 23.91±0.20°, 24.49±0.20°, 25.15±0.20° and 27.14±0.20°.

本发明的一些方案中，上述B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°，14.23±0.20°，14.60±0.20°，17.16±0.20°，19.33±0.20°，20.03±0.20°，20.91±0.20°，21.46±0.20°，21.90±0.20°，22.36±0.20°，23.60±0.20°，23.91±0.20°，24.49±0.20°，25.15±0.20°，27.14±0.20°和29.89±0.20°。In some embodiments of the present invention, the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 14.60±0.20°, 17.16±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20°, 21.90±0.20°, 22.36±0.20°, 23.60±0.20°, 23.91±0.20°, 24.49±0.20°, 25.15±0.20°, 27.14±0.20° and 29.89±0.20°.

本发明的一些方案中，上述B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°，9.40±0.20°，10.83±0.20°，12.39±0.20°，14.23±0.20°，14.60±0.20°，15.31±0.20°，15.99±0.20°，16.24±0.20°，17.16±0.20°，18.96±0.20°，19.33±0.20°，20.03±0.20°，20.91±0.20°，21.46±0.20°，21.90±0.20°，22.36±0.20°，23.60±0.20°，23.91±0.20°，24.49±0.20°，24.88±0.20°，25.15±0.20°，26.66±0.20°，27.14±0.20°，27.78±0.20°，28.29±0.20°，29.22±0.20°，29.44±0.20°，29.89±0.20°，31.48±0.20°，32.39±0.20°，33.22±0.20°和34.96±0.20°。In some embodiments of the present invention, the above-mentioned B crystal form has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 9.40±0.20°, 10.83±0.20°, 12.39±0.20°, 14.23±0.20°, 14.60±0.20°, 15.31±0.20°, 15.99±0.20°, 16.24±0.20°, 17.16±0.20°, 18.96±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20°. °, 21.90±0.20°, 22.36±0.20°, 23.60±0.20°, 23.91±0.20°, 24.49±0.20°, 24.88±0.20°, 25.15±0.20°, 26.66±0.20°, 27.14±0.20°, 27.78±0.20°, 28.29±0.20°, 29.22±0.20°, 29.44±0.20°, 29.89±0.20°, 31.48±0.20°, 32.39±0.20°, 33.22±0.20° and 34.96±0.20°.

本发明的一些方案中，上述B晶型的XRPD图谱解析数据基本上如表2所示。In some embodiments of the present invention, the XRPD spectrum analysis data of the above-mentioned Form B is basically as shown in Table 2.

表2化合物TM03的B晶型的XRPD图谱解析数据

Table 2 XRPD spectrum analysis data of Form B of compound TM03

本发明中，化合物TM03的B晶型，其XRPD图谱基本上如图9所示。In the present invention, the XRPD spectrum of Form B of compound TM03 is basically as shown in FIG. 9 .

本发明的一些方案中，化合物TM03的B晶型，其特征在于，具备下列任意一项特征：In some embodiments of the present invention, the B crystal form of compound TM03 is characterized by having any one of the following characteristics:

(1)其差示扫描量热曲线在187.33℃±5℃处具有吸热峰的起始点；(1) Its differential scanning calorimetry curve has an endothermic peak starting point at 187.33°C ± 5°C;

(2)其DSC图谱基本上如图10所示；(2) Its DSC spectrum is substantially as shown in FIG10 ;

(3)其热重分析曲线在30-200℃时无失重；(3) Its thermogravimetric analysis curve shows no weight loss at 30-200°C;

(4)其TGA图谱基本上如图11所示。(4) Its TGA spectrum is basically as shown in Figure 11.

本发明还提供化合物TM03的B晶型的制备方法，包括如下(a)～(c)的步骤：The present invention also provides a method for preparing the B crystal form of compound TM03, comprising the following steps (a) to (c):

(a)将化合物TM03加入甲醇中加热使其成澄清液；(a) Compound TM03 was added to methanol and heated to form a clear liquid;

(b)然后体系降温至至20～25℃，并在此温度下搅拌12小时；(b) then cooling the system to 20-25° C. and stirring at this temperature for 12 hours;

(c)过滤后干燥8～16小时。(c) After filtering, dry for 8 to 16 hours.

本发明还提供化合物TM03的D晶型，其特征在于，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：3.55±0.20°，18.57±0.20°和20.63±0.20°；
The present invention also provides a D crystal form of compound TM03, characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 3.55±0.20°, 18.57±0.20° and 20.63±0.20°;

本发明的一些方案中，上述D晶型的XRPD图谱解析数据基本上如表4所示。In some embodiments of the present invention, the XRPD spectrum analysis data of the above-mentioned D crystal form is basically as shown in Table 4.

表5化合物TM03的D晶型的XRPD图谱解析数据
Table 5 XRPD spectrum analysis data of D crystal form of compound TM03

本发明中，化合物TM03的D晶型，其XRPD图谱基本上如图13所示。In the present invention, the XRPD spectrum of the D crystal form of compound TM03 is basically as shown in FIG. 13 .

在本发明的一些方案中，上述D晶型的差示扫描量热曲线在180.18℃±5℃，327.60℃±5℃处各有一个吸热峰的起始点；在128.94℃±5℃处具有放热峰的起始点。In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned D crystal form has an endothermic peak starting point at 180.18℃±5℃ and 327.60℃±5℃ respectively; and has an exothermic peak starting point at 128.94℃±5℃.

在本发明的一些方案中，上述D晶型的DSC图谱基本上如图14所示。In some embodiments of the present invention, the DSC spectrum of the above-mentioned D crystal form is basically as shown in Figure 14.

在本发明的一些方案中，上述D晶型的热重分析曲线在165℃时失重达2.861％。In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned D crystal form shows a weight loss of 2.861% at 165°C.

在本发明的一些方案中，上述D晶型的TGA图谱基本上如图15所示。In some embodiments of the present invention, the TGA spectrum of the above-mentioned D crystal form is basically as shown in Figure 15.

本发明还提供化合物TM03的D晶型的制备方法，包括如下(a)～(c)的步骤：The present invention also provides a method for preparing the D crystal form of compound TM03, comprising the following steps (a) to (c):

(a)将化合物TM03加乙酸乙酯中，体系溶澄清； (a) Compound TM03 was added to ethyl acetate, and the system became clear;

(b)然后除去乙酸乙酯，加入正庚烷，体系在20～25℃下搅拌144小时；(b) then removing ethyl acetate, adding n-heptane, and stirring the system at 20-25° C. for 144 hours;

(c)过滤后干燥8～16小时。(c) After filtering, dry for 8 to 16 hours.

本发明还提供了上述的化合物、其立体异构体或其药学上可接受的盐在制备治疗P2X3抑制剂相关疾病的药物上的应用。The present invention also provides the use of the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt in the preparation of drugs for treating diseases related to P2X3 inhibitors.

本发明还提供了化合物TM03的晶型在制备治疗P2X3抑制剂相关疾病的药物上的应用。The present invention also provides the use of the crystalline form of compound TM03 in the preparation of drugs for treating diseases related to P2X3 inhibitors.

本发明的一些方案中，所述P2X3抑制剂相关疾病是指疼痛或慢性咳嗽。In some embodiments of the present invention, the P2X3 inhibitor-related disease refers to pain or chronic cough.

本发明的一些方案中，所述P2X3抑制剂相关疾病是指难治性咳嗽和/或COVID-19相关咳嗽。In some embodiments of the present invention, the P2X3 inhibitor-related disease refers to refractory cough and/or COVID-19-related cough.

本发明还提供了如下的合成方法：The present invention also provides the following synthesis method:

方法1：
Method 1:

方法2：
Method 2:

方法3：
Method 3:

方法4：
Method 4:

方法5：
Method 5:

方法6：
Method 6:

方法7：
Method 7:

方法8：

Method 8:

方法9：
Method 9:

方法10：
Method 10:

本发明还提供了如下的测试方法：The present invention also provides the following testing method:

测试方法1：本发明化合物的体外酶活性测试Test Method 1: In vitro enzyme activity test of the compounds of the present invention

采用33P同位素标记激酶活性测试(Reaction Biology Corp)测定IC50值来评价受试化合物对P2X3受体的抑制能力。The IC 50 value was determined using 33 P isotope-labeled kinase activity assay (Reaction Biology Corp) to evaluate the inhibitory ability of the test compounds on P2X3 receptors.

缓冲液条件：20mM羟乙基哌嗪乙硫磺酸(Hepes)(pH 7.5)，10mM氯化镁(MgCl2)，1mM乙二醇双氨乙基醚四乙酸(EGTA)，0.02％聚氧乙烯十二烷醚(Brij35)，0.02mg/ml BSA，0.1mM钒酸钠(Na3VO4)，2mM二硫苏糖醇(DTT)，1％DMSO。Buffer conditions: 20 mM Hepes (pH 7.5), 10 mM MgCl 2 , 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na vanadate (Na 3 VO 4 ), 2 mM DTT, 1% DMSO.

试验步骤：室温下，将受试化合物溶解在DMSO中配制成10mM溶液待用。将底物溶解在新配制的缓冲液中，向其中加入受测激酶并混合均匀。利用声学技术(Echo 550)将溶有受试化合物的DMSO溶液加入上述混匀的反应液中。反应液中化合物浓度为10μM，2.50μM，0.62μM，0.156μM，39.1nM，9.8nM，2.4nM，0.61nM，0.15nM，0.038nM或3μM，1μM，0.333μM，0.111μM，37.0nM，12.3nM，4.12nM，1.37nM，0.457nM，0.152nM。孵化15分钟后，加入33P-ATP(活度0.01μCi/μL)开始反应。反应在室温下进行120分钟后，将反应液点在P81离子交换滤纸(Whatman#3698-915)上。用0.75％磷酸溶液反复清洗滤纸后，测定滤纸上残留的磷酸化底物的放射性。激酶活性数据用含有受试化合物的激酶活性和空白组(仅含有DMSO)的激酶活性的比对表示，通过Prism4软件(GraphPad)进行曲线拟合得到IC50值。Test steps: At room temperature, the test compound was dissolved in DMSO to prepare a 10mM solution for use. The substrate was dissolved in a freshly prepared buffer, to which the kinase to be tested was added and mixed evenly. The DMSO solution containing the test compound was added to the above mixed reaction solution using acoustic technology (Echo 550). The concentration of the compound in the reaction solution was 10μM, 2.50μM, 0.62μM, 0.156μM, 39.1nM, 9.8nM, 2.4nM, 0.61nM, 0.15nM, 0.038nM or 3μM, 1μM, 0.333μM, 0.111μM, 37.0nM, 12.3nM, 4.12nM, 1.37nM, 0.457nM, 0.152nM. After 15 minutes of incubation, 33 P-ATP (activity 0.01 μCi/μL) was added to start the reaction. After the reaction was carried out at room temperature for 120 minutes, the reaction solution was spotted on P81 ion exchange filter paper (Whatman #3698-915). After the filter paper was repeatedly washed with 0.75% phosphoric acid solution, the radioactivity of the phosphorylated substrate remaining on the filter paper was measured. The kinase activity data were expressed as a comparison of the kinase activity of the test compound and the kinase activity of the blank group (containing only DMSO), and the IC 50 value was obtained by curve fitting using Prism4 software (GraphPad).

技术效果Technical Effects

本发明化合物对P2X3受体具有优异的抑制活性；具有较好的暴露量和生物利用度；能显著降低ATP+柠檬酸诱导豚鼠咳嗽次数，且能显著升高ATP+柠檬酸诱导豚鼠咳嗽潜伏期。 The compound of the present invention has excellent inhibitory activity on P2X3 receptors, has good exposure and bioavailability, can significantly reduce the number of coughs induced by ATP+citric acid in guinea pigs, and can significantly increase the latency of coughs induced by ATP+citric acid in guinea pigs.

附图说明BRIEF DESCRIPTION OF THE DRAWINGS

图1.化合物1的结合模式图；Figure 1. Binding mode diagram of compound 1;

图2.化合物2的结合模式图；Figure 2. Binding mode diagram of compound 2;

图3.化合物3的结合模式图；Figure 3. Binding mode diagram of compound 3;

图4.化合物4的结合模式图；Figure 4. Binding mode diagram of compound 4;

图5.化合物5的结合模式图；Figure 5. Binding mode diagram of compound 5;

图6.化合物TM03的A晶型的Cu-Kα辐射的XRPD谱图；Figure 6. XRPD spectrum of Form A of compound TM03 using Cu-Kα radiation;

图7.化合物TM03的A晶型的DSC谱图；Figure 7. DSC spectrum of Form A of compound TM03;

图8.化合物TM03的A晶型的TGA谱图；Figure 8. TGA spectrum of Form A of compound TM03;

图9.化合物TM03的B晶型的Cu-Kα辐射的XRPD谱图；Figure 9. XRPD spectrum of Form B of compound TM03 using Cu-Kα radiation;

图10.化合物TM03的B晶型的DSC谱图；Figure 10. DSC spectrum of Form B of compound TM03;

图11.化合物TM03的B晶型的TGA谱图；Figure 11. TGA spectrum of Form B of compound TM03;

图12.化合物TM03的B晶型的DVS谱图；Figure 12. DVS spectrum of Form B of compound TM03;

图13.化合物TM03的D晶型的Cu-Kα辐射的XRPD谱图；Figure 13. XRPD spectrum of the D-form of compound TM03 using Cu-Kα radiation;

图14.化合物TM03的D晶型的DSC谱图；Figure 14. DSC spectrum of Form D of compound TM03;

图15.化合物TM03的D晶型的TGA谱图。Figure 15. TGA spectrum of Form D of compound TM03.

定义和说明Definition and Description

除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其他问题或并发症，与合理的利益/风险比相称。Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of special definition, but should be understood according to the ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient. The term "pharmaceutically acceptable" used here refers to those compounds, materials, compositions and/or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, and are commensurate with a reasonable benefit/risk ratio.

术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物的中性形式接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机氨或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物的中性形式接触的方式获得酸加成盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。The term "pharmaceutically acceptable salt" refers to salts of compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base or acid addition salts.

本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention.

除非另有说明，术语“对映异构体”或者“旋光异构体”是指互为镜像关系的立体异构体。Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

除非另有说明，术语“顺反异构体”或者“几何异构体”系由因双键或者成环碳原子单键不能自由旋转而引起。Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability of a ring to rotate freely about double bonds or single bonds of ring carbon atoms.

除非另有说明，术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

除非另有说明，“(+)”表示右旋，“(-)”表示左旋，“(±)”表示外消旋。Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

除非另有说明，用楔形实线键和楔形虚线键表示一个立体中心的绝对构型，用直形实 线键和直形虚线键表示立体中心的相对构型，用波浪线表示楔形实线键或楔形虚线键或用波浪线表示直形实线键和直形虚线键 Unless otherwise specified, the key is a solid wedge. and dotted wedge key To express the absolute configuration of a stereocenter, use a straight Line Key and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Denotes a solid wedge bond or dotted wedge key Or use a wavy line Represents a straight solid bond and straight dashed key

除非另有说明，当化合物中存在双键结构，如碳碳双键、碳氮双键和氮氮双键，且双键上的各个原子均连接有两个不同的取代基时(包含氮原子的双键中，氮原子上的一对孤对电子视为其连接的一个取代基)，如果该化合物中双键上的原子与其取代基之间用波浪线连接，则表示该化合物的(Z)型异构体、(E)型异构体或两种异构体的混合物。例如下式(A)表示该化合物以式(A-1)或式(A-2)的单一异构体形式存在或以式(A-1)和式(A-2)两种异构体的混合物形式存在；下式(B)表示该化合物以式(B-1)或式(B-2)的单一异构体形式存在或以式(B-1)和式(B-2)两种异构体的混合物形式存在。下式(C)表示该化合物以式(C-1)或式(C-2)的单一异构体形式存在或以式(C-1)和式(C-2)两种异构体的混合物形式存在。
Unless otherwise specified, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (in a double bond containing a nitrogen atom, a lone pair of electrons on the nitrogen atom is regarded as a substituent connected to it), if a wavy line is used between the atom on the double bond and its substituent in the compound, If connected, it means the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound. For example, the following formula (A) means that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2) or in the form of a mixture of two isomers of formula (A-1) and formula (A-2); the following formula (B) means that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2) or in the form of a mixture of two isomers of formula (B-1) and formula (B-2). The following formula (C) means that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2) or in the form of a mixture of two isomers of formula (C-1) and formula (C-2).

除非另有说明，术语“互变异构体”或“互变异构体形式”是指在室温下，不同官能团异构体处于动态平衡，并能很快的相互转化。若互变异构体是可能的(如在溶液中)，则可以达到互变异构体的化学平衡。例如，质子互变异构体(proton tautomer)(也称质子转移互变异构体(prototropic tautomer))包括通过质子迁移来进行的互相转化，如酮-烯醇异构化和亚胺-烯胺异构化。价键异构体(valencetautomer)包括一些成键电子的重组来进行的相互转化。其中酮-烯醇互变异构化的具体实例是戊烷-2，4-二酮与4-羟基戊-3-烯-2-酮两个互变异构体之间的互变，再如与两个互变异构体之间的互变。Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and can rapidly convert into each other. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence isomers include interconversions via reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one. For example and Interconversion between two tautomers.

除非另有说明，术语“富含一种异构体”、“异构体富集”、“富含一种对映体”或者“对映体富集”指其中一种异构体或对映体的含量小于100％，并且，该异构体或对映体的含量大于等于60％，或者大于等于70％，或者大于等于80％，或者大于等于90％，或者大于等于95％，或者大于等于96％，或者大于等于97％，或者大于等于98％，或者大于等于99％，或者大于等于99.5％，或者大于等于99.6％，或者大于等于99.7％，或者大于等于99.8％，或者大于等于99.9％。Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

除非另有说明，术语“异构体过量”或“对映体过量”指两种异构体或两种对映体相对百分数之间的差值。例如，其中一种异构体或对映体的含量为90％，另一种异构体或对映体的含量为10％，则异构体或对映体过量(ee值)为80％。Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备， 其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary. Wherein the resulting diastereomeric mixture is separated and the auxiliary groups are cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art and then the pure enantiomers are recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamates are generated from amines).

本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚(3H)，碘-125(125I)或C-14(14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound. For example, the compound may be labeled with a radioactive isotope, such as tritium ( 3H ), iodine-125 ( 125I ) or C-14 ( 14C ). For another example, deuterated drugs may be formed by replacing hydrogen with heavy hydrogen. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing therapeutic effects, and extending the biological half-life of drugs. All isotopic composition changes of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.

“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。"Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，可以包括重氢和氢的变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧(即＝O)时，意味着两个氢原子被取代。氧取代不会发生在芳香基上。术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or not substituted, and unless otherwise specified, the type and number of the substituent can be arbitrary on the basis of chemical achievable.

当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice. In addition, combinations of substituents and/or variants thereof are permitted only if such combinations result in stable compounds.

当一个连接基团的数量为0时，比如-(CRR)0-，表示该连接基团为单键。When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.

当其中一个变量选自单键时，表示其连接的两个基团直接相连，比如A-L-Z中L代表单键时表示该结构实际上是A-Z。When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.

当一个取代基为空缺时，表示该取代基是不存在的，比如A-X中X为空缺时表示该结构实际上是A。当所列举的取代基中没有指明其通过哪一个原子连接到被取代的基团上时，这种取代基可以通过其任何原子相键合，例如，吡啶基作为取代基可以通过吡啶环上任意一个碳原子连接到被取代的基团上。当所列举的连接基团没有指明其连接方向，其连接方向是任意的，例如，中连接基团L为-M-W-，此时-M-W-既可以按与从左往右的读取顺序相同的方向连接环A和环B构成也可以按照与从左往右的读取顺序相反的方向连接环A和环B构成所述连接基团、取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A. When the listed substituent does not specify which atom it is connected to the substituted group through, the substituent can be bonded through any atom of it. For example, pyridyl as a substituent can be connected to the substituted group through any carbon atom on the pyridine ring. When the listed connecting group does not specify its connection direction, its connection direction is arbitrary. For example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of the reading order from left to right to form Combinations of linkers, substituents, and/or variations thereof are permissible only if such combinations result in stable compounds.

除非另有规定，术语“C1-4烷基”用于表示直链或支链的由1至4个碳原子组成的饱和碳氢基团。所述C1-4烷基包括C1-2、C1-3和C2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C1-4烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)等。Unless otherwise specified, the term "C 1-4 alkyl" is used to represent a straight or branched saturated hydrocarbon group consisting of 1 to 4 carbon atoms. The C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc.; they can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of C 1-4 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), etc.

除非另有规定，术语“C1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C1-3烷基包括C1-2和C2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C1- 3烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)等。 Unless otherwise specified, the term "C 1-3 alkyl" is used to represent a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of C 1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

除非另有规定，术语“C1-4烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至4个碳原子的烷基基团。所述C1-4烷氧基包括C1-3、C1-2、C2-4、C4和C3烷氧基等。C1-4烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)、丁氧基(包括n-丁氧基、异丁氧基、s-丁氧基和t-丁氧基)等。Unless otherwise specified, the term "C 1-4 alkoxy" refers to those alkyl groups containing 1 to 4 carbon atoms connected to the rest of the molecule through an oxygen atom. The C 1-4 alkoxy includes C 1-3 , C 1-2 , C 2-4 , C 4 and C 3 alkoxy, etc. Examples of C 1-4 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), etc.

除非另有规定，术语“C1-3烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至3个碳原子的烷基基团。所述C1-3烷氧基包括C1-2、C2-3、C3和C2烷氧基等。C1-3烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)等。Unless otherwise specified, the term "C 1-3 alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms connected to the rest of the molecule through an oxygen atom. The C 1-3 alkoxy includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy, etc. Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

除非另有规定，术语“C1-3卤代烷基”表示包含1至3个碳原子的单卤代烷基和多卤代烷基。所述C1-3卤代烷基包括C1-2、C2-3、C3、C2和C1卤代烷基等。C1-3卤代烷基的实例包括但不限于三氟甲基、三氯甲基、2，2，2-三氟乙基、五氟乙基、五氯乙基、3-溴丙基等。Unless otherwise specified, the term "C 1-3 haloalkyl" refers to monohaloalkyl and polyhaloalkyl groups containing 1 to 3 carbon atoms. The C 1-3 haloalkyl group includes C 1-2 , C 2-3 , C 3 , C 2 and C 1 haloalkyl groups, etc. Examples of C 1-3 haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, pentachloroethyl, 3-bromopropyl, etc.

除非另有规定，术语“C1-4烷氨基”表示通过氨基连接到分子的其余部分的那些包含1至4个碳原子的烷基基团。所述C1-4烷氨基包括C1-3、C1-2、C2-4、C4、C3和C2烷氨基等。C1-4烷氨基的实例包括但不限于-NHCH3、-N(CH3)2、-NHCH2CH3、-N(CH3)CH2CH3、-N(CH2CH3)(CH2CH3)、-NHCH2CH2CH3、-NHCH2(CH3)2、-NHCH2CH2CH2CH3等。Unless otherwise specified, the term "C 1-4 alkylamino" refers to those alkyl groups containing 1 to 4 carbon atoms attached to the rest of the molecule through an amino group. The C 1-4 alkylamino group includes C 1-3 , C 1-2 , C 2-4 , C 4 , C 3 and C 2 alkylamino groups, etc. Examples of C 1-4 alkylamino groups include, but are not limited to, -NHCH 3 , -N(CH 3 ) 2 , -NHCH 2 CH 3 , -N(CH 3 )CH 2 CH 3 , -N(CH 2 CH 3 )(CH 2 CH 3 ), -NHCH 2 CH 2 CH 3 , -NHCH 2 (CH 3 ) 2 , -NHCH 2 CH 2 CH 2 CH 3 , and the like.

除非另有规定，本发明术语“5-6元杂芳环”和“5-6元杂芳基”可以互换使用，术语“5-6元杂芳基”表示由5至6个环原子组成的具有共轭π电子体系的单环基团，其1、2、3或4个环原子为独立选自O、S和N的杂原子，其余为碳原子。其中氮原子任选地被季铵化，氮和硫杂原子可任选被氧化(即NO和S(O)p，p是1或2)。5-6元杂芳基可通过杂原子或碳原子连接到分子的其余部分。所述5-6元杂芳基包括5元和6元杂芳基。所述5-6元杂芳基的实例包括但不限于吡咯基(包括N-吡咯基、2-吡咯基和3-吡咯基等)、吡唑基(包括2-吡唑基和3-吡唑基等)、咪唑基(包括N-咪唑基、2-咪唑基、4-咪唑基和5-咪唑基等)、噁唑基(包括2-噁唑基、4-噁唑基和5-噁唑基等)、三唑基(1H-1，2，3-三唑基、2H-1，2，3-三唑基、1H-1，2，4-三唑基和4H-1，2，4-三唑基等)、四唑基、异噁唑基(3-异噁唑基、4-异噁唑基和5-异噁唑基等)、噻唑基(包括2-噻唑基、4-噻唑基和5-噻唑基等)、呋喃基(包括2-呋喃基和3-呋喃基等)、噻吩基(包括2-噻吩基和3-噻吩基等)、吡啶基(包括2-吡啶基、3-吡啶基和4-吡啶基等)、吡嗪基或嘧啶基(包括2-嘧啶基和4-嘧啶基等)。Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" of the present invention can be used interchangeably, and the term "5-6 membered heteroaryl" means a monocyclic group consisting of 5 to 6 ring atoms with a conjugated π electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. Wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2). The 5-6 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl) and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

除非另有规定，术语“卤代素”或“卤素”本身或作为另一取代基的一部分表示氟、氯、溴或碘原子。[0043] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention.

