SLC6A19 inhibitor compound, pharmaceutical composition as well as preparation method and application of SLC6A19 inhibitor compound and pharmaceutical composition

Abstract

The invention provides an SLC6A19 inhibitor compound, a pharmaceutical composition as well as a preparation method and application of the SLC6A19 inhibitor compound. The compound has a good SLC6A19 inhibition effect, is used for treating SLC6A19 mediated diseases and / or diseases, such as phenylketonuria, and is used for preparing medicines for the diseases or the diseases. # imgabs0 #

Description

[0001] This invention claims the priority of the prior applications titled "SLC6A19 Inhibitor Compounds, Pharmaceutical Compositions, and Their Preparation Methods and Applications", with patent application number 202311753168.5, filed with the China National Intellectual Property Administration on December 19, 2023; and titled "SLC6A19 Inhibitor Compounds, Pharmaceutical Compositions, and Their Preparation Methods and Applications", with patent application number 202410302063.6, filed with the China National Intellectual Property Administration on March 15, 2024. The entire texts of the above prior applications are incorporated into this invention by reference. Technical Field

[0002] This invention belongs to the field of medicine, and specifically relates to an SLC6A19 inhibitor compound, a pharmaceutical composition, and their preparation methods and applications. Background Art

[0003] Phenylketonuria (PKU) is a rare autosomal recessive genetic disease. Due to gene mutations in phenylalanine hydroxylase (PAH) in children, the activity of PAH decreases or is lost, phenylalanine cannot be converted into tyrosine, the metabolism of phenylalanine is blocked, and a large amount of abnormal products accumulate in the body and cause the disease.

[0004] Newborns with PKU generally do not have special manifestations. Infants who are not treated start to gradually develop the disease 3 - 4 months after birth. Their hair turns from black to yellow, their skin color becomes lighter, and their urine and sweat emit a mousy odor. Adults with PKU may experience lower limb spasms, cerebellar ataxia, tremors, encephalopathy, and vision abnormalities, and severe cases may lead to death.

[0005] The PAH gene is located on chromosome 12 (12q22 - q24.2), and the PAH encoded by it is responsible for metabolizing phenylalanine and maintaining the homeostasis of phenylalanine in the body. Most patients have low or no protein activity due to PAH mutations. In a small number of patients, the folding and assembly of PAH monomers are disrupted due to the deficiency of dihydropteridine reductase (DHPR) or the deletion of the chaperone protein DNAJC12, resulting in an increase in phenylalanine levels.

[0006] Currently, for PKU patients, dietary treatment and drug treatment are mainly carried out. Dietary treatment mainly involves supplementing amino acid mixtures without phenylalanine and consuming low - protein foods. However, long - term consumption of foods lacking protein or phenylalanine can lead to growth restriction, anorexia, hair loss, lethargy, and eczema outbreaks. Sapropterin hydrochloride is mainly used for the treatment of mild PKU patients or BH4 - deficient patients, and the response rate for severe PKU patients is less than 10%; pegvaliase is mainly used for the treatment of moderate - to - severe PKU patients and needs to be injected subcutaneously every day, and long - term injection is likely to produce adverse immune reactions.

[0007] Amino acid transporters mediate the transmembrane transport of amino acids and play an important role in amino acid nutrition of body cells and amino acid metabolism. SLC6A19 is the only transporter for phenylalanine in the apical membranes of small intestinal enterocytes and renal tubules, responsible for absorbing most of the free phenylalanine in the small intestine and reabsorbing phenylalanine by renal proximal tubular cells. Inhibiting SLC6A19 will help inhibit the reabsorption of phenylalanine by renal tubules and reduce the damage to the body caused by phenylalanine metabolic disorders. SUMMARY OF THE INVENTION

[0008] To solve the above technical problems, the present invention provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or its prodrug compound:

[0009]

[0010] Wherein,

[0011] R1 is selected from H, CN, -OH, the following unsubstituted or optionally substituted by one, two or more Rs d substituted groups: C 1-12 alkyl, C 3-12 cycloalkyl, 3- to 14-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl; each R d is the same or different and independently of one another is selected from halogen, C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkyl;

[0012] R2 is selected from the following unsubstituted or optionally substituted by one, two or more Rs b substituted groups: -N(R 21 )(R 22 ), C 1-12 alkyl, C 1-12 alkoxy, C 3-12 cycloalkyl or 3- to 14-membered heterocyclic group; R 21 , R 22 is the same or different and independently of one another is selected from H, C 1-12 alkyl, C 1-12 alkoxy, halo C 1-12 alkyl, halo C 1-12 alkoxy, C 3-12 cycloalkyl, halo C 3-12 cycloalkyl, 3- to 14-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl; or, R 21 , R 22 and the N atom to which it is attached form a 3- to 14-membered heterocyclic ring; each R bIdentical or different, and each independently selected from OH, CN, halogen, oxo (=O), unsubstituted or optionally substituted by one, two or more R b1 substituted with the following groups: C 1-12 alkyl, C 1-12 alkoxy, halo C 1-12 alkyl, C 3-12 cycloalkyl, 3- to 14-membered heterocyclic group, C 6-10 aryl, 5- to 10-membered heteroaryl, -NH2 or -S(O)2-C 1-12 alkyl; each R b1 identical or different, and each independently selected from OH, NH2, CN, halogen, C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkyl;

[0013] R3 and R4 are identical or different, and each independently selected from H, halogen, unsubstituted or optionally substituted by one, two or more substituents selected from halogen, CN, OH, C 1-12 alkyl, halo C 1-12 alkyl-substituted with the following groups: C 1-12 alkyl, C 1-12 alkoxy, halo C 1-12 alkyl, halo C 1-12 alkoxy, C 3-12 cycloalkyl, halo C 3-12 cycloalkyl, 3- to 14-membered heterocyclic group, C 6-10 aryl or 5- to 10-membered heteroaryl;

[0014] L1 is absent or selected from unsubstituted or optionally substituted by one, two or more R e substituted with the following groups: C 1-12 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, C 3-14 cycloalkylene, C 6-14 arylene, 5- to 14-membered heteroarylene, C 6-14 arylene-C 1-12 alkylene or 5- to 14-membered heteroarylene-C 1-12 alkylene; each R e identical or different, and each independently selected from H, OH, CN, halogen, NH2, C 1-12 alkyl, C 1-12 alkoxy, halo C 1-12 alkyl, halo C 1-12 alkoxy, C 3-12 cycloalkyl, halo C 3-12 cycloalkyl;

[0015] L2 is absent or selected from C1-12 Alkylene, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-14 Cycloalkylene, C 6-10 Arylene or 5 - 10 - membered heteroarylene;

[0016] Y1 is selected from the following unsubstituted or optionally substituted by one, two or more R c substituted groups: C 3-14 Cycloalkyl, 3 - 14 - membered heterocyclic group, C 6-10 Aryl or 5 - 10 - membered heteroaryl; each R c is the same or different and independently selected from H, OH, CN, halogen, the following unsubstituted or optionally substituted by one, two or more R c1 substituted groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 1-12 Alkylthio, C 3-12 Cycloalkyl, 3 - 14 - membered heterocyclic group, C 6-10 Aryl, 5 - 10 - membered heteroaryl, -NH2 or -S(O)2 - C 1-12 alkyl; each R c1 is the same or different and independently selected from H, OH, NH2, CN, halogen, C 1-6 alkyl or C 1-6 alkoxy;

[0017] each R a is the same or different and independently selected from H, CN, oxo(=O), halogen, OH, the following unsubstituted or optionally substituted by one, two or more R a1 substituted groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3 - 14 - membered heterocyclic group, C 6-10 Aryl, 5 - 10 - membered heteroaryl, -C(O)N(R a11 )(R a12 ), -N(R a13 )(R a14 ), -S(O)2 - R a15 , -S(O)(=NR a16 )(R a17 ), or -P(O)(R a18 )(R a19 ); or, two Rs attached to the same carbon atom together with the carbon atom to which they are attached form an unsubstituted or optionally substituted by one, two or more R a substituted ring system as follows: C a1 3-14 ​A carbocyclic ring or a 3- to 14-membered heterocyclic ring; or two Rs attached to adjacent carbon atoms a together with the carbon atoms to which they are respectively attached form an unsubstituted or optionally substituted by one, two or more Rs a1 substituted ring system as follows: C 3-14 carbocyclic ring, 3- to 14-membered heterocyclic ring, C 6-14 aryl ring or 5- to 14-membered heteroaryl ring; or two non-adjacent Rs a linked by their terminal groups together form an unsubstituted or optionally substituted by one, two or more Rs a1 substituted C 1-3 alkylene; each R a1 is the same or different and independently of one another is selected from H, OH, CN, halogen, an unsubstituted or optionally substituted by one, two or more Rs a2 substituted group: C 1-12 alkyl, C 1-12 alkoxy, halo C 1-12 alkyl, C 3-12 cycloalkyl, 3- to 14-membered heterocyclic group, C 6-10 aryl, 5- to 10-membered heteroaryl, -NH2 or -S(O)2-C 1-12 alkyl; each R a2 is the same or different and independently of one another is selected from H, OH, NH2, CN, halogen, C 1-6 alkyl, C 1-6 alkoxy or halo C 1-6 alkyl; R a11 , R a12 , R a13 , R a14 , R a15 , R a16 , R a17 , R a18 , R a19 is the same or different and independently of one another is selected from H, C 1-12 alkyl, C 3-8 cycloalkyl or 3- to 8-membered heterocyclic group;

[0018] m is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0019] n is selected from 0, 1, 2 or 3.

[0020] According to some embodiments, each R a is the same or different and independently of one another is selected from H, CN, oxo(=O), halogen, OH, an unsubstituted or optionally substituted by one, two or more Rs a1 substituted group: C 1-12 alkyl, C 1-12 alkoxy, C 3-12 cycloalkyl, 3- to 14-membered heterocyclic group, C6-10 aryl, 5- to 10-membered heteroaryl, -C(O)N(R a11 )(R a12 )、-N(R a13 )(R a14 )、-S(O)2-R a15 、-S(O)(=NR a16 )(R a17 ) or -P(O)(R a18 )(R a19 );each R a1 is the same or different and is independently selected from H, OH, CN, halogen, the following groups which are unsubstituted or optionally substituted by one, two or more R a2 : C 1-12 alkyl, C 1-12 alkoxy, halo-C 1-12 alkyl, C 3-12 cycloalkyl, 3- to 14-membered heterocyclic group, C 6-10 aryl, 5- to 10-membered heteroaryl, -NH2 or -S(O)2-C 1-12 alkyl;each R a2 is the same or different and is independently selected from H, OH, NH2, CN, halogen, C 1-6 alkyl, C 1-6 alkoxy or halo-C 1-6 alkyl;R a11 , R a12 , R a13 , R a14 , R a15 , R a16 , R a17 , R a18 , R a19 is the same or different and is independently selected from H, C 1-12 alkyl, C 3-8 cycloalkyl or 3- to 8-membered heterocyclic group.

[0021] According to some embodiments, m is selected from 0, 1 or 2.

[0022] According to some embodiments, each R a is the same or different and is independently selected from CN, oxo(=O), halogen (e.g., F, Cl, Br), OH, C 1-6 alkyl, C 1-6 alkoxy, halo-C 1-6 alkyl, halo-C 1-6 alkoxy, C 3-6 cycloalkyl or 3- to 6-membered heterocyclic group;alternatively, two R a connected to adjacent carbon atoms together with the carbon atoms to which they are respectively connected form a C 3-6 carbocyclic ring.

[0023] According to some embodiments, R a is OH; alternatively, two Rs attached to adjacent carbon atoms a together with the carbon atoms to which they are respectively attached form a cyclopropyl ring.

[0024] According to some embodiments, R1 is selected from H, CN, -OH, the following groups which are unsubstituted or optionally substituted by one or two Rs d : C 1-6 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocyclic group, phenyl or 5- to 6-membered heteroaryl; each R d is the same or different and is independently selected from C 1-3 alkyl.

[0025] According to some embodiments, R1 is selected from H, CN, -OH, C 1-4 alkyl (such as methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl), C 3-6 cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl), 4- to 6-membered heterocyclic group (such as ), phenyl or 5- to 6-membered heteroaryl (such as ).

[0026] According to some embodiments, R1 is selected from H, CN, -OH, methyl, ethyl, cyclopropyl, cyclopentyl,

[0027] According to some embodiments, R1 is selected from CN or OH.

[0028] According to some embodiments, R2 is selected from the following groups which are unsubstituted or optionally substituted by one, two or more Rs b : -N(R 21 )(R 22 ), C 1-12 alkyl, C 3-12 cycloalkyl or 3- to 14-membered heterocyclic group.

[0029] According to some embodiments, R2 is selected from the following groups which are unsubstituted or optionally substituted by one, two or more Rs b : -N(R 21 )(R 22 ), C 1-6 alkyl (such as methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl), C 3-6 cycloalkyl (such as cyclopropyl) or 4- to 6-membered heterocyclic group (such as ).

[0030] According to some embodiments, R 21 , R22 are the same or different and are each independently selected from H, C 1-12 alkyl, C 3-12 cycloalkyl, a 3- to 14-membered heterocyclic group, C 6-10 aryl, or a 5- to 10-membered heteroaryl; or, R 21 , R 22 and the N atom to which it is attached form a 3- to 14-membered heterocycle.

