WO2021160134A1 - 3-芳氧基-3-五元杂芳基-丙胺类化合物的制备方法及晶型 - Google Patents

3-芳氧基-3-五元杂芳基-丙胺类化合物的制备方法及晶型 Download PDF

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WO2021160134A1
WO2021160134A1 PCT/CN2021/076337 CN2021076337W WO2021160134A1 WO 2021160134 A1 WO2021160134 A1 WO 2021160134A1 CN 2021076337 W CN2021076337 W CN 2021076337W WO 2021160134 A1 WO2021160134 A1 WO 2021160134A1
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compound
formula
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unsubstituted
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French (fr)
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于娟
王友鑫
殷婷婷
曾金香
张玲玲
丁强
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Shanghai Leado Pharmatech Co Ltd
Zhangzhou Pientzehuang Pharmaceutical Co Ltd
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Shanghai Leado Pharmatech Co Ltd
Zhangzhou Pientzehuang Pharmaceutical Co Ltd
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Priority to CN202180013974.1A priority Critical patent/CN115135644B/zh
Priority to EP21754062.4A priority patent/EP4105212A4/en
Priority to JP2022549204A priority patent/JP7446591B2/ja
Priority to US17/799,487 priority patent/US12492195B2/en
Publication of WO2021160134A1 publication Critical patent/WO2021160134A1/zh
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • 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
    • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
    • C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
    • C07D407/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur 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
    • C07D413/02—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
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs

Definitions

  • the invention relates to the fields of medicinal chemistry and pharmacotherapy, in particular to a preparation method and crystal form of a 3-aryloxy-3-pentavalent heteroaryl-propylamine compound.
  • Pain is known as the fifth vital sign, and it is a warning sign of damage to body tissues. Pain is one of the most common reasons for patients to seek medical treatment. According to the duration, it is divided into acute pain (rapid onset, short duration, or continuous state) and chronic pain (slow onset or transformed from acute pain, with long duration. It can also show intermittent attacks, and many chronic pain can not find obvious damage). Acute pain is mostly nociceptive pain caused by tissue trauma, including post-operative pain, trauma, post-burn pain, childbirth pain, angina, biliary colic, renal colic and other visceral pain, fracture pain, toothache, cancer pain, etc. Surgery and post-traumatic pain are the most common clinical acute pain and the most urgently needed treatment.
  • Chronic pain mainly includes neuropathic pain, painful osteoarthritis, chronic low back pain and vascular pain.
  • Trigeminal neuralgia, diabetic pain, sciatica or post-zoster neuralgia are the main types of neuropathic pain.
  • the global prevalence of neuropathic pain is about 10%, the incidence is high, and the patient population is large. In the United States, 10%-30% of people suffer from chronic pain, resulting in an annual social expenditure of approximately US$635 billion, more than the combined amount of cancer and heart disease.
  • Chronic pain has complex causes and is a refractory disease. Only less than 50% of patients can achieve effective analgesia through drug treatment. It is estimated that the total market size of neuralgia drugs in China in 2026 will be close to 26 billion yuan, and the market size of ion channel neuropathic pain drugs will exceed 20 billion yuan.
  • Opioids have strong analgesic effects, but long-term use can easily lead to tolerance, dependence and addiction, and have adverse reactions such as respiratory depression and central sedation.
  • Non-steroidal anti-inflammatory drugs only exert a moderate analgesic effect, but also have gastrointestinal bleeding and cardiotoxicity.
  • the National Security Council recently released a report on preventable deaths, showing that for the first time in American history, deaths caused by opioid overdose exceeded the proportion of deaths caused by car accidents. According to the Commission's analysis of accidental death data in 2017, 1 in 96 Americans died from an opioid overdose, and the number of deaths from a car accident is 1 in 103 Americans. The abuse of opioids has caused a serious social crisis that is sweeping across the United States. Therefore, the market needs a new mechanism of analgesics.
  • TRPA1 is a member of the TRP ion channel superfamily and the only member of the TRPA subfamily. It is a non-selective cation channel and is permeable to Na + , K + , Ca 2+ and Mg 2+ .
  • TRPA1 is mainly distributed on the primary sensory neurons of the dorsal root nerve (DRG), trigeminal nerve (TG) and vagus nerve (VG). From the perspective of the distributed human system, TRPA1 is highly expressed in the peripheral nervous system, respiratory system, gastrointestinal system and urinary system. When these organs and tissues have abnormal functions, the expression and function of TRPA1 channels are usually abnormal at the same time.
  • TRPA1 can convert cold stimulation, chemical stimulation and mechanical stimulation into inward current, trigger a series of physiological functions, and participate in the formation of a variety of pain sensations. Inflammatory pain is a common problem of certain chronic diseases, and clinically, there is still a lack of effective treatment methods. Animal experimental studies have shown that TRPA1 is involved in inflammatory response and plays an important role in inflammatory pain. By using TRPA1 specific blockers, the inflammatory pain response in rats can be significantly reduced. From the current research, TRPA1 plays an important role in the occurrence of asthma and cough. Compounds that induce asthma and cough, whether it is cell endogenous factors or exogenous factors, can activate TRPA1. Antagonists of TRPA1 can reduce asthma symptoms and block airway hyperresponsiveness.
  • TRPA1 participates in the regulation of visceral hypersensitivity and plays an important role in visceral pain.
  • Neurogenic pain is a pain syndrome caused by damage or disease of the central or peripheral nervous system. It mainly manifests as hyperalgesia, allodynia, and spontaneous pain.
  • TRPA1 channels play an important role in different neurogenic pain, such as diabetic neuropathy and neuropathy caused by chemotherapy drugs.
  • TRPA1 also has a mediating role in toothache, migraine and other pains. The administration of TRPA1 antagonists can significantly alleviate the pain symptoms.
  • TRPA1 is widely distributed and expressed in the human system. In addition to the physiological functions of TRPA1, the development of TRPA1 inhibitor indications reported so far also involves inflammatory bowel disease, chronic obstructive pulmonary disease, antitussive, antipruritic, Allergic rhinitis, ear disease, anti-diabetic, urinary incontinence, etc. TRPA1 is a proven new target for the treatment of many diseases.
  • the purpose of the present invention is to provide a compound with a novel structure and a TRP channel as a target (especially a TRPA1 target), and a preparation method, crystal form and use thereof.
  • Ring A is a substituted or unsubstituted 4-12 membered carbocyclic ring, a substituted or unsubstituted 4-12 membered heterocyclic ring, a substituted or unsubstituted 5-12 membered heteroaromatic ring, a substituted or unsubstituted C 6 -C 12 Aromatic ring
  • R 1 and R 2 are each independently hydrogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted 5-12 membered heteroaryl, A substituted or unsubstituted C 6 -C 12 aryl group;
  • X and Y are each independently a carbon atom, an oxygen atom, a sulfur atom or a nitrogen atom;
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted Substituted C 6 -C 12 aryl;
  • R 4 is halogen
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkane Group, substituted or unsubstituted 5-12 membered heteroaryl group, substituted or unsubstituted C 6 -C 12 aryl group;
  • W is O or S
  • n 1, 2 or 3;
  • heterocyclic ring, heteroaromatic ring and heteroaryl group each independently have 1 to 3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S.
  • Ring A is a substituted or unsubstituted 5-10 membered carbocyclic ring, a substituted or unsubstituted 5-10 membered heterocyclic ring, a substituted or unsubstituted 5-12 membered heteroaromatic ring, a substituted or unsubstituted 5-12 membered heteroaromatic ring, Substituted C 6 -C 12 aromatic ring.
  • ring A is a substituted or unsubstituted 5-10 membered carbocyclic ring, a substituted or unsubstituted 5-10 membered heterocyclic ring, and a substituted or unsubstituted 5-12 membered heteroaromatic ring.
  • ring A is not a benzene ring.
  • ring A is a substituted or unsubstituted 5-7 membered carbocyclic ring, a substituted or unsubstituted 5-7 membered heterocyclic ring, and a substituted or unsubstituted 5-7 membered heteroaromatic ring.
  • ring A is a substituted or unsubstituted 5-7 membered carbocyclic ring or a 5-7 membered heteroaromatic ring.
  • ring A is a substituted or unsubstituted 5-membered carbocyclic ring, a substituted or unsubstituted 6-membered carbocyclic ring, or a substituted or unsubstituted furan ring.
  • ring A is a 5-membered carbocyclic ring, a 6-membered carbocyclic ring or a furan ring.
  • ring A is
  • connection structure between ring A and the adjacent benzene ring is:
  • At least one of X and Y is a heteroatom.
  • Y is a carbon atom or a nitrogen atom.
  • X is S or O.
  • X is S.
  • Y is a carbon atom.
  • R 1 and R 2 are each independently hydrogen, substituted or unsubstituted C 1 -C 8 alkyl, substituted or unsubstituted C 3 -C 8 cycloalkyl, substituted or unsubstituted 5 -10 membered heteroaryl, substituted or unsubstituted C 6 -C 12 aryl.
  • R 1 and R 2 are each independently hydrogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 7 cycloalkyl.
  • R 1 and R 2 are each independently hydrogen or substituted or unsubstituted C 1 -C 3 alkyl.
  • R 1 and R 2 are each independently hydrogen, methyl or ethyl.
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 10 cycloalkyl, substituted or unsubstituted 5-12 member Heteroaryl, substituted or unsubstituted C 6 -C 12 aryl.
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 7 cycloalkyl.
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 6 alkyl.
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 4 alkyl.
  • R 3 is a hydrogen atom, a chlorine atom or a methyl group.
  • the halogen is F, Cl, Br or I.
  • R 4 is F, Cl, Br or I.
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen, substituted or unsubstituted C 1 -C 10 alkyl, substituted or unsubstituted C 3 -C 8 cycloalkyl, substituted or unsubstituted 5-10 membered heteroaryl, substituted or unsubstituted C 6 -C 12 aryl.
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 7 cycloalkyl.
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen.
  • W is O or S.
  • W is O.
  • n 1 or 2.
  • n 1
  • each R 3 is the same or different.
  • any one of the "substitution” refers to a substituent in which 1-4 (preferably 1, 2, 3, or 4) hydrogen atoms on the group are each independently selected from the following group Substituted: C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 1 -C 3 haloalkyl, halogen, nitro, cyano, hydroxyl, C 1 -C 4 carboxyl, C 2 -C 4 Ester group, C 2 -C 4 amide group, C 1 -C 6 alkoxy, C 1 -C 6 haloalkoxy, benzyl, six-membered aryl, five- or six-membered heteroaryl (preferably C 5 heteroaryl).
  • the The structure is:
  • the compound of formula a has a structure of formula a-1:
  • the compound of formula b has a structure of formula b-1:
  • the compound of formula I has the structure of formula I-A:
  • the compound of formula I is selected from the following group:
  • the compound of formula I is selected from the following group:
  • the first solvent is selected from the group consisting of dimethyl sulfoxide, toluene, DMF, or a combination thereof.
  • the first solvent is selected from the group consisting of dimethyl sulfoxide.
  • the first catalyst is selected from the group consisting of halide salts, tetrabutylammonium bromide, 4-dimethylaminopyridine, dibenzo18 crown ether 6, pyridine, or a combination thereof.
  • the halide salt is selected from the group consisting of potassium fluoride, potassium chloride, potassium bromide, potassium iodide, sodium fluoride, sodium chloride, sodium bromide, sodium iodide, or a combination thereof .
  • the first catalyst includes potassium iodide.
  • the first alkaline reagent is selected from the group consisting of inorganic bases, organic bases, or a combination thereof.
  • the first alkaline reagent is selected from the following group: sodium hydrogen, hydroxide, or a combination thereof.
  • the hydroxide is selected from the group consisting of sodium hydroxide, potassium hydroxide, or a combination thereof.
  • the first alkaline reagent includes hydroxide.
  • the first alkaline reagent includes sodium hydroxide.
  • the reaction temperature is 30-90°C, preferably 30-80°C, more preferably 40-70°C, and most preferably 50-60°C .
  • the reaction time is 10-48h, preferably 16-40h, more preferably 20-35h, most preferably 22-25h.
  • the molar ratio of the compound of formula a to the compound of formula b is 1:1-3, preferably 1:1-2, more preferably 1:1.2-1.7.
  • the molar ratio of the compound of formula a to the first alkaline reagent is 1:2.5-10, preferably 1:3-8, more preferably 1:4-6.
  • the molar ratio of the compound of formula b to the first alkaline reagent is 1-3:2.5-10, preferably 1-2:3-8, more preferably 1.2-1.7:4-6.
  • the molar ratio of the first catalyst to the first alkaline reagent is 1:5-70, preferably 1:10-60, more preferably 1:15-50, more Preferably 1:20-50, more preferably 1:25-40, and best 1:28-36.
  • the molar ratio of the first catalyst to the compound of formula a is 1:1-50, preferably 1:2-40, more preferably 1:3-30, more preferably Ground 1:5-25, better 1:5-20, best 1:5-15.
  • the reaction in the step (1), is carried out under normal pressure (such as 1 atm).
  • the reaction is carried out under N 2 protection.
  • the step (1) includes the steps:
  • the compound of formula a is dissolved in the first solvent, the compound of formula b and the first catalyst are added, the first alkali reagent is added after the temperature is lowered, and the reaction is heated to obtain the compound of formula I.
  • the temperature after cooling is 5-30°C (preferably 10-20°C).
  • the temperature after raising the temperature is 30-90°C, preferably 30-80°C, more preferably 40-70°C, and most preferably 50-60°C.
  • step (1) after the reaction is completed, water and ethyl acetate are added to the reaction solution for extraction, the organic phase is collected, and the compound of formula I is obtained by separation.
  • the compound of formula I can be isolated.
  • the acid aqueous solution is an oxalic acid aqueous solution.
  • Ring A is a substituted or unsubstituted 4-12 membered carbocyclic ring, a substituted or unsubstituted 4-12 membered heterocyclic ring, a substituted or unsubstituted 5-12 membered heteroaromatic ring, a substituted or unsubstituted C 6 -C 12 Aromatic ring
  • R 1 and R 2 are each independently a substituted or unsubstituted C 1 -C 6 alkyl group, a substituted or unsubstituted C 3 -C 7 cycloalkyl group;
  • X and Y are each independently a carbon atom, an oxygen atom, a sulfur atom or a nitrogen atom;
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted Substituted C 6 -C 12 aryl;
  • R 4 is halogen
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkane Group, substituted or unsubstituted 5-12 membered heteroaryl group, substituted or unsubstituted C 6 -C 12 aryl group;
  • R 12 is substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted C 6 -C 16 aryl, substituted or unsubstituted 5- 16-membered heteroaryl, substituted or unsubstituted C 1 -C 6 alkyl-W-, substituted or unsubstituted C 3 -C 12 cycloalkyl-W-, substituted or unsubstituted C 6 -C 16 aryl Group -W-, substituted or unsubstituted 5-16 membered heteroaryl -W-;
  • R 13 is halogen
  • W is O or S
  • n 1, 2 or 3;
  • heterocyclic ring, heteroaromatic ring and heteroaryl group each independently have 1 to 3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S.
  • ring A, X, Y, W, n, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 Each is independently as described in the first aspect of the present invention.
  • the first catalyst and the first alkaline reagent in the first solvent are independently as described in the first aspect of the present invention.
  • the compound of formula a, the compound of formula b and the compound of formula I are each independently as described in the first aspect of the present invention.
  • step (1) is as described in the first aspect of the present invention.
  • the second solvent is selected from the group consisting of dimethyl sulfoxide, toluene, or DMF, or a combination thereof.
  • the second solvent is toluene.
  • the second alkaline reagent is selected from the group consisting of triethanolamine (TEA), 1,8-diazabicycloundec-7-ene (DBU), N,N-di Isopropylethylamine (DIEA), sodium carbonate, or a combination thereof.
  • TAA triethanolamine
  • DBU 1,8-diazabicycloundec-7-ene
  • DIEA N,N-di Isopropylethylamine
  • sodium carbonate or a combination thereof.
  • the second alkaline reagent includes N,N-diisopropylethylamine (DIEA).
  • DIEA N,N-diisopropylethylamine
  • the reaction temperature is 20-80°C, preferably 30-70°C, more preferably 30-60°C, more preferably 40-50°C , Best 40 ⁇ 45°C.
  • the reaction time is 2-16h, preferably 2-10h, more preferably 2-6h, most preferably 3-5h.
  • the N-demethylating reagent represented by formula c includes phenyl chloroformate.
  • the molar ratio of the compound of formula I to the second alkaline reagent is 1:0.2-10, preferably 1:0.5-8, more preferably 1:0.8-5, more preferably 1:1-3, best 1:1-2.
  • the molar ratio of the compound of formula I to the compound of formula c is 1:0.2-10, preferably 1:0.5-8, and more Preferably, it is 1:0.8-5, more preferably 1:1-3, more preferably 1:1.1 ⁇ 2.5, and most preferably 1:1-2.
  • the molar ratio of the second alkaline reagent to the compound of formula c is 0.8-5: 0.8-5, preferably 1-3: 1-3, more preferably 1-2: 1.1-2.5, most preferably 1-2: 1.1-2.
  • the reaction in the step (2), is carried out under normal pressure (such as 1 atm).
  • the reaction is carried out under N 2 protection.
  • the step (2) includes the steps:
  • the compound of formula I and the second alkaline reagent are dissolved in the second solution, the N-demethylation reagent shown in formula c is added after cooling, and the reaction is raised to obtain the compound of formula I-a.
  • the temperature after cooling is 10-40°C, preferably 20-30°C.
  • the temperature after raising the temperature is 20-80°C, preferably 30-70°C, more preferably 30-60°C, more preferably 40-50°C, and most preferably 40-45°C .
  • step (2) after the reaction is completed, water and ethyl acetate are added to the reaction solution for extraction, the organic phase is collected, and concentrated to obtain the compound of formula I-a.
  • the compound of formula I-a has the structure of formula I-a-1:
  • the third solvent is selected from the group consisting of dimethyl sulfoxide, toluene, or DMF, or a combination thereof.
  • the third solvent is dimethyl sulfoxide.
  • the third alkaline reagent is selected from the group consisting of inorganic bases, organic bases, or a combination thereof.
  • the third alkali reagent is selected from the group consisting of potassium tert-butoxide, potassium carbonate, hydroxide, or a combination thereof.
  • the hydroxide is selected from the group consisting of sodium hydroxide, potassium hydroxide, or a combination thereof.
  • the third alkaline reagent includes hydroxide.
  • the third alkaline reagent includes sodium hydroxide.
  • the reaction temperature is 20-100°C, preferably 30-90°C, more preferably 40-80°C, more preferably 50-70°C , Best 60 ⁇ 65°C.
  • the reaction time is 5-16h, preferably 5-10h, more preferably 7-10h.
  • the molar ratio of the compound of formula Ia to the third base reagent is 1:1-20, preferably 1:2-15, more preferably 1:1 : 2-10, more preferably 1:3.5-8, best 1:6.
  • step (3) after the reaction is completed, water and ethyl acetate are added to the reaction solution for extraction, the organic phase is collected, and the compound of formula I is obtained by separation.
  • the compound of formula I can be isolated.
  • the acid aqueous solution is an oxalic acid aqueous solution.
  • the compound of formula I-1 has the structure of formula I-1-1:
  • the method includes the steps:
  • Ring A is a substituted or unsubstituted 4-12 membered carbocyclic ring, a substituted or unsubstituted 4-12 membered heterocyclic ring, a substituted or unsubstituted 5-12 membered heteroaromatic ring, a substituted or unsubstituted C 6 -C 12 Aromatic ring
  • R 1 and R 2 are each independently a substituted or unsubstituted C 1 -C 6 alkyl group, a substituted or unsubstituted C 3 -C 7 cycloalkyl group;
  • X and Y are each independently a carbon atom, an oxygen atom, a sulfur atom or a nitrogen atom;
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted Substituted C 6 -C 12 aryl;
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkane Group, substituted or unsubstituted 5-12 membered heteroaryl group, substituted or unsubstituted C 6 -C 12 aryl group;
  • R 12 is substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted C 6 -C 16 aryl, substituted or unsubstituted 5- 16-membered heteroaryl, substituted or unsubstituted C 1 -C 6 alkyl-W-, substituted or unsubstituted C 3 -C 12 cycloalkyl-W-, substituted or unsubstituted C 6 -C 16 aryl Group -W-, substituted or unsubstituted 5-16 membered heteroaryl -W-;
  • R 13 is halogen
  • W is O or S
  • n 1, 2 or 3;
  • heterocyclic ring, heteroaromatic ring and heteroaryl group each independently have 1 to 3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S.
  • rings A, X, Y, W, n, R 1 , R 2 , R 3 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently As described in the first aspect of the present invention.
  • the second catalyst and the N-demethylation reagent represented by formula c are each independently as described in the second aspect of the present invention.
  • step (2) is as described in the second aspect of the present invention.
  • the compound of formula I, the compound of formula c and the compound of formula I-a are each independently as described in the second aspect of the present invention.
  • the fourth aspect of the present invention provides a salt of the compound of formula I,
  • Ring A is a substituted or unsubstituted 5-7 membered carbocyclic ring, a substituted or unsubstituted 5-7 membered heterocyclic ring, a substituted or unsubstituted 5-7 membered heteroaromatic ring;
  • R 1 and R 2 are each independently hydrogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 7 cycloalkyl;
  • X and Y are each independently a carbon atom, an oxygen atom, a sulfur atom or a nitrogen atom;
  • W is O or S
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 7 cycloalkyl;
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 3 -C 7 cycloalkane base;
  • n 1, 2 or 3;
  • heterocyclic ring, heteroaromatic ring and heteroaryl group each independently have 1 to 3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S.
  • rings A, X, Y, W, n, R 1 , R 2 , R 3 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently As described in the first aspect of the present invention.
  • the compound of formula I has the structure shown in formula I-B:
  • the compound of formula I has the structure shown in formula I-D:
  • the compound of formula I is selected from the following group:
  • the salt of the compound of formula I is a salt formed by the compound of formula I and an acid selected from the group consisting of hydrochloric acid, mucic acid, D-glucuronic acid, hydrobromic acid, hydrofluoric acid, and sulfuric acid. , Nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, toluene methanesulfonic acid , Benzenesulfonic acid, aspartic acid, glutamic acid, or a combination thereof.
  • the salt of the compound of formula I is selected from the group consisting of hydrochloride, maleate, oxalate, mucate, fumarate, D-glucuronate, or Its combination.
  • the molar ratio of the compound of formula Ic-1 to hydrochloric acid is 4:1, 3:1, 2:1, 1:1 , 1:2, 1:3 or 4:1.
  • the hydrochloride crystal form A of the compound of formula Ic-1 is an anhydrous crystal form.
  • the X-ray powder diffraction pattern of the hydrochloride crystal form A has characteristic peaks at 2 ⁇ angles of 18.27 ⁇ 0.2°, 21.27 ⁇ 0.2°, and 22.89 ⁇ 0.2°.
  • the hydrochloride crystal form A also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 16.85 ⁇ 0.2°, 22.20 ⁇ 0.2°, 23.86 ⁇ 0.2°.
  • the hydrochloride crystal form A also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 10.09 ⁇ 0.2°, 25.40 ⁇ 0.2°, 28.18 ⁇ 0.2°.
  • the hydrochloride crystal form A also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 10.09 ⁇ 0.2°, 16.85 ⁇ 0.2°, 22.20 ⁇ 0.2°, 23.86 ⁇ 0.2°, 25.40 ⁇ 0.2°, 28.18 ⁇ 0.2°.
  • the hydrochloride crystal form A also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 11.25 ⁇ 0.2°, 21.84 ⁇ 0.2°, 26.76 ⁇ 0.2°, 28.75 ⁇ 0.2°, 32.57 ⁇ 0.2°.
  • the hydrochloride crystal form A has characteristic peaks at one or more 2 ⁇ values selected from the following group: 10.09 ⁇ 0.2°, 11.25 ⁇ 0.2°, 16.85 ⁇ 0.2°, 18.27 ⁇ 0.2°, 21.27 ⁇ 0.2°, 21.84 ⁇ 0.2°, 22.20 ⁇ 0.2°, 22.89 ⁇ 0.2°, 23.86 ⁇ 0.2°, 25.40 ⁇ 0.2°, 26.76 ⁇ 0.2°, 28.18 ⁇ 0.2°, 28.75 ⁇ 0.2°, 32.57 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the hydrochloride crystal form A has one or a characteristic peak and peak intensity at 2 ⁇ values selected from the following group:
  • the hydrochloride salt crystal form A has characteristic peaks of X-ray powder diffraction substantially as shown in FIG. 8.
  • the differential scanning calorimetry (DSC) chart of the hydrochloride crystal form A is heated to 141.8 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the hydrochloride crystal form A is basically as shown in FIG. 9.
  • thermogravimetric analysis (TGA) chart of the hydrochloride crystal form A has about 1.1 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) chart of the hydrochloride crystal form A is basically as shown in FIG. 9.
  • the first organic solvent includes ethyl acetate.
  • the hydrochloric acid is concentrated hydrochloric acid.
  • the pH of the system is 6.5-7.5, preferably 7.0.
  • the reaction time is 3-8 min, preferably 5 min.
  • the reaction in the step (a), is carried out under stirring conditions.
  • step (a) the hydrochloric acid is slowly added.
  • the weight-to-volume ratio (kg:L) of the compound of formula Ic-1 to the first organic solvent is 0.05-2:1-20, preferably 0.05-1 :1-10, more preferably 0.1-0.5: 2-7.
  • step (a) after the solid precipitates, it is dried at 40-45° C. to obtain the hydrochloride crystal form A of the compound of formula Ic-1.
  • the molar ratio of the compound of formula Ic-1 to maleic acid is 4:1, 3:1, 2:1, 1:1, 1:2, 1:3 or 4:1.
  • the maleate crystal form B of the Ic-1 compound is an anhydrous crystal form.
  • the X-ray powder diffraction pattern of the maleate salt crystal form B has characteristic peaks at 2 ⁇ angles of 19.23 ⁇ 0.2°, 24.04 ⁇ 0.2°, and 24.70 ⁇ 0.2°.
  • the maleate salt crystal form B also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 11.83 ⁇ 0.2°, 19.56 ⁇ 0.2°, 28.15 ⁇ 0.2°.
  • the maleate salt crystal form B also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 9.70 ⁇ 0.2°, 18.23 ⁇ 0.2°, 24.93 ⁇ 0.2°.
  • the maleate salt crystal form B also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 9.70 ⁇ 0.2°, 11.83 ⁇ 0.2°, 18.23 ⁇ 0.2°, 19.56 ⁇ 0.2°, 24.93 ⁇ 0.2°, 28.15 ⁇ 0.2°.
  • the maleate salt crystal form B also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 15.22 ⁇ 0.2°, 16.30 ⁇ 0.2°, 18.82 ⁇ 0.2°, 21.15 ⁇ 0.2°, 21.83 ⁇ 0.2°, 23.60 ⁇ 0.2°, 26.35 ⁇ 0.2°, 28.94 ⁇ 0.2°, 32.59 ⁇ 0.2°, 33.31 ⁇ 0.2°, 34.74 ⁇ 0.2°, 35.94 ⁇ 0.2°, 38.18.
  • the maleate salt crystal form B has characteristic peaks at one or more 2 ⁇ values selected from the following group: 9.70 ⁇ 0.2°, 11.83 ⁇ 0.2°, 15.22 ⁇ 0.2°, 16.30 ⁇ 0.2°, 18.23 ⁇ 0.2°, 18.82 ⁇ 0.2°, 19.23 ⁇ 0.2°, 19.56 ⁇ 0.2°, 21.15 ⁇ 0.2°, 21.83 ⁇ 0.2°, 23.60 ⁇ 0.2°, 24.04 ⁇ 0.2°, 24.70 ⁇ 0.2°, 24.93 ⁇ 0.2°, 26.35 ⁇ 0.2°, 28.15 ⁇ 0.2°, 28.94 ⁇ 0.2°, 32.59 ⁇ 0.2°, 33.31 ⁇ 0.2°, 34.74 ⁇ 0.2°, 35.94 ⁇ 0.2°, 38.18 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the maleate salt crystal form B has one or a characteristic peak and peak intensity at 2 ⁇ values selected from the following group:
  • the maleate salt crystal form B has characteristic peaks of X-ray powder diffraction substantially as shown in FIG. 11.
  • the differential scanning calorimetry (DSC) chart of the maleate crystal form B is heated to 105.8 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the maleate salt crystal form B is basically as shown in FIG. 12.
  • thermogravimetric analysis (TGA) chart of the maleate crystal form B has a value of about 0.8 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2%) when heated to 80°C. Or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) diagram of the maleate salt crystal form B is basically as shown in FIG. 12.
  • the second organic solvent includes ethyl acetate.
  • the reaction time is 4-10 days, preferably 5-7 days.
  • the reaction is carried out under stirring conditions.
  • the rotation speed of the stirring is 700-800 rpm.
  • the reaction temperature is room temperature.
  • the weight-to-volume ratio (kg:ml) of the compound of formula Ic-1 to the second organic solvent is 0.05-2:1-10, preferably 0.05-1 :1-30, more preferably 0.1-0.5:1-20.
  • the weight of the compound of formula Ic-1 and maleic acid is 0.05-2:0.05-0.3, preferably 0.05-1:0.05-0.2, more preferably 0.1-0.5: 0.08-0.16.
  • step (b) after the solid precipitates, it is dried under vacuum at room temperature to obtain the maleate salt crystal form B of the compound of formula Ic-1.
  • the ninth aspect of the present invention provides an oxalate or oxalate crystal form C of the compound of formula Ic-1,
  • the molar ratio of the compound of formula Ic-1 to oxalic acid is 4:1, 3:1, 2:1, 1:1 , 1:2, 1:3 or 4:1.
  • the oxalate crystal form C of the Ic-1 compound is an anhydrous crystal form.
  • the X-ray powder diffraction pattern of the oxalate crystal form C has characteristic peaks at 2 ⁇ angles of 14.64 ⁇ 0.2°, 22.05 ⁇ 0.2°, and 25.61 ⁇ 0.2°.
  • the oxalate crystal form C also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 16.36 ⁇ 0.2°, 20.90 ⁇ 0.2°, 23.43 ⁇ 0.2°.
  • the oxalate crystal form C also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 15.27 ⁇ 0.2°, 16.07 ⁇ 0.2°, 19.52 ⁇ 0.2°.
  • the oxalate crystal form C also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 15.27 ⁇ 0.2°, 16.07 ⁇ 0.2°, 16.36 ⁇ 0.2°, 17.63 ⁇ 0.2°, 19.52 ⁇ 0.2°, 20.90 ⁇ 0.2°, 23.43 ⁇ 0.2°.
  • the oxalate crystal form C has characteristic peaks at one or more 2 ⁇ values selected from the following group: 14.64 ⁇ 0.2°, 15.27 ⁇ 0.2°, 16.07 ⁇ 0.2°, 16.36 ⁇ 0.2°, 17.63 ⁇ 0.2°, 19.52 ⁇ 0.2°, 20.90 ⁇ 0.2°, 22.05 ⁇ 0.2°, 23.43 ⁇ 0.2°, 25.61 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the oxalate crystal form C has one or a characteristic peak and peak intensity at 2 ⁇ values selected from the following group:
  • the oxalate crystal form C has characteristic X-ray powder diffraction peaks as shown in FIG. 15.
  • the differential scanning calorimetry (DSC) chart of the oxalate crystal form C is heated to 152.2 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the oxalate crystal form C is basically as shown in FIG. 16.
  • thermogravimetric analysis (TGA) chart of the oxalate crystal form C has about 1.0 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) diagram of the oxalate crystal form C is basically as shown in FIG. 16.
  • the tenth aspect of the present invention provides a method for preparing the oxalate crystal form C of the compound of formula Ic-1 described in the ninth aspect of the present invention, and the method comprises the steps:
  • the third organic solvent includes ethyl acetate.
  • the reaction time is 4-10 days, preferably 5-7 days.
  • the reaction in the step (c), is carried out under stirring conditions.
  • the rotation speed of the stirring is 700-800 rpm.
  • the reaction temperature is room temperature.
  • the weight-to-volume ratio (kg:ml) of the compound of formula Ic-1 to the third organic solvent is 0.05-2:1-10, preferably 0.05-1 :1-30, more preferably 0.1-0.5:1-20.
  • the weight of the compound of formula Ic-1 and oxalic acid is 0.05-2:0.05-0.3, preferably 0.08-1:0.05-0.2, more preferably 0.1- 0.5: 0.06-0.13.
  • step (c) after the solid is precipitated, it is vacuum dried at room temperature to obtain the oxalate crystal form C of the compound of formula Ic-1.
  • the eleventh aspect of the present invention provides a mucate or mucate crystal form D of the compound of formula Ic-1,
  • the molar ratio of the compound of the formula Ic-1 to the mucic acid is 4:1, 3:1, 2:1, 1: 1, 1:2, 1:3 or 4:1.
  • the mucic acid salt crystal form D of the Ic-1 compound is an anhydrous crystal form.
  • the X-ray powder diffraction pattern of the Mucic acid salt crystal form D has characteristic peaks at 2 ⁇ angles of 3.79 ⁇ 0.2°, 11.28 ⁇ 0.2°, and 19.48 ⁇ 0.2°.
  • the mucate salt crystal form D also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 15.81 ⁇ 0.2°, 20.98 ⁇ 0.2°, 23.91 ⁇ 0.2°.
  • the mucate salt crystal form D also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 16.97 ⁇ 0.2°, 25.88 ⁇ 0.2°, 28.40 ⁇ 0.2°.
