Tetrazole compound, pharmaceutical composition thereof and use thereof

Abstract

The present invention provides a tetrazole compound, a pharmaceutical composition thereof and a use thereof. The compound of the present invention has a brand-new structure, a low clearance rate, high plasma exposure, good oral bioavailability, and good pharmacokinetic properties, and is conducive to drug development. The compound of the present invention or a pharmaceutically acceptable salt thereof can be used for preventing and / or treating pain, including neuropathic pain, etc.

Description

Tetrazol-containing compounds, pharmaceutical compositions and uses thereof

[0001] This application claims priority to Chinese patent applications CN2024102282843, filed on February 29, 2024, CN2024105084581, filed on April 26, 2024, CN2024110191507, filed on July 29, 2024, CN2024114236812, filed on October 12, 2024, and CN202411887462X, filed on December 20, 2024. The entire text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] The present invention belongs to the field of medicine, and in particular relates to a tetrazole compound, a pharmaceutical composition thereof and use thereof. Background Art

[0003] The renin-angiotensin system (RAS) is a crucial fluid regulation system in the human body, with angiotensin II (Ang II) as its primary component. It not only plays a crucial role in maintaining circulatory homeostasis and water and sodium retention but also participates in other physiological functions, including the growth and development of the nervous system. Accumulating evidence indicates that the RAS plays a crucial role in neuroprotection and neuroregeneration, and studies have shown that the Ang II metabolic pathway is associated with pain sensitivity in humans. Ang II receptors primarily include type 1 (AT1R) and type 2 (AT2R), both G protein-coupled receptors with similar affinity for Ang II. AT2R, a G protein-coupled receptor, is implicated in pain mechanisms in the nervous system and is primarily expressed in the dorsal and trigeminal ganglia. Compared to normal nerves, damaged nerves and painful neuromas have higher AT2R expression. Activated AT2R sensitizes ion channels in neurons through G protein-coupled receptor-activated second messenger pathways. Sensitization leads to ion channel activation, which in turn excites neurons. AT2R antagonists have been shown to be effective in relieving pain in animal studies (Pain. Medicine. 2013, 14, 1557-1568; Pain. Medicine. 2013, 14, 692-705) and clinical trials (Lancet. 2014, 383, 1637-1647). Related reviews can be found in Expert Opin. Investig. Drugs. 2014, 23, 1-12; Expert. Opin. Ther. Targets. 2015, 19, 25-35. Summary of the Invention

[0004] The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0005] Wherein: M1 and M2 are each independently selected from C or N;

[0006] L1 is a bond or -CH2-;

[0007] L2 is a bond, -CH2-, -C(O)-, -NH-, -C(O)NH-, -NHC(O)-, -O- or -S-, wherein -CH2- and -NH- are optionally replaced by one or two R L2 Replacement, R L2 Each independently selected from halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C1-3 Halogenated alkyl, C 1-3 Halogenated alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group;

[0008] L is selected from L3 or ring A;

[0009] L3 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy or C 1-6 hydroxyalkyl;

[0010] Ring A is a monocyclic or bicyclic ring, specifically selected from C 3-10 cycloalkyl, 3-10 membered heterocyclic group, C 6-14 Aryl, 5-10 membered heteroaryl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl, or 5-6 membered heterocyclyl and aryl;

[0011] R 1 Each independently selected from halogen, hydroxyl, amino, amide, C 1-3 Alkylamino, di(C 1-3 alkyl)amino, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group;

[0012] Or, any two R 1 Connect to form a 3-8 membered heterocyclic group or C 3-8 Cycloalkyl;

[0013] R 2 Selected from amino, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl, -OR 2.1 or -NHR 2.1 ; the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl, optionally further substituted by one or more R 2.1 replace;

[0014] R2.1 Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl and 3-6 membered heterocyclic groups, optionally substituted by one or more R 2.1.1 replace;

[0015] R 2.1.1 Selected from halogen, amino, cyano, hydroxyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group;

[0016] R 3 Each independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl or C 1-6 haloalkoxy;

[0017] R 4 is selected from 5-6 membered heteroaryl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl, 5-6 membered heterocyclylphenyl or 5-6 membered heterocyclyl, the 5-6 membered heteroaryl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl, 5-6 membered heterocyclylphenyl and 5-6 membered heterocyclyl, optionally substituted by one or more R 4.1 replace;

[0018] R 4.1 Selected from halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group;

[0019] x and y are each independently selected from 0, 1, 2, 3 or 4.

