P2X3 inhibitors and uses thereof

Abstract

The present application provides a new compound which effectively antagonizes P2X3 receptor, which is a compound shown in formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof, a preparation method thereof, and a use thereof in the preparation of a medicine.

Description

[0001] Priority information

[0002] This application claims priority and benefits to patent application No. 202011411777.9, filed with the China National Intellectual Property Administration on December 4, 2020, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention belongs to the field of pharmaceutical chemistry. Specifically, this invention relates to P2X3 inhibitors and their uses. More specifically, this invention relates to a benzamide compound and its preparation method, as well as its use in the preparation of drugs. Background Technology

[0004] The P2X receptor is a non-selective ATP-gated ion channel receptor, also known as a purinergic receptor, that binds to extracellular ATP, primarily derived from damaged or inflamed tissues. This receptor is widely expressed in the nervous, immune, cardiovascular, skeletal, gastrointestinal, respiratory, and endocrine systems, and participates in various physiological processes, including regulation of heart rate and contractility, regulation of vascular tone, regulation of nociception, especially chronic pain, vas deferens contraction during ejaculation, bladder contraction during urination, platelet aggregation, macrophage activation, apoptosis, and neuron-glial interactions. The aforementioned P2X receptors include seven homologous receptors: P2X1, P2X2, P2X3, P2X4, P2X5, P2X6, and P2X7, and three heterologous receptors: P2X2 / 3, P2X4 / 6, and P2X1 / 5.

[0005] P2X3 is a subtype of the P2X receptor family that is selectively expressed in dorsal root ganglia of nerve endings, spinal cord, and brain neurons, specifically in small to medium diameter primary sensory neurons.

[0006] Numerous studies have shown that activation of P2X3 and P2X2 / 3 expressed in primary sensory neurons plays a crucial role in acute injury, hyperalgesia, and hypersensitivity responses in rodents. Many studies have demonstrated that upregulation of P2X3 receptor expression can lead to hyperalgesia and participate in pain signaling. P2X3 knockout mice exhibit reduced pain responses, and P2X3 receptor antagonists have shown a role in reducing nociception in pain and inflammatory pain models.

[0007] P2X3 is distributed in primary afferent nerves surrounding the airways and can regulate cough. Studies have shown that ATP released from damaged or inflamed airway tissue acts on P2X3 receptors in primary neurons, triggering depolarization and action potentials. These potentials transmit cough impulses, initiating coughing. P2X3 receptors play an important role in the cough reflex hypersensitivity response; by antagonizing P2X3 receptor binding, the hypersensitivity response of the cough reflex can be inhibited, thereby suppressing excessive coughing in patients with chronic cough. Furthermore, research has shown that P2X3 antagonists can treat chronic obstructive pulmonary disease, pulmonary fibrosis, pulmonary hypertension, or asthma; therefore, P2X3 antagonists also hold promise as new drugs for treating these diseases.

[0008] P2X3 has been reported to be involved in the afferent pathway controlling the bladder capacity reflex; P2X3 knockout mice exhibit significantly reduced urination frequency and significantly increased bladder capacity. Therefore, inhibiting the binding of P2X3 receptor antagonists to the P2X3 receptor could be effective in treating conditions involving urinary storage and voiding disorders, such as overactive bladder. Thus, P2X3 antagonists may be potential drugs for treating overactive bladder and related diseases.

[0009] P2X3 antagonists show great promise. Currently, commonly used cough medications such as gabapentin, morphine, and amitriptyline, or treatments using speech pathology, can improve coughs in many patients, but they are not suitable for all patients. Moreover, centrally acting drugs such as gabapentin may produce adverse side effects and are not suitable for long-term use. There is an urgent clinical need to develop long-term medications for chronic refractory coughs to provide doctors with treatment options. Therefore, the development of P2X3 antagonists is of great clinical significance. Summary of the Invention

[0010] The present invention aims to provide a P2X3 receptor antagonist that can be used to prepare drugs for treating cough, pain, respiratory diseases and genitourinary diseases.

[0011] In a first aspect, the present invention provides a compound, which is a compound of Formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug:

[0012]

[0013] Where L represents non-existent,

[0014] R 1 Selected from hydrogen, halogen, hydroxyl, cyano, amino, unsubstituted or R-substituted a Substituted C1-C6 alkyl, unsubstituted or R a Substituted C1-C6 alkyl-O-, unsubstituted or R-substituted aSubstituted C3-C6 cycloalkyl, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups, or unsubstituted or R a Substituted 4-8 membered heterocyclic alkenyl groups; the R-substituted a Substituted C1-C6 alkyl groups, the R-substituted a Substituted C1-C6 alkyl-O-, the R a Substituted C3-C6 cycloalkyl groups, the R a The substituted 5-8 aryl group, the R- a Substituted 5-8 aryl groups, the aforementioned R a Substituted 4-8 membered heterocyclic alkyl groups, or, as described above, R a In the substituted 4-8 membered heterocyclic alkenyl groups, the R-substituted group a The substituents are each independently selected from one or more of the following substituents: halogen, hydroxyl, cyano, amino, oxo, C1-C6 alkyl, C1-C6 alkyl-O-, or, C1-C6 alkyl substituted with 1 to 5 identical or different halogens; when there are multiple substituents, the substituents are identical or different;

[0015] The unsubstituted or R a In the substituted 5-8 membered heteroaryl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted groups... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkenyl groups, the heteroatoms are selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3.

[0016] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a Substituted C1-C6 alkyl groups, or unsubstituted or R a When the C1-C6 alkyl group is substituted with -O-, the C1-C6 alkyl group is independently a C1-C4 alkyl group, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl or isobutyl.

[0017] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted C3-C6 cycloalkyl group is used, the C3-C6 cycloalkyl group is independently cyclopropane, cyclobutane, cyclopentane or cyclohexane, preferably cyclopropane or cyclobutane.

[0018] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted 5-8 aryl group is used, the 5-8 aryl group is independently phenyl or naphthyl, preferably phenyl.

[0019] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted 5-8 heteroaryl group is used, the 5-8 heteroaryl group is independently pyrrole, pyrazole, triazole, furan, oxazole, thiophene, thiazole, pyridine, pyrazine or pyrimidine, preferably pyrazole, furan, thiophene or pyridine.

[0020] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted 4-8 membered heterocyclic alkyl group is used, the 4-8 membered heterocyclic alkyl group is independently aziridine, aziridine propane, oxacyclobutane, tetrahydropyrrolidinyl, morpholinyl, piperazine, tetrahydrofuranyl, 2-oxa-6-azaspiro[3.3]hept-6-yl, 2-oxa-5-azabicyclo[2.2.1]hept-5-yl, hexahydropyran, or tetrahydro-2H-thiopyran 1,1-dioxide.

[0021] In a preferred embodiment of the present invention, when R 1 For not replaced or by R a When the substituted 4-8 membered heterocyclic alkenyl group is used, the 4-8 membered heterocyclic alkenyl group is independently dihydropyridyl, tetrahydropyridyl, tetrahydropyrimidinyl, pyrrolinyl, imidazolinyl, pyrazolinyl, dihydroimidazolinyl, dihydropyrazolyl, dihydrooxazolyl, dihydrooxadiazolyl, dihydrothiazolinyl, dihydroisothiazolinyl, dihydrothiophenyl, dihydropyrrolyl, 3,4-dihydro-2H-pyranyl, dihydrofuranyl, dihydropyrazinyl, dihydropyrimidinyl or fluorodihydrofuranyl, preferably 1,2,3,4-tetrahydropyridyl, 1,2-dihydropyridyl, 1,4-dihydropyridyl, 1,2,3,6-tetrahydropyridyl, 3,4-dihydro-2H-pyranyl or dihydrofuranyl.

[0022] In a preferred embodiment of the present invention, when R 1 When the substituted C1-C6 alkyl, substituted C1-C6 alkyl-O-, substituted C3-C6 cycloalkyl, substituted 5-8 aryl, substituted 5-8 heteroaryl, or substituted 4-8 heterocycloalkyl, the number of substituted ...

[0023] In a preferred embodiment of the present invention, when R 1The substituted C1-C6 alkyl, substituted C1-C6 alkyl-O-, substituted C3-C6 cycloalkyl, substituted 5-8 aryl, substituted 5-8 heteroaryl, or substituted 4-8 heterocycloalkyl, wherein each of the substituted substances is independently a halogen, and the halogen is F, Cl, Br or I, preferably F.

[0024] In a preferred embodiment of the present invention, when R 1 When the substituted C1-C6 alkyl, substituted C1-C6 alkyl-O-, substituted C3-C6 cycloalkyl, substituted 5-8 aryl, substituted 5-8 heteroaryl, or substituted 4-8 heterocycloalkyl, and each of the substituted alkyl groups is independently C1-C6 alkyl, the C1-C6 alkyl group in the substituted alkyl group is independently C1-C4 alkyl, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl.

[0025] In a preferred embodiment of the present invention, when R 1 When the substituted C1-C6 alkyl, substituted C1-C6 alkyl-O-, substituted C3-C6 cycloalkyl, substituted 5-8 aryl, substituted 5-8 heteroaryl, or substituted 4-8 heterocycloalkyl, the substituted substituted alkyl is independently oxo.