本发明的中间体化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。The intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention.

本发明具体实施方式的化学反应是在合适的溶剂中完成的，所述的溶剂须适合于本发明的化学变化及其所需的试剂和物料。为了获得本发明的化合物，有时需要本领域技术人员在已有实施方式的基础上对合成步骤或者反应流程进行修改或选择。The chemical reactions of the specific embodiments of the present invention are carried out in a suitable solvent, which must be suitable for the chemical changes of the present invention and the reagents and materials required. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments.

本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：扫描，收集相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。The structure of the compound of the present invention can be confirmed by conventional methods known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data of the cultured single crystal using a Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

本发明所使用的溶剂可经市售获得。本发明采用下述缩略词：DMSO代表二甲亚砜；EtOH代表乙醇；ACN代表乙腈；hr代表小时；EA代表乙酸乙酯；MTBE代表甲基叔丁基醚；THF代表四氢呋喃；PE代表石油醚。The solvent used in the present invention can be obtained from commercial sources. The present invention uses the following abbreviations: DMSO represents dimethyl sulfoxide; EtOH represents ethanol; ACN represents acetonitrile; hr represents hour; EA represents ethyl acetate; MTBE represents methyl tert-butyl ether; THF represents tetrahydrofuran; and PE represents petroleum ether.

化合物依据本领域常规命名原则或者使用软件命名，市售化合物采用供应商目录名称。Compounds are named according to the conventional nomenclature in the art or using The software names were used, and commercially available compounds were named using the supplier's catalog names.

本发明X射线粉末衍射(X-ray powder diffractometer，XRPD)方法，测试参数见表5。The X-ray powder diffractometer (XRPD) method of the present invention, the test parameters are shown in Table 5.

表5 XRPD测试参数
Table 5 XRPD test parameters

本发明差热分析(Differential Scanning Calorimeter，DSC)方法，测试参数见表6。The differential scanning calorimeter (DSC) method of the present invention and the test parameters are shown in Table 6.

表6 DSC测试参数
Table 6 DSC test parameters

本发明热重分析(Thermal Gravimetric Analyzer，TGA)方法，测试参数见表7。Thermogravimetric analysis (TGA) method of the present invention, test parameters are shown in Table 7.

表7 TGA测试参数
Table 7 TGA test parameters

本发明动态气体吸附分析(Dynamic Vapor Sorption，DVS)方法，测试参数见表8。The dynamic vapor sorption analysis (DVS) method of the present invention, the test parameters are shown in Table 8.

表8 DVS测试参数

Table 8 DVS test parameters

引湿性评价分类如下：

注：ΔW％表示受试品在25±1℃和80±2％RH下的吸湿增重。The moisture absorption evaluation is classified as follows:

Note: ΔW% indicates the weight gain of the test sample at 25±1℃ and 80±2%RH.

具体实施方式DETAILED DESCRIPTION

下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。The present invention is described in detail below by way of examples, but it is not intended to impose any adverse limitations on the present invention. The present invention has been described in detail herein, and specific embodiments thereof are also disclosed therein. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

计算例1.本发明化合物与hP2X3离子通道的结合模式预测
Calculation Example 1. Prediction of the binding mode of the compound of the present invention to the hP2X3 ion channel

本次结合模式预测使用hP2X3共晶结构(PDB ID代码：5SVR)作为对接模板。为了制备蛋白质，使用Maestro(版本2021-2)[1]的蛋白质制备向导模块添加氢原子，并使用OPLS4力场。并对共晶结构进行了氢键优化，去除共晶小分子以外的水分子，以及整体能量优化。首先对盐野义公司公开的分子S-600918进行对接确定其结合模式。用Glide[2]Receptor Grid Generation模块以共晶结构已知的hP2X3拮抗剂A-317491的质心生成的对接网格，并用Ligand Docking模块寻找出S-600918的最佳结合模式。该结合模式下S-600918的药效团以及关键蛋白-配体相互作用与A-317491相似，证明了对接模型的可靠性。对于配体的制备：使用LigPrep[3]将新设计的分子生成了3D结构，并进行了能量最小化。基于这个对接模型，对新设计的分子用Glide[2]中的SP对接模式进行对接。化合物1～5的结合模式见图1-5。The hP2X3 cocrystal structure (PDB ID code: 5SVR) was used as the docking template for this binding mode prediction. To prepare the protein, Maestro ( The protein preparation wizard module of version 2021-2) [1] added hydrogen atoms and used the OPLS4 force field. The cocrystal structure was hydrogen bond optimized and the cocrystal small molecules were removed. The water molecules other than water molecules and the overall energy optimization were first docked to determine the binding mode of the molecule S-600918 published by Shionogi. The Glide [2] Receptor Grid Generation module was used to generate the center of mass of the hP2X3 antagonist A-317491 with a known cocrystal structure. The docking grid was used and the Ligand Docking module was used to find the optimal binding mode of S-600918. The pharmacophore and key protein-ligand interactions of S-600918 in this binding mode are similar to those of A-317491, which proves the reliability of the docking model. For the preparation of ligands: the 3D structure of the newly designed molecules was generated using LigPrep [3] and energy minimized. Based on this docking model, the newly designed molecules were docked using the SP docking mode in Glide [2] . The binding modes of compounds 1 to 5 are shown in Figures 1-5.

[1]Maestro，LLC，New York，NY，2021.[1] Maestro, LLC, New York, NY, 2021.

[2]Glide，LLC，New York，NY，2021. [2] Glide, LLC, New York, NY, 2021.

[3]LigPrep，LLC，New York，NY，2021.[3] LigPrep, LLC, New York, NY, 2021.

结论：本发明化合物与hP2X3离子通道有较好的结合，很好的复现了阳参分子S-600918的结合模式：本系列分子跟Arg281、Lys299、Lys63形成盐桥，跟Asn279、Ser275形成氢键。另外2-苯氧基吡啶片段插入疏水口袋跟Phe174形成π-π堆积，氯苯片段和Lys65形成阳离子-pi作用，和Thr172有静电相互作用。新设计分子对接打分接近或优于阳参分子，与hP2X3离子通道的结合将有效抑制气道的感觉神经元过度兴奋，缓解咳嗽症状。Conclusion: The compounds of the present invention have good binding with hP2X3 ion channels, and well reproduce the binding mode of Yangshen molecule S-600918: this series of molecules form salt bridges with Arg281, Lys299, and Lys63, and form hydrogen bonds with Asn279 and Ser275. In addition, the 2-phenoxypyridine fragment inserts into the hydrophobic pocket to form π-π stacking with Phe174, the chlorobenzene fragment forms a cation-pi interaction with Lys65, and has an electrostatic interaction with Thr172. The docking score of the newly designed molecule is close to or better than that of the Yangshen molecule. The binding with the hP2X3 ion channel will effectively inhibit the overexcitation of airway sensory neurons and relieve cough symptoms.

实施例1
Example 1

步骤1：化合物TM01_2的合成Step 1: Synthesis of compound TM01_2

将化合物TM01_1(5g，34.21mmol)加到乙醇(50mL)中，然后滴入浓硫酸(2.79mL，浓度：98％)，80℃搅拌12hr。反应完毕，将体系降至室温，减压浓缩除去溶剂。浓缩物加水(30mL)，然后用乙酸乙酯(20mL×2)萃取，合并有机相，有机相用饱和碳酸氢钠溶液(20mL×2)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂，得到化合物TM01_2，直接用于下一步。MS m/z：203.1[M+1]+。1H NMR(400MHz，CDCl3)：δ＝4.07-4.18(m，4H)，2.42-2.52(m，1H)，2.29-2.36(m，2H)，1.89-2.02(m，1H)，1.71-1.83(m，1H)，1.22-1.30(m，6H)，1.14-1.20ppm(m，3H).Compound TM01_1 (5 g, 34.21 mmol) was added to ethanol (50 mL), and then concentrated sulfuric acid (2.79 mL, concentration: 98%) was added dropwise, and stirred at 80°C for 12 hours. After the reaction was completed, the system was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The concentrate was added with water (30 mL), and then extracted with ethyl acetate (20 mL×2). The organic phases were combined, washed with saturated sodium bicarbonate solution (20 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent to obtain compound TM01_2, which was directly used in the next step. MS m/z: 203.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ = 4.07-4.18 (m, 4H), 2.42-2.52 (m, 1H), 2.29-2.36 (m, 2H), 1.89-2.02 (m, 1H), 1.71-1.83 (m, 1H), 1.22-1.30 (m, 6H), 1.14-1.2 0ppm(m,3H).

步骤2：化合物TM01_3的合成Step 2: Synthesis of compound TM01_3

向四氢呋喃(40mL)中加入叔丁醇钾(1M四氢呋喃溶液，49.44mL)，将体系降至0℃，然后向体系中加入溶于四氢呋喃(10mL)的化合物TM01_2(5g，24.72mmol)和甲酸乙酯(2.98mL)溶液，滴加过程控制温度不超过10℃，然后将温度升至20℃-25℃搅拌2hr。原料反应完毕，向反应液中加入水(25mL)，然后加入乙酸乙酯(25mL)，分液，向水相中加入乙酸乙酯(25mL)，合并两次的有机相，加入氢氧化钠(0.5g，12.36mmol)的水溶液(12mL)，分液，合并所有水相得到化合物TM01_3的水溶液，直接用于下一步。MS m/z：231.2[M+1]+。Potassium tert-butoxide (1M tetrahydrofuran solution, 49.44 mL) was added to tetrahydrofuran (40 mL), the system was cooled to 0°C, and then compound TM01_2 (5 g, 24.72 mmol) and ethyl formate (2.98 mL) solution dissolved in tetrahydrofuran (10 mL) were added to the system, the temperature was controlled not to exceed 10°C during the dropwise addition, and then the temperature was raised to 20°C-25°C and stirred for 2 hours. After the reaction of the raw materials was completed, water (25 mL) was added to the reaction solution, and then ethyl acetate (25 mL) was added, and the liquid was separated. Ethyl acetate (25 mL) was added to the aqueous phase, and the organic phases of the two times were combined, and an aqueous solution (12 mL) of sodium hydroxide (0.5 g, 12.36 mmol) was added, and the liquid was separated. All aqueous phases were combined to obtain an aqueous solution of compound TM01_3, which was directly used in the next step. MS m/z: 231.2 [M+1] + .

步骤3：化合物TM01_4的合成Step 3: Synthesis of compound TM01_4

将S-乙基异硫脲氢溴酸盐(4.21g，22.73mmol)加到上一步化合物TM01_3的水溶液中，40℃搅拌12hr。将反应体系降至室温，加入乙酸乙酯(30mL×3)萃取，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝1∶1)，得到化合物TM01_4。MS m/z：271.3[M+1]+。1H NMR(400MHz，CDCl3)：δ＝11.63-11.78(m，1H)，7.71(s，1H)，4.08-4.15(m，2H)，3.14-3.22(m，2H)，2.87-2.97(m，1H)，2.70-2.78(m，1H)，2.56(dd，J＝13.8，6.4Hz，1H)，1.39(t，J＝7.4Hz，3H)，1.19-1.25ppm(m，6H)。Add S-ethylisothiourea hydrobromide (4.21 g, 22.73 mmol) to the aqueous solution of compound TM01_3 in the previous step and stir at 40°C for 12 hours. Cool the reaction system to room temperature, add ethyl acetate (30 mL×3) for extraction, combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to remove the solvent. The crude product is separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:1) to obtain compound TM01_4. MS m/z: 271.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ = 11.63-11.78 (m, 1H), 7.71 (s, 1H), 4.08-4.15 (m, 2H), 3.14-3.22 (m, 2H), 2.87-2.97 (m, 1H), 2.70-2.78 (m, 1H), 2.56 (dd, J＝ 13.8, 6.4Hz, 1H), 1.39 (t, J=7.4Hz, 3H), 1.19-1.25ppm (m, 6H).

步骤4：化合物TM01_5的合成Step 4: Synthesis of compound TM01_5

将化合物TM01_4(100mg，369.89μmol)加到二氯甲烷(1mL)中，加入二异丙基乙胺(161.07μL)，然后加入对氯苄溴(76.01mg，369.89μmol)，25℃搅拌12hr。压浓缩，粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝10∶1)，得到化合物TM01_5。MS m/z：395.2[M+1]+。1H NMR(400MHz，CDCl3)δ＝7.36-7.38(d，J＝8.41Hz，2H)，7.10-7.18(m，3H)，4.95-4.98(m，2H)，3.90-3.91(br d，J＝2.26Hz，2H)，3.20-3.34(m，2H)，2.89-3.00(m，1H)，2.56-2.57(d，J＝7.40Hz，2H)，1.32-1.39(m，3H)，1.13-1.23(m，6H). Compound TM01_4 (100 mg, 369.89 μmol) was added to dichloromethane (1 mL), and diisopropylethylamine (161.07 μL) was added, followed by p-chlorobenzyl bromide (76.01 mg, 369.89 μmol), and stirred at 25°C for 12 hr. The mixture was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 10:1) to obtain compound TM01_5. MS m/z: 395.2 [M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 7.36-7.38 (d, J = 8.41Hz, 2H), 7.10-7.18 (m, 3H), 4.95-4.98 (m, 2H), 3.90-3.91 (br d, J = 2.26Hz, 2H), 3.20-3.34 (m, 2H), 2.89-3. 00 (m, 1H), 2.56-2.57 (d, J=7.40Hz, 2H), 1.32-1.39 (m, 3H), 1.13-1.23 (m, 6H).

步骤5：化合物TM01_6的合成Step 5: Synthesis of compound TM01_6

将化合物TM01_5(100mg，253.22μmol)加到叔丁醇(1mL)中，然后加入化合物TM01_5A(70.73mg，379.83μmol)和乙酸(215.11μL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(5mL×3)萃取，合并有机相，用饱和食盐水(10mL×3)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝10∶1)，得到化合物TM01_6。MS m/z：519.3[M+1]+。1H NMR(400MHz，CDCl3)δ＝8.18-8.20(m，1H)7.68-7.74(m，1H)7.36-7.37(m，4H)7.12-7.17(m，3H)6.92-7.04(m，4H)5.01-5.25(m，2H)3.95-4.15(m，2H)2.68-2.83(m，1H)2.44-2.54(m，2H)1.18-1.23(m，6H).Compound TM01_5 (100 mg, 253.22 μmol) was added to tert-butyl alcohol (1 mL), followed by compound TM01_5A (70.73 mg, 379.83 μmol) and acetic acid (215.11 μL), stirred at 115°C for 12 hr, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (5 mL×3), the organic phases were combined, washed with saturated brine (10 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=10:1) to obtain compound TM01_6. MS m/z: 519.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 8.18-8.20 (m, 1H) 7.68-7.74 (m, 1H) 7.36-7.37 (m, 4H) 7.12-7.17 (m, 3H) 6.92-7.04 (m, 4H) 5.01-5.25 (m, 2H) 3.95-4.15 (m, 2H )2.68-2.83(m,1H)2.44-2.54(m,2H)1.18-1.23(m,6H).

步骤6：化合物TM01的合成Step 6: Synthesis of compound TM01

向化合物TM01_6(80mg，154.15μmol)中加入甲醇(0.6mL)，四氢呋喃(0.6mL)和水(0.2mL)，然后加入一水合氢氧化锂(25.87mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3-4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：ACN％＝30％-60％，8.0min)纯化得到化合物TM01。MS m/z：491.3[M+1]+。1H NMR(400MHz，DMSO-d6)δ＝8.15-8.16(dd，J＝4.83，1.44Hz，1H)8.09-8.10(br s，1H)7.84-7.90(m，1H)7.47-7.52(m，2H)7.40-7.46(m，2H)7.33-7.40(m，2H)7.12-7.19(m，3H)7.03-7.08(m，1H)5.50(br s，2H)2.60-2.72(m，2H)2.29-2.40(m，1H)1.03-1.11(m，3H).Methanol (0.6 mL), tetrahydrofuran (0.6 mL) and water (0.2 mL) were added to compound TM01_6 (80 mg, 154.15 μmol), and then lithium hydroxide monohydrate (25.87 mg) was added and stirred at 25°C for 12 hr. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2 M), and ethyl acetate (5 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM01 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30 mm*3 μm; flowability: [water (0.04% HCl)-ACN]; gradient: ACN%=30%-60%, 8.0 min). MS m/z: 491.3[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δ = 8.15-8.16 (dd, J = 4.83, 1.44Hz, 1H) 8.09-8.10 (br s, 1H) 7.84-7.90 (m, 1H) 7.47-7.52 (m, 2H) 7.40-7.46 (m, 2H) 7.33-7.40 (m, 2 H)7.12-7.19(m,3H)7.03-7.08(m,1H)5.50(br s,2H)2.60-2.72(m,2H)2.29-2.40(m,1H)1.03-1.11(m,3H).

实施例2
Example 2

步骤1：化合物TM02和TM03的合成Step 1: Synthesis of compounds TM02 and TM03

对TM01(100mg，203.69μmol)进行手性拆分，通过制备SFC分离(柱型：DAICEL CHIRALPAK IG(250mm*30mm，10μm)；流动相：[CO2-EtOH]；EtOH％：40％；分离时间：15mins)分离得到化合物TM02(保留时间：7.145min，ee值为100％)和化合物TM03(保留时间：8.106min，ee值为97.14％)。TM01 (100 mg, 203.69 μmol) was subjected to chiral separation and separated by preparative SFC separation (column type: DAICEL CHIRALPAK IG (250 mm*30 mm, 10 μm); mobile phase: [CO 2 -EtOH]; EtOH%: 40%; separation time: 15 mins) to give compound TM02 (retention time: 7.145 min, ee value: 100%) and compound TM03 (retention time: 8.106 min, ee value: 97.14%).

SFC分析方法：Column：Chiralpak IG-3，100×4.6mm，I.D.，3μm，流动相：A：Hexane B：EtOH(0.1％TFA，v/v)，梯度：A∶B＝80∶20流速：1mL/min，柱温：30℃，分离时间：15mins。SFC analysis method: Column: Chiralpak IG-3, 100×4.6mm, I.D., 3μm, mobile phase: A: Hexane B: EtOH (0.1% TFA, v/v), gradient: A: B=80:20 flow rate: 1mL/min, column temperature: 30℃, separation time: 15mins.

化合物TM02：MS m/z：491.1[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 11.93-12.32(m，1H)8.55-8.76(m，1H)8.10-8.23(m，1H)7.77-7.87(m，1H)7.52(br s，1H)7.37-7.48(m，4H)7.28(br d，J＝6.78Hz，1H)6.97-7.12(m，4H)5.12-5.34(m，2H)2.61-2.79(m，2H)2.07-2.23(m，1H)1.02(br d，J＝6.02Hz，3H).Compound TM02: MS m/z: 491.1[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δppm 11.93-12.32 (m, 1H) 8.55-8.76 (m, 1H) 8.10-8.23 (m, 1H) 7.77-7.87 (m, 1H) 7.52 (br s, 1H) 7.37-7.48 (m, 4H) 7.28 (br d, J = 6.78Hz, 1H) 6.97-7.12 (m, 4H) 5.12-5.34 (m , 2H) 2.61-2.79 (m, 2H) 2.07-2.23 (m, 1H) 1.02 (br d, J=6.02Hz, 3H).

化合物TM03：MS m/z：491.1[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 11.98-12.27(m，1H)8.55-8.70(m，1H)8.06-8.22(m，1H)7.75-7.87(m，1H)7.52(s，1H)7.37-7.49(m，4H)7.28(br d，J＝7.78Hz，1H)6.98-7.12(m，4H)5.10-5.37(m，2H)2.53-2.58(m，2H)2.13-2.24(m，1H)1.02(br d，J＝6.65Hz，3H).Compound TM03: MS m/z: 491.1[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δppm 11.98-12.27 (m, 1H) 8.55-8.70 (m, 1H) 8.06-8.22 (m, 1H) 7.75-7.87 (m, 1H) 7.52 (s, 1H) 7.37-7.49 (m, 4H) 7.28 (br d, J = 7.78Hz, 1H) 6.98-7.12 (m, 4H) 5.10-5.37 (m, 2H) 2.53-2.58 (m, 2H) 2.13-2.24 (m, 1H) 1.02 (br d, J=6.65Hz, 3H).

实施例3
Example 3

步骤1：化合物TM04_2的合成Step 1: Synthesis of compound TM04_2

将化合物TM01_5((540mg，1.37mmol)加到叔丁醇(6mL)中，然后加入化合物TM04_1(418.83mg， 2.05mmol)和乙酸(1.16mL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(10mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和食盐水(20mL×3)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM04_2。MSm/z：537.1[M+1]+。1H NMR(400MHz，CDCl3)：δ＝7.74-7.78(m，1H)，7.51-7.57(m，1H)，7.32-7.39(m，4H)，7.11-7.15(m，2H)，6.88(d，J＝8.7Hz，2H)，6.73-6.75(m，1H)，6.59-6.63(m，1H)，4.93-5.11(m，2H)，3.96-4.09(m，2H)，2.71-2.82(m，1H)，2.37-2.52(m，2H)，1.15-1.23ppm(m，6H).Compound TM01_5 (540 mg, 1.37 mmol) was added to tert-butyl alcohol (6 mL), and then compound TM04_1 (418.83 mg, 2.05mmol) and acetic acid (1.16mL), stirred at 115℃ for 12hr, cooled to room temperature, diluted with saturated sodium bicarbonate solution (10mL), extracted with ethyl acetate (10mL×3), combined organic phases, washed with saturated brine (20mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM04_2. MSm/z: 537.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ = 7.74-7.78 (m, 1H), 7.51-7.57 (m, 1H), 7.32-7.39 (m, 4H), 7.11-7.15 (m, 2H), 6.88 (d, J = 8.7Hz, 2H), 6.73-6.75 (m, 1H), 6.59- 6.63 (m, 1H), 4.93-5.11 (m, 2H), 3.96-4.09 (m, 2H), 2.71-2.82 (m, 1H), 2.37-2.52 (m, 2H), 1.15-1.23ppm (m, 6H).

步骤2：化合物TM04的合成Step 2: Synthesis of compound TM04

将化合物TM04_2(190mg，353.83μmol)溶于甲醇(1.8mL)，四氢呋喃(1.8mL)和水(0.6mL)，然后加入一水合氢氧化锂(59.39mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：ACN％＝30％-62％，8.0min)纯化得到化合物TM04。MS m/z：509.4[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 8.01(q，J＝8.13Hz，1H)7.86(br s，1H)7.48(br d，J＝8.13Hz，2H)7.23-7.43(m，4H)7.15(br d，J＝8.13Hz，2H)6.90(br dd，J＝17.13，7.50Hz，2H)5.34(br s，2H)2.63-2.70(m，1H)2.58(br dd，J＝13.01，6.50Hz，1H)2.27(br dd，J＝13.20，7.19Hz，1H)1.05(br d，J＝6.75Hz，3H)Compound TM04_2 (190 mg, 353.83 μmol) was dissolved in methanol (1.8 mL), tetrahydrofuran (1.8 mL) and water (0.6 mL), then lithium hydroxide monohydrate (59.39 mg) was added and stirred at 25°C for 12 hr. After the reaction was completed, pH was adjusted to 3-4 with dilute hydrochloric acid (2 M), and ethyl acetate (5 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM04 was obtained by purification by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; flowability: [water (0.04% HCl)-ACN]; gradient: ACN%=30%-62%, 8.0 min). MS m/z: 509.4[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δppm 8.01 (q, J=8.13Hz, 1H) 7.86 (br s, 1H) 7.48 (br d, J=8.13Hz, 2H) 7.23-7.43 (m, 4H) 7.15 (br d, J=8.13Hz, 2 H) 6.90 (br dd, J = 17.13, 7.50Hz, 2H) 5.34 (br s, 2H) 2.63-2.70 (m, 1H) 2.58 (br dd, J = 13.01, 6.50Hz, 1H) 2.27 (br dd, J = 13.20, 7.19Hz, 1H) 1.05 (br d, J = 6.75 Hz, 3H)

实施例4
Example 4

步骤1：化合物TM05_2的合成Step 1: Synthesis of compound TM05_2

将化合物TM01_5(540mg，1.37mmol)加到叔丁醇(6mL)中，然后加入化合物TM05_1(418.83mg，2.05mmol)和乙酸(1.16mL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(10mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和食盐水(20mL×3)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM05_2。Compound TM01_5 (540 mg, 1.37 mmol) was added to tert-butyl alcohol (6 mL), followed by compound TM05_1 (418.83 mg, 2.05 mmol) and acetic acid (1.16 mL), stirred at 115°C for 12 hr, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (10 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM05_2.

