WO2021238834A1

WO2021238834A1 - Arylformamide compound and preparation method and medical use thereof

Info

Publication number: WO2021238834A1
Application number: PCT/CN2021/095424
Authority: WO
Inventors: 殷惠军; 闫旭; 宗利斌; 刘国标; 张守良; 陈彬
Original assignee: 中国医药研究开发中心有限公司
Priority date: 2020-05-25
Filing date: 2021-05-24
Publication date: 2021-12-02
Also published as: TW202144342A; AU2021282039A1; CN114072399A

Abstract

The present invention relates to an arylformamide compound and a preparation method and a medical use thereof. Particularly, the present invention relates to a compound shown in a general formula (I), a preparation method thereof, a pharmaceutical composition containing the compound and a use thereof as a P2X3 receptor antagonist. The compound and the pharmaceutical composition containing the compound can be used for treating and/or preventing diseases related to P2X3 activity, such as chronic cough, pain, endometriosis, overactive bladder etc. The definition of each substituent in the general formula (I) is the same as that in the specification.

Description

Aromatic carboxamide compounds, preparation method and medical use thereof

Technical field

The invention belongs to the technical field of medicine, and specifically relates to an arylformamide compound, a preparation method thereof, and a pharmaceutical composition containing the same, and its use as a P2X3 receptor antagonist in the treatment and/or prevention of diseases related to P2X3 activity use.

Background technique

The elevated level of ATP in the pathological environment indicates that it plays an important role in the pathogenesis of many diseases. In particular, ATP can drive and regulate various sensory behaviors and related responses. When the body is stimulated (ultraviolet rays and chemical damage, etc.) or in a pathological state (asthma, bladder pain syndrome, etc.), ATP has a greater impact on sensation.

Many cell surface receptors (purinergic receptors) are involved in mediating the sensory signal function of ATP. Among them, P2X3 is the main receptor that mediates the sensory effects of ATP. The P2X3 receptor is an ATP-gated cation channel and belongs to the P2X receptor family. The P2X receptor family also includes P2X1, P2X2, P2X4, P2X5, P2X6, and P2X7. P2X3 acts in the body in the form of homotrimer P2X3 or heterotrimer P2X2/3 (NeuroReport, 10, 1107-1111).

P2X3 and P2X2/3 are mainly expressed in the small and medium diameter C- and Aδ-fiber sensory neurons in the dorsal root ganglia (DRG) and cranial sensory ganglia, as well as peripheral nerve endings in the receptive fields of skin, joints and internal organs.

The P2X3 receptor is a member of the purinergic receptor family. It is a non-selective ligand-gated ion channel. After ATP activates it, it allows Na ⁺ , K ⁺ , and Ca ^{2+ to} pass, especially Ca ²⁺ . The permeability is the most obvious, and it plays an important role in the generation and transmission of harmful information. When the body is injured or nerve damage, a large amount of ATP is released, which activates the P2X3 receptor of the presynaptic membrane, causing a large amount of Ca ²⁺ influx, and the increase in intracellular calcium concentration activates protein kinase A (PKA) and protein kinase C ( Protein kinase C (PKC) phosphorylates PKA and PKC, and at the same time promotes the release of glutamate, which further activates the NMDA receptors, leading to the generation of excitatory postsynaptic currents and causing central sensitization.

A variety of animal model studies have shown that P2X3 receptors play an important role in the process of nociception. For example, P2X3 receptor knockout significantly reduces the pain response. P2X3 receptor antagonists have antineoplastic effects in various models of pain and inflammatory pain. In addition to its prominent role in nociception and acute and chronic pain, P2X3 receptors have also been shown to be involved in the pathological process of genitourinary system, gastrointestinal and respiratory diseases, especially overactive bladder and chronic cough. Therefore, P2X3 receptors play an important role in the pathological mechanisms of various diseases including pain, genitourinary system diseases, gastrointestinal diseases and respiratory diseases, and are ideal targets for the treatment of these diseases.

The P2X3 subunit not only forms a homotrimer, but also a heterotrimer with a P2X2 subunit. The P2X3 and P2X2 subunits are also expressed on the nerve fibers of the tongue, and receptors containing the P2X3 and/or P2X2 subunits participate in taste transmission (bitter, sweet, salty, umami and sour). Studies have shown that P2X3 homotrimers are mainly involved in mediating nociception, while P2X2/X3 heterotrimers are mainly involved in taste perception. Knockout animals lacking P2X2 and P2X3 subunits exhibit reduced taste and even loss of taste, while P2X3 subunit knockouts exhibit mild or no changes in phenotype (J. Physiol. 2015, 593, 1113–1125).

At present, the fastest research in the field of P2X3 receptor antagonists is the compound Gefapixant (AF-219) developed by Merck and Afferent (PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCE OF THE UNITED STATES OF AMERICA, 115(19), 4939-4944 .), the compound is a non-selective antagonist of P2X3 and P2X2/X3, and has shown significant efficacy in the clinical phase II study for chronic cough, but it also shows the side effects of taste interference. This side effect is mainly due to P2X2/3 heterotrimer blockade. Therefore, there is a continuing need for new or improved P2X3 antagonists for the development of new and more effective drugs to treat chronic cough or other diseases related to P2X3.

Summary of the invention

After painstaking research, the inventors designed and synthesized a series of substituted arylformamide compounds, and screened them for P2X3 activity. The research results showed that these compounds have outstanding P2X3 antagonistic activity and can be developed as therapeutic and / Or drugs to prevent diseases related to P2X3 activity.

Therefore, the object of the present invention is to provide a compound represented by the general formula (I) or its meso, racemate, enantiomer, diastereomer, or mixture form thereof, or Its medicinal salt,

in:

W ¹ , W ² , and W ³ are each independently selected from CR ⁶ or N;

A ¹ , A ² , A ³ , A ⁴ , and A ⁵ are each independently selected from C, N, O or S;

R ^{1 is} selected from -NR ^a R ^b , -NR ^a S(O) _m R ^b , -NR ^a S(O) _m NR ^a R ^b , -NR ^a S(O)(NR ^a )R ^b , -NR ^a S(O)(NR ^a )NR ^b , -NR ^a C(O)R ^b , -NR ^a C(O)NR ^a R ^b , -S(O) _m R ^a , -S(O) _m NR ^a R ^b , -S(O)(NR ^a )NR ^a R ^b , -OR ^a , -C(O)NR ^a R ^b , -P(O)R ^a R ^b , -(CR ^a R ^b )R ^b ;

Each R ^{2 is} independently selected from hydrogen, halogen, amino, nitro, cyano, hydroxyl, mercapto, oxo, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl , -C(O)R ^a , -O(O)CR ^a , -C(O)OR ^a , -C(O)NR ^a R ^b , -NHC(O)R ^a , -S(O) _m R ^{_{a, -S (O) m NR}} a R b, -NHS (O) m R a; wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl groups are optionally further substituted Selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxy, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, hetero One or more groups of aryl are substituted;

R ^{3 is} selected from aryl or heteroaryl; the aryl or heteroaryl is optionally further substituted with one or more groups selected from halogen, alkyl, haloalkyl, alkoxy, and haloalkoxy;

L is selected from -C(R ⁴ R ⁵ )-;

R ⁴ and R ⁵ are each independently selected from hydrogen, alkyl, alkoxy; wherein the alkyl and alkoxy are optionally further selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxy, One or more group substitutions of ester group, oxo group, alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

R ^{6 is} selected from hydrogen, halogen, hydroxyl, cyano, amino, carboxy, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein The alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group are optionally further selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, One or more group substitutions of ester group, oxo group, alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

R ^a and R ^b are each independently selected from hydrogen, halogen, hydroxyl, cyano, amino, carboxy, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, Heteroaryl groups, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups are optionally further selected from halogen, amino, nitro, cyano, One of hydroxyl, mercapto, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl or one of Multiple group substitutions;

Or R ^a and R ^b together with the atoms to which they are attached form a cycloalkyl or heterocyclic group, the cycloalkyl or heterocyclic group is optionally further selected from halogen, amino, nitro, cyano, oxo, One or more groups of hydroxyl, mercapto, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl replace;

m is 0, 1 or 2;

n is an integer from 0 to 3.

In a preferred embodiment of the present invention, the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, diastereomer, Or a mixture thereof, or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (II) or a meso, racemate, enantiomer, diastereomer, or In the form of a mixture, or a pharmaceutically acceptable salt thereof,

Wherein, A ¹ , A ² , A ³ , A ⁴ , A ⁵ , R ¹ , R ² , R ³ , L, and n are as defined in the general formula (I).

In another preferred embodiment of the present invention, the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, or diastereomer , Or its mixture form, or its pharmaceutically acceptable salt, wherein,

Group

Selected from pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isothiazolyl, isoxazolyl, 1,3,4-oxadiazolyl, 1,2,4 -Oxadiazolyl, 1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl, triazolyl or tetrazolyl, preferably pyrazolyl, thiazolyl, oxazolyl or 1, 3,4-oxadiazolyl, more preferably thiazolyl;

Said

Replaced by n R ^2;

R ² and n are as defined in the general formula (I).

In another preferred embodiment of the present invention, the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, or diastereomer , Or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R ³ is a C ₆ -C ₁₀ aryl group or a 5- to 10-membered heteroaryl group, preferably phenyl, pyridyl, pyrimidinyl, pyrazinyl, or pyridazine Group, more preferably pyrimidinyl, the aryl or heteroaryl group is optionally further selected from halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ One or more groups of -C _{6 haloalkoxy are substituted.}

In another preferred embodiment of the present invention, the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, or diastereomer , Or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein,

R ^{1 is} selected from -NR ^a R ^b , -NR ^a S(O) _m R ^b , -NR ^a S(O) _m NR ^a R ^b , -NR ^a S(O)(NR ^a )R ^b , -NR ^a S(O)(NR ^a )NR ^b , -NR ^a C(O)R ^b , -NR ^a C(O)NR ^a R ^b , -S(O) _m R ^a , -S(O) _m NR ^a R ^b , -S(O)(NR ^a )NR ^a R ^b , -OR ^a , -C(O)NR ^a R ^b , -P(O)R ^a R ^b , -(CR ^a R ^b )R ^b , preferably -NR ^a R ^b , -NR ^a S(O) ₂ R ^b , -NR ^a S(O)(NR ^a )R ^b , -NR ^a S(O) ₂ NR ^a R ^b , -NR ^a S(O)(NR ^a )NR ^b , -NR ^a C(O)R ^b , -NR ^a C(O)NR ^a R ^b , -S(O)R ^a , -SO ₂ R ^a , -S( O) ₂ NR ^a R ^b , -S(O)(NR ^a )NR ^a R ^b , -OR ^a , -P(O)R ^a R ^b ;

R ^a and R ^b are each independently selected from hydrogen, halogen, alkyl, alkoxy, cycloalkyl, and heterocyclyl, wherein the alkyl, alkoxy, cycloalkyl, and heterocyclyl are optionally further selected from Selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxy, ester, oxo, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, hetero One or more aryl groups are substituted;

Or R ^a and R ^b together with the atoms to which they are attached form a 5- to 8-membered heterocyclic group, the heterocyclic group is optionally further selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, One or more group substitutions of carboxyl group, ester group, alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

m is 0, 1, or 2.