  • the mucate salt crystal form D also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 15.81 ⁇ 0.2°, 16.97 ⁇ 0.2°, 20.98 ⁇ 0.2°, 23.91 ⁇ 0.2°, 25.88 ⁇ 0.2°, 28.40 ⁇ 0.2°.
  • the mucate salt crystal form D also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 14.21 ⁇ 0.2°, 17.71 ⁇ 0.2°, 27.16 ⁇ 0.2°, 29.49 ⁇ 0.2°, 30.74 ⁇ 0.2°, 32.33 ⁇ 0.2°, 34.50 ⁇ 0.2°, 35.42 ⁇ 0.2°, 36.16 ⁇ 0.2°.
  • the mucic acid salt crystal form D has characteristic peaks at one or more 2 ⁇ values selected from the following group: 3.79 ⁇ 0.2°, 11.28 ⁇ 0.2°, 14.21 ⁇ 0.2°, 15.81 ⁇ 0.2°, 16.97 ⁇ 0.2°, 17.71 ⁇ 0.2°, 19.48 ⁇ 0.2°, 20.98 ⁇ 0.2°, 23.91 ⁇ 0.2°, 25.88 ⁇ 0.2°, 27.16 ⁇ 0.2°, 28.40 ⁇ 0.2°, 29.49 ⁇ 0.2°, 30.74 ⁇ 0.2°, 32.33 ⁇ 0.2°, 34.50 ⁇ 0.2°, 35.42 ⁇ 0.2°, 36.16 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the crystalline form D of mucic acid salt has one or a characteristic peak and peak intensity at 2 ⁇ values selected from the following group:
  • the crystalline form D of the mucate salt has characteristic X-ray powder diffraction peaks as shown in FIG. 18.
  • the differential scanning calorimetry (DSC) chart of the crystalline form D of mucic acid is heated to 140.9 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the crystalline form D of the mucic acid salt is basically as shown in FIG. 19.
  • thermogravimetric analysis (TGA) chart of the crystalline form D of mucic acid has about 1.45 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) chart of the crystalline form D of the mucic acid salt is basically as shown in FIG. 19.
  • the twelfth aspect of the present invention provides a method for preparing the crystalline form D of the compound of formula Ic-1 described in the eleventh aspect of the present invention, the method comprising the steps:
  • the fourth organic solvent includes ethyl acetate.
  • the reaction temperature is 40-60°C.
  • the reaction time is 1-3 days, preferably 1.5-2.5 days.
  • the weight-to-volume ratio (mg:ml) of mucic acid to the compound of formula Ic-1 is 1-20:0.1-1.5, preferably 1-15:0.1 -1, more preferably 5-10: 0.2-0.8.
  • the concentration of the compound of formula Ic-1 in the fourth solvent is 20-60 mg/mL, preferably 30-50 mg/mL , More preferably 35-45mg/mL.
  • step (d) after the solid precipitates, it is vacuum dried at room temperature to obtain the mucic acid crystal form D of the compound of formula Ic-1.
  • the thirteenth aspect of the present invention provides a fumarate or fumarate crystal form E of the compound of formula Ic-1,
  • the molar ratio of the compound of formula Ic-1 to fumaric acid is 4:1, 3:1, 2:1, 1:1, 1:2, 1:3 or 4:1.
  • the fumarate crystal form E of the Ic-1 compound is an anhydrous crystal form.
  • the X-ray powder diffraction pattern of the fumarate crystal form E has characteristic peaks at 2 ⁇ angles of 14.17 ⁇ 0.2°, 18.95 ⁇ 0.2°, and 23.76 ⁇ 0.2°.
  • the fumarate crystal form E also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 4.7 ⁇ 0.2°, 22.75 ⁇ 0.2°, 26.93 ⁇ 0.2°.
  • the fumarate crystal form E also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 13.34 ⁇ 0.2°, 15.63 ⁇ 0.2°, 28.69 ⁇ 0.2°.
  • the fumarate crystal form E also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 4.7 ⁇ 0.2°, 13.34 ⁇ 0.2°, 15.63 ⁇ 0.2°, 22.75 ⁇ 0.2°, 26.93 ⁇ 0.2°, 28.69 ⁇ 0.2°.
  • the fumarate crystal form E also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 9.41 ⁇ 0.2°, 17.62 ⁇ 0.2°, 25.66 ⁇ 0.2°, 31.34 ⁇ 0.2°.
  • the fumarate salt crystal form E has characteristic peaks at one or more 2 ⁇ values selected from the following group: 4.7 ⁇ 0.2°, 9.41 ⁇ 0.2°, 13.34 ⁇ 0.2°, 14.17 ⁇ 0.2°, 15.63 ⁇ 0.2°, 17.62, 18.95 ⁇ 0.2°, 22.75 ⁇ 0.2°, 23.76 ⁇ 0.2°, 25.66 ⁇ 0.2°, 26.93 ⁇ 0.2°, 28.69 ⁇ 0.2°, 31.34 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the fumarate crystal form E has one or a characteristic peak and peak intensity at 2 ⁇ values selected from the following group:
  • the fumarate salt crystal form E has characteristic X-ray powder diffraction peaks substantially as shown in FIG. 21.
  • the differential scanning calorimetry (DSC) chart of the fumarate crystal form E is heated to 76.5 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the fumarate crystal form E is basically as shown in FIG. 22.
  • thermogravimetric analysis (TGA) chart of the fumarate crystal form E has about 2.17 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2%) when heated to 80°C. Or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) chart of the fumarate crystal form E is basically as shown in FIG. 22.
  • the fourteenth aspect of the present invention provides a method for preparing the fumarate crystal form E of the compound of formula Ic-1 described in the thirteenth aspect of the present invention, the method comprising the steps:
  • the fifth organic solvent includes ethyl acetate.
  • the mixture is stirred at room temperature for 3-5 days.
  • the reaction temperature is room temperature
  • the weight-to-volume ratio (mg:ml) of fumaric acid to the compound of formula Ic-1 is 1-20:0.1-1.5, preferably 1-15: 0.1-1, more preferably 5-10: 0.2-0.8.
  • the concentration of the compound of formula Ic-1 in the fifth solvent is 20-60 mg/mL, preferably 30-50 mg/mL , More preferably 35-45mg/mL.
  • step (e) after the solid precipitates, it is vacuum dried at room temperature to obtain the fumaric acid crystal form E of the compound of formula Ic-1.
  • the fifteenth aspect of the present invention provides a D-glucuronate or D-glucuronate crystal form F of the compound of formula Ic-1,
  • the molecular molar ratio of the compound of formula Ic-1 to D-glucuronic acid is 4:1, 3:1, 2:1, 1 :1, 1:2, 1:3 or 4:1.
  • the D-glucuronate crystal form F of the Ic-1 compound is an anhydrous crystal form.
  • the X-ray powder diffraction pattern of the D-glucuronate crystal form F has characteristic peaks at 2 ⁇ angles of 4.77 ⁇ 0.2°, 16.13 ⁇ 0.2°, and 19.53 ⁇ 0.2°.
  • the D-glucuronate crystal form F also has characteristic peaks of 8.34 ⁇ 0.2°, 17.54 ⁇ 0.2°, 20.06 ⁇ 0.2° at one or more 2 ⁇ values selected from the following group .
  • the D-glucuronate crystal form F also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 10.87 ⁇ 0.2°, 21.25 ⁇ 0.2°, 25.93 ⁇ 0.2 °.
  • the D-glucuronate crystal form F also has characteristic peaks at one or more 2 ⁇ values selected from the following group: 8.34 ⁇ 0.2°, 10.87 ⁇ 0.2°, 17.54 ⁇ 0.2 °, 20.06 ⁇ 0.2°, 21.25 ⁇ 0.2°, 23.42 ⁇ 0.2°, 25.93 ⁇ 0.2°.
  • the D-glucuronate crystal form F has characteristic peaks at one or more 2 ⁇ values selected from the following group: 4.77 ⁇ 0.2°, 8.34 ⁇ 0.2°, 10.87 ⁇ 0.2° , 16.13 ⁇ 0.2°, 17.54 ⁇ 0.2°, 19.53 ⁇ 0.2°, 20.06 ⁇ 0.2°, 21.25 ⁇ 0.2°, 23.42 ⁇ 0.2°, 25.93 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the D-glucuronate crystal form F has one or a characteristic peak and peak intensity at 2 ⁇ values selected from the following group:
  • the D-glucuronate crystal form F has characteristic X-ray powder diffraction peaks as shown in FIG. 24.
  • the differential scanning calorimetry (DSC) chart of the D-glucuronate crystal form F is heated to 119.1 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C). °C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the D-glucuronate crystal form F is basically as shown in FIG. 25.
  • thermogravimetric analysis (TGA) chart of the D-glucuronate crystal form F has about 2.71 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) diagram of the D-glucuronate crystal form F is basically as shown in FIG. 25.
  • the sixteenth aspect of the present invention is a method for preparing the D-glucuronate crystal form F of the compound of formula Ic-1 according to the fifteenth aspect of the present invention, said method comprising the steps:
  • the sixth organic solvent includes acetonitrile.
  • the mixture is stirred at room temperature for 3-5 days.
  • the reaction temperature is room temperature
  • the weight-to-volume ratio (mg:ml) of D-glucuronic acid to the compound of formula Ic-1 is 5-30:0.1-2, preferably 5- 20:0.1-1, more preferably 10-16:0.2-0.7.
  • the concentration of the compound of formula Ic-1 in the fifth solvent is 20-60 mg/mL, preferably 30-50 mg/mL , More preferably 35-45mg/mL.
  • step (f) after the solid is precipitated, it is dried under vacuum at room temperature to obtain the D-glucuronic acid crystal form F of the compound of formula Ic-1.
  • the seventeenth aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the hydrochloride crystal form A of the compound of formula Ic-1 as described in the fifth aspect of the present invention, and the hydrochloride salt of the seventh aspect of the present invention
  • Salt crystal form F and a pharmaceutically acceptable carrier.
  • the eighteenth aspect of the present invention provides a hydrochloride crystal form A of the compound of formula Ic-1 according to the fifth aspect of the present invention, and the maleate of the compound of formula Ic-1 according to the seventh aspect of the present invention Form B, the oxalate crystal form C of the compound of formula Ic-1 in the ninth aspect of the present invention, the mucic acid salt form D of the compound of formula Ic-1 in the eleventh aspect of the present invention, and the first aspect of the present invention
  • TRP transient receptor potential channel protein
  • the transient receptor potential channel protein is TRPA1.
  • the disease associated with transient receptor potential channel protein is selected from the group consisting of pain, epilepsy, inflammation, respiratory disorder, pruritus, urinary tract disorder, inflammatory bowel disease, or a combination thereof.
  • the pain is acute pain or chronic pain.
  • the pain is selected from the group consisting of acute inflammatory pain, chronic inflammatory pain, visceral pain, neurogenic pain, fibromyalgia, headache, neuralgia, pain caused by cancer, or a combination thereof.
  • the pain is inflammatory pain.
  • the inflammatory pain is acute inflammatory pain or chronic inflammatory pain.
  • the headache is migraine or muscle tension pain.
  • the neuralgia is trigeminal neuralgia, diabetic pain or post-zoster neuralgia.
  • the pain is selected from the following group: acute pain, fibromyalgia, visceral pain, inflammatory pain, neuralgia, or a combination thereof.
  • the pain is fibromyalgia.
  • the nineteenth aspect of the present invention provides a non-therapeutic and non-diagnostic in vitro method for inhibiting the activity of transient receptor potential channel proteins, which comprises the steps of: combining transient receptor potential channel proteins or cells expressing the proteins with such
  • the hydrochloride crystal form A of the compound of formula Ic-1 according to the fifth aspect of the invention, the maleate crystal form B of the compound of formula Ic-1 according to the seventh aspect of the invention, and the crystalline form B of the compound of formula Ic-1 according to the ninth aspect of the invention The oxalate crystal form C of the compound of formula Ic-1, the mucic acid salt form D of the compound of formula Ic-1 described in the eleventh aspect of the present invention, the compound of formula Ic-1 described in the thirteenth aspect of the present invention
  • the fumarate crystal form E or the D-glucuronate crystal form F of the compound of formula Ic-1 described in the fifteenth aspect of the present invention is contacted, thereby inhibiting the activity of transient receptor potential channel protein.
  • the twentieth aspect of the present invention provides a method for inhibiting transient receptor potential channel protein or preventing and/or treating diseases related to transient receptor potential channel protein (TRP), comprising the steps of: administering such as the present invention to a subject in need
  • the hydrochloride crystal form A of the compound of formula Ic-1 according to the fifth aspect of the invention, the maleate crystal form B of the compound of formula Ic-1 according to the seventh aspect of the invention, and the crystalline form B of the compound of formula Ic-1 according to the ninth aspect of the invention The oxalate crystal form C of the compound of formula Ic-1, the mucic acid salt form D of the compound of formula Ic-1 described in the eleventh aspect of the present invention, the compound of formula Ic-1 described in the thirteenth aspect of the present invention Fumarate crystal form E, or D-glucuronate crystal form F of the compound of formula Ic-1 in the fifteenth aspect of the present invention, thereby preventing and/or treating and transient receptor potential channel protein (TRP) Related diseases.
  • TRP trans
  • the subject includes humans and non-human mammals (rodents, rabbits, monkeys, domestic animals, dogs, cats, etc.).
  • Figure 3 is the analgesic activity induced pain model in mice results in the hot plate Compound I C -23 disclosure.
  • FIG 5 Compound I C -1, duloxetine, indomethacin and the analgesic activity of choline mountain buttercup pain model in mice results torsion acetate.
  • Figure 6 shows the results of analgesic activity of compound IC-1 and gabapentin in a rat SNL model.
  • Figure 7 shows the statistical results of the addition time of compound Ic-1 and duloxetine in the phase II (10-60 min) phase at different dosages in the mouse formalin model.
  • Figure 8 is an XRPD pattern of the hydrochloride salt form A of the compound of formula Ic-1.
  • Figure 9 is a TGA/DSC chart of the hydrochloride salt form A of the compound of formula Ic-1.
  • Fig. 10 is a comparison diagram of temperature-changing XRPD of the hydrochloride salt crystal form A of the compound of formula Ic-1.
  • Figure 11 is an XRPD pattern of maleate salt form B of the compound of formula Ic-1.
  • Figure 12 is a TGA/DSC chart of the maleate salt form B of the compound of formula Ic-1.
  • Figure 13 is a 1H NMR spectrum of the maleate salt crystal form B of the compound of formula Ic-1.
  • Fig. 14 is a comparison chart of the temperature-variable XRPD of the maleate salt crystal form B of the compound of formula Ic-1.
  • Figure 15 is an XRPD pattern of the oxalate crystal form C of the compound of formula Ic-1.
  • Figure 16 is a TGA/DSC chart of oxalate form C of the compound of formula Ic-1.
  • Figure 17 is a temperature-variable XRPD comparison diagram of the oxalate crystal form C of the compound of formula Ic-1.
  • Figure 18 is the XPRD pattern of the crystalline form D of the mucic acid salt of the compound of formula Ic-1.
  • Fig. 19 is a TGA/DSC chart of the crystalline form D of the mucic acid salt of the compound of formula Ic-1.
  • Fig. 20 is a 1H NMR spectrum of the crystalline form D of the mucic acid salt of the compound of formula Ic-1.
  • Fig. 21 is an XRPD pattern of the fumarate salt form E of the compound of formula Ic-1.
  • Figure 22 is a TGA/DSC chart of the fumarate salt form E of the compound of formula Ic-1.
  • Figure 23 is a 1H NMR spectrum of the fumarate salt form E of the compound of formula Ic-1.
  • Figure 24 is an XRPD pattern of D-glucuronate salt form F of the compound of formula Ic-1.
  • Figure 25 is a TGA/DSC chart of D-glucuronate salt form F of the compound of formula Ic-1.
  • Figure 26 is a DVS diagram of the hydrochloride salt form A of the compound of formula Ic-1.
  • Figure 27 is a comparison diagram of XRPD before and after the DVS test of the hydrochloride salt form A of the compound of formula Ic-1.
  • Figure 28 is a DVS diagram of the maleate salt form B of the compound of formula Ic-1.
  • Fig. 29 is a comparison diagram of XRPD before and after DVS test of maleate salt form B of the compound of formula Ic-1.
  • Figure 30 is a comparison diagram of XRPD before and after the stability test of the hydrochloride salt form A of the compound of formula Ic-1.
  • Figure 31 is a comparison diagram of XRPD before and after the stability test of maleate salt form B of the compound of formula Ic-1.
  • Figure 32 is a single crystal diffraction pattern of the hydrochloride salt form A of the compound of formula Ic-1.
  • the present inventors unexpectedly developed a compound, or a pharmaceutically acceptable salt thereof, and a preparation method and salt crystal form thereof for the first time.
  • the compound has the structure of Formula I.
  • Experiments show that the compound of the present invention has a significant inhibitory effect on TRP channels.
  • the compounds of the present invention can effectively treat pain related to TRP (especially TRPA1) targets. On this basis, the present invention has been completed.
  • the terms “comprising”, “including”, and “containing” are used interchangeably and include not only closed definitions, but also semi-closed and open definitions. In other words, the term includes “consisting of” and “consisting essentially of”.
  • R 1 the same “R 1” and “R1" meaning, are interchangeable, the same as other similarly defined meanings.
  • C 1 -C 12 alkyl refers to a straight or branched chain alkyl group having 1-12, 1-8, 1-6, 1-4 or 1-3 carbon atoms, such as methyl, ethyl, propyl, isopropyl Group, butyl, isobutyl, sec-butyl, tert-butyl, or similar groups.
  • C 1 -C 6 alkoxy refers to a linear or branched alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy , Butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentoxy, hexyloxy, or similar groups.
  • C 6 -C 12 benzoalicyclic group refers to a group having 6-12 carbon atoms, including indanyl, tetrahydronaphthyl, dihydronaphthyl and similar groups.
  • C 3 -C 12 cycloalkyl or “C 3 -C 7 cycloalkyl” refers to a cycloalkyl group having 3-12 or 3-7 carbon atoms (including monocyclic, bicyclic Or polycyclic ring system), such as cyclopropyl, cyclobutyl, methylcyclobutyl, cyclopentyl, cycloheptyl, or similar groups.
  • C 2 -C 6 ester group refers to a group having a C 1 -C 5 alkyl-COO- structure or a group having a -COO-C 1 to C 5 alkyl structure, wherein the alkyl group It can be linear or branched, such as CH 3 COO-, C 2 H 5 COO-, C 3 H 8 COO-, (CH 3 )2CHCOO-, -COOCH 3 , -COOC 2 H 5 , -COOC 3 H 8 , or similar groups.
  • C 2 -C 4 amido group refers to a group having a C 1 -C 3 alkyl-CO-NH- structure or a group having a -CO-NH-C 1 -C 3 alkyl structure , Where the alkyl group can be straight or branched, such as CH 3 -CO-NH-, C 2 H 5 -CO-NH-, C 3 H 8 -CO-NH-, -COOCH 3 , -CO-NH -C 2 H 5 , -CO-NH-C 3 H 8 , or similar groups.
  • C 2 -C 4 acyl refers to a group having a C 1 -C 3 alkyl-CO- structure where the alkyl group can be linear or branched, such as CH 3 -CO-, C 2 H 5 -CO-, C 3 H 8 -CO-, or similar groups.
  • C 3 -C 7 heterocycloalkyl refers to having 3-7 ring carbon atoms and 1-3 heteroatoms (preferably containing 1 nitrogen atom, that is, it is commonly adjacent to R 1 and R 2 The nitrogen atom) of monocyclic and polycyclic heterocycles (preferably monocyclic heterocycles), such as piperidine group, tetrahydropyrrolyl group, or similar groups.
  • the terms "4-12 membered carbocyclic ring”, “5-10 membered carbocyclic ring” or “5-7 membered carbocyclic ring” are any stable 4, 5, 6, 7, 8, 9, 10, 11 Or a 12-membered monocyclic, bicyclic or polycyclic ring.
  • the carbocyclic ring can be a saturated, partially unsaturated, or unsaturated ring, but cannot be an aromatic ring.
  • carbocyclic ring examples include, but are not limited to, cyclopropyl ring, cyclobutene ring, cyclobutene ring, cyclopentene ring, cyclopentene ring, cyclohexene ring, cyclohexene ring, cycloheptene ring, cycloheptene ring, Adamantane ring, cyclooctane ring, cyclooctene ring, cyclooctadiene ring, bicyclo[3.3.0]octane, bicyclo[4.3.0]nonane, bicyclo[4.4.0]decane, two Cyclo[2.2.2]octane, fluorenyl, indan ring.
  • heterocyclic ring refers to any stable monocyclic, bicyclic or polycyclic ring (for example, 5, 6 or 7 membered), and the heterocyclic ring contains one or more (such as 1-3) selected from N, O
  • the heterocyclic ring can be a saturated, partially unsaturated, or unsaturated ring, but cannot be an aromatic ring.
  • the number of members in front of the heterocyclic ring refers to the number of ring atoms, for example, a 4-12 membered heterocyclic ring has 4-12 ring atoms.
  • heteromatic ring refers to an aromatic heterocyclic ring system having one to more (preferably 1, 2, or 3) heteroatoms selected from N, O, and S.
  • the number of members in front of the heteroaromatic ring refers to the number of ring atoms, for example, a 5-7 membered heteroaromatic ring has 5-7 ring atoms. It should be understood that when multiple heteroatoms are contained, the heteroatoms may be the same, part of the same, or completely different.
  • examples of 5-membered heteroaromatic rings include (but are not limited to): pyrrole ring, furan ring, thiophene ring, imidazole ring, oxazole ring, and thiazole ring.
  • examples of 6-membered heteroaromatic ring include (but are not limited to) pyridine ring, Pyrazine ring, pyridazine ring, pyrimidine ring. Or similar groups.
  • C 1 -C 6 haloalkyl and “C 1 -C 3 haloalkyl” refer to one or more of linear or branched alkyl groups having 1-6 and 1-3 carbon atoms.
  • One hydrogen atom is replaced by a halogen group, such as monochloromethane, dichloroethane, trichloropropane, or similar groups.
  • C 1 -C 6 haloalkoxy means that one or more hydrogen atoms of a linear or branched alkoxy group having 1 to 6 carbon atoms are replaced by a halogen group, such as a chloroform Oxy, dichlorooxy, or similar groups.
  • C 1 -C 4 ⁇ carboxy refers to a group of C 1 -C 3 alkyl-COOH structure, where the alkyl group can be linear or branched, such as CH 3 COOH, C 2 H 5 COOH, C 3 H 8 COOH, (CH 3 ) 2 CHCOOH, or similar groups.
  • C 6 -C 12 aryl refers to a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 12 carbon atoms in the ring portion, such as phenyl, naphthyl, biphenyl, or similar Group.
  • aromatic ring refers to an aromatic ring system.
  • the number of carbons in front of the aromatic ring refers to the number of ring atoms, for example, "C 6 -C 12 aromatic ring” refers to a monocyclic or bicyclic aromatic ring having 6 to 12 carbon atoms in the ring portion, typically, C 6-
  • the C 12 aromatic ring is a benzene ring or a naphthalene ring.
  • heteroaryl refers to an optionally substituted aromatic group, for example, it is a 5- to 7-membered monocyclic ring system having a ring containing at least one heteroatom and at least one carbon atom.
  • the number of members in front of a heteroaryl group refers to the number of ring atoms.
  • a 5-12 membered heteroaryl group refers to a heteroaryl group with 5-12 ring atoms, including (but not limited to): pyrrolyl, thienyl, pyridyl, Pyridazinyl, pyrimidinyl, pyrazinyl, furyl, imidazolyl, thiazolyl, oxazolyl, triazolyl, or similar groups.
  • halogen refers to F, Cl, Br, and I.
  • substitution refers to the substitution of a hydrogen atom on a group by a non-hydrogen atom group, but the valence requirements must be met and a chemically stable compound is generated by the substitution. In this specification, it should be construed that all substituents are unsubstituted, unless expressly described as “substituted” herein.
  • substituents can be connected to the parent group or substrate on any atom, unless the connection violates the valence requirement; the hydrogen atoms of the parent group or substrate can be on the same atom, It can also be on different atoms.
  • room temperature refers to 25 ⁇ 5°C.
  • the range includes not only the endpoints P1 and P2, but also any numerical points between the endpoints P1 and P2.
  • the value range includes any integer value points between the endpoints P1 and P2.
  • the value range is 1-10, it includes 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; the value range 3-7 includes 3, 4, and 5 , 6, 7.
  • C3-C7 includes C3, C4, C5C6, and C7.
  • Ring A is a substituted or unsubstituted 4-12 membered carbocyclic ring, a substituted or unsubstituted 4-12 membered heterocyclic ring, a substituted or unsubstituted 5-12 membered heteroaromatic ring, a substituted or unsubstituted C 6 -C 12 Aromatic ring
  • R 1 and R 2 are each independently hydrogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted 5-12 membered heteroaryl, A substituted or unsubstituted C 6 -C 12 aryl group;
  • X and Y are each independently a carbon atom, an oxygen atom, a sulfur atom or a nitrogen atom;
  • R 3 is hydrogen, halogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkyl, substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted Substituted C 6 -C 12 aryl;
  • R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently hydrogen, substituted or unsubstituted C 1 -C 12 alkyl, substituted or unsubstituted C 3 -C 12 cycloalkane Group, substituted or unsubstituted 5-12 membered heteroaryl group, substituted or unsubstituted C 6 -C 12 aryl group;
  • W is O or S
  • n 1, 2 or 3;
  • heterocyclic ring, heteroaromatic ring and heteroaryl group each independently have 1 to 3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S.
  • rings A, X, Y, W, n, R 1 , R 2 , R 3 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 and R 11 are each independently As described above in the first aspect of the present invention.
  • the compound of the present invention not only has an inhibitory effect on TRPA1, but also has a certain inhibitory effect on other members of the TRP family.
  • pharmaceutically acceptable salt refers to a salt formed by the compound of the present invention and an acid or a base suitable for use as a medicine.
  • Pharmaceutically acceptable salts include inorganic salts and organic salts.
  • a preferred type of salt is the salt formed by the compound of the present invention and an acid.
  • Acids suitable for salt formation include (but are not limited to): hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid and other inorganic acids, formic acid, acetic acid, etc.
  • a preferred type of salt is a metal salt formed by the compound of the present invention and a base.
  • Suitable bases for salt formation include (but are not limited to): sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, sodium phosphate and other inorganic bases, Organic bases such as ammonia, triethylamine, diethylamine, etc.
  • the compound of formula I described in the present invention can be converted into its pharmaceutically acceptable salt by conventional methods.
  • a solution of the corresponding acid can be added to the solution of the above compound, and the solvent can be removed under reduced pressure after the salt is formed. That is, the corresponding salt of the compound of the present invention is obtained.
  • the present invention provides a method for preparing a compound of formula I, or a pharmaceutically acceptable salt thereof.
  • the method includes the steps:
  • each reaction parameter (such as the first solvent, the first catalyst and the first alkali reagent) in the step (1) is as described in the first aspect of the present invention.
  • the present invention provides a method for preparing a compound of formula Ic-1, the method comprising the steps:
  • each reaction parameter (such as the first solvent, the first catalyst and the first alkali reagent) in the step (1) is as described in the second aspect of the present invention.
  • the present invention provides a salt crystal form of the compound of formula Ic-1.
  • hydrochloride salt form A of the compound of formula Ic-1 As used herein, the terms "hydrochloride salt form A" of the compound of formula Ic-1", “hydrochloride salt form A” and “crystal form A” are used interchangeably.
  • maleate salt form B of the compound of formula Ic-1 As used herein, the terms "maleate salt form B of the compound of formula Ic-1", “maleate salt form B” and “crystal form B” are used interchangeably.
  • oxalate crystal form C of the compound of formula Ic-1 As used herein, the terms "oxalate crystal form C of the compound of formula Ic-1", “oxalate crystal form C" and “crystal form C" are used interchangeably.
  • crystal form D of mucic acid salt of the compound of formula Ic-1 As used herein, the terms "crystal form D of mucic acid salt of the compound of formula Ic-1", “crystal form D of mucic acid salt” and “crystal form D” are used interchangeably.
  • fluarate salt form E of the compound of formula Ic-1 As used herein, the terms "fumarate salt form E of the compound of formula Ic-1", “fumarate salt form E” and “crystal form E” are used interchangeably.
  • D-glucuronate crystalline form F of the compound of formula Ic-1 As used herein, the terms "D-glucuronate crystalline form F of the compound of formula Ic-1", “D-glucuronate crystalline form F” and “crystalline form F” are used interchangeably.
  • the present invention provides a hydrochloride crystal form A of the compound of formula Ic-1, the hydrochloride crystal form A having a characteristic peak at one or more 2 ⁇ values selected from the following group: 10.09 ⁇ 0.2°, 11.25 ⁇ 0.2°, 16.85 ⁇ 0.2°, 18.27 ⁇ 0.2°, 21.27 ⁇ 0.2°, 21.84 ⁇ 0.2°, 22.20 ⁇ 0.2°, 22.89 ⁇ 0.2°, 23.86 ⁇ 0.2°, 25.40 ⁇ 0.2°, 26.76 ⁇ 0.2°, 28.18 ⁇ 0.2°, 28.75 ⁇ 0.2°, 32.57 ⁇ 0.2°.
  • the hydrochloride salt crystal form A has characteristic peaks of X-ray powder diffraction substantially as shown in FIG. 8.
  • the differential scanning calorimetry (DSC) chart of the hydrochloride crystal form A is heated to 141.8 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the hydrochloride crystal form A is basically as shown in FIG. 9.
  • thermogravimetric analysis (TGA) chart of the hydrochloride crystal form A has about 1.1 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) chart of the hydrochloride crystal form A is basically as shown in FIG. 9.
  • the present invention provides a maleate salt crystal form B of the compound of formula Ic-1, said maleate salt crystal form B having a characteristic peak at one or more 2 ⁇ values selected from the following group: 9.70 ⁇ 0.2°, 11.83 ⁇ 0.2°, 15.22 ⁇ 0.2°, 16.30 ⁇ 0.2°, 18.23 ⁇ 0.2°, 18.82 ⁇ 0.2°, 19.23 ⁇ 0.2°, 19.56 ⁇ 0.2°, 21.15 ⁇ 0.2°, 21.83 ⁇ 0.2°, 23.60 ⁇ 0.2°, 24.04 ⁇ 0.2°, 24.70 ⁇ 0.2°, 24.93 ⁇ 0.2°, 26.35 ⁇ 0.2°, 28.15 ⁇ 0.2°, 28.94 ⁇ 0.2°, 32.59 ⁇ 0.2°, 33.31 ⁇ 0.2°, 34.74 ⁇ 0.2°, 35.94 ⁇ 0.2°, 38.18 ⁇ 0.2°.
  • maleic acid is as follows:
  • the maleate salt crystal form B has characteristic peaks of X-ray powder diffraction substantially as shown in FIG. 11.
  • the differential scanning calorimetry (DSC) chart of the maleate crystal form B is heated to 105.8 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the maleate salt crystal form B is basically as shown in FIG. 12.
  • thermogravimetric analysis (TGA) chart of the maleate crystal form B has a value of about 0.8 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2%) when heated to 80°C. Or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) diagram of the maleate salt crystal form B is basically as shown in FIG. 12.
  • the present invention provides an oxalate crystal form C of the compound of formula Ic-1, the oxalate crystal form C having a characteristic peak at one or more 2 ⁇ values selected from the group: 14.64 ⁇ 0.2°, 15.27 ⁇ 0.2°, 16.07 ⁇ 0.2°, 16.36 ⁇ 0.2°, 17.63 ⁇ 0.2°, 19.52 ⁇ 0.2°, 20.90 ⁇ 0.2°, 22.05 ⁇ 0.2°, 23.43 ⁇ 0.2°, 25.61 ⁇ 0.2°.
  • Oxalic acid is also called oxalic acid, and its structure is as follows:
  • the oxalate crystal form C has characteristic X-ray powder diffraction peaks as shown in FIG. 15.
  • the differential scanning calorimetry (DSC) chart of the oxalate crystal form C is heated to 152.2 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the oxalate crystal form C is basically as shown in FIG. 16.
  • thermogravimetric analysis (TGA) chart of the oxalate crystal form C has about 1.0 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) diagram of the oxalate crystal form C is basically as shown in FIG. 16.
  • the present invention provides a crystalline form D of the compound of formula Ic-1.
  • a crystalline form D of the compound of formula Ic-1 There are characteristic peaks at: 3.79 ⁇ 0.2°, 11.28 ⁇ 0.2°, 14.21 ⁇ 0.2°, 15.81 ⁇ 0.2°, 16.97 ⁇ 0.2°, 17.71 ⁇ 0.2°, 19.48 ⁇ 0.2°, 20.98 ⁇ 0.2°, 23.91 ⁇ 0.2°, 25.88 ⁇ 0.2°, 27.16 ⁇ 0.2°, 28.40 ⁇ 0.2°, 29.49 ⁇ 0.2°, 30.74 ⁇ 0.2°, 32.33 ⁇ 0.2°, 34.50 ⁇ 0.2°, 35.42 ⁇ 0.2°, 36.16 ⁇ 0.2°.
  • the crystalline form D of the mucate salt has characteristic X-ray powder diffraction peaks as shown in FIG. 18.
  • the differential scanning calorimetry (DSC) chart of the crystalline form D of mucic acid is heated to 140.9 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the crystalline form D of the mucic acid salt is basically as shown in FIG. 19.
  • thermogravimetric analysis (TGA) chart of the crystalline form D of mucic acid has about 1.45 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) chart of the crystalline form D of the mucic acid salt is basically as shown in FIG. 19.