MS m/z：537.4[M+1]+.1H NMR(400MHz，CDCl3)：δ＝8.04(d，J＝3.0Hz，1H)，7.41-7.48(m，1H)，7.31-7.38(m，4H)，7.07-7.12(m，3H)，6.92(d，J＝3.1Hz，1H)，6.85(d，J＝8.7Hz，2H)，4.89-5.07(m，2H)，3.95-4.08(m，2H)，2.69-2.82(m，1H)，2.37-2.51(m，2H)，1.13-1.23ppm(m，6H)。MS m/z: 537.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ=8.04 (d, J=3.0Hz, 1H), 7.41-7.48 (m, 1H), 7.31-7.38 ( m, 4H), 7.07-7.12 (m, 3H), 6.92 (d, J=3.1Hz, 1H), 6.85 (d, J=8.7Hz, 2H), 4.89-5.07 (m, 2H), 3.95-4.08 (m, 2H), 2.69-2.82 (m, 1H), 2.37-2.51 (m, 2H), 1.13-1.23ppm (m, 6H).

步骤2：化合物TM05的合成Step 2: Synthesis of compound TM05

将化合物TM05_2(170mg，316.58μmol)加到甲醇(1.8mL)，四氢呋喃(1.8mL)和水(0.6mL)，然后加入一水合氢氧化锂(53.14mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝30％-60％，8.0min)纯化得到化合物TM05。MS m/z：509.3[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 8.13-8.19(m，1H)7.97(br s，1H)7.84(td，J＝8.44，3.13Hz，1H)7.45-7.51(m，2H)7.38-7.43(m，2H)7.33(br s，2H)7.12(br d，J＝8.63Hz，3H)5.36-5.49(m，2H)2.57-2.70(m，2H)2.25-2.35(m，1H)1.06(br d，J＝6.75Hz，3H).Compound TM05_2 (170 mg, 316.58 μmol) was added to methanol (1.8 mL), tetrahydrofuran (1.8 mL) and water (0.6 mL), and then lithium hydroxide monohydrate (53.14 mg) was added, and stirred at 25°C for 12 hours. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2M), and ethyl acetate (5 mL×3) was added for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM05 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%=30%-60%, 8.0min). MS m/z: 509.3 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 8.13-8.19 (m, 1H) 7.97 (br s, 1H) 7.84 (td, J=8.44, 3.13Hz, 1H) 7.45-7.51 (m, 2H) 7.38-7.43 (m, 2H) 7.33 (br s, 2H) 7.12 (br d, J＝8.63Hz, 3H) 5.36-5.49 (m, 2H) 2.57-2.70 (m, 2H) 2.25-2.35 (m, 1H) 1.06 (br d, J＝6.75Hz, 3H).

实施例5
Example 5

步骤1：化合物TM06_2的合成Step 1: Synthesis of compound TM06_2

将化合物TM01_5(540mg，1.37mmol)加到叔丁醇(6mL)中，然后加入化合物TM06_1(452.58mg，2.05mmol)和乙酸(1.16mL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(10mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和食盐水(20mL×3)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM06_2。MSm/z：553.1[M+1]+.1H NMR(400MHz，CDCl3)：δ＝7.52(s，2H)，7.31-7.39(m，4H)，7.10-7.15(m，2H)，7.02-7.05(m，1H)，6.83-6.89(m，2H)，6.77(d，J＝8.2Hz，1H)，4.90-5.07(m，2H)，3.97-4.09(m，2H)，2.73-2.82(m，1H)，2.39-2.51(m，2H)，1.15-1.23ppm(m，6H).Compound TM01_5 (540 mg, 1.37 mmol) was added to tert-butyl alcohol (6 mL), followed by compound TM06_1 (452.58 mg, 2.05 mmol) and acetic acid (1.16 mL), stirred at 115°C for 12 hr, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (10 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM06_2. MSm/z: 553.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ=7.52 (s, 2H), 7.31-7.39 (m, 4H), 7.10-7.15 (m, 2H), 7.02-7.05 (m, 1H), 6.83-6.89 (m, 2H), 6.77 (d, J=8. 2Hz, 1H), 4.90-5.07(m, 2H), 3.97-4.09(m, 2H), 2.73-2.82(m, 1H), 2.39-2.51(m, 2H), 1.15-1.23ppm(m, 6H).

步骤2：化合物TM06的合成Step 2: Synthesis of compound TM06

将化合物TM06_2(130mg，234.90μmol)加到甲醇(1.5mL)，四氢呋喃(1.5mL)和水(0.5mL)，然后加入一水合氢氧化锂(39.43mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：(ACN％＝30％-65％，8.0min)纯化得到化合物TM06。MS m/z：525.3[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 7.85-7.94(m，2H)7.48(d，J＝8.50Hz，2H)7.28-7.43(m，4H)7.24(d，J＝7.63Hz，1H)7.16(br d，J＝8.38Hz，2H)7.01(d，J＝8.13Hz，1H)5.27-5.42(m，2H)2.63-2.71(m，1H)2.58(brdd，J＝13.45，6.94Hz，1H)2.24-2.33(m，1H)1.05(d，J＝6.88Hz，3H)Compound TM06_2 (130 mg, 234.90 μmol) was added to methanol (1.5 mL), tetrahydrofuran (1.5 mL) and water (0.5 mL), and then lithium hydroxide monohydrate (39.43 mg) was added, and stirred at 25°C for 12 hours. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2M), and ethyl acetate (5 mL×3) was added for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM06 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobility: [water (0.04% HCl)-ACN]; gradient: (ACN%=30%-65%, 8.0min). MS m/z: 525.3 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 7.85-7.94 (m, 2H) 7.48 (d, J=8.50 Hz, 2H) 7.28-7.43 (m, 4H) 7.24 (d, J=7.63 Hz, 1H) 7.16 (br d, J＝8.38Hz, 2H) 7.01 (d, J＝8.13Hz, 1H) 5.27-5.42 (m, 2H) 2.63-2.71 (m, 1H) 2.58 (brdd, J＝13.45, 6.94Hz, 1H) 2.24-2.33 (m, 1H) 1.05 (d, J＝6.88Hz, 3H)

实施例6
Example 6

步骤1：化合物TM07_2的合成Step 1: Synthesis of compound TM07_2

将化合物TM01_5(540mg，1.37mmol)加到叔丁醇(6mL)中，然后加入化合物TM07_1(452.58mg，2.05mmol)和乙酸(1.16mL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(10mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和食盐水(20mL×3)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM07_2。MSm/z：553.1[M+1]+.1H NMR(400MHz，CDCl3)：δ＝8.13(d，J＝2.6Hz，1H)，7.64(q，J＝3.0Hz，1H)，7.52-7.56(m，1H)，7.31-7.38(m，4H)，7.09-7.11(m，2H)，6.88(dd，J＝8.7，5.7Hz，3H)，4.91-5.10(m，2H)，3.95-4.08(m，2H)，2.71-2.83(m，1H)，2.38-2.53(m，2H)，1.15-1.23ppm(m，6H).Compound TM01_5 (540 mg, 1.37 mmol) was added to tert-butyl alcohol (6 mL), followed by compound TM07_1 (452.58 mg, 2.05 mmol) and acetic acid (1.16 mL), stirred at 115°C for 12 hr, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (10 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM07_2. MSm/z: 553.1[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ=8.13 (d, J=2.6Hz, 1H), 7.64 (q, J=3.0Hz, 1H), 7.52-7.56 (m, 1H), 7.31-7.38 (m, 4H), 7.09-7.11 (m, 2H), 6. 88(dd, J=8.7, 5.7Hz, 3H), 4.91-5.10(m, 2H), 3.95-4.08(m, 2H), 2.71-2.83(m, 1H), 2.38-2.53(m, 2H), 1.15-1.23ppm(m, 6H).

步骤2：化合物TM07的合成Step 2: Synthesis of compound TM07

将化合物TM07_2(200mg，361.38μmol)加到甲醇(1.8mL)，四氢呋喃(1.8mL)和水(0.6mL)，然后加入一水合氢氧化锂(60.65mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：(ACN％＝25％-60％，8.0min)纯化得到化合物TM07。MS m/z：525.3[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 8.15-8.26(m，1H)7.96(br d，J＝8.63Hz，1H)7.82-7.90(m，1H)7.47(br d，J＝7.75Hz，2H)7.38(br d，J＝7.63Hz，2H)7.21-7.34(m，2H)7.08-7.15(m，3H)5.36(br s，2H)2.63-2.71(m，1H)2.54-2.61(m，1H)2.21-2.33(m，1H)1.03-1.08(m，3H).Compound TM07_2 (200 mg, 361.38 μmol) was added to methanol (1.8 mL), tetrahydrofuran (1.8 mL) and water (0.6 mL), and then lithium hydroxide monohydrate (60.65 mg) was added, and stirred at 25°C for 12 hours. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2M), and ethyl acetate (5 mL×3) was added for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM07 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobility: [water (0.04% HCl)-ACN]; gradient: (ACN%=25%-60%, 8.0min). MS m/z: 525.3 [M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δ ppm 8.15-8.26 (m, 1H) 7.96 (br d, J=8.63 Hz, 1H) 7.82-7.90 (m, 1H) 7.47 (br d, J=7.75 Hz, 2H) 7.38 (br d, J=7.63 Hz, 2H) 7.21-7.34 (m, 2H) 7.08-7.15 (m, 3H) 5.36 (br d, J=7. s, 2H) 2.63-2.71 (m, 1H) 2.54-2.61 (m, 1H) 2.21-2.33 (m, 1H) 1.03-1.08 (m, 3H).

实施例7
Example 7

步骤1：化合物TM08_2的合成Step 1: Synthesis of compound TM08_2

将化合物TM08_1(1g，6.24mmol)加到乙醇(10mL)中，然后滴入浓硫酸(499.20μL，浓度：98％)，80℃搅拌12hr。反应结束，减压浓缩除去溶剂。浓缩物加水(10mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和碳酸氢钠(20mL×3)溶液洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂，得到化合物TM08_2，直接用于下一步。MS m/z：217.3[M+1]+.1H NMR(400MHz，CDCl3)：δ＝4.12(q，J＝7.1Hz，4H)，2.23-2.33(m，2H)，1.82-1.92(m，2H)，1.23-1.28(m，6H)，1.18ppm(s，6H).Compound TM08_1 (1 g, 6.24 mmol) was added to ethanol (10 mL), and then concentrated sulfuric acid (499.20 μL, concentration: 98%) was added dropwise, and stirred at 80°C for 12 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The concentrate was diluted with water (10 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, washed with saturated sodium bicarbonate solution (20 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent to obtain compound TM08_2, which was directly used in the next step. MS m/z: 217.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ=4.12 (q, J=7.1Hz, 4H), 2.23-2.33 (m, 2H), 1.82-1.92 (m, 2H), 1.23-1.28 (m, 6H), 1.18ppm (s, 6H).

步骤2：化合物TM08_3的合成Step 2: Synthesis of compound TM08_3

向四氢呋喃(10mL)中加入叔丁醇钾(1M四氢呋喃溶液，10.91mL)，控制温度不超过30℃，将体系降至0℃，然后向体系中加入溶于四氢呋喃(2mL)的化合物TM08_2(1.18g，5.46mmol)和甲酸乙酯(658.27μL，8.18mmol)溶液，滴加过程控制温度不超过10℃，然后将温度升至20℃-25℃搅拌3hr。反应结束，减压浓缩得粗产品，然后将粗品溶于水(6mL)中得到TM08_3的水溶液，直接用于下一步。MS m/z：245.3[M+1]+.Add potassium tert-butoxide (1M tetrahydrofuran solution, 10.91 mL) to tetrahydrofuran (10 mL), control the temperature not to exceed 30°C, cool the system to 0°C, then add compound TM08_2 (1.18 g, 5.46 mmol) and ethyl formate (658.27 μL, 8.18 mmol) solution dissolved in tetrahydrofuran (2 mL) to the system, control the temperature not to exceed 10°C during the dropwise addition, then raise the temperature to 20°C-25°C and stir for 3 hr. After the reaction is completed, concentrate under reduced pressure to obtain a crude product, then dissolve the crude product in water (6 mL) to obtain an aqueous solution of TM08_3, which is directly used in the next step. MS m/z: 245.3[M+1] + .

步骤3：化合物TM08_4的合成Step 3: Synthesis of compound TM08_4

将化合物S-乙基异硫脲氢溴酸盐(927.08mg，5.01mmol)加到上一步TM08_3的水溶液中，40℃搅拌12hr。将反应体系降至室温，用乙酸乙酯(20mL×3)萃取，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝3∶1)，得到化合物TM08_4。MS m/z：285.3[M+1]+.1H NMR(400MHz，CDCl3)：δ＝10.03-10.30(m，1H)，7.66(s，1H)，4.11-4.17(m，2H)，3.14-3.22(m，2H)，2.70-2.73(m，2H)，1.36-1.43(m，3H)，1.27(t，J＝7.1Hz，3H)，1.23ppm(s，6H).Compound S-ethylisothiourea hydrobromide (927.08 mg, 5.01 mmol) was added to the aqueous solution of TM08_3 in the previous step and stirred at 40°C for 12 hours. The reaction system was cooled to room temperature and extracted with ethyl acetate (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=3:1) to obtain compound TM08_4. MS m/z: 285.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ=10.03-10.30 (m, 1H), 7.66 (s, 1H), 4.11-4.17 (m, 2H), 3.14-3.22 (m, 2H), 2.70-2.73 (m, 2H), 1.36-1. 43(m, 3H), 1.27(t, J=7.1Hz, 3H), 1.23ppm(s, 6H).

步骤4：化合物TM08_5的合成Step 4: Synthesis of compound TM08_5

将化合物TM08_4(104mg，365.72μmol)加到二氯甲烷(1mL)中，加入二异丙基乙胺(161.07μL)，然后加入对氯苄溴(75.15mg，365.72μmol)，25℃搅拌12hr。减压浓缩，粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM08_5。MS m/z：409.3[M+1]+.1H NMR(400MHz，CDCl3)：δ＝7.39(d，J＝8.5Hz，2H)，7.19-7.21(m，1H)，7.15(d，J＝8.3Hz，2H)，4.96(s，2H)，3.93-4.01(m，2H)，3.26-3.34(m，2H)，2.70-2.73(m，2H)，1.34-1.41(m，3H)，1.16-1.23ppm(m，9H)Compound TM08_4 (104 mg, 365.72 μmol) was added to dichloromethane (1 mL), followed by diisopropylethylamine (161.07 μL) and then p-chlorobenzyl bromide (75.15 mg, 365.72 μmol), and stirred at 25° C. for 12 hr. The mixture was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM08_5. MS m/z: 409.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ=7.39 (d, J=8.5Hz, 2H), 7.19-7.21 (m, 1H), 7.15 (d, J=8.3Hz, 2H), 4.96 (s, 2H), 3.93-4.01 (m, 2H), 3.26- 3.34(m, 2H), 2.70-2.73(m, 2H), 1.34-1.41(m, 3H), 1.16-1.23ppm(m, 9H)

步骤5：化合物TM08_6的合成Step 5: Synthesis of compound TM08_6

将TM08_5(540mg，1.32mmol)加到叔丁醇(5mL)中，然后加入TM01_5(368.83mg，1.98mmol)和乙酸(1.12mL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM08_6。MS m/z：533.3[M+1]+.1H NMR(400MHz，CDCl3)δ(ppm)＝7.41-7.39(m，1H)，7.39-7.35(m，2H)，7.19-7.14(m，2H)，4.99-4.96(m，2H)，4.02-3.95(m，2H)，3.31-3.23(m，2H)，2.71-2.69(m，2H)，1.36(t，J＝7.4Hz，3H)，1.25-1.21(m，2H)，1.13(t，J＝7.1Hz，3H)，1.09-1.04(m，2H).TM08_5 (540 mg, 1.32 mmol) was added to tert-butyl alcohol (5 mL), followed by TM01_5 (368.83 mg, 1.98 mmol) and acetic acid (1.12 mL), stirred at 115°C for 12 hr, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM08_6. MS m/z: 533.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ (ppm) = 7.41-7.39 (m, 1H), 7.39-7.35 (m, 2H), 7.19-7.14 (m, 2H), 4.99-4.96 (m, 2H), 4.02-3.95 (m, 2H), 3.3 1-3.23 (m, 2H), 2.71-2.69 (m, 2H), 1.36 (t, J=7.4Hz, 3H), 1.25-1.21 (m, 2H), 1.13 (t, J=7.1Hz, 3H), 1.09-1.04 (m, 2H).

步骤6：化合物TM08的合成Step 6: Synthesis of compound TM08

向TM08_06(80mg，150.09μmol)中加入甲醇(0.6mL)，四氢呋喃(0.6mL)和水(0.2mL)，然后加 入一水合氢氧化锂(25.19mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝20％-55％，8.0min)纯化得到TM08。MS m/z：505.3[M+1]+，1H NMR(400MHz，DMSO-d6)δppm 8.12-8.20(m，1H)7.98-8.04(m，1H)7.83(s，1H)7.48-7.53(m，2H)7.42-7.47(m，2H)7.33-7.41(m，2H)7.12-7.21(m，3H)7.02-7.09(m，1H)5.44-5.56(m，2H)2.59(br s，2H)1.08(s，6H)。Methanol (0.6 mL), tetrahydrofuran (0.6 mL) and water (0.2 mL) were added to TM08_06 (80 mg, 150.09 μmol), and then Lithium hydroxide monohydrate (25.19 mg) was added and stirred at 25°C for 12 hours. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2M), and ethyl acetate (5 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. TM08 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%=20%-55%, 8.0min). MS m/z: 505.3 [M+1] + , 1 H NMR (400MHz, DMSO-d6) δppm 8.12-8.20 (m, 1H) 7.98-8.04 (m, 1H) 7.83 (s, 1H) 7.48-7.53 (m, 2H) 7.42-7.47 (m, 2H) 7.33-7.41 (m, 2H) 7.12-7.21 (m, 3H) 7.02-7.09 (m, 1H) 5.44-5.56 (m, 2H) 2.59 (br s, 2H) 1.08 (s, 6H).

实施例8
Example 8

步骤1：化合物TM09_1的合成Step 1: Synthesis of compound TM09_1

将化合物TM08_5(100mg，244.53μmol)加到叔丁醇(2mL)中，然后加入化合物TM04_1(74.90mg，366.80μmol)和乙酸(207.74μL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM09_1。MS m/z：551.3[M+1]+.1H NMR(400MHz，CDCl3)：δ＝7.66-7.79(m，2H)，7.35-7.40(m，2H)，7.10-7.13(m，2H)，6.93-6.98(m，2H)，6.84-6.91(m，2H)，6.70-6.74(m，2H)，4.99(s，2H)，3.96-4.04(m，2H)，2.52-2.56(m，2H)，1.22(t，J＝7.2Hz，3H)，1.18ppm(s，6H).步骤2：化合物TM09的合成Compound TM08_5 (100 mg, 244.53 μmol) was added to tert-butyl alcohol (2 mL), followed by compound TM04_1 (74.90 mg, 366.80 μmol) and acetic acid (207.74 μL), stirred at 115°C for 12 hours, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM09_1. MS m/z: 551.3 [M+1] + . 1 H NMR (400 MHz, CDCl 3 ): δ = 7.66-7.79 (m, 2H), 7.35-7.40 (m, 2H), 7.10-7.13 (m, 2H), 6.93-6.98 (m, 2H), 6.84-6.91 (m, 2H), 6.70-6.74 (m, 2H), 4.99 (s, 2H), 3.96-4.04 (m, 2H), 2.52-2.56 (m, 2H), 1.22 (t, J = 7.2 Hz, 3H), 1.18 ppm (s, 6H). Step 2: Synthesis of compound TM09

向化合物TM09_1(130mg，235.93μmol)中加入甲醇(1.5mL)，四氢呋喃(1.5mL)和水(0.5mL)，然后加入一水合氢氧化锂(39.60mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝15％-50％，8.0min)纯化得到TM09。MS m/z：523.3[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 9.01-9.09(m，1H)8.02(q，J＝8.17Hz，1H)7.77-7.86(m，1H)7.45-7.51(m，2H)7.39-7.43(m，2H)7.25-7.38(m，2H)7.16(br d，J＝8.50Hz，2H)6.90-6.95(m，1H)6.88(dd，J＝7.88，2.13Hz，1H)5.33-5.47(m，2H)2.53-2.57(m，2H)1.06(s，6H).Methanol (1.5 mL), tetrahydrofuran (1.5 mL) and water (0.5 mL) were added to compound TM09_1 (130 mg, 235.93 μmol), and then lithium hydroxide monohydrate (39.60 mg) was added, and stirred at 25°C for 12 hours. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2M), and ethyl acetate (5 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. TM09 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%＝15%-50%, 8.0min). MS m/z：523.3[M+1] + . 1 H NMR (400MHz, DMSO-d 6 )δppm 9.01-9.09 (m，1H) 8.02 (q，J＝8.17Hz，1H) 7.77-7.86 (m，1H) 7.45-7.51 (m，2H) 7.39-7.43 (m，2H) 7.25-7.38 (m，2H) 7.16 (br d, J=8.50Hz, 2H) 6.90-6.95 (m, 1H) 6.88 (dd, J=7.88, 2.13Hz, 1H) 5.33-5.47 (m, 2H) 2.53-2.57 (m, 2H) 1.06 (s, 6H).

实施例9
Example 9

步骤1：化合物TM10_1的合成Step 1: Synthesis of compound TM10_1

将化合物TM8_5(100mg，244.53μmol)加到叔丁醇(2mL)中，然后加入化合物TM05_1(74.90mg，366.80μmol)和乙酸(207.74μL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到TM10_1。MS m/z：551.4[M+1]+.1H NMR(400MHz，CDCl3)：δ＝8.02(d，J＝2.5Hz，2H)，7.36-7.38(m，2H)，7.10(s，2H)，6.92(s，2H)，6.85(br s，2H)，6.73(s，2H)，4.98(s，2H)，3.96-4.03(m，2H)，2.53(s，2H)，1.22(t，J＝7.1Hz，3H)，1.18ppm(s，6H).Compound TM8_5 (100 mg, 244.53 μmol) was added to tert-butyl alcohol (2 mL), followed by compound TM05_1 (74.90 mg, 366.80 μmol) and acetic acid (207.74 μL), stirred at 115°C for 12 hours, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain TM10_1. MS m/z: 551.4[M+1] + . 1 H NMR (400MHz, CDCl 3 ): δ=8.02 (d, J=2.5Hz, 2H), 7.36-7.38 (m, 2H), 7.10 (s, 2H), 6.92 (s, 2H), 6.85 (br s, 2H), 6.73 (s, 2H), 4.98 (s , 2H), 3.96-4.03 (m, 2H), 2.53 (s, 2H), 1.22 (t, J=7.1Hz, 3H), 1.18ppm (s, 6H).

步骤2：化合物TM10的合成Step 2: Synthesis of compound TM10

向化合物TM10_1(170mg，308.53μmol)中加入甲醇(1.5mL)，四氢呋喃(1.5mL)和水(0.5mL)， 然后加入一水合氢氧化锂(51.78mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝15％-50％，8.0min)纯化得到TM10。MS m/z：523.3[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 8.13-8.18(m，1H)7.88-7.95(m，1H)7.84(td，J＝8.47，3.06Hz，1H)7.47-7.52(m，2H)7.39-7.45(m，2H)7.26-7.39(m，2H)7.10-7.17(m，3H)5.37-5.52(m，2H)2.57(s，2H)1.07(s，6H).Methanol (1.5 mL), tetrahydrofuran (1.5 mL) and water (0.5 mL) were added to compound TM10_1 (170 mg, 308.53 μmol). Then lithium hydroxide monohydrate (51.78 mg) was added and stirred at 25°C for 12 hours. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2M), and ethyl acetate (5 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. TM10 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%=15%-50%, 8.0min). MS m/z: 523.3[M+1] + . 1 H NMR (400MHz, DMSO-d 6 )δppm 8.13-8.18 (m, 1H) 7.88-7.95 (m, 1H) 7.84 (td, J=8.47, 3.06Hz, 1H) 7.47-7.52 (m, 2H) 7.39-7.45 (m, 2H) 7.26-7.39 (m, 2H) 7.10-7.17 (m, 3H) 5.37-5. 52(m,2H)2.57(s,2H)1.07(s,6H).

实施例10
Example 10

步骤1：化合物TM11_1的合成Step 1: Synthesis of compound TM11_1

将TM08_5(100mg，244.53μmol)加到叔丁醇(2mL)中，然后加入化合物TM06_1(80.94mg，366.80μmol)和乙酸(207.74μL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM11_1。MS m/z：567.3[M+1]+.1HNMR(400MHz，CDCl3)：δ＝7.54-7.65(m，2H)，7.36-7.40(m，2H)，7.10-7.13(m，2H)，6.95(d，J＝8.7Hz，2H)，6.84-6.90(m，2H)，6.71(d，J＝8.7Hz，2H)，4.99(s，2H)，4.00(q，J＝7.1Hz，2H)，2.54(s，2H)，1.22(t，J＝7.1Hz，3H)，1.18ppm(s，6H).TM08_5 (100 mg, 244.53 μmol) was added to tert-butyl alcohol (2 mL), followed by compound TM06_1 (80.94 mg, 366.80 μmol) and acetic acid (207.74 μL), stirred at 115°C for 12 hours, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM11_1. MS m/z: 567.3[M+1] + . 1 HNMR (400MHz, CDCl 3 ): δ=7.54-7.65 (m, 2H), 7.36-7.40 (m, 2H), 7.10-7.13 (m, 2H), 6.95 (d, J=8.7Hz, 2H), 6.84-6.90 (m, 2H), 6.7 1 (d, J=8.7Hz, 2H), 4.99 (s, 2H), 4.00 (q, J=7.1Hz, 2H), 2.54 (s, 2H), 1.22 (t, J=7.1Hz, 3H), 1.18ppm (s, 6H).