[0020] In a preferred embodiment of the present invention, the compound of formula (I) is further represented by formula (IA) or formula (IB):

[0021] In a preferred embodiment of the present invention, the aforementioned L1 is selected from -CH2-; or, the R4 is selected from 5-6 membered heteroaryl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl, 5-6 membered heterocyclylphenyl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl or 6 membered heterocyclyl; preferably, the R 4 is selected from thiazolyl, pyrazolyl, pyridazinyl, imidazolyl, pyridinyl, isoxazolyl, triazolyl, tetrazolyl, oxazolyl, pyrazinyl, pyrimidinyl, oxadiazolyl, pyridonyl, pyridazinonyl, pyrimidonyl, pyrazinonyl, imidazopyridinyl, imidazophenyl, thiazophenyl, oxazolophenyl, pyrimidonphenyl, pyrimidonopyridinyl, pyrazolophenyl, pyrimidonphenyl, pyrazinonphenyl, pyrimidonothiazolyl, pyrimidonopyrazolyl or oxazolopyridinyl; more preferably, said R 4 is selected from thiazolyl, pyridazinyl, thiazolophenyl or pyrazinyl.

[0022] In a certain embodiment of the present invention, L3 as described above is an amino group.

[0023] In a preferred embodiment of the present invention, the R 4 is selected from pyridazinyl.

[0024] In a preferred embodiment of the present invention, the compound of formula (IA) described above is further represented by formula (II):

[0025] In a certain embodiment of the present invention, L2 described in the present invention can also be selected from a bond, -CH2-, and -(CH2)2-.

[0026] In a certain embodiment of the present invention, L3 of the present invention can also be selected from C 2-6 Alkenyl, C 2-6 Alkynyl, -P(=O)R 1.1 R 1.2 、-S(=O)2R 1.1 、-S(=O)(=NR 1.1 )R 1.2 、C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 3-6 Cycloalkyl C 1-3 Alkyl, 3-6 membered heterocyclic group C 1-3 Alkyl, -OR 1.1 ;

[0027] R 1.1 and R 1.2 Each independently selected from halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 1- 6 haloalkyl, C1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group.

[0028] In a preferred embodiment of the present invention, L3 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl, C 1- 4-halogenated alkoxy or C 1-4 Hydroxyalkyl.

[0029] In a preferred embodiment of the present invention, L3 is selected from C 2-4 Alkenyl, C 2-4 Alkynyl, -P(=O)R 1.1 R 1.2 、-S(=O)2R 1.1 、-S(=O)(=NR 1.1 )R 1.2 、C 3-4 Cycloalkyl, 3-4 membered heterocyclic group, C 3-4 Cycloalkyl C 1-3 Alkyl, 3-4 membered heterocyclic C 1-3 Alkyl, -OR 1.1 .

[0030] In a preferred embodiment of the present invention, the R 1.1 and R 1.2 Each independently selected from halogen, hydroxy, cyano, amino, C 1-3 Alkyl, C 1-3 Hydroxyalkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl, C 1-3 Halogenated alkoxy, C 3-4 Cycloalkyl, 3-4 membered heterocyclic group.

[0031] In a preferred embodiment of the present invention, x as described above is 0, 1, 2 or 3; preferably 0, 1 or 2.

[0032] In a preferred embodiment of the present invention, the ring A is selected from 5-6 membered heteroaryl, 5 membered heteroarylphenyl, 5 membered heterocyclylphenyl, 5 membered heterocyclyl and 6 membered heteroaryl, 6 membered heterocyclyl and 5 membered heteroaryl, C 6-10 Aryl, C 3-8 cycloalkyl or 3-8 membered heterocyclic group; preferably, the ring A is selected from pyridazinyl, pyrazolyl, pyridyl, phenyl, pyrrolyl, imidazolyl, triazolyl, tetrazolyl, furyl, thienyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyranyl, pyrrolidinyl, pyridonyl, morpholinyl, piperidinyl, oxirane, aziridine, oxetanyl, azetidinyl, piperazinyl, pyrrolophenyl, pyrrolidinylphenyl, diazaspiro[3.3]heptanyl or oxazaspiro[3.3]heptanyl.