In a preferred embodiment of the present invention, the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, or diastereomer , Or its mixture form, or its pharmaceutically acceptable salt, wherein,

R ^{1 is} selected from -NR ^a R ^b ;

R ^a is selected from hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 5-8 membered heterocyclyl particularly oxygen Etanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, wherein the C ₁ -C ₆ alkyl, C ₃ -C ₆ ring Alkyl and 5-8 membered heterocyclic groups are optionally further selected from halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy One or more groups of the group are substituted;

R ^{b is} selected from hydrogen or C ₁ -C ₆ alkyl.

In another preferred embodiment of the present invention, the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, or diastereomer Body, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein

R ^{1 is} selected from -NR ^a R ^b ;

R ^a and R ^b together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocyclic group, preferably tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, or 8-membered spiro heterocyclic group; The cyclic group is optionally further selected from one or more groups _{selected from halogen, C 1} -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C _{6 haloalkoxy} replace.

R ¹ is selected -S (O) R ^a, or -SO ₂ R ^a;

R ^a is selected from hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 5-8 membered heterocyclyl particularly oxygen Etanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, wherein the C ₁ -C ₆ alkyl, C ₃ -C ₆ ring Alkyl and 5-8 membered heterocyclic groups are optionally further selected from halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy One or more groups of the group are substituted.

R ^{1 is} selected from -S(O) ₂ NR ^a R ^b ;

R ^{b is} selected from hydrogen or C ₁ -C ₆ alkyl;

Alternatively, R ^a and R ^b together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocyclic group, preferably tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, or 8-membered spiro heterocyclic group; The heterocyclic group is optionally further selected from one or more of _{halogen, C 1} -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C _{6 haloalkoxy} Group substitution.

R ^{1 is} selected from -OR ^a ;

R ^{1 is} selected from -P(O)R ^a R ^b ;

R ^{b is} selected from hydrogen or C ₁ -C ₆ alkyl;

Or R ^a and R ^b together with the atoms to which they are attached form a 5- to 8-membered heterocyclic group, especially a phosphoryl group; the heterocyclic group is optionally further selected from halogen, C ₁ -C ₆ alkyl, One or more groups of C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, and C ₁ -C _{6 haloalkoxy are substituted.}

R ^{2 is} selected from hydrogen, halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₃ -C ₇ cycloalkyl, 5- to 7-membered heterocyclic group; preferably C ₁ -C ₆ alkyl ; Wherein the alkyl group, alkoxy group, cycloalkyl group, heterocyclic group are optionally further selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, alkyl, One or more groups of alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are substituted.

L is selected from -C(R ⁴ R ⁵ )-;

R ⁴ and R ⁵ are each independently selected from hydrogen, C ₁ -C ₆ alkyl, and C ₁ -C ₆ alkoxy.

Typical compounds of the present invention include but are not limited to:

The meso, racemate, enantiomer, diastereomer, or mixture form thereof, or a pharmaceutically acceptable salt thereof.

The present invention further provides a method for preparing the compound represented by the general formula (I) according to the present invention or its meso, racemate, enantiomer, diastereomer, or mixture thereof Form, its prodrug or its pharmaceutically acceptable salt method, which comprises the following steps:

In the presence of a condensing agent, the compound Ig and Ih are subjected to a condensation reaction under basic conditions to obtain a compound of general formula (I), wherein the basic condition is preferably DIPEA, and the condensing agent is preferably HATU;

Wherein, W ¹ , W ² , W ³ , A ¹ , A ² , A ³ , A ⁴ , A ⁵ , R ¹ , R ² , R ³ , L, and n are as defined in the general formula (I).

Another aspect of the present invention provides a pharmaceutical composition, which contains the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, or diastereomer Isomers, or mixtures thereof, prodrugs or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers.

The present invention further provides the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, and Use of a prodrug or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the prodrug in the preparation of a P2X3 antagonist.

The present invention further provides the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, and Use of a prodrug or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the prodrug in the preparation of a medicine for preventing and/or treating diseases related to P2X3 activity.

The present invention further provides the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, and A prodrug or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the prodrug, for use as a P2X3 antagonist.

The present invention further provides the compound represented by the general formula (I) according to the present invention or its mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, and A prodrug or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing it, for use as a medicine for the prevention and/or treatment of diseases related to P2X3 activity.

The present invention further provides a method for preventing and/or treating diseases associated with P2X3 activity, which comprises administering to a subject in need a preventive or therapeutically effective amount of the compound represented by the general formula (I) of the present invention The compound or its mesosome, racemate, enantiomer, diastereomer, or mixture thereof, its prodrug or its pharmaceutically acceptable salt, or a pharmaceutical composition containing it.

In a preferred embodiment of the present invention, the diseases related to P2X3 activity according to the present invention may be: respiratory diseases including chronic obstructive pulmonary disease (COPD), asthma, bronchospasm, pulmonary fibrosis, acute cough, Chronic cough, including chronic idiopathic and chronic refractory cough, genitourinary system, gastrointestinal, respiratory and pain-related diseases, gynecological diseases including dysmenorrhea (primary and secondary dysmenorrhea), dyspareunia, dysuria or Orchitis, endometriosis and adenomyosis, endometriosis-related pain, endometriosis-related symptoms, pelvic hypersensitivity, urinary tract disease state related to bladder outlet obstruction, urinary incontinence Symptoms such as decreased bladder capacity, increased urination frequency, urge incontinence, stress urinary incontinence or overreaction of the bladder, benign prostatic hypertrophy, prostatic hyperplasia, prostatitis, detrusor hyperreflexia, overactive bladder and overactive bladder Symptoms related to disease, especially frequent urination, nocturia, urgency or urge incontinence, pelvic hypersensitivity; urethritis, prostatitis, prostatic pain, cystitis, especially interstitial cystitis. Primary bladder hypersensitivity, epilepsy, partial and generalized seizures, gastrointestinal diseases including irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), biliary colic and other biliary diseases, renal colic, diarrhea Predominant IBS, gastroesophageal reflux, gastrointestinal dilatation, Crohn’s disease, neurodegenerative diseases such as Alzheimer’s disease, multiple sclerosis, Parkinson’s disease, cerebral ischemia and traumatic brain injury, Myocardial infarction, lipid disorders, pain-related diseases or conditions are selected from hyperalgesia, allodynia, functional bowel disease, gout, arthritis (such as osteoarthritis, rheumatoid arthritis and ankylosing spondylitis), Burning mouth syndrome, burns, migraine or cluster headache, nerve injury, post-traumatic injury (including fractures and sports injuries), neuritis, neuralgia, poisoning, ischemic injury, interstitial cystitis, cancer, trigeminal Neuralgia, small fiber neuropathy, diabetic neuropathy, chronic arthritis and related neuropathy, neuropathy caused by HIV and HIV treatment, pruritus, impaired wound healing and bone diseases such as joint degeneration.

According to conventional methods in the field to which the present invention belongs, the compound represented by the general formula (I) of the present invention can form a pharmaceutically acceptable acid addition salt with an acid. The acid includes inorganic acid and organic acid, particularly preferably hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, acetic acid, propionic acid, lactic acid , Trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc.

According to conventional methods in the field to which the present invention belongs, the compound represented by the general formula (I) of the present invention can form a pharmaceutically acceptable basic addition salt with a base. The base includes inorganic bases and organic bases. Acceptable organic bases include diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, etc., and acceptable inorganic bases include aluminum hydroxide, hydroxide Calcium, potassium hydroxide, sodium carbonate and sodium hydroxide, etc.

In addition, the present invention also includes prodrugs of the compounds represented by the general formula (I) of the present invention. The prodrugs of the present invention are derivatives of the compound represented by the general formula (I). They may have weak or even no activity by themselves, but after administration, under physiological conditions (for example, through metabolism, solvolysis) Or another way) is converted into the corresponding biologically active form.

The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, such as tablets, dragees, lozenges, water or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or Elixirs. Oral compositions can be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions can contain one or more ingredients selected from the group consisting of sweeteners, flavoring agents, coloring agents and preservatives, To provide pleasing and delicious medicinal preparations. The tablet contains the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for the preparation of tablets for mixing. These excipients can be inert excipients, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as microcrystalline cellulose, croscarmellose sodium, corn Starch or alginic acid; binders, such as starch, gelatin, polyvinylpyrrolidone, or gum arabic; and lubricants, such as magnesium stearate, stearic acid, or talc. These tablets may be uncoated or may be coated by known techniques that mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release effect over a longer period of time. For example, water-soluble taste-masking substances such as hydroxypropyl methylcellulose or hydroxypropyl cellulose, or extended time substances such as ethyl cellulose, cellulose acetate butyrate may be used.

Hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin can also be used, or in which the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oil solvent such as peanut oil, liquid paraffin or olive oil. Soft gelatin capsules provide oral preparations.

Aqueous suspensions contain the active substance and excipients suitable for the preparation of aqueous suspensions for mixing. Such excipients are suspending agents, such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinylpyrrolidone and gum arabic; dispersing or wetting agents, which can be natural Produced phospholipids such as lecithin, or condensation products of alkylene oxides and fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide and long-chain fatty alcohols, such as seventeen-carbon ethyleneoxy whale Heptadecaethyleneoxy cetanol, or condensation products of ethylene oxide and partial esters derived from fatty acids and hexitols, such as polyethylene oxide sorbitol monooleate, or ethylene oxide with fatty acids and hexitols Condensation products of anhydride-derived partial esters, such as polyethylene oxide sorbitan monooleate. The aqueous suspension may also contain one or more preservatives such as ethyl paraben or n-propyl paraben, one or more coloring agents, one or more flavoring agents and one or more sweeteners. Flavoring agents such as sucrose, saccharin or aspartame.

Oil suspensions can be formulated by suspending the active ingredients in vegetable oils such as peanut oil, olive oil, sesame oil or coconut oil, or mineral oils such as liquid paraffin. Oil suspensions may contain thickeners such as beeswax, hard paraffin or cetyl alcohol. The above-mentioned sweeteners and flavoring agents can be added to provide a palatable preparation. These compositions can be preserved by adding antioxidants such as butylated hydroxyanisole or alpha-tocopherol.

By adding water, dispersible powders and granules suitable for preparing aqueous suspensions can provide the active ingredient and a dispersing or wetting agent for mixing, a suspending agent or one or more preservatives. Suitable dispersing or wetting agents and suspending agents are as described above. Other excipients such as sweeteners, flavoring agents and coloring agents may also be added. These compositions are preserved by adding antioxidants such as ascorbic acid.

The pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion. The oil phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin or a mixture thereof. Suitable emulsifiers can be naturally occurring phospholipids, such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation of the partial esters and ethylene oxide Products such as polyethylene oxide sorbitol monooleate. The emulsion may also contain sweetening agents, flavoring agents, preservatives and antioxidants. Syrups and elixirs formulated with sweetening agents such as glycerin, propylene glycol, sorbitol or sucrose. Such preparations may also contain a demulcent, a preservative, a coloring agent and an antioxidant.