步骤2：化合物TM11的合成Step 2: Synthesis of compound TM11

向TM11_1(140mg，246.71μmol)中加入甲醇(1.5mL)，四氢呋喃(1.5mL)和水(0.5mL)，然后加入一水合氢氧化锂(41.41mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝20％-55％，8.0min)纯化得到化合物TM11。MS m/z：539.3[M+1]+，1H NMR(400MHz，DMSO-d6)δppm 7.90(t，J＝7.94Hz，1H)7.83(br s，1H)7.47-7.50(m，2H)7.39-7.42(m，2H)7.27-7.38(m，2H)7.25(d，J＝7.63Hz，1H)7.17(br d，J＝8.25Hz，2H)7.02(d，J＝8.00Hz，1H)5.31-5.42(m，2H)2.55(br s，2H)1.07(s，6H).Methanol (1.5 mL), tetrahydrofuran (1.5 mL) and water (0.5 mL) were added to TM11_1 (140 mg, 246.71 μmol), and then lithium hydroxide monohydrate (41.41 mg) was added and stirred at 25°C for 12 hr. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2 M), and ethyl acetate (5 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM11 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%=20%-55%, 8.0min). MS m/z: 539.3 [M+1] + , 1 H NMR (400MHz, DMSO-d 6 ) δppm 7.90 (t, J=7.94Hz, 1H) 7.83 (br s, 1H) 7.47-7.50 (m, 2H) 7.39-7.42 (m, 2H) 7.27-7.38 (m, 2H) 7.25 (d, J=7.63Hz, 1H) 7.17 (br d, J=8.25Hz, 2H) 7.02 (d, J=8.00Hz, 1H) 5.31-5.42 (m, 2H) 2.55 (br s, 2H) 1.07 (s, 6H).

实施例11
Embodiment 11

步骤1：化合物TM12_1的合成Step 1: Synthesis of compound TM12_1

将化合物TM8_5(100mg，244.53μmol)加到叔丁醇(2mL)中，然后加入化合物TM07_1(80.94mg，366.80μmol)和乙酸(207.74μL)，115℃搅拌12hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到TM12_1。MS m/z：567.3[M+1]+，1H NMR(400MHz，CDCl3)：δ＝7.62(ddd，J＝17.0，8.8，2.6Hz，2H)，7.35-7.39(m，2H)，7.10-7.12(m，2H)，6.93(d，J＝8.6Hz，2H)，6.84-6.88(m，2H)，6.69-6.74(m，2H)，4.96-5.00(m，2H)，4.00(q，J＝7.0Hz，2H)，2.54(s，2H)，1.22(t，J＝7.1Hz，3H)，1.18ppm(s，6H)Compound TM8_5 (100 mg, 244.53 μmol) was added to tert-butyl alcohol (2 mL), followed by compound TM07_1 (80.94 mg, 366.80 μmol) and acetic acid (207.74 μL), stirred at 115°C for 12 hours, the reaction system was cooled to room temperature, diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (10 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain TM12_1. MS m/z: 567.3[M+1] + , 1 H NMR (400MHz, CDCl 3 ): δ=7.62 (ddd, J=17.0, 8.8, 2.6Hz, 2H), 7.35-7.39 (m, 2H), 7.10-7.12 (m, 2H), 6.93 (d, J=8.6Hz, 2H), 6.84-6.8 8 (m, 2H), 6.69-6.74 (m, 2H), 4.96-5.00 (m, 2H), 4.00 (q, J=7.0Hz, 2H), 2.54 (s, 2H), 1.22 (t, J=7.1Hz, 3H), 1.18ppm (s, 6H)

步骤2：化合物TM12的合成Step 2: Synthesis of compound TM12

将化合物TM12_1(60.00mg，105.73μmol)中加入甲醇(0.6mL)，四氢呋喃(0.6mL)和水(0.2mL)， 然后加入一水合氢氧化锂(17.75mg)，25℃搅拌12hr。反应结束，用稀盐酸(2M)调pH＝3～4，加乙酸乙酯(5mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝25％-60％，8.0min)纯化得到化合物TM12。MS m/z：539.3[M+1]+，1H NMR(400MHz，DMSO-d6)δppm 8.15-8.19(m，1H)8.04(s，1H)7.90-7.96(m，1H)7.44(s，4H)7.39(br d，J＝8.25Hz，2H)7.13(br d，J＝8.51Hz，2H)7.05-7.10(m，1H)5.58-5.66(m，2H)2.53-2.60(m，2H)1.04(s，6H).Methanol (0.6 mL), tetrahydrofuran (0.6 mL) and water (0.2 mL) were added to compound TM12_1 (60.00 mg, 105.73 μmol). Then, lithium hydroxide monohydrate (17.75 mg) was added and stirred at 25°C for 12 hours. After the reaction was completed, the pH was adjusted to 3-4 with dilute hydrochloric acid (2M), and ethyl acetate (5 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM12 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%=25%-60%, 8.0min). MS m/z: 539.3 [M+1] + , 1 H NMR (400MHz, DMSO-d 6 ) δ ppm 8.15-8.19 (m, 1H) 8.04 (s, 1H) 7.90-7.96 (m, 1H) 7.44 (s, 4H) 7.39 (br d, J=8.25Hz, 2H) 7.13 (br d, J=8.51Hz, 2H) 7.05-7.10 (m, 1H) 5.58-5.66 (m, 2H) 2.53-2.60 (m, 2H) 1.04 (s, 6H).

实施例12
Example 12

步骤1：化合物TM13_2的合成Step 1: Synthesis of compound TM13_2

将TM13_1(40.42g，199.77mmol，49.29mL)缓慢滴加到氮气保护下的三正丁基膦(100g，998.85mmol，108.58mL)中，反应在60℃下搅拌2hr。反应完毕，减压旋除残余原料后湿法上样，快速硅胶柱(石油醚∶乙酸乙酯＝100∶1-10∶1)纯化，得到化合物TM13_2，1H NMR(400MHz，CDCl3，299K)δ(ppm)＝6.22-6.18(m，1H)，5.62-5.57(m，1H)，4.25-4.19(m，2H)，4.18-4.10(m，2H)，2.68-2.61(m，2H)，2.55-2.49(m，2H)，1.34-1.29(m，3H)，1.28-1.22(m，3H).TM13_1 (40.42 g, 199.77 mmol, 49.29 mL) was slowly added dropwise to tri-n-butylphosphine (100 g, 998.85 mmol, 108.58 mL) under nitrogen protection, and the reaction was stirred at 60°C for 2 hr. After the reaction was completed, the residual raw material was removed under reduced pressure and then wet-loaded and purified by flash silica gel column (petroleum ether: ethyl acetate = 100: 1-10: 1) to obtain compound TM13_2, 1 H NMR (400 MHz, CDCl 3 , 299K) δ (ppm) = 6.22-6.18 (m, 1H), 5.62-5.57 (m, 1H), 4.25-4.19 (m, 2H), 4.18-4.10 (m, 2H), 2.68-2.61 (m, 2H), 2.55-2.49 (m, 2H), 1.34-1.29 (m, 3H), 1.28-1.22 (m, 3H).

步骤2：化合物TM13_3的合成Step 2: Synthesis of compound TM13_3

向干燥三颈瓶中加入三甲基碘化亚砜(29.54g，134.21mmol)，加入二甲基亚砜(120mL)溶解，然后缓慢加入钠氢(5.37g，134.21mmol，60％纯度)，在20℃搅拌1hr，然后0℃冰浴控温(外温)下缓慢滴加溶解在二甲基亚砜(120mL)中的TM13_2(24.43g，122.01mmol)，滴加完毕后在15℃反应16h。反应结束，缓慢加入150mL饱和氯化铵溶液淬灭反应，乙酸乙酯(160mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤并减压旋干。硅胶吸附柱(石油醚∶乙酸乙酯＝100∶0-10∶1)纯化，得到化合物TM13_3，1H NMR(400MHz，CDCl3，297K)δ(ppm)＝4.12(dq，J＝3.4，7.1Hz，4H)，2.56-2.46(m，2H)，1.92-1.80(m，2H)，1.28-1.20(m，8H)，0.77-0.69(m，2H).Add trimethyl sulfoxide iodide (29.54 g, 134.21 mmol) to a dry three-necked bottle, add dimethyl sulfoxide (120 mL) to dissolve, then slowly add sodium hydrogen (5.37 g, 134.21 mmol, 60% purity), stir at 20 ° C for 1 hour, then slowly drop TM13_2 (24.43 g, 122.01 mmol) dissolved in dimethyl sulfoxide (120 mL) under 0 ° C ice bath temperature control (external temperature), react at 15 ° C for 16 hours after the addition is complete. After the reaction is completed, slowly add 150 mL of saturated ammonium chloride solution to quench the reaction, extract with ethyl acetate (160 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter and spin dry under reduced pressure. Purification by silica gel adsorption column (petroleum ether: ethyl acetate = 100: 0-10: 1) gave compound TM13_3, 1 H NMR (400 MHz, CDCl 3 , 297K) δ (ppm) = 4.12 (dq, J = 3.4, 7.1 Hz, 4H), 2.56-2.46 (m, 2H), 1.92-1.80 (m, 2H), 1.28-1.20 (m, 8H), 0.77-0.69 (m, 2H).

步骤3：化合物TM13_4的合成Step 3: Synthesis of compound TM13_4

将叔丁醇钾的四氢呋喃溶液(1M，49.47mL)加入到干燥后的三颈瓶中，降温至0℃，再慢慢依次滴加溶解在四氢呋喃(40mL)中的TM13_3(5.3g，24.74mmol)和溶解在四氢呋喃(15mL)中的甲酸乙酯(2.75g，37.10mmol，2.98mL)，滴加完毕后，反应在20℃下搅拌2小时。加入水(40mL)，分液，水相再次用乙酸乙酯(40mL)萃取，合并有机相，加入5％氢氧化钠(20mL)水溶液，分液，合并两次的有机相，得到化合物TM13_4水相直接用于下一步反应。MS m/z：243.1[M+1]+.Add a tetrahydrofuran solution of potassium tert-butoxide (1M, 49.47 mL) to a dried three-necked flask, cool to 0°C, and then slowly drop TM13_3 (5.3 g, 24.74 mmol) dissolved in tetrahydrofuran (40 mL) and ethyl formate (2.75 g, 37.10 mmol, 2.98 mL) dissolved in tetrahydrofuran (15 mL) in sequence. After the addition is complete, stir the reaction at 20°C for 2 hours. Add water (40 mL), separate the liquids, extract the aqueous phase with ethyl acetate (40 mL) again, combine the organic phases, add a 5% sodium hydroxide (20 mL) aqueous solution, separate the liquids, combine the two organic phases, and obtain the compound TM13_4. The aqueous phase is directly used for the next step. MS m/z: 243.1[M+1] + .

步骤4：化合物TM13_5的合成Step 4: Synthesis of compound TM13_5

将S-乙基异硫脲氢溴酸盐(4.21g，22.75mmol)加入到化合物TM13_4(5.99g，24.72mmol)的水溶液中，反应在60℃下搅拌16小时。反应结束，加入水(40mL)，分液，水相用乙酸乙酯(40mL)萃取两次，合并有机相，无水硫酸钠干燥后拌样。快速硅胶柱(石油醚∶乙酸乙酯＝100∶0-6∶4)纯化。得到化合物TM13_5。MS m/z：283.1[M+1]+，1H NMR(400MHz，CDCl3，296K)δ(ppm)＝11.93-11.74(m，1H)，7.94-7.92(m，1H)，4.15-4.07(m，2H)，3.23-3.14(m，2H)，2.79-2.73(m，2H)，1.42-1.36(m，3H)，1.31-1.27(m，2H)，1.25-1.20(m，3H)，1.04-0.99(m，2H). S-ethylisothiourea hydrobromide (4.21 g, 22.75 mmol) was added to an aqueous solution of compound TM13_4 (5.99 g, 24.72 mmol), and the reaction was stirred at 60°C for 16 hours. After the reaction was completed, water (40 mL) was added, and the liquid was separated. The aqueous phase was extracted twice with ethyl acetate (40 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, and then mixed. Purification was performed on a rapid silica gel column (petroleum ether: ethyl acetate = 100: 0-6: 4). Compound TM13_5 was obtained. MS m/z: 283.1[M+1] + , 1 H NMR (400MHz, CDCl 3 , 296K) δ (ppm) = 11.93-11.74 (m, 1H), 7.94-7.92 (m, 1H), 4.15-4.07 (m, 2H), 3.23-3.14 (m, 2H), 2.79-2.73 (m, 2H), 1.42-1.36 (m, 3H), 1.31-1.27 (m, 2H), 1.25-1.20 (m, 3H), 1.04-0.99 (m, 2H).

步骤5：化合物TM13_6的合成Step 5: Synthesis of compound TM13_6

将TM13_5(150mg，531.24μmol)溶解在二氯甲烷(1.5mL)中，再加入对氯苄溴(120.07mg，584.36μmol)和二异丙基乙胺(231.33μL)，反应体系在25℃搅拌2hr。反应结束，直接减压旋干，快速硅胶柱(石油醚∶乙酸乙酯＝10∶1-1∶1)纯化。得化合物TM13_6。MSm/z：407.1[M+1]+，1H NMR(400MHz，CDCl3)δ(ppm)＝7.41-7.39(m，1H)，7.39-7.35(m，2H)，7.19-7.14(m，2H)，4.99-4.96(m，2H)，4.02-3.95(m，2H)，3.31-3.23(m，2H)，2.71-2.69(m，2H)，1.36(t，J＝7.4Hz，3H)，1.25-1.21(m，2H)，1.13(t，J＝7.1Hz，3H)，1.09-1.04(m，2H).TM13_5 (150 mg, 531.24 μmol) was dissolved in dichloromethane (1.5 mL), and p-chlorobenzyl bromide (120.07 mg, 584.36 μmol) and diisopropylethylamine (231.33 μL) were added, and the reaction system was stirred at 25°C for 2 hours. After the reaction was completed, the mixture was directly dried under reduced pressure and purified by a rapid silica gel column (petroleum ether: ethyl acetate = 10: 1-1: 1). Compound TM13_6 was obtained. MSm/z: 407.1[M+1] + , 1 H NMR (400MHz, CDCl 3 ) δ (ppm) = 7.41-7.39 (m, 1H), 7.39-7.35 (m, 2H), 7.19-7.14 (m, 2H), 4.99-4.96 (m, 2H), 4.02-3.95 (m, 2H), 3.3 1-3.23 (m, 2H), 2.71-2.69 (m, 2H), 1.36 (t, J=7.4Hz, 3H), 1.25-1.21 (m, 2H), 1.13 (t, J=7.1Hz, 3H), 1.09-1.04 (m, 2H).

步骤6：化合物TM13_7的合成Step 6: Synthesis of compound TM13_7

在一个预先干燥的单口瓶中加入TM13_6(189mg，464.46μmol)和特戊酸(0.6mL)，随后再加入化合物TM01_5(86.49mg，464.46μmol)，反应在130℃下反应5h。反应结束，缓慢加入5mL饱和碳酸氢钠溶液淬灭反应，乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤并减压旋干，快速硅胶柱(石油醚∶乙酸乙酯＝100∶0-6∶4)纯化。得化合物TM13_7。MSm/z：531.1[M+1]+，1H NMR(400MHz，CDCl3，296K)δ(ppm)＝8.23-8.17(m，1H)，7.72-7.66(m，1H)，7.46-7.43(m，1H)，7.39-7.33(m，4H)，7.15-7.09(m，2H)，7.02-6.98(m，1H)，6.93-6.84(m，3H)，5.05-4.99(m，2H)，4.08-3.99(m，2H)，2.56-2.52(m，2H)，1.24-1.16(m，5H)，1.01-0.97(m，2H).TM13_6 (189 mg, 464.46 μmol) and pivalic acid (0.6 mL) were added to a pre-dried single-mouth bottle, followed by compound TM01_5 (86.49 mg, 464.46 μmol), and the reaction was carried out at 130°C for 5 h. After the reaction was completed, 5 mL of saturated sodium bicarbonate solution was slowly added to quench the reaction, and ethyl acetate (10 mL×3) was used for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure, and purified by rapid silica gel column (petroleum ether: ethyl acetate = 100: 0-6: 4). Compound TM13_7 was obtained. MSm/z: 531.1[M+1] + , 1 H NMR (400MHz, CDCl 3 , 296K) δ (ppm) = 8.23-8.17 (m, 1H), 7.72-7.66 (m, 1H), 7.46-7.43 (m, 1H), 7.39-7.33 (m, 4H), 7.15-7.09 (m, 2 H), 7.02-6.98(m, 1H), 6.93-6.84(m, 3H), 5.05-4.99(m, 2H), 4.08-3.99(m, 2H), 2.56-2.52(m, 2H), 1.24-1.16(m, 5H), 1.01-0.97(m, 2H).

步骤7：化合物TM13的合成Step 7: Synthesis of compound TM13

在一个预先干燥的单口瓶中加入TM13_7(0.17g，320.15μmol)和四氢呋喃(1.7mL)，甲醇(1.7mL)，水(0.35mL)，随后再加入一水合氢氧化锂(4M，656.31μL)反应在50℃下反应2h。反应结束，加入4mL水，乙酸乙酯(5mL×3)萃取，水相用2NHCl调pH至2～3，乙酸乙酯萃取(5mL×3)，无水硫酸钠干燥后减压旋干，得化合物TM13。MS m/z：503.1[M+1]+，1H NMR(400MHz，DMSO-d6)δ(ppm)＝12.17-11.57(s，1H)，8.20-8.15(s，1H)，7.85-7.78(m，1H)，7.58-7.27(m，7H)，7.13-7.08(m，1H)，7.07-7.02(m，2H)，7.01-6.94(m，1H)，5.29-5.06(m，2H)，2.58-2.53(m，2H)，1.09-1.04(m，2H)，0.85-0.81(m，2H).TM13_7 (0.17 g, 320.15 μmol) and tetrahydrofuran (1.7 mL), methanol (1.7 mL), water (0.35 mL) were added to a pre-dried single-mouth bottle, followed by the addition of lithium hydroxide monohydrate (4 M, 656.31 μL) and the reaction was carried out at 50°C for 2 h. After the reaction was completed, 4 mL of water was added, and the mixture was extracted with ethyl acetate (5 mL×3). The aqueous phase was adjusted to pH 2-3 with 2N HCl, extracted with ethyl acetate (5 mL×3), dried over anhydrous sodium sulfate, and then dried under reduced pressure to obtain compound TM13. MS m/z: 503.1[M+1] + , 1 H NMR (400MHz, DMSO-d 6 ) δ (ppm) = 12.17-11.57 (s, 1H), 8.20-8.15 (s, 1H), 7.85-7.78 (m, 1H), 7.58-7.27 (m, 7H), 7.13-7.08 (m, 1H) , 7.07-7.02(m, 2H), 7.01-6.94(m, 1H), 5.29-5.06(m, 2H), 2.58-2.53(m, 2H), 1.09-1.04(m, 2H), 0.85-0.81(m, 2H).

实施例13
Example 13

步骤1：化合物TM14_2的合成Step 1: Synthesis of compound TM14_2

将四异丙基氧钛(6.34g，22.29mmol)和四氯化钛(13.01g，68.59mmol)溶于二氯甲烷(200mL)降温至0℃后，搅拌10min后加入二异丙基乙胺(12.08g，93.46mmol)保持0℃搅拌20min后加入化合物TM14_1(20g，85.74mmol)，搅拌1hr后，滴加化合物丙烯酸叔丁酯(16.48g，128.61mmol)后恢复20℃搅拌12hr，反应结束后，将反应液倒入500mL饱和氯酯化铵水溶液中，用乙酸乙(500mL*3)萃取，饱和食盐水500mL洗涤，减压浓缩得到残余物，残余物经过柱层析分离(洗脱剂：石油醚/乙酸乙酯＝1/0-3/1，体积比)，得到化合物TM14_2。1H NMR(400MHz，CDCl3)δ＝7.27(s，5H)，4.75-4.62(m，1H)，4.23-4.09(m，2H)，3.84-3.69(m，1H)，3.39-3.24(m，1H)，2.81-2.59(m，1H)，2.37-2.26(m，2H)，2.13-1.97(m，1H)，1.85-1.72(m，1H)，1.49-1.44(m，9H)，1.20(d，J＝6.9Hz，3H).Tetraisopropyl titanium oxide (6.34 g, 22.29 mmol) and titanium tetrachloride (13.01 g, 68.59 mmol) were dissolved in dichloromethane (200 mL), cooled to 0°C, stirred for 10 min, and then diisopropylethylamine (12.08 g, 93.46 mmol) was added. The mixture was kept at 0°C and stirred for 20 min, and then compound TM14_1 (20 g, 85.74 mmol) was added. After stirring for 1 hr, compound tert-butyl acrylate (16.48 g, 128.61 mmol) was added dropwise, and the mixture was restored to 20°C and stirred for 12 hr. After the reaction was completed, the reaction solution was poured into 500 mL of saturated aqueous ammonium chloride solution, extracted with ethyl acetate (500 mL*3), washed with 500 mL of saturated brine, and concentrated under reduced pressure to obtain a residue. The residue was separated by column chromatography (eluent: petroleum ether/ethyl acetate = 1/0-3/1, volume ratio) to obtain compound TM14_2. 1 H NMR (400MHz, CDCl 3 ) δ = 7.27 (s, 5H), 4.75-4.62 (m, 1H), 4.23-4.09 (m, 2H), 3.84-3.69 (m, 1H), 3.39-3.24 (m, 1H), 2.81-2.59 (m, 1H), 2.37-2.26 (m, 2 H), 2.13-1.97 (m, 1H), 1.85-1.72 (m, 1H), 1.49-1.44 (m, 9H), 1.20 (d, J=6.9Hz, 3H).

步骤2：化合物TM14_3的合成 Step 2: Synthesis of compound TM14_3

将过氧化氢(16.94g，149.41mmol，14.36mL，30％纯度)溶于四氢呋喃(150mL)后降温至0℃后，保持0℃时，加入氢氧化锂(2.09g，49.80mmol)的水(50mL)溶液后，保持0℃滴加化合物TM14_2(15g，41.50mmol)的四氢呋喃(50mL)溶液0℃搅拌2hr。反应完毕后，向反应液中滴加饱和亚硫酸钠水溶液(150mL)淬灭反应，淀粉碘化钾试纸不变蓝，后用二氯甲烷(150mL)萃取，收集水相，水相用2M稀盐酸调节pH＝1～2，后用二氯甲烷(300mL)萃取三次，收集有机相，有机相无水硫酸钠干燥，过滤减压浓缩得到化合物TM14_3。1H NMR(400MHz，CDCl3)δ＝2.58-2.46(m，1H)，2.35-2.25(m，2H)，2.04-1.85(m，1H)，1.83-1.70(m，1H)，1.51-1.39(m，9H)，1.24-1.16(m，3H)Hydrogen peroxide (16.94 g, 149.41 mmol, 14.36 mL, 30% purity) was dissolved in tetrahydrofuran (150 mL) and then cooled to 0°C. After adding a solution of lithium hydroxide (2.09 g, 49.80 mmol) in water (50 mL) while maintaining 0°C, a solution of compound TM14_2 (15 g, 41.50 mmol) in tetrahydrofuran (50 mL) was added dropwise at 0°C and stirred for 2 hours at 0°C. After the reaction was completed, a saturated aqueous sodium sulfite solution (150 mL) was added dropwise to the reaction solution to quench the reaction, and the starch potassium iodide test paper did not turn blue. The reaction solution was then extracted with dichloromethane (150 mL), and the aqueous phase was collected. The aqueous phase was adjusted to pH 1-2 with 2M dilute hydrochloric acid, and then extracted three times with dichloromethane (300 mL). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound TM14_3. 1 H NMR (400MHz, CDCl 3 ) δ = 2.58-2.46 (m, 1H), 2.35-2.25 (m, 2H), 2.04-1.85 (m, 1H), 1.83-1.70 (m, 1H), 1.51-1.39 (m, 9H), 1.24-1.16 (m, 3H)

步骤3：化合物TM14_4的合成Step 3: Synthesis of compound TM14_4

将化合物TM14_3(8g，39.56mmol)溶于乙醇(160mL)后加入浓硫酸(5.94g，59.33mmol，98％)后80℃搅拌12hr，反应完毕后，反应液降至20～25℃，用饱和碳酸氢钠(300mL)淬灭反应，后用乙酸乙酯(500mL*3)萃取，收集有机相，有机相用饱和食盐水(300mL)洗涤，后有机相无水硫酸钠干燥，过滤减压浓缩得到化合物TM14_4。MS m/z：203.3[M+1]+.1H NMR(400MHz，CDCl3)δ＝4.20-4.08(m，4H)，2.54-2.43(m，1H)，2.38-2.26(m，2H)，2.02-1.90(m，1H)，1.84-1.72(m，1H)，1.31-1.22(m，6H)，1.20-1.14(m，3H).Compound TM14_3 (8 g, 39.56 mmol) was dissolved in ethanol (160 mL), and concentrated sulfuric acid (5.94 g, 59.33 mmol, 98%) was added, followed by stirring at 80°C for 12 hr. After the reaction was completed, the reaction solution was cooled to 20-25°C, and the reaction was quenched with saturated sodium bicarbonate (300 mL). The mixture was then extracted with ethyl acetate (500 mL*3), and the organic phase was collected and washed with saturated brine (300 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound TM14_4. MS m/z: 203.3[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ = 4.20-4.08 (m, 4H), 2.54-2.43 (m, 1H), 2.38-2.26 (m, 2H), 2.02-1.90 (m, 1H), 1.84-1.72 (m, 1H), 1.31-1. 22(m,6H),1.20-1.14(m,3H).