[0026] In a preferred embodiment of the present invention, when R 1 When the substituted C1-C6 alkyl, substituted C1-C6 alkyl-O-, substituted C3-C6 cycloalkyl, substituted 5-8 aryl, substituted 5-8 heteroaryl, or substituted 4-8 heterocycloalkyl, the substituted substance is independently a hydroxyl group.

[0027] In a preferred embodiment of the present invention, for The ring A is unsubstituted or replaced by R. a Substituted nitrogen-containing 4-8 membered heterocyclic alkyl groups, the unsubstituted or R a In the substituted nitrogen-containing 4-8 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3;

[0028] The L and R a It has the definition described above.

[0029] In a preferred embodiment of the present invention, R 1 -C(CH3)2OH,

[0030] In a preferred embodiment of the present invention, L represents non-existence, R 1 -C(CH3)2OH,

[0031] In a preferred embodiment of the present invention, the compound is a compound from Scheme 1, Scheme 2, or Scheme 3; wherein,

[0032] Option 1:

[0033] The compound has the structure shown in the following formula:

[0034] R 1 Selected from hydrogen, halogen, hydroxyl, cyano, amino, unsubstituted or R-substituted a Substituted C1-C6 alkyl, unsubstituted or R a Substituted C1-C6 alkyl-O-, unsubstituted or R-substituted a Substituted C3-C6 cycloalkyl, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups, or unsubstituted or R a Substituted 4-8 membered heterocyclic alkenyl groups; the R-substituted a Substituted C1-C6 alkyl groups, the R-substituted a Substituted C1-C6 alkyl-O-, the R a Substituted C3-C6 cycloalkyl groups, the R a The substituted 5-8 aryl group, the R- a Substituted 5-8 aryl groups, the aforementioned R a Substituted 4-8 membered heterocyclic alkyl groups, or, as described above, R a In the substituted 4-8 membered heterocyclic alkenyl groups, the R-substituted group a The substituents are each independently selected from one or more of the following substituents: halogen, hydroxyl, cyano, amino, oxo, C1-C6 alkyl, C1-C6 alkyl-O-, or, C1-C6 alkyl substituted with 1 to 5 identical or different halogens; when there are multiple substituents, the substituents are identical or different;

[0035] The unsubstituted or R a In the substituted 5-8 membered heteroaryl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted groups... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkenyl groups, the heteroatoms are selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3.

[0036] In a preferred embodiment of the present invention, in scheme 1, R 1 Selected from unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups, the unsubstituted or R a In the substituted 4-8 membered heterocyclic alkyl groups, each of the substitutions is independently selected from one or more of the following substituents: hydroxyl, C1-C6 alkyl or C1-C6 alkyl-O-, preferably, the 4-8 membered heterocyclic alkyl groups are independently aziridine, aziridine propane, oxacyclobutane, tetrahydropyrrolidinyl, morpholinyl, piperazine, tetrahydrofuranyl, 2-oxa-6-azaspiro[3.3]hept-6-yl, 2-oxa-5-azabicyclo[2.2.1]hept-5-yl, hexahydropyran or tetrahydro-2H-thiopyran 1,1-dioxide.

[0037] Option 2:

[0038] The compound has the structure shown in the following formula:

[0039] R 1 Selected from hydrogen, halogen, hydroxyl, cyano, amino, unsubstituted or R-substituted a Substituted C1-C6 alkyl, unsubstituted or R a Substituted C1-C6 alkyl-O-, unsubstituted or R-substituted a Substituted C3-C6 cycloalkyl, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups, or unsubstituted or R a Substituted 4-8 membered heterocyclic alkenyl groups; the R-substituted a Substituted C1-C6 alkyl groups, the R-substituted a Substituted C1-C6 alkyl-O-, the R a Substituted C3-C6 cycloalkyl groups, the R a The substituted 5-8 aryl group, the R- a Substituted 5-8 aryl groups, the aforementioned R a Substituted 4-8 membered heterocyclic alkyl groups, or, as described above, R a In the substituted 4-8 membered heterocyclic alkenyl groups, the R-substituted group a The substituents are each independently selected from one or more of the following substituents: halogen, hydroxyl, cyano, amino, oxo, C1-C6 alkyl, C1-C6 alkyl-O-, or, C1-C6 alkyl substituted with 1 to 5 identical or different halogens; when there are multiple substituents, the substituents are identical or different;

[0040] The unsubstituted or R a In the substituted 5-8 membered heteroaryl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted groups... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkenyl groups, the heteroatoms are selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3.

[0041] In a preferred embodiment of the present invention, R 1 For not replaced or by R a The substituted 4-8 membered heterocyclic alkyl group, said to be R a In the substituted 4-8 membered heterocyclic alkenyl groups, the R-substituted group a The substitutions are each independently selected from one or more of the following substituents: halogen, hydroxyl, cyano, amino, oxo, C1-C6 alkyl, C1-C6 alkyl-O-, or, C1-C6 alkyl substituted with 1 to 5 identical or different halogens; when there are multiple substituents, the substituents are identical or different; the unsubstituted or R-substituted substituents are... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3.

[0042] In a preferred embodiment of the present invention, in scheme 2, R 1 It is selected from 4-6 membered heterocyclic alkyl groups, the heteroatoms are selected from one or more of N, S, O and P, the number of heteroatoms is 1-3, and it is preferably aziridine propane, aziridine butane or tetrahydropyrrole alkyl.

[0043] Option 3:

[0044] The compound has the structure shown in the following formula:

[0045] R 1 Selected from hydrogen, halogen, hydroxyl, cyano, amino, unsubstituted or R-substituted a Substituted C1-C6 alkyl, unsubstituted or R a Substituted C1-C6 alkyl-O-, unsubstituted or R-substituted a Substituted C3-C6 cycloalkyl, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 5-8 aryl groups, unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups, or unsubstituted or R a Substituted 4-8 membered heterocyclic alkenyl groups; the R-substituteda Substituted C1-C6 alkyl groups, the R-substituted a Substituted C1-C6 alkyl-O-, the R a Substituted C3-C6 cycloalkyl groups, the R a The substituted 5-8 aryl group, the R- a Substituted 5-8 aryl groups, the aforementioned R a Substituted 4-8 membered heterocyclic alkyl groups, or, as described above, R a In the substituted 4-8 membered heterocyclic alkenyl groups, the R-substituted group a The substituents are each independently selected from one or more of the following substituents: halogen, hydroxyl, cyano, amino, oxo, C1-C6 alkyl, C1-C6 alkyl-O-, or, C1-C6 alkyl substituted with 1 to 5 identical or different halogens; when there are multiple substituents, the substituents are identical or different;

[0046] The unsubstituted or R a In the substituted 5-8 membered heteroaryl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted groups... a In the substituted 4-8 membered heterocyclic alkyl groups, the heteroatoms are selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the unsubstituted or R-substituted... a In the substituted 4-8 membered heterocyclic alkenyl groups, the heteroatoms are selected from one or more of N, S, O and P, and the number of heteroatoms is 1-3.

[0047] In a preferred embodiment of the present invention, R 1 For not replaced or by R a Substituted C1-C3 alkyl groups, said to be R a In the substituted C1-C3 alkyl group, the substitution is one or more hydroxyl substitutions.

[0048] In a preferred embodiment of the present invention, in scheme 3, R 1 Selected from unsubstituted or R a Substituted C2-C4 alkyl, or, unsubstituted or R a Substituted 4-6 membered heterocyclic alkyl groups, the unsubstituted or R a In the substituted C2-C4 alkyl group, the one being R a The substituent is selected from hydroxyl groups, and the unsubstituted or R-substituted group is selected from hydroxyl groups. a In the substituted 4-6 membered heterocyclic alkyl group, the heteroatom is selected from one or more of N, S and O, and the number of heteroatoms is 1 or 2, and the substituent is selected from oxo.

[0049] In a preferred embodiment of the present invention, for The ring A is unsubstituted or replaced by R. a The substituted nitrogen-containing 3- or 4-membered heterocyclic alkyl group, wherein the heteroatom is selected from one or more of N, S, and O, and the substituent is selected from one or more hydroxyl groups and one or more C1-C3 alkyl groups; and / or, ring A is unsubstituted or R-substituted. a The substituted nitrogen- and oxygen-containing 5- or 6-membered heterocyclic alkyl group, wherein the heteroatoms are selected from one or more of N, S and O, and the number is 1 to 3, and the substituents are selected from C1-C3 alkyl groups.

[0050] In a preferred embodiment of the present invention, ring A is selected from... Wherein at least one of X1, X2, X3, X4, and X5 is selected from O or S, the rest are selected from C or N, and X6 is selected from C1-C3 alkyl; preferably, the ring A is selected from X6 is selected from C1-C3 alkyl groups.

[0051] In a preferred embodiment of the present invention, ring A is a 6-membered spirobi-heterocyclic ring or a bridged bi-heterocyclic ring containing nitrogen and oxygen, and the number of heteroatoms is 1 to 3.

[0052] In a preferred embodiment of the present invention, the compound represented by Formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs are selected from any of the following compounds:

[0053]

[0054] In a second aspect, the present invention provides a pharmaceutical composition comprising the above-described compound, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and pharmaceutically acceptable pharmaceutical carriers, diluents or excipients.