[0031] According to some embodiments, R 21 , R 22 are the same or different and are each independently selected from H, methyl, ethyl, cyclopropyl,

[0032] R 21 , R 22 and the N atom to which it is attached form

[0033] According to some embodiments, R 21 , R 22 are the same or different and are each independently selected from H, C 1-6 alkyl (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, or tert-butyl), or, R 21 , R 22 and the N atom to which it is attached form

[0034] According to some embodiments, R 21 is selected from H, R 22 is selected from C 1-6 alkyl (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, or tert-butyl).

[0035] According to some embodiments, R2 is selected from the following groups which are unsubstituted or optionally substituted by one, two, or more R b groups: methyl, ethyl, cyclopropyl, -NH2, -NHCH3, -NHCH2CH3,

[0036] According to some embodiments, R2 is selected from the following groups which are unsubstituted or optionally substituted by one, two, or more R b groups: methyl, ethyl, cyclopropyl, -NH2, -NHCH3,

[0037] According to some embodiments, each Rb Identical or different, and each independently selected from OH, CN, halogen or C 1-12 alkyl group.

[0038] According to some embodiments, each R b Identical or different, and each independently selected from OH, CN, F, C 1-6 alkyl group (such as methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl).

[0039] According to some embodiments, L2 is absent.

[0040] According to some embodiments, R3 and R4 are identical or different, and each independently selected from H, halogen, C 1-12 alkyl group, C 1-12 alkoxy group, halo C 1-12 alkyl group, halo C 1-12 alkoxy group, C 3-12 cycloalkyl group, halo C 3-12 cycloalkyl group, 3- to 14-membered heterocyclic group, C 6-10 aryl group or 5- to 10-membered heteroaryl group.

[0041] According to some embodiments, R3 is selected from H.

[0042] According to some embodiments, R4 is selected from unsubstituted or optionally substituted by a substituent selected from halogen (such as F, Cl, Br) or C 1-4 alkyl group (such as methyl, ethyl, isopropyl, tert-butyl) substituted cyclopropyl group.

[0043] According to some embodiments, R4 is selected from

[0044] According to some embodiments, R4 is selected from cyclopropyl group.

[0045] According to some embodiments, L1 is absent or selected from C 1-12 alkylene group, C 2-6 alkenylene group, C 2-6 alkynylene group, C 3-14 cycloalkylene group, C 6-10 arylene group or 5- to 10-membered heteroarylene group.

[0046] According to some embodiments, L1 is selected from optionally substituted by 1 substituent selected from C 1-4 alkyl group (such as methyl, ethyl, isopropyl, tert-butyl) or OH substituted C 1-6 alkylene group or 5- to 6-membered heteroarylene-C 1-6 alkylene group.

[0047] According to some embodiments, L1 is selected from methylene group, -CH(CH3)-, -CH(CH2OH)-, or One side of "*" is connected to Y1, and one side of "#" is connected to N.

[0048] According to some embodiments, L1 is selected from methylene.

[0049] According to some embodiments, Y1 is selected from unsubstituted or optionally substituted by one, two or more R c substituted phenyl, naphthyl or 5-6 membered heteroaryl; each R c is the same or different and independently of one another is selected from H, CN, halogen, unsubstituted or optionally substituted by one, two or more R c1 substituted groups as follows: C 1-6 alkyl, C 1-6 alkoxy or C 1-6 alkylthio; each R c1 is the same or different and independently of one another is selected from H, CN or halogen.

[0050] According to some embodiments, Y1 is selected from unsubstituted or optionally substituted by one, two or more R c substituted phenyl or 5-6 membered heteroaryl; each R c is the same or different and independently of one another is selected from H, CN, halogen, unsubstituted or optionally substituted by one, two or more R c1 substituted groups as follows: C 1-6 alkyl or C 1-6 alkoxy; each R c1 is the same or different and independently of one another is selected from H, CN or halogen.

[0051] According to some embodiments, Y1 is selected from unsubstituted or optionally substituted by one, two or three R c substituted phenyl, naphthyl, pyridyl (such as ), pyrazolyl (such as ), thiazolyl (such as ) or thiophenyl (such as ).

[0052] According to some embodiments, each R c is the same or different and independently of one another is selected from F, Cl, Br, CN, CH3, CF3, CH2CF3, CF2CF3, OCH3, OCF3, OCF2H, OCF2Br, OCF2Cl or SCF3.

[0053] According to some embodiments, Y1 is selected from unsubstituted or optionally substituted by one or two R c substituted phenyl; each R c is the same or different and independently of one another is selected from H, F, Cl, CN, CH3, OCH3 or OCF3.

[0054] According to some embodiments, Y1 is selected from

[0055]

[0056] According to some embodiments, Y1 is selected from According to some embodiments, the compound represented by formula (I) has the structure shown below:

[0057]

[0058]

[0059] wherein, R1, R2, R3, R4, Y1, L1, R c have the definitions as described herein. According to some embodiments, the compound represented by formula (I) has the structure shown below:

[0060]

[0061] wherein, R1, R2 have the definitions as described herein.

[0062] According to some embodiments, the compound represented by formula (I) has the structure shown below:

[0063]

[0064] wherein, Y1, L1, R3, R4, R 21 , R 22 have the definitions as described herein.

[0065] According to some embodiments, the compound represented by formula (I) is selected from the following structures:

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074] The present invention also provides a method for preparing the compound represented by formula (I), comprising the following step A:

[0075]

[0076] Wherein, R1, R2, R3, R4, R a , L1, L2, Y1, m, n have the definitions described herein.

[0077] The present invention also provides a pharmaceutical composition, which comprises at least one of a therapeutically effective amount of the compound represented by formula (I), its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or its prodrug compound.

[0078] According to some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0079] According to some embodiments, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0080] The present invention also provides a method for treating or preventing an SLC6A19-mediated disease or disorder, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or its prodrug compound.

[0081] The present invention also provides a method for treating or preventing an SLC6A19-mediated disease or disorder, comprising administering to a patient a prophylactically or therapeutically effective amount of the above-mentioned pharmaceutical composition.

[0082] According to some embodiments, the SLC6A19-mediated disease or disorder is phenylketonuria.

[0083] According to some embodiments, the patient includes mammals, preferably humans.

[0084] The present invention also provides at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or its prodrug compound, or its pharmaceutical composition for treating or preventing an SLC6A19-mediated disease or disorder.

[0085] According to some embodiments, the SLC6A19-mediated disease or disorder is phenylketonuria.

[0086] The present invention also provides the use of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or its prodrug compound in the preparation of a drug.

[0087] According to some embodiments, the use may be for the preparation of a medicament for treating or preventing an SLC6A19-mediated disease or disorder.

[0088] According to some embodiments, the SLC6A19-mediated disease or disorder is phenylketonuria.

[0089] Beneficial effects

[0090] The compounds provided by the present invention have good SLC6A19 inhibitory effects, can be used for treating or preventing SLC6A19-related disorders and diseases, and for preparing medicaments for treating or preventing such disorders and diseases.

[0091] Term definitions and explanations

[0092] Unless otherwise specified, the group and term definitions described in the specification and claims of the present application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be combined and combined with each other arbitrarily. The group definitions and compound structures after such combinations and combinations should be understood to be within the scope described in the specification and / or claims of the present application.

[0093] The term "optional" (or "optionally", "option") in the general formula definition of the present application means a situation where it is substituted by zero, one or more substituents. For example, "optionally substituted by one, two or more Rs" means that it may not be substituted by R (unsubstituted) or may be optionally substituted by one, two or more Rs.

[0094] "More than" means three or more.

[0095] Unless otherwise specified, the numerical ranges described in the present specification and claims are equivalent to at least recording each specific integer value therein. For example, the numerical range "1-12" is equivalent to recording each integer value in the numerical range "1-12", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

[0096] The term "C 1-12 alkyl" should be understood to mean a straight-chain and branched-chain alkyl having 1 to 12 carbon atoms, and "C 1-8 alkyl" means a straight-chain and branched-chain alkyl having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and "C 1-6"Alkyl" means straight-chain and branched-chain alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. Examples of said alkyl groups are methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. or their isomers.

[0097] The term "C 3-12 cycloalkyl" should be understood to mean a saturated monovalent monocyclic, bicyclic (such as fused rings, bridged rings, spiro rings) hydrocarbon ring or tricyclic alkane having 3 to 12 carbon atoms, preferably "C 3-10 cycloalkyl", more preferably "C 3-8 cycloalkyl". The term "C 3-12 cycloalkyl" should be understood to mean a saturated monovalent monocyclic, bicyclic (such as bridged rings, spiro rings) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The C 3-12 cycloalkyl can be a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as bornanyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonanyl, 2,6-diazaspiro[3,4]octyl, or a tricyclic hydrocarbon group such as adamantyl.

[0098] The term "C 6-14 aryl" should be understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic (such as fused rings, bridged rings, spiro rings) or tricyclic hydrocarbon ring having 6 to 14 carbon atoms, which can be a monoaromatic ring or polyaromatic rings fused together, preferably "C 6-10 aryl". The term "C 6-14 aryl" should be understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C 6-14 aryl"), especially a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or biphenyl, or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C10 "aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C 13 "aryl"), such as fluorenyl, or a ring having 14 carbon atoms ("C 14 "aryl"), such as anthracenyl. When the C 6-20 "aryl" is substituted, it may be mono-substituted or multi-substituted. Also, there is no restriction on the substitution site, for example, it may be ortho-substituted, para-substituted or meta-substituted.

[0099] The term "5- to 14-membered heteroaryl" is to be understood as including monocyclic, bicyclic (such as fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 14 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O, and S, for example, "5- to 10-membered heteroaryl". The term "5- to 14-membered heteroaryl" is to be understood as including monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, especially 5 or 6 or 9 or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O, and S and, additionally, may be benzo-fused in each case. "Heteroaryl" also refers to groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclic rings, where the point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indazolyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 1-, 4-, 5-, 6-, 7-, or 8-phthalazinyl, 2-, 3-, 4-, 5-, or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7-, or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7-, or 8-cinnolinyl, 2-, 4-, 6-, or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-4aH-carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-carbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-carbolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-pyridinyl, 2-, 3-, 4-, 5-, 6-, 8-, 9-, or 10-phenanthroline, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-phenazine, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenothiazine, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenoxazine, 2-, 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, or 10-benzoisoquinolinyl, 2-, 3-, 4-, or thiopheno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-7H-pyrazino[2,[3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]pyranyl, 2-, 3-, 4-, 5-, 7- or 8-5H-pyrido[2,3-d]phthalazinyl, 1-, 3- or 5-1H-pyrazolo[4,3-d]azolyl, 2-, 4- or 5 4H-imidazo[4,5-d]thiazolyl, 3-, 5- or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5- or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8- or 9-furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl, 2-, 4-, 4-, 5-, 6- or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7- or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-4H-pyrrolo[1,2-b][2]benzazapinyl. Representative fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6- or 7-indolyl, 2-, 3-, 4-, 5-, 6- or 7-benzo[b]thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl and 2-, 4-, 5-, 6- or 7-benzothiazolyl. When the 5- to 14-membered heteroaryl group is linked to other groups to form the compounds of the present invention, the carbon atoms on the 5- to 14-membered heteroaryl ring can be linked to other groups, or the heteroatoms on the 5- to 14-membered heteroaryl ring can be linked to other groups. When the 5- to 14-membered heteroaryl group is substituted, it can be mono-substituted or multi-substituted. And there is no limitation on the substitution site, for example, the hydrogen atom linked to a carbon atom on the heteroaryl ring can be substituted, or the hydrogen atom linked to a heteroatom on the heteroaryl ring can be substituted.,

[0100] Unless otherwise defined, the term "3- to 14-membered heterocyclic group" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a monocyclic ring of 4-, 5-, 6- or 7-membered, a bicyclic ring of 7-, 8-, 9-, 10-, 11- or 12-membered (such as fused rings, bridged rings, spiro rings) or a tricyclic ring system of 10-, 11-, 12-, 13- or 14-membered, and contains at least one, for example, 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S can also be optionally oxidized to various oxidation states to form the states of nitrogen oxides, -S(O)- or -S(O)2-. For example, the "3- to 14-membered heterocyclic group" can be a 3- to 14-membered N-containing heterocyclic group (containing at least one N). Preferably, the heterocyclic group can be selected from "3- to 10-membered heterocyclic groups". The term "3- to 10-membered heterocyclic group" means a saturated or unsaturated non-aromatic ring or ring system and contains at least one heteroatom selected from O, S and N. The heterocyclic group can be linked to the rest of the molecule through any one of the carbon atoms or a nitrogen atom (if present). The heterocyclic group can include fused or bridged rings as well as spiro rings. In particular, the heterocyclic group can include but is not limited to: a 4-membered ring, such as azetidinyl, oxetanyl; a 5-membered ring, such as tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a 7-membered ring, such as diazepanyl. Optionally, the heterocyclic group can be benzo-fused. The heterocyclic group can be bicyclic, for example but not limited to a 5,5-membered ring, such as hexahydrocyclopenta[c]pyrrol-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The heterocyclic group can be partially unsaturated, that is, it can contain one or more double bonds, for example but not limited to dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl or 4H-[1,4]thiazinyl, or, it can be benzo-fused, for example but not limited to dihydroisoquinolinyl. When the 3- to 14-membered heterocyclic group is linked to other groups to form the compounds of the present invention, it can be the carbon atom on the 3- to 14-membered heterocyclic group that is linked to other groups, or it can be the heteroatom on the 3- to 14-membered heterocyclic ring that is linked to other groups. For example, when the 3- to 14-membered heterocyclic group is selected from piperazinyl, it can be the nitrogen atom on piperazinyl that is linked to other groups. Or when the 3- to 14-membered heterocyclic group is selected from piperidinyl, it can be the nitrogen atom on the piperidine ring and the carbon atom at its para-position that are linked to other groups.