The pharmaceutical composition of the present invention may be in the form of a sterile injectable aqueous solution. Acceptable solvents and solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. Then the oil solution is added to the mixture of water and glycerin to form a microemulsion. The injection or microemulsion can be injected into the patient's bloodstream by local large-scale injection. Alternatively, it is best to administer the solution and microemulsion in a manner that maintains a constant circulating concentration of the compound of the present invention. To maintain this constant concentration, a continuous intravenous delivery device can be used.

The pharmaceutical composition of the present invention may be in the form of a sterile injection water or oil suspension for intramuscular and subcutaneous administration. The suspension can be formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents mentioned above. The sterile injection preparation may also be a sterile injection solution or suspension prepared in a non-toxic parenterally acceptable diluent or solvent, for example, a solution prepared in 1,3-butanediol. In addition, sterile fixed oil can be conveniently used as a solvent or suspension medium. For this purpose, any blended fixed oil including synthetic mono- or diglycerides can be used. In addition, fatty acids such as oleic acid can also be used to prepare injections.

The compounds of the present invention can be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid in the rectum and thus will melt in the rectum to release the drug. Such substances include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights, and mixtures of fatty acid esters of polyethylene glycol.

Those skilled in the art know that the dosage of drugs depends on many factors, including but not limited to the following factors: the activity of the specific compound used, the patient’s age, the patient’s weight, the patient’s health, the patient’s clothing, and the patient’s The diet, time of administration, mode of administration, rate of excretion, combination of drugs, etc. In addition, the best treatment method, such as the treatment mode, the daily dosage of the compound of the general formula, or the type of pharmaceutically acceptable salt can be verified according to the traditional treatment plan.

The present invention may contain a compound represented by general formula (I), and a pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient, mixed with a pharmaceutically acceptable carrier or excipient to prepare a composition, and Prepared into a clinically acceptable dosage form. The derivatives of the present invention can be used in combination with other active ingredients as long as they do not produce other adverse effects, such as allergic reactions and the like. The compound of the present invention can be used as the sole active ingredient, and can also be used in combination with other drugs for the treatment of diseases related to P2X3 activity. Combination therapy is achieved by administering the individual therapeutic components simultaneously, separately or sequentially.

Detailed description of the invention

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably containing 1 to 6 carbons Atom of the alkyl group. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1 ,2-Dimethylpropyl, 2,2-Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2- Methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3 -Dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2 -Methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethyl Pentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2 ,5-Dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethyl 2-methylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethyl Base hexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers. More preferred are lower alkyl groups containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and sec-butyl. Group, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl Butyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl Group, 2,3-dimethylbutyl, etc. Alkyl groups may be substituted or unsubstituted. When substituted, substituents may be substituted at any available attachment point. The substituents are preferably one or more of the following groups, which are independently selected from alkanes Group, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkane An oxy group, a heterocycloalkoxy group, a cycloalkylthio group, a heterocycloalkylthio group, an oxo group, a carboxyl group or a carboxylate group.

The term "alkenyl" refers to an alkyl group as defined above composed of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3 -Butenyl etc. Alkenyl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycle Alkylthio.

The term "alkynyl" refers to an alkyl group as defined above composed of at least two carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, propynyl, butynyl, and the like. The alkynyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycle Alkylthio.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent. The cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 Carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene Groups, cyclooctyl, etc.; polycyclic cycloalkyls include spiro, fused, and bridged cycloalkyls.

The term "spirocycloalkyl" refers to a polycyclic group that shares one carbon atom (called a spiro atom) between 5- to 20-membered monocyclic rings, which may contain one or more double bonds, but none of the rings have complete conjugate Π electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the ring and the ring, the spirocycloalkyl group is classified into a single spirocycloalkyl group, a bispirocycloalkyl group or a polyspirocycloalkyl group, preferably a single spirocycloalkyl group and a bispirocycloalkyl group. More preferably, it is a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, or 5-membered/6-membered monospirocycloalkyl. Non-limiting examples of spirocycloalkyl groups include:

The term "fused cycloalkyl" refers to a 5- to 20-membered all-carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more of the rings may contain one or Multiple double bonds, but none of the rings have a fully conjugated π-electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic condensed cycloalkyls, preferably bicyclic or tricyclic, and more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic alkyl. Non-limiting examples of fused cycloalkyl groups include:

The term "bridged cycloalkyl" refers to a 5- to 20-membered, all-carbon polycyclic group with any two rings sharing two carbon atoms that are not directly connected. It may contain one or more double bonds, but no ring has a complete Conjugated π electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyls, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl groups include:

The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, non-limiting examples include indanyl, tetrahydronaphthalene Group, benzocycloheptanyl group, etc. Cycloalkyl groups may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , Heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) _m (where m is an integer of 0 to 2) heteroatoms, but does not include the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably contains 5 to 7 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidine Group, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably 1, 2, 5-oxadiazolyl, pyranyl or morpholinyl. Polycyclic heterocyclic groups include spiro, fused, and bridged heterocyclic groups.

The term "spiroheterocyclic group" refers to a polycyclic heterocyclic group that shares one atom (called a spiro atom) between 5- to 20-membered monocyclic rings, in which one or more ring atoms are selected from nitrogen, oxygen or S(O ) _{Heteroatoms of m} (where m is an integer of 0 to 2), and the remaining ring atoms are carbon. It can contain one or more double bonds, but none of the rings have a fully conjugated π-electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the ring and the ring, the spiro heterocyclic group is classified into a single spiro heterocyclic group, a dispiro heterocyclic group or a polyspiro heterocyclic group, preferably a single spiro heterocyclic group and a dispiro heterocyclic group. More preferably, it is a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, or 5-membered/6-membered monospiro heterocyclic group. Non-limiting examples of spiroheterocyclic groups include:

The term "fused heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system. One or more rings may contain one or more Double bond, but none of the rings have a fully conjugated π-electron system, where one or more of the ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (where m is an integer from 0 to 2), and the rest of the ring The atom is carbon. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic condensed heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic condensed heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

The term "bridged heterocyclic group" refers to a 5- to 14-membered polycyclic heterocyclic group with any two rings sharing two atoms that are not directly connected. It may contain one or more double bonds, but none of the rings has a complete common A conjugated π-electron system in which one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O) _m (where m is an integer of 0 to 2), and the remaining ring atoms are carbon. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

The heterocyclic ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, non-limiting examples of which include:

Wait.

The heterocyclic group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , Heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "aryl" refers to a 6 to 14-membered all-carbon monocyclic or fused polycyclic (that is, rings sharing adjacent pairs of carbon atoms) with a conjugated π-electron system, preferably 6 to 10 members, such as benzene Base and naphthyl. Phenyl is more preferred. The aryl ring may be fused on a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples of which include:

The aryl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycle Alkylthio, carboxy, or carboxylate.

The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, where the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl groups are preferably 5- to 10-membered, containing 1 to 3 heteroatoms; more preferably 5-membered or 6-membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furyl, thienyl, thiazolyl, pyridine Azolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, thiazolyl, pyrazolyl or pyrimidinyl, thiazolyl; more preferably Pyrazolyl or thiazolyl. The heteroaryl ring may be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

Heteroaryl groups may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , Heterocycloalkylthio, carboxyl or carboxylate.

The term "alkoxy" refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), where the definition of alkyl is as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. The alkoxy group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , Heterocycloalkylthio, carboxyl or carboxylate.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, where the alkyl group is as defined above.

The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, where alkoxy is as defined above.

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, where the alkyl group is as defined above.

The term "hydroxy" refers to the -OH group.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "amino" refers to -NH ₂ .

The term "cyano" refers to -CN.

The term "nitro" refers to -NO ₂ .

The term "oxo" refers to =O.

The term "carboxy" refers to -C(O)OH.

The term "mercapto" refers to -SH.

The term "ester group" refers to -C(O)O (alkyl) or -C(O)O (cycloalkyl), where alkyl and cycloalkyl are as defined above.

The term "acyl" refers to a compound containing a -C(O)R group, where R is an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group, or a heteroaryl group.

The term "sulfonic acid" refers to -S(O) ₂ OH.

The term "sulfonate group" refers to -S(O) ₂ O (alkyl) or -S(O) ₂ O (cycloalkyl), where alkyl and cycloalkyl are as defined above.

The term "sulfonyl" refers to _{a compound of the -S(O) 2} R group, where R is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

The term "aminoacyl" refers to -C(O)-NRR', wherein R and R'are each independently hydrogen, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl.

The term "aminosulfonyl" or "sulfonylamino" refers to -S(O) ₂ -NRR', wherein R and R'are each independently hydrogen, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl base.

"Optional" or "optionally" means that the event or environment described later can but does not have to occur, and the description includes occasions where the event or environment occurs or does not occur. For example, "heterocyclic group optionally substituted by an alkyl group" means that an alkyl group may but does not have to be present. The description includes the case where the heterocyclic group is substituted by an alkyl group and the case where the heterocyclic group is not substituted by an alkyl group. .

"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms independently of each other substituted by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without too much effort. For example, an amino group or a hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiological/pharmaceutically acceptable carriers And excipients. The purpose of the pharmaceutical composition is to promote the administration to the organism, facilitate the absorption of the active ingredients and then exert the biological activity.

"Pharmaceutically acceptable salt" refers to the salt of the compound of the present invention, which is safe and effective when used in mammals, and has due biological activity.

Synthetic method of the compound of the present invention

In order to accomplish the purpose of the present invention, the present invention adopts the following technical solutions.

The compound represented by the general formula (I) of the present invention or its mesosome, racemate, enantiomer, diastereomer, or mixture form thereof, or a pharmaceutically acceptable salt thereof can be obtained by Scheme preparation, the specific preparation method is as follows.

(1) Scheme 1: When R ¹ is -NR ^a R ^b , according to the method of Scheme 1, using compound Ia as the starting material, the compound of general formula (I) is obtained.

plan 1

Synthesis of Scheme 1:

Step 1: Reacting compound Ia with Ra (O) under acidic conditions and the presence of ^a reducing agent to obtain compound Ib, wherein the acidic reagent is preferably acetic acid, and the reducing agent is preferably sodium cyanoborohydride;

Step 2: Under alkaline conditions, reacting compound Ib with R ^b I to obtain compound Ic, wherein the alkaline reagent is preferably NaH;

Step 3: In the presence of high temperature, alkalinity and catalyst, compound Ic is reacted with pinacol diboronic acid to obtain compound Id, wherein the high temperature condition is preferably 100°C, the alkaline reagent is preferably potassium acetate, and the catalyst is preferably Pd(dppf) Cl ₂ catalyst;

Step 4: In the presence of high temperature, alkalinity and catalyst, compound Id is reacted with compound Ie to obtain compound If, wherein the high temperature condition is preferably 90°C, the alkaline reagent is preferably potassium carbonate, and the catalyst is preferably Pd(dppf)Cl ₂ catalyst;

Step 5: Under alkaline conditions, hydrolyze compound If to obtain compound Ig, wherein the alkaline reagent is preferably lithium hydroxide;

Step 6: In the presence of a condensing agent, the compound Ig and Ih are subjected to a condensation reaction under basic conditions to obtain a compound of general formula (I), wherein the basic condition is preferably DIPEA, and the condensing agent is preferably HATU.