[0055] According to specific embodiments of the present invention, the pharmaceutical compositions of the present invention, comprising a therapeutically effective dose of the aforementioned compounds, their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and pharmaceutically acceptable pharmaceutical carriers, diluents, or excipients, can be mixed to prepare a pharmaceutical formulation suitable for oral or parenteral administration. Administration methods include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulation can be administered via any route, such as by infusion or bolus, or by absorption through the epithelium or mucous membranes (e.g., oral mucosa or rectum). Administration can be systemic or local. Examples of orally administered formulations include solid or liquid dosage forms, specifically including tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The formulation can be prepared by methods known in the art and contains carriers, diluents, or excipients conventionally used in the field of pharmaceutical formulations.

[0056] In a third aspect, the present invention provides the use of the above-described compounds, their tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-described pharmaceutical compositions, in the preparation of a medicament for treating diseases related to P2X3.

[0057] According to specific embodiments of the present invention, the use of the above-described compound or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-described pharmaceutical compositions in the preparation of a medicament for the treatment or prevention of pain. These pains include, for example, chronic pain, acute pain, endometriosis pain, neuropathic pain, back pain, cancer pain, inflammatory pain, surgical pain, migraine, or visceral pain, preferably endometriosis pain and neuropathic pain.

[0058] According to specific embodiments of the present invention, the use of the above-described compound or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs or the above-described pharmaceutical compositions in the preparation of a medicament for the treatment or prevention of diseases of the urogenital system. These diseases include, for example, reduced bladder capacity, frequent urination, urge incontinence, stress incontinence, bladder hyperresponsiveness, benign prostatic hyperplasia, prostatitis, detrusor hyperreflexia, urinary frequency, nocturia, urinary urgency, overactive bladder, pelvic hypersensitivity, urethritis, pelvic pain syndrome, prostatodynia, cystitis, or idiopathic bladder hypersensitivity, preferably overactive bladder.

[0059] According to specific embodiments of the present invention, the use of the above-described compound or its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-described pharmaceutical compositions in the preparation of a medicament for the treatment or prevention of respiratory diseases. These diseases include, for example, chronic obstructive pulmonary disease, pulmonary hypertension, pulmonary fibrosis, asthma, obstructive sleep apnea, chronic cough, refractory chronic cough, and acute cough.

[0060] Terms and Definitions

[0061] Unless otherwise stated, the terms and definitions used in this application, including those set forth in the specification and claims, are as follows.

[0062] Those skilled in the art will understand that, according to the conventions used in the art, in the structural formula of this application, Used to describe chemical bonds, which are points where a portion or a substituent is connected to a core or skeletal structure.

[0063] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0064] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable, non-toxic salt of an acid or base, including salts of inorganic acids and bases, and salts of organic acids and bases.

[0065] The term "pharmaceutical composition" refers to a mixture of one or more compounds described herein, or physiologically / pharmaceutical acceptable salts or prodrugs thereof, with other chemical components, such as physiologically / pharmaceutical acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compound to a living organism.

[0066] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of the term "excipient" include, without limitation, binders, disintegrants, lubricants, flow aids, stabilizers, fillers, and diluents. The term "solvent" refers to a compound of the present invention or a salt thereof comprising a stoichiometric or non-stoichiometric solvent bound by intermolecular non-covalent forces; when the solvent is water, it is a hydrate.

[0067] The term "prodrug" refers to a compound of the present invention that can be converted into a biologically active form under physiological conditions or by solvation. The prodrugs of the present invention are prepared by modifying functional groups in the compound; this modification can be performed conventionally or removed in vivo to obtain the parent compound. Prodrugs comprise compounds formed by attaching a hydroxyl or amino group to any group within the compound of the present invention. When a prodrug of the compound of the present invention is administered to a mammalian individual, the prodrug is cleaved to form a free hydroxyl group and a free amino group.

[0068] The term "stereoisomer" refers to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, non-corresponding isomers, and conformational isomers.

[0069] Depending on the choice of raw materials and methods, the compounds of the present invention may exist as one or a mixture of possible isomers, for example as purely optical isomers, or as mixtures of isomers, such as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or multiple chiral centers) in the molecule. The prefixes D and L or (+) and (–) are symbols used to specify the plane-polarized rotation of light induced by the compound, where (–) or L indicates that the compound is levorotatory. Compounds with the prefix (+) or D are dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be called enantiomers, and mixtures of said isomers are generally referred to as mixtures of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or method. Many geometric isomers of alkenes, C=N double bonds, etc., can also exist in the compounds described herein, and all such stable isomers are considered in this invention. When the compounds described herein contain an alkene double bond, unless otherwise stated, such double bond includes E and Z geometric isomers. If the compound contains a disubstituted cycloalkyl group, the substituent of the cycloalkyl group may be in cis or trans (cis- or trans-) configuration.

[0070] When the bonds of the chiral carbon in the formulas of this invention are depicted as straight lines, it should be understood that both the (R) and (S) configurations of the chiral carbon and the resulting enantiomerically pure compounds and mixtures thereof are included within the scope of the general formula. The illustration of racemic or enantiomerically pure compounds in this document is derived from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise stated, wedge-shaped and dashed bonds represent the absolute configuration of a stereocenter.

[0071] Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral formulations, or resolved using conventional techniques. Compounds of the present invention containing asymmetrically substituted carbon atoms can be separated in either an optically active or racemic form. Resolution of racemic mixtures of compounds can be performed by any of many methods known in the art. Exemplary methods include fractional recrystallization using a chiral resolving acid, which is an optically active salt-forming organic acid. Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids such as the D and L forms of β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization methods include stereoisomerically pure forms of α-methylbenzylamine (e.g., S and R forms or diastereoisomerically pure forms), 2-phenylglycine, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, etc. Resolution of racemic mixtures can also be performed by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). High-performance liquid chromatography (HPLC) or supercritical fluid chromatography (SFC) can be used. The specific method, elution conditions, and column selection can be chosen by those skilled in the art based on the structure of the compound and experimental results. Furthermore, any enantiomer or diastereomeric form of the compound described in this invention can be obtained through stereoorganic synthesis using optically pure starting materials or reagents with known configurations.

[0072] The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom between two positions within a molecule. The compounds of this invention can exhibit tautomerism. Tautomers can exist in two or more interconvertible forms. Proton-transfer tautomers arise from the migration of covalently bonded hydrogen atoms between two atoms. Tautomers generally exist in equilibrium form; attempts to isolate a single tautomer typically yield a mixture whose physicochemical properties are consistent with those of the mixture of compounds. The equilibrium position depends on the intramolecular chemical characteristics. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone form is dominant; while in phenols, the enol form is dominant. This invention encompasses all tautomeric forms of the compounds.

[0073] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium. 2 H), tritium ( 3 H), Iodine-125 ( 125 I) or C-14 14 C). All isotopic variations of the compounds of the present invention, regardless of radioactivity, are included within the scope of the present invention.

[0074] For pharmaceuticals or pharmacologically active agents, the term "effective amount" or "therapeutic effective amount" refers to a sufficient quantity of a drug or agent that is non-toxic but achieves the desired effect. For the oral dosage forms of this invention, the "effective amount" of one active substance in the composition refers to the quantity required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. A suitable effective amount in any given case can be determined by a person skilled in the art through routine testing.

[0075] The terms “active ingredient,” “therapeutic agent,” “active substance,” or “active agent” refer to a chemical entity that can effectively treat a target disorder, disease, or symptom.

[0076] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, including deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is a ketone group (i.e., =O), it means that two hydrogen atoms are replaced. Ketone substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents can be arbitrary on a chemically feasible basis.

[0077] The term "C1-C6 alkyl" should be understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers. In particular, the group has 1, 2 or 3 carbon atoms (“C1-C3 alkyl”), such as methyl, ethyl, n-propyl or isopropyl.

[0078] The term "C1-C6 alkyl-O-" should be understood as an alkyl group connected to the rest of the molecule via an oxygen atom, where "C1-C6 alkyl" has the above definition. Examples include methyl-O- and ethyl-O-.

[0079] The term "C3-C6 cycloalkyl" should be understood as referring to a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms, including fused or bridged polycyclic systems. Examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0080] The terms “4-8 membered heterocyclic group” or “4-8 membered heterocyclic alkyl group” should be understood to mean a saturated, unsaturated, or partially saturated monocyclic, bicyclic, or tricyclic ring having 4 to 8 atoms, wherein 1, 2, 3, 4, or 5 ring atoms are selected from N, O, and S, and unless otherwise specified, they may be linked by carbon or nitrogen, wherein -CH 2- The group may optionally be replaced by -C(O)-; and unless otherwise stated to the contrary, the cyclic nitrogen atom or cyclic sulfur atom may optionally be oxidized to form an N-oxide or S-oxide, or the cyclic nitrogen atom may optionally be quaternized; wherein the -NH in the ring may optionally be replaced by an acetyl, formyl, methyl, or methanesulfonyl group; and the ring may optionally be replaced by one or more halogens. It should be understood that when the total number of S and O atoms in the heterocyclic group exceeds 1, these heteroatoms are not adjacent to each other. If the heterocyclic group is bicyclic or tricyclic, at least one ring may optionally be a heteroaromatic ring or an aromatic ring, provided that at least one ring is non-heteroaromatic. If the heterocyclic group is monocyclic, it is necessarily not aromatic. Examples of heterocyclic groups include, but are not limited to, piperidinyl, N-acetylpiperidinyl, N-methylpiperidinyl, N-formylpiperazinyl, N-methanesulfonylpiperazinyl, homopiperazinyl, piperazinyl, azacyclic butyl, oxacyclic butyl, morpholinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, dihydroindolyl, tetrahydropyranyl, dihydro-2H-pyranyl, tetrahydrofuranyl, tetrahydrothiaranyl, tetrahydrothiaran-1-oxide, tetrahydrothiaran-1,1-dioxide, 1H-pyridin-2-one, and 2,5-dioxoimidazolyl.