[0033] In a certain embodiment of the present invention, the ring A of the present invention is selected from tetrahydrofuranyl, 2-azaspiro[3.3]heptane, pyrrolidinone, azaspiro[2.5]octanyl, cyclohexenyl,

[0034] In a preferred embodiment of the present invention, the R 1 Each independently selected from halogen, hydroxyl, amino, amide, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-4 Cycloalkyl or 3-4 membered heterocyclic group; preferably, the R 1 Each is independently selected from fluorine, chlorine, hydroxy, amino, amido, cyano, methyl, ethyl, propyl, methoxy, ethoxy, propoxy, trifluoromethyl, difluoromethyl, trifluoroethyl, hydroxymethyl, hydroxyethyl, cyclopropyl, cyclobutyl, oxirane, aziridine, oxetanyl or azetidinyl.

[0035] In a preferred embodiment of the present invention, the compound of formula (IB) described above is further represented by formula (III):

[0036] In a preferred embodiment of the present invention, L3 described above is selected from a bond, methylene, ethylene, propylene, halogen-substituted methylene, halogen-substituted ethylene, halogen-substituted propylene, hydroxy-substituted methylene, hydroxy-substituted ethylene or hydroxy-substituted propylene.

[0037] In a preferred embodiment of the present invention, L3 as described above is selected from a bond, -CH2-, -CH2CH2-, -CH(CH3)CH2- or -CH2-CH(F)-CH2-.

[0038] In a preferred embodiment of the present invention, L2 as described above is selected from a bond, -O-, -S- or -NH-.

[0039] In a certain embodiment of the present invention, L2 described in the present invention is selected from -NH-.

[0040] In a preferred embodiment of the present invention, the R 1 Selected from C 1-3 Alkyl, C1-3 Alkoxy, C 1-3 Hydroxyalkyl, C 1- 3 haloalkyl, C 1-3 Alkyl-substituted amino or -SF5.

[0041] In a preferred embodiment of the present invention, the R 1 Selected from methyl, ethyl, methoxy, ethoxy, hydroxymethyl, hydroxyethyl, -CHF2, -CF3, -NHCH3 or -SF5.

[0042] In a preferred embodiment of the present invention, the R 1 Each independently selected from chlorine, fluorine, methoxy, methyl, -CH2F, cyclopropyl, -CF3, or hydroxyl groups.

[0043] In a preferred embodiment of the present invention, the above L2 is a bond, -CH2-, -CF2-, -C(O)-, -NH-, -O- or -S-, wherein the -CH2- and -NH- are optionally replaced by one or two R L2 Replacement, R L2 Each independently selected from halogen, cyano, C 1-3 Alkyl, C 1- 3 alkoxy, C 1-3 Halogenated alkyl, C 1-3 Halogenated alkoxy, C 3-4 In a preferred embodiment of the present invention, the R L2 Each is independently selected from fluoro, chloro, methyl, ethyl, propyl, methoxy, ethoxy, trifluoromethyl, difluoromethyl, trifluoroethyl, cyclopropyl, cyclobutyl, aziridine, oxirane, azetidinyl or oxetanyl.

[0044] In a preferred embodiment of the present invention, L2 as described above is selected from -NH-, -CH2-, -C(O)NH-, -NHC(O)-, -CF2-, -O- or -S-.

[0045] In a preferred embodiment of the present invention, L2 as described above is a bond, -O-, -S-, -NH- or -CH2-.

[0046] In a preferred embodiment of the present invention, the R 2 Selected from amino, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, -OR 2.1 or -NHR2.1 Preferably, the R 2 Selected from amino, methyl, ethyl, propyl, n-butyl, isobutyl, cyclopropyl, piperidinyl, morpholinyl, piperazinyl, -OR 2.1 or -NHR 2.1 ;

[0047] R 2.1 Selected from C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-4 Cycloalkyl, 3-4 membered heterocyclic group, the C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkyl and 3-6 membered heterocyclic groups, optionally substituted by one or more R 2.1.1 substituted; preferably, R 2.1 is selected from methyl, ethyl, propyl, n-butyl, isobutyl, methoxy, ethoxy, propoxy, cyclopropyl, cyclobutyl, cyclopentyl, oxirane, aziridine, oxetanyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, trifluoromethyl, difluoromethyl or difluoroethyl;

[0048] The R 2.1.1 Selected from halogen, amino, cyano, hydroxyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; preferably, the R 2.1.1 is selected from fluorine, chlorine, methyl, ethyl, propyl, n-butyl, isobutyl, methoxy, ethoxy, propoxy, cyclopropyl, cyclobutyl, cyclopentyl, oxirane, aziridine, oxetanyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, trifluoromethyl, difluoromethyl or difluoroethyl;

[0049] Or, R 3 Selected from hydrogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl or C 1-4 preferably, R 3 is selected from hydrogen, methyl, ethyl, propyl, methoxy, ethoxy, propoxy, trifluoromethyl or difluoromethyl.