(2) Scheme 2: When R ¹ is -SO ₂ R ^a, according to the method of Scheme 2, the compound Ii as starting materials, to give compounds of general formula (I).

Scenario 2

Synthesis of Scheme 2:

Step 1: The compound Ii is reacted with (4-methoxyphenyl)methanethiol in the presence of high temperature, basicity and a catalyst to obtain compound Ij, wherein the high temperature condition is preferably 100°C, the alkaline reagent is preferably DIPEA, and the catalyst is preferably Pd ₂ (dba) ₃ and Xantphos;

Step 2: In the presence of high temperature, alkalinity and catalyst, compound Ij is reacted with pinacol diboronic acid ester to obtain compound Ik, wherein the high temperature condition is preferably 100°C, the alkaline reagent is preferably potassium acetate, and the catalyst is preferably Pd(dppf) Cl ₂ ;

Step 3: In the presence of high temperature, alkalinity and catalyst, compound Ik is reacted with compound Ie to obtain compound Il, wherein the high temperature condition is preferably 90°C, the alkaline reagent is preferably potassium carbonate, and the catalyst is preferably Pd(dppf)Cl ₂ ;

Step 4: In the presence of a low temperature and an oxidizing agent, the compound 11 is reacted to obtain the compound Im, wherein the low temperature condition is preferably -15°C, and the oxidizing agent is preferably 1,3-dichloro-5,5-dimethylhydantoin (DCDMH);

Step 5: under basic conditions, with Im HR ^a compound obtained by reacting a compound of In, wherein the alkaline reagent is preferably sodium carbonate;

Step 6: Under alkaline conditions, hydrolyze compound In to obtain compound Io, wherein the alkaline reagent is preferably lithium hydroxide;

Step 7: In the presence of a condensing agent, the compound Io and Ih are subjected to a condensation reaction under basic conditions to obtain a compound of general formula (I), wherein the basic condition is preferably DIPEA, and the condensing agent is preferably HATU.

(3) Scheme 3: When R ¹ is -S(O) ₂ NR ^a R ^b , according to the method of Scheme 3, using compound Im as the starting material, the compound of general formula (I) is obtained.

Scheme 3

Synthesis of Scheme 3:

Step 1: Under alkaline conditions, reacting compound Im with HNR ^a R ^b to obtain compound Ip, wherein the alkaline reagent is preferably sodium carbonate;

Step 2: Under alkaline conditions, the compound Ip is hydrolyzed to obtain the compound Iq, wherein the alkaline reagent is preferably lithium hydroxide;

Step 3: In the presence of a condensing agent, the compound Iq and Ih are subjected to a condensation reaction under basic conditions to obtain a compound of general formula (I), wherein the basic condition is preferably DIPEA, and the condensing agent is preferably HATU.

(4) Scheme 4: When R ¹ is -P(O)R ^a R ^b , according to the method of Scheme 4, using compound Ir as the starting material, the compound of general formula (I) is obtained.

Scheme 4

Synthesis of Scheme 4:

Step 1: In the presence of high temperature, alkalinity and catalyst, compound Ir is reacted with HP(O)R ^a R ^b to obtain compound Is, wherein the high temperature condition is preferably 65°C, the alkaline reagent is preferably potassium phosphate, and the catalyst is preferably Pd(OAc ) ₂ and Xantphos;

Step 2: In the presence of high temperature, alkalinity and catalyst, compound Is is reacted with diboronic acid pinacol ester to obtain compound It, wherein the high temperature condition is preferably 100°C, the alkaline reagent is preferably potassium acetate, and the catalyst is preferably Pd(dppf) Cl ₂ ;

Step 3: In the presence of high temperature, alkalinity and catalyst, compound It and Ie are reacted to obtain compound Iu, wherein the high temperature condition is preferably 90°C, the alkaline reagent is preferably potassium carbonate, and the catalyst is preferably Pd(dppf)Cl ₂ ;

Step 4: Under alkaline conditions, hydrolyze compound Iu to obtain compound Iv, wherein the alkaline reagent is preferably lithium hydroxide;

Step 5: In the presence of a condensing agent, the compound Iv and Ih are subjected to a condensation reaction under basic conditions to obtain a compound of general formula (I), wherein the basic condition is preferably DIPEA, and the condensing agent is preferably HATU.

Wherein, W ¹ , W ² , W ³ , A ¹ , A ² , A ³ , A ⁴ , A ⁵ , R ¹ , R ² , R ³ , and L are as defined in the general formula (I).

Detailed ways

The present invention is further described below in conjunction with examples, but these examples do not limit the scope of the present invention.

The structure of the compound is determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS). NMR shifts are given in units of ^{10 -6 (ppm).} NMR is measured by Brukerdps300 nuclear magnetometer, the solvent is deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and the internal standard is tetramethyl Silane (TMS).

1100Series LC/MSD Trap (ESI) mass spectrometer (manufacturer: Agilent) is used for MS measurement.

GC-MS measurement uses GCMS-QP2010SE.

The lc3000 high performance liquid chromatograph and lc6000 high performance liquid chromatograph (manufacturer: Chuangxin Tongheng) are used for the preparation of the liquid phase. The chromatographic column is Daisogel C18 10μm 60A (20mm×250mm).

HPLC determination uses Shimadzu LC-20AD high pressure liquid chromatograph (Agilent TC-C18 250×4.6mm 5μm column) and Shimadzu LC-2010AHT high pressure liquid chromatograph (Phenomenex C18 250×4.6mm 5μm column).

The thin layer chromatography silica gel plate uses Qingdao Ocean Chemical GF254 silica gel plate, the size of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm～0.2mm, and the size of thin layer chromatography separation and purification products is 0.4mm～0.5 mm.

Column chromatography generally uses Qingdao Marine silica gel 100-200 mesh and 200-300 mesh silica gel as the carrier.

The known starting materials of the present invention can be synthesized by or according to methods known in the art, or can be purchased from the network shopping mall, Beijing Coupling, Sigma, Bailingwei, Yi Shiming, Shanghai Shuya, Inokia, Nanjing Yaoshi, Anaiji Chemical and other companies.

There is no special description in the examples, and the reaction can all be carried out under an argon atmosphere or a nitrogen atmosphere.

The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1L.

The microwave reaction uses CEM Discover SP type microwave reactor.

There is no special description in the examples, and the solution refers to an aqueous solution.

There are no special instructions in the examples, and the reaction temperature is room temperature, which is 20°C to 30°C.

The monitoring of the reaction progress in the examples adopts thin-layer chromatography (TLC). The developing reagent system used in the reaction includes: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether And ethyl acetate system, D: acetone, the volume ratio of the solvent is adjusted according to the polarity of the compound.

The eluent system of column chromatography and the developing solvent system of thin layer chromatography used to purify compounds include: A: dichloromethane and methanol system, B: petroleum ether, ethyl acetate and dichloromethane system, C: petroleum In the ether and ethyl acetate system, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of basic or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.

Unless otherwise defined, all professional and scientific terms used in the text have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to the content described can be applied to the method of the present invention.

Example

Example 1: (R)-3-(Methyl(oxetan-3-yl)amino)-5-(5-methylthiazol-2-yl)-N-(1-(2-( Preparation of (trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (1)

Step 1: Synthesis of methyl 3-bromo-5-(oxetan-3-ylamino)benzoate (1b)

Mix 3-amino-5-bromobenzoic acid methyl ester (2.00g, 8.69mmol), oxetane-3-one (1.25g, 17.38mmol), acetic acid (1.58g, 26.07mmol), absolute ethanol ( 30mL) was added to the reaction flask, heated to 60°C and stirred for 5 hours, then sodium cyanoborohydride (1.58g, 26.07mmol) was added, and stirring was continued at 60°C for 14 hours. After the reaction, the reaction solution was filtered, and saturated sodium bicarbonate solution was added to the filtrate to adjust the pH to 8-9, and then 50 mL of dichloromethane and 50 mL of water were added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 2.34 g of the title product as a pale yellow solid with a yield of 93%.

Step 2: Synthesis of methyl 3-bromo-5-(methyl(oxetan-3-yl)amino)benzoate (1c)

Add methyl 3-bromo-5-(oxetan-3-ylamino)benzoate (2.34g, 8.18mmol) and DMF (30ml) into the reaction flask, and then slowly add NaH (0.78g, 32.72mmol) ), stirring at room temperature for 1 hour, and then adding methyl iodide (2.32 g, 16.36 mmol), and continuing to react at room temperature for 14 hours. After the completion of the reaction, 10 mL of water was added dropwise to the reaction solution to quench. The reaction solution was concentrated under reduced pressure, and 60 mL of dichloromethane and 60 mL of water were added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 1.30 g of the title product as a white solid with a yield of 53%.

Step 3: 3-(Methyl(oxetan-3-yl)amino)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane Synthesis of Alk-2-yl) Methyl Benzoate (1d)

Combine 3-bromo-5-(methyl(oxetan-3-yl)amino)methyl benzoate (1.15g, 32.72mmol), BPD (pinacol diboronate) (1.46g, 5.75 mmol), potassium acetate (0.75g, 7.66mmol), 1,4-dioxane (25ml), Pd(dppf)Cl ₂ (0.14g, 0.19mmol) were added to the reaction flask, and under nitrogen protection, the temperature was raised to 100 Stir at °C for 20 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was extracted with 50 mL of ethyl acetate and 50 mL of water. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 0.96 g of the title product as a white solid with a yield of 63%.

Step 4: Synthesis of methyl 3-(methyl(oxetan-3-yl)amino)-5-(5-methylthiazol-2-yl)benzoate (1e)

Add 3-(methyl(oxetan-3-yl)amino)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- Methyl 2-yl)benzoate (0.96g, 2.76mmol), 2-bromo-5-methylthiazole (0.59g, 3.32mmol), potassium carbonate (0.92g, 6.65mmol), water (5mL), tetrahydrofuran ( 25mL), Pd(dppf)Cl ₂ (0.30g, 0.86mmol) were added to the reaction flask, and under the protection of nitrogen, the temperature was raised to 90°C and stirred for 16 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was extracted with 50 mL of ethyl acetate and 50 mL of water. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 0.8 g of the title product as a light green liquid with a yield of 90%.

Step 5: Synthesis of 3-(methyl(oxetan-3-yl)amino)-5-(5-methylthiazol-2-yl)benzoic acid (1f)

Combine 3-(methyl(oxetan-3-yl)amino)-5-(5-methylthiazol-2-yl)methyl benzoate (0.70g, 2.20mmol), 1N lithium hydroxide solution (10 mL), tetrahydrofuran (20 mL), methanol (20 mL) were added to the reaction flask, and stirred at room temperature for 14 hours. After the reaction, the reaction solution was concentrated under reduced pressure, 1N hydrochloric acid solution was added to adjust the pH to 3-4, and 50 mL ethyl acetate and 50 mL water were added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain 0.16 g of the title product as a yellow solid with a yield of 24%.