[0101] The term "spiro ring" refers to a ring system in which two rings share one ring-forming atom.

[0102] The term "fused ring" refers to a ring system in which two rings share two ring-forming atoms.

[0103] The term "bridged ring" refers to a ring system in which two rings share more than three ring-forming atoms.

[0104] The term "halogen" means fluorine, chlorine, bromine and iodine.

[0105] "Halogenated" means substituted by one or more halogens.

[0106] The term "oxo" means an oxo group (=O) formed by the oxidation of a carbon atom, nitrogen atom or sulfur atom in a substituent.

[0107] The term "alkylamino" means -NH-(alkyl) or -N-(alkyl)2, where alkyl is defined as above. Non-limiting examples of alkylamino include: methylamino, ethylamino, propylamino, isopropylamino, butylamino, dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, etc.

[0108] The term "alkyloxy" means -O-(alkyl), where alkyl is defined as above. Non-limiting examples of alkyloxy include: methoxy, ethoxy, propoxy, butoxy. The alkyloxy can 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, alkyloxy, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy or heterocycloalkyloxy.

[0109] The terms "alkyleneoxy" and "oxyalkylene" mean -alkylene-O- or -O-alkylene-, where alkylene represents a straight-chain or branched-chain saturated divalent hydrocarbon group. The definition of the number of carbon atoms of "alkylene" applies to the definition of "alkyl" above. Those skilled in the art can understand that alkyleneoxy or oxyalkylene can be connected to the rest of the molecule containing it in any direction, that is, the two can be used interchangeably.

[0110] The term "nitroxide" refers to a compound formed by the oxidation of a nitrogen atom in the structure of a tertiary amine or a nitrogen-containing (aromatic) heterocyclic compound.

[0111] Unless otherwise specified, a heterocyclic group, heteroaryl group or heteroarylene group includes all its possible isomeric forms, such as its positional isomers. Thus, for some illustrative and non-limiting examples, forms substituted or bonded to other groups at 1, 2 or more positions among its 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12- positions etc. (if any) may be included, including pyridin-2-yl, pyridin-2-ylidene, pyridin-3-yl, pyridin-3-ylidene, pyridin-4-yl and pyridin-4-ylidene; thienyl or thienylene includes thien-2-yl, thien-2-ylidene, thien-3-yl and thien-3-ylidene; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl.

[0112] A wavy line intersecting a chemical bond Is used to indicate the connection position of a group to other atoms in a molecular structure. As Indicates connection to the 3-position of a pyridyl group. When the connection position of a group is not fixed, taking the pyridyl group as an example, it can be represented by The way is shown, which means it can be connected to any connectable position on the pyridyl group. Unless otherwise specified, similar expressions in this application shall be interpreted in the same way as above.

[0113] In the chemical structure of the compounds described in the present invention, the bond Indicates an unspecified configuration, Indicates an absolute configuration, that is, if there are stereoisomers in the chemical structure, the bond Can be Or simultaneously include Both configurations.

[0114] In the present invention, the compounds involved also include isotopically labeled compounds, which are the same as those shown in Formula I, but in which one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F and Cl, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 32 P, 35 S, 18 F and 36Compounds of the invention containing the above isotopes and / or other isotopes of other atoms, prodrugs thereof, or pharmaceutically acceptable salts of said compounds or said prodrugs are within the scope of the invention. Certain isotopically labeled compounds of the invention, for example compounds incorporating radioactive isotopes such as 3 H and 14 C) can be used for drug and / or substrate tissue distribution assays. Tritium (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred because of their ease of preparation and detectability. Furthermore, substitution with heavier isotopes such as deuterium (i.e., 2 H or D) can provide certain therapeutic advantages (e.g., increased in vivo half-life or reduced dose requirements) resulting from higher metabolic stability and can thus be preferred in certain instances. The presence of hydrogen in the substituents of the invention without separately listing the terms deuterium or tritium does not exclude deuterium or tritium, but can also include deuterium or tritium.

[0115] Those skilled in the art will appreciate that the compounds represented by formula (I) can exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they can form acid addition salts; if these compounds have an acidic center, they can form base addition salts; if these compounds contain both an acidic center (e.g., carboxyl) and a basic center (e.g., amino), they can also form inner salts.

[0116] The compounds of the invention can exist in the form of solvates (such as hydrates), wherein the compounds of the invention contain a polar solvent, particularly for example water, methanol or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be present in stoichiometric or non-stoichiometric ratios.

[0117] Depending on their molecular structure, the compounds of the present invention can be chiral and thus can exist in various enantiomeric forms. Consequently, these compounds can exist in racemic form or in optically active form. The compounds of the present invention cover isomers or mixtures thereof, racemates, in which each chiral carbon has the R or S configuration. The compounds of the present invention or their intermediates can be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art, or can be used in synthesis in this form. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, in the R and S forms, suitable N-protected amino acids (such as N-benzoylproline or N-phenylsulfonylproline), or various optically active camphorsulfonic acids. Chromatographic enantiomeric resolution can also be advantageously carried out with the aid of an optically active resolving agent (such as dinitrobenzoylphenylglycine immobilized on silica gel, cellulose triacetate or other carbohydrate derivatives, or chiral derivatized methacrylate polymers). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, such as hexane / isopropanol / acetonitrile.

[0118] The corresponding stable isomers can be separated according to known methods, such as by extraction, filtration or column chromatography.

[0119] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

[0120] The term "therapeutically effective amount" refers to the amount of an active compound or drug that a researcher, veterinarian, physician or other clinician is seeking to elicit a biological or medical response in a tissue, system, animal, individual or human, and it includes one or more of the following: (1) preventing a disease: for example, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or manifested the pathology or symptoms of the disease. (2) inhibiting a disease: for example, inhibiting a disease, disorder or condition (i.e., preventing the further development of the pathology and / or symptoms) in an individual who is experiencing or manifesting the pathology or symptoms of the disease, disorder or condition. (3) alleviating a disease: for example, alleviating a disease, disorder or condition (i.e., reversing the pathology and / or symptoms) in an individual who is experiencing or manifesting the pathology or symptoms of the disease, disorder or condition. Detailed Description of the Invention

[0121] The technical solutions of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only for illustrative explanation of the present invention and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0122] Unless otherwise specified, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

[0123] Example 1 (R)-N'-Cyano-3-(1-cyclopropyl-3-(4-(trifluoromethoxy)-2-fluorobenzyl)ureido)piperidine-1-carboximidamide (Compound 1)

[0124]

[0125] First step: Synthesis of tert-butyl N-[(2-fluoro-4-(trifluoromethoxy)phenyl)methyl]carbamate (Compound 1-3):

[0126] To a mixed solution of 1-bromo-2-fluoro-4-(trifluoromethoxy)benzene (Compound 1-1, 8 g, 30.89 mmol) in 1,4-dioxane (120 mL) and water (15 mL), add potassium trifluoro[({[(2-methyl-2-propionyloxy)carbonyl]amino}methyl)borate] (Compound 1-2, 8.06 g, 33.98 mmol), palladium(II) acetate (0.35 g, 1.54 mmol), 2-(dicyclohexylphosphino)-2,4,6-triisopropylbiphenyl (1.47 g, 3.09 mmol), and potassium carbonate (12.81 g, 92.67 mmol). Replace the gas, stir the reaction solution at room temperature for 20 minutes under a nitrogen atmosphere, and then stir at 105 °C overnight. After cooling to room temperature, filter, add water (100 mL) to dilute the filtrate, extract with ethyl acetate (150 mL × 3), combine the organic phases, wash with saturated sodium chloride aqueous solution (150 mL × 3), dry over anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain Compound 1-3 (6.6 g).

[0127] MS: (ESI, m / z): 295.1 [M - 56 + ACN + H] + , RT (min): 1.994

[0128] Second step: Synthesis of hydrochloride of (2-fluoro-4-(trifluoromethoxy)phenyl)methanamine (Compound 1-4):

[0129] To a solution of tert-butyl N-[(2-fluoro-4-(trifluoromethoxy)phenyl)methyl]carbamate (Compound 1-3, 6.5 g, 21.02 mmol) in ethyl acetate (35 mL), add hydrogen chloride - ethyl acetate solution (35 mL, 4 M) dropwise, and stir at room temperature for 1 hour. Concentrate the reaction solution to obtain Compound 1-4 (5.1 g).

[0130] MS: (ESI, m / z): 193.0 [M - 16] + , RT(min): 1.191

[0131] Synthesis of N-[(2-fluoro-4-(trifluoromethoxy)phenyl)methyl]-1H-imidazole-1-carboxamide (Compound 1-5):

[0132] To a solution of hydrochloride salt of (2-fluoro-4-(trifluoromethoxy)phenyl)methanamine (Compound 1-4, 1.17 g, 4.78 mmol) in N,N-dimethylformamide (15 mL), 1-(1H-imidazole-1-carbonyl) (0.78 g, 4.78 mmol) and N,N-diisopropylethylamine (1.54 g, 11.95 mmol) were slowly added. The reaction mixture was stirred at room temperature for 16 h. Water (30 mL) was added to dilute the reaction mixture, and it was extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated aqueous sodium chloride solution (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain Compound 1-5 (1.25 g).

[0133] MS: (ESI, m / z): 304.0 [M + H] + , RT(min): 1.416

[0134] Synthesis of (3R)-3-(cyclopropylamino)piperidine-1-carboxylic acid tert-butyl ester (Compound 1-8):

[0135] To a solution of Compound (3R)-3-aminopiperidine-1-carboxylic acid tert-butyl ester (Compound 1-6, 10 g, 49.93 mmol) in 1,4-dioxane (100 mL), (1-ethoxycyclopropyl)oxymethyltrimethylsilane (Compound 1-7, 4.35 g, 24.96 mmol), sodium cyanoborohydride (6.28 g, 99.86 mmol), and acetic acid (6.0 g, 99.86 mmol) were added. The reaction mixture was stirred at 60 °C overnight. After cooling to room temperature, water (150 mL) was added to dilute the reaction mixture, and the pH was adjusted to 9 - 10 with 50% potassium carbonate solution. It was extracted with ethyl acetate (150 mL × 3). The combined organic phases were washed with saturated brine (150 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by reverse-phase chromatography (acetonitrile / water containing 0.05% formic acid) to obtain Compound 1-8 (3.7 g).

[0136] MS: (ESI, m / z): 241.1 [M + H] + , RT(min): 1.210

[0137] Step 5: Synthesis of tert-butyl (R)-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)piperidine-1-carboxylate (Compound 1-9):

[0138] To a solution of N-[(2-fluoro-4-(trifluoromethoxy)phenyl)methyl]-1H-imidazole-1-carboxamide (Compound 1-5, 1.0 g, 3.30 mmol) in acetonitrile (10 mL) was added (3R)-3-(cyclopropylamino)piperidine-1-carboxylate tert-butyl ester (Compound 1-8, 0.79 g, 3.3 mmol) and triethylamine (1.00 g, 9.9 mmol). The mixture was stirred at room temperature for 2 hours. Water (10 mL) was added for dilution, and the mixture was extracted with dichloromethane (20 mL × 3). The combined organic phases were washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain Compound 1-9 (1.5 g).

[0139] MS: (ESI, m / z): 476.1 [M+H] + , RT(min): 2.018

[0140] Step 6: Synthesis of hydrochloride of (R)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-(piperidin-3-yl)urea (Compound 1-10):

[0141] A 4 M solution of hydrogen chloride in 1,4-dioxane (10.0 mL) was added to a solution of (R)-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)piperidine-1-carboxylate tert-butyl ester (Compound 1-9, 1.5 g, 3.15 mmol) in 1,4-dioxane (5 mL). The mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated in vacuo to obtain the crude hydrochloride of Compound 1-10 (1.25 g), which was directly used in the next step.

[0142] MS: (ESI, m / z): 376.1 [M+H] + , RT(min): 0.829

[0143] Step 7: Synthesis of (R)-N'-cyano-3-(1-cyclopropyl-3-(4-(trifluoromethoxy)-2-fluorobenzyl)ureido)piperidine-1-carboximidamide (Compound 1):

[0144] The hydrochloride salt of (R)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-(piperidin-3-yl)urea (Compound 1-10) (50 mg, 0.11 mmol) was dissolved in n-butanol (2 mL), and then sodium dicyanamide (19.59 mg, 0.22 mmol) was added. The mixture was stirred at 60 °C for 16 hours. The reaction solution was concentrated to remove the solvent, and the resulting residue was purified by high-performance liquid chromatography under the following conditions (column specification: Waters 2767 / QDA Column: Sunfire C18 19*250 mm*10 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: from 49% B to 49% B within 16 minutes; detection wavelength: 254 nm / 220 nm; retention time (min): 8.5 - 10.0, to obtain Compound 1 (15.55 mg).