[0050] In a preferred embodiment of the present invention, the R 3 It is a methyl group.

[0051] In a preferred embodiment of the present invention, the R 2 Selected from

[0052] In a preferred embodiment of the present invention, the R 2 for

[0053] In a preferred embodiment of the present invention, the R 4.1 Selected from fluorine, chlorine, hydroxyl, cyano, amino, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl, C 1-4 Hydroxyalkyl, C 1-4 Halogenated alkoxy, C 3-4 Cycloalkyl or 3-4 membered heterocyclic group; preferably, the present invention as described above R 4.1 is selected from fluorine, chlorine, hydroxy, cyano, amino, methyl, ethyl, propyl, methoxy, ethoxy, propoxy, trifluoromethyl, difluoromethyl, hydroxymethyl, cyclopropyl or cyclobutyl.

[0054] In a preferred embodiment of the present invention, the present invention as described above Selected from

[0055] In a preferred embodiment of the present invention, the present invention as described above Selected from

[0056] In a certain embodiment of the present invention, the present invention Selected from

[0057] In a preferred embodiment of the present invention, the present invention as described above Selected from methyl,

[0058] In a preferred embodiment of the present invention, the and-L3-R 1 Each independently selected from methyl, methoxy, ethynyl, vinyl, -CF3, -OCF3,

[0059] In a preferred embodiment of the present invention, the compound described above is selected from the following compounds in Table 1:

[0060] Table 1

[0061] The present invention provides a pharmaceutical composition comprising:

[0062] (1) the compound represented by formula I as described above or a pharmaceutically acceptable salt thereof, and

[0063] (2) Pharmaceutically acceptable excipients.

[0064] The present invention provides a use of a compound represented by formula I as described above, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, in the preparation of a medicament, wherein the medicament can be used to prevent and / or treat pain; preferably, the pain is neuropathic pain.

[0065] The present invention provides a use of the compound represented by formula I or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above, in the preparation of a drug for a disease mediated by AT2R.

[0066] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0067] Explanation of terms

[0068] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0069] The term "oxo" refers to =0, an oxygen atom replacing two hydrogens on the same carbon atom, ie, a carbonyl replacing a methylene group.

[0070] The term "thio" refers to =S, where a sulfur atom replaces two hydrogen atoms on the same carbon atom.

[0071] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 6 carbon atoms, and more preferably an alkyl group containing 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, and various branched isomers thereof. Methyl, ethyl, isopropyl, isobutyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl, and hydroxy-substituted alkyl are preferred in the present invention.

[0072] The term "cycloalkyl" refers to a saturated or partially unsaturated cyclic hydrocarbon substituent, including monocyclic alkyl, spirocyclic alkyl and bridged cycloalkyl. The cycloalkyl ring comprises 3 to 10 carbon atoms, preferably 3 to 8 carbon atoms, more preferably 3 to 4 carbon atoms. The limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl etc.

[0073] The cycloalkyl group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0074] The term "heterocyclyl" refers to a saturated or partially unsaturated cyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is a heteroatom selected from nitrogen, oxygen, C(O), and S, but excluding the ring moieties of -OO-, -OS-, or -SS-, and the remaining ring atoms are carbon, including monoheterocyclyl, spiroheterocyclyl, and bridged heterocyclyl. Preferably, it contains 3 to 10 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 8 ring atoms; and most preferably, it contains 3 to 6 ring atoms. In the present invention, "membered" refers to the number of ring atoms, for example, a 3-6 membered heterocyclyl means a heterocyclyl containing 3-6 ring atoms. Non-limiting examples of heterocyclic groups include oxetane, thietanyl, azetidinyl, tetrahydropyranyl, azepanyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, and the like, preferably oxetane, thietanyl, azetidine, tetrahydrofuranyl, tetrahydropyranyl, 1-aminoylidene-1-oxothiopyran, azepanyl, piperidinyl, piperazinyl, pyridonyl, wait.

[0075] The heterocyclyl group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, thio, carboxyl or carboxylate.

[0076] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (ie, rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. More preferably, phenyl.

[0077] Aryl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, -P(=O)R 5-1 R 5-2 、-NHC(O)R 5-1 、-NHS(O)2R 5-1 , carboxyl or carboxylate groups.