  • the present invention provides a fumarate crystalline form E of the compound of formula Ic-1, the fumarate crystalline form E has a characteristic peak at one or more 2 ⁇ values selected from the following group: 4.7 ⁇ 0.2°, 9.41 ⁇ 0.2°, 13.34 ⁇ 0.2°, 14.17 ⁇ 0.2°, 15.63 ⁇ 0.2°, 17.62, 18.95 ⁇ 0.2°, 22.75 ⁇ 0.2°, 23.76 ⁇ 0.2°, 25.66 ⁇ 0.2°, 26.93 ⁇ 0.2 °, 28.69 ⁇ 0.2°, 31.34 ⁇ 0.2°.
  • the fumarate salt crystal form E has characteristic X-ray powder diffraction peaks substantially as shown in FIG. 21.
  • the differential scanning calorimetry (DSC) chart of the fumarate crystal form E is heated to 76.5 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the fumarate crystal form E is basically as shown in FIG. 22.
  • thermogravimetric analysis (TGA) chart of the fumarate crystal form E has about 2.17 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2%) when heated to 80°C. Or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) chart of the fumarate crystal form E is basically as shown in FIG. 22.
  • the present invention provides a D-glucuronate crystalline form F of a compound of formula Ic-1, wherein the D-glucuronate crystalline form F is at one or more 2 ⁇ values selected from the group consisting of With characteristic peaks: 4.77 ⁇ 0.2°, 8.34 ⁇ 0.2°, 10.87 ⁇ 0.2°, 16.13 ⁇ 0.2°, 17.54 ⁇ 0.2°, 19.53 ⁇ 0.2°, 20.06 ⁇ 0.2°, 21.25 ⁇ 0.2°, 23.42 ⁇ 0.2°, 25.93 ⁇ 0.2°.
  • the D-glucuronate crystal form F has characteristic X-ray powder diffraction peaks as shown in FIG. 24.
  • the differential scanning calorimetry (DSC) chart of the D-glucuronate crystal form F is heated to 119.1 ⁇ 5°C (preferably ⁇ 4°C, ⁇ 3°C, ⁇ 2°C). °C or ⁇ 1°C) endothermic peaks begin to appear.
  • the differential scanning calorimetry (DSC) chart of the D-glucuronate crystal form F is basically as shown in FIG. 25.
  • thermogravimetric analysis (TGA) chart of the D-glucuronate crystal form F has about 2.71 ⁇ 0.5% (preferably ⁇ 0.4%, ⁇ 0.3%, 0.2% or 0.1%) weight loss.
  • thermogravimetric analysis (TGA) diagram of the D-glucuronate crystal form F is basically as shown in FIG. 25.
  • TRP Transient Receptor Potential Channel Protein
  • Transient receptor potential channel proteins are a protein superfamily composed of important cation channels existing on the cell membrane.
  • Transient receptor potential channel proteins include multiple subfamilies, such as TRPA, TRPC, TRPM, TRPV, TRPML, and TRPP subfamilies.
  • TRPA1 channel protein is related to pain, epilepsy, inflammation, respiratory disorders, itching, urinary tract disorders, inflammatory bowel disease and other diseases. TRPA1 is used to treat pain, epilepsy, inflammation, respiratory disorders, itching, urinary tract disorders, and inflammatory diseases. Targets for diseases such as bowel disease.
  • the transient receptor potential channel protein is TRPA1.
  • diseases related to transient receptor potential channel protein include (but are not limited to): pain, epilepsy, inflammation, respiratory disorders, itching, urinary tract disorders, inflammatory bowel disease, or a combination thereof .
  • TRP transient receptor potential channel protein
  • the pain is acute pain or chronic pain.
  • the pain includes (but is not limited to): acute inflammatory pain, chronic inflammatory pain, visceral pain, neurogenic pain, fibromyalgia, headache, neuralgia, pain caused by cancer, or combination.
  • the pain is inflammatory pain.
  • the inflammatory pain is acute inflammatory pain or chronic inflammatory pain.
  • the headache is migraine or muscle tension pain.
  • the neuralgia is trigeminal neuralgia, diabetic pain or post-zoster neuralgia.
  • the pain includes (but is not limited to): acute pain, fibromyalgia, visceral pain, inflammatory pain, neuralgia, or a combination thereof.
  • the pain is fibromyalgia.
  • the present invention also provides a method for inhibiting transient receptor potential channel protein (TPR) and a method for treating diseases related to transient receptor potential channel protein.
  • TPR transient receptor potential channel protein
  • the above-mentioned compound of formula I of the present invention, or a pharmaceutically acceptable salt thereof, and the hydrochloride crystal form A, maleate crystal form B, oxalate crystal form C, and mucate crystal form D of the compound of formula I , Fumarate crystal form E and D-glucuronate crystal form F can be used to inhibit transient receptor potential channel protein, thereby preventing or treating diseases related to transient receptor potential channel protein.
  • examples of diseases related to transient receptor potential channel proteins include (but are not limited to): pain, epilepsy, inflammation, respiratory disorders, pruritus, urinary tract disorders, and inflammatory bowel disease.
  • the pain includes (but is not limited to): acute inflammatory pain, chronic inflammatory pain, visceral pain, neurogenic pain, fibromyalgia, headache (such as migraine, muscle tension pain, etc.), nerves Pain (for example, trigeminal neuralgia, diabetic pain, post-zoster neuralgia, etc.), or cancer causes pain.
  • the present invention provides a non-therapeutic in vitro method for inhibiting the activity of transient receptor potential channel proteins, including, for example, in vitro culture system, the transient receptor potential channel protein or expressing the protein Cells and the compound of formula I according to the present invention, or a pharmaceutically acceptable salt thereof, or the hydrochloride crystal form A, maleate crystal form B, oxalate crystal form C, or mucate of the compound of formula I
  • the crystal form D, fumarate crystal form E or D-glucuronate crystal form F are contacted, thereby inhibiting the activity of the transient receptor potential channel protein.
  • the present invention also provides a method for inhibiting transient receptor potential channel proteins, which can be therapeutic or non-therapeutic.
  • the method includes the steps of: administering the compound of formula I of the present invention, or a pharmaceutically acceptable salt thereof, or the hydrochloride salt form A, maleate salt form B, Oxalate crystal form C, mucate crystal form D, fumarate crystal form E or D-glucuronate crystal form F.
  • the subject includes humans and non-human mammals (rodents, rabbits, monkeys, domestic animals, dogs, cats, etc.).
  • rodents rodents, rabbits, monkeys, domestic animals, dogs, cats, etc.
  • the present invention provides a composition for inhibiting the activity of transient receptor potential channel proteins.
  • composition of the present invention includes (but is not limited to): pharmaceutical composition, food composition, dietary supplement, beverage composition and the like.
  • the composition is a pharmaceutical composition
  • the pharmaceutical composition includes the compound of formula I according to the present invention, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.
  • the composition is a pharmaceutical composition
  • the pharmaceutical composition includes the hydrochloride salt crystal form A and the maleate salt crystal form of the compound of formula I according to the present invention.
  • B Oxalate crystal form C, mucate crystal form D, fumarate crystal form E or D-glucuronate crystal form F; and a pharmaceutically acceptable carrier.
  • the dosage form of the pharmaceutical composition includes (but is not limited to) oral preparations, injections, and external preparations.
  • Representatives include (but are not limited to): tablets, injections, infusions, ointments, gels, solutions, microspheres, and films.
  • pharmaceutically acceptable carrier refers to: one or more compatible solid, semi-solid, liquid or gel fillers, which are suitable for human or animal use, and must have sufficient purity and low enough toxicity.
  • Cosmetic means that the components in the pharmaceutical composition and the active ingredients of the drug and their blending with each other without significantly reducing the efficacy of the drug.
  • the carrier is not particularly limited, and materials commonly used in the art can be selected, or prepared by conventional methods, or purchased from the market.
  • pharmaceutically acceptable carriers include cellulose and its derivatives (such as methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, etc.), gelatin, talc, and solid lubricants.
  • stearic acid such as soybean oil, sesame oil, peanut oil, olive oil, etc.
  • polyols such as propylene glycol, glycerin, mannitol, sorbitol, etc.
  • emulsifiers such as Tween
  • wetting agents such as sodium lauryl sulfate
  • buffering agents such as chelating agents, thickening agents, pH adjusters, penetration enhancers, coloring agents, flavoring agents, stabilizers, antioxidants, preservatives , Bacteriostatic agent, pyrogen-free water, etc.
  • the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, and acetic acid. Ethyl ester, propylene glycol, 1,3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.
  • the composition may also contain adjuvants such as wetting agents, emulsifying agents and suspending agents, etc.
  • the pharmaceutical preparation should match the mode of administration.
  • the agents of the present invention can also be used together with other co-therapeutics (including before, during or after use).
  • a safe and effective amount of the drug is administered to the desired subject (such as a human or non-human mammal).
  • the safe and effective amount is usually at least about 10 ⁇ g/kg body weight, and in most cases No more than about 8 mg/kg body weight, preferably the dosage is about 10 micrograms/kg body weight to about 1 mg/kg body weight.
  • the specific dosage should also consider factors such as the route of administration and the patient's health status, which are all within the skill range of a skilled physician.
  • the present invention provides a class of compounds of formula I with novel structure and excellent TRP channel inhibitory activity.
  • the compound of the present invention has excellent in vivo pharmacodynamics such as analgesia, less toxicity, higher activity, large safety window, good druggability and excellent pharmacokinetic properties.
  • the present invention also provides a method for preparing the compound of formula I and the compound of formula I-a, which is simple, easy to operate, high in yield and purity, and is suitable for industrial production.
  • the present invention also provides a salt crystal form of the compound of formula Ic-1, such as hydrochloride crystal form A, maleate crystal form B, oxalate crystal form C, and mucic acid of the compound of formula Ic-1 Salt crystal form D, fumarate crystal form E and D-glucuronate crystal form F, the salt crystal form of the compound of formula Ic-1 is in solid form, and is in phase with the free oily compound of formula Ic-1 Compared with the salt crystal form of the Ic-1 compound in solid form, it is convenient for storage, transportation and drug-making, and has high solubility and strong stability (especially excellent thermal stability and high humidity stability).
  • a salt crystal form of the compound of formula Ic-1 such as hydrochloride crystal form A, maleate crystal form B, oxalate crystal form C, and mucic acid of the compound of formula Ic-1 Salt crystal form D, fumarate crystal form E and D-glucuronate crystal form F, the salt crystal form of the compound of formula Ic-1 is in solid form, and is in phase with
  • the temperature is cooled to 25-30°C with an ice water bath.
  • Slowly add 5L of water to the system control the temperature of the system to be lower than 45°C, add 5L of ethyl acetate for extraction, and then wash the aqueous layer with 2L of ethyl acetate, combine the organic layers, and wash the organic layer with a saturated aqueous solution of sodium chloride.
  • Add 5L of water to the organic layer slowly add 0.66kg of oxalic acid under stirring conditions, continue stirring for 40 minutes, let stand for 10 minutes, add 1.70kg of 10% oxalic acid aqueous solution to the organic layer, and stir for 10 minutes.
  • the reaction system was filtered, the filter cake was washed twice with ethyl acetate, the layers were separated, the aqueous phase was washed once with ethyl acetate, the organic phases were combined, and the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain 0.73 kg oily substance target product Ic-1 compound, the yield is 54.9%, and the purity is 98.3%
  • the inhibitory activity of some of the example compounds of the present invention on the transient receptor potential channel protein TRPA1 was tested.
  • the positive control compound adopts the compound of formula A (WO2010075353):
  • Test method by IonWorks Barracuda (IWB) automated patch clamp detection HEK293 cells stably expressing TRPA1, using DMEM medium containing 15 ⁇ g/mL Blasticidin S HCl, 200 ⁇ g/mL Hygromycin B and 10% FBS serum, placed in T175 Put it in a culture flask and place it in a 37°C, 5% CO 2 incubator. When the cell density reaches ⁇ 80%, remove the culture medium, rinse it with calcium and magnesium-free phosphate buffer (PBS), and add 3 mL Trypsin was digested for 2 minutes, and 7 mL of culture medium was added to terminate the digestion.
  • IWB IonWorks Barracuda
  • Amphotericin B was freshly prepared with DMSO on the day of the experiment to make 28mg/mL, and then with intracellular fluid to make a final concentration of 0.1mg/mL.
  • the IWB experiment uses a population patch clamp (PPC) plate, and the entire detection process is automatically completed by the instrument, that is, adding extracellular fluid to the 384 wells of the PPC plate, and adding the intracellular fluid under the PPC plate, that is, into the plenum, and then adding 6L of cells Then, the intracellular fluid in plenum is replaced with amphotericin B-containing intracellular fluid, and the sealed cells are perforated to form a whole-cell recording mode.
  • the sampling frequency of recording the TPRA1 current is 10kHz
  • the cell is clamped at 0mV
  • the voltage stimulation command (channel protocol) is a ramp voltage from -100mV to +100mV in 300ms. This voltage stimulation is given every 10s, and the mTRPA1 current is induced by 300M AITC .
  • IWB IonWorks Barracuda
  • the compounds of the present invention showed potent inhibitory activity against TRPA1.
  • the half effective inhibitory concentration IC 50 of 11 compounds against TRPA1 was between 1-10 ⁇ M.
  • the half effective inhibitory concentration IC 50 of the compounds of Ic-4, Ic-8, Ic-23, and Ic-24 on TRPA1 is ⁇ 5 ⁇ M. Therefore, it can be concluded that the compound of formula I of the present invention has a strong inhibitory activity on TRPA1.
  • compound I C -10 (containing heteroaryl
  • the specific activity) and Compound C1 (phenyl containing) (compound C1 is IC 50 / Compound I IC C -10 50) is about 2.5 times, which shows that the compounds of the present invention containing a heteroaryl group (e.g., I C - 10) Higher inhibitory activity against TRPA1.
  • a heteroaryl group e.g., I C - 10
  • the A group is a compound having a naphthalene ring (e.g. duloxetine) compared to the compound I C -10 (A benzo group is an aliphatic ring) and the compounds I C -3, compounds and compound I C -23
  • the IC 50 value of I C -1 is significantly reduced.
  • S-type duloxetine the IC 50 of the compound I C -10, Compound I C -3, I or Compound C -23 the ratio of compound I IC C -1 in a compound according to any of from about 2.8 50 to 6.8 . This indicates that the compound of the present invention whose A group is a benzoalicyclic or heteroaryl group has higher inhibitory activity on TRPA1 (increased by about 2.8-6.8 times).
  • the present invention also compounds I C -1, measured by TRPA1 manual patch clamp assays inhibitory activity was measured as follows:
  • the HEK293 cell line stably expressing the human TRPA1 channel is used in DMEM medium containing 15 ⁇ g/mL Blasticidin S HCl, 200 ⁇ g/mL Hygromycin B and 10% FBS serum, placed in a T75 culture flask, and placed at 37°C. Cultivate in a %CO2 incubator. When the cell density reaches ⁇ 80%, remove the culture medium, rinse with calcium-magnesium-free phosphate buffer (PBS), add 2mL Trypsin for 2 minutes, add 8mL culture medium to stop Digestion. The cells were collected in a 15 mL centrifuge tube and centrifuged at 800 rpm for 3 minutes. After removing the supernatant, the cells were resuspended in an appropriate volume of extracellular fluid.
  • PBS calcium-magnesium-free phosphate buffer
  • Manual patch clamp detection uses HEKA system (Patch Master software) combined with EPC-10 amplifier to record the whole cell current of TRPA1 stably transfected cell line at room temperature.
  • Inner solution formula for whole cell recording 140CsCl, 10HEPES, 5EGTA, 0.1CaCl2, 1MgCl2 (pH7.2, osmotic pressure 295-300mOsm); external solution for recording uses Ca2+-free setting (mM): 140NaCl, 5KCl, 0.5 EGTA, 1MgCl2, 10Glucose, 10HEPES (pH 7.4, osmotic pressure 300-310mOsm).
  • the resistance of the glass microelectrode used for patch clamp recording is 2-4M ⁇ , the sampling frequency is 10kHz, the filter frequency is 2.9kHz, the cell is clamped at 0mV, and the voltage stimulation command (channel protocol) is a 300ms linear voltage from -100mV to +100mV, and then restored To a clamp potential of 0mV, this recording is performed every 2s.
  • the hTRPA1 current is induced by 100 ⁇ M AITC. To ensure the accuracy of the current recording, the series resistance is compensated by 60% during recording.
  • the hepatotoxicity and neurocytotoxicity experiments of the I C -10 and I C -1 compounds prepared in the example were determined by the following methods:
  • HepG-2 and SH-SY5Y cells culture them in a 10 cmdish at 37°C, 5% CO 2 in a cell incubator; trypsinize and resuspend the cells and count them. According to the 100 ⁇ l/well system, the cells are transferred to 8000cells. In a 96-well plate. Incubate in a cell incubator at 37°C and 5% CO 2 for 24 hours; prepare a compound gradient concentration system, 2-fold dilution, and the system is 100 ⁇ l/well. Remove the supernatant in the 96-well plate cell culture system on the first day, and add the newly configured drug concentration system to the culture plate wells of the cultured cells (set up double multiple wells).
  • Cytotoxicity (%) [A(0 ⁇ )-A( ⁇ )]/[A(0 ⁇ )-A (Blank)) ⁇ 100
  • Duloxetine hepatocyte toxicity and neurotoxicity were 33 ⁇ M and 28 ⁇ M, and I C-1 of the present invention and the compounds I C -10 hepatocyte toxicity and neurotoxicity (IC 50, ⁇ M) is about 60-120 ⁇ M, which indicates that the toxic and side effects of the compound of the present invention are significantly lower, and the compound is only about 1/2 or 1/3 of the toxic and side effects of duloxetine). This suggests that the compound of the present invention has excellent safety.
  • prepared by Example pain model of formalin test in mice embodiment of the present invention is a compound of I C -10, analgesic activity of the test compound, as follows:
  • mice Male, 9 weeks were taken, and the mice were randomly divided into 3 groups: solvent control group (vehicle, normal saline), duloxetine group (Duloxetine, 5-HT reuptake and NE Reuptake inhibitor) and Ic-10 group (compound Ic-10 of the present invention). Before the start of the experiment, the mice were allowed to adapt to the experimental environment for 72 hours, during which there was no need to fast and water.
  • solvent control group vehicle, normal saline
  • duloxetine group Duloxetine, 5-HT reuptake and NE Reuptake inhibitor
  • Ic-10 group compound Ic-10 of the present invention.
  • the test drug was given by intraperitoneal injection at a dose of 20 mg/kg, and then the mice were placed in a transparent, ventilated plexiglass cylinder for 1 hour, and 4% was injected with a microinjector into the left hind plantar of each group of mice 20 ⁇ l of formalin solution was used to record the foot pain response of mice in real time with a miniature camera. The number of times the mouse lifts (1 min/time), shakes (2 min/time), and licks (3 min/time) of the left foot and the length of time licking the left foot is used as an indicator of pain response. Observe and record 0-10 min (phase I). , Acute pain period) and 10-60min (Phase II, inflammatory pain period) in two periods of cumulative scoring and licking time, and statistical analysis.
  • Compound I C -10 analgesic activity according to the invention results in the formalin pain model in mice as shown in FIG. From the results, the statistical tests add enough time indicators, in the 20mg / kg dose, the compounds of this invention in phase I I C -10 (0-10 min) and phase II (10-60min) exhibited clear And the potent analgesic activity, compared with the normal saline group, almost completely inhibited the behavior of the mice due to pain caused by foot addition, and the analgesic activity of the clinical drug duloxetine is comparable.
  • mice Take SPF-grade C57 male mice, adjust the temperature of the hot plate to be constant at 55 ⁇ 0.1°C, and select mice that have painful responses such as licking their feet within 10-30 seconds (abandon those who evade and jump). If there is a pain reaction, take it out immediately to prevent the mouse from scalding.
  • the 40 selected animals were weighed, and the animals were randomly divided into 4 groups, namely the saline control group (blank control), duloxetine group (positive control group), and gabapentin group (positive control group) And group Ic-23 (compounds of the present invention).
  • test compound was freshly prepared on the day of administration.
  • a 0.9% NaCl physiological saline solution was prepared as a solvent for later use, an appropriate amount of the test compound was added to the required volume of physiological saline, and the mixture was fully suspended to prepare a drug concentration of 1 mg/ml.
  • the standard volume of administration for mice is 10ml/kg (i.e. 0.1ml/10g).
  • mice do not need to fast and water before administration.
  • the administration volume is 10ml/kg.
  • I duloxetine dose and C -23 was 10mg / kg, gabapentin dose of 100mg / kg.
  • Hot plate observation index the reaction time of mice on the hot plate at 55 ⁇ 0.1°C (Time Latency). 3h before administration and 15min, 30min, and 60min after administration, test and record each time.
  • Compound I C-23 of the present invention analgesic activity induced pain model in mice results in the hot plate 3 as shown in FIG. From the results, compared with the control group, the compounds of the present invention exhibit I C -23 at 10mg / kg dose of an analgesic effect it is very potent, with significant difference. Compared to the positive control group, in 60 minutes compounds of the invention I C -23 analgesic activity is much better than 100mg / kg gabapentin and better than 10mg / kg of the analgesic effect of duloxetine.
  • the hot plate pain model is a classic model for evaluating the efficacy of drugs for acute pain. Therefore, the compound of the present invention has an excellent therapeutic effect on acute pain.
  • the blood samples were collected on ice and centrifuged to separate the plasma (centrifugation conditions: 8000 rpm, 6 minutes, 4°C).
  • the collected plasma was stored at -80°C before analysis.
  • the linear relationship between the peak area ratio and the concentration is expressed by the correlation coefficient (R) obtained from the regression equation of the compound.
  • R correlation coefficient
  • the pharmacokinetic calculation software WinNonlin7.0 non-compartmental model was used to calculate the pharmacokinetic parameters of the test substance.
  • Internal standard working solution Pipette a certain amount of tolbutamide internal standard stock solution with a concentration of 490,000ng/mL into a volumetric flask, dilute to the mark with methanol and mix well to obtain a concentration of 200ng/mL Internal standard working solution.
  • the peak concentration (C max , 177 ng/mL) is reached at 0.083 h
  • the elimination half-life (T 1/2 ) is 1.77 h
  • the AUC (0- ⁇ ) is 449h*ng/mL
  • the peak concentration (C max , 76ng/mL) is reached at 0.83h
  • the elimination half-life (T 1/2 ) is 1.81h
  • AUC( 0- ⁇ ) is 222h*ng/mL. Calculated by AUC(0- ⁇ ), the oral bioavailability is 9.9%.
  • the compounds of the present invention having the structure of formula I (compound I C -1) compared to duloxetine has better pharmacokinetic properties, its longer half-life in the plasma exposure Higher, better bioavailability, suitable for development into oral administration, and has a good prospect of preparing medicines.
  • the experimental groups were: solvent control group, 10 mg/kg duloxetine group (positive control group) and 10 mg/kg compound Ic-1 group (the compound prepared in Example 1)
  • Duloxetine Weigh 17mg of duloxetine, dissolve it in normal saline and dilute to 8.5mL, and administer it by gavage after being fully dissolved. The volume of administration is 5ml/kg.
  • Ic-1 10mg/kg Ic-1: Weigh 17mg Ic-1, dissolve it with normal saline and make the volume to 8.5mL, and orally administer it by gavage after fully dissolving, and the volume of administration is 5ml/kg.
  • mice Take male C57BL/6 mice, weighing 18-22 grams at the beginning of the experiment, and distribute 4 mice in each cage, and they can eat and drink freely. There were 12 mice in each experimental group, and the experimental mice were labeled by the tail labeling method.
  • mice At 4:30 pm, put the mice in a plexiglass box with a stainless steel mesh, and then put the plexiglass box in a cold storage (temperature 4 ⁇ 2°C) overnight. Eat freely and use agar blocks instead of water.
  • the compound was administered orally according to the time arranged in the experiment, and the dosage was 10 mg/kg.
  • the testing method for mechanical hyperalgesia is as follows:
  • the mice will acclimate for 15 minutes before testing.
  • use the test fiber to test the center of the left hind foot of the mouse.
  • the test fiber includes 8 test strengths: 2.36(0.02g), 2.44(0.04g), 2.83(0.07g), 3.22(0.16g), 3.61(0.4g), 3.84(0.6g), 4.08(1g), 4.17 (1.4g).
  • the animal's rapid withdrawal of its feet was recorded as a pain response.
  • the withdrawal of the animal's feet when the test fiber leaves the animal's skin is also recorded as a pain response. If the animal moves or walks, the pain response is not remembered, and the test should be repeated. In the test, first use 3.22 (0.16g). If the animal has a pain response, use the test fiber with a lower strength in the next test; if the animal does not respond to pain, use the test fiber with a higher strength (Chaplan et al. 1994). The maximum strength of the tested fiber is 4.17 (1.4g).
  • test results are recorded in Table 4 below, with pain response record X and no pain response record O.
  • the compound Ic-1 of the present invention exhibits a very potent analgesic effect at a dose of 10 mg/kg, and it is shown to inhibit ICS after 1 hour and 2 hours of oral administration. Mechanical hyperalgesia caused by the model. Compared with the positive control group, compound Ic-1 has better analgesic effects than duloxetine at the three test times of 0.5h, 1h and 2h.
  • the mouse ICS model is a classic model for evaluating the efficacy of drugs in the treatment of fibromyalgia. Therefore, the compound Ic-1 of the present invention has an excellent therapeutic effect on fibromyalgia.
  • ICR mice Take ICR mice, male, 22-25g, fast for 2h before administration, and can't help water. All ICR mice were weighed and grouped randomly, with the number of animals in each group >10.
  • the negative control group is a normal saline group (Vehicle, blank control), and the positive control group is set to a dose of 10 mg/kg indomethacin (a non-steroidal anti-inflammatory drug) and a dose of 10 mg/kg anisodamine (An antispasmodic drug with clinically analgesic activity)
  • Duloxetine was administered at doses of 10 mg/kg and 20 mg/kg.
  • the test compound was I C -1 (the compound prepared in Example 1), and the administration dose was set at 5 mg/kg and 10 mg/kg.
  • mice According to the weight of the mouse, it was administered by gavage. Inject 1.5% acetic acid solution (0.1ml/10g) into the intraperitoneal cavity 1 hour after administration, and observe the number of times of visceral pain in each group of mice within 30 minutes. The mice have concave abdomen, the trunk and hind legs are stretched, and the buttocks are high. Once, finally count the number of occurrences of the above phenomenon within 30 minutes. After administration, the fewer visceral pains in mice, the stronger the analgesic effect of the compound.
  • acetic acid solution 0.1ml/10g
  • the animals were acclimatized in the experimental environment for 15 minutes/day for 3 days.
  • the rats were subjected to a baseline test of mechanical hyperalgesia, and animals that did not exhibit mechanical hyperalgesia (with the withdrawal threshold greater than 5g) were eliminated and randomly divided into 1 control group and 2 experimental groups.
  • the test fiber includes 8 test strengths: 3.61 (0.4g), 3.84 (0.6g), 4.08 (1g), 4.31 (2g), 4.56 (4g), 4.74 (6g), 4.93 (8g), 5.18 (15g).
  • the animal's rapid withdrawal of its feet was recorded as a pain response.
  • the withdrawal of the animal's feet when the test fiber leaves the animal's skin is also recorded as a pain response. If the animal moves or walks, the pain response is not remembered, and the test should be repeated.
  • 4.31 (2g) If the animal has a pain response, use the test fiber with a lower strength in the next test; if the animal has no pain response, use the test fiber with a higher strength in the next test).
  • the maximum strength of the tested fiber is 5.18 (15g).
  • PWT withdrawal threshold
  • the compounds of the present invention I C -1 exhibited very potent analgesic effect at 10mg / kg dose, with a significant difference.
  • the present invention is one hour analgesic activity of the compounds I C-1 and 100mg / kg gabapentin considerable analgesic effect after administration.
  • Rat SNL model efficacy is evaluated for classical drug treatment of neuropathic pain, the compounds of the present invention, therefore I C -1 has an excellent therapeutic effect on neuropathic pain.
  • mice Male, 9 weeks
  • mice were randomly divided into 10 groups, used for the analgesic activity test of 2 compounds in the mouse formalin pain model: respectively
  • the duloxetine group is the duloxetine group and the compound I C -1 group (the compound prepared in Example 1).
  • the mice were allowed to adapt to the experimental environment for 72 hours, during which there was no need to fast and water.
  • the test drug was given by intraperitoneal injection, and the dosage settings were as follows:
  • Duloxetine group blank Vehicle (blank saline control), 1mg/kg, 5mg/kg, 10mg/kg and 20mg/kg;
  • Compound I C -1 group Blank Vehicle (blank normal saline control, isoduloxetine blank group), 0.1 mg/kg, 0.5 mg/kg, 1 mg/kg, 5 mg/kg and 10 mg/kg.
  • ED 50 refers to the dose of the drug that reduces the time to fill up by half compared with the blank control group. The smaller the ED 50 value, the lower the effective dose of the analgesic compound, the better analgesic effect.
  • Table. 7 The statistical results of the addition time of compound Ic-1 and duloxetine in the phase II (10-60min) phase at different doses in the mouse formalin model
  • the filling time of phase II (10-60 min) of the compound Ic-1 of the present invention has been reduced by more than 50% compared with that of blank Vehicle.
  • the ED 50 of analgesic effect in phase II pain is 2.22 mg/kg, while the ED 50 of duloxetine in phase II pain is 8.00 mg/kg.
  • the analgesic activity of the compound Ic-1 of the present invention is significantly better than that of duloxetine.
  • the compound Ic-1 of the present invention exhibits extremely strong analgesic activity in a mouse formalin pain model.
  • the mouse formalin model is a classic model for evaluating the efficacy of drugs for acute pain and inflammatory pain. Therefore, the compound Ic-1 of the present invention has an excellent therapeutic effect on acute pain and inflammatory pain.
  • XRPD X-ray powder diffraction
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • DVS dynamic moisture adsorption
  • X-ray powder diffraction analysis method PANalytical X-ray powder diffraction analyzer, working voltage: 45kV, working Current: 40mA, using Cu target to obtain X-ray powder diffraction pattern.
  • DSC Differential scanning calorimetry
  • Thermogravimetric analysis (TGA) analysis the instrument is TA Q5000/Discovery TGA5500; scanning speed: 10°C/min; protective gas, nitrogen.
  • DVS Intrinsic produced by SMS (Surface Measurement Systems); temperature, 25°C; carrier gas, flow rate: nitrogen, 200 ml/min; mass change per unit time: 0.002%/min; relative Humidity range: 0%RH-95%RH.
  • Nuclear magnetic analysis The instrument is a Bruker 400M nuclear magnetic resonance instrument.
  • HPLC High performance liquid chromatography purity
  • Ion Chromatography test counter ions to determine the molar ratio collected by Thermo ICS1100.
  • the compound of formula I C -1 prepared in Example 1 is an oily substance.
  • the inventors adopted a method of suspension and stirring Tested hydrochloric acid, sulfuric acid, L-aspartic acid, maleic acid, L-pyroglutamic acid, phosphoric acid, L-glutamic acid, mucic acid, L-(+)-tartaric acid, fumaric acid, citric acid, D-glucuronic acid, L-(-)-malic acid, hippuric acid, D-gluconic acid, glycolic acid, lactic acid, L-ascorbic acid, succinic acid, adipic acid, lauric acid, methanesulfonic acid, oxalic acid, malonic acid
  • the salt formation of acid, benzoic acid, S-(+)-mandelic acid and the compound of formula Ic-1 shows that the salt formed by most acid ligands is
  • the free base of the compound of formula Ic-1 is used as a starting material to obtain other salt forms.
  • the free base of the compound of formula Ic-1 is an oily substance.
  • the solubility of the free base of the compound of formula Ic-1 in 11 common solvents under room temperature conditions was tested. In the experiment, weigh about 2 mg of the Ic-1 compound oily sample into a 3-ml vial, and then gradually (50, 50, 200, 700 microliters) of the corresponding solvent was added and shaken until the solution was clear. If the sample is still not dissolved after adding the solvent to 1 ml, no more solvent is added.
  • the rough solubility range calculated based on the mass of the solid sample, the volume of the added solvent and the observed dissolution phenomenon is shown in Table 8. This data provides a reference for the selection of solvents in the screening experiment.
  • the X-ray powder diffraction data of the hydrochloride salt crystal form A of the compound of formula Ic-1 obtained in this example is shown in Table 9, the XRPD pattern is shown in Fig. 8, and the TGA/DSC superimposed pattern is shown in Fig. 9.
  • the X-ray powder diffraction data of the maleate salt crystal form B of the compound of formula Ic-1 obtained in this example is shown in Table 10, the XRPD pattern is shown in Fig. 11, and the TGA/DSC superimposed pattern is shown in Fig. 12.
  • the differential scanning calorimetry chart shows that the maleate salt crystal form B of the compound of formula Ic-1 begins to show an endothermic peak when heated to around 105.8°C.
  • the thermogravimetric analysis chart shows that the crystal form B is in There is a weight loss of about 0.8% when heated to 80°C.
  • the 1 H NMR spectrum of the maleate salt crystal form B of the compound of formula Ic-1 is shown in FIG. 13, and the result shows that the molar ratio of free base/acid is 1:1.
  • the XRPD test results of maleate crystal form B at variable temperature are shown in Figure 14. After the maleate crystal form B was purged with nitrogen and heated to 80° C. under the protection of nitrogen, no change in the crystal form was observed, indicating that the maleate crystal form B was an amorphous form.