[0145] MS: (ESI, m / z): 443.2 [M+H] + , RT(min): 0.901.

[0146] 1 1H NMR (400 MHz, DMSO-d6) δ 7.42 (t, 1H), 7.33 (d, 1H), 7.23 (d, 1H), 7.05 (s, 2H), 6.95 (t, 1H), 4.31 (d, 2H), 4.09–3.94 (m, 2H), 3.50–3.39 (m, 1H), 3.14 (t, 1H), 2.68–2.60 (m, 1H), 2.48–2.43 (m, 1H), 2.09–1.93 (m, 1H), 1.71 (dd, 2H), 1.45–1.25 (m, 1H), 0.92–0.84 (m, 2H), 0.70–0.62 (m, 2H).

[0147] Example 2 (R)-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N'-hydroxy-piperidine-1-carboximidamide (Compound 2)

[0148]

[0149] First step: Synthesis of (R)-1-(1-cyanopiperidin-3-yl)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)urea (Compound 2-2):

[0150] To a solution of hydrochloride salt (100 mg, 0.27 mmol) of (R)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-(piperidin-3-yl)urea (Compound 1-10) in tetrahydrofuran (3 mL), triethylamine (140 mg, 0.81 mmol) and cyanogen bromide (85.8 mg, 0.81 mmol) were slowly added. The reaction mixture was stirred at room temperature for 5 h. Water (10 mL) was added to dilute the reaction mixture, and it was extracted with ethyl acetate (15 mL × 3). The organic phases were combined, washed with water (15 mL) and saturated sodium chloride aqueous solution (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to obtain Compound 2-2 (75 mg).

[0151] MS: (ESI, m / z): 401.4 [M+H] + , RT(min): 1.801

[0152] Synthesis of the second step (R)-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N'-hydroxy-1-piperidinecarboximidamide (Compound 2):

[0153] To a solution of (R)-1-(1-cyanopiperidin-3-yl)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)urea (Compound 2-2, 70 mg, 0.17 mmol) in dimethyl sulfoxide (3 mL), an aqueous solution of hydroxylamine (0.31 mL, 5.10 mmol, mass ratio: 50%) was slowly added dropwise. The reaction mixture was slowly warmed to 30 °C and stirred for 16 h. Saturated sodium bicarbonate aqueous solution (10 mL) was added to dilute the reaction mixture, and it was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by high performance liquid chromatography under the following conditions (column specification: Sunfire C18, 19*250 mm, 10 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: from 27% B to 27% B in 16 minutes; detection wavelength: 254 nm / 220 nm; retention time (min): 6.00 - 10.00) to obtain Compound 2 (15.76 mg).

[0154] MS: (ESI, m / z): 434.2 [M+H] + , RT(min): 7.859

[0155] 11H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1H), 7.41 (t, 1H), 7.33 (dd, 1H), 7.23 (d, 1H), 6.94 (t, 1H), 5.61 (s, 2H), 4.31 (d, 2H), 3.67–3.45 (m, 3H), 2.88 (t, 1H), 2.47–2.38 (m, 2H), 1.97–1.84 (m, 1H), 1.80–1.69 (m, 1H), 1.67–1.56 (m, 1H), 1.53–1.38 (m, 1H), 0.93–0.80 (m, 2H), 0.72–0.57 (m, 2H).

[0156] Example 3 (R,E)-N'-Cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-methylpiperidine-1-carboximidamide (Compound 3)

[0157]

[0158] First step Synthesis of (R,E)-N'-Cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-methylpiperidine-1-carboximidamide (Compound 3):

[0159] Dissolve (E)-N'-cyano-N-methylcarbamimidothioate (Compound 3-1, 41 mg, 0.32 mmol) in N,N'-dimethylformamide (3 mL), add hydrochloride of (R)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-(piperidin-3-yl)urea (Compound 1-10) (80 mg, 0.21 mmol), triethylamine (85 mg, 0.84 mmol) and silver trifluoromethanesulfonate (70 mg, 0.27 mmol). Stir the reaction solution at 25 °C for 1 hour. Dilute with dichloromethane (60 mL), wash with saturated aqueous sodium bicarbonate (30 mL), wash with brine (30 mL × 2). Dry the organic phase with anhydrous sodium sulfate, filter and concentrate. Purify the crude product by reverse-phase preparative chromatography (conditions are as follows: column specification: Prep-HPLC (Waters2767 / QDA) Column: Sunfire C18 19*250 nm*10 μm; flow rate: 20 mL / min; mobile phase A: 0.1% formic acid / water, mobile phase B:

[0160] acetonitrile; elution gradient: 46 - 50%; retention time: 9.0 - 10.2 min) to obtain Compound 3 (20 mg).

[0161] MS: (ESI, m / z): 457.1 [M+H] +,RT(min):1.729

[0162] 1 1H NMR (400 MHz, DMSO-d6) δ 7.42 (t, 1H), 7.33 (dd, 1H), 7.25–7.21 (m, 1H), 7.21–7.15 (m, 1H), 6.98 (t, 1H), 4.31 (d, 2H), 3.93 (d, 1H), 3.83 (d, 1H), 3.56–3.47 (m, 1H), 3.19 (t, 1H), 2.81 (d, 3H), 2.77–2.68 (m, 1H), 2.48–2.43 (m, 1H), 2.06–1.91 (m, 1H), 1.85–1.76 (m, 1H), 1.75–1.66 (m, 1H), 1.54–1.38 (m, 1H), 0.94–0.85 (m, 2H), 0.71–0.63 (m, 2H).

[0163] Example 4 (R,E)-3-(3-(4-Chloro-2-fluoro-5-methylbenzyl)-1-cyclopropylureido)-N'-cyano-N-methylpiperidine-1-carboxamidine (Compound 4)

[0164]

[0165] First step: Synthesis of N-(4-chloro-2-fluoro-5-methylbenzyl)-1H-imidazole-1-carboxamide (Compound 4-2):

[0166] Dissolve (4-chloro-2-fluoro-5-methylphenyl)methanamine (Compound 4-1, 200 mg, 1.15 mmol) in acetonitrile (5 mL), add N,N'-carbonyldiimidazole (186.47 mg, 1.15 mmol) and triethylamine (349.11 mg, 3.45 mmol). After the addition, stir at room temperature for 1 hour. Monitor the reaction by TLC until completion. Quench the reaction mixture with water (30 mL), extract with ethyl acetate (30 mL × 3), and the resulting crude Compound 4-2 (200 mg) is directly used for the next step.

[0167] Second step: Synthesis of tert-butyl (R)-3-(3-(4-chloro-2-fluoro-5-methylbenzyl)-1-cyclopropylureido)piperidine-1-carboxylate (Compound 4-3):

[0168] Dissolve compound N-(4-chloro-2-fluoro-5-methylbenzyl)-1H-imidazole-1-carboxamide (compound 4-1, 200 mg, 0.75 mmol) in acetonitrile (5 mL), add (3R)-3-(cyclopropylamino)piperidine-1-carboxylic acid tert-butyl ester (compound 1-8, 180.25 mg, 0.75 mmol) and triethylamine (227.68 mg, 2.25 mmol). After addition, stir at 50 °C for 1 hour. Quench the reaction mixture with water (30 mL), extract with ethyl acetate (30 mL×3). Dry the organic phase over anhydrous sodium sulfate and concentrate. Purify the residue by column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 1 / 1) to obtain compound 4-3 (300 mg).

[0169] MS:(ESI,m / z):440.2[M+H] + ,RT(min):2.098.

[0170] Synthesis of the hydrochloride salt of step three (R)-3-(4-chloro-2-fluoro-5-methylbenzyl)-1-cyclopropyl-1-(piperidin-3-yl)urea (compound 4-4):

[0171] Dissolve compound (R)-3-(3-(4-chloro-2-fluoro-5-methylbenzyl)-1-cyclopropylureido)piperidine-1-carboxylic acid tert-butyl ester (compound 4-3, 290 mg, 0.66 mmol) in ethyl acetate (2 mL). Add hydrochloric acid ethyl acetate solution (4 mL, 4N). After addition, stir at room temperature for 1 hour. Concentrate the reaction mixture to remove the solvent to obtain the hydrochloride salt of compound 4-4 (210 mg).

[0172] MS:(ESI,m / z):340.1[M+H] + ,RT(min):0.838.

[0173] Synthesis of step four (R,E)-3-(3-(4-chloro-2-fluoro-5-methylbenzyl)-1-cyclopropylureido)-N'-cyano-N-methylpiperidine-1-carboxamidine (compound 4):

[0174] The hydrochloride salt (70 mg, 0.19 mmol) of compound (R)-3-(4-chloro-2-fluoro-5-methylbenzyl)-1-cyclopropyl-1-(piperidin-3-yl)urea (Compound 4-4) was dissolved in acetonitrile (1.5 mL). (E)-N'-Cyano-N-methylcarbamimidothioate (Compound 3-1, 48 mg, 0.372 mmol), silver trifluoromethanesulfonate (96 mg, 0.37 mmol) and triethylamine (94 mg, 0.93 mmol) were added. After the addition, the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to remove the solvent, and the resulting residue was purified by high performance liquid chromatography under the following conditions (column specifications: Waters 2767 / QDA Column: Sunfire C18 19*250 mm*10 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: from 52% B to 52% B within 16 min; detection wavelength: 254 nm / 220 nm; retention time (min): 8.0–8.8), to obtain Compound 4 (2.02 mg).

[0175] MS: (ESI, m / z): 421.1[M+H] + , RT(min): 1.748.

[0176] 1 1H NMR (400 MHz, CDCl3) δ 7.19 (d, 1H), 7.10 (d, 1H), 6.33 (brs, 1H), 5.73 (t, 1H), 4.41 (d, 2H), 4.39–4.33 (m, 1H), 3.79–3.66 (m, 2H), 3.00 (d, 3H), 2.98–2.91 (m, 1H), 2.89–2.78 (m, 1H), 2.40–2.35 (m, 1H), 2.33 (s, 3H), 2.09–2.01 (m, 2H), 1.86–1.80 (m, 1H), 1.02–0.83 (m, 3H), 0.72–0.68 (m, 2H).

[0177] Example 5 (R,E)-1-(1-((Cyanoimino)(cyclopropyl)methyl)piperidin-3-yl)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)urea (Compound 17)

[0178]

[0179] First step: Synthesis of (E)-methyl N-cyanocyclopropanecarboximidate (Compound 17-2):

[0180] To a solution of methyl cyclopropanecarboximidate (Compound 17-1, 1 g, 10.1 mmol) in methanol (15 mL) was added aminocyanide (424 mg, 10.1 mmol). The reaction mixture was stirred at 25 °C for 3 h, and then aminocyanide (173 mg, 4.1 mmol) was added. The reaction mixture was further stirred at room temperature for 1 h. The insoluble solid was filtered off, and the filtrate was diluted with dichloromethane (100 mL). The combined organic phases were washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to give Compound 17-2 (100 mg).

[0181] MS:(ESI,m / z):125.2[M+H] + ,RT(min):1.437

[0182] Synthesis of the second step (R,E)-1-(1-((cyanoimino)(cyclopropyl)methyl)piperidin-3-yl)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)urea (Compound 17):

[0183] Compound (E)-methyl N-cyanocyclopropanecarboximidate (Compound 17-2, 60 mg, 0.16 mmol) was dissolved in ethanol (3 mL), and hydrochloride salt of compound (R)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-(piperidin-3-yl)urea (Compound 1-10) (30 mg, 0.24 mmol) and N,N'-diisopropylethylamine (62 mg, 0.48 mmol) were added. The reaction mixture was stirred at 70 °C for 2 h. Water (30 mL) was added to quench the reaction, and the mixture was diluted with ethyl acetate (30 mL×3) and washed with brine (30 mL×2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse-phase preparative chromatography (conditions as follows: column specification: Prep-HPLC (Waters 2767 / QDA) Column: Sunfire C18 19*250 nm*10 μm; flow rate: 20 mL / min; mobile phase A: 0.1% formic acid / water, mobile phase B: acetonitrile; elution gradient: 45 - 50%; retention time: 7.0 - 9.2 min) to give Compound 17 (3 mg).

[0184] MS:(ESI,m / z):468.0[M+H] + ,RT(min):1.828

[0185] 11H NMR (400 MHz, DMSO-d6) δ 7.42 (t, 1H), 7.33 (d, 1H), 7.23 (d, 1H), 6.98 (brs, 1H), 4.49–4.14 (m, 4H), 3.66–3.37 (m, 2H), 3.29–3.06 (m, 1H), 3.06–2.77 (m, 1H), 2.49–2.43 (m, 1H), 2.16–2.02 (m, 1H), 1.96–1.70 (m, 3H), 1.08–0.85 (m, 5H), 0.73–0.62 (m, 3H).