Step 6: (R)-3-(Methyl(oxetan-3-yl)amino)-5-(5-methylthiazol-2-yl)-N-(1-(2-(tri Synthesis of (fluoromethyl)pyrimidin-5-yl)ethyl)benzamide (compound 1)

Add 3-(methyl(oxetan-3-yl)amino)-5-(5-methylthiazol-2-yl)benzoic acid (80mg, 0.26mmol), (R)-1-(2 -(Trifluoromethyl)pyrimidin-5-yl)ethyl-1-amine (prepared according to document WO2010111059) (66mg, 0.29mmol), DIPEA (diisopropylethylamine) (136mg, 1.05mmol), HATU ( 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate) (140g, 0.37mmol) and DMF (10mL) were added to the reaction flask and stirred at room temperature 2 hours. After the reaction, 30 mL ethyl acetate and 30 mL water were added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was prepared and purified by preparative liquid chromatography (column: Hedea ODS-2 C18, 300mm*250mm, 10μm; eluent: acetonitrile/water=0%-100%) to obtain 55mg of the title product as a yellow solid, yield 44%.

LC-MS: m/z 478.41 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 1.11 (m, 3H), 1.61 (m, 3H), 2.99 (m, 3H), 4.63 (m, 2H), 4.82 (m, 2H), 4.87 ( m, 1H), 5.29 (m, 1H), 7.23 (m, 2H), 7.62 (m, 1H), 7.70 (m, 1H), 9.10 (m, 3H).

Example 2: (R)-3-(5-Methylthiazol-2-yl)-5-(pyrrolidin-1-ylsulfonyl)-N-(1-(2-(trifluoromethyl)pyrimidine -5-yl) ethyl) benzamide (2) preparation

Step 1: Synthesis of methyl 3-bromo-5-((4-methoxybenzyl)thio)benzoate (2b)

3,5-Dibromobenzoic acid methyl ester (5.0g, 17.0mmol), (4-methoxyphenyl) methyl mercaptan (2.09g, 13.6mmol), Pd ₂ (dba) ₃ (1.95g, 3.4 mmol), Xantphos (4,5-bis(diphenylphosphine)-9,9-dimethylxanthene) (1.95g, 3.4mmol), dioxane (50mL), DIPEA (4.4g, 34mmol) ) Was added to the reaction flask and stirred at 100°C for 16 hours under the protection of nitrogen. After the reaction, the reaction solution was filtered, and the filtrate was concentrated to dryness. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 4.16 g of the title product as a yellow oil, yield: 66.8%.

Step 2: 3-((4-Methoxybenzyl)thio)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2 -Yl) the synthesis of methyl benzoate (2c)

Combine 3-bromo-5-((4-methoxybenzyl)thio)methyl benzoate (4.16g, 11.4mmol), BPD (4.33g, 17.0mmol), potassium acetate (2.23g, 22.8mmol) , Pd(dppf)Cl ₂ (0.42g, 0.57mmol) and DMF (40mL) were added to the reaction flask and stirred at 110°C for 3 hours. After the reaction, the reaction solution was filtered, and the filtrate was concentrated to dryness. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 5.2 g of the title product as a yellow oil, yield: 110%.

Step 3: Synthesis of methyl 3-((4-methoxybenzyl)thio)-5-(5-methylthiazol-2-yl)benzoate (2d)

Add 3-((4-methoxybenzyl)thio)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl ) Methyl benzoate (5.2g, 12.47mmol), 2-bromo-5-methylthiazole (2.66g, 14.96mmol), potassium carbonate (4.13g, 29.93mmol), Pd(dppf)Cl ₂ (1.37g, 1.87mmol), THF (200mL), and water (30mL) were added to the reaction flask, and stirred at 90°C for 16 hours under nitrogen protection. After the completion of the reaction, the reaction solution was filtered, and 100 mL of ethyl acetate and 100 mL of water were added to the filtrate for extraction. The organic phase was washed once with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain 2.6 g of the title product as a yellow oil, yield: 54.1%.

Step 4: Synthesis of methyl 3-(chlorosulfonyl)-5-(5-methylthiazol-2-yl)benzoate (2e)

Combine 3-((4-methoxybenzyl)sulfanyl)-5-(5-methylthiazol-2-yl)benzoic acid methyl ester (2.4g, 6.2mmol), acetonitrile (80mL), water (2mL ), acetic acid (1mL) were added to the reaction flask, cooled to -15℃, DCDMH (1,3-dichloro-5,5-dimethylhydantoin) (1.7g, 8.7mmol) was added to the reaction flask , Stir at -15°C for 4 hours. After the completion of the reaction, 80 mL of dichloromethane and 80 mL of water were added to the reaction solution for extraction, the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 1:1) to obtain 1.46 g of the title product as a white solid, yield: 70.8%.

Step 5: Synthesis of methyl 3-(5-methylthiazol-2-yl)-5-(pyrrolidin-1-ylsulfonyl)benzoate (2f)

Mix 3-(chlorosulfonyl)-5-(5-methylthiazol-2-yl) methyl benzoate (1.3g, 3.92mmol), THF (20mL), water (20mL), sodium carbonate (1.25g, 11.78mmol) and tetrahydropyrrole (0.84g, 11.78mmol) were added to the reaction flask and stirred at room temperature for 16 hours. After the reaction, the reaction solution was filtered, the filter cake was washed twice with water, and the filter cake was dried to obtain 1.4 g of the title product as a white solid. Yield: 97.4%.

Step 6: Synthesis of 3-(5-methylthiazol-2-yl)-5-(pyrrolidin-1-ylsulfonyl)benzoic acid (2g)

Combine 3-(5-methylthiazol-2-yl)-5-(pyrrolidin-1-ylsulfonyl) methyl benzoate (1.4g, 3.8mmol), THF (20mL), methanol (20mL), 1N Lithium hydroxide (20 mL) was added to the reaction flask and stirred at room temperature for 16 hours. After the reaction, the reaction solution was adjusted to pH 3-4 with 1N hydrochloric acid, filtered, the filter cake was washed twice with water, and the filter cake was dried to obtain 1.3 g of the title product as a white solid. Yield: 96.5%.

Step 7: (R)-3-(5-Methylthiazol-2-yl)-5-(pyrrolidin-1-ylsulfonyl)-N-(1-(2-(trifluoromethyl)pyrimidine- Synthesis of 5-yl)ethyl)benzamide (compound 2)

Combine 3-(5-methylthiazol-2-yl)-5-(pyrrolidin-1-ylsulfonyl)benzoic acid (140mg, 0.40mmol), DMF (10mL), HATU (213mg, 0.56mmol), DIPEA (206mg, 1.60mmol), (R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethan-1-amine (100mg, 0.44mmol) was added to the reaction flask and stirred at room temperature for 16 hours . After the reaction, the reaction solution was poured into 100 mL ice water, stirred for 10 minutes, filtered, and the filter cake was passed through preparative liquid chromatography (column: Hedea ODS-2 C18, 300mm*250mm, 10μm; eluent: 0%- 100% acetonitrile: aqueous solution) to obtain 107 mg of the title product as a white solid, yield: 51.2%.

LC-MS: m/z 526.29 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 1.67 (m, 8H), 2.55 (m, 3H), 3.19 (m, 3H), 5.31 (m, 1H), 7.74 (m, 1H), 8.33 ( m, 2H), 8.60 (m, 1H), 9.16 (s, 2H), 9.50 (m, 1H).

Example 3: (R)-3-(5-Methylthiazol-2-yl)-5-sulfamoyl-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl ) Preparation of benzamide (3)

The preparation method was the same as in Example 2, except that ammonia water was used instead of tetrahydropyrrole to obtain the title compound 3.

LC-MS: m/z 472.4 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 1.63 (m, 3H), 2.50 (s, 3H), 5.34 (m, 1H), 7.60 (m, 2H), 7.73 (s, 1H), 8.35 ( s, 1H), 8.46 (s, 1H), 8.53 (s, 1H), 9.14 (s, 2H), 9.47 (m, 1H).

Example 4: (R)-3-((4-methylpiperazin-1-yl)sulfonyl)-5-(5-methylthiazol-2-yl)-N-(1-(2-( Preparation of (trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (4)

The preparation method was the same as in Example 2, except that N-methylpiperazine was used instead of tetrahydropyrrole to obtain the title compound 4.

LC-MS: m/z 555.4 [M+H] ⁺ .

¹ H NMR(300MHz,DMSO-d ₆ )δ1.73(m,3H), 2.12(m,3H), 2.36(m,4H), 2.49(m,3H), 2.06(m,4H), 5.92( m, 1H), 7.74 (m, 1H), 8.22 (m, 2H), 8.01 (m, 1H), 8.14 (s, 2H), 9.04 (m, 1H).

Example 5: (R)-3-(N-cyclopropylsulfamoyl)-5-(5-methylthiazol-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidine -5-yl) ethyl) benzamide (5) preparation

The preparation method was the same as in Example 2, except that cyclopropylamine was used instead of tetrahydropyrrole to obtain the title compound 5.

LC-MS: m/z 512.4 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 0.40 (m, 2H), 0.50 (m, 2H), 1.63 (m, 3H), 2.21 (m, 1H), 2.55 (s, 3H), 5.34 ( m, 1H), 7.74 (m, 1H), 8.18 (s, 1H), 8.34 (s, 1H), 8.43 (s, 1H), 8.58 (s, 1H), 9.15 (s, 2H), 9.49 (m ,1H).

Example 6: (R)-3-(Dimethylphosphono)-5-(5-methylthiazol-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidine-5- (Yl) ethyl) benzamide (6) preparation

Step 1: Synthesis of methyl 3-bromo-5-(dimethylphosphono)benzoate (6b)

Mix 3-bromo-5-iodobenzoic acid methyl ester (6a) (2.00g, 5.87mmol), dimethylphosphine oxide (0.69g, 8.80mmol), potassium phosphate (1.50g, 7.04mmol), 1,4- Dioxane (15 mL), Xantphos (0.2 g, 0.35 mmol), and Pd(OAc) ₂ (65 mg, 0.29 mmol) were added to the reaction flask, and under the protection of nitrogen, the temperature was raised to 65° C. and stirred for 2 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane: methanol = 20:1) to obtain 0.8 g of the title product as an orange solid with a yield of 47 %.

Step 2: Methyl 3-(dimethylphosphono)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate Synthesis of ester (6c)

Mix 3-bromo-5-(dimethylphosphono) methyl benzoate (0.5g, 1.72mmol), BPD (0.87g, 3.44mmol), potassium acetate (0.5g, 5.16mmol), 1,4-bis Oxane (20 mL) and Pd(dppf)Cl ₂ (63 mg, 0.086 mmol) were added to the reaction flask, and under the protection of nitrogen, the temperature was raised to 100° C. and the mixture was stirred for 16 hours. After the reaction, it was filtered, and the filtrate was concentrated under reduced pressure to obtain 1.3 g of the title product as a crude black solid, which was used directly in the next step.

Step 3: Synthesis of methyl 3-(dimethylphosphono)-5-(5-methylthiazol-2-yl)benzoate (6d)

Methyl 3-(dimethylphosphono)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate ( 1.3g, 3.8mmol), 2-bromo-5-methylthiazole (0.18g, 1.02mmol), 1N potassium carbonate solution (5mL), tetrahydrofuran (20mL), Pd(dppf)Cl ₂ (75mg, 0.1mmol) was added In the reaction flask, under the protection of nitrogen, the temperature was raised to 80°C and stirred for 14 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure, 50 mL of ethyl acetate and 50 mL of water were added to the residue for extraction, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane: methanol = 2:1) to obtain 0.30 g of the title product as a black liquid with a yield of 25%.