[0078] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably a 5-8 membered monoheteroaryl or an 8-10 membered diheteroaryl group, more preferably a 5-membered monoheteroaryl, a 6-membered monoheteroaryl, an 8-membered diheteroaryl, a 9-membered bicyclic heteroaryl or a 10-membered bicyclic heteroaryl group, for example, an imidazolyl, a furyl, a thienyl, a thiazolyl, a pyrazolyl, a pyrrolyl, a triazolyl, a tetrazolyl, a pyridyl, a pyrimidinyl, a pyrazinyl, a pyridazinyl, a piperazinyl, a thienopyrrolyl, a thiazolopyrrolyl, an oxazolopyrrolyl, a furopyrrolyl, a thienylfuryl, an oxazoloimidazolyl, a thiazolofuryl, a thiadiazolidinidazolyl, an oxazoloimidazolyl, a pyrrolophenyl, a thienothiazolyl, wait.

[0079] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0080] The term "alkoxy" refers to-O-(alkyl) and-O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as described above. The limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0081] "Haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above. Non-limiting examples include: trifluoromethyl, difluoromethyl.

[0082] "Haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

[0083] "Hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above. Non-limiting examples include: -CH2OH, -C(CH3)2(OH).

[0084] "Amide" means where R A and R B Each independently selected from hydrogen or C 1-6 Alkyl; preferably R A and R B Each independently selected from hydrogen or C 1-3 Alkyl; More preferably, R A and R B Each is independently selected from hydrogen, methyl or ethyl.

[0085] Different expressions such as “X is selected from A, B, or C”, “X is selected from A, B and C”, “X is A, B or C”, and “X is A, B and C” all express the same meaning, that is, X can be any one or more of A, B, and C.

[0086] In the present invention Indicates that the key may not exist;

[0087] The substituents of the present invention It indicates the position where the substituent is attached to the substituted site.

[0088] In the present invention, the "-" at the end of a group means that the group is connected to the rest of the molecule through this site. For example, -CH2- represents a methylene group.

[0089] Those skilled in the art will understand that according to the conventions used in the art, the structural formulas used in this application to describe groups Refers to the connecting bond, the corresponding group through Connect with other fragments and groups in the compound. When the group is O, it represents a double bond (such as =O) or a single bond (such as O - ).

[0090] The hydrogen atoms described in the present invention may be replaced by their isotope deuterium. Any hydrogen atom in the example compounds of the present invention may also be replaced by a deuterium atom.

[0091] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0092] "One or more" means 1, 2, 3, 4, 5, 6, 7, 8, 9 or more, preferably 1, 2, 3 or 4.

[0093] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0094] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0095] "Pharmaceutically acceptable" or "pharmaceutically acceptable" means relatively non-toxic, safe, and suitable for use by patients.

[0096] A "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0097] In the present invention, "NA" means "not tested".

[0098] Unless otherwise specified, all technical and scientific terms used herein have the standard meanings in the art to which the claimed subject matter belongs. If there are multiple definitions for a term, the definition herein shall prevail.

[0099] On the basis of conforming to the common sense in this field, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0100] The reagents and raw materials used in the present invention are commercially available or prepared by methods disclosed in the prior art.

[0101] Unless otherwise specified, the chiral carbon atoms in the compounds of the present invention are in the R or S configuration.

[0102] The general preparation method of the compounds of the present invention is as follows:

[0103] Where: X is halogen, M1, M2, L, L1, L2, R 1 、R 2 、R 3 、R 4 The definitions of x and y are the same as those described above in the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0104] FIG1 is the timeline of Section 1.7 of Test Example 5.

[0105] Figure 2 shows the timeline of Section 2.2.7 of Test Case 6. DETAILED DESCRIPTION

[0106] The present invention is further illustrated by way of examples, but the invention is not limited to the scope of these examples. Experimental methods in the following examples, where specific conditions are not specified, were performed according to conventional methods and conditions, or according to the product specifications. Unless otherwise specified, the reagents used in the present invention were commercially available or prepared according to methods disclosed in the prior art.

[0107] Preparation of intermediate A

[0108] Step 1: Preparation of compound A-2

[0109] Under nitrogen protection, N,N-diisopropylethylamine (5.81 g, 44.94 mmol) and 3-(chloromethyl)pyridazine (2.31 g, 17.97 mmol) were added to a solution of compound A-1 (3.0 g, 14.98 mmol) in N,N-dimethylformamide (30 mL) at room temperature. The reaction mixture was heated to 60°C and stirred for 16 hours. After the reaction was completed, the reaction mixture was quenched by adding ice water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-5%) to obtain compound A-2 (2.5 g, yield 57.08%). LCMS (ESI, m / z): 293.0 [M+H] + .