步骤4：化合物TM14_5的合成Step 4: Synthesis of compound TM14_5

将化合物TM14_4(9.9g，48.95mmol)和叔丁氧基双(二甲胺基)甲烷(28.44g，146.85mmol)溶于甲苯(55mL)后115℃搅拌12hr后降温至20℃后50℃减压浓缩得到残余物，加入四氢呋喃(55mL)和盐酸(1M，97.90mL)20℃搅拌3hr，反应完毕后，将反应液倒入500mL的水中，然后用饱和碳酸钠水溶液调节pH＝7～8，后用乙酸乙酯(500mL)萃取，有机相减压浓缩得到化合物TM14_5，直接用于下一步，MS m/z：231.3[M+1]+ Compound TM14_4 (9.9 g, 48.95 mmol) and tert-butoxybis(dimethylamino)methane (28.44 g, 146.85 mmol) were dissolved in toluene (55 mL), stirred at 115°C for 12 hr, cooled to 20°C, and concentrated under reduced pressure at 50°C to obtain a residue. Tetrahydrofuran (55 mL) and hydrochloric acid (1 M, 97.90 mL) were added and stirred at 20°C for 3 hr. After the reaction was completed, the reaction solution was poured into 500 mL of water, and then adjusted to pH = 7-8 with saturated sodium carbonate aqueous solution, and then extracted with ethyl acetate (500 mL). The organic phase was concentrated under reduced pressure to obtain compound TM14_5, which was directly used in the next step. MS m/z: 231.3 [M+1] +

步骤5：化合物TM14_6的合成Step 5: Synthesis of compound TM14_6

将化合物TM14_5(5g，21.71mmol)溶于水(70mL)后加入碳酸氢钠(2.74g，32.57mmol)和硫脲氢溴酸盐(4.02g，21.71mmol)后40℃搅拌12hr，将反应液用乙酸乙酯100mL和水100mL稀释，后分液，有机相减压浓缩得到残余物，残余物经过柱层析分离(洗脱剂：石油醚/乙酸乙酯＝1/0-3/1，体积比)，得到化合物TM14_6。MS m/z：271.2[M+1]+.1H NMR(400MHz，DMSO-d6)δ＝12.80-12.62(m，1H)，7.66(br s，1H)，4.08-3.93(m，2H)，3.12-3.03(m，2H)，2.82-2.67(m，1H)，2.59-2.52(m，1H)，2.43-2.29(m，1H)，1.26(t，J＝7.3Hz，3H)，1.11(t，J＝7.1Hz，3H)，1.04(d，J＝6.9Hz，3H).Compound TM14_5 (5 g, 21.71 mmol) was dissolved in water (70 mL), and sodium bicarbonate (2.74 g, 32.57 mmol) and thiourea hydrobromide (4.02 g, 21.71 mmol) were added, followed by stirring at 40°C for 12 hr. The reaction solution was diluted with 100 mL of ethyl acetate and 100 mL of water, and then the liquids were separated. The organic phase was concentrated under reduced pressure to obtain a residue. The residue was separated by column chromatography (eluent: petroleum ether/ethyl acetate = 1/0-3/1, volume ratio) to obtain compound TM14_6. MS m/z: 271.2[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δ = 12.80-12.62 (m, 1H), 7.66 (br s, 1H), 4.08-3.93 (m, 2H), 3.12-3.03 (m, 2H), 2.82-2.67 (m, 1H), 2.59-2 .52 (m, 1H), 2.43-2.29 (m, 1H), 1.26 (t, J = 7.3Hz, 3H), 1.11 (t, J = 7.1Hz, 3H), 1.04 (d, J = 6.9Hz, 3H).

步骤6：化合物TM14_7的合成Step 6: Synthesis of compound TM14_7

将化合物TM14_6(4g，14.80mmol)和化合物对氯苄溴(3.04g，14.80mmol)溶于二氯甲烷(40mL)后加入二异丙基乙胺(4.78g，36.99mmol，6.44mL)后25℃搅拌12hr将反应液用二氯甲烷50mL和水50mL稀释，后分液，有机相减压浓缩得到残余物，残余物经过柱层析分离(洗脱剂：石油醚/乙酸乙酯＝1/03/1，体积比)，得到化合物TM14_7。MS m/z：395.1[M+1]+.1H NMR(400MHz，DMSO-d6)δ＝7.79-7.73(m，1H)，7.51-7.43(m，2H)，7.29-7.21(m，2H)，5.12(br d，J＝9.5Hz，2H)，4.03-3.92(m，2H)，3.15-3.04(m，2H)，2.84-2.75(m，1H)，2.74-2.57(m，1H)，2.38-2.29(m，1H)，1.28-1.21(m，3H)，1.13-1.03(m，6H).Compound TM14_6 (4 g, 14.80 mmol) and compound p-chlorobenzyl bromide (3.04 g, 14.80 mmol) were dissolved in dichloromethane (40 mL), and diisopropylethylamine (4.78 g, 36.99 mmol, 6.44 mL) was added, and the mixture was stirred at 25°C for 12 hr. The reaction solution was diluted with 50 mL of dichloromethane and 50 mL of water, and then separated. The organic phase was concentrated under reduced pressure to obtain a residue. The residue was separated by column chromatography (eluent: petroleum ether/ethyl acetate = 1/03/1, volume ratio) to obtain compound TM14_7. MS m/z: 395.1[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δ = 7.79-7.73 (m, 1H), 7.51-7.43 (m, 2H), 7.29-7.21 (m, 2H), 5.12 (br d, J = 9.5Hz, 2H), 4.03-3.92 (m, 2H), 3 .15-3.04(m, 2H), 2.84-2.75(m, 1H), 2.74-2.57(m, 1H), 2.38-2.29(m, 1H), 1.28-1.21(m, 3H), 1.13-1.03(m, 6H).

步骤7：化合物TM14_9的合成Step 7: Synthesis of compound TM14_9

将化合物TM14_7(300.00mg，759.66μmol)和特戊酸(1.55g，15.19mmol)，和化合物TM14_8(170.65mg，911.59μmol)混合后，在130℃下反应5hr。反应结束后，加入饱和碳酸氢钠水溶液(50mL)淬灭，淬灭后体系的pH调至7～8，用乙酸乙酯(50mL*3)萃取，合并有机相，有机相用饱和食盐水(60mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩得到残余物向残余物中加入7mL(MTBE∶EA＝10∶1)搅拌1h后，过滤收集滤饼，干燥，得到化合物TM14_9，直接用于下一步，MS m/z：520.1[M+1]+.Compound TM14_7 (300.00 mg, 759.66 μmol) and pivalic acid (1.55 g, 15.19 mmol) were mixed with compound TM14_8 (170.65 mg, 911.59 μmol) and reacted at 130°C for 5 hours. After the reaction was completed, saturated sodium bicarbonate aqueous solution (50 mL) was added to quench the reaction. After quenching, the pH of the system was adjusted to 7-8, and the reaction was extracted with ethyl acetate (50 mL*3). The organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. 7 mL (MTBE: EA = 10: 1) was added to the residue and stirred for 1 hour. The filter cake was collected by filtration and dried to obtain compound TM14_9, which was used directly in the next step. MS m/z: 520.1 [M+1] + .

步骤8：化合物TM14的合成Step 8: Synthesis of compound TM14

将化合物TM14_9(350.00mg，673.10μmol)加入到四氢呋喃(6mL)，甲醇(6mL)和水(2mL)，后加入氢氧化锂(112.97mg，2.69mmol)，20℃搅拌3hr将反应液降至室温，向反应液中加入20mL水，用甲基叔丁基醚10mL萃取，有机相丢弃，用1M稀盐酸调pH＝1～2，用乙酸乙酯30mL萃取后，有机相， 减压浓缩得到化合物TM14。MS m/z：492.2[M+1]+.1H NMR(400MHz，CD3OD)δ＝8.47-8.38(m，1H)，8.31-8.21(m，1H)，8.16-8.06(m，1H)，7.55-7.48(m，1H)，7.47-7.15(m，5H)，7.14-7.06(m，2H)，5.31-5.05(m，2H)，2.86-2.74(m，1H)，2.64-2.54(m，1H)，2.46-2.37(m，1H)，1.21-1.07(m，3H).Compound TM14_9 (350.00 mg, 673.10 μmol) was added to tetrahydrofuran (6 mL), methanol (6 mL) and water (2 mL), and then lithium hydroxide (112.97 mg, 2.69 mmol) was added. The reaction solution was cooled to room temperature after stirring at 20°C for 3 hours. 20 mL of water was added to the reaction solution, and 10 mL of methyl tert-butyl ether was used for extraction. The organic phase was discarded, and the pH was adjusted to 1-2 with 1 M dilute hydrochloric acid. After extraction with 30 mL of ethyl acetate, the organic phase was The residue was concentrated under reduced pressure to give compound TM14. MS m/z: 492.2 [M+1] + . 1 H NMR (400 MHz, CD 3 OD) δ = 8.47-8.38 (m, 1H), 8.31-8.21 (m, 1H), 8.16-8.06 (m, 1H), 7.55-7.48 (m, 1H), 7.47-7.15 (m, 5H), 7.14-7.06 (m, 2H), 5.31-5.05 (m, 2H), 2.86-2.74 (m, 1H), 2.64-2.54 (m, 1H), 2.46-2.37 (m, 1H), 1.21-1.07 (m, 3H).

实施例14
Embodiment 14

步骤1：化合物TM15_2的合成Step 1: Synthesis of compound TM15_2

将化合物TM15_1(2g，19.78mmol)和对氟硝基苯(2.79g，19.78mmol)溶于2-甲基亚砜(20mL)中，加入碳酸钾(5.47g，39.55mmol)，置换氮气三次，80℃搅拌2hr。反应结束，加入50mL水淬灭反应，水相用乙酸乙酯萃取(30mL×3)，合并有机相，水洗(50mL×3)，无水硫酸钠干燥后。快速硅胶柱(石油醚∶乙酸乙酯＝100∶1＝90∶10)纯化。得到产物TM15_2。MSm/z：223.1[M+1]+.1H NMR(400MHz，CDCl3)δ(ppm)＝8.66(d，J＝4.8Hz，1H)，8.35-8.25(m，2H)，7.48-7.39(m，2H)，6.92-6.86(m，1H)Compound TM15_1 (2 g, 19.78 mmol) and p-fluoronitrobenzene (2.79 g, 19.78 mmol) were dissolved in 2-methyl sulfoxide (20 mL), potassium carbonate (5.47 g, 39.55 mmol) was added, nitrogen was replaced three times, and the mixture was stirred at 80°C for 2 hours. After the reaction was completed, 50 mL of water was added to quench the reaction, the aqueous phase was extracted with ethyl acetate (30 mL×3), the organic phases were combined, washed with water (50 mL×3), and dried over anhydrous sodium sulfate. Purification was performed on a rapid silica gel column (petroleum ether: ethyl acetate = 100:1 = 90:10). The product TM15_2 was obtained. MSm/z: 223.1[M+1] + .1H NMR (400MHz, CDCl 3 ) δ (ppm) = 8.66 (d, J = 4.8Hz, 1H), 8.35-8.25 (m, 2H), 7.48-7.39 (m, 2H), 6.92-6.86 (m, 1H)

步骤2：化合物TM15_3的合成Step 2: Synthesis of compound TM15_3

将TM15_2(3.8g，17.10mmol)溶于四氢呋喃中(16mL)，加入铁(3.82g，68.40mmol)，氯化铵(3.66g，68.40mmol)和水(12mL)，置换三次氮气后40℃搅拌48hr。反应液过滤，固体用乙酸乙酯洗涤(30mL)，分液，液体用乙酸乙酯萃取(30mL×3)，合并有机相并用无水硫酸钠干燥后减压旋干得TM15_3，MS m/z：193.1[M+1]+.1H NMR(400MHz，CDCl3)δ(ppm)＝8.53-8.49(m，1H)，7.06-7.01(m，2H)，6.74-6.68(m，3H)，3.77-3.51(m，2H).TM15_2 (3.8 g, 17.10 mmol) was dissolved in tetrahydrofuran (16 mL), iron (3.82 g, 68.40 mmol), ammonium chloride (3.66 g, 68.40 mmol) and water (12 mL) were added, and the nitrogen was replaced three times and stirred at 40°C for 48 hours. The reaction solution was filtered, the solid was washed with ethyl acetate (30 mL), separated, and the liquid was extracted with ethyl acetate (30 mL×3). The organic phases were combined and dried over anhydrous sodium sulfate and then dried under reduced pressure to obtain TM15_3, MS m/z: 193.1 [M+1] + . 1 H NMR (400 MHz, CDCl 3 )δ(ppm)＝8.53-8.49 (m, 1H), 7.06-7.01 (m, 2H), 6.74-6.68 (m, 3H), 3.77-3.51 (m, 2H).

步骤3：化合物TM15_4的合成Step 3: Synthesis of compound TM15_4

将化合物TM14_7(300.00mg，0.76mmol)和TM15_3(219.05mg，1.14mmol)溶于特戊酸(2mL)中，130℃搅拌5hr。反应结束，加入20mL水淬灭反应，水相用乙酸乙酯萃取(20mL×3)，合并有机相，水洗(20mL×3)，无水硫酸钠干燥后拌样。快速硅胶柱(石油醚∶乙酸乙酯＝100∶1-60∶40)纯化。得到产物TM15_4。MS m/z：525.0[M+1]+.1H NMR(400MHz，CDCl3)δ(ppm)＝8.56(d，J＝4.8Hz，1H)，7.74-7.72(m，1H)，7.38-7.31(m，4H)，7.24-7.21(m，2H)，7.09-7.07(m，1H)，6.87-6.84(m，2H)，6.79-6.76(m，1H)，5.05-4.90(m，2H)，4.09-3.97(m，2H)，2.81-2.71(m，1H)，2.50-2.38(m，2H)，1.20-1.15(m，6H).Compound TM14_7 (300.00 mg, 0.76 mmol) and TM15_3 (219.05 mg, 1.14 mmol) were dissolved in pivalic acid (2 mL) and stirred at 130°C for 5 hr. After the reaction was completed, 20 mL of water was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with water (20 mL×3), dried over anhydrous sodium sulfate, and then mixed. Purification was performed on a rapid silica gel column (petroleum ether: ethyl acetate = 100:1-60:40). The product TM15_4 was obtained. MS m/z: 525.0[M+1] + . 1 H NMR (400MHz, CDCl 3 ) δ (ppm) = 8.56 (d, J = 4.8Hz, 1H), 7.74-7.72 (m, 1H), 7.38-7.31 (m, 4H), 7.24-7.21 (m, 2H), 7.09-7.07 (m, 1H), 6 .87-6.84(m, 2H), 6.79-6.76(m, 1H), 5.05-4.90(m, 2H), 4.09-3.97(m, 2H), 2.81-2.71(m, 1H), 2.50-2.38(m, 2H), 1.20-1.15(m, 6H).

步骤4：化合物TM15的合成Step 4: Synthesis of compound TM15

将TM15_4(0.4g，0.76mmol)和溶于四氢呋喃(4mL)中，搅拌下加入溶解在水(1mL)中的氢氧化锂(159.84mg，3.81mmol)，然后在25℃搅拌16hr反应结束，加入10mL水，水相用乙酸乙酯萃取(10mL×3)，弃去有机相，水相用2M盐酸调节pH＝2～3，用乙酸乙酯萃取(20mL×3)，合并有机相，无水硫酸钠干燥后减压旋干。浓缩物用乙腈溶解后加入去离子水冻干。得到化合物TM15，MS m/z：497.0[M+1]+.1H NMR(400MHz，DMSO-d6)δ(ppm)＝12.17-12.02(m，1H)，9.03-8.97(m，1H)，8.76-8.61(m，1H)，7.68-7.51(m，1H)，7.49-7.23(m，5H)，7.21-7.07(m，2H)，6.99-6.61(m，2H)，5.34-4.91(m，2H)，3.52-3.21(m，1H)，2.78-2.58(m，1H)，2.26-2.14(m，1H)，1.08-0.95(m，3H). TM15_4 (0.4 g, 0.76 mmol) was dissolved in tetrahydrofuran (4 mL), and lithium hydroxide (159.84 mg, 3.81 mmol) dissolved in water (1 mL) was added under stirring, and then stirred at 25°C for 16 hours until the reaction was completed, 10 mL of water was added, the aqueous phase was extracted with ethyl acetate (10 mL×3), the organic phase was discarded, the aqueous phase was adjusted to pH=2-3 with 2M hydrochloric acid, extracted with ethyl acetate (20 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, and then dried under reduced pressure. The concentrate was dissolved in acetonitrile and added with deionized water for freeze drying. Compound TM15 was obtained, MS m/z: 497.0 [M+1] + . 1 H NMR (400 MHz, DMSO-d 6 ) δ (ppm) = 12.17-12.02 (m, 1H), 9.03-8.97 (m, 1H), 8.76-8.61 (m, 1H), 7.68-7.51 (m, 1H), 7.49-7.23 (m, 5H), 7.21-7.07 (m, 2H), 6.99-6.61 (m, 2H), 5.34-4.91 (m, 2H), 3.52-3.21 (m, 1H), 2.78-2.58 (m, 1H), 2.26-2.14 (m, 1H), 1.08-0.95 (m, 3H).

实施例15
Embodiment 15

步骤1：化合物TM16_1的合成Step 1: Synthesis of compound TM16_1

将TM01(75mg，152.77μmol)溶解于二氯甲烷(3mL)中，将反应体系降温至0℃，加入草酰氯(29.09mg，229.15μmol)，滴加时反应温度不超过10℃，然后加一滴N，N-二甲基甲酰胺，滴加完毕，反应体系20℃搅拌2hr。反应结束，减压浓缩除去溶剂。得到TM16_1，粗品直接投下一步。MS m/z：505.2[M+1]+。TM01 (75 mg, 152.77 μmol) was dissolved in dichloromethane (3 mL), the reaction system was cooled to 0°C, oxalyl chloride (29.09 mg, 229.15 μmol) was added, and the reaction temperature did not exceed 10°C during the addition. Then a drop of N,N-dimethylformamide was added, and the reaction system was stirred at 20°C for 2 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. TM16_1 was obtained, and the crude product was directly used for the next step. MS m/z: 505.2[M+1] + .

步骤2：化合物TM16的合成Step 2: Synthesis of compound TM16

将TM16_1(17.44mg，183.33μmol)溶于二氯甲烷(0.5mL)，加入N，N-二异丙基乙胺(59.23mg，458.32μmol)，将反应体系降至0℃，滴入溶于二氯甲烷(0.5mL)的甲基磺酰胺(77.82mg，152.77μmol)，慢慢升温到20℃搅拌12hr。反应结束，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：ACN％＝20％-54％，8.0min)纯化得到TM16。MS m/z：568.1[M+1]+。1H NMR(400MHz，DMSO-d6)δppm 11.80-11.89(m，1H)8.13-8.17(m，1H)8.12(s，1H)7.82-7.92(m，1H)7.46(br s，4H)7.34-7.42(m，2H)7.11-7.19(m，3H)7.01-7.08(m，1H)5.49-5.68(m，2H)3.15-3.23(m，3H)2.74-2.82(m，1H)2.60(br dd，J＝13.63，7.63Hz，1H)2.36-2.45(m，1H)1.05-1.14(m，3H)。TM16_1 (17.44 mg, 183.33 μmol) was dissolved in dichloromethane (0.5 mL), N, N-diisopropylethylamine (59.23 mg, 458.32 μmol) was added, the reaction system was cooled to 0°C, methylsulfonamide (77.82 mg, 152.77 μmol) dissolved in dichloromethane (0.5 mL) was added dropwise, the temperature was slowly raised to 20°C and stirred for 12 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure. TM16 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; flowability: [water (0.04% HCl)-ACN]; gradient: ACN% = 20%-54%, 8.0 min). MS m/z: 568.1[M+1] + . 1 H NMR (400MHz, DMSO-d 6 ) δppm 11.80-11.89 (m, 1H) 8.13-8.17 (m, 1H) 8.12 (s, 1H) 7.82-7.92 (m, 1H) 7.46 (br s, 4H) 7.34-7.42 (m, 2H) 7.11-7.19 (m, 3H) 7. 01-7.08 (m, 1H) 5.49-5.68 (m, 2H) 3.15-3.23 (m, 3H) 2.74-2.82 (m, 1H) 2.60 (br dd, J=13.63, 7.63Hz, 1H) 2.36-2.45 (m, 1H) 1.05-1.14 (m, 3H).

实施例16
Example 16

步骤1：化合物TM17的合成Step 1: Synthesis of compound TM17

将化合物TM08(100mg，198.03μmol)溶于二氯甲烷(3mL)，加入N，N-二异丙基乙胺(76.78mg，594.10μmol，103.48μL)和N-(3-二甲基氨丙基)-N-乙基碳二亚胺(56.94mg，297.05μmol)，20℃搅拌15min后，加入甲基磺酰胺(56.51mg，594.10μmol)和4-二甲氨基吡啶(29.03mg，237.64μmol)，20℃搅拌12hr。反应结束，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：ACN％＝20％-60％，8.0min)纯化得到TM17。MSm/z：582.1[M+1]+。1H NMR(400MHz，DMSO-d6)δppm 11.28-11.52(m，1H)8.09-8.23(m，1H)7.91-8.03(m，1H)7.81-7.91(m，1H)7.48(br s，4H)7.39(br s，2H)7.16(br s，3H)6.97-7.09(m，1H)5.45-5.68(m，2H)3.20(br s，3H)2.66(br s，2H)1.14(br s，6H).Compound TM08 (100 mg, 198.03 μmol) was dissolved in dichloromethane (3 mL), and N,N-diisopropylethylamine (76.78 mg, 594.10 μmol, 103.48 μL) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide (56.94 mg, 297.05 μmol) were added. After stirring at 20° C. for 15 min, methylsulfonamide (56.51 mg, 594.10 μmol) and 4-dimethylaminopyridine (29.03 mg, 237.64 μmol) were added, and stirred at 20° C. for 12 hr. After the reaction was completed, the solvent was removed by concentration under reduced pressure. TM17 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30 mm*3 μm; flowability: [water (0.04% HCl)-ACN]; gradient: ACN%=20%-60%, 8.0 min). MS m/z: 582.1 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 11.28-11.52 (m, 1H) 8.09-8.23 (m, 1H) 7.91-8.03 (m, 1H) 7.81-7.91 (m, 1H) 7.48 (br s, 4H) 7.39 (br s, 2H) 7.16 (br s, 3H) 6.97 -7.09(m,1H)5.45-5.68(m,2H)3.20(br s,3H)2.66(br s,2H)1.14(br s,6H).

实施例17
Embodiment 17

步骤1：化合物TM18的合成 Step 1: Synthesis of compound TM18

将化合物TM03(50.00mg，101.85μmol)溶于二氯甲烷(2mL)，加入N，N-二异丙基乙胺(32.91mg，254.61μmol，44.35μL)和N-(3-二甲基氨丙基)-N-乙基碳二亚胺(23.43mg，122.22μmol)，20℃搅拌15min后，加入甲基磺酰胺(29.06mg，305.54μmol)和4-二甲氨基吡啶(14.93mg，122.22μmol)，20℃搅拌12hr。反应结束，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：(ACN％＝20％-52％，8.0min)纯化得到TM18。MS m/z：568.0[M+1]+。1H NMR(400MHz，METHANOL-d4)δppm 8.29-8.34(m，1H)8.11-8.19(m，1H)7.92-7.96(m，1H)7.50(d，J＝8.66Hz，4H)7.36-7.43(m，3H)7.34(d，J＝8.91Hz，2H)7.14(d，J＝8.66Hz，1H)5.40-5.45(m，2H)3.22(s，3H)2.80-2.89(m，1H)2.60-2.76(m，2H)1.19-1.27(m，3H)。Compound TM03 (50.00 mg, 101.85 μmol) was dissolved in dichloromethane (2 mL), and N,N-diisopropylethylamine (32.91 mg, 254.61 μmol, 44.35 μL) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide (23.43 mg, 122.22 μmol) were added. After stirring at 20° C. for 15 min, methylsulfonamide (29.06 mg, 305.54 μmol) and 4-dimethylaminopyridine (14.93 mg, 122.22 μmol) were added, and stirred at 20° C. for 12 hr. After the reaction was completed, the solvent was removed by concentration under reduced pressure. TM18 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; flowability: [water (0.04% HCl)-ACN]; gradient: (ACN%=20%-52%, 8.0 min). MS m/z: 568.0 [M+1] + . 1 H NMR (400 MHz, METHANOL-d4) δ ppm 8.29-8.34 (m, 1H) 8.11-8.19 (m, 1H) 7.92-7.96 (m, 1H) 7.50 (d, J = 8.66Hz, 4H) 7.36-7.43 (m, 3H) 7.34 (d, J = 8.91Hz, 2H) 7.14 (d, J = 8.66Hz, 1H) 5.40-5. 45(m,2H)3.22(s,3H)2.80-2.89(m,1H)2.60-2.76(m,2H)1.19-1.27(m,3H).