  • the X-ray powder diffraction data of the oxalate salt crystal form C of the compound Ic-1 obtained in this example is shown in Table 11, the XRPD pattern is shown in Fig. 15, and the TGA/DSC overlay pattern is shown in Fig. 16.
  • HPLC/IC results showed that the molar ratio of free base/acid in the sample was 1:1.
  • the XRPD test results of oxalate crystal form C at variable temperature are shown in Figure 17.
  • the oxalate crystal form C was purged with nitrogen and heated to 100°C under the protection of nitrogen, and no change in crystal form was observed.
  • the X-ray powder diffraction data of the mucic acid salt crystal form D of the compound of formula Ic-1 obtained in this example is shown in Table 12, the XRPD pattern is shown in Fig. 18, and the TGA/DSC overlay pattern is shown in Fig. 19.
  • the 1 H NMR spectrum of the crystalline form D of the mucic acid salt of the compound of formula Ic-1 is shown in FIG. 20, and the result shows that the molar ratio of the free state to the mucic acid in the sample is 2:1.
  • the X-ray powder diffraction data of the crystal form E obtained in this example is shown in Table 13, the XRPD pattern is shown in Fig. 21, and the TGA/DSC superimposed pattern is shown in Fig. 22.
  • the 1 H NMR spectrum of the fumarate salt crystal form E of the compound of formula Ic-1 is shown in FIG. 23, and the result shows that the molar ratio of the free state to the fumaric acid in the sample is 1:1.
  • the X-ray powder diffraction data of the crystal form F obtained in this example is shown in Table 14, the XRPD pattern is shown in Fig. 24, and the TGA/DSC overlay pattern is shown in Fig. 25.
  • This example examines the characteristics of the salt crystal form of the compound of formula Ic-1 prepared in Example 23.
  • a dynamic moisture adsorption (DVS) instrument was used to test its hygroscopicity. The results are shown in Figure 26 and Figure 28.
  • the weight change of the hydrochloride crystal form A and maleic acid crystal form B is less than 0.2% under the conditions of 25°C/80RH, indicating that There is no hygroscopicity, indicating that the salt type is easy to store and does not need to be stored in dry conditions (refer to the 2015 edition of the Chinese Pharmacopoeia (Guidelines for Drug Hygroscopicity Test)).
  • the compound of formula Ic-1 hydrochloride crystal form A and maleate crystal form B use pH1.9SGF (simulated gastric juice), pH6.5FaSSIF (simulated artificial intestinal fluid under fasting state), pH 5.0FeSSIF (simulated artificial intestinal fluid under fed state), respectively Intestinal juice) and water were prepared into a saturated solution, and the dynamic solubility at 37°C was tested.
  • the initial dosage of hydrochloride crystal form A in H 2 O, SGF, and FaSSIF was about 20 mg/mL
  • the initial dosage in FeSSIF was about 40 mg/mL.
  • the initial dosage of maleate crystalline form B in H 2 O, SGF, FaSSIF and FeSSIF is about 10 mg/mL.
  • hydrochloride crystal form A and maleate crystal form B of the compound of formula Ic-1 have excellent solubility, and the solubility in water is also significantly higher than that of the free form of Ic-1 compound, which is beneficial to improve the drug Absorption and medicinal properties.
  • the hydrochloride crystal form A of the compound of formula Ic-1 was tested for its equilibrium solubility in water for 24 hours at room temperature.
  • the material was fed with a solid dosage of ⁇ 300mg/mL, and the suspension was obtained after 24 hours of magnetic stirring (750rpm) in water. After centrifugation, the supernatant was filtered through a 0.22 micron PTFE filter membrane and then subjected to HPLC test, and the obtained solid was subjected to XRPD test. The results showed that the solid crystal form did not change after the equilibrium solubility test.
  • the hydrochloride crystal form A has a solubility of 210.5 mg/mL in water (free state concentration, converted to a hydrochloride concentration of 237.3 mg/mL).
  • the SADABS program is used to perform empirical absorption correction on the data
  • SHELXT2014 is used to analyze the single crystal structure through the direct method, and the structure is refined by the least square method.
  • the hydrogen atom refinement process takes all directions It is obtained by the same calculation process.
  • the hydrogen atom on N is obtained by the residual electron density, and the hydrogen atom on CH is obtained by calculation and hydrogenation, and the riding model is adopted to refine it.
  • the Flack constant is -0.09(5), and the chiral center C4 of the hydrochloride salt crystal form A of the compound of formula Ic-1 prepared in Example 23 can be confirmed as the S configuration (as shown in Figure 32).
  • TRPM8 cells were cultured in a cell culture dish with a diameter of 35mm, placed in a 37°C, 5% CO2 incubator, and subcultured at a ratio of 1:5 every 48 hours.
  • DMEM Invitrogen
  • 2mM L-glutamine 50 ⁇ g/mL Hygromycin B (Invitrogen)
  • 5 ⁇ g/mL Blasticidine HCl Invitrogen
  • TRPV1 cells were cultured in a cell culture dish with a diameter of 35mm, placed in a 37°C, 5% CO2 incubator, and subcultured at a ratio of 1:5 every 48 hours.
  • DMEM Invitrogen
  • 2mM L-glutamine 50 ⁇ g/mL Hygromycin B (Invitrogen)
  • 5 ⁇ g/mL Blasticidine HCl Invitrogen.
  • TRPV4 cells were cultured in a cell culture dish with a diameter of 35mm, placed in a 37°C, 5% CO2 incubator, and subcultured at a ratio of 1:5 every 48 hours.
  • DMEM Invitrogen
  • 2mM L-glutamine 50 ⁇ g/mL Hygromycin B (Invitrogen)
  • 5 ⁇ g/mL Blasticidine HCl Invitrogen
  • Menthol preparation Take 100.00mM Menthol DMSO mother solution and add it to the extracellular fluid, and dilute it 1000 times to 100.00 ⁇ M Menthol solution.
  • AMTB preparation Take 5 ⁇ L of 20.00mM AMTB DMSO mother solution and add it to 4995 ⁇ L of 100.00 ⁇ M Menthol solution, and dilute it by 1000 times to obtain the final concentration of 20.00 ⁇ M AMTB that needs to be tested.
  • Capsaicin preparation Take 1.00mM Capsaicin DMSO mother solution and add it to the extracellular fluid, and dilute it by 1000 times to 1.00 ⁇ M Capsaicin solution.
  • Capsazepine preparation Take 5 ⁇ L of 10.00mM Capsazepine DMSO mother solution and add it to 4995 ⁇ L of 1.00 ⁇ M Capsaicin solution, and dilute it by 1000 times to obtain the final concentration of 10.00 ⁇ M Capsazepine solution that needs to be tested.
  • GSK1016790A preparation Take 30.00 ⁇ M GSK1016790A DMSO mother solution and add it to the extracellular fluid, and dilute it by 1000 times to 30.00nM GSK1016790A solution.
  • RR (Ruthenium Red) preparation Take 20 ⁇ L of 10.00mM RR ddH2O mother solution and add it to 4980 ⁇ L of 30.00nM GSK1016790A solution, and dilute 250 times to obtain the final concentration of 40.00 ⁇ M RR solution to be tested.
  • Ic-1 preparation Take 20.00mM or 10.00mM Ic-1 mother solution and add it to the agonist solution of the corresponding channel, and dilute it by 1000 or 2000 times to the final concentration of 10.00M LDS solution to be tested.
  • the content of DMSO in the final test concentration does not exceed 0.2%, and this concentration of DMSO has no effect on the channel current.
  • the channel currents induced by the corresponding agonists were recorded by the whole-cell patch clamp technique at room temperature.
  • the glass microelectrode is made of a glass electrode blank (BF150-86-10, Sutter) drawn by a drawing instrument (P97, Sutter). The tip resistance after filling the electrode liquid is about 2-5M ⁇ . Insert the glass microelectrode The amplifier probe can then be connected to the patch clamp amplifier. The clamping voltage and data recording are controlled and recorded by the pClamp 11 software through a computer.
  • the sampling frequency is 10kHz and the filtering frequency is 2kHz.
  • the cells were clamped at 0mV, and the 300ms ramp voltage was from -100mV to +100mV. This voltage was applied every 2s and an agonist was given to induce channel currents. The current was induced for more than 40s and no attenuation was confirmed before the drug administration process was started. Give at least 20s at each test concentration, and test at least 3 cells at each concentration (n ⁇ 3).
  • Inhibition% represents the percentage of the compound's inhibition of current
  • I and Io represent the amplitude of the current after and before drug addition, respectively.
  • the inhibitory percentages of the compound of formula I C -1 on the currents of the three channels at a concentration of 10.00 ⁇ M were 41.07%, 22.48% and 4.68%, respectively (inhibition percentages less than 50% indicate The IC50 of the compound to TRPM8 is greater than 10 ⁇ M), and the corresponding positive compounds AMTB, Capsazepine and RR have an inhibition percentage of more than 90%. From the above results, it can be seen that compound Ic-1 is a specific inhibitor of TRPA1.

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Abstract

提供式I化合物、其制备方法、盐酸盐晶型A。所述的式I化合物、其盐酸盐晶型A对瞬时受体电位通道蛋白(TPR)具有优异的抑制作用，并对瞬时受体电位通道蛋白相关的疾病具有良好的治疗作用。

Description

3-芳氧基-3-五元杂芳基-丙胺类化合物的制备方法及晶型 技术领域
本发明涉及药物化学和药物治疗学领域，具体地涉及一种3-芳氧基-3-五元杂芳基-丙胺类化合物的制备方法及晶型。
背景技术
疼痛被称为第五大生命体征，是机体组织受到损害的警戒信号。疼痛是患者就医的最常见原因之一，根据持续时间分为急性疼痛(急剧发病，持续时间短，也可呈持续状态)和慢性疼痛(发病缓慢或由急性疼痛转化而来，持续时间长，亦可呈间断发作，很多慢性疼痛查不出明显的损伤)。急性疼痛多是由组织创伤引起的伤害性疼痛，包括手术后疼痛，创伤、烧伤后疼痛，分娩痛，心绞痛、胆绞痛、肾绞痛等内脏痛，骨折痛，牙痛，癌性疼痛等。手术和创伤后疼痛是临床最常见和最急需处理的急性疼痛。慢性疼痛主要包括神经病理性疼痛、痛骨关节炎、慢性腰背痛和血管源性痛等。三叉神经痛、糖尿病性疼痛、坐骨神经痛或带状孢疹后神经痛为神经病理性疼痛的主要类型。神经病理性疼痛全球患病率大约是10％，发病率高，患者群体数量多。在美国有10％-30％的人患有慢性疼痛，造成每年约6350亿美元的社会支出，超过癌症和心脏病的总和。慢性疼痛病因复杂，属难治性疾病，只有不足50％的患者能够通过药物治疗达到有效的镇痛。预计2026年中国神经痛药物总市场规模接近260亿元，离子通道类神经痛药物市场规模超200亿元以上。
传统镇痛药物主要包括阿片类药物和非甾体抗炎药。阿片类药物镇痛作用强，但长期使用易导致耐受性、依赖性和成瘾性，并且有呼吸抑制、中枢镇静等不良反应。非甾体抗炎药仅发挥中等程度镇痛作用，同时具有消化道出血和心脏毒性等反应。近日美国国家安委会发布关于可预防死亡报告显示，美国历史上第一次，阿片类药物过量致死超过了车祸所致死亡人口比例。根据委员会对2017年意外死亡数据的分析，每96名美国人中，就有1名死于阿片类药物过量，而车祸致死的数据为每103名美国人中有1名。阿片类药物滥用已经造成了如今席卷全美国的严重社会危机，因此市场需要新机制的镇痛药物。
TRPA1是TRP离子通道超家族中的一员，TRPA亚家族的唯一成员，属于非选择性阳离子通道，可通透Na +，K +，Ca 2+和Mg 2+。TRPA1主要分布在背根神经(DRG)、三叉神经(TG)和迷走神经(VG)的初级感觉神经元上。从分布的人体系统来看，TRPA1高表达在外周神经系统、呼吸系统、胃肠系统和泌尿系统，当这些器官组织出现功能异常时，TRPA1通道的表达和功能通常也同步发生异常。TRPA1可以将冷刺激、化学刺激以及机械刺激转化为内向电流，引发一系列生理功能，并参与多种痛觉的形成。炎性痛是某些慢性疾病的共同困扰，临床上尚缺乏很有效的治疗手段。动物实验研究显示，TRPA1参与炎性反应，并在炎性疼痛中发挥重要作用，通过使用TRPA1特异性阻断剂，可以明显减轻大鼠炎性疼痛反应。从目前的研究来看，TRPA1在哮喘和咳嗽的发生中扮演重要的作用，诱导哮喘与咳嗽的化合物，无论是细胞内源因子，还是外源因子，都能激活TRPA1。TRPA1的拮抗剂能够减轻哮喘症状，能阻断气道高反应性。通过不同的内脏高敏感动物模型如结肠炎、直结肠扩张或应激，证实TRPA1参与内脏高敏感的调控，在内脏痛中发挥重要作用。神经源性疼痛是由中枢或外周神经系统损伤或者疾病引起的疼痛综合征，主要表现为痛觉过敏、异常痛敏和自发性疼痛等。近年来越来越多的研究显示，TRPA1通道在不同的神经源性疼痛中起到重要作用，例如糖尿病性神经病变和化疗药引起的神经病变等。最近研究还表明，TRPA1在牙痛、偏头痛等疼痛中也有介导作用，通过给予TRPA1的拮抗剂能明显缓解疼痛 症状的产生。
TRPA1在人体系统中广泛分布和表达，除以上TRPA1参与的生理功能外，目前已报道的TRPA1抑制剂适应症的开发还涉及到炎症性肠病、慢性阻塞性肺疾病、镇咳、止痒、过敏性鼻炎、耳疾病、抗糖尿病、尿失禁等。TRPA1是已经被证实了的多种疾病治疗的新靶点。
因此，考虑到目前疼痛治疗作为临床未满足的需求，以及目前已有治疗药物的存在的诸多问题，本领域迫切需要开发一种针对TRP靶点(尤其是TRPA1靶点)的治疗药物，从而提高疾病的治疗效果。
发明内容
本发明的目的在于提供一种结构新颖的以TRP通道为靶点(尤其是TRPA1靶点)的化合物，及其制备方法、晶型和用途。
本发明第一方面，提供一种式I化合物的制备方法，所述的方法包括步骤：
(1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式a化合物与式b化合物反应，得到式I化合物；`
Figure PCTCN2021076337-appb-000001
其中：
环A为取代或未取代的4-12元碳环、取代或未取代的4-12元杂环、取代或未取代的5-12元杂芳环、取代或未取代的C 6-C 12芳环；
R 1和R 2各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
X和Y各自独立地为碳原子、氧原子、硫原子或氮原子；
R 3为氢、卤素、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
R 4为卤素；
R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基，取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
W为O或S；
n为1、2或3；
其中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、C 6-C 12芳基、5-10元杂芳基；
其中，所述的杂环、杂芳环和杂芳基各自独立地具有1-3个(优选为1、2或3个)选自N、O和S杂原子。
在另一优选例中，环A为取代或未取代的5-10元碳环、取代或未取代的5-10元杂环、取代或未取代的5-12元杂芳环、取代或未取代的C 6-C 12芳环。
在另一优选例中，环A为取代或未取代的5-10元碳环、取代或未取代的5-10元杂环、取代或未取代的5-12元杂芳环。
在另一优选例中，环A不为苯环。
在另一优选例中，环A为取代或未取代的5-7元碳环、取代或未取代的5-7元杂环、取代或未取代的5-7元杂芳环。
在另一优选例中，环A为取代或未取代的5-7元碳环或5-7元杂芳环。
在另一优选例中，环A为取代或未取代的5元碳环、取代或未取代的6元碳环或取代或未取代的呋喃环。
在另一优选例中，环A为5元碳环、6元碳环或呋喃环。
在另一优选例中，环A为
Figure PCTCN2021076337-appb-000002
在另一优选例中，环A与相邻苯环的连接结构为：
Figure PCTCN2021076337-appb-000003
在另一优选例中，X和Y中至少一个是为杂原子。
在另一优选例中，Y为碳原子或氮原子。
在另一优选例中，X为S或O。
在另一优选例中，X为S。
在另一优选例中，Y为碳原子。
在另一优选例中，R 1和R 2各自独立为氢、取代或未取代的C 1-C 8烷基、取代或未取代的C 3-C 8环烷基、取代或未取代的5-10元杂芳基、取代或未取代的C 6-C 12芳基。
在另一优选例中，R 1和R 2各自独立为氢、取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基。
在另一优选例中，R 1和R 2各自独立为氢或取代或未取代C 1-C 3烷基。
在另一优选例中，R 1和R 2各自独立为氢、甲基或乙基。
在另一优选例中，R 3为氢、卤素、取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 10环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基。
在另一优选例中，R 3为氢、卤素、取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基。
在另一优选例中，R 3为氢、卤素、取代或未取代的C 1-C 6烷基。
在另一优选例中，R 3为氢、卤素、取代或未取代的C 1-C 4烷基。
在另一优选例中，R 3为氢原子、氯原子或甲基。
在另一优选例中，所述的卤素为F、Cl、Br或I。
在另一优选例中，R 4为F、Cl、Br或I。
在另一优选例中，R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 10烷基、取代或未取代的C 3-C 8环烷基，取代或未取代的5-10元杂芳基、取代或未取代的C 6-C 12芳基。
在另一优选例中，R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基。
在另一优选例中，R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢。
在另一优选例中，W为O或S。
在另一优选例中，W为O。
在另一优选例中，n为1或2。
在另一优选例中，n为1。
在另一优选例中，n≥2时，各个R 3相同或不同。
在另一优选例中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、六元的芳基、五元或六元的杂芳基(优选C 5杂芳基)。
在另一优选例中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、羟基、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、六元的芳基、五元或六元的杂芳基(优选C 5杂芳基)。
在另一优选例中，所述的
Figure PCTCN2021076337-appb-000004
结构为：
Figure PCTCN2021076337-appb-000005
在另一优选例中，所述的式a化合物为具有式a-1结构：
Figure PCTCN2021076337-appb-000006
在另一优选例中，所述的式b化合物为具有式b-1结构：
Figure PCTCN2021076337-appb-000007
在另一优选例中，所述的式I化合物为具有式I-A结构：
Figure PCTCN2021076337-appb-000008
在另一优选例中，所述的式I化合物选自下组：
Figure PCTCN2021076337-appb-000009
在另一优选例中，所述的式I化合物选自下组：
Figure PCTCN2021076337-appb-000010
在另一优选例中，所述的第一溶剂选自下组：二甲基亚砜、甲苯、DMF，或其组合。
在另一优选例中，所述的第一溶剂选自下组：二甲基亚砜。
在另一优选例中，所述的第一催化剂选自下组：卤化盐、四丁基溴化铵、4-二甲 氨基吡啶、二苯并18冠醚6、吡啶，或其组合。
在另一优选例中，所述的卤化盐选自下组：氟化钾、氯化钾、溴化钾、碘化钾、氟化钠、氯化钠、溴化钠、碘化钠，或其组合。
在另一优选例中，所述的第一催化剂包括碘化钾。
在另一优选例中，所述的第一碱试剂选自下组：无机碱、有机碱，或其组合。
在另一优选例中，所述的第一碱试剂选自下组：钠氢、氢氧化物，或其组合。
在另一优选例中，所述的氢氧化物选自下组：氢氧化钠、氢氧化钾，或其组合。
在另一优选例中，所述的第一碱试剂包括氢氧化物。
在另一优选例中，所述的第一碱试剂包括氢氧化钠。
在另一优选例中，所述的步骤(1)中，所述反应的温度为30-90℃，较佳地30-80℃，更佳地40-70℃，最佳地50～60℃。
在另一优选例中，所述的步骤(1)中，所述反应的时间为10-48h，较佳地16-40h，更佳地20-35h，最佳地22-25h。
在另一优选例中，式a化合物与式b化合物的摩尔比为1:1-3，较佳地1:1-2，更佳地1:1.2-1.7。
在另一优选例中，式a化合物与第一碱试剂的摩尔比为1：2.5～10，较佳地1:3-8，更佳地1:4-6。
在另一优选例中，式b化合物与第一碱试剂的摩尔比为1～3：2.5～10，较佳地1-2:3-8，更佳地1.2-1.7:4-6。
在另一优选例中，所述的第一催化剂与所述的第一碱试剂的摩尔比为1:5-70，较佳地1:10-60，更佳地1:15-50，更佳地1:20-50，更佳地1:25-40，最佳地1:28-36。
在另一优选例中，所述的第一催化剂与所述的式a化合物的摩尔比为1:1-50，较佳地1:2-40，更佳地1:3-30，更佳地1:5-25，更佳地1:5-20，最佳地1:5-15。
在另一优选例中，所述的步骤(1)中，所述的反应在常压下(如1atm)下进行。
在另一优选例中，所述的步骤(1)中，所述的反应在N 2保护下进行。
在另一优选例中，所述的步骤(1)包括步骤：
将式a化合物溶于第一溶剂中，加入式b化合物和第一催化剂，降温后加入第一碱试剂，升温反应得到式I化合物。
在另一优选例中，所述降温后的温度为5～30℃(较佳地10～20℃)。
在另一优选例中，所述升温后的温度为30-90℃，较佳地30-80℃，更佳地40-70℃，最佳地50～60℃。
在另一优选例中，所述的步骤(1)中，反应结束后，向反应液中加入水和乙酸乙酯进行萃取，收集有机相，分离得到式I化合物。
在另一优选例中，所述的有机相将酸水溶液洗涤后，分离得到式I化合物。
在另一优选例中，所述的酸水溶液为草酸水溶液。
本发明第二方面，提供一种式I-1化合物的制备方法，所述的方法包括步骤：
(1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式a化合物与式b化合物反应，得到式I化合物；
Figure PCTCN2021076337-appb-000011
(2)在第二溶剂中，在第二碱试剂的存在下，式I化合物与式c所示的N-去甲基化试剂反应，得到式I-a化合物；
Figure PCTCN2021076337-appb-000012
(3)在第三溶剂中，在第三碱试剂的存在下，式I-a化合物发生水解反应，得到式I-1化合物；
Figure PCTCN2021076337-appb-000013
其中，
环A为取代或未取代的4-12元碳环、取代或未取代的4-12元杂环、取代或未取代的5-12元杂芳环、取代或未取代的C 6-C 12芳环；
R 1和R 2各自独立为取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基；
X和Y各自独立地为碳原子、氧原子、硫原子或氮原子；
R 3为氢、卤素、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
R 4为卤素；
R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基，取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
R 12为取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的C 6-C 16芳基、取代或未取代的5-16元杂芳基、取代或未取代的C 1-C 6烷基-W-、取代或未取代的C 3-C 12环烷基-W-、取代或未取代的C 6-C 16芳基-W-、取代或未取代的5-16元杂芳基-W-；
R 13为卤素；
W为O或S；
n为1、2或3；
其中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、C 6-C 12芳基、5-10元杂芳基；
其中，所述的杂环、杂芳环和杂芳基各自独立地具有1-3个(优选为1、2或3个)选自N、O和S杂原子。
在另一优选例中，环A、X、Y、W、n、R 1、R 2、R 3、R 4、R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立地如本发明第一方面所述。
在另一优选例中，所述的第一溶剂中、在第一催化剂和第一碱试剂各自独立地如本发明第一方面所述。
在另一优选例中，式a化合物、式b化合物和式I化合物各自独立地如本发明第一方面所述。
在另一优选例中，所述的步骤(1)如本发明第一方面所述。
在另一优选例中，所述的第二溶剂选自下组：二甲基亚砜、甲苯，或DMF，或其组合。
在另一优选例中，所述的第二溶剂为甲苯。
在另一优选例中，所述的第二碱试剂选自下组：三乙醇胺(TEA)、1，8-二氮杂二环十一碳-7-烯(DBU)、N,N-二异丙基乙胺(DIEA)、碳酸钠，或其组合。
在另一优选例中，所述的第二碱试剂包括N,N-二异丙基乙胺(DIEA)。
在另一优选例中，所述的步骤(2)中，所述反应的温度为20-80℃，较佳地30-70℃，更佳地30-60℃，更佳地40-50℃，最佳地40～45℃。
在另一优选例中，所述的步骤(2)中，所述反应的时间为2-16h，较佳地2-10h，更佳地2-6h，最佳地3-5h。
在另一优选例中，所述的式c所示的N-去甲基化试剂包括氯甲酸苯酯。
在另一优选例中，在所述的步骤(2)中，所述的式I化合物与第二碱试剂的摩尔比为1:0.2-10，较佳地1:0.5-8，更佳地1:0.8-5，更佳地1:1-3，最佳地1:1-2。
在另一优选例中，在所述的步骤(2)中，所述的式I化合物与所述的式c化合物的摩尔比为1:0.2-10，较佳地1:0.5-8，更佳地1:0.8-5，更佳地1:1-3，更佳地1:1.1～2.5，最佳地1:1-2。
在另一优选例中，在所述的步骤(2)中，所述的第二碱试剂与所述的式c化合物的摩尔比为0.8-5：0.8-5，较佳地1-3：1-3，更佳地1～2：1.1～2.5，最佳地1～2：1.1～2。
在另一优选例中，所述的步骤(2)中，所述的反应在常压下(如1atm)下进行。
在另一优选例中，所述的步骤(2)中，所述的反应在N 2保护下进行。
在另一优选例中，所述的步骤(2)包括步骤：