Step 4: Synthesis of 3-(dimethylphosphono)-5-(5-methylthiazol-2-yl)benzoic acid (6e)

Combine 3-(dimethylphosphono)-5-(5-methylthiazol-2-yl)methyl benzoate (0.25g, 0.81mmol), 1N lithium hydroxide solution (3mL), tetrahydrofuran (5mL), Methanol (5 mL) was sequentially added to the reaction flask and stirred at room temperature for 1 hour. After the reaction, 1N hydrochloric acid solution was added to the reaction solution to adjust the pH to 3-4, and then filtered. The filter cake was washed with 20 mL of water and dried to obtain 80 mg of the title product as a yellow solid with a yield of 34%.

Step 5: (R)-3-(Dimethylphosphono)-5-(5-methylthiazol-2-yl)-N-(1-(2-(trifluoromethyl)pyrimidin-5-yl ) Ethyl) benzamide (compound 6) synthesis

Combine 3-(dimethylphosphono)-5-(-methylthiazol-2-yl)benzoic acid (70mg, 0.24mmol), (R)-1-(2-(trifluoromethyl)pyrimidine-5 -Yl)ethyl-1-amine (60 mg, 0.26 mmol), DIPEA (0.12 g, 0.96 mmol), HATU (0.13 g, 0.34 mmol), and DMF (10 mL) were added to the reaction flask and stirred at room temperature for 1 hour. After the completion of the reaction, 50 mL of ethyl acetate and 50 mL of water were added to the reaction solution for extraction, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (column: Hedea ODS-2 C18, 300mm*250mm, 10μm; eluent: acetonitrile/water=0%-100%) to obtain 39mg of the title product as a white solid with a yield of 35 %.

LC-MS: m/z 469.2 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 1.63 (m, 3H), 1.74 (m, 6H), 2.51 (s, 3H), 5.34 (m, 1H), 7.70 (s, 1H), 8.28 ( m, 1H), 8.36 (m, 1H), 8.47 (s, 1H), 9.14 (s, 2H), 9.34 (m, 1H).

Example 7: (R)-3-(Methyl(piperidin-4-yl)amino)-5-(5-methylthiazol-2-yl)-N-(1-(2-(trifluoromethyl) (Yl)pyrimidin-5-yl)ethyl)benzamide (7)

Step 1: (R)-3-(Methyl(piperidin-4-yl)amino)-5-(5-methylthiazol-2-yl)-N-(1-(2-(trifluoromethyl) )Pyrimidine-5-yl)ethyl)benzamide (Compound 7)

(R)-4-((3-(5-Methylthiazol-2-yl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl) Preparation of (phenyl)aminomethyl tert-butyl)piperidine-1-carboxylic acid tert-butyl ester (7a):

The preparation method is the same as that of Example 1, except that N-tert-butoxycarbonyl-4-piperidone is used instead of oxetan-3-one to prepare Intermediate 7a.

Add (R)-4-((3-(5-methylthiazol-2-yl)-5-((1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl)carbamoyl )Phenyl)aminomethyl tert-butyl)piperidine-1-carboxylic acid tert-butyl ester (400mg, 0.66mmol) and 2M dioxane hydrochloric acid solution (20mL) were added to the reaction flask and stirred at room temperature for 3 hours. After the reaction is over, add saturated sodium bicarbonate aqueous solution to the reaction solution to adjust the pH to 8-9, filter, and pass the filter cake through preparative liquid chromatography (column: Hedea ODS-2 C18, 300mm*250mm, 10μm; elution Reagent: acetonitrile/water=0%-100%) to obtain 135 mg of the title product as a white solid with a yield of 40%.

LC-MS: m/z 505.2 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 1.62 (m, 3H), 1.68 (m, 2H), 1.83 (m, 2H), 2.50 (s, 3H), 2.82 (s, 3H), 2.88 ( m, 2H), 3.20 (m, 2H), 2.97 (m, 1H), 5.28 (m, 1H), 7.33 (m, 2H), 7.61 (m, 2H), 9.12 (s, 2H), 9.17 (s ,1H).

Example 8: (R)-3-(Methyl(1-methylpiperidin-4-yl)amino)-5-(5-methylthiazol-2-yl)-N-(1-(2- (Trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (8) preparation

The preparation method was the same as in Example 1, except that N-methyl-4-piperidone was used instead of oxetan-3-one to obtain the title compound 8.

LC-MS: m/z 519.2 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 1.62 (m, 3H), 1.68 (m, 2H), 1.80 (m, 2H), 2.17 (m, 2H), 2.24 (s, 3H), 2.50 ( s, 3H), 2.82 (s, 3H), 2.94 (m, 2H), 3.70 (m, 1H), 5.28 (m, 1H), 7.28 (m, 1H), 7.38 (m, 1H), 7.60 (m ,2H),9.12(m,3H).

Example 9: (R)-3-(3-methyl-1H-pyrazol-1-yl)-5-(oxetan-3-yloxy)-N-(1-(2- (Trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (9) preparation

Step 1: Synthesis of methyl 3-hydroxy-5-(3-methyl-1H-pyrazol-1-yl)benzoate (9b)

Methyl 3-bromo-5-hydroxybenzoate (5.58g, 24mmol), 3-methyl-1H-pyrazole (2.98g, 36mmol), K ₂ CO ₃ (6.62g, 48mmol), DMSO (60mL) , CuI (2.28g, 12mmol) and L-proline (2.76g, 24mmol) were added to the reaction flask and stirred at 120°C for 16 hours. After the reaction, the pH of the reaction solution was adjusted to 4-5 with 1N hydrochloric acid, 200 mL of water was added, and extraction was performed with 200 mL of ethyl acetate. The organic phase was washed three times with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 2:1) to obtain 0.64 g of the title product as a yellow solid, yield: 10.7%.

Step 2: Synthesis of methyl 3-(3-methyl-1H-pyrazol-1-yl)-5-(oxetan-3-yloxy)benzoate (9c)

Combine 3-hydroxy-5-(3-methyl-1H-pyrazol-1-yl) methyl benzoate (0.64g, 2.7mmol), oxetane-3-yl 4-methylbenzenesulfonic acid Ester (0.94g, 4.1mmol), cesium carbonate (1.34g, 4.1mmol), and DMF (10mL) were added to the reaction flask and stirred at 110°C for 16 hours. After the reaction was completed, 50 mL of ethyl acetate and 50 mL of water were added to the reaction solution for extraction. The organic phase was washed twice with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 0.5 g of the title product as a yellow oil. Yield: 62.9%.

Step 3: Synthesis of 3-(3-methyl-1H-pyrazol-1-yl)-5-(oxetan-3-yloxy)benzoic acid (9d)

Combine 3-(3-methyl-1H-pyrazol-1-yl)-5-(oxetan-3-yloxy)methyl benzoate (0.5g, 1.9mmol), THF (6mL) , Methanol (6mL) and 1N Lithium hydroxide (6mL) were added to the reaction flask and stirred at room temperature for 16 hours. After the reaction, the pH of the reaction solution was adjusted to 3-4 with 1N hydrochloric acid, and 20 mL of water and 20 mL of ethyl acetate were added for extraction. The organic phase was washed once with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 0.4 g of the title product as a yellow oily substance. Yield: 75.1%.

Step 4: (R)-3-(3-methyl-1H-pyrazol-1-yl)-5-(oxetan-3-yloxy)-N-(1-(2-( Synthesis of trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide (compound 9)

Combine 3-(3-methyl-1H-pyrazol-1-yl)-5-(oxetan-3-yloxy)benzoic acid (220mg, 0.80mmol), DMF (10mL), HATU ( 426mg, 1.12mmol), DIPEA (413mg, 3.20mmol), (R)-1-(2-(trifluoromethyl)pyrimidin-5-yl)ethyl-1-amine (201mg, 0.88mmol) added to the reaction flask In, stirring at room temperature for 16 hours. After the completion of the reaction, 20 mL of ethyl acetate and 20 mL of water were added to the reaction solution for extraction, the organic phase was washed twice with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (column: Hedea ODS-2 C18, 300mm*250mm, 10μm; eluent: 0%-100% acetonitrile: aqueous solution) to obtain 99mg of the title product as a white solid. Yield: 27.6%.

LC-MS: m/z 448.16 [M+H] ⁺ .

¹ H NMR (300MHz, DMSO-d ₆ ) δ 1.61 (m, 3H), 2.28 (m, 3H), 4.57 (m, 2H), 4.96 (m, 2H), 5.28 (m, 1H), 5.45 ( m, 1H), 6.36 (m, 1H), 7.14 (m, 1H), 7.35 (m, 1H), 7.90 (s, 1H), 8.46 (s, 1H), 9.10 (m, 3H).

Example 10: (R)-3-(((4,4-Difluoropiperidin-1-yl)sulfonyl)-5-(5-methylthiazol-2-yl)-N-(1-( Preparation of 2-(trifluoromethyl)pyrimidin-5-yl)ethyl)benzamide

The preparation method was the same as in Example 2, except that 4,4-difluoropiperidine was used instead of tetrahydropyrrole to obtain the title compound 10.

LC-MS: m/z 576.35 [M+H] ⁺ .

¹ H NMR (300MHz, CDCl ₃ ) δ 1.64 (d, J = 7.2Hz, 3H), 2.09 (s, 4H), 2.55 (s, 3H), 3.17 (s, 4H), 5.35-5.30 (m, 1H), 7.74 (s, 1H), 8.30-8.27 (m, 2H), 8.62 (s, 1H), 9.15 (s, 2H), 9.47 (d, J=6.6 Hz, 1H).

Example 11: (R)-3-((6-oxa-2-azaspiro[3.4]octyl-2-yl)sulfonyl)-5-(5-methylthiazol-2-yl)- Preparation of N-(1-(2-(Trifluoromethyl)pyrimidin-5-yl)ethylbenzamide

The preparation method was the same as in Example 2, except that 6-oxa-2-azaspiro[3.4]octane was used instead of tetrahydropyrrole to obtain the title compound 11.

LC-MS: m/z 568.41 [M+H] ⁺ .

¹ H NMR (300MHz, CDCl ₃ ) δ 1.65 (d, J = 7.1 Hz, 3H), 1.87-1.84 (m, 2H), 2.55 (s, 3H), 3.49 (s, 2H), 3.58-3.54 ( m,2H),3.81-3.78(m,4H),5.40(s,1H),7.75(s,1H),8.35-8.33(m,2H),8.56(s,1H),9.15(s,2H) , 9.52 (d, J = 6.9 Hz, 1H).