[0081] The term "4-8 membered heterocyclic alkenyl" should be understood as a non-aromatic monocyclic or polycyclic group containing 4 to 8 ring atoms, preferably 5 to 6 ring atoms, wherein the 4-8 membered heterocyclic alkenyl contains 1 to 3 heteroatoms selected from N, O, S, and P and contains at least one carbon-carbon double bond or carbon-nitrogen double bond. The inclusion of nitro, oxo, or thio in the group name refers to at least one nitrogen, oxygen, or sulfur atom as a ring atom, respectively. The nitrogen or sulfur atom of the 4-8 membered heterocyclic alkenyl may optionally be oxidized to the corresponding N-oxide, S-oxide, or S-dioxide. Preferred 4-8-membered heterocyclic alkenyl groups include, but are not limited to, 1,2,3,4-tetrahydropyridinyl, 1,2-dihydropyridinyl, 1,4-dihydropyridinyl, 1,2,3,6-tetrahydropyridinyl, 1,4,5,6-tetrahydropyrimidinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2-imidazolinyl, 2-pyrazolinyl, dihydroimidazolinyl, dihydrooxazolyl, dihydrooxadiazolyl, dihydrothiazolinyl, 3,4-dihydro-2H-pyranyl, dihydrofuranyl, fluorodihydrofuranyl, and their oxides. "4-8-membered heterocyclic alkenyl groups" may also include those in which two available hydrogen atoms on the same carbon atom of the ring are simultaneously substituted by a single group =O (i.e., forming a carbonyl group).

[0082] The term "5-8 aryl" should be understood as a monocyclic, bicyclic, or tricyclic hydrocarbon ring having 5-8 carbon atoms, exhibiting monovalent aromatic or partially aromatic properties, particularly a ring having 6 carbon atoms ("C6 aryl"), such as a phenyl group; when the 5-8 aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on its substitution site; for example, it can be ortho, para, or meta substituted.

[0083] The term "5-8-membered heteroaryl" should be understood as a monovalent monocyclic, bicyclic, or tricyclic aromatic ring group having 5-8 ring atoms, particularly 5 or 6 carbon atoms, and comprising 1-5 heteroatoms independently selected from N, O, and S. Preferably, it comprises 1-3 monovalent monocyclic, bicyclic, or tricyclic aromatic ring groups independently selected from N, O, and S, and in each case, it may be benzofused. Specifically, the heteroaryl group is selected from thiophene, furanyl, pyrrole, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl, etc.; or pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc.; or borazinyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphridinyl, pteridinyl, carbazolyl, acridineyl, phenazinyl, phenothiazinyl, phenothiazinyl, phenothiazinyl, etc.

[0084] The terms "halogen" or "halogen" refer to fluorine, chlorine, bromine, and iodine.

[0085] Furthermore, it should be noted that, unless otherwise explicitly stated, the descriptive phrase "...independently" used in this invention should be interpreted broadly, meaning that the described individuals are independent of each other and can independently be the same or different specific groups. More specifically, the descriptive phrase "...independently" can mean either that the specific options expressed by the same symbols in different groups do not affect each other, or that the specific options expressed by the same symbols in the same group do not affect each other.

[0086] Beneficial effects

[0087] According to an embodiment of the present invention, the present invention:

[0088] 1) It provides P2X3 antagonists with novel structures, excellent pharmacokinetic properties, and good efficacy or drug-likeness, which can be used to effectively treat P2X3-related diseases and symptoms;

[0089] 2) According to embodiments of the present invention, compared with the control compound, the compounds of the present invention exhibit superior antagonistic activity against human P2X3; the results of rat pharmacokinetic studies show that the compounds of the present invention exhibit excellent pharmacokinetic properties, especially compounds 5 and 7, whose pharmacokinetic properties are significantly improved compared with the positive control; the results of mouse pharmacokinetic studies show that the compounds of the present invention exhibit excellent pharmacokinetic properties, especially compound 13, whose pharmacokinetic properties are significantly improved compared with the positive control;

[0090] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Detailed Implementation

[0091] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0092] Unless otherwise specified, the structures of the compounds in this invention were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts are in units of 10⁻¹⁰. -6 (ppm). The solvents used for NMR determination were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard was tetramethylsilane (TMS).

[0093] The abbreviations of this invention are defined as follows:

[0094] M: Molar concentration, such as 1M hydrochloric acid, which represents a 1 mol / L hydrochloric acid solution.

[0095] LC-MS: Liquid chromatography-mass spectrometry

[0096] DMSO: Dimethyl sulfoxide

[0097] HEPES: 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid

[0098] TLC: Thin-layer chromatography

[0099] Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

[0100] IC 50 The half-maximum inhibitory concentration (MCI) is the concentration at which half of the maximum inhibitory effect is achieved.

[0101] Example 1: Preparation of target compound 1

[0102] (R)-3-(5-methylthiazolyl-2-yl)-5-(morpholin-4-carbonyl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 1)

[0103]

[0104] The synthetic route for target compound 1 is shown below:

[0105]

[0106] Step 1: Synthesis of dimethyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)isophthalate (1B)

[0107]

[0108] At room temperature, Pd(dppf)Cl2 (2.68 g, 3.66 mmol) was added to a solution of 1,4-dioxane (150 mL) containing dimethyl 5-bromoisophthalate (1A) (20 g, 73.2 mmol), pinacol diboronate (20.46 g, 81 mmol), and potassium acetate (21.56 g, 220 mmol). After purging with nitrogen three times, the reaction was carried out at 90 °C for 16 h. TLC showed that the starting material reacted completely. After cooling the system to room temperature, the mixture was filtered, the filtrate was concentrated to dryness, and the residue was purified by silica gel column chromatography to obtain the product 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)dimethyl isophthalate (1B) (13 g, yield 55.4%).

[0109] Step 2: Synthesis of dimethyl 5-(5-methylthiazolyl-2-yl)isophthalate (1C)

[0110]

[0111] At room temperature, Pd(dppf)Cl2 (1.714 g, 2.343 mmol) was added to a mixed solution of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)dimethyl isophthalate (1B) (5 g, 15.62 mmol), 2-bromo-5-methylthiazole (3.34 g, 18.74 mmol), potassium carbonate (5.18 g, 37.5 mmol) in THF (60 mL) and water (12 mL), and reacted at 90 °C for 16 h. The mixture was diluted with water (30 mL), extracted with ethyl acetate (40 mL × 3), concentrated to dryness, and the residue was purified by silica gel column chromatography to give a yellow solid, 5-(5-methylthiazole-2-yl)dimethyl isophthalate (1C) (2.1 g, yield 46.2%).

[0112] LC-MS, M / Z: 292.0 [M+H] + .

[0113] Step 3: Synthesis of 3-(methoxycarbonyl)-5-(5-methylthiazol-2-yl)benzoic acid (1D)

[0114]

[0115] Dimethyl 5-(5-methylthiazol-2-yl)isophthalate (1C) (2 g, 6.87 mmol) was dissolved in a mixed solution of THF (30 mL), water (10 mL), and MeOH (10 mL) at room temperature. 1M sodium hydroxide aqueous solution (6.8 mL, 6.8 mmol) was slowly added dropwise, and the reaction was allowed to proceed overnight at room temperature. Subsequently, the solution was concentrated under reduced pressure to remove methanol and tetrahydrofuran, extracted with ethyl acetate (30 mL), and the aqueous phase was adjusted to pH 4 with 2M hydrochloric acid. It was then extracted with dichloromethane (20 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give a white solid 3-(methoxycarbonyl)-5-(5-methylthiazol-2-yl)benzoic acid (1D) (1.2 g, 63% yield).

[0116] LC-MS, M / Z: 278.3 [M+H] + .

[0117] Step 4: Synthesis of (R)-3-(5-methylthiazolyl-2-yl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl)methyl benzoate (1E)

[0118]

[0119] Under ice bath conditions, 1-propylphosphonic anhydride (6.88 g) was added dropwise to a solution of 3-(methoxycarbonyl)-5-(5-methylthiazolyl-2-yl)benzoic acid (1D) (1.2 g, 4.33 mmol), (R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl-1-amine hydrochloride (1.18 g, 5.19 mmol), N,N-diisopropylethylamine (2.8 g, 21.64 mmol) in N,N-dimethylformamide (20 mL). 10.82 mmol, 50% N,N-dimethylformamide solution), stirred overnight at room temperature, diluted with 50 mL of water, extracted with ethyl acetate (20 mL × 2), concentrated the organic phase to dryness, and purified the residue by silica gel column chromatography to give a white solid (R)-3-(5-methylthiazolyl-2-yl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl)benzoate (1E) (1.1 g, yield 56.4%).

[0120] LC-MS, M / Z: 451.4 [M+H] + .