实施例18
Embodiment 18

步骤1：化合物TM19的合成Step 1: Synthesis of compound TM19

将化合物TM02(50.00mg，101.85μmol)溶于二氯甲烷(2mL)，加入N，N-二异丙基乙胺(32.91mg，254.61μmol)和N-(3-二甲基氨丙基)-N-乙基碳二亚胺(23.43mg，122.22μmol)，20℃搅拌15min后，加入甲基磺酰胺(29.06mg，305.54μmol)和4-二甲氨基吡啶(14.93mg，122.22μmol)，20℃搅拌12hr。反应结束，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：(ACN％＝20％-52％，8.0min)纯化得到TM19。MSm/z：568.0[M+1]+。1H NMR(400MHz，DMSO-d6)δppm 11.74-11.83(m，1H)8.18(s，1H)7.81-7.93(m，2H)7.44-7.51(m，2H)7.38(br d，J＝8.17Hz，2H)7.31(br d，J＝3.96Hz，2H)7.13(br d，J＝7.67Hz，3H)7.04(d，J＝8.66Hz，1H)5.29-5.47(m，2H)3.19(s，3H)2.71-2.79(m，1H)2.53-2.59(m，1H)2.31-2.40(m，1H)1.04-1.11(m，3H)。Compound TM02 (50.00 mg, 101.85 μmol) was dissolved in dichloromethane (2 mL), N, N-diisopropylethylamine (32.91 mg, 254.61 μmol) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide (23.43 mg, 122.22 μmol) were added, and stirred at 20° C. for 15 min. Methanesulfonamide (29.06 mg, 305.54 μmol) and 4-dimethylaminopyridine (14.93 mg, 122.22 μmol) were added, and stirred at 20° C. for 12 hr. After the reaction was completed, the solvent was removed by concentration under reduced pressure. TM19 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobility: [water (0.04% HCl)-ACN]; gradient: (ACN%=20%-52%, 8.0min). MS m/z: 568.0 [M+1] + . 1H NMR (400MHz, DMSO-d6) δ ppm 11.74-11.83 (m, 1H) 8.18 (s, 1H) 7.81-7.93 (m, 2H) 7.44-7.51 (m, 2H) 7.38 (br d, J=8.17 Hz, 2H) 7.31 (br d, J=3.96 Hz, 2H) 7.13 (br d, J=7.67Hz, 3H) 7.04 (d, J=8.66Hz, 1H) 5.29-5.47 (m, 2H) 3.19 (s, 3H) 2.71-2.79 (m, 1H) 2.53-2.59 (m, 1H) 2.31-2.40 (m, 1H) 1.04-1.11 (m, 3H).

实施例19
Embodiment 19

步骤1：化合物TM20_2的合成Step 1: Synthesis of compound TM20_2

将化合物TM14_7(60mg，151.93μmol)加到特戊酸(0.3mL)中，然后加入TM20_1(93.07mg，455.79μmol)，130℃搅拌5hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(5mL×3)萃取，合并有机相，用饱和食盐水(10mL)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝1∶1)，得到化合物TM20_2，MS m/z：537.0[M+1]+。Compound TM14_7 (60 mg, 151.93 μmol) was added to pivalic acid (0.3 mL), and then TM20_1 (93.07 mg, 455.79 μmol) was added, and stirred at 130°C for 5 hr. The reaction system was cooled to room temperature, and diluted with saturated sodium bicarbonate solution (5 mL), extracted with ethyl acetate (5 mL×3), and the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:1) to obtain compound TM20_2, MS m/z: 537.0[M+1] + .

步骤2：化合物TM20的合成Step 2: Synthesis of compound TM20

将TM20_2(60.00mg，111.74μmol)加到四氢呋喃(0.6mL)、甲醇(0.6mL)和水(0.2mL)中，加入一水合氢氧化锂(8.03mg，335.21μmol)，45℃搅拌12hr。反应结束，将反应液降至室温，用稀盐酸(3M)调pH＝6～7，加乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：ACN％＝10％-56％，8.0min)纯化得到TM20。MS m/z：509.0[M+1]+。1H NMR(400MHz，CD3OD)δppm 8.04-8.09(m，1H)7.82-7.89(m，2H)7.44-7.50(m，2H)7.35(d，J＝8.41Hz，2H)7.24-7.32(m，2H)7.05-7.15(m，3H)5.29-5.36(m，2H)2.76-2.84(m，1H)2.66-2.74(m，1H)2.48-2.56(m，1H)1.17-1.24(m，3H). TM20_2 (60.00 mg, 111.74 μmol) was added to tetrahydrofuran (0.6 mL), methanol (0.6 mL) and water (0.2 mL), and lithium hydroxide monohydrate (8.03 mg, 335.21 μmol) was added, and stirred at 45°C for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (3 M), and ethyl acetate (10 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. TM20 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30 mm*3 μm; flowability: [water (0.04% HCl)-ACN]; gradient: ACN%=10%-56%, 8.0 min). MS m/z: 509.0[M+1] + . 1 H NMR (400MHz, CD 3 OD) δppm 8.04-8.09 (m, 1H) 7.82-7.89 (m, 2H) 7.44-7.50 (m, 2H) 7.35 (d, J=8.41Hz, 2H) 7.24-7.32 (m, 2H) 7.05-7.15 (m, 3H) 5.29-5. 36(m,2H)2.76-2.84(m,1H)2.66-2.74(m,1H)2.48-2.56(m,1H)1.17-1.24(m,3H).

实施例20
Embodiment 20

步骤1：化合物TM21_2的合成Step 1: Synthesis of compound TM21_2

将TM21_2(55g，274.71mmol)加入到四氢呋喃(300mL)和水(300mL)中，再加入氢氧化锂(34.58g，824.12mmol)，25℃搅拌12hr，减压浓缩除去溶剂，再用乙酸乙酯(1000mL)萃取，收集水相后，减压浓缩除去溶剂，再用稀盐酸(3M)调节pH至3～4，过滤，收集滤饼，减压浓缩除去溶剂。得到TM21_2，1H NMR(400MHz，DMSO-d6)δ＝8.53(s，1H)，2.56(s，3H).TM21_2 (55 g, 274.71 mmol) was added to tetrahydrofuran (300 mL) and water (300 mL), and then lithium hydroxide (34.58 g, 824.12 mmol) was added, and the mixture was stirred at 25°C for 12 hours, and the solvent was removed by concentrating under reduced pressure, and then extracted with ethyl acetate (1000 mL). After collecting the aqueous phase, the solvent was removed by concentrating under reduced pressure, and the pH was adjusted to 3-4 with dilute hydrochloric acid (3 M), and the mixture was filtered, the filter cake was collected, and the solvent was removed by concentrating under reduced pressure. TM21_2 was obtained, 1 H NMR (400 MHz, DMSO-d 6 ) δ=8.53 (s, 1H), 2.56 (s, 3H).

步骤2：化合物TM21_3的合成Step 2: Synthesis of compound TM21_3

将TM21_2(10.00g，53.71mmol)加入甲苯(67mL)中，置换氮气3次，加入三乙胺(16.30g，161.13mmol)和叠氮磷酸二苯酯(33.26g，120.85mmol)，45℃搅拌1.5hr，加入叔丁醇(67mL)，升温到115℃，搅拌4hr。减压浓缩除去溶剂，经自动过柱机纯化(淋洗液：石油醚∶乙酸乙酯＝1∶0，1∶0至0∶1).得到TM21_3Ms m/z＝258.1[M+H]+，直接用于下一步。TM21_2 (10.00 g, 53.71 mmol) was added to toluene (67 mL), nitrogen was replaced 3 times, triethylamine (16.30 g, 161.13 mmol) and diphenylphosphoryl azide (33.26 g, 120.85 mmol) were added, stirred at 45°C for 1.5 hr, tert-butyl alcohol (67 mL) was added, the temperature was raised to 115°C, and stirred for 4 hr. The solvent was removed by concentration under reduced pressure, and the mixture was purified by automatic column chromatography (eluent: petroleum ether: ethyl acetate = 1:0, 1:0 to 0:1). TM21_3 Ms m/z = 258.1 [M+H] + was obtained, which was used directly in the next step.

步骤3：化合物TM21_4的合成Step 3: Synthesis of compound TM21_4

将TM21_3(5g，19.43mmol)溶于二氯甲烷(80mL)，然后依次加入对氯苄溴(3.99g，19.43mmol)和二异丙基乙胺(5.02g，38.86mmol)，反应溶液在室温(15℃)下搅拌12hr。反应结束后向反应溶液中加入饱和氯化铵水溶液(50mL)和二氯甲烷(50mL)分离有机相，有机相减压浓缩，得到粗产品。经自动过柱机纯化(淋洗液：石油醚∶乙酸乙酯＝1∶0，1∶0至0∶1)得到化合物TM21_4.Msm/z＝382.1[M+H]+.1H NMR(400MHz，DMSO-d6)δ＝8.27(s，1H)，7.73(s，1H)，7.48(d，J＝8.4Hz，2H)，7.28(d，J＝8.4Hz，2H)，5.27(s，2H)，2.48(s，3H)，1.44(s，9H).TM21_3 (5 g, 19.43 mmol) was dissolved in dichloromethane (80 mL), and then p-chlorobenzyl bromide (3.99 g, 19.43 mmol) and diisopropylethylamine (5.02 g, 38.86 mmol) were added in sequence, and the reaction solution was stirred at room temperature (15°C) for 12 hours. After the reaction was completed, saturated aqueous ammonium chloride solution (50 mL) and dichloromethane (50 mL) were added to the reaction solution to separate the organic phase, and the organic phase was concentrated under reduced pressure to obtain a crude product. The compound TM21_4 was purified by automatic column chromatography (eluent: petroleum ether: ethyl acetate = 1:0, 1:0 to 0:1). Msm/z = 382.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ = 8.27 (s, 1H), 7.73 (s, 1H), 7.48 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 5.27 (s, 2H), 2.48 (s, 3H), 1.44 (s, 9H).

步骤4：化合物TM21_5的合成Step 4: Synthesis of compound TM21_5

将TM21_4(1g，2.62mmol)溶于四氢呋喃(5mL)，降温至0℃，然后加入钠氢(314.24mg，7.86mmol，60％纯度)和溴乙酸甲酯(440.65mg，2.88mmol)，反应溶液升温至室温20℃，搅拌2hr。反应结束后，向反应溶液中滴加饱和氯化铵水溶液(20mL)淬灭反应，然后用乙酸乙酯(30mL×2)萃取，分离有机相，有机相减压浓缩得到粗品。粗品通过自动过柱机(淋洗液：石油醚∶乙酸乙酯＝1∶0，1∶0至1∶3)，纯化得到产物化合物TM21_5，直接用于下一步。TM21_4 (1 g, 2.62 mmol) was dissolved in tetrahydrofuran (5 mL), cooled to 0°C, and then sodium hydrogen (314.24 mg, 7.86 mmol, 60% purity) and methyl bromoacetate (440.65 mg, 2.88 mmol) were added. The reaction solution was heated to room temperature 20°C and stirred for 2 hours. After the reaction was completed, saturated aqueous ammonium chloride solution (20 mL) was added dropwise to the reaction solution to quench the reaction, and then extracted with ethyl acetate (30 mL×2), the organic phase was separated, and the organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was purified by an automatic column machine (eluent: petroleum ether: ethyl acetate = 1:0, 1:0 to 1:3) to obtain the product compound TM21_5, which was directly used in the next step.

步骤5：化合物TM21_6的合成Step 5: Synthesis of compound TM21_6

将TM21_5(430mg，947.26μmol)加到特戊酸(0.7mL)中，然后加入TM01_5A(264.58mg，1.42mmol)，130℃搅拌2hr。反应结束，缓慢加入5mL饱和碳酸氢钠溶液淬灭反应，乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤并减压旋干，粗品通过自动过柱机(淋洗液：石油醚∶乙酸乙酯＝1∶0，1∶0至1∶3)纯化得到TM21_6，直接用于下一步。Ms m/z＝592.1[M+H]+.TM21_5 (430 mg, 947.26 μmol) was added to pivalic acid (0.7 mL), and then TM01_5A (264.58 mg, 1.42 mmol) was added, and stirred at 130°C for 2 hr. After the reaction was completed, 5 mL of saturated sodium bicarbonate solution was slowly added to quench the reaction, and ethyl acetate (10 mL×3) was used for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure. The crude product was purified by an automatic column machine (eluent: petroleum ether: ethyl acetate = 1:0, 1:0 to 1:3) to obtain TM21_6, which was used directly in the next step. Ms m/z = 592.1 [M+H] + .

步骤6：化合物TM21的合成 Step 6: Synthesis of compound TM21

将TM21_6(290mg，489.83μmol)溶于盐酸/乙酸乙酯(4M，5mL)室温15℃，搅拌0.5hr。反应结束后，将反应溶液减压浓缩。得到化合物TM21。Ms m/z＝492.1[M+H]+.1H NMR(400MHz，DMSO-d6)δ＝10.33(s，1H)，8.15(d，J＝4.8Hz，1H)，7.86(t，J＝7.6Hz，1H)，7.51-7.38(m，5H)，7.35(d，J＝8.8Hz，2H)，7.26-7.19(m，1H)，7.16-7.11(m，3H)，7.04(d，J＝8.4Hz，1H)，5.47(s，2H)，3.87(s，2H)，3.64(s，3H).TM21_6 (290 mg, 489.83 μmol) was dissolved in hydrochloric acid/ethyl acetate (4 M, 5 mL) at room temperature of 15°C and stirred for 0.5 hr. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain compound TM21. Ms m/z=492.1[M+H] + . 1 H NMR (400MHz, DMSO-d 6 ) δ=10.33 (s, 1H), 8.15 (d, J=4.8Hz, 1H), 7.86 (t, J=7.6Hz, 1H), 7.51-7.38 (m, 5H), 7.35 (d, J=8.8Hz, 2H), 7. 26-7.19 (m, 1H), 7.16-7.11 (m, 3H), 7.04 (d, J=8.4Hz, 1H), 5.47 (s, 2H), 3.87 (s, 2H), 3.64 (s, 3H).

实施例21
Embodiment 21

步骤1：化合物TM22_1和化合物TM23_1的合成Step 1: Synthesis of Compound TM22_1 and Compound TM23_1

化合物TM04_2通过制备SFC(色谱柱：DAICELCHIRALCELOJ(250mm*30mm，10μm)；流动相：[A：CO2-B：MeOH(0.1％NH3H2O)]；B％：60％，等压洗脱模式)，分离时间：3mins，分离得到化合物TM22_1(保留时间：1.441min，ee值为100％)。和化合物TM23_1(保留时间：1.786min，ee值为99.76％)Compound TM04_2 was separated by preparative SFC (chromatographic column: DAICELCHIRALCELOJ (250mm*30mm, 10μm); mobile phase: [A: CO 2 -B: MeOH (0.1% NH 3 H 2 O)]; B%: 60%, isobaric elution mode), separation time: 3mins, to obtain compound TM22_1 (retention time: 1.441min, ee value 100%) and compound TM23_1 (retention time: 1.786min, ee value 99.76%).

SFC分析方法：色谱柱：Chiralpak OJ-3，50×4.6mm I.D.，3μm，流动相：A：CO2B：MeOH[0.2％NH3(7M in MeOH)，v/v)，梯度：A∶B＝95∶5流速：3.4mL/min，柱温：35℃，分离时间：3mins。SFC analysis method: Chromatographic column: Chiralpak OJ-3, 50×4.6 mm ID, 3 μm, mobile phase: A: CO 2 B: MeOH [0.2% NH 3 (7M in MeOH), v/v), gradient: A: B=95:5, flow rate: 3.4 mL/min, column temperature: 35° C., separation time: 3 mins.

步骤2：化合物TM22的合成Step 2: Synthesis of compound TM22

将TM22_1(100.00mg，186.23μmol)溶于四氢呋喃(0.5mL)和水中(0.5mL)，再加入氢氧化锂(11.72mg，279.34μmol)，20℃搅拌2.5hr。反应结束，将反应液用1M的稀盐酸调pH至3～4，待析出固体，过滤收集滤饼减压浓缩除去溶剂。通过制备HPLC(色谱柱：Phenomenex Luna C18 75*30mm*3μm；流动相：[H2O(0.04％HCl)-ACN]；梯度：20％-65％ACN，8.0min)纯化，冻干后得到TM22。MSm/z：509.1[M+1]+。1H NMR(400MHz，DMSO-d6)：δ＝7.96-8.05(m，1H)，7.67-7.75(m，1H)，7.47(d，J＝8.5Hz，2H)，7.19-7.41(m，4H)，7.12(br d，J＝6.5Hz，2H)，6.84-6.92(m，2H)，5.08-5.45(m，2H)，2.64-2.70(m，1H)，2.56-2.50(m，1H)，2.23(br dd，J＝14.4，6.8Hz，1H)，1.04ppm(d，J＝7.0Hz，3H).TM22_1 (100.00 mg, 186.23 μmol) was dissolved in tetrahydrofuran (0.5 mL) and water (0.5 mL), and then lithium hydroxide (11.72 mg, 279.34 μmol) was added and stirred at 20°C for 2.5 hr. After the reaction was completed, the reaction solution was adjusted to pH 3-4 with 1M dilute hydrochloric acid, and the solid was precipitated, the filter cake was collected by filtration, and the solvent was removed by vacuum concentration. TM22 was purified by preparative HPLC (chromatographic column: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [H 2 O (0.04% HCl)-ACN]; gradient: 20%-65% ACN, 8.0 min) and freeze-dried to obtain TM22. MSm/z: 509.1[M+1] + . 1H NMR (400MHz, DMSO-d6): δ=7.96-8.05 (m, 1H), 7.67-7.75 (m, 1H), 7.47 (d, J=8.5Hz, 2H), 7.19-7.41 (m, 4H), 7.12 (br d, J=6.5Hz, 2H), 6.84-6.92 (m, 2H), 5 .08-5.45 (m, 2H), 2.64-2.70 (m, 1H), 2.56-2.50 (m, 1H), 2.23 (br dd, J=14.4, 6.8Hz, 1H), 1.04ppm (d, J=7.0Hz, 3H).

步骤3：化合物TM23的合成Step 3: Synthesis of compound TM23

将TM23_1(100.00mg，186.23μmol)溶于四氢呋喃(0.5mL)和水中(0.5mL)，再加入氢氧化锂(11.72mg，279.34μmol)，20℃搅拌2.5hr。反应结束，将反应液pH调至3～4，待析出固体，过滤收集滤饼减压浓缩除去溶剂。通过制备HPLC(色谱柱：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：20％-65％ACN，8.0min)纯化，冻干后得到TM23。MS m/z：509.1[M+1]+。1H NMR(400MHz，DMSO-d6)：δ＝7.95-8.05(m，1H)，7.51-7.56(m，1H)，7.39-7.49(m，4H)，7.25-7.35(m，2H)，7.00-7.13(m，2H)，6.78-6.90(m，2H)，4.95-5.37(m，2H)，2.58-2.70(m，1H)，2.57-2.54(m，1H)，2.28-2.36(m，1H)，0.98ppm(br d，3H).TM23_1 (100.00 mg, 186.23 μmol) was dissolved in tetrahydrofuran (0.5 mL) and water (0.5 mL), and then lithium hydroxide (11.72 mg, 279.34 μmol) was added and stirred at 20°C for 2.5 hr. After the reaction was completed, the pH of the reaction solution was adjusted to 3-4, and the solid was precipitated, and the filter cake was collected by filtration and concentrated under reduced pressure to remove the solvent. TM23 was purified by preparative HPLC (chromatographic column: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: 20%-65% ACN, 8.0 min) and freeze-dried to obtain TM23. MS m/z: 509.1[M+1] + . 1 H NMR (400MHz, DMSO-d6): δ = 7.95-8.05 (m, 1H), 7.51-7.56 (m, 1H), 7.39-7.49 (m, 4H), 7.25-7.35 (m, 2H), 7.00-7.13 (m, 2H), 6.78-6.90 (m, 2H), 4.95-5 .37(m, 2H), 2.58-2.70(m, 1H), 2.57-2.54(m, 1H), 2.28-2.36(m, 1H), 0.98ppm(br d, 3H).

实施例22
Embodiment 22

步骤1：化合物TM24_1的合成Step 1: Synthesis of compound TM24_1

在一个预先干燥的单口瓶中加入化合物TM14_8(1g，2.53mmol)，特戊酸(3.88g，37.98mmol，4.36mL)和化合物TM01_5(568.82mg，3.04mmol)混合后，在130℃下反应5hr。反应完成后，反应结束后，加入饱和碳酸氢钠水溶液(100mL)淬灭，pH调至7～8，用乙酸乙酯(100mL*3)萃取，合并有机相，有机相用饱和食盐水200mL洗涤，无水硫酸钠干燥，过滤，减压浓缩得到残余物。向残余物中加入70mL(MTBE∶EA＝10∶1)搅拌1h后，过滤收集滤饼，干燥，得到化合物TM24_1，直接用于下一步。MS m/z：520.1[M+1]+。Compound TM14_8 (1 g, 2.53 mmol), pivalic acid (3.88 g, 37.98 mmol, 4.36 mL) and compound TM01_5 (568.82 mg, 3.04 mmol) were added to a pre-dried single-mouth bottle and mixed, and reacted at 130°C for 5 hours. After the reaction was completed, saturated sodium bicarbonate aqueous solution (100 mL) was added to quench, the pH was adjusted to 7-8, and extracted with ethyl acetate (100 mL*3), the organic phases were combined, washed with saturated brine 200 mL, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. 70 mL (MTBE: EA = 10: 1) was added to the residue and stirred for 1 hour, and the filter cake was collected by filtration and dried to obtain compound TM24_1, which was directly used in the next step. MS m/z: 520.1 [M+1] + .

步骤2：化合物TM24的合成Step 2: Synthesis of compound TM24

在一个预先干燥好的单口瓶中加入TM24_1(400mg，769.26μmol)和四氢呋喃(6mL)，甲醇(6mL)和水(2mL)，后加入氢氧化锂(129.12mg，3.08mmol)，20℃搅拌3hr。反应结束后，用1M稀盐酸调pH＝1～2，后减压浓缩得到残余物，残余物用3mL DMF溶解后过滤送制备分离，经制备柱(Phenomenex luna C18 100*40mm*5μm；流动相：[H2O(0.04％HCl)-ACN]；梯度：25％-65％ACN，8.0min)分离冻干得到TM24，MS m/z：492.1[M+1]+，1H NMR(400MHz，DMSO-d6)δ＝8.56-8.53(m，1H)，8.41-8.36(m，1H)，8.23-8.20(m，1H)，8.05(s，1H)，7.52-7.46(m，2H)，7.45-7.34(m，4H)，7.26-7.18(m，2H)，5.49(br s，2H)，2.71-2.59(m，2H)，2.39-2.26(m，1H)，1.10-1.03(m，3H)。TM24_1 (400 mg, 769.26 μmol), tetrahydrofuran (6 mL), methanol (6 mL) and water (2 mL) were added to a pre-dried single-mouth bottle, followed by lithium hydroxide (129.12 mg, 3.08 mmol), and stirred at 20°C for 3 hr. After the reaction, the pH was adjusted to 1-2 with 1 M dilute hydrochloric acid, and then the residue was concentrated under reduced pressure. The residue was dissolved with 3 mL DMF and filtered for preparative separation. TM24 was separated and freeze-dried on a preparative column (Phenomenex luna C18 100*40mm*5μm; mobile phase: [H 2 O (0.04% HCl)-ACN]; gradient: 25%-65% ACN, 8.0 min) to obtain TM24, MS m/z: 492.1 [M+1] + , 1 H NMR (400 MHz, DMSO-d 6 )δ=8.56-8.53(m,1H),8.41-8.36(m,1H),8.23-8.20(m,1H),8.05(s,1H),7.52-7.46(m,2H),7.45-7.34(m,4H),7.26-7.18(m,2H),5.49(br s,2H), 2.71-2.59 (m, 2H), 2.39-2.26 (m, 1H), 1.10-1.03 (m, 3H).

实施例23
Embodiment 23

步骤1：化合物TM25_1的合成Step 1: Synthesis of compound TM25_1

将化合物TM01_5(100mg，253.22μmol)加到特戊酸(0.5mL)中，然后加入化合物TM20_1(155.12mg，759.66μmol)，130℃搅拌5hr，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(5mL×3)萃取，合并有机相，用饱和食盐水(10mL)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝1∶1)，得到化合物TM25_1，直接用于下一步。MS m/z：537.0[M+1]+。Compound TM01_5 (100 mg, 253.22 μmol) was added to pivalic acid (0.5 mL), and then compound TM20_1 (155.12 mg, 759.66 μmol) was added, and stirred at 130°C for 5 hours. The reaction system was cooled to room temperature, and saturated sodium bicarbonate solution (5 mL) was added for dilution, and extracted with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:1) to obtain compound TM25_1, which was directly used in the next step. MS m/z: 537.0[M+1] + .