将式I化合物和第二碱试剂溶于第二溶液中，降温后加入式c所示的N-去甲基化试剂，升温反应得到式I-a化合物。
在另一优选例中，所述降温后的温度为10～40℃，较佳地20～30℃。
在另一优选例中，所述升温后的温度为20-80℃，较佳地30-70℃，更佳地30-60℃，更佳地40-50℃，最佳地40～45℃。
在另一优选例中，所述的步骤(2)中，反应结束后，向反应液中加入水和乙酸乙酯萃取，收集有机相，浓缩得到式I-a化合物。
在另一优选例中，所述的式I-a化合物具有式I-a-1的结构：
Figure PCTCN2021076337-appb-000014
在另一优选例中，所述的第三溶剂选自下组：二甲基亚砜、甲苯，或DMF，或其组合。
在另一优选例中，所述的第三溶剂为二甲基亚砜。
在另一优选例中，所述的第三碱试剂选自下组：无机碱、有机碱，或其组合。
在另一优选例中，所述的第三碱试剂选自下组：叔丁醇钾、碳酸钾、氢氧化物，或其组合。
在另一优选例中，所述的氢氧化物选自下组：氢氧化钠、氢氧化钾，或其组合。
在另一优选例中，所述的第三碱试剂包括氢氧化物。
在另一优选例中，所述的第三碱试剂包括氢氧化钠。
在另一优选例中，所述的步骤(3)中，所述反应的温度为20-100℃，较佳地30-90℃，更佳地40-80℃，更佳地50-70℃，最佳地60～65℃。
在另一优选例中，所述的步骤(3)中，所述反应的时间为5-16h，较佳地5-10h，更佳地7-10h。
在另一优选例中，所述的步骤(3)中，所述的式I-a化合物与第三碱试剂的摩尔比为1:1-20，较佳地1:2-15，更佳地1:2-10，更佳地1：3.5～8，最佳地1:6。
在另一优选例中，所述的步骤(3)中，反应结束后，向反应液中加入水和乙酸乙酯进行萃取，收集有机相，分离得到式I化合物。
在另一优选例中，所述的有机相将酸水溶液洗涤后，分离得到式I化合物。
在另一优选例中，所述的酸水溶液为草酸水溶液。
在另一优选例中，所述的式I-1化合物具有式I-1-1的结构：
Figure PCTCN2021076337-appb-000015
在另一优选例中，所述的方法包括步骤：
(1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式i化合物与式ii化合物反应，得到式Ic-8化合物；
Figure PCTCN2021076337-appb-000016
(2)在第二溶剂中，在第二碱试剂的存在下，式Ic-化合物与氯甲酸苯酯反应，得到式iii化合物；
Figure PCTCN2021076337-appb-000017
(3)在第三溶剂中，在第三碱试剂的存在下，式iii化合物发生水解反应，得到式Ic-1化合物；
Figure PCTCN2021076337-appb-000018
本发明第三方面，提供一种式I-a化合物的制备方法，所述的方法包括步骤：
(2)在第二溶剂中，在第二碱试剂的存在下，式I化合物与式c所示的N-去甲基化试剂反应，得到式I-a化合物；
Figure PCTCN2021076337-appb-000019
其中，
环A为取代或未取代的4-12元碳环、取代或未取代的4-12元杂环、取代或未取代的5-12元杂芳环、取代或未取代的C 6-C 12芳环；
R 1和R 2各自独立为取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基；
X和Y各自独立地为碳原子、氧原子、硫原子或氮原子；
R 3为氢、卤素、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基，取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
R 12为取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的C 6-C 16芳基、取代或未取代的5-16元杂芳基、取代或未取代的C 1-C 6烷基-W-、取代或未取代的C 3-C 12环烷基-W-、取代或未取代的C 6-C 16芳基-W-、取代或未取代的5-16元杂芳基-W-；
R 13为卤素；
W为O或S；
n为1、2或3；
其中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、C 6-C 12芳基、5-10元杂芳基；
其中，所述的杂环、杂芳环和杂芳基各自独立地具有1-3个(优选为1、2或3个)选自N、O和S杂原子。
在另一优选例中，环A、X、Y、W、n、R 1、R 2、R 3、R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立地如本发明第一方面所述。
在另一优选例中，所述的第二溶剂中、在第二催化剂和式c所示的N-去甲基化试剂各自独立地如本发明第二方面所述。
在另一优选例中，所述的步骤(2)如本发明第二方面所述。
在另一优选例中，式I化合物、式c化合物和式I-a化合物各自独立地如本发明第二方面所述。
本发明第四方面，提供一种式I化合物的盐，
Figure PCTCN2021076337-appb-000020
其中，
环A为取代或未取代的5-7元碳环、取代或未取代的5-7元杂环、取代或未取代的5-7元杂芳环；
R 1和R 2各自独立为氢、取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基；
X和Y各自独立地为碳原子、氧原子、硫原子或氮原子；
W为O或S；
R 3为氢、卤素、取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基；
R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 6烷基、取 代或未取代的C 3-C 7环烷基；
n为1、2或3；
其中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、六元的芳基、五元或六元的杂芳基(优选C 5杂芳基)；
所述的杂环、杂芳环和杂芳基各自独立地具有1-3个(优选为1、2或3个)选自N、O和S杂原子。
在另一优选例中，环A、X、Y、W、n、R 1、R 2、R 3、R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立地如本发明第一方面所述。
在另一优选例中，所述的式I化合物具有式I-B所示的结构：
Figure PCTCN2021076337-appb-000021
在另一优选例中，所述的式I化合物具有式I-D所示的结构：
Figure PCTCN2021076337-appb-000022
在另一优选例中，所述的式I化合物选自下组：
Figure PCTCN2021076337-appb-000023
在另一优选例中，所述的式I化合物的盐为式I化合物与选自下组的酸形成的盐：盐酸、粘酸、D-葡萄糖醛酸、氢溴酸、氢氟酸、硫酸、硝酸、磷酸、甲酸、乙酸、丙酸、草酸、丙二酸、琥珀酸、富马酸、马来酸、乳酸、苹果酸、酒石酸、柠檬酸、苦味酸、甲磺酸、苯甲磺酸、苯磺酸、天冬氨酸、谷氨酸，或其组合。
在另一优选例中，所述的式I化合物的盐选自下组：盐酸盐、马来酸盐、草酸盐、粘酸盐、富马酸盐、D-葡萄糖醛酸盐，或其组合。
本发明第五方面，提供一种式Ic-1化合物的盐酸盐或盐酸盐晶型A，
Figure PCTCN2021076337-appb-000024
在另一优选例中，在Ic-1化合物的盐酸盐晶型A中，所述式Ic-1化合物与盐酸的分子摩尔比为4:1、3:1、2：1、1:1、1:2、1:3或4:1。
在另一优选例中，所述式Ic-1化合物的盐酸盐晶型A为无水晶型。
在另一优选例中，所述的盐酸盐晶型A的X-射线粉末衍射图谱在2θ角为18.27±0.2°、21.27±0.2°、22.89±0.2°处具有特征峰。
在另一优选例中，所述盐酸盐晶型A还在1个或多个选自下组的2θ值处具有特征峰：16.85±0.2°、22.20±0.2°、23.86±0.2°。
在另一优选例中，所述盐酸盐晶型A还在1个或多个选自下组的2θ值处具有特征峰：10.09±0.2°、25.40±0.2°、28.18±0.2°。
在另一优选例中，所述盐酸盐晶型A还在1个或多个选自下组的2θ值处具有特征峰：10.09±0.2°、16.85±0.2°、22.20±0.2°、23.86±0.2°、25.40±0.2°、28.18±0.2°。
在另一优选例中，所述盐酸盐晶型A还在1个或多个选自下组的2θ值处具有特征峰：11.25±0.2°、21.84±0.2°、26.76±0.2°、28.75±0.2°、32.57±0.2°。
在另一优选例中，所述盐酸盐晶型A在1个或多个选自下组的2θ值处具有特征峰： 10.09±0.2°、11.25±0.2°、16.85±0.2°、18.27±0.2°、21.27±0.2°、21.84±0.2°、22.20±0.2°、22.89±0.2°、23.86±0.2°、25.40±0.2°、26.76±0.2°、28.18±0.2°、28.75±0.2°、32.57±0.2°。
在另一优选例中，所述的盐酸盐晶型A的X-射线粉末衍射图谱具有一个或选自下组的2θ值处的特征峰和峰强度：
2θ值 d值 相对强度％
10.09 8.76 52.42
11.25 7.87 12.51
16.85 5.26 75.73
18.27 4.86 100.00
21.27 4.18 82.46
21.84 4.07 31.52
22.20 4.00 71.08
22.89 3.88 83.32
23.86 3.73 57.43
25.40 3.51 39.30
26.76 3.33 25.94
28.18 3.17 36.31
28.75 3.11 25.68
32.57 2.75 10.18。
在另一优选例中，所述盐酸盐晶型A具有基本如图8所示的X-射线粉末衍射特征峰。
在另一优选例中，所述盐酸盐晶型A的差示扫描量热分析(DSC)图在加热至141.8±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述盐酸盐晶型A的差示扫描量热分析(DSC)图基本如图9所示。
在另一优选例中，所述盐酸盐晶型A的热重分析(TGA)图在加热至120℃时具有约1.1±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述盐酸盐晶型A的热重分析(TGA)图基本如图9所示。
本发明第六方面，提供一种制备如本发明第五方面所述的式Ic-1化合物的盐酸盐晶型A的方法，所述的方法包括步骤：
(a)式Ic-1化合物与第一有机溶剂混合后，在5～15℃下滴加盐酸，调节体系pH为6-8，反应析出固体，过滤得到式Ic-1化合物的盐酸盐晶型A。
在另一优选中，所述的步骤(a)中，所述的第一有机溶剂包括乙酸乙酯。
在另一优选例中，所述的步骤(a)中，所述的盐酸为浓盐酸。
在另一优选例中，所述的步骤(a)中，体系pH为6.5-7.5，较佳地7.0。
在另一优选例中，所述的步骤(a)中，反应的时间为3-8min，较佳地5min。
在另一优选例中，所述的步骤(a)中，所述的反应在搅拌条件下反应。
在另一优选例中，所述的步骤(a)中，所述的盐酸缓慢的加入。
在另一优选例中，所述的步骤(a)中，式Ic-1化合物与第一有机溶剂的重量体积比(kg：L)为0.05-2:1-20，较佳地0.05-1:1-10，更佳地0.1-0.5：2-7。
在另一优选例中，所述的步骤(a)中，固体析出后，在40～45℃条件下干燥得到式Ic-1化合物的盐酸盐晶型A。
本发明第七方面，提供一种式Ic-1化合物的马来酸盐或马来酸盐晶型B，
Figure PCTCN2021076337-appb-000025
在另一优选例中，在Ic-1化合物的马来酸盐晶型B中，所述式Ic-1化合物与马来酸的分子摩尔比为4:1、3:1、2：1、1:1、1:2、1:3或4:1。
在另一优选例中，所述Ic-1化合物的马来酸盐晶型B为无水晶型。
在另一优选例中，所述的马来酸盐晶型B的X-射线粉末衍射图谱在2θ角为19.23±0.2°、24.04±0.2°、24.70±0.2°处具有特征峰。
在另一优选例中，所述马来酸盐晶型B还在1个或多个选自下组的2θ值处具有特征峰：11.83±0.2°、19.56±0.2°、28.15±0.2°。
在另一优选例中，所述马来酸盐晶型B还在1个或多个选自下组的2θ值处具有特征峰：9.70±0.2°、18.23±0.2°、24.93±0.2°。
在另一优选例中，所述马来酸盐晶型B还在1个或多个选自下组的2θ值处具有特征峰：9.70±0.2°、11.83±0.2°、18.23±0.2°、19.56±0.2°、24.93±0.2°、28.15±0.2°。
在另一优选例中，所述马来酸盐晶型B还在1个或多个选自下组的2θ值处具有特征峰：15.22±0.2°、16.30±0.2°、18.82±0.2°、21.15±0.2°、21.83±0.2°、23.60±0.2°、26.35±0.2°、28.94±0.2°、32.59±0.2°、33.31±0.2°、34.74±0.2°、35.94±0.2°、38.18。
在另一优选例中，所述马来酸盐晶型B在1个或多个选自下组的2θ值处具有特征峰：9.70±0.2°、11.83±0.2°、15.22±0.2°、16.30±0.2°、18.23±0.2°、18.82±0.2°、19.23±0.2°、19.56±0.2°、21.15±0.2°、21.83±0.2°、23.60±0.2°、24.04±0.2°、24.70±0.2°、24.93±0.2°、26.35±0.2°、28.15±0.2°、28.94±0.2°、32.59±0.2°、33.31±0.2°、34.74±0.2°、35.94±0.2°、38.18±0.2°。
在另一优选例中，所述的马来酸盐晶型B的X-射线粉末衍射图谱具有一个或选自下组的2θ值处的特征峰和峰强度：
2θ d值 相对强度％
9.70 9.12 15.51
11.83 7.48 18.79
15.22 5.82 10.21
16.30 5.44 13.25
18.23 4.87 14.69
18.82 4.72 11.41
19.23 4.61 72.02
19.56 4.54 37.68
21.15 4.20 8.57
21.83 4.07 1.84
23.60 3.77 12.70
24.04 3.70 100.00
24.70 3.60 42.79
24.93 3.57 15.64
26.35 3.38 12.47
28.15 3.17 24.14
28.94 3.09 9.61
32.59 2.75 3.74
33.31 2.69 3.29
34.74 2.58 2.77
35.94 2.50 1.13
38.18 2.36 1.62。
在另一优选例中，所述马来酸盐晶型B具有基本如图11所示的X-射线粉末衍射特征峰。
在另一优选例中，所述马来酸盐晶型B的差示扫描量热分析(DSC)图在加热至105.8±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述马来酸盐晶型B的差示扫描量热分析(DSC)图基本如图12所示。
在另一优选例中，所述马来酸盐晶型B的热重分析(TGA)图在加热至80℃时具有约0.8±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述马来酸盐晶型B的热重分析(TGA)图基本如图12所示。
本发明第八方面，提供一种制备如本发明第七方面所述的式Ic-1化合物的马来酸盐晶型B的方法，所述的方法包括步骤：
(b)式Ic-1化合物、马来酸和第二有机溶剂混合后，反应析出固体，过滤得到式Ic-1化合物的马来酸盐晶型B。
在另一优选中，所述的步骤(b)中，所述的第二有机溶剂包括乙酸乙酯。
在另一优选例中，所述的步骤(b)中，反应的时间为4-10天，较佳地5-7天。
在另一优选例中，所述的步骤(b)中，所述的反应在搅拌条件下反应。
在另一优选例中，所述搅拌的转速为700-800转/分钟。
在另一优选例中，所述的步骤(b)中，所述反应的温度为室温。
在另一优选例中，所述的步骤(b)中，式Ic-1化合物与第二有机溶剂的重量体积比(kg：ml)为0.05-2:1-10，较佳地0.05-1:1-30，更佳地0.1-0.5：1-20。
在另一优选例中，所述的步骤(b)中，式Ic-1化合物与马来酸的重量为0.05-2:0.05-0.3，较佳地0.05-1:0.05-0.2，更佳地0.1-0.5：0.08-0.16。
在另一优选例中，所述的步骤(b)中，固体析出后，室温真空干燥得到式Ic-1化合物的马来酸盐晶型B。
本发明第九方面，提供一种式Ic-1化合物的草酸盐或草酸盐晶型C，
Figure PCTCN2021076337-appb-000026
在另一优选例中，在Ic-1化合物的草酸盐晶型C中，所述式Ic-1化合物与草酸的分子摩尔比为4:1、3:1、2：1、1:1、1:2、1:3或4:1。
在另一优选例中，所述Ic-1化合物的草酸盐晶型C为无水晶型。
在另一优选例中，所述的草酸盐晶型C的X-射线粉末衍射图谱在2θ角为14.64±0.2°、22.05±0.2°、25.61±0.2°处具有特征峰。
在另一优选例中，所述草酸盐晶型C还在1个或多个选自下组的2θ值处具有特征峰：16.36±0.2°、20.90±0.2°、23.43±0.2°。
在另一优选例中，所述草酸盐晶型C还在1个或多个选自下组的2θ值处具有特征峰：15.27±0.2°、16.07±0.2°、19.52±0.2°。
在另一优选例中，所述草酸盐晶型C还在1个或多个选自下组的2θ值处具有特征峰：15.27±0.2°、16.07±0.2°、16.36±0.2°、17.63±0.2°、19.52±0.2°、20.90±0.2°、23.43±0.2°。
在另一优选例中，所述草酸盐晶型C在1个或多个选自下组的2θ值处具有特征峰：14.64±0.2°、15.27±0.2°、16.07±0.2°、16.36±0.2°、17.63±0.2°、19.52±0.2°、20.90±0.2°、22.05±0.2°、23.43±0.2°、25.61±0.2°。
在另一优选例中，所述的草酸盐晶型C的X-射线粉末衍射图谱具有一个或选自下组的2θ值处的特征峰和峰强度：
2θ d值 相对强度％
14.64 6.05 24.38
15.27 5.80 10.02
16.07 5.52 11.65
16.36 5.42 19.89
17.63 5.03 2.55
19.52 4.55 10.17
20.90 4.25 12.83
22.05 4.03 100.00
23.43 3.80 16.77
25.61 3.48 23.51。
在另一优选例中，所述草酸盐晶型C具有基本如图15所示的X-射线粉末衍射特征峰。
在另一优选例中，所述草酸盐晶型C的差示扫描量热分析(DSC)图在加热至152.2±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述草酸盐晶型C的差示扫描量热分析(DSC)图基本如图16所示。
在另一优选例中，所述草酸盐晶型C的热重分析(TGA)图在加热至100℃时具有约1.0±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述草酸盐晶型C的热重分析(TGA)图基本如图16所示。
本发明第十方面，提供一种制备本发明第九方面所述的式Ic-1化合物的草酸盐晶型C的方法，所述的方法包括步骤：
(c)式Ic-1化合物、草酸和第三有机溶剂混合后，反应析出固体，过滤得到式Ic-1化合物的草酸盐晶型C。
在另一优选中，所述的步骤(c)中，所述的第三有机溶剂包括乙酸乙酯。
在另一优选例中，所述的步骤(c)中，反应的时间为4-10天，较佳地5-7天。
在另一优选例中，所述的步骤(c)中，所述的反应在搅拌条件下反应。
在另一优选例中，所述搅拌的转速为700-800转/分钟。
在另一优选例中，所述的步骤(c)中，所述反应的温度为室温。
在另一优选例中，所述的步骤(c)中，式Ic-1化合物与第三有机溶剂的重量体积比(kg：ml)为0.05-2:1-10，较佳地0.05-1:1-30，更佳地0.1-0.5：1-20。
在另一优选例中，所述的步骤(c)中，式Ic-1化合物与草酸的重量为0.05-2:0.05-0.3，较佳地0.08-1:0.05-0.2，更佳地0.1-0.5：0.06-0.13。
在另一优选例中，所述的步骤(c)中，固体析出后，室温真空干燥得到式Ic-1 化合物的草酸盐晶型C。
本发明第十一方面，提供一种式Ic-1化合物的粘酸盐或粘酸盐晶型D，
Figure PCTCN2021076337-appb-000027
在另一优选例中，在Ic-1化合物的粘酸盐晶型D中，所述式Ic-1化合物与粘酸的分子摩尔比为4:1、3:1、2：1、1:1、1:2、1:3或4:1。
在另一优选例中，所述Ic-1化合物的粘酸盐晶型D为无水晶型。
在另一优选例中，所述的粘酸盐晶型D的X-射线粉末衍射图谱在2θ角为3.79±0.2°、11.28±0.2°、19.48±0.2°处具有特征峰。
在另一优选例中，所述粘酸盐晶型D还在1个或多个选自下组的2θ值处具有特征峰：15.81±0.2°、20.98±0.2°、23.91±0.2°。
在另一优选例中，所述粘酸盐晶型D还在1个或多个选自下组的2θ值处具有特征峰：16.97±0.2°、25.88±0.2°、28.40±0.2°。
在另一优选例中，所述粘酸盐晶型D还在1个或多个选自下组的2θ值处具有特征峰：15.81±0.2°、16.97±0.2°、20.98±0.2°、23.91±0.2°、25.88±0.2°、28.40±0.2°。
在另一优选例中，所述粘酸盐晶型D还在1个或多个选自下组的2θ值处具有特征峰：14.21±0.2°、17.71±0.2°、27.16±0.2°、29.49±0.2°、30.74±0.2°、32.33±0.2°、34.50±0.2°、35.42±0.2°、36.16±0.2°。
在另一优选例中，所述粘酸盐晶型D在1个或多个选自下组的2θ值处具有特征峰：3.79±0.2°、11.28±0.2°、14.21±0.2°、15.81±0.2°、16.97±0.2°、17.71±0.2°、19.48±0.2°、20.98±0.2°、23.91±0.2°、25.88±0.2°、27.16±0.2°、28.40±0.2°、29.49±0.2°、30.74±0.2°、32.33±0.2°、34.50±0.2°、35.42±0.2°、36.16±0.2°。
在另一优选例中，所述的粘酸盐晶型D的X-射线粉末衍射图谱具有一个或选自下组的2θ值处的特征峰和峰强度：
2θ d值 相对强度％
3.79 23.30 40.31
11.28 7.84 100.00
14.21 6.23 3.82
15.81 5.61 39.14
16.97 5.23 14.93
17.71 5.01 8.72
19.48 4.56 87.36
20.98 4.23 18.69
23.91 3.72 21.51
25.88 3.44 10.60
27.16 3.28 3.35
28.40 3.14 10.06
29.49 3.03 5.70
30.74 2.91 6.11
32.33 2.77 3.19
34.50 2.60 3.87
35.42 2.53 3.41
36.16 2.48 2.23。
在另一优选例中，所述粘酸盐晶型D具有基本如图18所示的X-射线粉末衍射特征峰。
在另一优选例中，所述粘酸盐晶型D的差示扫描量热分析(DSC)图在加热至140.9±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述粘酸盐晶型D的差示扫描量热分析(DSC)图基本如图19所示。
在另一优选例中，所述粘酸盐晶型D的热重分析(TGA)图在加热至100℃时具有约1.45±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述粘酸盐晶型D的热重分析(TGA)图基本如图19所示。
本发明第十二方面，提供一种制备本发明第十一方面所述的式Ic-1化合物的粘酸盐晶型D的方法，所述的方法包括步骤：
(d)式Ic-1化合物、粘酸和第四有机溶剂混合后，反应析出固体，过滤得到式Ic-1化合物的粘酸盐晶型D。
在另一优选中，所述的步骤(d)中，所述的第四有机溶剂包括乙酸乙酯。
在另一优选中，所述的步骤(d)中，所述反应的温度为40-60℃。
在另一优选中，所述的步骤(d)中，所述反应的时间为1-3天，较佳地1.5-2.5天。
在另一优选例中，所述的步骤(d)中，粘酸与式Ic-1化合物的重量体积比(mg:ml)为1-20:0.1-1.5，较佳地1-15:0.1-1，更佳地5-10:0.2-0.8。
在另一优选例中，所述的步骤(d)中，在混合后的体系中，式Ic-1化合物在第四溶剂中的浓度为20-60mg/mL，较佳地30-50mg/mL，更佳地35-45mg/mL。
在另一优选例中，所述的步骤(d)中，固体析出后，室温真空干燥得到式Ic-1化合物的粘酸晶型D。
本发明第十三方面，提供一种式Ic-1化合物的富马酸盐或富马酸盐晶型E，
Figure PCTCN2021076337-appb-000028
在另一优选例中，在Ic-1化合物的富马酸盐晶型E中，所述式Ic-1化合物与富马酸的分子摩尔比为4:1、3:1、2：1、1:1、1:2、1:3或4:1。
在另一优选例中，所述Ic-1化合物的富马酸盐晶型E为无水晶型。
在另一优选例中，所述的富马酸盐晶型E的X-射线粉末衍射图谱在2θ角为14.17±0.2°、18.95±0.2°、23.76±0.2°处具有特征峰。
在另一优选例中、所述富马酸盐晶型E还在1个或多个选自下组的2θ值处具有特征峰：4.7±0.2°、22.75±0.2°、26.93±0.2°。
在另一优选例中，所述富马酸盐晶型E还在1个或多个选自下组的2θ值处具有特征峰：13.34±0.2°、15.63±0.2°、28.69±0.2°。
在另一优选例中，所述富马酸盐晶型E还在1个或多个选自下组的2θ值处具有特征峰：4.7±0.2°、13.34±0.2°、15.63±0.2°、22.75±0.2°、26.93±0.2°、28.69±0.2°。
在另一优选例中，所述富马酸盐晶型E还在1个或多个选自下组的2θ值处具有特征峰：9.41±0.2°、17.62±0.2°、25.66±0.2°、31.34±0.2°。
在另一优选例中，所述富马酸盐晶型E在1个或多个选自下组的2θ值处具有特征峰：4.7±0.2°、9.41±0.2°、13.34±0.2°、14.17±0.2°、15.63±0.2°、17.62,18.95±0.2°、22.75±0.2°、23.76±0.2°、25.66±0.2°、26.93±0.2°、28.69±0.2°、31.34±0.2°。
在另一优选例中，所述的富马酸盐晶型E的X-射线粉末衍射图谱具有一个或选自下组的2θ值处的特征峰和峰强度：
2θ d值 相对强度％
4.70 18.82 9.14
9.41 9.40 5.61
13.34 6.64 7.80
14.17 6.25 28.48
15.63 5.67 6.40
17.62 5.03 2.92
18.95 4.68 44.00
22.75 3.91 10.62
23.76 3.74 100.00
25.66 3.47 5.88
26.93 3.31 14.03
28.69 3.11 6.48
31.34 2.85 2.76。
在另一优选例中，所述富马酸盐晶型E具有基本如图21所示的X-射线粉末衍射特征峰。
在另一优选例中，所述富马酸盐晶型E的差示扫描量热分析(DSC)图在加热至76.5±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述富马酸盐晶型E的差示扫描量热分析(DSC)图基本如图22所示。
在另一优选例中，所述富马酸盐晶型E的热重分析(TGA)图在加热至80℃时具有约2.17±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述富马酸盐晶型E的热重分析(TGA)图基本如图22所示。
本发明第十四方面，提供一种制备本发明第十三方面所述的式Ic-1化合物的富马酸盐晶型E的方法，所述的方法包括步骤：
(e)式Ic-1化合物、富马酸和第五有机溶剂混合后，反应析出固体，过滤得到式Ic-1化合物的富马酸盐晶型E。
在另一优选中，所述的步骤(e)中，所述的第五有机溶剂包括乙酸乙酯。
在另一优选中，所述的步骤(e)中，混合后室温搅拌3-5天。
在另一优选例中，所述的步骤(e)中，所述反应的温度为室温，
在另一优选例中，所述的步骤(e)中，富马酸与式Ic-1化合物的重量体积比(mg:ml)为1-20:0.1-1.5，较佳地1-15:0.1-1，更佳地5-10:0.2-0.8。
在另一优选例中，所述的步骤(e)中，在混合后的体系中，式Ic-1化合物在第五溶剂中的浓度为20-60mg/mL，较佳地30-50mg/mL，更佳地35-45mg/mL。
在另一优选例中，所述的步骤(e)中，固体析出后，室温真空干燥得到式Ic-1化合物的富马酸晶型E。
本发明第十五方面，提供一种式Ic-1化合物的D-葡萄糖醛酸盐或D-葡萄糖醛酸盐晶型F，
Figure PCTCN2021076337-appb-000029
在另一优选例中，在D-葡萄糖醛酸盐晶型F中，所述式Ic-1化合物与D-葡萄糖醛酸的分子摩尔比为4:1、3:1、2：1、1:1、1:2、1:3或4:1。
在另一优选例中，所述Ic-1化合物的D-葡萄糖醛酸盐晶型F为无水晶型。
在另一优选例中，所述的D-葡萄糖醛酸盐晶型F的X-射线粉末衍射图谱在2θ角为4.77±0.2°、16.13±0.2°、19.53±0.2°处具有特征峰。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F还在1个或多个选自下组的2θ值处具有特征峰8.34±0.2°、17.54±0.2°、20.06±0.2°。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F还在1个或多个选自下组的2θ值处具有特征峰：10.87±0.2°、21.25±0.2°、25.93±0.2°。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F还在1个或多个选自下组的2θ值处具有特征峰：8.34±0.2°、10.87±0.2°、17.54±0.2°、20.06±0.2°、21.25±0.2°、23.42±0.2°、25.93±0.2°。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F在1个或多个选自下组的2θ值处具有特征峰：4.77±0.2°、8.34±0.2°、10.87±0.2°、16.13±0.2°、17.54±0.2°、19.53±0.2°、20.06±0.2°、21.25±0.2°、23.42±0.2°、25.93±0.2°。
在另一优选例中，所述的D-葡萄糖醛酸盐晶型F的X-射线粉末衍射图谱具有一个或选自下组的2θ值处的特征峰和峰强度：
2θ d值 相对强度％
4.77 18.52 100.00
8.34 10.60 54.57
10.87 8.14 46.79
16.13 5.49 97.98
17.54 5.06 55.33
19.53 4.55 74.17
20.06 4.43 60.75
21.25 4.18 44.80
23.42 3.80 29.68
25.93 3.44 31.65。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F具有基本如图24所示的X-射线粉末衍射特征峰。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的差示扫描量热分析(DSC)图在加热至119.1±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的差示扫描量热分析(DSC)图基本如图25所示。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的热重分析(TGA)图在加热至 100℃时具有约2.71±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的热重分析(TGA)图基本如图25所示。
本发明第十六方面，一种制备本发明第十五方面所述的式Ic-1化合物的D-葡萄糖醛酸盐晶型F的方法，所述的方法包括步骤：
(f)式Ic-1化合物、D-葡萄糖醛酸和第六有机溶剂混合后，反应析出固体，过滤得到式Ic-1化合物的D-葡萄糖醛酸盐晶型F。
在另一优选中，所述的步骤(f)中，所述的第六有机溶剂包括乙腈。
在另一优选中，所述的步骤(f)中，混合后室温搅拌3-5天。
在另一优选例中，所述的步骤(f)中，所述反应的温度为室温，
在另一优选例中，所述的步骤(f)中，D-葡萄糖醛酸与式Ic-1化合物的重量体积比(mg:ml)为5-30:0.1-2，较佳地5-20:0.1-1，更佳地10-16:0.2-0.7。
在另一优选例中，所述的步骤(f)中，在混合后的体系中，式Ic-1化合物在第五溶剂中的浓度为20-60mg/mL，较佳地30-50mg/mL，更佳地35-45mg/mL。
在另一优选例中，所述的步骤(f)中，固体析出后，室温真空干燥得到式Ic-1化合物的D-葡萄糖醛酸晶型F。
本发明第十七方面，提供一种药物组合物，所述的组合物包括如本发明第五方面所述的式Ic-1化合物的盐酸盐晶型A、本发明第七方面所述的式Ic-1化合物的马来酸盐晶型B、本发明第九方面所述的式Ic-1化合物的草酸盐晶型C、本发明第十一方面所述的式Ic-1化合物的粘酸盐晶型D、本发明第十三方面所述的式Ic-1化合物的富马酸盐晶型E、或本发明第十五方面所述的式Ic-1化合物的D-葡萄糖醛酸盐晶型F；和药学上可接受的载体。
本发明第十八方面，提供一种如本发明第五方面所述的式Ic-1化合物的盐酸盐晶型A、本发明第七方面所述的式Ic-1化合物的马来酸盐晶型B、本发明第九方面所述的式Ic-1化合物的草酸盐晶型C、本发明第十一方面所述的式Ic-1化合物的粘酸盐晶型D、本发明第十三方面所述的式Ic-1化合物的富马酸盐晶型E、或本发明第十五方面所述的式Ic-1化合物的D-葡萄糖醛酸盐晶型F的用途，用于(a)制备瞬时受体电位通道蛋白(TRP)抑制剂；和/或(b)制备预防和/或治疗与瞬时受体电位通道蛋白(TRP)相关的疾病的药物。
在另一优选例中，所述的瞬时受体电位通道蛋白(TRP)为TRPA1。
在另一优选例中，与瞬时受体电位通道蛋白(TRP)相关的疾病选自下组：疼痛、癫痫、炎症、呼吸障碍、瘙痒、尿路障碍、炎症性肠病，或其组合。
在另一优选例中，所述的疼痛为急性疼痛或慢性疼痛。
在另一优选例中，所述的疼痛选自下组：急性炎性疼痛、慢性炎性疼痛、内脏痛、神经源性疼痛、肌纤维痛、头痛、神经痛、癌症引起疼痛，或其组合。
在另一优选例中，所述的疼痛为炎症疼痛。
在另一优选例中，所述的炎性疼痛急性炎性疼痛或慢性炎性疼痛。
在另一优选例中，所述的头痛为偏头痛或肌紧张性疼痛。
在另一优选例中，所述的神经痛为三叉神经痛、糖尿病性疼痛或带状孢疹后神经痛。
在另一优选例中，所述的疼痛选自下组：急性疼痛、肌纤维痛、内脏疼痛、炎症疼痛、神经痛，或其组合。
在另一优选例中，所述的疼痛为肌纤维痛。
本发明第十九方面，提供一种体外非治疗性和非诊断性的抑制瞬时受体电位通道蛋白活性的方法，包括步骤：将瞬时受体电位通道蛋白或表达所述蛋白的细胞与如本发明第五方面所述的式Ic-1化合物的盐酸盐晶型A、本发明第七方面所述的式Ic-1化合物的马来酸盐晶型B、本发明第九方面所述的式Ic-1化合物的草酸盐晶型C、本发明第十一方面所述的式Ic-1化合物的粘酸盐晶型D、本发明第十三方面所述的式Ic-1化合物的富马酸盐晶型E、或本发明第十五方面所述的式Ic-1化合物的D-葡萄糖醛酸盐晶型F进行接触，从而抑制瞬时受体电位通道蛋白的活性。
本发明第二十方面，提供一种抑制瞬时受体电位通道蛋白或预防和/或治疗与瞬时受体电位通道蛋白(TRP)相关的疾病的方法，包括步骤：给需要的对象施用如如本发明第五方面所述的式Ic-1化合物的盐酸盐晶型A、本发明第七方面所述的式Ic-1化合物的马来酸盐晶型B、本发明第九方面所述的式Ic-1化合物的草酸盐晶型C、本发明第十一方面所述的式Ic-1化合物的粘酸盐晶型D、本发明第十三方面所述的式Ic-1化合物的富马酸盐晶型E、或本发明第十五方面所述的式Ic-1化合物的D-葡萄糖醛酸盐晶型F，从而预防和/或治疗与瞬时受体电位通道蛋白(TRP)相关的疾病。
在另一优选例中，所述对象包括人和非人哺乳动物(啮齿动物、兔、猴、家畜、狗、猫等)。
应理解，在本发明范围内，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1A-1E为本发明化合物I C-3、I C-4、I C-8I C-23和I C-24抑制TRPA1活性的量效关系曲线图。
图2为本发明化合物I C-10在小鼠福尔马林疼痛模型中的镇痛活性结果。
图3为本发明化合物I C-23在小鼠热板致痛模型中的镇痛活性结果。
图4为化合物Ic-1和度洛西汀在小鼠肌纤维痛模型中的镇痛活性结果，Mean±SD,n＝12,**p<0.01，***p<0.001与溶剂对照组比较。
图5为化合物I C-1、度洛西汀、吲哚美辛和山莨胆碱在小鼠醋酸扭体痛模型中的镇痛活性结果。
图6为化合物IC-1和加巴喷丁在大鼠SNL模型中的镇痛活性结果。
图7为化合物Ic-1和度洛西汀在小鼠福尔马林模型中不同给药剂量下在II相(10-60min)期添足时间统计结果。
图8为式Ic-1化合物的盐酸盐晶型A的XRPD图。
图9为式Ic-1化合物的盐酸盐晶型A的TGA/DSC图。
图10为式Ic-1化合物的盐酸盐晶型A的变温XRPD对比图。
图11为式Ic-1化合物的马来酸盐晶型B的XRPD图。
图12为式Ic-1化合物的马来酸盐晶型B的TGA/DSC图。
图13为式Ic-1化合物的马来酸盐晶型B的1H NMR图谱。
图14为式Ic-1化合物的马来酸盐晶型B的变温XRPD对比图。
图15为式Ic-1化合物的草酸盐晶型C的XRPD图。
图16为式Ic-1化合物的草酸盐晶型C的TGA/DSC图。
图17为式Ic-1化合物的草酸盐晶型C的变温XRPD对比图。
图18为式Ic-1化合物的粘酸盐晶型D的XPRD图。
图19为式Ic-1化合物的粘酸盐晶型D的TGA/DSC图。
图20为式Ic-1化合物的粘酸盐晶型D的1H NMR图谱。
图21为式Ic-1化合物的富马酸盐晶型E的XRPD图。
图22为式Ic-1化合物的富马酸盐晶型E的TGA/DSC图。