[0121] Step 5: Synthesis of (R)-3-(5-methylthiazolyl-2-yl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl)benzoic acid (1F)

[0122]

[0123] At room temperature, lithium hydroxide monohydrate (205 mg, 4.88 mmol) was added to a mixed solution of (R)-3-(5-methylthiazolyl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl)benzoate (1E) (1.1 g, 2.44 mmol) in THF (20 mL), MeOH (5 mL), and water (5 mL), and stirred overnight at room temperature. Subsequently, methanol and tetrahydrofuran were removed by vacuum concentration, and the mixture was extracted with ethyl acetate (20 mL). The aqueous phase was adjusted to pH 4 with 2 M hydrochloric acid and then extracted with dichloromethane (20 mL × 2). The organic phase was dried with anhydrous sodium sulfate, filtered, and concentrated to give a white solid (R)-3-(5-methylthiazolyl-2-yl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl)benzoic acid (1F) (0.96 g, yield 90%).

[0124] LC-MS, M / Z: 437.4 [M+H] +

[0125] Step 6: Synthesis of (R)-3-(5-methylthiazolyl-2-yl)-5-(morpholin-4-carbonyl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (1)

[0126]

[0127] Under ice bath conditions, 1-propylphosphonic anhydride (328 mg, 0.344 mmol) was added to a solution of (R)-3-(5-methylthiazolyl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl)benzoic acid (150 mg, 0.344 mmol), morpholine (0.036 mL, 0.412 mmol), and N,N-diisopropylethylamine (0.317 mL, 1.719 mmol) in N,N-dimethylformamide (6 mL). 516 mmol, 50% N,N-dimethylformamide solution), stirred overnight at room temperature, diluted with 10 mL of water, extracted with ethyl acetate (10 mL × 2), concentrated the organic phase to dryness, and purified the residue by silica gel plate separation to give a white solid (R)-3-(5-methylthiazolyl-2-yl)-5-(morpholino-4-carbonyl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (1) (101.3 mg, yield 58.3%).

[0128] 1 H NMR(400MHz,DMSO-d6)δ9.25(d,1H),9.10(s,2H),8.38(s,1H),7.96(d,2H), 7.65(d,1H),5.40-5.20(m,1H),3.75-3.35(m,8H),2.48(s,3H),1.59(d,3H).

[0129] LC-MS, M / Z: 506.3 [M+H] + .

[0130] Example 2: Preparation of target compound 2

[0131] (R)-3-(4-methylpiperazin-1-carbonyl)-5-(5-methylthiazolyl-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 2)

[0132]

[0133] The synthetic route for target compound 2 is shown below:

[0134]

[0135] The synthesis method is described in Example 1.

[0136] 1H NMR(400MHz,DMSO-d6)δ9.29(d,1H),9.13(s,2H),8.41(t,1H),8.00-7.95(m,2H),7.69(d,1H),5.37-5.27(m ,1H),3.66(s,2H),3.42-3.37(m,1H),3.30-3.23(m,1H),2.53(d,3H),2.38(d,4H),2.23(s,3H),1.62(d,3H).

[0137] LC-MS, M / Z: 519.3 [M+H] + .

[0138] Example 3: Preparation of target compound 3

[0139] (R)-3-(3-hydroxyazacyclobutane-1-carbonyl)-5-(5-methylthiazolyl-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 3)

[0140]

[0141] The synthetic route for target compound 3 is shown below:

[0142]

[0143] The synthesis method is described in Example 1.

[0144] 1 H NMR(400MHz,DMSO-d6)δ9.33(d,1H),9.14(s,2H),8.46(t,1H),8.19-8.15(m,2H),7.69(d,1H),5.81(d,1H),5.38- 5.28(m,1H),4.58-4.44(m,2H),4.37-4.25(m,1H),4.12-4.05(m,1H),3.88-3.80(d,1H),2.53(d,3H),1.63(d,3H).

[0145] LC-MS, M / Z: 492.2 [M+H] + .

[0146] Example 4: Preparation of target compound 4

[0147] 3-((3S,5S)-3,5-dimethylmorpholino-4-carbonyl)-5-(5-methylthiazolyl-2-yl)-N-((R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 4)

[0148]

[0149] The synthetic route for target compound 4 is shown below:

[0150]

[0151] The synthesis method is described in Example 1.

[0152] 1 H NMR(400MHz, CDCl3)δ8.95(s,2H),8.28(t,1H),8.08(t,1H),7.89(t,1H),7.55(d,1H),6.95(d, 1H),5.42-5.32(m,1H),3.95-3.81(m,4H),3.50(dd,2H),2.55(d,3H),1.74(d,3H),1.30(d,6H).

[0153] LC-MS, M / Z: 534.3 [M+H] + .

[0154] Example 5: Preparation of target compound 5

[0155] (R)-3-(3-hydroxy-3-methylazacyclobutane-1-carbonyl)-5-(5-methylthiazolyl-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 5)

[0156]

[0157] The synthetic route for target compound 5 is shown below:

[0158]

[0159] The synthesis method is described in Example 1.

[0160] 1 H NMR(400MHz, CDCl3)δ8.97(d,2H),8.30(d,1H),8.11(d,1H),7.98(d,1H),7.79(dd,1H),7.49(s ,1H),5.42-5.31(m,1H),4.35-4.05(m,4H),3.55(s,1H),2.51(s,3H),1.72(s,3H),1.54(s,3H).

[0161] LC-MS, M / Z: 506.2 [M+H] + .

[0162] Example 6: Preparation of target compound 6

[0163] 3-((S)-3-methylmorpholino-4-carbonyl)-5-(5-methylthiazolyl-2-yl)-N-((R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 6)

[0164]

[0165] The synthetic route for target compound 6 is shown below:

[0166]

[0167] The synthesis method is described in Example 1.

[0168] 1 H NMR(400MHz, CDCl3)δ8.97(s,2H),8.22(s,1H),7.93(s,1H),7.73(s,1H),7.53(d,1H),7.37 -7.29(m,1H),5.41-5.32(m,1H),4.20-3.20(m,7H),2.54(d,3H),1.73(d,3H),1.41(d,3H).

[0169] LC-MS, M / Z: 520.2 [M+H] + .

[0170] Example 7: Preparation of target compound 7

[0171] 3-((R)-3-methylmorpholino-4-carbonyl)-5-(5-methylthiazolyl-2-yl)-N-((R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (Target compound 7)

[0172]

[0173] The synthetic route for target compound 7 is shown below:

[0174]

[0175] The synthesis method is described in Example 1.

[0176] 1H NMR(400MHz, CDCl3)δ8.97(s,2H),8.22(s,1H),7.92(s,1H),7.73(s,1H),7.53(d,1H),7 .37(d,1H),5.41-5.32(m,1H),4.20-3.20(m,7H),2.54(d,3H),1.73(d,3H),1.40(d,3H).

[0177] LC-MS, M / Z: 520.2 [M+H] + .

[0178] Example 8: Preparation of target compound 8

[0179] (R)-3-(5-methylthiazolyl-2-yl)-5-(2-oxa-6-azaspiro[3.3]heptane-6-carbonyl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 8)

[0180]

[0181] The synthetic route for target compound 8 is shown below:

[0182]

[0183] The synthesis method is described in Example 1.

[0184] 1 H NMR(400MHz, CDCl3)δ8.96(s,2H),8.35(t,1H),8.20(t,1H),8.01(t,1H),7.53(d,1H),7.18 (d,1H),5.41-5.33(m,1H),4.81(d,4H),4.51(s,2H),4.37(s,2H),2.54(d,3H),1.73(d,3H).

[0185] LC-MS, M / Z: 518.2 [M+H] + .

[0186] Example 9: Preparation of target compound 9

[0187] 3-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane-5-carbonyl)-5-(5-methylthiazolyl-2-yl)-N-((R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 9)

[0188]

[0189] The synthetic route for target compound 9 is shown below:

[0190]

[0191] The synthesis method is described in Example 1.

[0192] 1 H NMR(400MHz,DMSO-d6)δ9.30(d,1H),9.14(s,2H),8.44(d,1H),8.14-8.05(m,2H),7.69(d,1H),5.36-5.31(m,1H),4.40-4.89(m,2H),3.9 5-3.87(m,1H),3.80-3.69(m,1H),3.56-3.54(m,1H),3.33-3.31(m,1H),2.52(q,3H),2.01-1.91(m,1H),1.92-1.77(m,1H),1.64(t,3H).

[0193] LC-MS, M / Z (ESI): 518.53 [M+H] + .

[0194] Example 10: Preparation of target compound 10

[0195] 3-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-carbonyl)-5-(5-methylthiazo-2-yl)-N-((R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 10)

[0196]

[0197] The synthetic route for target compound 10 is shown below:

[0198]

[0199] The synthesis method is described in Example 1.

[0200] 1H NMR(400MHz,DMSO-d6)δ9.30(d,1H),9.14(s,2H),8.44(d,1H),8.14-8.05(m,2H),7.69(d,1H),5.36-5.31(m,1H),4.40-4.89(m,2H),3.9 5-3.87(m,1H),3.80-3.69(m,1H),3.56-3.54(m,1H),3.33-3.31(m,1H),2.52(q,3H),2.01-1.91(m,1H),1.92-1.77(m,1H),1.64(t,3H).

[0201] LC-MS, M / Z (ESI): 518.53 [M+H] + .