步骤2：化合物TM25的合成Step 2: Synthesis of compound TM25

将TM25_1(100mg，186.23μmol)加到四氢呋喃(0.6mL)、甲醇(0.6mL)和水(0.2mL)中，加入一水合氢氧化锂(13.38mg，558.68μmol)，45℃搅拌12hr。反应结束，将反应液降至室温，用稀盐酸(3M)调pH＝6～7，加乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：(ACN％＝30％-65％，8.0min)纯化得到TM25。MS m/z：509.0[M+1]+.1H NMR(400MHz，DMSO-d6)δppm 8.14(dd，J＝4.95，1.24Hz，1H)7.83-7.92(m，2H)7.46-7.52(m，3H)7.38-7.41(m，2H)7.24-7.31(m，1H)7.08-7.19(m，3H)5.24-5.37(m，2H)2.68-2.71(m，1H)2.60(br d，J＝12.99Hz，1H)2.25-2.33(m，1H)1.07(d，J＝6.93Hz，3H).TM25_1 (100 mg, 186.23 μmol) was added to tetrahydrofuran (0.6 mL), methanol (0.6 mL) and water (0.2 mL), and lithium hydroxide monohydrate (13.38 mg, 558.68 μmol) was added, and stirred at 45°C for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and the pH was adjusted to 6-7 with dilute hydrochloric acid (3 M), and ethyl acetate (10 mL×3) was added for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. TM25 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobility: [water (0.04% HCl)-ACN]; gradient: (ACN%=30%-65%, 8.0min). MS m/z: 509.0 [M+1] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.14 (dd, J=4.95, 1.24 Hz, 1H) 7.83-7.92 (m, 2H) 7.46-7.52 (m, 3H) 7.38-7.41 (m, 2H) 7.24-7.31 (m, 1H) 7.08-7.19 (m, 3H) 5.24-5.37 (m, 2H) 2.68-2.71 (m, 1H) 2.60 (br d, J=12.99Hz, 1H) 2.25-2.33 (m, 1H) 1.07 (d, J=6.93Hz, 3H).

实施例24
Embodiment 24

将化合物TM01(50mg，101.85μmol)溶于二氯甲烷(1mL)，加入N，N-二异丙基乙胺(32.91mg，254.61μmol，44.35μL)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(29.29mg，152.77μmol)，20℃搅拌15min后，加入盐酸甲胺(27.51mg，407.38μmol)和4-二甲氨基吡啶(14.93mg，122.22μmol)，20℃搅拌1hr。反应完毕，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18100*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝5％-40％，8.0min)纯化得到化合物TM26。MS m/z：504.1[M+1]+。1H NMR(400MHz，DMSO-d6)δppm 8.13-8.18(m，1H)8.05-8.10(m，1H)7.81-7.90(m，2H)7.46-7.51(m，2H)7.36-7.45(m，4H)7.12-7.18(m，3H)7.03-7.07(m，1H)5.50-5.69(m，2H)2.54(br d，J＝2.63Hz，2H)2.48(d，J＝4.50Hz，3H)2.31-2.40(m，1H)1.02(d，J＝6.38Hz，3H)。Compound TM01 (50 mg, 101.85 μmol) was dissolved in dichloromethane (1 mL), and N,N-diisopropylethylamine (32.91 mg, 254.61 μmol, 44.35 μL) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (29.29 mg, 152.77 μmol) were added. After stirring at 20°C for 15 min, methylamine hydrochloride (27.51 mg, 407.38 μmol) and 4-dimethylaminopyridine (14.93 mg, 122.22 μmol) were added, and stirred at 20°C for 1 hr. After the reaction was completed, the solvent was removed by concentration under reduced pressure. Compound TM26 was purified by preparative HPLC (column type: Phenomenex Luna C18100*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%=5%-40%, 8.0min). MS m/z: 504.1 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 8.13-8.18 (m, 1H) 8.05-8.10 (m, 1H) 7.81-7.90 (m, 2H) 7.46-7.51 (m, 2H) 7.36-7.45 (m, 4H) 7.12-7.18 (m, 3H) 7.03-7.07 (m, 1H) 5.50-5.69 (m, 2H) 2.54 (br d, J=2.63Hz, 2H) 2.48 (d, J=4.50Hz, 3H) 2.31-2.40 (m, 1H) 1.02 (d, J=6.38Hz, 3H).

实施例25
Embodiment 25

将化合物TM01(50mg，101.85μmol)溶于二氯甲烷(1mL)，加入N，N-二异丙基乙胺(32.91mg，254.61μmol，44.35μL)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(29.29mg，152.77μmol)，20℃搅拌15min后，加入2，2，2-三氟乙胺(40.35mg，407.38μmol)和4-二甲氨基吡啶(14.93mg，122.22μmol)，20℃搅拌12hr。反应完毕，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 100*30mm*3μm；流动相：[水(0.04％HCl)-ACN]；梯度：(ACN％＝10％-45％，8.0min)纯化得到化合物TM27。MSm/z：572.0[M+1]+。1H NMR(400MHz，DMSO-d6)δppm 8.45(s，1H)8.08-8.20(m，1H)7.79-7.89(m，1H)7.62-7.71(m，1H)7.42-7.52(m，2H)7.22-7.39(m，4H)7.09(br d，J＝9.16Hz，3H)7.01(br d，J＝8.41Hz，1H)5.21-5.36(m，2H)3.77(br d，J＝6.19Hz，2H)2.65-2.69(m，1H)2.43(br s，1H)2.27-2.33(m，1H)1.01(br d，J＝4.33Hz，3H)。Compound TM01 (50 mg, 101.85 μmol) was dissolved in dichloromethane (1 mL), and N,N-diisopropylethylamine (32.91 mg, 254.61 μmol, 44.35 μL) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (29.29 mg, 152.77 μmol) were added. After stirring at 20°C for 15 min, 2,2,2-trifluoroethylamine (40.35 mg, 407.38 μmol) and 4-dimethylaminopyridine (14.93 mg, 122.22 μmol) were added, and stirred at 20°C for 12 hr. After the reaction was completed, the solvent was removed by concentration under reduced pressure. Compound TM27 was purified by preparative HPLC (column type: Phenomenex Luna C18 100*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; gradient: (ACN%=10%-45%, 8.0min). MS m/z: 572.0 [M+1] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.45 (s, 1H) 8.08-8.20 (m, 1H) 7.79-7.89 (m, 1H) 7.62-7.71 (m, 1H) 7.42-7.52 (m, 2H) 7.22-7.39 (m, 4H) 7.09 (br d, J=9.16 Hz, 3H) 7.01 (br d, J=8.41Hz, 1H) 5.21-5.36 (m, 2H) 3.77 (br d, J=6.19Hz, 2H) 2.65-2.69 (m, 1H) 2.43 (br s, 1H) 2.27-2.33 (m, 1H) 1.01 (br d, J= 4.33Hz, 3H).

实施例26
Embodiment 26

将化合物TM01(200mg，407.38μmol)溶于二氯甲烷(3mL)，加入N，N-二异丙基乙胺(177.39μL)和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(117.14mg，611.08μmol)，20℃搅拌15min后，加入2，2-二氟乙胺(132.10mg，1.63mmol)和4-二甲氨基吡啶(59.72mg，488.86μmol)，20℃搅拌12hr。反应完毕，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动相：[水(0.04％HCl)-ACN]； 梯度：(ACN％＝20％-55％，8.0min)纯化得到化合物TM28。MS m/z：554.0[M+1]+。1H NMR(400MHz，DMSO-d6)δppm 8.27(t，J＝5.88Hz，1H)8.11-8.17(m，1H)7.91(s，1H)7.83-7.89(m，1H)7.45-7.51(m，2H)7.37-7.42(m，2H)7.33(br d，J＝8.54Hz，2H)7.10-7.17(m，3H)7.01-7.06(m，1H)5.71-6.07(m，1H)5.34-5.50(m，2H)3.15-3.29(m，2H)2.61-2.69(m，1H)2.54(s，1H)2.32-2.40(m，1H)1.03(d，J＝6.80Hz，3H)。Compound TM01 (200 mg, 407.38 μmol) was dissolved in dichloromethane (3 mL), and N, N-diisopropylethylamine (177.39 μL) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (117.14 mg, 611.08 μmol) were added. After stirring at 20°C for 15 min, 2, 2-difluoroethylamine (132.10 mg, 1.63 mmol) and 4-dimethylaminopyridine (59.72 mg, 488.86 μmol) were added, and stirred at 20°C for 12 hr. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The product was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.04% HCl)-ACN]; Gradient: (ACN% = 20%-55%, 8.0 min) to obtain compound TM28. MS m/z: 554.0 [M+1] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.27 (t, J = 5.88 Hz, 1H) 8.11-8.17 (m, 1H) 7.91 (s, 1H) 7.83-7.89 (m, 1H) 7.45-7.51 (m, 2H) 7.37-7.42 (m, 2H) 7.33 (br d, J=8.54Hz, 2H) 7.10-7.17 (m, 3H) 7.01-7.06 (m, 1H) 5.71-6.07 (m, 1H) 5.34-5.50 (m, 2H) 3.15-3.29 (m, 2H) 2.61-2.69 (m, 1H) 2.54 (s, 1H) 2.32-2.4 0(m,1H)1.03(d,J=6.80Hz,3H).

实施例27
Embodiment 27

步骤1：化合物TM29_2的合成Step 1: Synthesis of compound TM29_2

将化合物TM14_7(86mg，217.77μmol)加到特戊酸(1mL)中，然后加入化合物TM29_1(49.27mg，217.77μmol)，130℃搅拌12hr。反应结束，将反应体系降至室温，加入饱和碳酸氢钠溶液(5mL)稀释，用乙酸乙酯(5mL×3)萃取，合并有机相，用饱和食盐水(5mL×3)洗，无水硫酸钠干燥，过滤，滤液减压浓缩除去溶剂。粗品用硅胶柱层析法分离(石油醚/乙酸乙酯＝0∶1)，得到化合物TM29_2。MS m/z：559.1[M+1]+。Compound TM14_7 (86 mg, 217.77 μmol) was added to pivalic acid (1 mL), and then compound TM29_1 (49.27 mg, 217.77 μmol) was added, and stirred at 130°C for 12 hours. After the reaction was completed, the reaction system was cooled to room temperature, and saturated sodium bicarbonate solution (5 mL) was added for dilution, and extracted with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated brine (5 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=0:1) to obtain compound TM29_2. MS m/z: 559.1[M+1] + .

步骤2：化合物TM29的合成Step 2: Synthesis of compound TM29

将化合物TM29_2(79mg，141.32μmol)溶于四氢呋喃(0.6mL)、甲醇(0.6mL)和水(0.2mL)，加入氢氧化锂(13.54mg，565.28μmol)，45℃搅拌12hr。反应结束，将反应液降至室温，加水(10mL)稀释，用稀盐酸(3M)调pH＝6-7，加乙酸乙酯(10mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，减压浓缩除去溶剂。通过制备HPLC(柱型：Phenomenex Luna C18 75*30mm*3μm；流动性：[水(0.04％HCl)-ACN]；梯度：(ACN％＝10％-47％，8.0min)纯化得到化合物TM29。MS m/z：531.0[M+1]+。1H NMR(400MHz，DMSO-d6)δppm 8.41-8.52(m，2H)8.12-8.19(m，1H)7.93-8.03(m，1H)7.59-7.70(m，1H)7.37-7.53(m，6H)7.26-7.35(m，2H)5.42-5.65(m，2H)2.58-2.74(m，2H)2.26-2.38(m，1H)0.98-1.12(m，3H)。Compound TM29_2 (79 mg, 141.32 μmol) was dissolved in tetrahydrofuran (0.6 mL), methanol (0.6 mL) and water (0.2 mL), and lithium hydroxide (13.54 mg, 565.28 μmol) was added, and stirred at 45°C for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water (10 mL), adjusted to pH = 6-7 with dilute hydrochloric acid (3 M), extracted with ethyl acetate (10 mL×3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent. Compound TM29 was purified by preparative HPLC (column type: Phenomenex Luna C18 75*30mm*3μm; flowability: [water (0.04% HCl)-ACN]; gradient: (ACN%=10%-47%, 8.0 min). MS m/z: 531.0 [M+1] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.41-8.52(m,2H)8.12-8.19(m,1H)7.93-8.03(m,1H)7.59-7.70(m,1H)7.37-7.53(m,6H)7.26-7.35(m,2H)5.42-5.65(m,2H)2.58-2.74(m,2H)2 .26-2.38(m,1H)0.98-1.12(m,3H).

实施例28化合物TM03的A晶型的制备Example 28 Preparation of Crystal Form A of Compound TM03

取化合物TM03(500mg，1.02mmol)加到乙酸乙酯(100mL)中，80℃搅拌至溶清，然后自然降温至30℃，加入石油醚(200mL)，然后25℃搅拌12hr。25℃下抽滤，残留固体样品置于真空干燥箱中(30℃)干燥过夜，得化合物TM03的A晶型。Compound TM03 (500 mg, 1.02 mmol) was added to ethyl acetate (100 mL), stirred at 80°C until dissolved, then cooled naturally to 30°C, petroleum ether (200 mL) was added, and then stirred at 25°C for 12 hours. The mixture was filtered at 25°C, and the residual solid sample was placed in a vacuum drying oven (30°C) and dried overnight to obtain Form A of compound TM03.

实施例29：化合物TM03的B晶型的制备Example 29: Preparation of Form B of Compound TM03

取化合物TM03(12.11g，24.67mmol)加到甲醇(96mL)中，90℃搅拌至溶清，然后自然降温至25℃，搅拌12hr。25℃下抽滤，残留固体样品置于真空干燥箱中(30℃)干燥过夜，得化合物TM03的B晶型。Compound TM03 (12.11 g, 24.67 mmol) was added to methanol (96 mL), stirred at 90°C until dissolved, then cooled naturally to 25°C and stirred for 12 hours. The mixture was filtered at 25°C, and the residual solid sample was placed in a vacuum drying oven (30°C) and dried overnight to obtain Form B of compound TM03.

实施例30：化合物TM03的D晶型的制备Example 30: Preparation of Form D of Compound TM03

取化合物TM03(100mg，203.69μmol)用乙酸乙酯(200mL)溶清后旋干，将浓缩物加到正庚烷(2mL)中，25℃搅拌144hr。25℃搅拌144hr，25℃下抽滤，残留固体样品置于真空干燥箱中(30℃)干燥过夜，得化合物TM03的D晶型。Compound TM03 (100 mg, 203.69 μmol) was dissolved in ethyl acetate (200 mL) and then dried by rotation. The concentrate was added to n-heptane (2 mL) and stirred at 25° C. for 144 hr. Stirred at 25° C. for 144 hr, filtered at 25° C., and the residual solid sample was placed in a vacuum drying oven (30° C.) and dried overnight to obtain the D crystal form of compound TM03.

实施例31化合物TM03的B晶型的吸湿性研究Example 31 Study on the Hygroscopicity of Crystal Form B of Compound TM03

实验材料：Experimental Materials:

SMS DVS Advantage动态蒸汽吸附仪SMS DVS Advantage Dynamic Vapor Sorption Tester

实验方法：Experimental methods:

取式化合物TM03的B晶型10～15mg置于DVS样品盘内进行测试。10-15 mg of the Form B of the compound TM03 was placed in a DVS sample tray for testing.

实验结果：Experimental results:

式化合物TM03的B晶型的DVS谱图基本上如图12所示，△W＝0.0923％。 The DVS spectrum of the B-type of the compound of formula TM03 is basically as shown in Figure 12, ΔW = 0.0923%.

实验结论：化合物TM03的B晶型几乎无吸湿性，其XRPD在DVS实验前后无变化。Experimental conclusion: The B crystal form of compound TM03 is almost non-hygroscopic, and its XRPD does not change before and after the DVS experiment.

实施例32：化合物TM03的B晶型的固体预稳定性试验Example 32: Solid pre-stability test of Form B of compound TM03

依据《原料药与制剂稳定性试验指导原则》(中国药典2015版四部通则9001)，考察在本公开实施例化合物结晶在高温(60℃，敞口)，高湿(相对湿度75％，敞口)条件下的稳定性。According to the "Guidelines for Stability Testing of APIs and Preparations" (Chinese Pharmacopoeia 2015 Edition Part IV General Rules 9001), the stability of the crystals of the compounds in the embodiments of the present disclosure under high temperature (60°C, open) and high humidity (relative humidity 75%, open) conditions was investigated.

称取本公开实施例化合物结晶15mg，置于玻璃样品瓶的底部，摊成薄薄一层。高温及高湿条件下放置的样品用铝箔纸封瓶口，并在铝箔纸上扎些小孔，保证样品能与环境空气充分接触；不同条件下放置的样品于第10天，30天取样检测(XRPD)，并将检测结果与0天的初始检测结果进行比较。Weigh 15 mg of the crystals of the compound of the embodiment of the present disclosure and place them at the bottom of a glass sample bottle, spreading them into a thin layer. The sample placed under high temperature and high humidity conditions is sealed with aluminum foil, and small holes are punched in the aluminum foil to ensure that the sample can fully contact with the ambient air; the samples placed under different conditions are sampled and tested (XRPD) on the 10th day and 30th day, and the test results are compared with the initial test results on day 0.

结论：化合物TM03的B晶型在高温、高湿条件下具有良好的稳定性。Conclusion: The B form of compound TM03 has good stability under high temperature and high humidity conditions.

生物数据Biological data

实验例1FLIPR钙流检测化合物对人源P2X3，人源P2X2/3通道信号的影响Experimental Example 1 Effects of FLIPR calcium flux detection compounds on human P2X3 and human P2X2/3 channel signals

实验目的：检测本发明化合物对人源P2X3受体和人源P2X2/3受体的抑制效应Experimental purpose: To detect the inhibitory effect of the compounds of the present invention on human P2X3 receptors and human P2X2/3 receptors

实验材料：Experimental Materials:

受试物：本发明化合物；对照品：αβ-methyleneATP，AF-219；溶媒：二甲基亚砜(DMSO)Test substance: compound of the present invention; Reference substance: αβ-methyleneATP, AF-219; Solvent: dimethyl sulfoxide (DMSO)

实验过程：Experimental process:

1.给药制剂储液配制方法1. Preparation method of drug delivery preparation stock solution

1.1.激动剂：称量适量的αβ-methylene ATP用无菌水配制成20mM的储液。分装后-80℃保存。1.1. Agonist: Weigh an appropriate amount of αβ-methylene ATP and prepare a 20mM stock solution with sterile water. Store at -80℃ after aliquoting.

1.2.抑制剂：称量适量的AF-219用DMSO配制成10mM的储液。分装后-20℃保存。1.2. Inhibitor: Weigh an appropriate amount of AF-219 and prepare a 10 mM stock solution with DMSO. Store at -20°C after aliquoting.

1.3.受试物：称量合适质量的受试物，根据公式：DMSO体积＝实际量×纯度/(分子量×理论浓度)，计算出所需的DMSO的体积，吸取相应体积的DMSO，之后将已经称量的受试物用吸取的DMSO溶解，配置20mM的储液，放置于-20℃保存。1.3. Test substance: Weigh a suitable mass of the test substance, calculate the required volume of DMSO according to the formula: DMSO volume = actual amount × purity / (molecular weight × theoretical concentration), aspirate the corresponding volume of DMSO, then dissolve the weighed test substance in the aspirated DMSO, prepare a 20mM stock solution, and store it at -20°C.

2.给药制剂工作液配制和化合物检测板准备2. Preparation of drug delivery solution and compound detection plate

2.1.工作液配制设计：在人源P2X3，P2X2/3受体测试之前，将激动剂及抑制剂储液和受试物储液取出。激动剂αβ-methylene ATP的检测浓度为300μM起始，3倍稀释，9个浓度，2个重复；抑制剂AF-219的检测浓度为10μM起始，3倍稀释，9个浓度，2个重复；受试物检测浓度为1μM或者100μM起始，3倍稀释，9个浓度，2个重复；受试物用αβ-methyleneATPEC80的浓度激活人源P2X3和P2X2/3通道。空白对照品为0.5％DMSO。2.1. Working solution preparation design: Before testing human P2X3 and P2X2/3 receptors, take out the agonist and inhibitor stock solutions and the test substance stock solutions. The detection concentration of the agonist αβ-methylene ATP is 300μM starting, 3-fold dilution, 9 concentrations, 2 replicates; the detection concentration of the inhibitor AF-219 is 10μM starting, 3-fold dilution, 9 concentrations, 2 replicates; the test substance detection concentration is 1μM or 100μM starting, 3-fold dilution, 9 concentrations, 2 replicates; the test substance activates human P2X3 and P2X2/3 channels with the concentration of αβ-methyleneATPEC80. The blank control is 0.5% DMSO.

2.2.化合物检测板准备：在ECHO555液体工作站上设置好打板程序。进行打板时，化合物板最终浓度需设置为检测浓度的3倍或者4倍。当进行化合物板准备时，ECHO专用LDV板需要首先使用溶剂DMSO或者双蒸水将化合物储存液稀释为10mM，1mM和0.1mM，进行化合物浓度梯度打板。进而再准备溶剂DMSO补液打板，使得每孔总体积补至200nL，保证DMSO含量每孔一致，此时化合物浓度为待测浓度600倍或者800倍。最后再使用排枪，补39.8μL缓冲液至化合物板每孔，使得DMSO在所有孔内的含量均为0.5％，并得到3倍或者4倍于待测浓度的化合物板。待测化合物板需当天进行打板，化合物板检测前，放置于室温氮气柜保存。2.2. Preparation of compound detection plate: Set up the plate-making program on the ECHO555 liquid workstation. When making the plate, the final concentration of the compound plate needs to be set to 3 or 4 times the detection concentration. When preparing the compound plate, the ECHO-specific LDV plate needs to first use the solvent DMSO or double distilled water to dilute the compound storage solution to 10mM, 1mM and 0.1mM, and make a compound concentration gradient plate. Then prepare the solvent DMSO to make the plate, so that the total volume of each well is replenished to 200nL, and ensure that the DMSO content is consistent in each well. At this time, the compound concentration is 600 times or 800 times the concentration to be tested. Finally, use a discharge gun to add 39.8μL of buffer to each well of the compound plate, so that the DMSO content in all wells is 0.5%, and a compound plate with a concentration of 3 or 4 times that to be tested is obtained. The compound plate to be tested needs to be plated on the same day, and before the compound plate is tested, it should be placed in a nitrogen cabinet at room temperature for storage.

3.细胞培养3. Cell Culture

3.1.细胞信息：采用稳定表达人源hP2X3，hP2X2/3受体的HEK-293细胞系，其hP2X3基因信息为NM_002559.4，其hP2X2/3基因信息为NM_170682.2/NM_002559.4。3.1. Cell information: HEK-293 cell line stably expressing human hP2X3 and hP2X2/3 receptors was used, and its hP2X3 gene information was NM_002559.4, and its hP2X2/3 gene information was NM_170682.2/NM_002559.4.

3.2.细胞复苏：将冻存的hP2X稳定表达HEK293细胞放置在37℃水浴中快速解冻。将冻存管外部去污后转移至生物安全柜中，吸出冻存管内细胞转移至含10mL完全培养基的离心管(成分见表9)，700g离心5分钟。去除上清后，再使用完全培养基重悬细胞。细胞均在湿润的含有5％CO2的细胞培养箱中培养，温度为37℃。3.2. Cell recovery: Place the frozen hP2X stably expressing HEK293 cells in a 37°C water bath for rapid thawing. Decontaminate the outside of the cryotube and transfer it to a biosafety cabinet. Aspirate the cells in the cryotube and transfer them to a centrifuge tube containing 10 mL of complete medium (see Table 9 for ingredients), and centrifuge at 700g for 5 minutes. After removing the supernatant, resuspend the cells in complete medium. All cells were cultured in a humidified cell culture incubator containing 5% CO 2 at 37°C.

表9

Table 9

3.3.细胞传代：正常情况下，hP2X细胞系按1∶3或1∶4的稀释比例每周传代2次。当细胞在T-75烧瓶中达到80％以及以上的融合时，使用0.25％胰蛋白酶-EDTA溶液消化细胞约2-5分钟至细胞可以自然脱落。最终根据稀释比例将细胞悬液转移到另一个T-75烧瓶中进行传代培养。3.3. Cell passaging: Under normal circumstances, hP2X cell lines are passaged twice a week at a dilution ratio of 1:3 or 1:4. When the cells reach 80% or more confluence in a T-75 flask, use 0.25% trypsin-EDTA solution to digest the cells for about 2-5 minutes until the cells can fall off naturally. Finally, transfer the cell suspension to another T-75 flask for subculture according to the dilution ratio.

4.人源P2X3，P2X2/3靶点的FLIPR检测方法4. FLIPR detection method for human P2X3 and P2X2/3 targets

4.1.细胞种板：按上述方法重悬细胞后，通过全自动细胞计数仪Cell Countess测定细胞密度和活力，计算后用完全培养基调节细胞悬液体积，将hP2X3细胞以10,000细胞/孔，将hP2X2/3细胞以18,000细胞/孔(30μL/孔)左右的密度接种于PDL包被的黑色底透384孔板上。板子会放入加湿空气控制(5％CO2)的37℃培养箱中过夜培养。4.1. Cell seeding: After resuspending the cells as described above, the cell density and viability were determined by the fully automated cell counter Cell Countess. After calculation, the volume of the cell suspension was adjusted with complete culture medium, and hP2X3 cells were seeded at a density of 10,000 cells/well and hP2X2/3 cells at a density of about 18,000 cells/well (30 μL/well) on a PDL-coated black bottom transparent 384-well plate. The plate was placed in a humidified air controlled (5% CO 2 ) 37°C incubator for overnight culture.