[0186] Example 6 (R,E)-N'-Cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-ethylpiperidine-1-carboximidamide (Compound 33)

[0187]

[0188] First step: Synthesis of (E)-N'-cyano-N-ethylcarbamimidothioate (Compound 33-3):

[0189] Dissolve cyanodithioimidocarbonate (Compound 33-1, 500 mg, 3.42 mmol) in ethanol (5 mL), add ethylamine (Compound 33-2, 154.17 mg, 3.42 mmol), and stir the reaction mixture at 110 °C for 1 hour. After cooling, filter and dry the solid to obtain Compound 33-3 (200 mg).

[0190] MS: (ESI, m / z): 143.9 [M+H] + , RT(min): 1.299

[0191] Second step: Synthesis of (R,E)-N'-cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-ethylpiperidine-1-carboximidamide (Compound 33):

[0192] The hydrochloride salt of compound (R)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-(piperidin-3-yl)urea (Compound 1-10) (50 mg, 0.13 mmol) was dissolved in N,N-dimethylformamide (1 mL), and (E)-N'-cyano-N-ethylcarbamimidoyl thioformate (Compound 33-3, 24.20 mg, 0.17 mmol), triethylamine (52.62 mg, 0.52 mmol), and silver trifluoromethanesulfonate (43.42 mg, 0.17 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with saturated aqueous sodium bicarbonate (20 mL) and extracted with ethyl acetate (20 mL × 3). The organic phase was washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by high performance liquid chromatography under the following conditions (column specifications: Waters 2767 / QDA Column: Sunfire C18 19*250 mm*10 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: from 54% B to 54% B in 16 min; detection wavelength: 254 nm / 214 nm; retention time (min): 8.4 - 9.4), to obtain Compound 33 (22 mg).

[0193] MS: (ESI, m / z): 471.1 [M+H] + , RT(min): 1.818

[0194] 1 1H NMR (400 MHz, DMSO-d6) δ 7.42 (t, 1H), 7.33 (d, 1H), 7.23 (d, 1H), 7.19 (t, 1H), 6.99 (t, 1H), 4.31 (d, 2H), 3.94 (d, 1H), 3.81 (d, 1H), 3.56–3.47 (m, 1H), 3.31–3.25 (m, 2H), 3.18 (t, 1H), 2.78–2.69 (m, 1H), 2.47–2.43 (m, 1H), 2.07–1.96 (m, 1H), 1.85–1.67 (m, 2H), 1.52–1.41 (m, 1H), 1.08 (t, J = 7.1 Hz, 3H), 0.93–0.85 (m, 2H), 0.71–0.64 (m, 2H).

[0195] Example 7 (R,E)-N'-Cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-ethylpiperidine-1-carboximidamide (Compound 34)

[0196]

[0197] Step 1: Synthesis of (E)-N'-cyano-N-cyclopropylcarbamimidothioate (Compound 34-2):

[0198] To a solution of cyanocarbonimidodithioic acid dimethyl ester (Compound 33-1, 1.28 g, 8.74 mmol) in ethanol (10 mL) was added cyclopropylamine (Compound 34-1, 0.5 g, 8.74 mmol). The reaction mixture was refluxed at 110 °C for 2 hours under a sealed nitrogen atmosphere. The reaction solution was cooled to room temperature, and the precipitated solid was filtered. The solid was concentrated in vacuo to obtain the crude product Compound 34-2 (1.1 g), which was directly used in the next step.

[0199] MS: (ESI, m / z): 156.1 [M+H] + , RT(min): 2.203

[0200] Step 2: Synthesis of (R,E)-N'-cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-cyclopropylpiperidine-1-carboximidamide (Compound 34):

[0201] To a solution of (E)-N'-cyano-N-cyclopropylcarbamimidothioate (Compound 34-2, 25 mg, 0.16 mmol) in N,N-dimethylformamide (2 mL) were added the hydrochloride salt of (R)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-(piperidin-3-yl)urea (Compound 1-10) (43 mg, 0.10 mmol), triethylamine (43 mg, 0.42 mmol), and silver trifluoromethanesulfonate (35 mg, 0.13 mmol). The reaction mixture was stirred at 25 °C under a nitrogen atmosphere for 1 hour. The reaction solution was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (10 mL×3). The organic phase was washed with saturated brine (10 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by high-performance liquid chromatography under the following conditions (column specification: Sunfire C18, 19*250 mm, 10 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: from 54% B to 54% B in 17 min; detection wavelength: 254 nm / 220 nm; retention time (min): 8.0 - 9.0), to obtain Compound 34 (16.41 mg).

[0202] MS: (ESI, m / z): 483.5 [M+H] + , RT(min): 1.754

[0203] 1 1H NMR (400 MHz, DMSO-d6) δ 7.48–7.39 (m, 2H), 7.33 (dd, 1H), 7.23 (d, 1H), 6.96 (t, 1H), 4.31 (d, 2H), 3.91 (d, 1H), 3.83 (d, 1H), 3.52–3.38 (m, 1H), 3.12 (t, 1H), 2.90–2.79 (m, 1H), 2.73–2.60 (m, 1H), 2.47–2.41 (m, 1H), 2.08–1.89 (m, 1H), 1.82–1.59 (m, 2H), 1.48–1.32 (m, 1H), 0.94–0.80 (m, 2H), 0.78–0.69 (m, 2H), 0.68–0.59 (m, 2H), 0.59–0.49 (m, 2H).

[0204] Example 8 (R,E)-N'-Cyano-3-(1-cyclopropyl-3-((3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)ureido)-N-methylpiperidine-1-carboxamidine (Compound 35)

[0205]

[0206] First step: Synthesis of (E)-3-(trifluoromethoxy)benzaldoxime (Compound 35-2):

[0207] Hydroxylamine hydrochloride (1.46 g, 21.04 mmol) was added to a methanol (20 mL) solution of 3-(trifluoromethoxy)benzaldehyde (Compound 35-1, 2 g, 10.52 mmol), and the mixture was reacted at 60 °C for 4 h. After the reaction was completed, water (500 mL) and dichloromethane (50 mL × 3) were added to the reaction solution for extraction. The organic phase was washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain Compound 35-2 (2.15 g).

[0208] MS: (ESI, m / z): 247.0 [M+H+41] + , RT (min): 1.732

[0209] Second step: Synthesis of tert-butyl ((3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)carbamate (Compound 35-4):

[0210] To a solution of (E)-3-(trifluoromethoxy)benzaldoxime (Compound 35-2, 2.1 g, 10.24 mmol) in methanol (20 mL) and water (5 mL) was added iodobenzene diacetate (4.29 g, 13.31 mmol) and tert-butyl prop-2-yn-1-ylcarbamate (1.59 g, 10.24 mmol), and the reaction was carried out at room temperature for 2 h. After completion of the reaction, water (500 mL) and dichloromethane (50 mL × 3) were added to the reaction solution for extraction. The organic phase was washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, and the crude product was purified by normal phase (petroleum ether:ethyl acetate = 5:1) to obtain Compound 35-4 (670 mg).

[0211] MS:(ESI,m / z):302.9[M+H-56] + ,RT(min):1.993

[0212] Synthesis of the third step (3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methanamine (Compound 35-5):

[0213] (3-(3-(Trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)carbamic acid tert-butyl ester (Compound 35-4, 170 mg, 0.47 mmol) was dissolved in a solution of hydrogen chloride in dioxane (3 mL), and the reaction was carried out at room temperature for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain Compound 35-5 (137 mg).

[0214] MS:(ESI,m / z):259.0[M+H] + ,RT(min):1.284

[0215] Synthesis of the fourth step N-((3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)-1H-imidazole-1-carboxamide (Compound 35-6):

[0216] To a solution of (3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methanamine (Compound 35-5, 137 mg, 0.46 mmol) in acetonitrile (3 mL) was added N,N'-carbonyldiimidazole (75 mg, 0.46 mmol) and triethylamine (0.2 mL, 1.38 mmol), and the reaction was carried out at 50 °C for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain Compound 35-6 (270 mg).

[0217] MS:(ESI,m / z):352.9[M+H] + ,RT(min):1.446

[0218] Step 5. Synthesis of tert-butyl (R)-3-(1-cyclopropyl-3-((3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)ureido)piperidine-1-carboxylate (Compound 35-7):

[0219] To a solution of N-((3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)-1H-imidazole-1-carboxamide (Compound 35-6, 270 mg, 0.42 mmol) in acetonitrile (3 mL) were added (3R)-3-(cyclopropylamino)piperidine-1-carboxylate tert-butyl ester (Compound 1-8, 101 mg, 0.42 mmol) and triethylamine (0.17 mL, 1.26 mmol), and the mixture was reacted at 50 °C for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain a crude product, and the crude product was purified by normal phase (petroleum ether:ethyl acetate = 3:1) to obtain Compound 35-7 (150 mg).

[0220] MS: (ESI, m / z): 525.1 [M+H] + , RT(min): 2.065

[0221] Step 6. Synthesis of (R)-1-cyclopropyl-1-(piperidin-3-yl)-3-((3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)urea (Compound 35-8):

[0222] Compound (R)-3-(1-cyclopropyl-3-((3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)ureido)piperidine-1-carboxylate tert-butyl ester (Compound 35-7, 150 mg, 0.29 mmol) was dissolved in a solution of hydrogen chloride in dioxane (3 mL), and the mixture was reacted at room temperature for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain Compound 35-8 (110 mg).

[0223] MS: (ESI, m / z): 425.0 [M+H] + , RT(min): 1.395

[0224] Step 7. Synthesis of (R,E)-N'-cyano-3-(1-cyclopropyl-3-((3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)ureido)-N-methylpiperidine-1-carboxamidine (Compound 35):

[0225] To a solution of (R)-1-cyclopropyl-1-(piperidin-3-yl)-3-((3-(3-(trifluoromethoxy)phenyl)isoxazol-5-yl)methyl)urea (Compound 35-8, 110 mg, 0.26 mmol) in N,N-dimethylformamide (3 mL) were added (E)-N'-cyano-N-methylcarbamimidothioate (Compound 3-1, 40 mg, 0.31 mmol), silver trifluoromethanesulfonate (87 mg, 0.34 mmol) and triethylamine (0.14 mL, 1.04 mmol), and the mixture was reacted at room temperature for 1 h. After completion of the reaction, water (100 mL) and dichloromethane (20 mL×3) were added to the reaction solution for extraction. The organic phase was washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was then purified by reverse-phase preparative chromatography (conditions as follows: chromatography column specifications: Prep-HPLC (Waters 2767 / QDA), Column: SunFire C18, 19*250 mm, 10 μm; mobile phase A: 0.1% formic acid / water, mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: 52% - 52%; retention time: 8.8 - 9.7 min) to obtain Compound 35 (18.4 mg).

[0226] MS: (ESI, m / z): 505.9[M+H] + , RT(min): 1.753

[0227] 1 1H NMR (400 MHz, DMSO-d6) δ 7.94 (d, 1H), 7.86 (s, 1H), 7.66 (t, 1H), 7.51 (d, 1H), 7.23–7.15 (m, 1H), 7.10 (t, 1H), 6.91 (s, 1H), 4.44 (d, 2H), 3.93 (d, 1H), 3.86 (d, 1H), 3.61–3.48 (m, 1H), 3.19 (t, 1H), 2.81 (d, 3H), 2.73 (t, 1H), 2.48–2.43 (m, 1H), 2.09–1.96 (m, 1H), 1.88–1.78 (m, 1H), 1.76–1.65 (m, 1H), 1.54–1.38 (m, 1H), 0.97–0.83 (m, 2H), 0.78–0.66 (m, 2H).

[0228] Example 9 (R,E)-N'-Cyano-3-(1-cyclopropyl-3-((6-(trifluoromethyl)naphthalen-2-yl)methyl)ureido)-N-methylpiperidine-1-carboximidamide (Compound 36)

[0229]

[0230] Step 1: Synthesis of N-((6-(trifluoromethyl)naphthalen-2-yl)methyl)-1H-imidazole-1-carboxamide (Compound 36-2):

[0231] To a solution of (6-(trifluoromethyl)naphthalen-2-yl)methanamine (Compound 36-1, 90 mg, 0.40 mmol) in acetonitrile (3 mL) were added N,N'-carbonyldiimidazole (78 mg, 0.48 mmol) and triethylamine (0.17 mL, 1.20 mmol), and the mixture was reacted at 50 °C for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain Compound 36-2 (116 mg).

[0232] MS: (ESI, m / z): 320.0 [M+H] + , RT (min): 1.463

[0233] Step 2: Synthesis of tert-butyl (R)-3-(1-cyclopropyl-3-((6-(trifluoromethyl)naphthalen-2-yl)methyl)ureido)piperidine-1-carboxylate (Compound 36-3):

[0234] To a solution of N-((6-(trifluoromethyl)naphthalen-2-yl)methyl)-1H-imidazole-1-carboxamide (Compound 36-2, 116 mg, 0.36 mmol) in acetonitrile (3 mL) were added (3R)-3-(cyclopropylamino)piperidine-1-carboxylic acid tert-butyl ester (Compound 1-8, 95 mg, 0.40 mmol) and triethylamine (0.15 mL, 1.08 mmol), and the mixture was reacted at 50 °C for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain a crude product, and the crude product was purified by normal phase chromatography (petroleum ether:ethyl acetate = 1:1) to obtain Compound 36-3 (100 mg).