[0202] Example 11: Preparation of target compound 11

[0203] (R)-3-(5-methylthiazolyl-2-yl)-5-(pyrrolidine-1-ylsulfonyl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (Target compound 11)

[0204]

[0205] The synthetic route for target compound 11 is shown below:

[0206]

[0207] Step 1: Synthesis of methyl 3-bromo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate (11B)

[0208]

[0209] At room temperature, Pd(dppf)Cl2 (0.644 g, 0.88 mmol) was added to a solution of methyl 3-bromo-5-iodobenzoate (6 g, 17.6 mmol), pinacol diboronate (5.81 g, 22.88 mmol), and potassium acetate (5.18 g, 52.8 mmol) in 1,4-dioxane (50 mL). After purging with nitrogen three times, the mixture was reacted at 80 °C for 16 h. The system was cooled to room temperature and filtered. The filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography to obtain the product methyl 3-bromo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate (3 g, 50% yield).

[0210] Step 2: Synthesis of methyl 3-bromo-5-(5-methylthiazolyl-2-yl)benzoate (11C)

[0211]

[0212] At room temperature, Pd(dppf)Cl2 (0.644 g, 0.88 mmol) was added to a mixed solution of methyl 3-bromo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate (3 g, 8.8 mmol), 2-bromo-5-methylthiazole (2.35 g, 13.2 mmol), potassium carbonate (3.04 g, 21.99 mmol) in THF (30 mL) and water (8 mL), and the reaction was carried out at 90 °C for 16 h. The mixture was diluted with water (20 mL), extracted with ethyl acetate (30 mL × 3), concentrated to dryness, and the residue was purified by silica gel column chromatography to give the product methyl 3-bromo-5-(5-methylthiazole-2-yl)benzoate (1.8 g, yield 65.5%).

[0213] Step 3: Synthesis of methyl 3-((4-methoxybenzyl)thio)-5-(5-methylthiazolyl-2-yl)benzoate (11D)

[0214]

[0215] Methyl 3-bromo-5-(5-methylthiazol-2-yl)benzoate (1.8 g, 5.77 mmol), (4-methoxyphenyl)methanethiol (1.156 g, 7.50 mmol), tris(dibenzylacetone)dipalladium (1.056 g, 1.153 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (0.667 g, 1.153 mmol), and N,N-diisopropylethylamine (1.49 g, 11.53 mmol) were added. The methyl benzoate (1.6 g, yield 72%) was dissolved in 20 mL of dioxane, purged with nitrogen three times, reacted at 110 °C for 12 hours, diluted with water (20 mL), extracted with ethyl acetate (20 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the concentrate was purified by column chromatography (petroleum ether: ethyl acetate (V:V) = 3:1) to give the title compound methyl 3-((4-methoxybenzyl)thio)-5-(5-methylthiazolyl-2-yl)benzoate (1.6 g, yield 72%).

[0216] LC-MS, M / Z: 386.1 [M+H] +

[0217] Step 4: Synthesis of methyl 3-(chlorosulfonyl)-5-(5-methylthiazolyl-2-yl)benzoate (11E)

[0218]

[0219] Methyl 3-((4-methoxybenzyl)thio)-5-(5-methylthiazol-2-yl)benzoate (1 g, 2.59 mmol) was dissolved in 5 mL of water and 5 mL of acetic acid. Dichlorohydantoin (756 mg, 3.89 mmol) was slowly added, and the mixture was stirred at room temperature for 12 hours. The solution was diluted with water (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give the title compound methyl 3-(chlorosulfonyl)-5-(5-methylthiazol-2-yl)benzoate (600 mg, 69% yield).

[0220] Step 5: Methyl 3-(5-methylthiazolyl-2-yl)-5-(pyrrolidine-1-ylsulfonyl)benzoate (11F)

[0221]

[0222] Methyl 3-(chlorosulfonyl)-5-(5-methylthiazol-2-yl)benzoate (300 mg, 0.904 mmol) was dissolved in 4 mL of dichloromethane, and triethylamine (274 mg, 2.71 mmol) and tetrahydropyrrole (96 mg, 1.356 mmol) were added. The mixture was stirred at room temperature for 1 hour. After the reaction was completed, 10 mL of water was added for dilution, and the mixture was extracted with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound methyl 3-(5-methylthiazol-2-yl)-5-(pyrrolidine-1-ylsulfonyl)benzoate (200 mg, 60% yield).

[0223] LC-MS, M / Z: 367.1 [M+H] +

[0224] Step 6: 3-(5-methylthiazolyl-2-yl)-5-(pyrrolidine-1-ylsulfonyl)benzoic acid (11G)

[0225]

[0226] Methyl 3-(5-methylthiazol-2-yl)-5-(pyrrolidine-1-ylsulfonyl)benzoate (140 mg, 0.382 mmol) was dissolved in 5 mL of methanol, and sodium hydroxide (76 mg, 1.91 mmol) was added. The mixture was stirred at room temperature for 3 hours. The solution was evaporated to dryness, and water (20 mL) was added. The pH was adjusted to 1 with 1 M hydrochloric acid, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined and evaporated to dryness to give compound 3-(5-methylthiazol-2-yl)-5-(pyrrolidine-1-ylsulfonyl)benzoic acid (100 mg, 74%).

[0227] LC-MS, M / Z: 353.1 [M+H] +

[0228] Step 7: Synthesis of (R)-3-(5-methylthiazolyl-2-yl)-5-(pyrrolidine-1-ylsulfonyl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (11)

[0229]

[0230] 3-(5-methylthiazolyl-2-yl)-5-(pyrrolidine-1-ylsulfonyl)benzoic acid (100 mg, 0.284 mmol), (R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethylamine (65.1 mg, 0.34 mmol), 1-propylphosphonic anhydride (542 mg, 0.851 mmol, 50% N,N-dimethylformamide solution), and N,N-diisopropylethylamine (110 mg, 0.851 mmol) were dissolved in 2 mL of N,N-dimethylformamide and stirred at room temperature for 12 hours. Dilute with water (20 mL), extract with ethyl acetate (20 mL × 3), combine the organic phases, dry to anhydrous sodium sulfate, concentrate, and purify the concentrate by column chromatography (petroleum ether: ethyl acetate (V:V) = 3:1) to give the title compound (R)-3-(5-methylthiazolyl-2-yl)-5-(pyrrolidine-1-ylsulfonyl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (55 mg, yield 37%).

[0231] 1 H NMR(400MHz, CDCl3)δ9.50(d,1H),9.12(s,2H),8.55(s,1H),8.40(s,2H),7.75 (s,1H),5.32(t,1H),3.25(s,3H),3.20(t,2H),2.52(s,3H),1.60-1.70(m,6H)

[0232] LC-MS, M / Z: 526.1 [M+H] + .

[0233] Example 12: Preparation of target compound 12

[0234] (R)-3-(5-methylthiazolyl-2-yl)-5-(2-oxopyrrolidone-1-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 12)

[0235]

[0236] The synthetic route for target compound 12 is shown below:

[0237]

[0238] Step 1: Synthesis of methyl 3-bromo-5-(2-oxopyrrolidone-1-yl)benzoate (12B)

[0239]

[0240] Under nitrogen protection, a solution of methyl 3-bromo-5-iodobenzoate (10 g, 29.3 mmol), 2-pyrrolidone (3.74 g, 44.0 mmol), cesium carbonate (14.33 g, 44.0 mmol), and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (2.55 g, 4.40 mmol) in 1,4-dioxane (100 mL) was added to tris(dibenzylacetone)dipalladium (1.343 g, 1.467 mmol), and stirred overnight at 55 °C. The solution was diluted with water (200 mL), extracted with ethyl acetate (100 mL × 2), concentrated to dryness, and the residue was purified by silica gel column chromatography to obtain solid methyl 3-bromo-5-(2-oxopyrrolidone-1-yl)benzoate (6 g, yield 68.6%).

[0241] LC-MS, M / Z: 298.0 [M+H] +

[0242] Step 2: Synthesis of methyl 3-(2-oxopyrrolidone-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate (12C)

[0243]

[0244] At room temperature, Pd(dppf)Cl2 (0.37 g, 0.503 mmol) was added to a solution of methyl 3-bromo-5-(2-oxopyrrolidone-1-yl)benzoate (3 g, 10.06 mmol), pinacol diboronate (2.81 g, 11.07 mmol), and potassium acetate (2.47 g, 25.2 mmol) in 1,4-dioxane (30 mL). After purging with nitrogen three times, the mixture was reacted at 90 °C for 16 h. The system was cooled to room temperature and filtered. The filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography to obtain the product methyl 3-(2-oxopyrrolidone-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzoate (0.7 g, yield 20.2%).

[0245] Step 3: Synthesis of methyl 3-(5-methylthiazolyl-2-yl)-5-(2-oxopyrrolidine-1-yl)benzoate (12D)

[0246]

[0247] At room temperature, methyl 3-(2-oxopyrrolidone-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron-2-yl)benzoate (0.7 g, 2.03 mmol), 2-bromo-5-methylthiazole (0.44 g, 2.44 mmol), potassium carbonate (0.67 g, 4.87 mmol) in THF (10 mL) and water (3 mL) was added to Pd(dppf)Cl2 (0.148 g, 0.203 mmol), and the reaction was carried out at 90 °C for 16 h. The mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL × 3), concentrated to dryness, and the residue was purified by silica gel column chromatography to give the product methyl 3-(5-methylthiazole-2-yl)-5-(2-oxopyrrolidone-1-yl)benzoate (0.27 g, yield 42.1%).

[0248] LC-MS, M / Z: 317.2 [M+H] + .