Example 12: (R)-3-(5-Methylthiazol-2-yl)-5-(1-phosphinyl-1-yl)-N-(1-(2-(trifluoromethyl) Preparation of pyrimidin-5-yl)ethyl)benzamide

Step 1: Synthesis of 1-cyclobutylphosphine oxide (12b):

Add magnesium chips (10.0g, 417mmol) to tetrahydrofuran (400mL) under nitrogen at room temperature, stir for half an hour at room temperature, and then add 1,4-dibromobutane (43.0g, 201mmol) dropwise to it to control the reaction The temperature is lower than 30°C, and after the dropwise addition is completed, stir at room temperature for 1.5 hours. Then, diethyl phosphonate (13.9g, 100mmol) was added dropwise to it, and the reaction temperature was controlled to be lower than 30°C. After the dropwise addition, the reaction was carried out at room temperature for 16 hours. After the reaction, it was quenched with 20% potassium carbonate aqueous solution (300 mL), filtered, and the filtrate was concentrated under reduced pressure. The residue was distilled under reduced pressure to obtain 530 mg of the title compound as a colorless oil, yield: 5.10%.

GC-MS: m/z 104.0[M].

The remaining steps are the same as the preparation method of Example 6, except that 1-cyclobutyl phosphine oxide (12b) is used instead of dimethyl phosphine oxide to obtain the title compound 12.

LC-MS: m/z 494.62 [M+H] ⁺ .

¹ H NMR(400MHz, CDCl ₃ )δ1.77(d,3H), 2.18-2.04(m,8H), 2.59(s,3H), 5.40(s,1H), 7.60(s,1H), 8.28- 8.18 (m, 2H), 8.67 (s, 1H), 9.05 (s, 2H).

Biological evaluation of the compounds of the invention

Test Example 1: Evaluation of the compound of the present invention on the inhibitory activity of human P2X3 receptors

Fluorescence Image Plate Reader (FLIPR, Molecular Devices, 0296) was used to monitor changes in intracellular calcium levels to evaluate the inhibitory activity of the compounds of the present invention on human P2X3 receptors.

Experimental process: Resuscitate HEK293-pCMV6-P2X3 cell line (Pharmaron, Clone#34), and inoculate it in complete medium (DMEM, high glucose (31053028, Gibco) + 10% fetal bovine serum (FBS) ( Gibco, 10099141) + 4mM GlutaMAX (Gibco, 35050-061) + 1× penicillin-streptomycin, liquid (100×, Gibco, 15140-122) + 350μg/ml geneticin (invitrogen, 10131-027), Cultivate in a 37°C, 5% CO ₂ incubator. When the cells are cultured to 70% to 90% confluence, trypsin (Thermofisher, 12604021) digests the cells and resuspends them in cell seeding medium (DMEM, high glucose ( 31053028, Gibco) + 2% Fetal Bovine Serum (FBS) (Gibco, 10099141) + 4mM GlutaMAX (Gibco, 35050-061)), inoculate 11000 cells/well/25μL to a 384-well cell culture plate (Corning, 3845) Component A powder (FLIPR Calcium 6 Assay Kit) was incubated in an incubator at 37°C and 5% CO _{2 for 22 hours.} , Molecular Devices, R8191) Dilute to 2× working concentration, equilibrate to room temperature for later use. Place the 384-well cell culture plate at room temperature to equilibrate for 10 minutes, remove the medium, add 25μL experiment buffer and 25μL 2×Component A, room temperature 200g After centrifugation for 3-5 seconds, let it stand for 2 hours at 37° C. Dilute α, β-MeATP (Sigma, M6517) to 2.1 μM with experimental buffer, transfer 50 μL to a 384-well plate, and place it at room temperature for later use. The cell culture plate is allowed to stand at room temperature for 10 minutes, and the working solution of the compound to be tested (the initial concentration of detection is 10000 nM, and the cell seeding medium is used to dilute 3 times so that the final concentration of DMSO is 0.1%) is added to the corresponding 384-well cell culture plate Incubate the experimental wells at room temperature for 30 minutes. Use FLIPR Tetra (Molecular Devices, 0296) to add 10 μL of diluted α, β-MeATP to the corresponding experimental wells to be tested, set the excitation wavelength to 470-495nm and the emission wavelength to 515- Detect the fluorescence value at 575nm and collect the data.

Nonlinear four parameter fit using GraphPad compound of formula IC _50:

X: log value of compound concentration; Y: ratio.

The inhibitory activity of the compounds of the present invention on P2X3 receptors is shown in Table 1 below.

Table 1. The compound of the present invention inhibits the IC ₅₀ value of P2X3 receptor

化合物Compound	P2X3 IC ₅₀(nM) P2X3 IC ₅₀ (nM)
实施例1Example 1	AA
实施例2Example 2	AA
实施例3Example 3	BB
实施例4Example 4	AA
实施例5Example 5	BB
实施例6Example 6	AA
实施例7Example 7	AA
实施例8Example 8	BB

实施例9Example 9	CC
实施例12Example 12	AA

A: IC ₅₀ ≤100nM, B: 100nM＜IC ₅₀ ≤200nM, C: 200nM＜IC ₅₀

Conclusion: As described in Table 1 above, the compounds of the present invention show P2X3 antagonistic activity in vitro.

Test Example 2: Evaluation of the compound of the present invention on the inhibitory activity of human P2X2/3 receptor

The FLIPR method was used to monitor changes in intracellular calcium levels to evaluate the inhibitory activity of the compounds of the present invention on human P2X2/3 receptors.

Experimental process: The HEK293/hP2X2/3 cell line (Bioduro clone#164) was resuscitated, and the cells were seeded in a 384-well plate coated with 5 μL/well of 1X Matrigel (BD Bioscience, 354230). Incubate for 30 minutes in a 37°C, 5% CO _{2 incubator.} Then process the cells, remove the culture medium, wash once with PBS, digest the cells with 0.25% trypsin-EDTA (Invitrogen, 25200056), and adjust the cell density to 7.5× ^{10 5} /mL. Add the diluted cells to a 384-well detection plate (Corning, 3709), 20 μL/well. Place the well plate in a 37°C, 5% CO ₂ incubator and incubate overnight. The cell culture medium in the 384-well detection plate was discarded, and 20 μL/well of freshly prepared Fluo-8 buffer (AAT Bioquest, 21080) was added. Incubate for 1 hour in a 37°C, 5% CO _{2 incubator protected from light.}

Compounds of different concentrations were prepared, and 5 μL/well was added to the 384-well detection plate. Incubate in a 37°C, 5% CO ₂ incubator for 30 minutes. Prepare 6X EC80 αβ-meATP (TOCRIS, 3209), use Flipr to add 5 μL/well to a 384-well detection plate, and collect data.

The inhibitory activity of the compounds of the present invention on P2X2/3 receptors is shown in Table 2 below.

Table 2. The compound of the present invention inhibits the IC ₅₀ value of P2X2/3 receptor

化合物Compound	P2X2/3 IC ₅₀(nM) P2X2/3 IC ₅₀ (nM)
实施例1Example 1	AA
实施例2Example 2	BB
实施例5Example 5	CC
实施例6Example 6	AA
实施例7Example 7	BB
实施例8Example 8	CC

A: IC ₅₀ ＞250nM, B: 100n＜IC ₅₀ ≤250nM, C: IC ₅₀ ≤100nM

Conclusion: As described in Table 2 above, the compound of the present invention has poor inhibitory activity on P2X2/3 heterodimeric receptors.

Test Example 3: Pharmacokinetic properties of the compound of the present invention

Animals: Wistar male rats, 180-220g, 7-8 weeks old, purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd., SPF grade, animal production license number: SCXK (京) 2016-0011.

Experimental process: The oral dosage of the compound of Example 6 and Example 12 was 3 mg/kg, and the blood sampling point was determined to be 8 minutes, 15 minutes, 30 minutes, 1 hour, 3 hours, 5 hours, 8 minutes before oral administration and after administration. Hours, 10 hours, 12 hours and 24 hours. Animals were anesthetized by inhalation, anesthesia parameters: flow rate: 1.0 L/m, oxygen pressure: 0.1 MPa, solubility: 4.5%, anesthesia time: 3 minutes. After the animal was anesthetized, 0.5 mL of blood was collected from the orbit, and the blood collection tube was added at a volume ratio of 10 mg/mL lithium heparin to plasma at a volume ratio of 1:10 for anticoagulation. After mixing uniformly, centrifuge at 3000 rpm/centrifuge for 10 minutes, take the upper layer of plasma, and freeze it in a refrigerator at -20°C for later use.

Take 50μL of animal plasma sample and place it in a 1.5ml EP tube, add 5μL of internal standard working solution, and vortex for 60 seconds to mix thoroughly. After vortexing, add 0.2 mL of acetonitrile, vortex vigorously for 1 minute, and centrifuge at 16000 rpm for 10 minutes. Pipette 0.2 mL of the supernatant, filter it with a 0.22 μm filter membrane, and add it to the sample vial for testing. Separate and determine under mass spectrometry conditions, and record the peak area of each sample to be tested and the internal standard.

Use data processing software to integrate the analyte and the internal standard to obtain the peak area; take the concentration of the analyte (x) as the abscissa, and the peak area ratio (y) of the analyte and the internal standard as the ordinate, Use the weighted least squares method (weight 1/x2) to perform the regression operation, and the linear regression equation obtained is the plasma calibration curve. DAS was used for statistical analysis. Obtain various pharmacokinetic parameters and drug-time curves.

The pharmacokinetic properties of the compound of Example 6 and Example 12 of the present invention are shown in Table 3 below.

Table 3. Pharmacokinetic parameters of the compounds of the invention

实施例Example	AUC _(0-t)(ug/Lh) AUC _(0-t) (ug/Lh)	t _1/2z(h) t _1/2z (h)	T _max(h) T _max (h)	C _max(ug/L) C _max (ug/L)
实施例6Example 6	＞200＞200	＞5＞5	＞5＞5	＞20＞20
实施例12Example 12	＞200＞200	＞1＞1	＞1＞1	＞20＞20

Test Example 4: Efficacy of the compound of the present invention on histamine-citrate guinea pig acute cough model

Animal: Dunkin Hartley guinea pig, male, 300-350g. Purchased from Beijing Jinmuyang Experimental Animal Breeding Co., Ltd., general grade, animal production license number: SCXK (Beijing) 2015-0005.

Experimental process: The animals were grouped into a model control group, a 3 mg/kg administration group of the compound of Example 6 and a 30 mg/kg administration group of the compound of Example 6. 30-60 minutes before stimulating guinea pigs to cough, each group was given corresponding drugs. First, use 2mg/mL histamine solution ultrasonic atomization inhalation to the allergic guinea pig for 1-2 minutes, and quickly take it out after coughing. After 5 minutes, use 2M citric acid solution to inhale the cough for 5 minutes; from citric acid atomization At the beginning, the guinea pig's incubation period of cough and the number of coughs within 5 minutes were observed, and the antitussive effect of the compound of Example 6 on the guinea pig cough model was evaluated based on the guinea pig's incubation period and the number of coughs.

The curative effect of the compound of the present invention on histamine-citrate guinea pig acute cough model is shown in Table 4.

Table 4. Changes in the incubation period of cough in guinea pigs

Note: *P<0.05v.s model group; **P<0.01v.s model group, T-test

Conclusion: The compound of Example 6 can effectively prolong the incubation period of cough in guinea pigs, significantly reduce the number of coughs, and has the value of further development.