图23为式Ic-1化合物的富马酸盐晶型E的1H NMR图谱。
图24为式Ic-1化合物的D-葡萄糖醛酸盐晶型F的XRPD图。
图25为式Ic-1化合物的D-葡萄糖醛酸盐晶型F的TGA/DSC图。
图26为式Ic-1化合物的盐酸盐晶型A的DVS图。
图27为式Ic-1化合物的盐酸盐晶型A DVS测试前后的XRPD对比图。
图28为式Ic-1化合物的马来酸盐晶型B的DVS图。
图29为式Ic-1化合物的马来酸盐晶型B测试DVS前后的XRPD对比图。
图30为式Ic-1化合物的盐酸盐晶型A稳定性测试前后的XRPD对比图。
图31为式Ic-1化合物的马来酸盐晶型B稳定性测试前后的XRPD对比图。
图32为式Ic-1化合物的盐酸盐晶型A的单晶衍射图。
具体实施方式
本发明人通过广泛而深入的研究，首次意外地开发了一种化合物，或其药学上可接受的盐及其制备方法和盐晶型，所述化合物具有式I结构。实验表明，本发明化合物对TRP通道具有显著的抑制效果。本发明的化合物可有效治疗与TRP(尤其是TRPA1)靶点相关的疼痛等。在此基础上，完成了本发明。
术语
如本文所用，术语“包含”、“包括”、“含有”可互换使用，不仅包括封闭式定义，还包括半封闭、和开放式的定义。换言之，所述术语包括了“由……构成”、“基本上由……构成”。
如本文所用，“R 1”、“R 1”和“R1”的含义相同，可相互替换，其它类似定义的含义相同。
如本文所用，术语“C 1-C 12烷基”、“C 1-C 8烷基”、“C 1-C 6烷基”、“C 1-C 4烷基”、或“C 1-C 3烷基”指具有1-12、1-8、1-6、1-4或1-3个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基，或类似基团。
如本文所用，术语“C 1-C 6烷氧基”指具有1-6碳原子的直链或支链的烷氧基，例如甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、仲丁氧基、叔丁氧基、戊氧基、己氧基，或类似基团。
如本文所用，术语“C 6-C 12苯并脂肪环基”指具有6-12个碳原子的基团，包括茚满基、四氢萘基或二氢萘基等类似基团。
如本文所用，术语“C 3-C 12环烷基”或“C 3-C 7环烷基”指具有3-12个或3-7个碳原子的环烷基(包括单环、二环或多环环系)，例如环丙基、环丁基、甲基环丁基、环戊基、环庚基，或类似基团。
如本文所用，术语“C 2-C 6酯基”指具有C 1-C 5烷基-COO-结构的基团或者具有-COO-C 1～C 5烷基结构的基团，其中烷基可以为直链或支链的，例如CH 3COO-、C 2H 5COO-、C 3H 8COO-、(CH 3)2CHCOO-、-COOCH 3、-COOC 2H 5、-COOC 3H 8，或类似基团。
如本文所用，术语“C 2-C 4酰胺基”指具有C 1-C 3烷基-CO-NH-结构的基团或者具有-CO-NH-C 1-C 3烷基结构的基团，其中烷基可以为直链或支链的，例如CH 3-CO-NH-、C 2H 5-CO-NH-、C 3H 8-CO-NH-、-COOCH 3、-CO-NH-C 2H 5、-CO-NH-C 3H 8，或类似基 团。
如本文所用，术语“C 2-C 4酰基”指具有C 1-C 3烷基-CO-结构的基团其中烷基可以为直链或支链的，例如CH 3-CO-、C 2H 5-CO-、C 3H 8-CO-，或类似基团。
如本文所用，术语“C 3-C 7杂环烷基”指具有3-7个环碳原子和1-3个杂原子(优选含有1个氮原子，即与R 1和R 2共同相邻的氮原子)的单环和多环杂环(优选单环杂环)，例如哌啶基团、四氢吡咯基，或相似基团。
如本文所用，术语“4-12元碳环”、“5-10元碳环”或“5-7元碳环”为任何稳定的4、5、6、7、8、9、10、11或12元单环、二环或多环，碳环可以是饱和的、部分不饱和的、不饱和的环，但不能为芳族的环。所述碳环的实例包括但不限于环丙环、环丁环、环丁烯环、环戊环、环戊烯环、环己环、环己烯环、环庚环、环庚烯环、金刚烷环、环辛环、环辛烯环、环辛二烯环、二环[3.3.0]辛烷、二环[4.3.0]壬烷、二环[4.4.0]癸烷、二环[2.2.2]辛烷、芴基、茚满环。
如本文所用，术语“杂环”为任何稳定的单环、二环或多环(例如5、6或7元)，杂环上含有一个或多个(如1-3)选自N、O和S的杂原子，杂环可以是饱和的、部分不饱和的、不饱和的环，但不能为芳族的环。杂环前面的元数是指环原子数，例如4-12元杂环具有4-12个环原子。
如本文所用，术语“杂芳环”指具有一个到多个(优选为1、2或3个)选自N、O和S杂原子的芳族杂环系。杂芳环前面的元数是指环原子数，例如5-7元杂芳环具有5-7个环原子。应当理解的是，当含有多个杂原子时，杂原子可以相同、可以部分相同，也可以完全不同。例如5元杂芳环的实例包括(但不限于)：吡咯环、呋喃环、噻吩环、咪唑环、恶唑环、噻唑环，6元杂芳环的实例包括(但不限于)吡啶环、吡嗪环、哒嗪环、嘧啶环。或相似基团。
如本文所用，术语“C 1-C 6卤代烷基”和“C 1-C 3卤代烷基”是指具有1-6和1-3个碳原子的直链或支链的烷基的一个或多个氢原子被卤素基团取代，如一氯甲烷、二氯乙烷、三氯丙烷，或类似基团。
如本文所用，术语“C 1-C 6卤代烷氧基”是指具有1-6个碳原子的直链或支链的烷氧基的一个或多个氢原子被卤素基团取代，如一氯甲氧基、二氯氧基，或类似基团。
如本文所用，术语“C 1-C 4·羧基”是指C 1-C 3烷基-COOH结构的基团，其中烷基可以为直链或支链的，例如CH 3COOH、C 2H 5COOH、C 3H 8COOH、(CH 3) 2CHCOOH，或类似基团。
如本文所用，术语“C 6-C 12芳基”是指在环部分中具有6至12个碳原子的单环或二环芳族烃基，诸如苯基、萘基、联苯基，或相似基团。
如本文所用，术语“芳环”指芳族环系。芳环前面的碳数是指环原子数，例如“C 6-C 12芳环”指在环部分中具有6至12个碳原子的单环或二环芳族环，代表性地，C 6-C 12芳环为苯环或萘环。
如本文所用，术语“杂芳基”是指任选取代的芳香基，例如其为5至7元单环环系，所述环系具有的环含有至少一个杂原子和至少一个碳原子。杂芳基前面的元数是指环原子数，例如5-12元杂芳基是指具有5-12个环原子的杂芳基，包括(但不限于)：吡咯基、噻吩基、吡啶基、哒嗪基、嘧啶基、吡嗪基、呋喃基、咪唑基、噻唑基、噁唑基、三氮唑基，或类似基团。
如本文所用，术语“卤素”指F、Cl、Br和I。
如本文所用，术语“取代”是基团上的氢原子被非氢原子基团取代，但需要满足其化合价要求并且由取代生成化学稳定的化合物。在本说明书中，应解释为所有取代基为未取代的，除非在本文中明确描述为“取代的”。
如本文所用，
Figure PCTCN2021076337-appb-000030
与
Figure PCTCN2021076337-appb-000031
含义相同，均表示具有未取代的或具有1-5个(优选1-3个)R 3取代基的杂芳基。
如本文所用，
Figure PCTCN2021076337-appb-000032
为连接位点。
同样，应当理解的是，在本发明中，取代基可以在任何原子上与母体基团或底物连接，除非其连接违反化合价要求；母体基团或底物的氢原子可以在同一原子上，也可以在不同的原子上。
如本文所用，室温指的是25±5℃。
如本文所用，对于一个数值范围P1至P2,则该范围不仅包括端点P1和P2，还包括介于端点P1和P2的任何数值点。此外，对于P1和P2均为正数时，则对于一个整数n，该数值范围包括介于端点P1和P2的任何整数数值点。例如，对于一个整数n，当其数值范围为1-10时，包括1、2、3、4、5、6、7、8、9、和10；数值范围3-7包括3、4、5、6、7。代表性地，对于基团而言，C3-C7包括C3、C4、C5C6、和C7。
活性成分
如本文所用，“本发明化合物”、“本发明的3-芳氧基-3-芳香基-丙胺类化合物”、或“式I化合物”可互换使用，指具有式I结构的化合物，或其药学上可接受的盐。应理解，该术语还包括上述组分的混合物；
Figure PCTCN2021076337-appb-000033
其中：
环A为取代或未取代的4-12元碳环、取代或未取代的4-12元杂环、取代或未取代的5-12元杂芳环、取代或未取代的C 6-C 12芳环；
R 1和R 2各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
X和Y各自独立地为碳原子、氧原子、硫原子或氮原子；
R 3为氢、卤素、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基，取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
W为O或S；
n为1、2或3；
其中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、C 6-C 12芳基、5-10元杂芳基。
其中，所述的杂环、杂芳环和杂芳基各自独立地具有1-3个(优选为1、2或3个)选自N、O和S杂原子。
在另一优选例中，环A、X、Y、W、n、R 1、R 2、R 3、R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立地如上述本发明第一方面所述。
本发明化合物不仅对TRPA1具有抑制作用，对TRP家族中其它成员也有一定的抑制作用。
术语“药学上可接受的盐”指本发明化合物与酸或碱所形成的适合用作药物的盐。药学上可接受的盐包括无机盐和有机盐。一类优选的盐是本发明化合物与酸形成的盐，适合形成盐的酸包括(但并不限于)：盐酸、氢溴酸、氢氟酸、硫酸、硝酸、磷酸等无机酸，甲酸、乙酸、丙酸、草酸、丙二酸、琥珀酸、富马酸、马来酸、乳酸、苹果酸、酒石酸、柠檬酸、苦味酸、甲磺酸、苯甲磺酸，苯磺酸等有机酸；以及天冬氨酸、谷氨酸等酸性氨基酸。一类优选的盐是本发明化合物与碱形成的金属盐，适合形成盐的碱包括(但并不限于)：氢氧化钠、氢氧化钾、碳酸钠、碳酸氢钠、磷酸钠等无机碱、氨水、三乙胺、二乙胺等有机碱。
本发明所述的如式I所示化合物可通过常规方法转化为其药学上可接受的盐，例如，可将相应的酸的溶液加入到上述化合物的溶液中，成盐完全后减压除去溶剂即得本发明所述化合物的相应的盐。
在另一优选例中，所述的式I化合物如下表1所示：
表1
Figure PCTCN2021076337-appb-000034
制备方法
本发明提供一种式I结构的化合物，或其药学上可接受的盐的制备方法，所述的方法包括步骤：
(1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式a化合物与式b化合物反应，得到式I化合物；`
Figure PCTCN2021076337-appb-000035
其中，所述步骤(1)中各个反应参数(如第一溶剂中，在第一催化剂和第一碱试剂)如上述本发明第一方面所述。
优选地，本发明提供式Ic-1化合物的制备方法，所述的方法包括步骤：
(1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式i化合物与式ii化合物反应，得到式Ic-8化合物；
Figure PCTCN2021076337-appb-000036
(2)在第二溶剂中，在第二碱试剂的存在下，式Ic-化合物与氯甲酸苯酯反应，得到式iii化合物；
Figure PCTCN2021076337-appb-000037
(3)在第三溶剂中，在第三碱试剂的存在下，式iii化合物发生水解反应，得到式Ic-1化合物；
Figure PCTCN2021076337-appb-000038
其中，所述步骤(1)中各个反应参数(如第一溶剂中，在第一催化剂和第一碱试剂)如上述本发明第二方面所述。
晶型
本发明提供一种式Ic-1化合物的盐晶型。
如本文所用，术语“式Ic-1化合物的盐酸盐晶型A”、“盐酸盐晶型A”与“晶型A”可互换使用。
如本文所用，术语“式Ic-1化合物的马来酸盐晶型B”、“马来酸盐晶型B”与“晶型B”可互换使用。
如本文所用，术语“式Ic-1化合物的草酸盐晶型C”、“草酸盐晶型C”与“晶型C”可互换使用。
如本文所用，术语“式Ic-1化合物的粘酸盐晶型D”、“粘酸盐晶型D”与“晶型D”可互换使用。
如本文所用，术语“式Ic-1化合物的富马酸盐晶型E”、“富马酸盐晶型E”与“晶型E”可互换使用。
如本文所用，术语“式Ic-1化合物的D-葡萄糖醛酸盐晶型F”、“D-葡萄糖醛酸盐晶型F”与“晶型F”可互换使用。
代表性地，本发明提供一种式Ic-1化合物的盐酸盐晶型A，所述盐酸盐晶型A在1个或多个选自下组的2θ值处具有特征峰：10.09±0.2°、11.25±0.2°、16.85±0.2°、18.27±0.2°、21.27±0.2°、21.84±0.2°、22.20±0.2°、22.89±0.2°、23.86±0.2°、25.40±0.2°、26.76±0.2°、28.18±0.2°、28.75±0.2°、32.57±0.2°。
在另一优选例中，所述盐酸盐晶型A具有基本如图8所示的X-射线粉末衍射特征峰。
在另一优选例中，所述盐酸盐晶型A的差示扫描量热分析(DSC)图在加热至141.8±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述盐酸盐晶型A的差示扫描量热分析(DSC)图基本如图9所示。
在另一优选例中，所述盐酸盐晶型A的热重分析(TGA)图在加热至120℃时具有约1.1±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述盐酸盐晶型A的热重分析(TGA)图基本如图9所示。
代表性地，本发明提供一种式Ic-1化合物的马来酸盐晶型B，所述马来酸盐晶型B在1个或多个选自下组的2θ值处具有特征峰：9.70±0.2°、11.83±0.2°、15.22±0.2°、16.30±0.2°、18.23±0.2°、18.82±0.2°、19.23±0.2°、19.56±0.2°、21.15±0.2°、21.83±0.2°、23.60±0.2°、24.04±0.2°、24.70±0.2°、24.93±0.2°、26.35±0.2°、28.15±0.2°、28.94±0.2°、32.59±0.2°、33.31±0.2°、34.74±0.2°、35.94±0.2°、38.18±0.2°。
马来酸的结构如下：
Figure PCTCN2021076337-appb-000039
在另一优选例中，所述马来酸盐晶型B具有基本如图11所示的X-射线粉末衍射特征峰。
在另一优选例中，所述马来酸盐晶型B的差示扫描量热分析(DSC)图在加热至105.8±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述马来酸盐晶型B的差示扫描量热分析(DSC)图基本如图12所示。
在另一优选例中，所述马来酸盐晶型B的热重分析(TGA)图在加热至80℃时具有约0.8±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述马来酸盐晶型B的热重分析(TGA)图基本如图12所示。
代表性地，本发明提供一种式Ic-1化合物的草酸盐晶型C，所述草酸盐晶型C在1个或多个选自下组的2θ值处具有特征峰：14.64±0.2°、15.27±0.2°、16.07±0.2°、16.36±0.2°、17.63±0.2°、19.52±0.2°、20.90±0.2°、22.05±0.2°、23.43±0.2°、25.61±0.2°。
草酸又称为乙二酸，结构如下：
Figure PCTCN2021076337-appb-000040
在另一优选例中，所述草酸盐晶型C具有基本如图15所示的X-射线粉末衍射特征峰。
在另一优选例中，所述草酸盐晶型C的差示扫描量热分析(DSC)图在加热至152.2±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述草酸盐晶型C的差示扫描量热分析(DSC)图基本如图16所示。
在另一优选例中，所述草酸盐晶型C的热重分析(TGA)图在加热至100℃时具有约1.0±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述草酸盐晶型C的热重分析(TGA)图基本如图16所示。
代表性地，本发明提供一种式Ic-1化合物的粘酸盐晶型D，在另一优选例中，所述粘酸盐晶型D在1个或多个选自下组的2θ值处具有特征峰：3.79±0.2°、11.28±0.2°、14.21±0.2°、15.81±0.2°、16.97±0.2°、17.71±0.2°、19.48±0.2°、20.98±0.2°、23.91±0.2°、25.88±0.2°、27.16±0.2°、28.40±0.2°、29.49±0.2°、30.74±0.2°、32.33±0.2°、34.50±0.2°、35.42±0.2°、36.16±0.2°。
粘酸的结构式如下
Figure PCTCN2021076337-appb-000041
在另一优选例中，所述粘酸盐晶型D具有基本如图18所示的X-射线粉末衍射特征峰。
在另一优选例中，所述粘酸盐晶型D的差示扫描量热分析(DSC)图在加热至140.9±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述粘酸盐晶型D的差示扫描量热分析(DSC)图基本如图19所示。
在另一优选例中，所述粘酸盐晶型D的热重分析(TGA)图在加热至100℃时具有约1.45±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述粘酸盐晶型D的热重分析(TGA)图基本如图19所示。
代表性地，本发明提供一种式Ic-1化合物的富马酸盐晶型E，所述富马酸盐晶型E在1个或多个选自下组的2θ值处具有特征峰：4.7±0.2°、9.41±0.2°、13.34±0.2°、14.17±0.2°、15.63±0.2°、17.62,18.95±0.2°、22.75±0.2°、23.76±0.2°、25.66±0.2°、26.93±0.2°、28.69±0.2°、31.34±0.2°。
富马酸的结构式如下
Figure PCTCN2021076337-appb-000042
在另一优选例中，所述富马酸盐晶型E具有基本如图21所示的X-射线粉末衍射特征峰。
在另一优选例中，所述富马酸盐晶型E的差示扫描量热分析(DSC)图在加热至76.5±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述富马酸盐晶型E的差示扫描量热分析(DSC)图基本如图22所示。
在另一优选例中，所述富马酸盐晶型E的热重分析(TGA)图在加热至80℃时具有约2.17±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述富马酸盐晶型E的热重分析(TGA)图基本如图22所示。
代表性地，本发明提供一种式Ic-1化合物的D-葡萄糖醛酸盐晶型F，所述D-葡萄糖醛酸盐晶型F在1个或多个选自下组的2θ值处具有特征峰：4.77±0.2°、8.34±0.2°、10.87±0.2°、16.13±0.2°、17.54±0.2°、19.53±0.2°、20.06±0.2°、21.25±0.2°、23.42±0.2°、25.93±0.2°。
D-葡萄糖醛酸的结构式如下
Figure PCTCN2021076337-appb-000043
在另一优选例中，所述D-葡萄糖醛酸盐晶型F具有基本如图24所示的X-射线粉末衍射特征峰。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的差示扫描量热分析(DSC)图在加热至119.1±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的差示扫描量热分析(DSC)图基本如图25所示。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的热重分析(TGA)图在加热至100℃时具有约2.71±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失。
在另一优选例中，所述D-葡萄糖醛酸盐晶型F的热重分析(TGA)图基本如图25所示。
瞬时受体电位通道蛋白(TRP)
瞬时受体电位通道蛋白是一类存在于细胞膜上的重要阳离子通道构成的蛋白超家族。瞬时受体电位通道蛋白包括多个亚族，如TRPA、TRPC、TRPM、TRPV、TRPML和TRPP亚族。
研究发现，TRPA1通道蛋白与疼痛、癫痫、炎症、呼吸障碍、瘙痒、尿路障碍、炎症性肠病等疾病相关，TRPA1是治疗疼痛、癫痫、炎症、呼吸障碍、瘙痒、尿路障碍、炎症性肠病等疾病的靶标。
在本发明的一个优选例中，所述的瞬时受体电位通道蛋白(TRP)为TRPA1。
在另一优选例中，与瞬时受体电位通道蛋白(TRP)相关的疾病包括(但不限于)：疼痛、癫痫、炎症、呼吸障碍、瘙痒、尿路障碍、炎症性肠病，或其组合。
代表性地，与瞬时受体电位通道蛋白(TRP)相关的疾病为疼痛。
在另一优选例中，所述的疼痛为急性疼痛或慢性疼痛。
在另一优选例中，所述的疼痛包括(但不限于)：急性炎性疼痛、慢性炎性疼痛、内脏痛、神经源性疼痛、肌纤维痛、头痛、神经痛、癌症引起疼痛，或其组合。
在另一优选例中，所述的疼痛为炎症疼痛。
在另一优选例中，所述的炎性疼痛急性炎性疼痛或慢性炎性疼痛。
在另一优选例中，所述的头痛为偏头痛或肌紧张性疼痛。
在另一优选例中，所述的神经痛为三叉神经痛、糖尿病性疼痛或带状孢疹后神经痛。
在另一优选例中，所述的疼痛包括(但不限于)：急性疼痛、肌纤维痛、内脏疼痛、炎症疼痛、神经痛，或其组合。
在另一优选例中，所述的疼痛为肌纤维痛。
用途
本发明还提供了一种抑制瞬时受体电位通道蛋白(TPR)的方法，以及治疗与瞬时受体电位通道蛋白相关的疾病的方法。
本发明的上述式I化合物，或其药学上可接受的盐，以及式I化合物的盐酸盐晶型A、马来酸盐晶型B、草酸盐晶型C、粘酸盐晶型D、富马酸盐晶型E和D-葡萄糖醛酸盐晶型F可用于抑制瞬时受体电位通道蛋白，进而预防或治疗与瞬时受体电位通道蛋白相关的疾病。
在本发明中，与瞬时受体电位通道蛋白相关的疾病的例子包括(但并不限于)：疼痛、癫痫、炎症、呼吸障碍、瘙痒、尿路障碍、炎症性肠病。代表性的，所述的疼痛包括(但不限于)：急性炎性疼痛、慢性炎性疼痛、内脏痛、神经源性疼痛、肌纤维痛、头痛(例如偏头痛、肌紧张性疼痛等)、神经痛(例如三叉神经痛、糖尿病性疼痛、带状孢疹后神经痛等)、或癌症引起疼痛。
在一个优选实施例中，本发明提供了一种体外非治疗性的抑制瞬时受体电位通道蛋白活性的方法，包括例如在体外培养体系中，将瞬时受体电位通道蛋白或表达所述蛋白的细胞与本发明所述的式I化合物，或其药学上可接受的盐，或式I化合物的盐酸盐晶型A、马来酸盐晶型B、草酸盐晶型C、粘酸盐晶型D、富马酸盐晶型E或D-葡萄糖醛酸盐晶型F进行接触，从而抑制瞬时受体电位通道蛋白的活性。
本发明还提供了一种抑制瞬时受体电位通道蛋白的方法，该方法可以是治疗性的或非治疗性的。通常，该方法包括步骤：给需要的对象施用本发明所述的式I化合物，或其药学上可接受的盐、或式I化合物的盐酸盐晶型A、马来酸盐晶型B、草酸盐晶型C、粘酸盐晶型D、富马酸盐晶型E或D-葡萄糖醛酸盐晶型F。
优选地，所述对象包括人和非人哺乳动物(啮齿动物、兔、猴、家畜、狗、猫等)。
组合物和施用方法
本发明提供了一种用于抑制瞬时受体电位通道蛋白活性的组合物。
本发明所述的组合物包括(但并不限于)：药物组合物、食品组合物、膳食补充剂、饮料组合物等。
代表性地，所述的组合物为药物组合物，所述的药物组合物包括如本发明所述的式I化合物，或其药学上可接受的盐；和药学上可接受的载体。
代表性地，典型地，所述的组合物为药物组合物，所述的药物组合物包括如本发明所述的式I化合物式I化合物的盐酸盐晶型A、马来酸盐晶型B、草酸盐晶型C、粘酸盐晶型D、富马酸盐晶型E或D-葡萄糖醛酸盐晶型F；和药学上可接受的载体。
在本发明中，药物组合物的剂型包括(但不限于)口服制剂、注射剂、外用制剂。
代表性的包括(但不限于)：片剂、注射剂、输液剂、膏剂、凝胶剂、溶液剂、微 球、膜剂。
术语“药学上可接受的载体”指的是：一种或多种相容性固体、半固体、液体或凝胶填料，它们适合于人体或动物使用，而且必须有足够的纯度和足够低的毒性。“相容性”是指药物组合物中的各组分和药物的活性成分以及它们之间相互掺和，而不明显降低药效。
应理解，在本发明中，所述的载体没有特别的限制，可选用本领域常用材料，或用常规方法制得，或从市场购买得到。药学可接受的载体部分例子有纤维素及其衍生物(如甲基纤维素、乙基纤维素、羟丙甲基纤维素、羧甲基纤维素钠等)、明胶、滑石粉、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油、等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂(如吐温)、润湿剂(如十二烷基硫酸钠)、缓冲剂、螯合剂、增稠剂、pH调节剂、透皮促进剂、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、抑菌剂、无热原水等。
代表性的，液体剂型除了活性药物成分外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例如，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂等
药物制剂应与给药方式相匹配。本发明药剂还可与其他协同治疗剂一起使用(包括之前、之中或之后使用)。使用药物组合物或制剂时，是将安全有效量的药物施用于所需对象(如人或非人哺乳动物)，所述安全有效量通常至少约10微克/千克体重，而且在大多数情况下不超过约8毫克/千克体重，较佳地该剂量是约10微克/千克体重-约1毫克/千克体重。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。
本发明的主要优点包括：
(1)本发明提供了一类结构新颖且具有优异TRP通道抑制活性的式I化合物。本发明化合物具有优异的镇痛等体内药效、毒性更小、活性更高、安全窗口大、成药性好且具有优异的药代动力学性质。
(2)本发明还提供一种式I化合物和式I-a化合物的制备方法，所述的方法简单、易操作、收率和纯度高，且适合工业化生产。
(3)本发明还提供一种式Ic-1化合物的盐晶型，如式Ic-1化合物的盐酸盐晶型A、马来酸盐晶型B、草酸盐晶型C、粘酸盐晶型D、富马酸盐晶型E和D-葡萄糖醛酸盐晶型F，所述的式Ic-1化合物的盐晶型为固体形式，与游离的式Ic-1化合物油状物相比，固体形式的Ic-1化合物的盐晶型便于储存、运输和成药性强，且溶解度高，稳定性强(尤其是具有优异的热稳定性和高湿度稳定性)。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。
实施例1
式Ic-1化合物的制备
(1)(S)-3-(苯并呋喃-7-基氧基)-N,N-二甲基-3-(噻吩-2-基)丙-1-胺(Ic-8)的制备
Figure PCTCN2021076337-appb-000044
将1kg(7.35mol)7-氟苯并呋喃(式i化合物)室温下溶于8L二甲基亚砜溶液中，搅拌条件下加入2.04kg(11.02mol)的(S)-3-(二甲基氨基)-1-(噻吩-2-基)丙-1-醇(式ii化合物)和0.18kg(1.10mol)的碘化钾，反应瓶中通入氮气保护，冰水浴降温至10～20℃，往体系中加入1.47kg(36.13mol)氢氧化钠，加料完毕后将反应体系加热至50～60℃，在该温度下反应22小时，HPLC监测反应完毕后，用冰水浴降温至25～30℃，往体系中缓慢加入5L的水，控制体系温度低于45℃，加入5L乙酸乙酯萃取，水层再用2L乙酸乙酯洗涤，合并有机层，有机层用氯化钠饱和水溶液洗涤。将有机层中加入5L的水，搅拌条件下缓慢加入0.66kg的草酸，继续搅拌40分钟，静置分层10分钟，将有机层再加入1.70kg的10％的草酸水溶液，搅拌10分钟，分层，合并水相，将水相中加入5L乙酸乙酯，搅拌条件下往体系中加入饱和碳酸氢钠水溶液，调节pH至8，搅拌20分钟。过滤，滤饼用乙酸乙酯洗涤，滤液分层，收集有机相，水相用乙酸乙酯洗涤，合并有机相，无水硫酸钠干燥，浓缩至得1.12kg棕色油状目标产物式Ic-8化合物，收率50.6％，纯度95.6％。
1H NMR(400MHz,CDCl 3)δ7.62(d,J＝2.0Hz,1H),7.23(d,J＝5.0Hz,1H),7.17(d,J＝7.8Hz,1H),7.03(t,J＝7.9Hz,2H),6.96–6.87(m,1H),6.82(d,J＝7.9Hz,1H),6.74(d,J＝2.1Hz,1H),5.88–5.79(m,1H),2.60(t,J＝6.9Hz,2H),2.47(dt,J＝21.7,7.4Hz,1H),2.32(s,6H),2.22(dt,J＝20.5,6.8Hz,1H).MS(ESI,m/z):301.88(M+H) +.
(2)(S)-苯基(3-(苯并呋喃-7-氧基)-3-(噻吩-2-基)丙基)(甲基)氨基甲酸酯(I-a-1-1)的制备
Figure PCTCN2021076337-appb-000045
将1.12kg(3.65mol)的(S)-3-(苯并呋喃-7-基氧基)-N,N-二甲基-3-(噻吩-2-基)丙-1-胺(Ic-8)溶于5.5L的甲苯溶液中，搅拌条件下往体系中加入0.71kg(5.48mol)的N,N-二异丙基乙胺，通入氮气，用冰水浴降温将反应体系降至20～30℃，往体系中缓慢滴加0.86kg(5.48mol)氯甲酸苯酯，控制温度≤35℃，加完料将体系加热至40～45℃反应3小时。HPLC监测反应完毕，冰水浴降温至25～30℃。向反应体系中缓慢加入4.4L水，4.41L乙酸乙酯萃取，收集有机相，有机相用饱和氯化钠水溶液洗涤，浓缩，得到1.89kg黄色油状目标产物式iii化合物，收率127.1％，纯度80.5％。
1H NMR(500MHz,DMSO)δ7.91(t,J＝24.3Hz,1H),7.45(d,J＝5.0Hz,1H),7.39–7.26(m,2H),7.24–7.13(m,3H),7.06(dd,J＝23.8,15.4Hz,2H),6.98–6.85(m, 4H),5.93(d,J＝40.9Hz,1H),3.63(dd,J＝31.9,6.1Hz,1.5H),3.44(t,J＝23.1Hz,0.5H),2.96(t,J＝35.7Hz,3H),2.48–2.38(m,1H),2.26(dd,J＝60.2,26.5Hz,1H) ..MS(ESI,m/z):407.90(M+H) +。
(3)(S)-3-(苯并呋喃-7-基氧基)-N-甲基-3-(噻吩-2-基)丙-1-胺的制备(Ic-1)
Figure PCTCN2021076337-appb-000046
将1.89kg(4.63mol)(S)-苯基(3-(苯并呋喃-7-氧基)-3-(噻吩-2-基)丙基)(甲基)氨基甲酸酯(iii)加入7.54L的二甲基亚砜溶液中，搅拌条件下往体系中加入5.76kg质量分数为19.1％氢氧化钠水溶液，体系升温至60～65℃反应7小时。HPLC监测反应完毕，反应体系用冰水浴降温至15～20℃。向体系中缓慢加入水和乙酸乙酯萃取，有机相用饱和氯化钠水溶液洗涤后，往有机相中加入0.415kg(4.63mol)草酸，体系搅拌40分钟后过滤，滤饼用乙酸乙酯洗涤两次。将滤饼加入水，搅拌5分钟后过滤，滤饼用水洗涤一次，重复2次后，将滤饼加入乙酸乙酯和水，搅拌条件下往体系中滴加20～30％的氢氧化钠水溶液，调节pH至10～11，搅拌10分钟。将反应体系过滤，滤饼用乙酸乙酯洗涤两次，分层，水相用乙酸乙酯洗涤一次，合并有机相，有机相用饱和氯化钠水溶液洗涤，无水硫酸钠干燥，浓缩得0.73kg油状物目标产物Ic-1化合物，收率为54.9％，纯度98.3％
1H NMR(400MHz,CDCl 3)δ7.63(d,J＝2.0Hz,1H),7.20(t,J＝6.6Hz,2H),7.08–6.99(m,2H),6.88(dd,J＝4.9,3.6Hz,1H),6.80(d,J＝7.9Hz,1H),6.75(d,J＝2.0Hz,1H),5.93(dd,J＝8.2,4.4Hz,1H),3.30(t,J＝7.0Hz,2H),2.82–2.69(m,4H),2.65–2.54(m,1H).MS(ESI,m/z):287.87(M+H) +.
实施例2
(S)-7-(3-氯-1-(噻吩-2-基)丙氧基)苯并呋喃(中间体II-1)
Figure PCTCN2021076337-appb-000047
将480毫克(R)-3-氯-1-(噻吩-2-基)丙-1-醇，364毫克7-羟基苯并呋喃和784毫克三苯基膦溶解于20毫升的无水四氢呋喃中，在冰浴条件下慢慢往体系中滴加589微升偶氮二甲酸二异丙酯，滴加完毕将体系转移至室温反应过夜。反应完毕后，体系直接旋干，残余物经柱层析分离纯化，得到标题化合物，600毫克的无色油状，收率75.4％。
1H NMR(500MHz,CDCl 3)δ7.61(t,J＝3.1Hz,1H),7.39(dd,J＝1.7,0.7Hz,1H),7.21(dt,J＝8.2,1.9Hz,1H),7.11–7.06(m,1H),6.88(d,J＝7.9Hz,1H),6.76(dd,J＝8.1,2.1Hz,1H),6.33(d,J＝3.2Hz,1H),6.30(dd,J＝3.3,1.8Hz,1H),5.70(dd,J＝8.4,5.1Hz,1H),3.89(ddd,J＝11.1,8.2,5.4Hz,1H),3.73–3.65(m,1H),2.80–2.70(m,1H),2.51–2.42(m,1H).MS(ESI,m/z):293(M+H) +.
实施例3
(S)-3-(苯并呋喃-7-基氧基)-N-甲基-3-(噻吩-2-基)丙-1-胺(化合物I C-1)
Figure PCTCN2021076337-appb-000048
将600毫克中间体II-1溶于饱和碘化钠的丙酮溶液中，回流过夜。反应结束后，旋干溶剂，向体系中加入水，用乙酸乙酯萃取三次，饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩，残余物溶解于20毫升的四氢呋喃溶液中，加入2毫升40％的甲胺水溶液，反应过夜。反应结束后，旋干溶剂，向体系中加入氢氧化钠水溶液，用乙酸乙酯萃取三次，饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩，残余物经柱层析分离(甲醇/二氯甲烷＝1:15)，得到标题化合物，200毫克无色油状物，收率34.0％。
1H NMR(400MHz,CDCl 3)δ7.63(d,J＝2.0Hz,1H),7.20(t,J＝6.6Hz,2H),7.08–6.99(m,2H),6.88(dd,J＝4.9,3.6Hz,1H),6.80(d,J＝7.9Hz,1H),6.75(d,J＝2.0Hz,1H),5.93(dd,J＝8.2,4.4Hz,1H),3.30(t,J＝7.0Hz,2H),2.82–2.69(m,4H),2.65–2.54(m,1H).MS(ESI,m/z):287.87(M+H) +.
实施例4
(S)-3-(苯并呋喃-7-基氧基)-N-甲基-3-(噻吩-3-基)丙-1-胺(化合物I C-2)
Figure PCTCN2021076337-appb-000049
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-(噻吩-3-基)丙-1-醇外，其余所需原料、试剂及制备方法同实施例2-3，得到200毫克黄色油状物标题化合物，收率33.9％。 1H NMR(500MHz,CDCl 3)δ7.63(d,J＝2.1Hz,1H),7.27(dd,J＝5.0,3.0Hz,1H),7.24(d,J＝2.1Hz,1H),7.18–7.11(m,2H),7.01(t,J＝7.9Hz,1H),6.75(d,J＝2.1Hz,1H),6.72(d,J＝7.8Hz,1H),5.64(dd,J＝8.0,5.0Hz,1H),2.94–2.81(m,2H),2.48(s,3H),2.46–2.32(m,1H),2.24–2.13(m,1H).MS(ESI,m/z):287.76(M+H) +.
实施例5
(S)-3-(苯并呋喃-7-基氧基)-N-甲基-3-(呋喃-3-基)丙-1-胺(化合物I C-3)
Figure PCTCN2021076337-appb-000050
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-(呋喃-3-基)丙-1-醇外，其余所需原料、试剂及制备方法同实施例2-3，得到100毫克无色油状物标题化合物，收率9.9％。 1H NMR(500MHz,CDCl 3)δ7.62(d,J＝2.1Hz,1H),7.40(s,1H),7.35(t,J＝1.7Hz,1H),7.17(dd,J＝7.8,0.8Hz,1H),7.05(t,J＝7.9Hz,1H),6.80(d,J＝7.7Hz,1H),6.75(d,J＝2.1Hz,1H),6.46(d,J＝1.1Hz,1H),5.56(dd,J＝7.7,5.4Hz,1H),2.90–2.78(m,2H),2.46(s,3H),2.35(td,J＝13.9,7.4Hz,1H),2.13(dtd,J＝12.4,7.0,5.5Hz,1H).MS(ESI,m/z):271.88(M+H) +.
实施例6
(S)-3-(苯并呋喃-7-基氧基)-N-甲基-3-(呋喃-2-基)丙-1-胺(化合物I C-4)
Figure PCTCN2021076337-appb-000051
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-(呋喃-2-基)丙-1-醇外，其余所需原料、试剂及制备方法同实施例2-3，得到30毫克无色油状物标题化合物，收率3.2％。 1H NMR(500MHz,CDCl 3)δ7.61(d,J＝2.1Hz,1H),7.36(d,J＝1.1Hz,1H),7.20(d,J＝7.7Hz,1H),7.06(dd,J＝10.5,5.3Hz,1H),6.82(d,J＝7.7Hz,1H),6.74(dd,J＝7.8,2.1Hz,1H),6.31(d,J＝3.2Hz,1H),6.27(dd,J＝3.2,1.8Hz,1H),5.60(dd,J＝7.9,5.3Hz,1H),3.13–2.99(m,2H),2.63–2.56(m,4H),2.44(ddd,J＝14.1,12.4,7.0Hz,1H).MS(ESI,m/z):271.88(M+H) +.