[0249] Step 4: Synthesis of 3-(5-methylthiazolyl-2-yl)-5-(2-oxopyrrolidone-1-yl)benzoic acid (12E)

[0250]

[0251] Methyl 3-(5-methylthiazol-2-yl)-5-(2-oxopyrrolidone-1-yl)benzoate (0.27 g, 0.853 mmol) was dissolved in a mixture of THF (8 mL), water (2 mL), and MeOH (2 mL) at room temperature. Lithium hydroxide monohydrate (71.6 mg, 1.707 mmol) was added, and the reaction was carried out overnight at room temperature. Subsequently, methanol and tetrahydrofuran were removed by concentration under reduced pressure. The pH was adjusted to approximately 4 with 2 M hydrochloric acid, and the mixture was extracted with dichloromethane (10 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give a white solid 3-(5-methylthiazol-2-yl)-5-(2-oxopyrrolidone-1-yl)benzoic acid (0.19 g, yield 73.6%).

[0252] LC-MS, M / Z: 303.2 [M+H] +

[0253] Step 5: Synthesis of (R)-3-(5-methylthiazolyl-2-yl)-5-(2-oxopyrrolidone-1-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (target compound 12)

[0254]

[0255] Under ice bath conditions, 1-propylphosphonic anhydride (1.0 g, 0.628 mmol) was added dropwise to a solution of 3-(5-methylthiazolyl-2-yl)-5-(2-oxopyrrolidone-1-yl)benzoic acid (0.19 g, 0.628 mmol), (R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl-1-amine hydrochloride (131 mg, 0.754 mmol), N,N-diisopropylethylamine (406 mg, 3.14 mmol) in N,N-dimethylformamide (10 mL). 1.57 mmol (50% N,N-dimethylformamide solution), stirred overnight at room temperature, diluted with 30 mL of water, extracted with ethyl acetate (20 mL × 2), the organic phase was condensed to dryness, and the residue was purified by silica gel plate separation. White solid (R)-3-(5-methylthiazolyl-2-yl)-5-(2-oxopyrrolidone-1-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (12) (47.4 mg, yield 15.9%).

[0256] 1 H NMR(400MHz, CDCl3)δ9.00(s,2H),8.27(t,1H),7.90(t,1H),7.84(t,1H),7.48(d,1H),7.35(d,1H ),5.42-5.32(m,1H),3.91-3.77(m,2H),2.67(t,2H),2.52(d,3H),2.25-2.12(m,2H),1.76(d,3H).

[0257] LC-MS, M / Z: 476.2 [M+H] +

[0258] Example 13: Preparation of target compound 13

[0259] (R)-3-(2-hydroxypropyl-2-yl)-5-(5-methylthiazolyl-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (Target compound 13)

[0260]

[0261] The synthetic route for target compound 13 is shown below:

[0262]

[0263] Under nitrogen protection, a THF (5 mL) solution of (R)-3-(5-methylthiazolyl-2-yl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl)benzoate (60 mg, 0.133 mmol) was cooled to 0 °C, and methyl magnesium bromide (0.266 mL, 0.799 mmol, 3 M THF solution) was slowly added dropwise. The system was then brought to room temperature and reacted overnight. The reaction was quenched with saturated ammonium chloride solution (10 mL), extracted with ethyl acetate (10 mL × 2), concentrated to dryness, and the residue was purified by silica gel plate separation to give a white solid (R)-3-(2-hydroxypropyl-2-yl)-5-(5-methylthiazolyl-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (13) (13.6 mg, yield 22.7%).

[0264] 1 H NMR(400MHz, CDCl3)δ8.94(s,2H),8.14(d,2H),7.97(s,1H),7.52(s,1H),6.83 (d,1H),5.43-5.33(m,1H),2.54(s,3H),2.01(s,1H),1.72(d,3H),1.63(s,6H).

[0265] LC-MS, M / Z: 451.2 [M+H] + .

[0266] Example 14: Control compound 1 and its preparation

[0267]

[0268] The control compound 1 was synthesized with reference to patent application WO 2016 / 091776.

[0269] The “control compound 1” mentioned below refers to the compound described in Example 14.

[0270] Example 15: Control compound 2 and its preparation

[0271]

[0272] The control compound 2 was synthesized with reference to patent application WO 2016 / 091776.

[0273] The term "comparison compound 2" as used below refers to the compound described in Example 15.

[0274] Test Example 1: FLIPR method for determining the antagonistic activity of hP2X3 antagonists against hP2X3

[0275] The antagonistic activity of human P2X3 receptor (hP2X3) antagonists against hP2X3 was evaluated using the FLIPR Calcium4Assay Kit (Molecular Devices, R8141) and the FLIPR TETRA instrument (MolecularDevices, 0296) to detect calcium flow signals. Twenty-four hours before the experiment, human cells stably transfected with the hP2X3 receptor were cultured at a rate of 2 × 10⁻⁶ cells / year. 5 Cells were seeded at a density of 50 μL / mL in 384-well plates and incubated at 37°C for 16–24 h in a 5% CO2 incubator. 500 nmol of the desired concentration of the test compound was prepared using DMSO stock solution (20–50 mM DMSO), and added to each well of the 384-well plate. 30 μL of FLIPR Assay buffer (containing 1.26 mM Ca) was then added to the top of each well. 2+ Mix 1×HBSS + 2mM CaCl2 + 20mM HEPES, shake for 20-40 min to mix. Prepare 3 times the required concentration of agonist (α,β-meATP) using FLIPR Assay buffer (final concentration required 400 nM), and add 45 μL of agonist to each well of another 384-well plate. Take the cell culture plate from the previous day, aspirate the cell supernatant, and add 30 μL of Dye to each well. The cells were incubated for 1 hour using a Calcium 4 Assay Kit (diluted with FLIPR buffer). 15 μL of the compound was added to each well (using a FLIPR instrument). After 15 minutes, 22.5 μL of the agonist was added to each well, and the fluorescence signal was detected (excitation wavelength 470 nm-495 nm, emission wavelength 515 nm-575 nm). The difference between the peak and trough values ​​was used as the baseline data. The highest concentration of the positive control was taken as the 100% inhibition rate, and DMSO data as the 0% inhibition rate. The inhibition effect curve of the compound was fitted using GraphpadPrism 6 software, and the IC50 was calculated. 50 value.

[0276] Table 1 shows the antagonistic activity of the tested compounds against hP2X3.

[0277]

[0278] Experimental results show that, compared with the control compound, the compound of the present invention exhibits superior antagonistic activity against human P2X3.

[0279] Test Example 2: Rat Pharmacokinetic Test

[0280] Pharmacokinetic studies were conducted on rats using three male SD rats (180-240g), which were fasted overnight and administered 10mg / kg orally via gavage. Blood samples were collected before administration and at 15, 30 minutes, 1, 2, 4, 8, and 24 hours after administration. Blood samples were centrifuged at 8000 rpm for 6 minutes at 4°C, and plasma was collected and stored at -20°C. Plasma samples at each time point were mixed with 3-5 times the volume of acetonitrile solution containing an internal standard, vortexed for 1 minute, centrifuged at 13000 rpm for 10 minutes at 4°C, and the supernatant was mixed with 3 times the volume of water. An appropriate amount of the mixture was analyzed by LC-MS / MS. Key pharmacokinetic parameters were analyzed using a non-compartmental model with WinNonlin 7.0 software.

[0281] Table 2. Rat pharmacokinetics of the tested compounds

[0282]

[0283] The results of the pharmacokinetic studies in rats showed that the compounds of the present invention exhibited excellent pharmacokinetic properties, especially compounds 5 and 7, which showed significantly improved pharmacokinetic properties compared with the positive control.

[0284] Test Example 3: Mouse Pharmacokinetic Study

[0285] Pharmacokinetic studies were conducted in mice using male ICR mice (20-25g, fasted overnight). Three mice were administered 10mg / kg orally via gavage. Blood samples were collected before administration and at 15, 30 minutes, 1, 2, 4, 8, and 24 hours after administration. 6800g of blood samples were centrifuged at 2-8℃ for 6 minutes, and plasma was collected and stored at -80℃. Plasma samples from each time point were mixed with 3-5 times the volume of acetonitrile solution containing an internal standard, vortexed for 1 minute, centrifuged at 13000 rpm at 4℃ for 10 minutes, and the supernatant was mixed with 3 times the volume of water. An appropriate amount of the mixture was analyzed by LC-MS / MS. Key pharmacokinetic parameters were analyzed using a non-compartmental model with WinNonlin 7.0 software.

[0286] Table 3. Pharmacokinetics of the tested compounds in mice.

[0287]

[0288] The results of the pharmacokinetic studies in mice showed that the compounds of the present invention exhibited excellent pharmacokinetic properties, especially compound 13, which showed significantly improved pharmacokinetic properties compared with the positive control.