[0235] MS: (ESI, m / z): 436.1 [M+H - 56] + , RT (min): 2.100

[0236] Step 3: Synthesis of (R)-1-cyclopropyl-1-(piperidin-3-yl)-3-((6-(trifluoromethyl)naphthalen-2-yl)methyl)urea (Compound 36-4):

[0237] Compound (R)-3-(1-cyclopropyl-3-((6-(trifluoromethyl)naphthalen-2-yl)methyl)ureido)piperidine-1-carboxylic acid tert-butyl ester (Compound 36-3, 60 mg, 0.12 mmol) was dissolved in a solution of hydrogen chloride in dioxane (2 mL), and the mixture was reacted at room temperature for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain a crude product of Compound 36-4 (75 mg).

[0238] MS: (ESI, m / z): 392.5 [M+H] +,RT(min):1.436

[0239] Step 4. Synthesis of (R,E)-N'-Cyano-3-(1-cyclopropyl-3-((6-(trifluoromethyl)naphthalen-2-yl)methyl)ureido)-N-methylpiperidine-1-carboximidamide (Compound 36):

[0240] To a solution of (R)-1-cyclopropyl-1-(piperidin-3-yl)-3-((6-(trifluoromethyl)naphthalen-2-yl)methyl)urea (Compound 36-4, 75 mg, 0.19 mmol) in N,N-dimethylformamide (3 mL) was added (E)-N'-cyano-N-methylcarbamimidothioate (Compound 3-1, 29 mg, 0.23 mmol), silver trifluoromethanesulfonate (63 mg, 0.25 mmol) and triethylamine (0.11 mL, 0.76 mmol), and the mixture was reacted at room temperature for 1 h. After completion of the reaction, water (100 mL) and dichloromethane (20 mL×3) were added to the reaction solution for extraction. The organic phase was washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was then purified by reversed-phase preparative chromatography (conditions as follows: column specifications: Prep-HPLC (Waters 2767 / QDA), Column: SunFire C18, 19*250 mm, 10 μm; mobile phase A: 0.1% formic acid / water, mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: 54% - 54%; retention time: 9.4 - 10.4 min) to obtain Compound 36 (20 mg).

[0241] MS: (ESI, m / z): 473.2 [M+H] + ,RT(min):1.817

[0242] 1 1H NMR (400 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.11 (t, 2H), 7.86 (s, 1H), 7.73 (dd, 1H), 7.61 (dd, 1H), 7.23–7.15 (m, 1H), 7.10 (t, 1H), 4.48 (d, 2H), 3.94 (d, 1H), 3.85 (d, 1H), 3.61–3.48 (m, 1H), 3.20 (t, 1H), 2.81 (d, 3H), 2.73 (t, 1H), 2.49–2.45 (m, 1H), 2.09–1.98 (m, 1H), 1.87–1.78 (m, 1H), 1.76–1.68 (m, 1H), 1.53–1.41 (m, 1H), 0.98–0.86 (m, 2H), 0.77–0.65 (m, 2H).

[0243] Using conditions similar to those in Example 9 above, the compounds in Table 1 below were prepared, and the structural characterization data of these compounds are listed in Table 1 together.

[0244] Table 1

[0245]

[0246]

[0247] Example 10 (R,E)-N'-Cyano-3-(1-cyclopropyl-3-(2-fluoro-4-((trifluoromethyl)thio)benzyl)ureido)-N-methylpiperidine-1-carboximidamide (Compound 37)

[0248]

[0249] First step: Synthesis of 4-bromo-3-fluorobenzenethiol (Compound 37-2):

[0250] Dissolve 4-bromo-3-fluorobenzenesulfonyl chloride (Compound 37-1, 2.0 g, 7.31 mmol) in toluene (5 mL), slowly add triphenylphosphine (2.11 g, 8.04 mmol) at room temperature, and stir at 50 °C for 15 minutes. After the reaction is completed, add water (10 mL) to the reaction solution to quench the reaction and stir for 10 minutes, then discard the aqueous layer and extract the organic layer with 10% NaOH solution (10 mL). Wash the basic extract with toluene (5 mL), acidify with dilute HCl and extract with dichloromethane (15 mL × 3). Combine the organic phases, wash with saturated brine (20 mL × 2), dry over anhydrous sodium sulfate, filter and concentrate to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to obtain Compound 37-2 (1.2 g).

[0251] MS: (ESI, m / z): 205.0 [M-H] - , RT(min): 0.910

[0252] Second step: Synthesis of (4-bromo-3-fluorophenyl)(trifluoromethyl) sulfane (Compound 37-4):

[0253] 4-Bromo-3-fluorobenzenethiol (Compound 37-2, 1.0 g, 4.83 mmol) was dissolved in acetonitrile (12 mL). Triethylamine (0.54 g, 5.31 mmol) was added to lower the temperature of the reaction solution to -20 °C. Trifluoromethyl iodide (Compound 37-3, 4.16 g, 5.31 mmol) and sodium metabisulfite (0.92 g, 4.83 mmol) were added. The reactants were heated to room temperature and stirred overnight. Water (50 mL) was added to dilute the reaction solution, and it was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude target compound. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 500:1) to obtain Compound 37-4 (1.1 g).

[0254] 1 H NMR (400 MHz, CDCl3) δ 7.59–7.52 (m, 1H), 7.36 (dd, 1H), 7.26 (d, 1H).

[0255] Step 3: Synthesis of tert-butyl (2-fluoro-4-((trifluoromethyl)thio)benzyl)carbamate (Compound 37-5):

[0256] (4-Bromo-3-fluorophenyl)(trifluoromethyl) sulfane (Compound 37-4, 1.1 g, 4.00 mmol) was dissolved in dioxane (10 mL) and water (1.25 mL). At room temperature, potassium ((tert-butoxycarbonyl)amino)methyl)trifluoroborate (1.04 g, 4.40 mmol), potassium carbonate (1.66 g, 12.0 mmol), palladium acetate (36 mg, 0.16 mmol), and 2-(dicyclohexylphosphino)-2,4,6-triisopropylbiphenyl (150 mg, 0.32 mmol) were added. The gas was displaced. Under nitrogen protection, the reaction solution was stirred at room temperature for 20 minutes and then at 110 °C overnight. The reaction was cooled to room temperature, filtered, water (30 mL) was added to dilute the reaction solution, and it was extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain Compound 37-5 (650 mg).

[0257] MS: (ESI, m / z): 326.1 [M+H] + , RT(min): 0.999

[0258] Step 4: Synthesis of (2-fluoro-4-((trifluoromethyl)thio)phenyl)methanamine (Compound 37-6):

[0259] To a solution of tert-butyl (2-fluoro-4-((trifluoromethyl)thio)benzyl)carbamate (Compound 37-5, 650 mg, 2.00 mmol) in dioxane (3 mL) was added hydrogen chloride - dioxane solution (3 mL, 4 M), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated to obtain crude Compound 37-6 (440 mg).

[0260] MS: (ESI, m / z): 226.0 [M+H] + , RT(min): 1.240

[0261] Step 5. Synthesis of (R)-tert-butyl 3-(1-cyclopropyl-3-(2-fluoro-4-((trifluoromethyl)thio)benzyl)ureido)piperidine-1-carboxylate (Compound 37-7):

[0262] (2-Fluoro-4-((trifluoromethyl)thio)phenyl)methanamine (Compound 37-6, 200 mg, 0.89 mmol) was dissolved in acetonitrile (4 mL), and N,N'-carbonyldiimidazole (158.74 mg, 0.98 mmol) and triethylamine (270.18 mg, 2.67 mmol) were added. After addition, the mixture was stirred at 50 °C for 2 hours. The reaction was monitored by TLC until completion, and the reaction mixture was quenched with water (30 mL) and extracted with ethyl acetate (30 mL×3). After concentration, crude N-(2-fluoro-4-((trifluoromethyl)thio)benzyl)-1H-imidazole-1-carboxamide was obtained. (3R)-tert-Butyl 3-(cyclopropylamino)piperidine-1-carboxylate (Compound 1-8, 211 mg, 0.88 mmol) was dissolved in acetonitrile (4 mL), and the crude N-(2-fluoro-4-((trifluoromethyl)thio)benzyl)-1H-imidazole-1-carboxamide and triethylamine (267.14 mg, 2.64 mmol) were added. After addition, the mixture was stirred at 50 °C for 2 hours. Water (20 mL) was added to dilute the reaction mixture, and it was extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative normal-phase column (EA\PE = 1\3) to obtain Compound 37-7 (491.54 mg).

[0263] MS: (ESI, m / z): 492.2 [M+H] + , RT(min): 2.128.

[0264] Step 6. Synthesis of (R)-1-cyclopropyl-3-(2-fluoro-4-((trifluoromethyl)thio)benzyl)-1-(piperidin-3-yl)urea (Compound 37-8):

[0265] To a solution of tert-butyl (R)-3-(1-cyclopropyl-3-(2-fluoro-4-((trifluoromethyl)thio)benzyl)ureido)piperidine-1-carboxylate (Compound 37-7, 300 mg, 0.61 mmol) in dioxane (2 mL) was added hydrogen chloride-dioxane solution (2 mL, 4 M), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated to obtain crude Compound 37-8 (200 mg).

[0266] MS: (ESI, m / z): 392.1 [M+H] + , RT(min): 1.240

[0267] Synthesis of step 7, (R,E)-N'-cyano-3-(1-cyclopropyl-3-(2-fluoro-4-((trifluoromethyl)thio)benzyl)ureido)-N-methylpiperidine-1-carboximidamide (Compound 37):

[0268] At room temperature, (E)-N'-cyano-N-methylcarbamimidothioate (Compound 3-1, 71.69 mg, 0.55 mmol) and triethylamine (149.76 mg, 1.48 mmol) were added to a stirred solution of (R)-1-cyclopropyl-3-(2-fluoro-4-((trifluoromethyl)thio)benzyl)-1-(piperidin-3-yl)urea (Compound 37-8, 160 mg, 0.37 mmol) in N,N-dimethylformamide (3 mL). Finally, silver trifluoromethanesulfonate (123.59 mg, 0.48 mmol) was slowly added. The reaction mixture was stirred at room temperature for 1 hour. Water (20 mL) was added to the reaction mixture to dilute it, and it was extracted with dichloromethane (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated. The conditions were as follows (column chromatography specifications: Sunfire C18, 19*250 mm, 10 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: from 32% B to 37% B in 17 min; detection wavelength: 254 nm / 220 nm; retention time (min): 8.1 - 10.2), to obtain Compound 37 (27.93 mg).

[0269] MS: (ESI, m / z): 472.8 [M+H] + , RT(min): 1.745

[0270] 11H NMR (400 MHz, DMSO-d6) δ 7.62–7.54 (m, 2H), 7.45 (t, 1H), 7.22–7.13 (m, 1H), 7.02 (t, 1H), 4.36 (d, 2H), 3.96–3.80 (m, 2H), 3.55–3.48 (m, 1H), 3.19 (t, 1H), 2.81 (d, 3H), 2.76–2.68 (m, 1H), 2.49–2.46 (m, 1H), 2.07–1.94 (m, 1H), 1.84–1.67 (m, 2H), 1.52–1.41 (m, 1H), 0.94–0.86 (m, 2H), 0.73–0.66 (m, 2H).

[0271] Using conditions similar to those in Example 10 above, the compounds in Table 2 below were prepared, and the structural characterization data of these compounds are listed in Table 2 together.

[0272] Table 2

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281] Example 11 (E)-N'-Cyano-4-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-methyl-2-azabicyclo[4.1.0]heptane-2-carboximidamide (Compound 83)

[0282]

[0283] Note: There is chirality, that is, 83-4-P1 and 83-4-P2 are each either of them, and 83-4-P1 and 83-4-P2 are different.

[0284] There is chirality, that is, 83-6-P1 is a mixture of or mixture

[0285] Chirality exists, that is, 83-P1 is mixture or mixture

[0286] First step: Synthesis of benzyl 3-hydroxy-3,4-dihydropyridine-1(2H)-carboxylate (Compound 83-2):

[0287] At -78 °C, sodium borohydride (17 g, 450 mmol) was added to a methanol (400 mL) solution of pyridin-3-ol (Compound 25-1, 20 g, 210 mmol), and added slowly while maintaining the temperature below -60 °C. Sodium bicarbonate (13 g, 156 mmol) and benzyl chloroformate (53 g, 309 mmol) were successively added to the reaction solution, and the temperature was gradually raised to room temperature and stirred overnight. The reaction was quenched with 1N aqueous sodium hydroxide solution (100 mL), then water (300 mL) was added to the reaction solution, and it was extracted with ethyl acetate (300 mL × 3). The organic phases were combined, washed with saturated brine (300 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to obtain Compound 83-2 (20 g).