4.2.FLIPR读板：从培养箱中取出过夜培养的细胞板，弃掉培养基后加入Ca6染料(Molecular Devices，美国)。每孔加入25μL Ca6，孵育2h。最后将细胞板、化合物板、激动剂板和FLIPR配套384孔板枪头放入FLIPRPENTA(Molecular Devices，美国)中，设置钙流检测激发光为470-515nm，发射光为515-575nm，校正本底信号窗口至700-800RFU后，运行FLIPR程序。具体程序为读取基线60s后，FLIPR枪头从3倍浓度的化合物板中吸取12.5μL加入细胞板，90s后再从4倍浓度的刺激板中吸取12.5μL加入细胞板。加入刺激剂后，数据记录至少持续3分钟，采样率设置为1Hz。原始数据以加入激动剂后ΔF/F的最大值导出分析。4.2. FLIPR plate reading: Take out the cell plate cultured overnight from the incubator, discard the culture medium and add Ca6 dye (Molecular Devices, USA). Add 25 μL Ca6 to each well and incubate for 2 hours. Finally, put the cell plate, compound plate, agonist plate and FLIPR matching 384-well plate tip into FLIPR PENTA (Molecular Devices, USA), set the calcium flow detection excitation light to 470-515nm, the emission light to 515-575nm, correct the background signal window to 700-800RFU, and run the FLIPR program. The specific procedure is to read the baseline for 60 seconds, then the FLIPR tip draws 12.5 μL from the 3-fold concentration compound plate and adds it to the cell plate, and then draws 12.5 μL from the 4-fold concentration stimulation plate and adds it to the cell plate after 90 seconds. After adding the stimulant, the data recording lasts for at least 3 minutes, and the sampling rate is set to 1Hz. The raw data is exported and analyzed as the maximum value of ΔF/F after the addition of the agonist.

5.质量控制：以平均值和标准差进行复孔计算。窗口由最大信号值除以最小信号值得到，超过2.5倍本次测试可被接受。以Z因子作为性能指标，超过0.5可被接受。5. Quality control: The average value and standard deviation are used for duplicate calculation. The window is obtained by dividing the maximum signal value by the minimum signal value. If it exceeds 2.5 times, the test is acceptable. The Z factor is used as a performance indicator, and if it exceeds 0.5, it is acceptable.

6.数据分析：数据采用Excel 2013(Microsoft)和GraphPad Prism 7.0进行分析。用相对荧光强度倍数导出数据后，数据用下列公式进行标准化：化合物抑制率％＝(化合物光强倍数-0.5％DMSO光强倍数平均数)/(EC80激动的光强倍数-0.5％DMSO光强倍数平均数)*100。通过GraphPadPrism 5.0，采用四参数非线性回归进行量效关系的曲线拟合和IC50。如果最低测试浓度得到的药效大于50％，或最高测试浓度得到的药效小于50％，则IC50分别报告为小于最低测试浓度，或大于最高测试浓度。6. Data analysis: Data were analyzed using Excel 2013 (Microsoft) and GraphPad Prism 7.0. After exporting the data using relative fluorescence intensity multiples, the data were standardized using the following formula: Compound inhibition rate % = (compound light intensity multiple - average of 0.5% DMSO light intensity multiples) / (EC 80 excited light intensity multiple - average of 0.5% DMSO light intensity multiples) * 100. GraphPadPrism 5.0 was used to perform curve fitting of the dose-effect relationship and IC 50 using four-parameter nonlinear regression. If the lowest test concentration obtained an efficacy greater than 50%, or the highest test concentration obtained an efficacy less than 50%, then the IC 50 is reported as less than the lowest test concentration, or greater than the highest test concentration, respectively.

实验结果见表10。The experimental results are shown in Table 10.

表10化合物对P2X3，P2X2/3的实验结果
Table 10 Experimental results of compounds on P2X3, P2X2/3

结论：本发明的化合物对P2X3受体具有优异的抑制活性。Conclusion: The compounds of the present invention have excellent inhibitory activity on P2X3 receptors.

实验例2：体内药代动力学性质研究Experimental Example 2: In vivo pharmacokinetic properties study

实验目的：测定化合物在Hartley豚鼠中的药代动力学参数。Experimental purpose: To determine the pharmacokinetic parameters of the compound in Hartley guinea pigs.

实验材料：Experimental Materials:

Hartley豚鼠(雄性，400-500g，6～9周龄，北京维通利华)Hartley guinea pigs (male, 400-500 g, 6-9 weeks old, Beijing Weitong Lihua)

实验方法：Experimental methods:

1.该项目使用4只雄性Hartley豚鼠，给药前称重，根据体重，计算给药量，然后将大鼠分成两组。一 组2只Hartley豚鼠进行静脉注射给药，给药剂量为1mg/kg，给药浓度0.5mg/mL；另外一组2只Hartley豚鼠进行口服给药，给药剂量为3mg/kg，给药浓度0.5mg/mL；1. This project uses 4 male Hartley guinea pigs. They are weighed before administration and the dosage is calculated based on their body weight. The rats are then divided into two groups. Two Hartley guinea pigs in one group were intravenously injected with the drug at a dose of 1 mg/kg and a concentration of 0.5 mg/mL; another group of two Hartley guinea pigs were orally administered with a dose of 3 mg/kg and a concentration of 0.5 mg/mL;

2.收集给药后0.083(仅静脉组)、0.25、0.5、1、2、4、8、24h的血浆样品。每个样品采集约0.05mL，肝素钠抗凝，采集后放置湿冰上。2. Collect plasma samples at 0.083 (only intravenous group), 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. Each sample is about 0.05 mL, anticoagulated with sodium heparin, and placed on wet ice after collection.

3.血液样本采集后置于冰上，并于1小时之内离心分离血浆(离心条件：6000g，3分钟，2-8℃)。血浆样本在分析前存放时则放于-80℃冰箱内。3. After blood samples were collected, they were placed on ice and centrifuged within 1 hour to separate plasma (centrifugation conditions: 6000g, 3 minutes, 2-8°C). Plasma samples were stored in a -80°C refrigerator before analysis.

4.对收集的样品进行LC-MS/MS分析并采集数据。采集的分析数据用Phoenix WinNonlin 8.2.0软件计算相关药代动力学参数，如达峰浓度(Cmax)，清除率(Cl)，半衰期(T1/2)，药时曲线下面积(AUC)，生物利用度(Bioavailability)等。4. Perform LC-MS/MS analysis on the collected samples and collect data. The collected analytical data are used to calculate relevant pharmacokinetic parameters such as peak concentration (C max ), clearance (Cl), half-life (T 1/2 ), area under the drug-time curve (AUC), bioavailability, etc.

实验结果：部分实验结果见表11。Experimental results: Some experimental results are shown in Table 11.

表11体内药代动力学实验结果
Table 11 In vivo pharmacokinetic experimental results

结论：本发明化合物具有较好的暴露量和生物利用度。Conclusion: The compounds of the present invention have good exposure and bioavailability.

实验例3考察受试药物对豚鼠咳嗽模型的治疗作用Experimental Example 3: Investigating the therapeutic effect of the test drug on the guinea pig cough model

实验目的Purpose

考察目标化合物对豚鼠咳嗽的治疗作用，对ATP+柠檬酸诱导的豚鼠咳嗽的影响进行动物实验，证明受试药物可以降低咳嗽次数。The therapeutic effect of the target compound on guinea pig cough was investigated, and animal experiments were conducted on the effect on ATP+citric acid-induced guinea pig cough, which proved that the test drug could reduce the number of coughs.

实验动物Experimental animals

表12实验动物基本信息

Table 12 Basic information of experimental animals

实验分组Experimental Grouping

柠檬酸造模组、ATP+柠檬酸造模、ATP+柠檬酸造模给药1组、ATP+柠檬酸造模给药2组、ATP+柠檬酸造模给药3组，ATP+柠檬酸造模给药4组，动物数：8只/组。Citric acid modeling group, ATP+citric acid modeling, ATP+citric acid modeling and administration group 1, ATP+citric acid modeling and administration group 2, ATP+citric acid modeling and administration group 3, ATP+citric acid modeling and administration group 4, number of animals: 8/group.

实验方法Experimental methods

1.1实验流程1.1 Experimental Procedure

豚鼠造模前2小时，灌胃给与对应药物。按照分组方式分别进行柠檬酸和ATP+柠檬酸造模。柠檬酸造模组，柠檬酸雾化10分钟，统计柠檬酸10分钟雾化、停止雾化观察5、10分钟豚鼠咳嗽次数；ATP+柠檬酸造模组，使用ATP雾化2分钟，柠檬酸雾化10分钟，统计10分钟雾化、停止雾化观察5、10分钟豚鼠咳嗽次数。Guinea pigs were gavaged with the corresponding drugs 2 hours before modeling. Citric acid and ATP+citric acid modeling were performed according to the grouping method. In the citric acid modeling group, citric acid was nebulized for 10 minutes, and the number of guinea pig coughs was counted after 10 minutes of citric acid nebulization and 5 and 10 minutes of observation after stopping nebulization; in the ATP+citric acid modeling group, ATP was nebulized for 2 minutes and citric acid was nebulized for 10 minutes, and the number of guinea pig coughs was counted after 10 minutes of nebulization and 5 and 10 minutes of observation after stopping nebulization.

1.2给药1.2 Administration

1.2.1药剂配制1.2.1 Preparation of medicine

使用20％PEG400+10％Solutol+70％双蒸水进行溶解。Use 20% PEG400+10% Solutol+70% double distilled water for dissolution.

1.2.2给药方法及剂量(见表13)1.2.2 Administration method and dosage (see Table 13)

表13
Table 13

给药时间：造模前2小时，灌胃给药。Administration time: 2 hours before modeling, by intragastric administration.

1.3造模方法1.3 Modeling method

柠檬酸造模：造模前，将豚鼠放入雾化给药箱中适应10分钟。适应完毕后，调节给药装置，按照0.6mL/min的雾化速率，雾化给与0.1M柠檬酸，雾化10分钟后停止雾化。Citric acid modeling: Before modeling, the guinea pigs were placed in the atomization drug delivery box for 10 minutes to adapt. After the adaptation was completed, the drug delivery device was adjusted to atomize 0.1 M citric acid at an atomization rate of 0.6 mL/min, and the atomization was stopped after 10 minutes of atomization.

ATP+柠檬酸造模：造模前，将豚鼠放入雾化给药箱中适应10分钟，最后2分钟调节给药装置，按照0.6mL/min的雾化速率，雾化给与10μMATP，ATP雾化结束后，按照0.6mL/min的雾化速率，雾化给与0.1M柠檬酸，雾化10分钟后停止雾化。ATP+citric acid modeling: Before modeling, the guinea pigs were placed in the nebulizer drug delivery box to adapt for 10 minutes. The drug delivery device was adjusted in the last 2 minutes to administer 10 μM ATP by nebulization at a nebulization rate of 0.6 mL/min. After the ATP nebulization was completed, 0.1 M citric acid was administered by nebulization at a nebulization rate of 0.6 mL/min. The nebulization was stopped after 10 minutes of nebulization.

1.4检测指标1.4 Detection indicators

1.4.1豚鼠咳嗽统计1.4.1 Guinea pig cough statistics

柠檬酸雾化开始统计豚鼠咳嗽，统计柠檬酸雾化10分钟，停止雾化观察5、10分钟豚鼠咳嗽次数，豚鼠咳嗽声响亮，统计听到的次数，第一次咳嗽的时间。Count the number of guinea pig coughs when citric acid nebulization starts, count the number of times guinea pig coughs after 10 minutes of citric acid nebulization, stop nebulization and observe for 5 and 10 minutes. The guinea pig coughs loudly, and count the number of times heard, as well as the time of the first cough.

1.5统计分析1.5 Statistical analysis

数据采用Graphpad Prism 9(Version 9.4.0)进行分析与作图，Adobe Illustrator 2022(Version 26.3.1)进行整理合图。所有数据均以means±SD表示，组间统计学差异采用one-way ANOVA和Tukey检验，P值小 于0.05认为有显著性差异。The data were analyzed and plotted using Graphpad Prism 9 (Version 9.4.0), and the figures were organized and combined using Adobe Illustrator 2022 (Version 26.3.1). All data are expressed as means ± SD, and statistical differences between groups were analyzed using one-way ANOVA and Tukey test. The difference was considered significant at 0.05.

研究结果如表14所示：The research results are shown in Table 14:

表14
Table 14

ATP雾化2分钟，柠檬酸雾化10分钟后，观察5分钟、10分钟后的咳嗽次数；通入雾化柠檬酸后，到第一次咳嗽的时间，记为各组豚鼠咳嗽潜伏期。N＝8，结果以Mean±SD形式呈现，与CA组相比，*p＜0.05，**p＜0.01，***p＜0.001，****p＜0.0001；与ATP+CA组相比，#p＜0.05，##p＜0.01，###p＜0.001，####p＜0.0001After ATP nebulization for 2 minutes and citric acid nebulization for 10 minutes, the number of coughs after 5 minutes and 10 minutes was observed; the time from the introduction of nebulized citric acid to the first cough was recorded as the cough latency of guinea pigs in each group. N = 8, the results are presented in the form of Mean ± SD, compared with the CA group, *p < 0.05, **p < 0.01, ***p < 0.001, ****p <0.0001; compared with the ATP + CA group, # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001

实验结论：本发明化合物能显著降低ATP+柠檬酸诱导豚鼠咳嗽次数，且能显著升高ATP+柠檬酸诱导豚鼠咳嗽潜伏期。 Experimental conclusion: The compound of the present invention can significantly reduce the number of coughs induced by ATP+citric acid in guinea pigs, and can significantly increase the latency of coughs induced by ATP+citric acid in guinea pigs.

Claims (23)

式(III)所示化合物、其立体异构体或其药学上可接受的盐，其选自：
The compound represented by formula (III), its stereoisomer or a pharmaceutically acceptable salt thereof, which is selected from:
其中，in, R1选自H、D、卤素、CN、C1-3烷基或C1-3卤代烷基；R 1 is selected from H, D, halogen, CN, C 1-3 alkyl or C 1-3 haloalkyl; 各R2分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、-CH2OH、环丙基、C1-3烷基和C1-3烷氧基，所述环丙基、C1-3烷基和C1-3烷氧基任选被1、2或3个Ra取代；each R 2 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , -CH 2 OH, cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy, wherein the cyclopropyl, C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 Ra ; 各R3分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基，所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rb取代；each R 3 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R b ; 各R4分别独立地选自H、D、F、Cl、Br、I、CN、NH2、OH、CO2H、-CO2CH3、C1-3烷基和C1-3烷氧基，所述C1-3烷基和C1-3烷氧基任选被1、2或3个Rc取代；each R 4 is independently selected from H, D, F, Cl, Br, I, CN, NH 2 , OH, CO 2 H, -CO 2 CH 3 , C 1-3 alkyl and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R c ; R5和R6各自独立地选自H、D、F、CN和C1-4烷基；R 5 and R 6 are each independently selected from H, D, F, CN and C 1-4 alkyl; 或者，R5和R6及相连的碳原子形成环丙基；Alternatively, R 5 and R 6 and the carbon atom to which they are connected form a cyclopropyl group; R7和R8各自独立地选自H、D、F、OH、CN和C1-4烷基；R 7 and R 8 are each independently selected from H, D, F, OH, CN and C 1-4 alkyl; 或者，R7和R8及相连的碳原子形成环丙基、环丁基或氧杂环丁基；Alternatively, R7 and R8 and the carbon atoms to which they are connected form a cyclopropyl group, a cyclobutyl group or an oxetanyl group; R9选自OH、C1-4烷氧基、C1-4烷氨基和所述C1-4烷氧基和C1-4烷氨基各自独立地任选被1、2或3个选自F、Cl、Br、I和OH的取代基取代；R 9 is selected from OH, C 1-4 alkoxy, C 1-4 alkylamino and The C 1-4 alkoxy and C 1-4 alkylamino groups are each independently optionally substituted by 1, 2 or 3 substituents selected from F, Cl, Br, I and OH; 各Ra分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each Ra is independently selected from D, F, Cl, Br, I, CN, NH2 and OH; 各Rb分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R b is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH; 各Rc分别独立地选自D、F、Cl、Br、I、CN、NH2和OH；Each R c is independently selected from D, F, Cl, Br, I, CN, NH 2 and OH; n选自0、1、2、3和4；n is selected from 0, 1, 2, 3 and 4; m选自0、1、2、3和4；m is selected from 0, 1, 2, 3 and 4; r选自0、1、2、3和4；r is selected from 0, 1, 2, 3 and 4; t选自1、2和3；t is selected from 1, 2 and 3; 环A1选自苯基和5-10元杂芳基；Ring A1 is selected from phenyl and 5-10 membered heteroaryl; 环A2选自苯基和5-6元杂芳基；Ring A2 is selected from phenyl and 5-6 membered heteroaryl; 环A3选自苯基和5-6元杂芳基；Ring A3 is selected from phenyl and 5-6 membered heteroaryl; E1不存在，或选自NR10；E 1 is absent or selected from NR 10 ; R10选自H和C1-4烷基；R 10 is selected from H and C 1-4 alkyl; 当t为2，且E1不存在时，R7、R8不全是H。When t is 2 and E1 does not exist, R7 and R8 are not all H. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R1选自H、D、F、Cl和甲 基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from H, D, F, Cl and M base. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，各R2分别独立地选自H、D、F、Cl、CN、甲基和甲氧基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein each R 2 is independently selected from H, D, F, Cl, CN, methyl and methoxy. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，各R3分别独立地选自H、D、F、Cl和甲基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein each R 3 is independently selected from H, D, F, Cl and methyl. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，各R4分别独立地选自H、D、F、Cl和甲基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein each R 4 is independently selected from H, D, F, Cl and methyl. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R5和R6各自独立地选自H、D、F和甲基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R 5 and R 6 are each independently selected from H, D, F and methyl. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，选自-CH2-、-(CH2)2-、-CH(CH3)-、-C(CH3)2-、-(CH2)3-、-CH2CH(CH3)-、-CH2C(CH3)2-和-CH2CH(F)-；The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Selected from -CH 2 -, -(CH 2 ) 2 -, -CH(CH 3 )-, -C(CH 3 ) 2 -, -(CH 2 ) 3 -, -CH 2 CH(CH 3 )-, -CH 2 C(CH 3 ) 2 -, and -CH 2 CH(F)-; 或者，选自-CH2-、-CH(CH3)-、-CH2CH(CH3)-和-CH2C(CH3)2-。or, Selected from -CH 2 -, -CH(CH 3 )-, -CH 2 CH(CH 3 )- and -CH 2 C(CH 3 ) 2 -. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，R9选自OH、OCH3、NHS(O)2CH3、NHCH3、NHCH2CHF2和NHCH2CF3。The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, wherein R 9 is selected from OH, OCH 3 , NHS(O) 2 CH 3 , NHCH 3 , NHCH 2 CHF 2 and NHCH 2 CF 3 . 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，环A1选自苯基、 The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein ring A1 is selected from phenyl, 或者，环A1选自 Alternatively, ring A1 is selected from 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，环A2选自苯基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein ring A2 is selected from phenyl. 根据权利要求1所述化合物、其立体异构体或其药学上可接受的盐，其中，环A3选自苯基。The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein Ring A3 is selected from phenyl. 根据权利要求1-9任意一项所述化合物、其立体异构体或其药学上可接受的盐，其选自：
The compound according to any one of claims 1 to 9, its stereoisomer or a pharmaceutically acceptable salt thereof, which is selected from:
其中，T1选自CH和N；wherein T 1 is selected from CH and N; R1、R2、R3、R4、R5、R6、R7、R8、R9、m、n、r和t如权利要求1-9任意一项所定义。R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , m, n, r and t are as defined in any one of claims 1 to 9. 下列化合物、其立体异构体或其药学上可接受的盐，





The following compounds, stereoisomers thereof or pharmaceutically acceptable salts thereof,





下列化合物、其立体异构体或其药学上可接受的盐，





The following compounds, stereoisomers thereof or pharmaceutically acceptable salts thereof,





化合物TM03的B晶型，其特征在于，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°、14.23±0.20°、19.33±0.20°和20.91±0.20°：
The B crystal form of the compound TM03 is characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 19.33±0.20° and 20.91±0.20°:
根据权利要求15所述的B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°、14.23±0.20°、17.16±0.20°、19.33±0.20°、20.03±0.20°、20.91±0.20°、21.46±0.20°和23.91±0.20°。The crystal form B according to claim 15, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 17.16±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20° and 23.91±0.20°. 根据权利要求15所述的B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°、14.23±0.20°、14.60±0.20°、17.16±0.20°、19.33±0.20°、20.03±0.20°、20.91±0.20°、21.46±0.20°、23.91±0.20°、24.49±0.20°、25.15±0.20°和27.14±0.20°。The crystal form B according to claim 15, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 14.60±0.20°, 17.16±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20°, 23.91±0.20°, 24.49±0.20°, 25.15±0.20° and 27.14±0.20°. 根据权利要求15所述的B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°、14.23±0.20°、14.60±0.20°、17.16±0.20°、19.33±0.20°、20.03±0.20°、20.91±0.20°、21.46±0.20°、21.90±0.20°、22.36±0.20°、23.60±0.20°、23.91±0.20°、24.49±0.20°、25.15±0.20°、27.14±0.20°和29.89±0.20°。The crystal form B according to claim 15, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 14.23±0.20°, 14.60±0.20°, 17.16±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20°, 21.90±0.20°, 22.36±0.20°, 23.60±0.20°, 23.91±0.20°, 24.49±0.20°, 25.15±0.20°, 27.14±0.20° and 29.89±0.20°. 根据权利要求15所述的B晶型，其X射线粉末衍射图谱在下列2θ角处具有特征衍射峰：7.06±0.20°、9.40±0.20°、10.83±0.20°、12.39±0.20°、14.23±0.20°、14.60±0.20°、15.31±0.20°、15.99±0.20°、16.24±0.20°、17.16±0.20°、18.96±0.20°、19.33±0.20°、20.03±0.20°、20.91±0.20°、21.46±0.20°、21.90±0.20°、22.36±0.20°、23.60±0.20°、23.91±0.20°、24.49±0.20°、24.88±0.20°、25.15±0.20°、26.66±0.20°、27.14±0.20°、27.78±0.20°、28.29±0.20°、29.22±0.20°、29.44±0.20°、29.89±0.20°、31.48±0.20°、32.39±0.20°、33.22±0.20°和34.96±0.20°。The crystal form B according to claim 15, whose X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.06±0.20°, 9.40±0.20°, 10.83±0.20°, 12.39±0.20°, 14.23±0.20°, 14.60±0.20°, 15.31±0.20°, 15.99±0.20°, 16.24±0.20°, 17.16±0.20°, 18.96±0.20°, 19.33±0.20°, 20.03±0.20°, 20.91±0.20°, 21.46±0.20° , 21.90±0.20°, 22.36±0.20°, 23.60±0.20°, 23.91±0.20°, 24.49±0.20°, 24.88±0.20°, 25.15±0.20°, 26.66±0.20°, 27.14±0.20°, 27.78±0.20°, 28.29±0.20°, 29.22±0.20°, 29.44±0.20°, 29.89±0.20°, 31.48±0.20°, 32.39±0.20°, 33.22±0.20° and 34.96±0.20°. 化合物TM03的B晶型，其XRPD图谱基本上如图9所示。The XRPD spectrum of Form B of compound TM03 is basically as shown in FIG9 . 根据权利要求15～20任意一项所述的B晶型，其特征在于，具备下列任意一项特征：The crystal form B according to any one of claims 15 to 20, characterized in that it has any one of the following characteristics: (1)其差示扫描量热曲线在187.33℃±5℃处具有吸热峰的起始点；(1) Its differential scanning calorimetry curve has an endothermic peak starting point at 187.33°C ± 5°C; (2)其DSC图谱基本上如图10所示；(2) Its DSC spectrum is substantially as shown in FIG10 ; (3)其热重分析曲线在30-200℃时无失重；(3) Its thermogravimetric analysis curve shows no weight loss at 30-200°C; (4)其TGA图谱基本上如图11所示。(4) Its TGA spectrum is basically as shown in Figure 11. 根据权利要求1～14任意一项所述的化合物、其立体异构体或其药学上可接受的盐或根据权利要求15～21所述的化合物TM03的B晶型在制备治疗P2X3抑制剂相关疾病的药物上的应用。Use of the compound according to any one of claims 1 to 14, its stereoisomer or pharmaceutically acceptable salt thereof, or the B crystal form of the compound TM03 according to claims 15 to 21 in the preparation of a medicament for treating diseases related to P2X3 inhibitors. 根据权利要求22所述的应用，其特征在于，所述P2X3抑制剂相关疾病是指难治性咳嗽和/或COVID-19相关咳嗽。 The use according to claim 22 is characterized in that the P2X3 inhibitor-related disease refers to refractory cough and/or COVID-19-related cough.
PCT/CN2024/076273 2023-02-15 2024-02-06 Pyrimidinone derivative and pharmaceutical application thereof Ceased WO2024169781A1 (en)

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