Claims

1. The compound represented by Formula I and its pharmaceutically acceptable salt: in, L is , Or it may not exist; When L is hour; for The ring A is unsubstituted or replaced by R. a Substituted nitrogen-containing 4-8 membered heterocyclic alkyl groups, the unsubstituted or R a In the substituted nitrogen-containing 4-8 membered heterocyclic alkyl group, the heteroatom is selected from one or more of N, S, O, and P, and the number of heteroatoms is 1-3; the R- a In the substituted 4-8 membered heterocyclic alkyl groups, R a Each substituent is independently selected from one or more of the following substituents: halogen, hydroxyl, cyano, amino, oxo, C1-C6 alkyl, C1-C6 alkyl-O-, C1-C6 alkyl substituted with 1 to 5 identical or different halogens; when there are multiple substituents, the substituents are identical or different; When L is hour; for The ring A is unsubstituted or replaced by R. a Substituted nitrogen-containing 4-8 membered heterocyclic alkyl groups, the unsubstituted or R a In the substituted nitrogen-containing 4-8 membered heterocyclic alkyl groups, the heteroatom is selected from one or more of N, S, and O, and the number of heteroatoms is 1-3; R a Each substituent is independently selected from one or more of the following substituents: halogen, hydroxyl, cyano, amino, oxo, C1-C6 alkyl, C1-C6 alkyl-O-, C1-C6 alkyl substituted with 1 to 5 identical or different halogens; when there are multiple substituents, the substituents are identical or different; The 4-8 membered heterocyclic alkyl group is a 4-6 membered heterocyclic alkyl group; When L does not exist; R 1 Selected from R a Substituted C1-C6 alkyl groups, or for The ring A is R a Substituted nitrogen-containing 4-6 membered heterocyclic alkyl groups; the R- a In the substituted C1-C6 alkyl groups, R a Selected from hydroxyl groups; the ring A is R a In the substituted nitrogen-containing 4-6 membered heterocyclic alkyl groups, R a Selected from oxo compounds, with one heteroatom; And the compound shown in Formula I is not: 、 、 、 。 2. The compound represented by Formula I according to claim 1, and its pharmaceutically acceptable salt, characterized in that: When L is And R 1 For not replaced or by R a When the substituted 4-8-membered heterocyclic alkyl group is used, the 4-8-membered heterocyclic alkyl group is independently aziridine, oxaziridine, tetrahydropyrrolidinyl, morpholinyl, piperazine, tetrahydrofuranyl, 2-oxa-6-azaspiro[3.3]hept-6-yl, 2-oxa-5-azabicyclo[2.2.1]hept-5-yl, hexahydropyran or tetrahydro-2H-thiopyran 1,1-dioxide; And / or, when R 1 For R a When the substituted 4-8 membered heterocyclic alkyl group is used, the number of substitutions is 1-3; And / or, when R 1 For R a When substituted with 4-8 membered heterocyclic alkyl groups, R a When each is an independent halogen, the halogen is F, Cl, Br or I; And / or, when R 1 For R a When substituted with 4-8 membered heterocyclic alkyl groups, R a When each is independently a C1-C6 alkyl group, the C1-C6 alkyl group in the substitution is independently a C1-C4 alkyl group; And / or, when R 1 For R a When substituted with 4-8 membered heterocyclic alkyl groups, R a Each independently undergoes oxygenation; And / or, when R 1 For R a When substituted with 4-8 membered heterocyclic alkyl groups, R a Each is an independent hydroxyl group.

3. The compound represented by Formula I according to claim 1, and its pharmaceutically acceptable salt, characterized in that: When R 1 When the substituted nitrogen-containing 4-8 membered heterocyclic alkyl group is replaced, the number of substituted groups is one or two; And / or, when R 1 For R a When substituted with 4-8 membered heterocyclic alkyl groups, R a When each is an independent halogen, the halogen is F; And / or, when R 1 For R a When substituted with 4-8 membered heterocyclic alkyl groups, R a When each is an independent C1-C6 alkyl group, the C1-C6 alkyl group in the substitution is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl.

4. The compound represented by Formula I according to claim 1, and its pharmaceutically acceptable salt, characterized in that, L is At that time, R 1 for , , , , , , , , or ; When L does not exist, R 1 for C(CH3)2OH or ; L is At that time, R 1 for , , , , or .

5. The compound represented by Formula I according to claim 4, and its pharmaceutically acceptable salt, characterized in that, L is At that time, R 1 for , , , , , , , , , , or ; L is At that time, R 1 for , , , , , or .

6. The compound represented by Formula I according to claim 1, and its pharmaceutically acceptable salt, characterized in that, The compound is the compound in Scheme 1; wherein the compound in Scheme 1 has the structure shown in Formula II: In scheme 1, R 1 Selected from unsubstituted or R a Substituted 4-8 membered heterocyclic alkyl groups, the unsubstituted or R a In the substituted 4-8 membered heterocyclic alkyl groups, R a Each of the following substituents is independently selected from one or more of the following: hydroxyl, C1-C6 alkyl, or C1-C6 alkyl-O-.

7. The compound represented by Formula I according to claim 1, or a pharmaceutically acceptable salt, characterized in that, The compound is the compound in Scheme 2; wherein the compound in Scheme 2 has the structure shown in Formula III; In scheme 2, R 1 It is selected from 4-6 membered heterocyclic alkyl groups, and the heteroatom is selected from one or more of N, S and O, with a heteroatom number of 1-3.

8. The compound represented by Formula I according to claim 1, or a pharmaceutically acceptable salt, characterized in that, The compound is the compound in Scheme 3, wherein the compound in Scheme 3 has the structure shown in Formula IV: ; In scheme 3, R 1 Selected from unsubstituted or R a Substituted C2-C4 alkyl groups; the R-substituted C2-C4 alkyl groups a In the substituted C2-C4 alkyl groups, R a Selected from hydroxyl groups.

9. The compound represented by Formula I according to claim 6, and its pharmaceutically acceptable salt, characterized in that, In Scheme 1, the 4-8 membered heterocyclic alkyl group is independently aziridine, aziridine propane, oxacyclobutane, tetrahydropyrrolidinyl, morpholinyl, piperazine, tetrahydrofuranyl, 2-oxa-6-azaspiro[3.3]hept-6-yl, 2-oxa-5-azabicyclo[2.2.1]hept-5-yl, hexahydropyran, or tetrahydro-2H-thiopyran 1,1-dioxide.

10. The compound represented by Formula I according to claim 7, and its pharmaceutically acceptable salt, characterized in that, In Scheme 2, the 4-6 membered heterocyclic alkyl group is azacyclopropane, azacyclobutane, or tetrahydropyrrolealkyl.

11. The compound of formula I according to claim 1, and its pharmaceutically acceptable salt, characterized in that, for , The ring A is unsubstituted or replaced by R. a Substituted nitrogen-containing 3- or 4-membered heterocyclic alkyl groups, wherein the heteroatom is selected from one or more of N, S, and O, and R a Selected from one or more hydroxyl groups or one or more C1-C3 alkyl groups; Alternatively, ring A may be unsubstituted or replaced by R. a Substituted nitrogen- and oxygen-containing 5- or 6-membered heterocyclic alkyl groups, wherein the heteroatoms are selected from one or more of N, S, and O, and the number is 1 to 3, R a Selected from C1-C3 alkyl groups.

12. The compound of formula I according to claim 11, and its pharmaceutically acceptable salt, characterized in that, The ring A is selected from Among them, at least one of X1, X2, X3, X4, and X5 is selected from O or S, the rest are selected from C or N, and X6 is selected from C1-C3 alkyl; Alternatively, ring A may be a 6-membered spirobi-heterocycle containing nitrogen and oxygen or a 6-membered bridged bi-heterocycle containing nitrogen and oxygen, with 1 to 3 heteroatoms.

13. The compound of formula I according to claim 11, and its pharmaceutically acceptable salt, characterized in that, The ring A is selected from X6 is selected from C1-C3 alkyl groups.

14. The compound of formula I according to claim 1, and its pharmaceutically acceptable salt, characterized in that, The compound represented by Formula I is selected from any of the following compounds: 。 15. A pharmaceutical composition, characterized in that, It comprises a compound of Formula I as described in any one of claims 1-14, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable pharmaceutical carrier or excipient.

16. The pharmaceutical composition according to claim 15, characterized in that, The excipient is a diluent.

17. Use of the compound of Formula I according to any one of claims 1-14, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 15 or 16 in the preparation of a medicament for treating diseases related to P2X3.

18. The use according to claim 17, characterized in that, The P2X3-related diseases are pain, respiratory diseases, or genitourinary diseases.

19. The use according to claim 17, characterized in that, The pain can be classified as chronic pain or acute pain.

20. The use according to claim 17, characterized in that, The pain described is surgical pain.

21. The use according to claim 17, characterized in that, The pain can be caused by endometriosis, neuropathic pain, back pain, cancer pain, inflammatory pain, migraine, or visceral pain.

22. The use according to claim 17, characterized in that, The pain described is pain from endometriosis and neuropathic pain.

23. The use according to claim 17, characterized in that, The urogenital system diseases mentioned include decreased bladder capacity, urge incontinence, stress incontinence, hyperresponsiveness of the bladder, benign prostatic hyperplasia, prostatitis, detrusor hyperreflexia, urinary frequency, nocturia, urinary urgency, overactive bladder, pelvic hypersensitivity, urethritis, pelvic pain syndrome, prostatodynia, cystitis, or idiopathic bladder hypersensitivity.

24. The use according to claim 17, characterized in that, The urogenital system disease mentioned is overactive bladder.

25. The use according to claim 17, characterized in that, The respiratory diseases mentioned include chronic obstructive pulmonary disease, pulmonary hypertension, pulmonary fibrosis, asthma, obstructive sleep apnea, chronic cough, and acute cough.

26. The use according to claim 17, characterized in that, The respiratory disease is refractory chronic cough.

Images