实施例7
(S)-3-(2,3-二氢-1H-茚-4-基)氧基)-N-甲基-3-(2-噻吩基)丙-1-胺(化合物I C-10)
Figure PCTCN2021076337-appb-000052
除了将7-羟基苯并呋喃换成4-茚醇外，其余所需原料、试剂及制备方法同实施例2-3，得到16毫克棕色油状标题化合物，收率是9.9％。
1H NMR(400MHz,CDCl 3)δ7.21(dd,J＝5.0,1.2Hz,1H),7.03–6.96(m,2H),6.92(dd,J＝5.0,3.5Hz,1H),6.81(d,J＝7.3Hz,1H),6.65(d,J＝8.0Hz,1H),5.57(dd,J＝7.8,5.0Hz,1H),2.87(dt,J＝19.9,7.3Hz,6H),2.47(s,3H),2.41–2.30(m,1H),2.25–2.14(m,1H),2.06(p,J＝7.0Hz,2H).MS(ESI,m/z):287.87(M+H) +.
实施例8
(S)-2-(3-(苯并呋喃-7-基氧基)-3-(噻吩-2-基)丙基)异吲哚啉-1,3-二酮(中间体III-1)
Figure PCTCN2021076337-appb-000053
将160毫克中间体II-1，303毫克的邻苯二甲酰亚胺钾盐和25毫克的碘化钠溶解于5毫升的N,N-二甲基甲酰胺中，氮气保护条件下90℃反应过夜。反应结束后，向体系中加入水，用乙酸乙酯萃取三次，水洗，饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩，残余物经柱层析分离(乙酸乙酯/石油醚＝1:5)，得到标题化合物，140毫克黄色的固体，收率63.5％。
1H NMR(500MHz,CDCl 3)δ7.81–7.78(m,2H),7.68(dd,J＝5.5,3.0Hz,2H),7.45(d,J＝2.1Hz,1H),7.18(dd,J＝5.0,1.1Hz,1H),7.14(dd,J＝7.8,0.8Hz,1H),7.06(d,J＝3.0Hz,1H),7.04–6.97(m,1H),6.86(dt,J＝10.3,5.2Hz,1H),6.78(d,J＝7.4Hz,1H),6.68(d,J＝2.1Hz,1H),5.82(dd,J＝7.8,5.2Hz,1H),4.07–3.89(m,2H),2.66(td,J＝14.4,7.3Hz,1H),2.47–2.36(m,1H).MS(ESI,m/z):404(M+H) +.
实施例9
(S)-3-(苯并呋喃-7-基氧基)-3-(噻吩-2-基)丙-1-胺(化合物I C-23)
Figure PCTCN2021076337-appb-000054
将140毫克中间体III-1和87毫克的水合肼溶解于5毫升的甲醇溶液中，室温反应过夜。反应结束后，旋干溶剂，残余物经柱层析分离(甲醇/二氯甲烷＝1:15)，得到标题化合物，30毫克无色油状物，收率31.6％。
1H NMR(500MHz,DMSO)δ7.97(d,J＝2.1Hz,1H),7.49(dd,J＝5.0,1.1Hz,1H),7.24–7.17(m,2H),7.11–7.03(m,1H),6.99(dd,J＝5.0,3.5Hz,1H),6.95(d,J＝7.5Hz,1H),6.93(d,J＝2.1Hz,1H),6.04(dd,J＝7.8,5.5Hz,1H),2.99–2.86(m,2H),2.44–2.35(m,1H),2.21(ddt,J＝11.5,9.3,5.8Hz,1H).MS(ESI,m/z):273.77(M+H) +.
实施例10
(S)-3-(苯并呋喃-7-基氧基)-3-(呋喃-3-基)丙-1-胺(化合物I C-24)
Figure PCTCN2021076337-appb-000055
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-(呋喃-3-基)丙-1-醇外，其余所需原料、试剂及制备方法同实施例2、8-9，得到90毫克无色油状物标题化合物，收率是12.7％。
1H NMR(500MHz,CDCl 3)δ7.62(d,J＝2.1Hz,1H),7.42–7.37(m,1H),7.35(t,J＝1.7Hz,1H),7.18(dd,J＝7.8,0.9Hz,1H),7.08–7.01(m,1H),6.79(dd,J＝7.9,0.7Hz,1H),6.75(d,J＝2.2Hz,1H),6.45(dd,J＝1.7,0.7Hz,1H),5.57(dd,J＝8.1,5.0Hz,1H),3.03–2.91(m,2H),2.31(ddt,J＝14.1,8.1,6.4Hz,1H),2.06(dtd,J＝9.4,7.1,5.1Hz,1H).MS(ESI,m/z):257.77(M+H) +.
实施例11
(S)-3-(苯并呋喃-7-基氧基)-3-(呋喃-2-基)丙-1-胺(化合物I C-25)
Figure PCTCN2021076337-appb-000056
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-(呋喃-2-基)丙-1-醇外，其余所需原料、试剂及制备方法同实施例2、8-9，得到85毫克无色油状物标题化合物，收率是16.5％。
1H NMR(500MHz,CDCl3)δ7.62(d,J＝2.1Hz,1H),7.37(dd,J＝1.6,1.0Hz,1H),7.20(dd,J＝7.8,0.9Hz,1H),7.06(t,J＝7.9Hz,1H),6.87–6.80(m,1H),6.75(d,J＝2.1Hz,1H),6.33–6.24(m,2H),5.59(dd,J＝8.0,5.5Hz,1H),3.09–2.93(m,2H),2.43(dq,J＝7.9,6.4Hz,1H),2.22(qd,J＝12.5,6.9Hz,1H).MS(ESI,m/z):257.64(M+H) +.
实施例12
(S)-3(苯并呋喃-7-基氧基)-3-(噻吩-3-基)丙-1-胺(化合物I C-26)
Figure PCTCN2021076337-appb-000057
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-(噻吩-3-基)丙-1-醇外，其余所需原料、试剂及制备方法同实施例2、8-9，得到60毫克黄色油状物标题化合物，收率是14.6％。
1H NMR(500MHz,CDCl3)δ7.62(d,J＝1.9Hz,1H),7.25–7.22(m,2H),7.13(t,J＝7.4Hz,1H),7.09(d,J＝4.6Hz,1H),6.97(t,J＝7.9Hz,1H),6.71(d,J＝2.0Hz,1H),6.67(d,J＝8.0Hz,1H),5.65(dd,J＝7.9,4.2Hz,1H),3.30–3.11(m,2H),2.49(dd,J＝14.1,7.4Hz,1H),2.34(dd,J＝12.9,5.8Hz,1H).MS(ESI,m/z):273.77(M+H) +.
实施例13
(S)-3-(苯并呋喃-7-基氧基)-N-甲基-3-(噁唑-5-基)丙-1-胺(化合物I C-29)
Figure PCTCN2021076337-appb-000058
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-(噁唑-5-基)丙-1-醇外，其余所需原料、试剂及制备方法同实施例2-3，得到63毫克黄色油状标题化合物，收率是21.0％。1H NMR(500MHz,CDCl3)δ7.83(s,1H),7.62(d,J＝2.1Hz,1H),7.23(dd,J＝7.8,0.9Hz,1H),7.08(dd,J＝10.4,5.3Hz,1H),7.03(s,1H),6.87–6.81(m,1H),6.76(d,J＝2.2Hz,1H),5.74(dd,J＝8.0,5.4Hz,1H),3.00–2.87(m,2H),2.53–2.49(m,4H),2.36–2.26(m,1H).MS(ESI,m/z):272.76(M+H) +.
对比例1
提供如下式C1化合物的简要制备
Figure PCTCN2021076337-appb-000059
除了将(R)-3-氯-1-(噻吩-2-基)丙-1-醇换成(R)-3-氯-1-苯基丙烷-1-醇外，其余所需原料、试剂及制备方法同实施例7，得到14毫克化合物C1，收率是18.1％。
1H NMR(400MHz,CDCl 3)δ7.46–7.14(m,5H),6.90(t,J＝7.7Hz,1H),6.77(d,J＝7.4Hz,1H),6.41(d,J＝8.1Hz,1H),5.33(dd,J＝8.0,4.4Hz,1H),3.13(dd,J＝13.5,6.0Hz,2H),2.91(dt,J＝12.3,7.5Hz,4H),2.62(s,3H),2.52–2.36(m,2H),2.13–2.00(m,2H).MS(ESI,m/z):282.20(M+H) +.
实施例14
TRPA1的抑制活性
在本实施例中，测试本发明部分实施例化合物对瞬时受体电位通道蛋白TRPA1的抑制活性。其中，阳性对照化合物采用式A化合物(WO2010075353)：
Figure PCTCN2021076337-appb-000060
式A化合物
方法如下：
通过IonWorks Barracuda(IWB)自动化膜片钳检测的测试方法：稳定表达TRPA1的HEK293细胞，用含有15μg/mL Blasticidin S HCl、200μg/mL Hygromycin B和10％FBS血清的DMEM培养基，置于T175的培养瓶中，放入37℃，5％CO 2的培养箱中培养，待细胞密度生长到～80％时，移走培养液，用无钙镁的磷酸缓冲液(PBS)冲洗一遍，加入3mL的Trypsin消化2分钟，加入7mL培养液终止消化。将细胞收集到15mL的离心管中以800转/每分钟离心3分钟，去除上清液后将细胞加入适当体积的细胞外液重悬，使细胞密度控制在2-3×10 6/mL并用于IWB实验。细胞外液配方(in mM)：140NaCl,5KCl,1MgCl 2,10HEPES,0.5EGTA,10Glucose(pH 7.4)；细胞内液配方(in mM)：140CsCl,10HEPES,5EGTA,0.1CaCl 2,1MgCl 2(pH 7.2)。两性霉素B与实验当天用DMSO新鲜配制成28mg/mL，再用细胞内液配制成0.1mg/mL的终浓度。
IWB实验使用population patch clamp(PPC)板，全部检测过程由仪器自动完成，即在PPC板的384孔中加入细胞外液，并在PPC板下即plenum内加入细胞内液后，加入6L的细胞液进行封接测试，最后将plenum中的细胞内液换成含两性霉素B的细胞内液，使封接的细胞穿孔后形成全细胞记录模式。记录TPRA1电流的采样频率为10kHz，细胞钳制在0mV，电压刺激命令(channel protocol)为一个300ms从-100mV到+100mV的斜坡(ramp)电压，每10s给予此电压刺激，mTRPA1电流由300M AITC诱发。
数据记录和电流幅度测量导出由IWB软件完成(version 2.5.3，Molecular Devices Corporation，Union City，CA)。封接阻抗低于20MΩ的孔将不记录数据统计。原始电流数据由软件进行漏减矫正，TRPA1电流幅度在+100mV时测得。实验的每块PPC板都将有一个HC030031的剂量效应数据作为阳性对照，如HC030031的IC 50值超过以往每块板上得到的IC 50平均值的3倍时，将进行复测。化合物剂量效应曲线和IC 50由GraphPad Prism 5.02(GraphPad Software,San Diego,CA)进行拟合计算。
实验结果
对本发明的部分化合物通过IonWorks Barracuda(IWB)自动化膜片钳检测的测试方法，进行IC 50抑制活性测试，活性数据如表2所示，部分代表性化合物的抑制TRPA1活性的量效关系如图1A-1E所示。
表2.本发明的部分化合物在自动化膜片钳检测测试中对TRPA1的抑制活性数据(IC 50,μM)
Figure PCTCN2021076337-appb-000061
Figure PCTCN2021076337-appb-000062
其中活性：IC 50(μM)：
50-100：+
20-50：++
10-20：+++
5-10：++++
1-5：+++++
结果表明，本发明的化合物对TRPA1表现出强效抑制活性，其中有11个化合物对TRPA1的半数有效抑制浓度IC 50在1-10μM之间，从图1A-1E中可以看出，Ic-3、Ic-4、Ic-8、Ic-23、Ic-24化合物对TRPA1的半数有效抑制浓度IC 50<5μM，因此，可以得出本发明式I化合物对TRPA1具有很强的抑制活性。
此外，化合物I C-10(含有杂芳基
Figure PCTCN2021076337-appb-000063
)和化合物C1(含有苯基)的活性之比(化合物C1的IC 50/化合物I C-10的IC 50)为约2.5倍，这表明，含有杂芳基的本发明化合物(如I C-10)对TRPA1的抑制活性更高。
此外，与A基团为萘环的化合物(如度洛西汀)相比，化合物I C-10(A基团为苯并脂肪环)以及化合物I C-3、化合物I C-23和化合物I C-1(其中A基团均为苯并杂芳基)的IC 50值显著降低。具体地，S型度洛西汀的IC 50与化合物I C-10、化合物I C-3、化合物I C-23或化合物I C-1中任一化合物的IC 50的比值为约2.8～6.8。这表明，A基团为苯并脂肪环或杂芳基的本发明化合物，对TRPA1的抑制活性更高(提高了约2.8-6.8倍)。
类似地，本发明人还对S型度洛西汀和化合物I C-10，通过手动膜片钳测试法测定了TRPA1抑制活性。与自动膜片钳测试法的测试结果类似，在手动膜片钳测试法中，S型度洛西汀的IC 50与化合物I C-10的IC 50的比值为4.36/1.12＝3.9。
类似地，本发明人还对化合物I C-1，通过手动膜片钳测试法测定了TRPA1抑制活性，测定方法如下：
稳定表达人TRPA1通道的HEK293稳转细胞株，用含有15μg/mL Blasticidin S HCl、200μg/mL Hygromycin B和10％FBS血清的DMEM培养基，置于T75的培养瓶中，放入37℃，5％CO2的培养箱中培养，待细胞密度生长到～80％时，移走培养液，用无钙镁的磷酸缓冲液(PBS)冲洗一遍，加入2mL的Trypsin消化2分钟，加入8mL培养液终止消化。将细胞收集到15mL的离心管中以800转/每分钟离心3分钟，去除上清液后将细胞加入适当体积的细胞外液重悬。
手动膜片钳检测是在室温条件下，采用HEKA系统(Patch Master软件)结合EPC-10放大器记录TRPA1稳转细胞株的全细胞电流。全细胞记录用内液配方(mM)：140CsCl,10HEPES,5EGTA,0.1CaCl2,1MgCl2(pH7.2，渗透压295–300mOsm)；记录用外液采用无Ca2+设置(mM)：140NaCl,5KCl,0.5EGTA,1MgCl2,10Glucose,10HEPES(pH 7.4，渗透压300–310mOsm)。膜片钳记录使用的玻璃微电极电阻2-4MΩ，采样频率10kHz，滤波频率2.9kHz，细胞钳制在0mV，电压刺激命令(channel protocol)为一个300ms从-100mV到+100mV的线性电压，之后恢复至0mV的钳制电位，此记录每2s进行一次，hTRPA1电流由100μM AITC诱发，为确保电流记录的准确，记录时串联 电阻进行60％的补偿。
数据记录和电流幅度测量导出由Patch Master软件完成。封接阻抗低于500MΩ的细胞将不纳入数据统计。原始电流数据由软件进行漏减矫正，hTRPA1电流幅度在+100mV时测得。化合物剂量效应曲线和IC50由GraphPad Prism 5.02(GraphPad Software,San Diego,CA)进行拟合计算。
手动膜片钳测试结果
与自动膜片钳测试法的测试结果类似，在手动膜片钳测试法中，S型度洛西汀的IC 50与化合物I C-1的IC 50的比值为4.36μM/0.43μM＝10.14。
实施例15
细胞毒性试验
在本实施例中，测定实施例制备的I C-10和I C-1化合物的肝细胞毒性和神经细胞毒性实验，方法如下：
准备HepG-2和SH-SY5Y细胞，10cmdish中于37℃，5％CO 2细胞培养箱内培养；胰酶消化重悬细胞并计数，按100μl/孔的体系，8000cells的量将细胞转接至96孔板中。37℃，5％CO 2细胞培养箱内培养24小时；准备化合物梯度浓度体系，2倍稀释，体系为100μl/孔。去掉第一天中96孔板细胞培养体系中的上清，并将新配置好的药物浓度体系对应加入到培养细胞的培养板孔内(设置双复孔)。37℃，5％CO 2细胞培养箱内培养72h。细胞培养结束之后，去掉96孔板细胞培养体系中的上清，每孔加入100μl检测溶液(含10％CCK-8的培养基)，37℃，5％CO 2细胞培养箱内孵育1h，到时取出用酶标仪测定在450nm处的吸光度。进行数据处理，计算细胞毒性，并用GraphPad Prism计算IC50，细胞毒性计算公式如下：细胞毒性(％)＝[A(0加药)-A(加药)]/[A(0加药)-A(空白)]×100
A(加药)：具有细胞、CCK-8溶液和药物溶液的孔的吸光度
A(空白)：具有培养基和CCK-8溶液而没有细胞的孔的吸光度
A(0加药)：具有细胞、CCK-8溶液而没有药物溶液的孔的吸光度
实验结果
I C-10和I C-1化合物的肝细胞毒性(HepG2细胞)和神经细胞(SH-SY5Y)毒性的结果表明：
度洛西汀对肝细胞毒性和神经细胞毒性(IC 50,μM)分别为33μM和28μM，而本发明的I C-1和I C-10化合物对肝细胞毒性和神经细胞毒性(IC 50,μM)为约为60-120μM，这表明，本发明化合物的毒副作用显著更低，化合物仅为度洛西汀毒副作用的约1/2或1/3)。这提示，本发明化合物具有优良的安全性。
实施例16
镇痛活性测试实验
在本实施例中，通过小鼠福尔马林疼痛模型测试本发明实施例制备的化合物I C-10、化合物的镇痛活性实验，方法如下：
取C57BL/6小鼠(雄性，9周)30只，小鼠随机分为3组：分别为溶剂对照组(vehicle，生理盐水)、度洛西汀组(Duloxetine，5-HT重摄取和NE重摄取抑制剂)和Ic-10组(本发明化合物Ic-10)。实验开始前，让小鼠适应实验环境72h，期间无需禁食禁水。采用腹腔注射方式给予受试药物，剂量为20mg/kg，然后将小鼠置于透明、通气的有机玻璃圆筒内1h后，于各组小鼠左后足底以微量进样器注射4％福尔马林溶液20μl，以微型摄像机实时记录小鼠的足部疼痛反应。以小鼠抬(1分/次)、抖(2分/次)和舔(3分/次)左足的次数以及舔左足的时间长度作为疼痛反应的指标，分别观察记录0-10min(I相， 急性疼痛期)和10-60min(II相，炎性疼痛期)两时段内的累积打分和舔足时间，并进行统计学分析。
实验结果
本发明的化合物I C-10在小鼠福尔马林疼痛模型中的镇痛活性结果如图2所示。从结果可知，在添足时间统计检测指标中,在20mg/kg给药剂量下，本发明的化合物I C-10在I相(0-10min)和II相(10-60min)均表现出明确并且强效的镇痛活性，与生理盐水组相比，几乎完全抑制了小鼠由于疼痛导致的添足行为，并且与临床用药度洛西汀镇痛活性相当。
实施例17
小鼠热板痛测试实验
在本实施例中，通过C57小鼠热板致痛模型测试本发明实施例制备的化合物I C-23、Ic-10、Ic-1等化合物镇痛活性实验，方法如下：
1动物筛选
取SPF级C57雄性小鼠，调节热板温度恒定于55±0.1℃，筛选10-30s内有舔足等疼痛反应的小鼠(对逃避和跳跃者弃之)。发现有疼痛反应立即取出，防止小鼠烫伤。
2动物分组
将筛选到的40只动物进行称重，并将动物随机平均分为4组，分别为生理盐水对照组(空白对照)、度洛西汀组(阳性对照组)、加巴喷丁组(阳性对照组)和Ic-23组(本发明化合物)。
3样品配置
受试化合物于给药当天新鲜配制。配置0.9％NaCl生理盐水溶液作为溶媒备用，将适量的受试化合物加入所需体积的生理盐水中，充分混悬，配制药物浓度为1mg/ml。小鼠的给药体积标准为10ml/kg(即0.1ml/10g)。
4动物给药
腹腔给药，给药前动物无需禁食禁水。给药体积为10ml/kg。度洛西汀和I C-23的给药剂量为10mg/kg,加巴喷丁的给药剂量为100mg/kg。
5热板实验观察
热板观察指标：小鼠在55±0.1℃的热板上的反应时间(Time latency)。给药前3h及给药后15min，30min，60min各测一次并记录。
6.数据统计和分析
使用最大可能镇痛效应％(maximum possible effect，MPE)来评判各化合物的镇痛效应，即MPE％＝[(Post drug latency-baseline latency)/(30-baseline latency)]×100。统计不同时间点的MPE％。MPE％的数值越大，说明化合物的镇痛药效越强。
实验结果
本发明的化合物I C-23在小鼠热板致痛模型中的镇痛活性结果如图3所示。从结果可知，与生理盐水对照组相比，本发明的化合物I C-23在10mg/kg的给药剂量下表现出非常强效的镇痛效果，具有显著性差异。与阳性对照组相比，在60分钟内本发明的化合物I C-23镇痛活性要明显优于100mg/kg的加巴喷丁，并且优于10mg/kg的度洛西汀镇痛效果。
此外，在相同剂量(10mg/kg)下，化合物Ic-10和Ic-1的镇痛活性均优于加巴喷丁和度洛西汀。
热板致痛模型是经典的评价药物急性痛药效的模型，因此本发明的化合物对急性疼痛具有优异的治疗效果。
实施例18
Ic-1药代动力学测试
在本实施例中，测试了度洛西汀和实施例1制备的Ic-1等化合物的大鼠药代动力学性质，方法如下：
测试方法：
称取一定量样品溶于去离子水中，配置浓度为1mg/mL的溶液。雄性SD大鼠作为测试动物。单次静脉(IV)注射剂量为2mg/kg，口服(PO)给药剂量为10mg/kg，每组设置三只大鼠。口服组给药前禁食10-14h。给药后4h后恢复给食。动物采血时间点为：静脉，给药前，给药后5，15，30min，1，2，4，6，8h和24h；口服，给药前，给药后15，30min，1，2，4，6，8和24h。每只动物每次经颈静脉采约0.25mL血液，肝素钠抗凝。血液样本采集后置于冰上，离心分离血浆(离心条件：8000转/分钟，6分钟，4℃)。收集的血浆分析前存放于-80℃。取50μL血浆样品至1.5mL离心管中，加入250μL内标溶液(空白组不加内标溶液，补加相同体积的甲醇)，涡旋混匀，14000转/分钟离心5min，取200μL上清液加入到96孔进样板中，LC-MS/MS进样分析。线性回归分析是以峰面积为y轴，以药物浓度为x轴。峰面积比和浓度之间的线性关系用由化合物的回归方程所得的相关系数(R)来表示。根据药物的血药浓度数据，使用药代动力学计算软件WinNonlin7.0非房室模型分别计算受试物的药代动力学参数。
内标工作液：吸取一定量的浓度为490,000ng/mL的甲苯磺丁脲内标储备液至一定体积的容量瓶中，用甲醇定容至刻度后混匀，制得浓度为200ng/mL的内标工作溶液。
实验结果
根据药物各组的平均血药浓度数据，使用药代动力学计算软件WinNonlin7.0非房室模型分别计算化合物各组的药代动力学参数，见表3。
表3 化合物I C-1和度洛西汀主要药代动力学参数
Figure PCTCN2021076337-appb-000064
从表3可以看出，I C-1以2mg/kg的给药剂量静脉注射后，取样第一个时间点0.083h即至峰浓度(C max,170ng/mL)，消除半衰期(T 1/2)为2.97h，AUC(0-∞)为449h*ng/mL；以5倍剂量(10mg/kg)灌胃口服给药后，3h达到峰浓度(C max，266ng/mL)，消除半衰期(T 1/2)为5.69h，AUC(0-∞)为4016h*ng/mL。以AUC(0-∞)计算，口服生物利用度为179％。
度洛西汀以2mg/kg的给药剂量静脉注射后，0.083h即至峰浓度(C max,177ng/mL)，消除半衰期(T 1/2)为1.77h，AUC(0-∞)为449h*ng/mL；以5倍剂量(10mg/kg)灌胃口服给药后，0.83h达到峰浓度(C max，76ng/mL)，消除半衰期(T 1/2)为1.81h，AUC(0-∞)为222h*ng/mL。以AUC(0-∞)计算，口服生物利用度为9.9％。
由以上结果可以看出，具有式I结构的本发明化合物(如化合物I C-1)相较于度洛西汀具有更优良的药代动力学性质，其半衰期更长，在血浆中暴露量更高，生物利用度更好，适合开发成口服给药，具有良好的成药前景。
实施例19
通过小鼠ICS模型考察实施例1制备的化合物Ic-1对肌纤维痛的治疗作用
实验方法
1、实验分组
实验分组为：溶剂对照组、10mg/kg度洛西汀组(阳性对照组)和10mg/kg化合物Ic-1组(实施例1制备化合物)
2、化合物配置
10mg/kg度洛西汀：称取17mg度洛西汀，使用生理盐水溶解并定容至8.5mL，充分溶解后口服灌胃给药，给药体积为5ml/kg。
10mg/kg Ic-1：称取17mg Ic-1，使用生理盐水溶解并定容至8.5mL，充分溶解后口服灌胃给药，给药体积为5ml/kg。
3、动物
取用雄性C57BL/6小鼠，实验开始时体重为18-22克，每笼分配4只小鼠，可自由采食和饮水。每个实验组12只小鼠，采用动物尾部标记法标记实验小鼠。
4、实验方法
4.1建立模型
第0天：下午4:30，将小鼠放入带有不锈钢网的有机玻璃盒里，然后有机玻璃盒放入冷库(温度4±2℃)中过夜。自由采食并用琼脂块代替水。
第1天：早上10:00，将小鼠转移到室温(温度24±2℃)环境下,放置30分钟，之后再转移到冷库中放置30分钟。重复上述步骤直到下午4:30，将小鼠放入冷库中过夜。
第2天：重复第1天的操作。
第3天：上午10:00将小鼠从冷库中转移出来。
4.2给药
化合物按照实验安排的时间口服给药，给药剂量为10mg/kg。
4.3机械痛觉超敏测试
造模后第四天，对动物进行给药前机械痛觉超敏测试，PWT值大于0.5g的动物将被剔除，不用于正式实验。造模后第五天，化合物给药后0.5小时、1小时、2小时，对动物进行机械痛觉超敏测试。
机械痛觉超敏测试方法如下：
将小鼠单独放置在有机玻璃盒中，盒子底部为网格以保证小鼠脚部可以测试。在测试前小鼠将适应15分钟。适应完成后，使用测试纤维在小鼠左后脚脚底中心部位测试。测试纤维包括8个测试强度：2.36(0.02g),2.44(0.04g),2.83(0.07g),3.22(0.16g),3.61(0.4g),3.84(0.6g),4.08(1g),4.17(1.4g)。测试时，将测试纤维垂直压向皮肤并施力使纤维弯曲6-8秒，每次测试间隔5秒。测试时，动物迅速缩脚被记为疼痛反应。测试纤维离开动物皮肤时动物缩脚也被记为疼痛反应。如果动物移动或走动，不记疼痛反应，应重复测试。测试时首先使用3.22(0.16g)，如果动物有疼痛反应，下一次测试使用力度小一级的测试纤维；如果动物没有疼痛反应，下一次测试使用力度大一级的测试纤维(Chaplan et al.1994)。测试纤维的最大力度为4.17(1.4g)。
测试结果记录在如下的表4中，有疼痛反应记录X，没有疼痛反应记录O。
表4
2.36      
2.44      
2.83      
3.22 O    
3.61 O O O
3.84 X X X
4.08      
4.17      
机械痛觉过敏使用如下公式计算：
50％反应阈值(g)＝(10 (Xf+k))/10,000
Xf＝测试中使用的最终测试纤维值
k＝表值(Chaplan et al.1994,page 62)
＝平均差
5、数据收集和分析
Figure PCTCN2021076337-appb-000065
化合物Ic-1在小鼠ICS模型中的镇痛活性结果如表5和图4所示。
表5 化合物Ic-1和度洛西汀在小鼠ICS模型中给药后不同时间机械痛觉超敏测试结果(PWT)统计数据
Figure PCTCN2021076337-appb-000066
从表5和图4中可以看出，本发明的化合物Ic-1在10mg/kg的给药剂量下表现出非常强效的镇痛效果，口服给药1小时和2小时都显示能抑制ICS模型引起的机械痛觉超敏。与阳性对照组相比，化合物Ic-1在0.5h、1h和2h三个测试时间下，镇痛效果全部优于度洛西汀。小鼠ICS模型是经典的评价药物治疗肌纤维痛的药效模型，因此本发明的化合物Ic-1对肌纤维疼痛具有优异的治疗效果。
实施例20
通过小鼠醋酸扭体痛模型考察实施例1制备的化合物I C-1对内脏疼痛、炎症疼痛的治疗作用
实验方法
取ICR小鼠，雄性，22-25g，给药前禁食2h，不禁水。对所有ICR小鼠进行称重，并随机分组，每组动物数>10只。阴性对照组为生理盐水组(Vehicle，空白对照)，阳性对照组设置为给药剂量10mg/kg吲哚美辛(一种非甾体类抗炎药)，给药剂量10mg/kg山莨菪碱(一种解痉药，临床上具有镇痛活性)，给药剂量10mg/kg和20mg/kg的度洛西汀。测试化合物为I C-1(实施例1制备的化合物)，给药剂量设置为5mg/kg和10mg/kg。根据小鼠体重通过灌胃的方式给药。给药后1h腹腔注射1.5％醋酸溶液(0.1ml/10g)，并 观察之后30min内各组小鼠出现内脏痛的次数，小鼠出现腹部内凹，躯干与后腿伸张，臀部高起记为一次，最终统计30min内出现上述现象的次数。给药后小鼠出现的内脏痛次数越少，说明化合物的镇痛药效越强。
实验结果
小鼠醋酸扭体痛模型测试如图5所示，从图5可以看出，本发明的化合物I C-1(5mg/kg和10mg/kg)单次灌胃给药可显著降低醋酸引起的小鼠扭体反应次数，与生理盐水组(Vehicle，空白对照)(49次)相比具有显著性差异。化合物I C-1在5mg/kg的给药剂量下，小鼠扭体反应次数为20次，低于生理盐对照组的49次的50％，提示化合物I C-1在该模型的半数有效剂量(ED 50)小于5mg/kg。10mg/kg剂量下的化合物I C-1镇痛效果(17次)要优于同剂量下的阳性药吲哚美辛(28次)、山莨菪碱(27次)和度洛西汀(27次)，并且5mg/kg剂量下化合物I C-1的镇痛效果(20次)与20mg/kg的度洛西汀的镇痛效果相当(21次)。该实验表明，在小鼠醋酸扭体痛模型中，本发明的化合物I C-1的镇痛活性明显优于阳性对照药物。小鼠醋酸扭体痛模型是经典的评价药物治疗内脏疼痛和炎症疼痛的药效模型，因此本发明的化合物I C-1对内脏疼痛和炎症疼痛具有优异的治疗效果。
实施例21
通过大鼠SNL模型考察实施例1制备的化合物I C-1对神经疼痛的治疗作用
实验方法
取SD大鼠进行手术，雄性，SPF级，质量150g-180g。手术过程执行无菌操作。使用戊巴比妥钠(50mg/kg,腹腔注射)麻醉动物。对动物腰部手术区域剃毛，使用碘伏和70％乙醇对皮肤消毒三遍。待皮肤干燥后开始手术。使用手术刀在动物腰部荐骨后部开一纵向切口，暴露左侧椎旁肌，使用撑开器分离肌肉组织以暴露脊椎骨。分离左侧脊神经L5和L6，使用6-0丝线结扎，缝合伤口。手术后将动物放置在电热毯上，皮下注射5mL生理盐水以防止脱水。等动物完全苏醒后(可自由活动)将动物放回笼中。
手术后，将动物放在实验环境中适应，15分钟/天，适应3天。给药前一天，对大鼠进行机械痛觉超敏基线测试，将没有表现出机械痛觉超敏的动物(缩脚阈值大于5g)的动物剔除后随机分成1组对照组和2组实验组。
称量动物体重，以给药剂量计，2组实验组分别通过灌胃给予100mg/kg加巴喷丁(加巴喷丁是目前临床上治疗神经痛的一线用药)和10mg/kg化合物I C-1(实施例1制备的化合物)，对照组为口服灌胃等体积的生理盐水。给药后1小时，进行机械痛觉超敏测试。将大鼠单独放置在有机玻璃盒中，盒子底部为网格以保证大鼠脚部可以测试。在测试前大鼠将适应15分钟。适应完成后，使用测试纤维在大鼠左后脚脚底中心部位测试。测试纤维包括8个测试强度：3.61(0.4g)，3.84(0.6g)，4.08(1g)，4.31(2g)，4.56(4g)，4.74(6g)，4.93(8g)，5.18(15g)。测试时，将测试纤维垂直压向皮肤并施力使纤维弯曲6-8秒，每次测试间隔5秒。测试时，动物迅速缩脚被记为疼痛反应。测试纤维离开动物皮肤时动物缩脚也被记为疼痛反应。如果动物移动或走动，不记疼痛反应，应重复测试。测试时首先使用4.31(2g)，如果动物有疼痛反应，下一次测试使用力度小一级的测试纤维；如果动物没有疼痛反应，下一次测试使用力度大一级的测试纤维)。测试纤维的最大力度为5.18(15g)。
机械痛觉超敏在大鼠行为学测试中表述为缩脚阈值(PWT)，按照如下公式计算：
50％反应阈值(g)＝(10 (Xf+k))/10,000
Xf＝测试中使用的最终测试纤维值
k＝表值
＝平均差
使用Excel软件收集数据，使用Prism 6.01(Graph pad software,Inc.)软件分析数据。缩脚阈值(PWT)数值越大，说明化合物的镇痛药效越强。
实验结果
大鼠SNL模型中的镇痛活性结果如表6和图6所示。
表6.本发明的化合物I C-1和加巴喷丁在大鼠SNL模型中给药后1小时缩脚阈值(PWT)统计数据
化合物 剂量 PWT(g),
对照组   3.967±0.775
化合物I C-1 10mg/kg 7.869±2.846
加巴喷丁 100mg/kg 6.352±1.897
从表6和图6的结果可知，与生理盐水对照组相比，本发明的化合物I C-1在10mg/kg的给药剂量下表现出非常强效的镇痛效果，具有显著性差异。与阳性对照组相比，在给药后1小时本发明化合物I C-1的镇痛活性与100mg/kg的加巴喷丁镇痛效果相当。大鼠SNL模型是经典的评价药物治疗神经疼痛的药效模型，因此本发明的化合物I C-1对神经疼痛具有优异的治疗效果。