Claims

A compound represented by general formula (I) or its mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof,

in:

W 1 , W 2 , and W 3 are each independently selected from CR 6 or N;

A 1 , A 2 , A 3 , A 4 , and A 5 are each independently selected from C, N, O or S;

R 1 is selected from -NR a R b , -NR a S(O) m R b , -NR a S(O) m NR a R b , -NR a S(O)(NR a )R b , -NR a S(O)(NR a )NR b , -NR a C(O)R b , -NR a C(O)NR a R b , -S(O) m R a , -S(O) m NR a R b , -S(O)(NR a )NR a R b , -OR a , -C(O)NR a R b , -P(O)R a R b , -(CR a R b )R b ;

Each R 2 is independently selected from hydrogen, halogen, amino, nitro, cyano, hydroxyl, mercapto, oxo, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl , -C(O)R a , -O(O)CR a , -C(O)OR a , -C(O)NR a R b , -NHC(O)R a , -S(O) m R a, -S (O) m NR a R b, -NHS (O) m R a; wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl groups are optionally further substituted Selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxy, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, hetero One or more groups of aryl are substituted;

R 3 is selected from aryl or heteroaryl; the aryl or heteroaryl is optionally further substituted with one or more groups selected from halogen, alkyl, haloalkyl, alkoxy, and haloalkoxy;

L is selected from -C(R 4 R 5 )-;

R 4 and R 5 are each independently selected from hydrogen, alkyl, alkoxy; wherein the alkyl and alkoxy are optionally further selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxy, One or more group substitutions of ester group, oxo group, alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

R 6 is selected from hydrogen, halogen, hydroxyl, cyano, amino, carboxy, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein The alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group are optionally further selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxyl, One or more group substitutions of ester group, oxo group, alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

R a and R b are each independently selected from hydrogen, halogen, hydroxyl, cyano, amino, carboxy, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, Heteroaryl groups, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups are optionally further selected from halogen, amino, nitro, cyano, One of hydroxyl, mercapto, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl or one of Multiple group substitutions;

Or R a and R b together with the atoms to which they are attached form a cycloalkyl or heterocyclic group, the cycloalkyl or heterocyclic group is optionally further selected from halogen, amino, nitro, cyano, oxo, One or more groups of hydroxyl, mercapto, carboxyl, ester, alkyl, alkoxy, haloalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl replace;

m is 0, 1 or 2;

n is an integer from 0 to 3.
The compound represented by the general formula (I) according to claim 1 or its mesomer, racemate, enantiomer, diastereomer, or mixture form thereof, or pharmacologically thereof Salt for use, which is a compound represented by the general formula (II) or its mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof ,

Wherein, A 1 , A 2 , A 3 , A 4 , A 5 , R 1 , R 2 , R 3 , L, and n are as defined in claim 1.
The compound represented by the general formula (I) according to claim 1 or 2, or its mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or Medicinal salt,

in,

Group
Selected from pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isothiazolyl, isoxazolyl, 1,3,4-oxadiazolyl, 1,2,4 -Oxadiazolyl, 1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl, triazolyl or tetrazolyl, preferably pyrazolyl, thiazolyl, oxazolyl or 1, 3,4-oxadiazolyl, more preferably thiazolyl;

Said
Replaced by n R 2;

R 2 and n are as defined in claim 1.
The compound represented by the general formula (I) according to any one of claims 1 to 3, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or its pharmaceutically acceptable salt,

Wherein, R 3 is a C 6 -C 10 aryl group or a 5- to 10-membered heteroaryl group, preferably phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, more preferably pyrimidinyl, the aryl or hetero an aryl group optionally further substituted selected from halogen, C 1- C 6 alkyl, C 1- C 6 haloalkyl, C 1- C 6 alkoxy, C 1- C 6 haloalkoxy groups, one or more replace.
The compound represented by the general formula (I) according to any one of claims 1 to 4, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from -NR a R b , -NR a S(O) m R b , -NR a S(O) m NR a R b , -NR a S(O)(NR a )R b , -NR a S(O)(NR a )NR b , -NR a C(O)R b , -NR a C(O)NR a R b , -S(O) m R a , -S(O) m NR a R b , -S(O)(NR a )NR a R b , -OR a , -C(O)NR a R b , -P(O)R a R b , -(CR a R b )R b , preferably -NR a R b , -NR a S(O) 2 R b , -NR a S(O)(NR a )R b , -NR a S(O) 2 NR a R b , -NR a S(O)(NR a )NR b , -NR a C(O)R b , -NR a C(O)NR a R b , -S(O)R a , -SO 2 R a , -S( O) 2 NR a R b , -S(O)(NR a )NR a R b , -OR a , -P(O)R a R b ;

R a and R b are each independently selected from hydrogen, halogen, alkyl, alkoxy, cycloalkyl, and heterocyclyl, wherein the alkyl, alkoxy, cycloalkyl, and heterocyclyl are optionally further selected from Selected from halogen, amino, nitro, cyano, hydroxyl, mercapto, carboxy, ester, oxo, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, hetero One or more aryl groups are substituted;

Or R a and R b together with the atoms to which they are attached form a 5- to 8-membered heterocyclic group, the heterocyclic group is optionally further selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, One or more group substitutions of carboxyl group, ester group, alkyl group, alkoxy group, alkenyl group, alkynyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

m is 0, 1, or 2.
The compound represented by the general formula (I) according to any one of claims 1 to 5, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from -NR a R b ;

R a is selected from hydrogen, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 5-8 membered heterocyclyl particularly oxygen Etanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, wherein the C 1 -C 6 alkyl, C 3 -C 6 ring Alkyl and 5-8 membered heterocyclic groups are optionally further selected from halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy One or more groups of the group are substituted;

R b is selected from hydrogen or C 1 -C 6 alkyl.
The compound represented by the general formula (I) according to any one of claims 1 to 5, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from -NR a R b ;

R a and R b together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocyclic group, preferably tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, or 8-membered spiro heterocyclic group; The cyclic group is optionally further selected from one or more groups selected from halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy replace.
The compound represented by the general formula (I) according to any one of claims 1 to 5, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected -S (O) R a, or -SO 2 R a;

R a is selected from hydrogen, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 5-8 membered heterocyclyl particularly oxygen Etanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, wherein the C 1 -C 6 alkyl, C 3 -C 6 ring Alkyl and 5-8 membered heterocyclic groups are optionally further selected from halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy One or more groups of the group are substituted.
The compound represented by the general formula (I) according to any one of claims 1 to 5, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from -S(O) 2 NR a R b ;

R a is selected from hydrogen, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 5-8 membered heterocyclyl particularly oxygen Etanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, wherein the C 1 -C 6 alkyl, C 3 -C 6 ring Alkyl and 5-8 membered heterocyclic groups are optionally further selected from halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy One or more groups of the group are substituted;

R b is selected from hydrogen or C 1 -C 6 alkyl;

Alternatively, R a and R b together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocyclic group, preferably tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, or 8-membered spiro heterocyclic group; The heterocyclic group is optionally further selected from one or more of halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy Group substitution.
The compound represented by the general formula (I) according to any one of claims 1 to 5, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from -OR a ;

R a is selected from hydrogen, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 5-8 membered heterocyclyl particularly oxygen Etanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, wherein the C 1 -C 6 alkyl, C 3 -C 6 ring Alkyl and 5-8 membered heterocyclic groups are optionally further selected from halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy One or more groups of the group are substituted.
The compound represented by the general formula (I) according to any one of claims 1 to 5, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from -P(O)R a R b ;

R a is selected from hydrogen, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 5-8 membered heterocyclyl particularly oxygen Etanyl, azetidinyl, tetrahydrofuranyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, wherein the C 1 -C 6 alkyl, C 3 -C 6 ring Alkyl and 5-8 membered heterocyclic groups are optionally further selected from halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 1 -C 6 haloalkoxy One or more groups of the group are substituted;

R b is selected from hydrogen or C 1 -C 6 alkyl;

Or R a and R b together with the atoms to which they are attached form a 5- to 8-membered heterocyclic group, especially a phosphoryl group; the heterocyclic group is optionally further selected from halogen, C 1 -C 6 alkyl, One or more groups of C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, and C 1 -C 6 haloalkoxy are substituted.
The compound represented by the general formula (I) according to any one of claims 1 to 11, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

R 2 is selected from hydrogen, halogen, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 3 -C 7 cycloalkyl, 5- to 7-membered heterocyclic group; preferably C 1 -C 6 alkyl ; Wherein the alkyl group, alkoxy group, cycloalkyl group, heterocyclic group are optionally further selected from halogen, amino, nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, ester, alkyl, One or more groups of alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are substituted.
The compound represented by the general formula (I) according to any one of claims 1 to 12, or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, wherein

L is selected from -C(R 4 R 5 )-;

R 4 and R 5 are each independently selected from hydrogen, C 1 -C 6 alkyl, and C 1 -C 6 alkoxy.
The compound represented by the general formula (I) according to any one of claims 1 to 13 or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, the compound is selected from:
A method for preparing the compound represented by the general formula (I) according to any one of claims 1 to 14 or its mesomer, racemate, enantiomer, diastereomer, Or its mixture form, or its pharmaceutically acceptable salt method, which comprises the following steps:

In the presence of a condensing agent, the compound Ig and Ih are subjected to a condensation reaction under basic conditions to obtain a compound of general formula (I), wherein the basic condition is preferably DIPEA, and the condensing agent is preferably HATU;

Wherein, W 1 , W 2 , W 3 , A 1 , A 2 , A 3 , A 4 , A 5 , R 1 , R 2 , R 3 , L, n are as defined in claim 1.
A pharmaceutical composition containing the compound represented by the general formula (I) according to any one of claims 1 to 14 or its meso, racemate, enantiomer, or non-pair Enantiomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.
The compound represented by the general formula (I) according to any one of claims 1 to 14 or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, or use of the pharmaceutical composition according to claim 16 in the preparation of a P2X3 antagonist.
The compound represented by the general formula (I) according to any one of claims 1 to 14 or a meso, racemate, enantiomer, diastereomer, or a mixture thereof Form, or a pharmaceutically acceptable salt thereof, or use of the pharmaceutical composition according to claim 16 in the preparation of a medicament for the prevention and/or treatment of diseases related to P2X3 activity.
The use according to claim 18, wherein the disease related to P2X3 activity is selected from the group consisting of respiratory system disease, urogenital system disease, digestive system disease, nervous system disease, musculoskeletal system disease, circulatory system disease and pain; Respiratory diseases such as: chronic obstructive pulmonary disease, emphysema, asthma, bronchospasm, pulmonary fibrosis, acute cough, chronic cough; the genitourinary system diseases such as: endometriosis, uterine fibroids, dysmenorrhea, Pelvic inflammatory diseases, urethritis, cystitis, overactive bladder, urinary incontinence, prostatic hyperplasia, prostatitis, orchitis, dyspareunia; the digestive system diseases such as: irritable bowel syndrome, ulcerative colitis, gram Rohn’s disease, biliary colic and other biliary diseases, functional bowel disease, gastroesophageal reflux, gastric dilation, and colon dilatation; the nervous system diseases such as: epilepsy, neuritis, neuropathy, Alzheimer’s disease, Parkinson’s Diseases; the musculoskeletal system diseases such as: gout, arthritis such as osteoarthritis, rheumatoid arthritis and ankylosing spondylitis; the circulatory system diseases such as: cerebral ischemia, traumatic brain injury, myocardial infarction.