[0288] MS: (ESI, m / z): 234.1 [M+H] + , RT(min): 9.337

[0289] Second step: Synthesis of benzyl 4-hydroxy-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-3):

[0290] At room temperature, diethylzinc (85.76 mL, 85.76 mmol) and diiodomethane (31 g, 11.63 mmol) were successively added to a 1,2-dichloroethane (40 mL) solution of benzyl 3-hydroxy-3,4-dihydropyridine-1(2H)-carboxylate (Compound 83-2, 5 g, 21.44 mmol). The reaction system was stirred at room temperature for 1 h. The reaction was quenched by adding saturated sodium bicarbonate (50 mL), then the reaction solution was diluted with water (200 mL), extracted with ethyl acetate (200 mL × 3). The organic phases were combined, washed with saturated brine (200 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain Compound 83-3 (2.6 g).

[0291] MS: (ESI, m / z): 248.0 [M+H]+ ,RT(min): 1.595

[0292] Synthesis of benzyl 4-oxo-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-4) in the third step:

[0293] At -78 °C, dimethyl sulfoxide (1.97 g, 25.22 mmol) was added to a solution of oxalyl chloride (1.60 g, 12.61 mmol) in dichloromethane (25 mL), and the mixture was stirred for 15 min. Then benzyl 4-hydroxy-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-3, 2.60 g, 10.51 mmol) was added, and the mixture was stirred for 35 min. Then triethylamine (5.32 g, 52.55 mmol) was added dropwise, and the reaction was carried out at -78 °C for 10 min and then continued at room temperature for 1 h. Water (150 mL) was added to the reaction solution to quench the reaction, and the mixture was extracted with ethyl acetate (150 mL × 3). The organic phases were combined, washed with saturated brine (150 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain Compound 25-4 (2.2 g). After chiral SFC separation, the conditions were as follows (column specification: AD 250*25 mm 10 μm; mobile phase A: supercritical CO2, mobile phase B: EtOH (+0.1% EtOH solution of 7.0 mol / L ammonia); flow rate: 120 mL / min; elution gradient: isocratic 80; detection wavelength: UV 214 nm; retention time of the front peak (min): 4.15; retention time of the rear peak (min): 11.55; sample dissolution solvent: EtOH; single injection volume: 8.0 mL), to obtain Compound 83-4-P1 (front peak, 700 mg) and Compound 83-4-P2 (rear peak, 750 mg).

[0294] 83-4-P1: MS: (ESI, m / z): 246.1 [M+H] + ,RT(min): 10.158

[0295] 83-4-P2: MS: (ESI, m / z): 246.1 [M+H] + ,RT(min): 10.289

[0296] Synthesis of benzyl 4-(cyclopropylamino)-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-6-P1) in the fourth step:

[0297] At room temperature, to a solution of benzyl 4-oxo-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-4-P1, 650 mg, 2.65 mmol) in ethanol (10 mL), cyclopropylamine (Compound 25-5, 605 mg, 10.60 mmol) and titanium(IV) isopropoxide (1.5 g, 5.30 mmol) were added. The mixture was stirred at room temperature for 2 hours, then sodium borohydride (301 mg, 7.95 mmol) was added to the reaction system and stirred at room temperature for another 2 hours. Water (70 mL) was added to the reaction solution to quench the reaction, and the mixture was filtered. The filtrate was extracted with ethyl acetate (70 mL×3), and the combined organic phases were washed with saturated brine (70 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain Compound 83-6-P1 (300 mg).

[0298] MS:(ESI,m / z):287.1[M+H] + ,RT(min):6.760

[0299] Synthesis of benzyl 4-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-7-P1) in the fifth step:

[0300] To a solution of N-(2-fluoro-4-(trifluoromethoxy)benzyl)-1H-imidazole-1-carboxamide (Compound 1-5, 277 mg, 0.91 mmol) in acetonitrile (5 mL), benzyl 4-(cyclopropylamino)-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-6-P1, 250 mg, 0.87 mmol) and triethylamine (264 mg, 2.61 mmol) were added respectively. The reaction system was stirred at 50 °C for 1 h. Water (50 mL) was added to dilute the reaction solution, and the mixture was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain Compound 83-7-P1 (300 mg).

[0301] MS:(ESI,m / z):522.2[M+H] + ,RT(min):12.804

[0302] Synthesis of 1-(2-azabicyclo[4.1.0]heptan-4-yl)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)urea (Compound 83-8-P1) in the sixth step:

[0303] To a solution of benzyl 4-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-2-azabicyclo[4.1.0]heptane-2-carboxylate (Compound 83-7-P1, 486 mg, 0.93 mmol) in tetrahydrofuran (10 mL) was added palladium on carbon (99 mg, 0.09 mmol). The hydrogen was displaced three times, and the reaction system was stirred at room temperature for 1 h. The reaction solution was filtered and concentrated to obtain the crude product Compound 83-8-P1 (300 mg), which was directly used in the next step of the reaction.

[0304] MS:(ESI,m / z):388.2[M+H] + ,RT(min):7.772

[0305] Synthesis of step 7: (E)-N'-Cyano-4-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-N-methyl-2-azabicyclo[4.1.0]heptane-2-carboximidamide (Compound 83-P1):

[0306] To a solution of 1-(2-azabicyclo[4.1.0]heptan-4-yl)-1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)urea (Compound 83-8-P1, 27 mg, 0.07 mmol) in N,N-dimethylformamide (2 mL) were added methyl N'-cyano-N-methylcarbamimidothioate (Compound 83-9, 9.04 mg, 0.07 mmol), triethylamine (28.33 mg, 0.28 mmol) and silver trifluoromethanesulfonate (26.98 mg, 0.11 mmol). The reaction was carried out at room temperature for 1 h. Water (20 mL) was added to the reaction solution for dilution, and the mixture was extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by high performance liquid chromatography under the following conditions (column specification: Sunfire C18, 19*250 mm, 10 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: from 52% B to 55% B within 16 min; detection wavelength: 254 nm / 220 nm; retention time (min): 7.60 - 8.50), to obtain Compound 83-P1 (9.47 mg).

[0307] MS:(ESI,m / z):469.3[M+H] + ,RT(min):10.592

[0308] MS:(ESI,m / z):469.3[M+H] + ,RT(min):10.666

[0309] 1 1H NMR (400 MHz, DMSO-d6) δ 7.45–7.39 (m, 1H), 7.36–7.30 (m, 1H), 7.25–7.21 (m, 1H), 7.16–7.03 (m, 1H), 7.02–6.93 (m, 1H), 4.31 (d, 2H), 3.92–3.74 (m, 1H), 3.42–3.33 (m, 2H), 2.93 (t, 3H), 2.75–2.58 (m, 1H), 2.45–2.33 (m, 1H), 2.20–1.92 (m, 1H), 1.58–1.25 (m, 2H), 0.96–0.84 (m, 3H), 0.70–0.58 (m, 2H), 0.38–0.26 (m, 1H).

[0310] Example 12 (3R,5S,E)-N'-Cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-5-hydroxy-N-methylpiperidine-1-carboximidamide (Compound 84)

[0311]

[0312] First step: Synthesis of tert-butyl (3R,5S)-3-(cyclopropylamino)-5-hydroxypiperidine-1-carboxylate (Compound 84-2):

[0313] To a solution of tert-butyl (3R,5S)-3-amino-5-hydroxypiperidine-1-carboxylate (Compound 84-1, 100 mg, 0.46 mmol) in 1,4-dioxane (2 mL) was added (1-ethoxycyclopropoxy)trimethylsilane (Compound 1-7, 40 mg, 0.23 mmol), sodium cyanoborohydride (58 mg, 0.92 mmol) and acetic acid (55 mg, 0.92 mmol). The reaction was carried out at 60 °C for 12 h. After completion of the reaction, saturated aqueous sodium bicarbonate solution was added to adjust the system to pH = 9. Dichloromethane (20 mL × 3) was added for extraction. The organic phase was washed with saturated brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by normal phase chromatography (dichloromethane:methanol = 10:1) to obtain Compound 84-2 (110 mg).

[0314] MS: (ESI, m / z): 257.0 [M+H] + , RT(min): 1.213

[0315] Second step: Synthesis of tert-butyl (3R,5S)-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-5-hydroxypiperidine-1-carboxylate (Compound 84-3):

[0316] To a solution of N-(2-fluoro-4-(trifluoromethoxy)benzyl)-1H-imidazole-1-carboxamide (Compound 1-5, 49 mg, 0.16 mmol) in acetonitrile (3 mL) were added (3R,5S)-3-(cyclopropylamino)-5-hydroxypiperidine-1-carboxylic acid tert-butyl ester (Compound 84-2, 50 mg, 0.16 mmol) and triethylamine (0.07 mL, 0.48 mmol), and the reaction was carried out at 50 °C for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain a crude product, and the crude product was purified by normal phase (petroleum ether:ethyl acetate = 1:1) to obtain Compound 84-3 (20 mg).

[0317] MS: (ESI, m / z): 436.2 [M+H-56] + , RT(min): 1.897

[0318] Synthesis of the third step, 1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-((3R,5S)-5-hydroxypiperidin-3-yl)urea (Compound 84-4):

[0319] (3R,5S)-3-(1-Cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-5-hydroxypiperidine-1-carboxylic acid tert-butyl ester (Compound 84-3, 20 mg, 0.041 mmol) was dissolved in a solution of hydrogen chloride in dioxane (1 mL), and the reaction was carried out overnight at room temperature. After completion of the reaction, the reaction solution was concentrated to obtain Compound 84-4 (15 mg).

[0320] MS: (ESI, m / z): 392.1 [M+H] + , RT(min): 1.364

[0321] Synthesis of the fourth step, (3R,5S,E)-N'-cyano-3-(1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)ureido)-5-hydroxy-N-methylpiperidine-1-carboxamidine (Compound 84):

[0322] To a solution of 1-cyclopropyl-3-(2-fluoro-4-(trifluoromethoxy)benzyl)-1-((3R,5S)-5-hydroxypiperidin-3-yl)urea (Compound 84-4, 15 mg, 0.38 mmol) in N,N-dimethylformamide (2 mL) was added N'-cyano-N-methylcarbamimidothioate (Compound 3-1, 6 mg, 0.046 mmol), silver trifluoromethanesulfonate (13 mg, 0.049 mmol) and triethylamine (0.02 mL, 0.15 mmol), and the reaction was carried out at room temperature for 1 h. After the reaction was completed, water (100 mL) and dichloromethane (20 mL × 3) were added to the reaction solution for extraction. The organic phase was washed with saturated brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was then purified by reverse-phase preparative chromatography (conditions are as follows: chromatography column specification: Prep-HPLC (Waters 2767 / QDA), Column: SunFire C18, 19*250 mm, 10 μm; mobile phase A: 0.1% formic acid / water, mobile phase B: acetonitrile; flow rate: 20 mL / min; elution gradient: 54% - 54%; retention time: 9.4 - 10.4 min) to obtain Compound 84 (3.52 mg).

[0323] MS:(ESI,m / z):473.2[M+H] + ,RT(min):1.662

[0324] 1 1H NMR(400MHz,DMSO-d6)δ7.42(t,1H),7.38–7.30(m,1H),7.30–7.17(m,2H),6.99(t,1H),5.09(d,1H),4.32(d,2H),4.07–3.95(m,1H),3.81–3.69(m,1H),3.57–3.46(m,2H),3.10(t,1H),2.82(d,3H),2.47–2.43(m,2H),2.06–1.84(m,2H),0.96–0.81(m,2H),0.74–0.60(m,2H).

[0325] Biological evaluation

[0326] Test Example 1

[0327] Test name: SLC6A19 Isoleucine Transport Assay

[0328] Cell line generation and maintenance:

[0329] The Flp-In™-CHO™ cell line was purchased from Thermo Fisher Scientific. Stable cell lines were generated by transfecting plasmids encoding TMEM27 and hSLC6A19 using standard protocols followed by antibiotic selection. The cell lines were used to generate inducible cell lines expressing TMEM27 and hSLC6A19. The stable cells were maintained in DMEM / F12 (Thermo Fisher) supplemented with 10% fetal bovine serum, 800 μg / ml G418, and 2 mM L-glutamine.

[0330] Buffer preparation:

[0331] 1. Component A (FLIPR Membrane Potential Assay Kit)

[0332] 2. Component B: 20 mM HEPES dissolved in 1× HBSS buffer, adjusted to pH 7.4 with 1 N NaOH.

[0333] Compound preparation:

[0334] 1. Dissolve the compound in DMSO to make a 10 mM stock solution, aliquot and store at -20 °C.

[0335] 2. Dilute the compound with buffer to the highest test concentration of 10000 nM, with 2-fold dilutions.

[0336] a) The final test concentrations of JNT-517 were: 10000, 5000, 2500, 1250, 625, 312.5, 156.3, 78.1, 39.1, 19.5 nM.

[0337] b) For the test compound, the final test concentrations were: 10000, 5000, 2500, 1250, 625, 312.5, 156.3, 78.1, 39.1, 19.5 nM.

[0338] Seed the cells into a 384-well cell culture plate

[0339] 1. Culture TMEM27 / hSLC6A19 CHO cells in cell culture medium (F12, 10% FBS, 800 μg / ml G418, 2 mM L-glutamine).

[0340] 2. When the cell confluence reaches 80%, dissociate the cells with 0.25% trypsin. Measure the cell density and dilute the cells to 1.0×10 6 cells / mL with the medium.

[0341] Add 20 μL of cells to each well of a 384-well black experimental plate (pre-coated with poly-D-lysine or matrix), and incubate overnight at 37 °C in 5% CO₂.