实施例22
通过小鼠福尔马林疼痛模型考察实施例1制备的化合物I C-1对急性疼痛、炎症疼痛的治疗作用实验方法
取C57BL/6小鼠(雄性，9周)100只，小鼠每10只一组，随机分为10组,用于2个化合物在小鼠福尔马林疼痛模型的镇痛活性测试：分别为度洛西汀组和化合物I C-1组(实施例1制备的化合物)。实验开始前，让小鼠适应实验环境72h，期间无需禁食禁水。采用腹腔注射方式给予受试药物，剂量设置分别为：
度洛西汀组：空白Vehicle(空白生理盐水对照)、1mg/kg、5mg/kg、10mg/kg和20mg/kg；
化合物I C-1组：空白Vehicle(空白生理盐水对照，同度洛西汀空白组)、0.1mg/kg、0.5mg/kg、1mg/kg、5mg/kg和10mg/kg。
给药后将小鼠置于透明、通气的有机玻璃圆筒内，1h后于各组小鼠左后足底以微量进样器注射4％福尔马林溶液20μl，以微型摄像机实时记录小鼠的足部疼痛反应。以舔左足的时间长度作为疼痛反应的指标，分别观察记录0-10min(I相)和10-60min(II相)两时段内的舔足时间，进行统计学分析，并计算3个化合物的半数有效剂量(ED 50)：ED 50指的是与空白对照组相比添足时间降低一半的药物剂量。ED 50数值越小，表示化合物的镇痛有效剂量越低，其镇痛效果越好。
实验结果
化合物Ic-1和度洛西汀在小鼠福尔马林模型中不同给药剂量下(10-60min)期添足时间统计结果如表7和图7所示：
表.7化合物Ic-1和度洛西汀在小鼠福尔马林模型中不同给药剂量下在II相(10-60min)期添足时间统计结果
Figure PCTCN2021076337-appb-000067
Figure PCTCN2021076337-appb-000068
从表7和图7中可以看出，本发明的化合物Ic-1在1mg/kg的给药剂量下，其II相(10-60min)添足时间相较于空白Vehicle已经降低超过50％，在II相疼痛的镇痛药效ED 50为2.22mg/kg，而度洛西汀在II相疼痛的ED 50为8.00mg/kg。在同样的给药剂量下，本发明的化合物Ic-1的镇痛活性均显著优于度洛西汀。由以上数据可知，本发明的化合物Ic-1在小鼠福尔马林疼痛模型中表现出极强的镇痛活性。小鼠福尔马林模型是经典的评价药物急性疼痛和炎症疼痛药效的模型，因此本发明的化合物Ic-1对急性疼痛和炎症疼痛具有优异的治疗效果。
实施例23
实施例1制备的式I C-1化合物的盐晶型
XRPD：X射线粉末衍射；DSC：差示扫描量热分析；TGA：热重分析；DVS：动态水分吸附；X射线粉末衍射分析方法为：PANalytical X射线粉末衍射分析仪，工作电压：45kV，工作电流：40mA，使用Cu靶获得X射线粉末衍射图。
差示扫描量热(DSC)分析：仪器为TA Q2000/Discovery DSC2500；扫描速度：10℃/min；保护气体，氮气。
热重分析(TGA)分析：仪器为TA Q5000/Discovery TGA5500；扫描速度：10℃/min；保护气体，氮气。
动态水分吸附(DVS)分析：仪器为SMS公司(Surface Measurement Systems)生产的DVS Intrinsic；温度，25℃；载气，流速：氮气，200毫升/分钟；单位时间质量变换：0.002％/分钟；相对湿度范围：0％RH-95％RH。
核磁分析：仪器为Bruker 400M核磁共振仪。
高效液相色谱纯度(HPLC)在Agilent 1260高效液相色谱仪上采集。
离子色谱(IC)测试反荷离子确定摩尔比由Thermo ICS1100采集。
实施例1制备的式I C-1化合物为油状物，为了得到式I C-1化合物的固体形式，克服油状物的I C-1化合物造成的制药加工困难，本发明人通过悬浮搅拌的方法测试了盐酸、硫酸、L-天门冬氨酸、马来酸、L-焦谷氨酸、磷酸、L-谷氨酸、粘酸、L-(+)-酒石酸、富马酸、柠檬酸、D-葡萄糖醛酸、L-(-)-苹果酸、马尿酸、D-葡萄糖酸、羟基乙酸、乳酸、L-抗坏血酸、琥珀酸、己二酸、月桂酸、甲磺酸、草酸、丙二酸、苯甲酸、S-(+)-扁桃酸与式Ic-1化合物的成盐，结果表明，大多数酸配体形成的盐为油状物或者胶状物，而盐酸、马来酸、草酸、粘酸、富马酸和D-葡萄糖醛酸形成了固体晶体，筛选所得的这六种结晶盐通过TGA、DSC、HPLC、IC或者 1H NMR进行了进一步的表征。
式Ic-1化合物游离碱表征：
式Ic-1化合物游离碱用作起始原料以获得其他盐的形式。式Ic-1化合物游离碱为油状物，测试了室温条件下，式Ic-1化合物游离碱在11种常用溶剂中的溶解度。实验中，称取约2毫克的Ic-1化合物油状样品于3-毫升小瓶中，再逐步(50、50、200、700微升)加入相应溶剂后振荡直至溶液澄清。若溶剂加至1毫升后样品仍未溶清，则不再增加溶剂。根据固体样品质量、添加溶剂的体积和观察到的溶解现象计算得的粗略溶解度范围如表8所示，该数据为筛选实验中溶剂的选择提供参考。
表8 游离的Ic-1化合物在不同溶剂中的溶解度
溶剂 溶解度(mg/mL) 溶剂(v:v) 溶解度(mg/mL)
水 S<2.0 乙酸乙酯 S>36.0
正庚烷 7.6<S<23.0 四氢呋喃 S>40.0
甲醇 S>38.0 二甲基亚砜 S>38.0
乙醇 S>42.0 甲基异丁基酮 S>42.0
丙酮 S>46.0 甲醇:水(1:1) S>38.0
乙腈 S>54.0    
式Ic-1化合物的盐酸盐晶型A的制备方法：
称取292克式Ic-1化合物的游离碱，加入到4.4升乙酸乙酯溶液中，搅拌，用冰水浴降温至5～15℃，缓慢滴加37％的浓盐酸，调节体系pH至7，搅拌反应5分钟，有固体析出，过滤，滤饼用乙酸乙酯洗涤，将滤饼放入烘箱中(40～45℃)干燥至恒重，得到盐酸盐晶体193克，收率58.88％。
本实施例得到的式Ic-1化合物的盐酸盐晶型A的X射线粉末衍射数据如表9所示，XRPD图如图8，TGA/DSC叠加图如图9。
表9
2θ d值 相对强度％
10.09 8.76 52.42
11.25 7.87 12.51
16.85 5.26 75.73
18.27 4.86 100.00
21.27 4.18 82.46
21.84 4.07 31.52
22.20 4.00 71.08
22.89 3.88 83.32
23.86 3.73 57.43
25.40 3.51 39.30
26.76 3.33 25.94
28.18 3.17 36.31
28.75 3.11 25.68
32.57 2.75 10.18
从图9中可以看出，差示扫描量热分析图显示式Ic-1化合物的盐酸盐晶型A在加热至141.8℃附近开始出现吸热峰，热重分析图显示晶型A在加热至120℃时具有约1.1％的重量损失。
盐酸盐晶型A的变温XRPD测试结果如图10。对盐酸盐晶型A进行氮气吹扫以及氮气保护下加热至高温并降温至30℃后未观察到晶型变化，表明盐酸盐晶型A为无水晶型。进一步HPLC/IC结果显示，游离碱/酸的摩尔比为1:1。
式Ic-1化合物的马来酸盐晶型B的制备方法：
1)称取298.7毫克Ic-1化合物的游离碱和124.7毫克的马来酸至20毫升的玻璃小瓶中。
2)加入15毫升的乙酸乙酯形成悬浊液，在室温下置于磁力搅拌器上以750转/分钟的速率搅拌6天。
3)将悬浊液抽滤，室温真空干燥收集得固体约224.2毫克，收率为53.0％。
本实施例得到的式Ic-1化合物的马来酸盐晶型B的X射线粉末衍射数据如表10所示，XRPD图如图11，TGA/DSC叠加图如图12。
表10
2θ d值 相对强度％
9.70 9.12 15.51
11.83 7.48 18.79
15.22 5.82 10.21
16.30 5.44 13.25
18.23 4.87 14.69
18.82 4.72 11.41
19.23 4.61 72.02
19.56 4.54 37.68
21.15 4.20 8.57
21.83 4.07 1.84
23.60 3.77 12.70
24.04 3.70 100.00
24.70 3.60 42.79
24.93 3.57 15.64
26.35 3.38 12.47
28.15 3.17 24.14
28.94 3.09 9.61
32.59 2.75 3.74
33.31 2.69 3.29
34.74 2.58 2.77
35.94 2.50 1.13
38.18 2.36 1.62
从图12中可以看出，差示扫描量热分析图显示式Ic-1化合物的马来酸盐晶型B在加热至105.8℃附近开始出现吸热峰，热重分析图显示晶型B在加热至80℃时具有约0.8％的重量损失。式Ic-1化合物的马来酸盐晶型B的 1H NMR图谱如图13所示，结果显示，游离碱/酸的摩尔比为1:1。马来酸盐晶型B的变温XRPD测试结果如图14。对马来酸盐晶型B进行氮气吹扫以及氮气保护下加热至80℃后未观察到晶型变化，表明马来酸盐晶型B为无水晶型。
式Ic-1化合物的草酸盐晶型C的制备方法：
1)称取300.2毫克式Ic-1化合物的游离碱和94.2毫克的草酸至20毫升的玻璃小瓶中。
2)加入15毫升乙酸乙酯形成悬浊液，在室温下置于磁力搅拌器上以750转/分钟的速率搅拌6天。
3)将悬浊液抽滤，室温真空干燥后收集得到固体约309.5毫克，收率为78.5％。
本实施例得到的Ic-1化合物的草酸盐晶型C的X射线粉末衍射数据如表11所示，XRPD图如图15，TGA/DSC叠加图如图16。
表11
2θ d值 相对强度％
14.64 6.05 24.38
15.27 5.80 10.02
16.07 5.52 11.65
16.36 5.42 19.89
17.63 5.03 2.55
19.52 4.55 10.17
20.90 4.25 12.83
22.05 4.03 100.00
23.43 3.80 16.77
25.61 3.48 23.51
从图16中可以看出，差示扫描量热分析图显示式Ic-1化合物的晶型C在加热至152.2℃附近开始出现吸热峰，热重分析图显示晶型C在加热至100℃时具有约1.0％的重量损失
HPLC/IC结果显示，该样品中游离碱/酸的摩尔比为1:1。草酸盐晶型C的变温XRPD测试结果如图17。对草酸盐晶型C进行氮气吹扫以及氮气保护下加热至100℃后未观察到晶型变化。
式Ic-1化合物的粘酸盐晶型D的制备方法：
1).称取约7.4mg粘酸于HPLC小瓶中，加入0.5mL式Ic-1化合物的游离态样品的EtOAc储备液(40mg/mL)后，于50℃温度下磁力搅拌2天，析出固体后，离心分离，室温真空干燥，得到式Ic-1化合物的粘酸盐晶型D。
本实施例得到的式Ic-1化合物的粘酸盐晶型D的X射线粉末衍射数据如表12所示，XRPD图如图18，TGA/DSC叠加图如图19。
表12
2θ d值 相对强度％
3.79 23.30 40.31
11.28 7.84 100.00
14.21 6.23 3.82
15.81 5.61 39.14
16.97 5.23 14.93
17.71 5.01 8.72
19.48 4.56 87.36
20.98 4.23 18.69
23.91 3.72 21.51
25.88 3.44 10.60
27.16 3.28 3.35
28.40 3.14 10.06
29.49 3.03 5.70
30.74 2.91 6.11
32.33 2.77 3.19
34.50 2.60 3.87
35.42 2.53 3.41
36.16 2.48 2.23
从图19中可以看出，差示扫描量热分析图显示式Ic-1化合物的晶型D在加热至140.9℃附近开始出现吸热峰，热重分析图显示晶型D在加热至100℃时具有约1.45％的重量损失。
式Ic-1化合物的粘酸盐晶型D的 1H NMR图谱如图20所示，结果显示，，该样品中游离态与粘酸的摩尔比为2:1。
式Ic-1化合物的富马酸盐晶型E的制备方法：
1.称取约8.3mg富马酸于HPLC小瓶中，加入0.5mL式Ic-1化合物的游离态样品的 EtOAc储备液(40mg/mL)后，室温磁力搅拌约4天。
2.离心分离固体后室温真空干燥。
本实施例得到的晶型E的X射线粉末衍射数据如表13所示，XRPD图如图21，TGA/DSC叠加图如图22。
表13
2θ d值 相对强度％
4.70 18.82 9.14
9.41 9.40 5.61
13.34 6.64 7.80
14.17 6.25 28.48
15.63 5.67 6.40
17.62 5.03 2.92
18.95 4.68 44.00
22.75 3.91 10.62
23.76 3.74 100.00
25.66 3.47 5.88
26.93 3.31 14.03
28.69 3.11 6.48
31.34 2.85 2.76
从图22中可以看出，差示扫描量热分析图显示式Ic-1化合物的晶型E在加热至76.5℃附近开始出现吸热峰，热重分析图显示晶型E在加热至80℃时具有约2.17％的重量损失。
式Ic-1化合物的富马酸盐晶型E的 1H NMR图谱如图23所示，结果显示，该样品中游离态与富马酸的摩尔比为1:1。
式Ic-1化合物的D-葡萄糖醛酸盐晶型F的制备方法：
称取约13.5mg D-葡萄糖醛酸于HPLC小瓶中，加入0.5mL游离态Ic-1化合物的乙腈储备液(40mg/mL)后，室温磁力搅拌约4天,离心分离固体后室温真空干燥。
本实施例得到的晶型F的X射线粉末衍射数据如表14所示，XRPD图如图24，TGA/DSC叠加图如图25。
表14
2θ d值 相对强度％
4.77 18.52 100.00
8.34 10.60 54.57
10.87 8.14 46.79
16.13 5.49 97.98
17.54 5.06 55.33
19.53 4.55 74.17
20.06 4.43 60.75
21.25 4.18 44.80
23.42 3.80 29.68
25.93 3.44 31.65
从图25中可以看出，差示扫描量热分析图显示式Ic-1化合物的晶型F在加热至119.1℃附近开始出现吸热峰，热重分析图显示晶型F在加热至100℃时具有约2.71％的重量损失。
实施例24
本实施例考察实施例23制备的式Ic-1化合物的盐晶型的特性。
式Ic-1化合物的盐酸盐晶型A和马来酸盐晶型B的引湿性实验：
采用动态水分吸附(DVS)仪测试其引湿性，结果如图26和图28显示，盐酸盐晶型A和马来酸晶型B在25℃/80RH条件下重量变化小于0.2％，表明其均无引湿性，说明盐型易于存储，不需要置于干燥条件保存(参考中国药典2015年版(药物引湿性试验指导原则))。进一步XPRD测试表明，结果如图27和图29显示，盐酸盐晶型A和马来酸盐晶型B在DVS测试前后未观察到晶型的变化，表明Ic-1化合物的盐酸盐晶型A和马来酸晶型B在高湿环境中稳定。
式Ic-1化合物的盐酸盐晶型A和马来酸盐晶型B的动态溶解度：
式Ic-1化合物盐酸盐晶型A和马来酸盐晶型B分别用pH1.9SGF(模拟胃液)、pH6.5FaSSIF(模拟空腹状态下人工肠液)、pH 5.0FeSSIF(模拟进食状态下人工肠液)和水配置成饱和溶液，测试37℃下的动态溶解度。实验中，盐酸盐晶型A在H 2O、SGF、FaSSIF中的起始投料量为约20mg/mL，在FeSSIF中的起始投料量为约40mg/mL。马来酸盐晶型B在H 2O、SGF、FaSSIF和FeSSIF中的起始投料量为约10mg/mL。将样品密封固定在转速为25转/分钟的旋转盘上，并将旋转盘放置于37℃恒温箱中。分别在平衡1、4和24小时时间点取样，分离滤液测试HPLC浓度。结果如表15所示。
表15
Figure PCTCN2021076337-appb-000069
可以看出，式Ic-1化合物的盐酸盐晶型A和马来酸盐晶型B具有优异的溶解度，且在水中溶解度同时明显高于游离形式的Ic-1化合物，有利于提升药物的吸收和成药性。
式Ic-1化合物的盐酸盐晶型A和马来酸盐晶型B的固态稳定性研究：
分别称取约10毫克式Ic-1化合物的盐酸盐晶型A和式Ic-1化合物的马来酸晶型B加入HPLC小瓶中，瓶口用封口膜封口，膜上扎10个小孔，将小瓶置于25℃/60％RH和40℃/75％RH环境中放置4周，分别于第1周和第4周取样，对样品的纯度(采取HPLC分析)和晶型(采用X射线粉末衍射分析)进行考察。两种盐型的晶体放置1周和4周后，HPLC(高效液相色谱法)纯度无明显下降且未观察到晶型的变化，物理化学稳定性良好。稳定性样品放置前后的XRPD对比图如图30至图31所示。
表16
Figure PCTCN2021076337-appb-000070
式Ic-1化合物的盐酸盐晶型A水中平衡溶解度测试
在室温条件下对式Ic-1化合物的盐酸盐晶型A进行了水中24小时平衡溶解度测试，以～300mg/mL固体投料量投料，在水中磁力搅拌(750rpm)24小时后得到悬浊液，离心分离后将上清液经0.22微米PTFE滤膜过滤后进行HPLC测试，所得固体进行XRPD测试。结果显示，平衡溶解度测试后固体晶型未发生变化。盐酸盐晶型A在水中溶解度为210.5mg/mL(游离态浓度，折算为盐酸盐的浓度为237.3mg/mL)。
实施例25
本实施例考察实施例23制备的式Ic-1化合物的盐酸盐晶型A的绝对构型。
检测仪器：D8 Venture
仪器型号：D8 Venture
仪器参数：
光源：Mo靶                         X射线：Mo-Kɑ
Figure PCTCN2021076337-appb-000071
探测器：CMOS面探测器               分辨率：
Figure PCTCN2021076337-appb-000072
电流电压：50kV，1.4A               曝光时间：3s
面探测器至样品距离：40mm           测试温度：173(2)K
结构解析与精修过程：
采用SAINT程序对衍射数据进行积分还原后，采用SADABS程序对数据进行经验吸收校正；采用SHELXT2014通过直接法解析单晶结构，并采用最小二乘法对结构进行精修，氢原子精修过程采取各向同性计算处理获得，N上氢原子通过残余电子密度获得，C-H上氢原子通过计算加氢获得，并采取骑式模型对其精修处理。Flack常数为-0.09(5)，实施例23制备的式Ic-1化合物的盐酸盐晶型A的手性中心C4可以确认为S构型(如图32所示)。
实施例26
手动膜片钳检测实施例1制备的式I C-1化合物对TRPM8、TRPV1和TRPV4通道的作用
(1)细胞培养和处理
TRPM8细胞培养于直径35mm的细胞培养皿中，置于37℃，5％CO2的培养箱培养，每48小时按1:5比例进行传代，培养基配方：90％DMEM(Invitrogen)，10％胎牛血清(Gibco)，2mM L-glutamine，50μg/mL Hygromycin B(Invitrogen)和5μg/mL Blasticidine HCl(Invitrogen)。电生理膜片钳试验前，将细胞培液替换成含1μg/mL Doxycycline的培养基进行培养至少16小时以诱导表达TRPM8。
TRPV1细胞培养于直径35mm的细胞培养皿中，置于37℃，5％CO2的培养箱培养，每48小时按1:5比例进行传代，培养基配方：90％DMEM(Invitrogen)，10％胎牛血清(Gibco)，2mM L-glutamine，50μg/mL Hygromycin B(Invitrogen)和5μg/mL Blasticidine HCl(Invitrogen)。电生理膜片钳试验前，将细胞培液替换成含1μg/mL Doxycycline的培养基进行培养至少16小时以诱导表达TRPV1。
TRPV4细胞培养于直径35mm的细胞培养皿中，置于37℃，5％CO2的培养箱培养，每48小时按1:5比例进行传代，培养基配方：90％DMEM(Invitrogen)，10％胎牛血清(Gibco)，2mM L-glutamine，50μg/mL Hygromycin B(Invitrogen)和5μg/mL Blasticidine HCl(Invitrogen)。电生理膜片钳试验前，将细胞培液替换成含1μg/mL Doxycycline的培养基进行培养至少16小时以诱导表达TRPV4。
试验当天，吸走细胞培养液，用细胞外液淋洗一遍后加入0.25％Trypsin-EDTA (Invitrogen)溶液，在室温下消化1-2分钟。吸走消化液，用细胞外液重悬后将细胞转移到用于电生理记录的实验皿中备用。
(2)化合物准备
Menthol准备：取100.00mM Menthol DMSO母液加入到细胞外液中，1000倍稀释成100.00μM Menthol溶液。
AMTB准备：取20.00mM AMTB DMSO母液5μL加入到4995μL的100.00μM Menthol溶液中，1000倍稀释得到需要测试的最终浓度20.00μM AMTB。
Capsaicin准备：取1.00mM Capsaicin DMSO母液加入到细胞外液中，1000倍稀释成1.00μM Capsaicin溶液。
Capsazepine准备：取10.00mM Capsazepine DMSO母液5μL加入到4995μL的1.00μM Capsaicin溶液中，1000倍稀释得到需要测试的最终浓度10.00μM Capsazepine溶液。
GSK1016790A准备：取30.00μM GSK1016790A DMSO母液加入到细胞外液中，1000倍稀释成30.00nM GSK1016790A溶液。
RR(Ruthenium Red)准备：取10.00mM RR ddH2O母液20μL加入到4980μL的30.00nM GSK1016790A溶液中，250倍稀释得到需要测试的最终浓度40.00μM RR溶液。
Ic-1准备：取20.00mM或10.00mM Ic-1母液加入到相应通道的激动剂溶液中，1000或2000倍稀释成需要测试的最终浓度10.00M LDS溶液。
最终测试浓度中DMSO的含量不超过0.2％，此浓度的DMSO对通道电流没有影响。
(3)电生理记录过程
稳定表达TRP通道的HEK293细胞，在室温下用全细胞膜片钳技术记录相应激动剂诱发的通道电流。玻璃微电极由玻璃电极毛胚(BF150-86-10，Sutter)经拉制仪(P97，Sutter)拉制而成，灌注电极内液后的尖端电阻为2-5MΩ左右，将玻璃微电极插入放大器探头即可连接至膜片钳放大器。钳制电压和数据记录由pClamp 11软件通过电脑控制和记录，采样频率为10kHz，滤波频率为2kHz。在得到全细胞记录后，细胞钳制在0mV，300ms斜坡电压从-100mV至+100mV，每2s施加此电压刺激并给予激动剂诱发通道电流，电流诱发40s以上并确定无衰减后开始给药过程。每个测试浓度至少给予20s，每个浓度至少测试3个细胞(n≥3)。
(4)数据处理
数据分析处理采用pClamp 11，GraphPad Prism 5和Excel软件。不同化合物浓度对TRP通道电流(+100mV时诱发的电流幅度)的抑制程度用以下公式计算：
Inhibition％＝[1–(I/Io)]×100％
其中，Inhibition％代表化合物对电流的抑制百分率，I和Io分别表示在加药后和加药前电流的幅度。
(5)测试结果
在稳定表达TRPM8、TRPV1和TRPV4通道的HEK293细胞上，式I C-1化合物在10.00μM浓度下对三个通道电流的抑制百分率分别为41.07％、22.48％和4.68％(抑制百分率小于50％表明化合物对TRPM8的IC50大于10μM)，对应的阳性化合物AMTB、Capsazepine和RR的抑制百分率都为90％以上。由以上结果可知化合物Ic-1是TRPA1特异性的抑制剂。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求 书所限定的范上围。

Claims (15)

  1. 一种式I化合物的制备方法，其特征在于，所述的方法包括步骤：
    (1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式a化合物与式b化合物反应，得到式I化合物；`
    Figure PCTCN2021076337-appb-100001
    其中：
    环A为取代或未取代的4-12元碳环、取代或未取代的4-12元杂环、取代或未取代的5-12元杂芳环、取代或未取代的C 6-C 12芳环；
    R 1和R 2各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
    X和Y各自独立地为碳原子、氧原子、硫原子或氮原子；
    R 3为氢、卤素、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
    R 4为卤素；
    R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基，取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
    W为O或S；
    n为1、2或3；
    其中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、C 6-C 12芳基、5-10元杂芳基；
    其中，所述的杂环、杂芳环和杂芳基各自独立地具有1-3个(优选为1、2或3个)选自N、O和S杂原子。
  2. 一种式I-1化合物的制备方法，其特征在于，所述的方法包括步骤：
    (1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式a化合物与式b化合物反应，得到式I化合物；
    Figure PCTCN2021076337-appb-100002
    (2)在第二溶剂中，在第二碱试剂的存在下，式I化合物与式c所示的N-去甲基化试剂反应，得到式I-a化合物；
    Figure PCTCN2021076337-appb-100003
    (3)在第三溶剂中，在第三碱试剂的存在下，式I-a化合物发生水解反应，得到式I-1化合物；
    Figure PCTCN2021076337-appb-100004
    其中，
    环A为取代或未取代的4-12元碳环、取代或未取代的4-12元杂环、取代或未取代的5-12元杂芳环、取代或未取代的C 6-C 12芳环；
    R 1和R 2各自独立为取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 7环烷基；
    X和Y各自独立地为碳原子、氧原子、硫原子或氮原子；
    R 3为氢、卤素、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
    R 4为卤素；
    R 5、R 6、R 7、R 8、R 9、R 10和R 11各自独立为氢、取代或未取代的C 1-C 12烷基、取代或未取代的C 3-C 12环烷基，取代或未取代的5-12元杂芳基、取代或未取代的C 6-C 12芳基；
    R 12为取代或未取代的C 1-C 6烷基、取代或未取代的C 3-C 12环烷基、取代或未取代的C 6-C 16芳基、取代或未取代的5-16元杂芳基、取代或未取代的C 1-C 6烷基-W-、取代或未取代的C 3-C 12环烷基-W-、取代或未取代的C 6-C 16芳基-W-、取代或未取代的5-16元杂芳基-W-；
    R 13为卤素；
    W为O或S；
    n为1、2或3；
    其中，所述的任一“取代”是指基团上的1-4个(优选为1、2、3或4个)氢原子各自独立地被选自下组的取代基所取代：C 1-C 6烷基、C 3-C 7环烷基、C 1-C 3卤代烷基、卤素、硝基、氰基、氨基、羟基、＝O、C 1-C 4羧基、C 2-C 4酯基、C 2-C 4酰胺基、C 1-C 6烷氧基、C 1-C 6卤代烷氧基、苄基、C 6-C 12芳基、5-10元杂芳基；
    其中，所述的杂环、杂芳环和杂芳基各自独立地具有1-3个(优选为1、2或3个)选自N、O和S杂原子。
  3. 如权利要求2所述的方法，其特征在于，所述的方法包括步骤：
    (1)在第一溶剂中，在第一催化剂和第一碱试剂的存在下，式i化合物与式ii化合物反应，得到式Ic-8化合物；
    Figure PCTCN2021076337-appb-100005
    (2)在第二溶剂中，在第二碱试剂的存在下，式Ic-化合物与氯甲酸苯酯反应，得到式iii化合物；
    Figure PCTCN2021076337-appb-100006
    (3)在第三溶剂中，在第三碱试剂的存在下，式iii化合物发生水解反应，得到式Ic-1化合物；
    Figure PCTCN2021076337-appb-100007
  4. 一种式Ic-1化合物的盐酸盐或盐酸盐晶型A，
    Figure PCTCN2021076337-appb-100008
    所述的盐酸盐晶型A的X-射线粉末衍射图谱在2θ角为18.27±0.2°、21.27±0.2°、22.89±0.2°处具有特征峰。
  5. 如权利要求4所述的Ic-1化合物的盐酸盐或盐酸盐晶型A，其特征在于，
    所述盐酸盐晶型A在1个或多个选自下组的2θ值处具有特征峰：10.09±0.2°、11.25±0.2°、16.85±0.2°、18.27±0.2°、21.27±0.2°、21.84±0.2°、22.20±0.2°、22.89±0.2°、23.86±0.2°、25.40±0.2°、26.76±0.2°、28.18±0.2°、28.75±0.2°、32.57±0.2°。
  6. 如权利要求4所述的Ic-1化合物的盐酸盐或盐酸盐晶型A，其特征在于，
    所述的盐酸盐晶型A的X-射线粉末衍射图谱具有一个或选自下组的2θ值处的特征峰和峰强度：
    2θ值 d值 相对强度％ 10.09 8.76 52.42 11.25 7.87 12.51 16.85 5.26 75.73 18.27 4.86 100.00 21.27 4.18 82.46 21.84 4.07 31.52 22.20 4.00 71.08 22.89 3.88 83.32 23.86 3.73 57.43 25.40 3.51 39.30 26.76 3.33 25.94 28.18 3.17 36.31 28.75 3.11 25.68 32.57 2.75 10.18。
  7. 如权利要求4所述的Ic-1化合物的盐酸盐或盐酸盐晶型A，其特征在于，
    所述盐酸盐晶型A具有基本如图8所示的X-射线粉末衍射特征峰。
  8. 如权利要求4所述的Ic-1化合物的盐酸盐或盐酸盐晶型A，其特征在于，所述的Ic-1化合物的盐酸盐晶型A具有选自下组的一种或多种特征峰：
    所述盐酸盐晶型A的差示扫描量热分析(DSC)图在加热至141.8±5℃(较佳地±4℃、±3℃、±2℃或±1℃)开始出现吸热峰；
    所述盐酸盐晶型A的差示扫描量热分析(DSC)图基本如图9所示；
    所述盐酸盐晶型A的热重分析(TGA)图在加热至120℃时具有约1.1±0.5％(较佳地±0.4％、±0.3％、0.2％或0.1％)的重量损失；和/或
    所述盐酸盐晶型A的热重分析(TGA)图基本如图9所示。
  9. 一种制备如权利要求4所述的Ic-1化合物的盐酸盐晶型A的方法，其特征在于，所述的方法包括步骤：
    (a)式Ic-1化合物与第一有机溶剂混合后，在5～15℃下滴加盐酸，调节体系pH为6-8，反应析出固体，过滤得到式Ic-1化合物的盐酸盐晶型A；
    所述的第一有机溶剂包括乙酸乙酯。
  10. 一种式Ic-1化合物的马来酸盐或马来酸盐晶型B，
    Figure PCTCN2021076337-appb-100009
    所述的马来酸盐晶型B的X-射线粉末衍射图谱在2θ角为19.23±0.2°、24.04±0.2°、24.70±0.2°处具有特征峰。
  11. 一种式Ic-1化合物的草酸盐或草酸盐晶型C，
    Figure PCTCN2021076337-appb-100010
    所述的草酸盐晶型C的X-射线粉末衍射图谱在2θ角为14.64±0.2°、22.05±0.2°、25.61±0.2°处具有特征峰。
  12. 一种式Ic-1化合物的粘酸盐或粘酸盐晶型D，
    Figure PCTCN2021076337-appb-100011
    所述的粘酸盐晶型D的X-射线粉末衍射图谱在2θ角为3.79±0.2°、11.28±0.2°、19.48±0.2°处具有特征峰。
  13. 一种式Ic-1化合物的富马酸盐或富马酸盐晶型E，
    Figure PCTCN2021076337-appb-100012
    所述的富马酸盐晶型E的X-射线粉末衍射图谱在2θ角为14.17±0.2°、18.95±0.2°、23.76±0.2°处具有特征峰。
  14. 一种式Ic-1化合物的D-葡萄糖醛酸盐或D-葡萄糖醛酸盐晶型F，
    Figure PCTCN2021076337-appb-100013
    所述的D-葡萄糖醛酸盐晶型F的X-射线粉末衍射图谱在2θ角为4.77±0.2°、16.13±0.2°、19.53±0.2°处具有特征峰。
  15. 一种如权利要求4所述的式Ic-1化合物的盐酸盐晶型A、如权利要求10所述的式Ic-1化合物的马来酸盐晶型B、如权利要求11所述的式Ic-1化合物的草酸盐晶型C、如权利要求12所述的式Ic-1化合物的粘酸盐晶型D、如权利要求13所述的式Ic-1化合物的富马酸盐晶型E、或如权利要求14所述的式Ic-1化合物的D-葡萄糖醛酸盐晶型F的用途，其特征在于，用于(a)制备瞬时受体电位通道蛋白(TRP)抑制剂；和/或(b)制备预防和/或治疗与瞬时受体电位通道蛋白(TRP)相关的疾病的药物。
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