[0342] Detection:

[0343] 1. On the day of the experiment, wash several times with Component B to completely dissolve the contents of the Component A vial, making the total volume 10 mL.

[0344] 2. Gently centrifuge to remove the cell plate medium, add 20 μL of Component B and 20 μL of buffer to each well, and incubate at room temperature for 30 minutes.

[0345] 3. After incubation, add 10 μL of a 5-fold final concentration of the compound to the detection plate, and add 10 μL of a 6-fold final concentration of L-isoleucine to a final concentration of 10 mM. Incubate the plate for 15 minutes. Read the fluorescence value using FLIPR (excitation / emission wavelength 510 - 545 / 565 - 625, 5 minutes).

[0346] Data analysis:

[0347] Inhibition rate (%) = 100 - (reading of the compound well - reading of the low-reading control well) / (reading of the high-reading control well - reading of the low-reading control well) × 100

[0348] High-reading control well: 0.1% DMSO; Low-reading control well: 10 μM JNT-517.

[0349] Use GraphPad Prism 9 software to calculate IC 50 (nM) and plot the effect-dose curve of the compound.

[0350] Table 3 Biological activity data of the compounds of the present application

[0351]

[0352]

[0353] The above has given an exemplary description of the implementation manner of the technical solution of the present invention. It should be understood that the protection scope of the present invention is not limited to the above implementation manner. Any modifications, equivalent replacements, improvements, etc. made by those skilled in the art within the spirit and principle of the present invention shall be included within the protection scope of the claims of this application.

Claims

1. A compound represented by formula (I), its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound: in, R1 is selected from H, CN, -OH, unsubstituted or optionally substituted with one, two or more R d Substituted with the following groups: C 1-12 Alkyl, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl; each R d are the same or different and are independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy or halogenated C 1-6 alkyl; R2 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: -N(R 21 )(R 22 ), C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl or 3-14 membered heterocyclic group; R 21 , R 22 The same or different, independently selected from H, C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, C 3-12 Cycloalkyl, halogenated C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 aryl or 5-10 membered heteroaryl; or, R 21 , R 22 and the N atom to which it is attached form a 3-14 membered heterocyclic ring; each R b are the same or different and are independently selected from OH, CN, halogen, oxo (=O), unsubstituted or optionally substituted by one, two or more R b1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -NH2 or -S(O)2-C 1-12 Alkyl; each R b1 the same or different, independently selected from OH, NH2, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy or halogenated C 1-6 alkyl; R3 and R4 are the same or different and are independently selected from H, halogen, unsubstituted or optionally substituted by one, two or more selected from halogen, CN, OH, C 1-12 Alkyl, halogenated C 1-12 The following groups substituted by the alkyl group: C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, C 3-12 Cycloalkyl, halogenated C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl; L1 is absent or selected from unsubstituted or optionally substituted with one, two or more R e Substituted with the following groups: C 1-12 Alkylene, C 2-6 Alkenylene, C 2-6 Alkynylidene, C 3-14 Cycloalkylene, C 6-14 Arylene, 5-14 membered heteroarylene, C 6-14 Arylene-C 1-12 Alkylene or 5-14 membered heteroarylene-C 1-12 Alkylene; each R e are the same or different and are independently selected from H, OH, CN, halogen, NH2, C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, C 3-12 Cycloalkyl, halogenated C 3-12 Cycloalkyl; L2 does not exist or is selected from C 1-12 Alkylene, C 2-6 Alkenylene, C 2-6 Alkynylidene, C 3-14 Cycloalkylene, C 6-10 Arylene or 5-10 membered heteroarylene; Y1 is selected from unsubstituted or optionally substituted with one, two or more R c Substituted with the following groups: C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl; each R c are the same or different and are independently selected from H, OH, CN, halogen, unsubstituted or optionally substituted by one, two or more R c1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 1-12 Alkylthio, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -NH2 or -S(O)2-C 1-12 Alkyl; each R c1 are the same or different and are independently selected from H, OH, NH2, CN, halogen, C 1-6 Alkyl or C 1-6 Alkoxy; Each R a are the same or different and are independently selected from H, CN, oxo (=O), halogen, OH, unsubstituted or optionally substituted by one, two or more R a1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -C(O)N(R a11 )(R a12 )、-N(R a13 )(R a14 )、-S(O)2-R a15 、-S(O)(=NR a16 )(R a17 ) or -P(O)(R a18 )(R a19 ); or, two R attached to the same carbon atom a Together with the carbon atom to which it is attached, it forms an unsubstituted or optionally substituted with one, two or more R a1 Substituted ring system: C 3-14 A carbocyclic ring or a 3-14 membered heterocyclic ring; or two R a Together with the carbon atoms to which they are attached, they are unsubstituted or optionally substituted with one, two or more R a1 Substituted ring system: C 3-14 Carbocyclic ring, 3-14 membered heterocyclic ring, C 6-14 aromatic ring or 5-14 membered heteroaromatic ring; or, two non-adjacent R a connected with their end groups to form an unsubstituted or optionally substituted R a1 Substituted C 1-3 alkylene; each R a1 are the same or different and are independently selected from H, OH, CN, halogen, unsubstituted or optionally substituted by one, two or more R a2 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -NH2 or -S(O)2-C 1-12 Alkyl; each R a2 are the same or different and are independently selected from H, OH, NH2, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy or halogenated C 1-6 Alkyl; R a11 , R a12 , R a13 , R a14 , R a15 , R a16 , R a17 , R a18 , R a19 The same or different, independently selected from H, C 1-12 Alkyl, C 3-8 Cycloalkyl or 3-8 membered heterocyclic group; m is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8; n is selected from 0, 1, 2 or 3.

2. The compound according to claim 1, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound, characterized in that: Each R a are the same or different and are independently selected from H, CN, oxo (=O), halogen, OH, unsubstituted or optionally substituted by one, two or more R a1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -C(O)N(R a11 )(R a12 )、-N(R a13 )(R a14 )、-S(O)2-R a15 、-S(O)(=NR a16 )(R a17 ) or -P(O)(R a18 )(R a19 ); each R a1 are the same or different and are independently selected from H, OH, CN, halogen, unsubstituted or optionally substituted by one, two or more R a2 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -NH2 or -S(O)2-C 1-12 Alkyl; each R a2 are the same or different and are independently selected from H, OH, NH2, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy or halogenated C 1-6 Alkyl; R a11 , R a12 , R a13 , R a14 , R a15 , R a16 , R a17 , R a18 , R a19 The same or different, independently selected from H, C 1-12 Alkyl, C 3-8 Cycloalkyl or 3-8 membered heterocyclic group; Preferably, m is selected from 0, 1 or 2; Preferably, each R a are the same or different and are independently selected from CN, oxo (=O), halogen (e.g. F, Cl, Br), OH, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; or, two R connected to adjacent carbon atoms a Together with the carbon atoms to which they are attached, they form C 3-6 Carbon ring; Preferably, R a is OH; or, two R a together with the carbon atoms to which they are respectively attached, form a cyclopropyl ring; R1 is selected from H, CN, -OH, unsubstituted or optionally substituted with one or two R d Substituted with the following groups: C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclyl, phenyl or 5-6 membered heteroaryl; each R d The same or different, independently selected from C 1-3 alkyl; Preferably, R1 is selected from H, CN, -OH, C 1-4 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl), C 3-6 Cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl), 4-6 membered heterocyclic group (e.g., ), phenyl or 5-6 membered heteroaryl (e.g. ); Preferably, R1 is selected from H, CN, -OH, methyl, ethyl, cyclopropyl, cyclopentyl, Preferably, R1 is selected from CN or OH.

3. The compound according to claim 1 or 2, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, characterized in that: R2 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: -N(R 21 )(R 22 ), C 1-12 Alkyl, C 3-12 Cycloalkyl or 3-14 membered heterocyclic group; Preferably, R2 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: -N(R 21 )(R 22 ), C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl), C 3-6 Cycloalkyl (e.g. cyclopropyl) or 4-6 membered heterocyclic group (e.g. ); Preferably, R 21 , R 22 The same or different, independently selected from H, C 1-12 Alkyl, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 aryl or 5-10 membered heteroaryl; or, R 21 , R 22 Together with the N atom to which it is attached, it forms a 3-14 membered heterocyclic ring; Preferably, R 21 , R 22 are the same or different and are independently selected from H, methyl, ethyl, cyclopropyl, Or, R 21 , R 22 and the N atom to which it is attached Preferably, R 21 , R 22 The same or different, independently selected from H, C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl), Or, R 21 , R 22 and the N atom to which it is attached Preferably, R 21 Selected from H, R 22 Selected from C 1-6 Alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl); Preferably, R2 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: methyl, ethyl, cyclopropyl, -NH2, -NHCH3, -NHCH2CH3, Preferably, R2 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: methyl, ethyl, cyclopropyl, -NH2, -NHCH3, Preferably, each R b are the same or different and are independently selected from OH, CN, halogen or C 1-12 alkyl; Preferably, each R b The same or different, independently selected from OH, CN, F, C 1-6 Alkyl (eg, methyl, ethyl, n-propyl, isopropyl, n-butyl or tert-butyl).

4. The compound according to any one of claims 1 to 3, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, characterized in that: L2 does not exist; Preferably, R3 and R4 are the same or different and are independently selected from H, halogen, C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, C 3-12 Cycloalkyl, halogenated C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl; Preferably, R3 is selected from H; Preferably, R4 is selected from unsubstituted or optionally substituted by a halogen (such as F, Cl, Br) or C 1-4 Cyclopropyl substituted with an alkyl group (e.g., methyl, ethyl, isopropyl, tert-butyl); Preferably, R4 is selected from Preferably, R4 is selected from cyclopropyl; Preferably, L1 is selected from C 1-4 Alkyl (such as methyl, ethyl, isopropyl, tert-butyl) or OH substituted C 1-6 Alkylene or 5-6 membered heteroarylene-C 1-6 Alkylene; Preferably, L1 is selected from methylene, -CH(CH3)-, -CH(CH2OH)-, or The "*" side is connected to Y1, and the "#" side is connected to N; Preferably, L1 is selected from methylene; Preferably, Y1 is selected from unsubstituted or optionally substituted with one, two or more R c substituted phenyl, naphthyl or 5-6 membered heteroaryl; each R c are the same or different and are independently selected from H, CN, halogen, unsubstituted or optionally substituted by one, two or more R c1 Substituted with the following groups: C 1-6 Alkyl, C 1-6 Alkoxy or C 1-6 Alkylthio; each R c1 are the same or different and are independently selected from H, CN or halogen; Preferably, Y1 is selected from unsubstituted or optionally substituted with one, two or more R c Substituted phenyl or 5-6 membered heteroaryl; each R c are the same or different and are independently selected from H, CN, halogen, unsubstituted or optionally substituted by one, two or more R c1 Substituted with the following groups: C 1-6 Alkyl or C 1-6 Alkoxy; each R c1 are the same or different and are independently selected from H, CN or halogen; Preferably, Y1 is selected from unsubstituted or optionally substituted with one, two or three R c Substituted phenyl, naphthyl, pyridyl (such as ), pyrazolyl (such as ), thiazolyl (such as ) or thienyl (such as ); Preferably, each R c The same or different, independently selected from F, Cl, Br, CN, CH3, CF3, CH2CF3, CF2CF3, OCH3, OCF3, OCF2H, OCF2Br, OCF2Cl or SCF3; Preferably, Y1 is selected from Preferably, Y1 is selected from 5. The compound according to any one of claims 1 to 4, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, characterized in that: The compound represented by formula (I) has the structure shown below: Among them, R1, R2, R3, R4, Y1, L1, R c Having the definition of any one of claims 1 to 4; Preferably, the compound represented by formula (I) has the structure shown below: Wherein, R1 and R2 have the definitions as described in any one of claims 1 to 4; Preferably, the compound represented by formula (I) has the structure shown below: Among them, Y1, L1, R3, R4, R 21 , R 22 It has the definition as described in any one of claims 1 to 4.

6. The compound according to any one of claims 1 to 5, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, characterized in that: The compound represented by formula (I) is selected from the following structures:

7. A method for preparing a compound of formula (I) according to any one of claims 1 to 6, comprising the following steps A: in, R1, R2, R3, R4, R a , L1, L2, Y1, m, n have the definitions as described in any one of claims 1-4.

8. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound of formula (I) according to any one of claims 1 to 6, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof; Preferably, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients; Preferably, the pharmaceutical composition further contains one or more additional therapeutic agents.

9. A method for treating or preventing a disease or condition mediated by SLC6A19, comprising administering to a patient a preventive or therapeutically effective amount of at least one of the compound of formula (I) according to any one of claims 1 to 6, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, or the pharmaceutical composition according to claim 8; Preferably, the SLC6A19-mediated disease or disorder is phenylketonuria; and the patient comprises a mammal, preferably a human.

10. Use of the compound of formula (I) according to any one of claims 1 to 6, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, or the pharmaceutical composition according to claim 8 in the preparation of a drug; Preferably, the use may be for preparing a drug for treating or preventing a disease or condition mediated by SLC6A19; Preferably, the SLC6A19-mediated disease or disorder is phenylketonuria.

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