[0110] Step 2: Preparation of Intermediate A

[0111] To a solution of compound A-2 (2.5 g, 8.55 mmol) in dichloromethane (30 mL) was added a 1,4-dioxane solution (8 mL, 4 M) of hydrochloric acid at room temperature. The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated, and the resulting residue was purified to afford the product (S)-3-((2-methylpiperazin-1-yl)methyl)pyridazine hydrochloride (2.4 g of Intermediate A, crude product) as a yellow solid. LCMS (ESI, m / z): 193.0 [M+H] + .

[0112] Example 1 Preparation of Compound 1

[0113] Step 1: Preparation of compound 1-2

[0114] Under nitrogen, potassium carbonate (6 g, 29 mmol) and benzyl bromide (5 g, 29 mmol) were added to a solution of (1R,2R)-2-aminocyclopentanol hydrochloride (2 g, 14.5 mmol) in acetone / water (v:v = 5:1) (55 mL) at room temperature. The reaction was allowed to proceed at 60°C for 16 h, and the desired product was observed in the liquid phase. The reaction solution was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The resulting residue was directly purified by column chromatography (ethyl acetate / petroleum ether = 10%) to afford compound 1-2 (3.8 g, 92%) as a yellow oil. LCMS (ESI, m / z): 282.1 [M+H] + .

[0115] Step 2: Preparation of Compounds 1-3

[0116] Under hydrogen protection, sodium hydride (60% dispersion in mineral oil) (1.1 g, 27 mmol) was added to a solution of compound 1-2 (3.8 g, 13.5 mmol) in tetrahydrofuran (50 mL) at 0°C and allowed to react at room temperature for 1 h. Methyl iodide (2.9 g, 20.25 mmol) was added under ice-cooling and allowed to react at room temperature for 16 h. The desired product was detected in the liquid phase, and the reaction solution was poured into ice water (50 mL). The system was extracted with ethyl acetate (2 x 50 mL), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to dryness. The resulting residue was directly purified by column chromatography (ethyl acetate / petroleum ether = 10%) to afford compound 1-3 (3.8 g, 95%) as a colorless oil. LCMS (ESI, m / z): 296.1 [M+H] + .

[0117] Step 3: Preparation of Compounds 1-4

[0118] Under hydrogen protection, Pd / C (310 mg) was added to a solution of compound 1-3 (3.1 g, 10.5 mmol) in methanol (40 mL) at room temperature. The mixture was allowed to react for 16 h at room temperature. The desired product was found in the liquid phase, which was then filtered and concentrated to dryness. The resulting residue was directly purified by column chromatography (methanol / dichloromethane = 5%) to afford compound 1-4 (700 mg, 58%) as a colorless oil. LCMS (ESI, m / z): 116.2 [M+H] + .

[0119] Step 4: Preparation of Compounds 1-6

[0120] Under nitrogen, to a solution of compound 1-4 (400 mg, 3.48 mmol) in dimethyl sulfoxide (5 mL) were added compound 1-5 (890 mg, 3.48 mmol) and N,N-diisopropylethylamine (1.3 g, 10.44 mmol), respectively. The mixture was heated to 120°C and allowed to react for 16 h. The desired product was detected in the liquid phase, and the reaction solution was poured into water (20 mL). The system was extracted with ethyl acetate (2 x 30 mL). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to dryness. The resulting residue was directly purified by column chromatography (methanol / dichloromethane = 5%) to afford compound 1-6 (700 mg, 58%) as a yellow solid. LCMS (ESI, m / z): 351.0 [M+H] + .

[0121] Step 5: Preparation of Compounds 1-7

[0122] Under nitrogen, to a solution of compound 1-6 (285 mg, 0.82 mmol) in anhydrous 1,4-dioxane (5 mL) were added intermediate A hydrochloride (187.4 mg, 0.82 mmol), cesium carbonate (932 mg, 2.87 mmol), and (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (70 mg, 0.082 mmol). The reaction was heated to 110°C for 16 h. The desired product was detected in the liquid. The reaction solution was poured into water (20 mL). The system was extracted with ethyl acetate (2 x 30 mL). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to dryness. The resulting residue was directly purified by column chromatography (methanol / dichloromethane = 5%) to obtain compound 1-7 (200 mg, 53%) as a yellow oil. LCMS (ESI, m / z): 463.4 [M+H] + .

[0123] Step 6: Preparation of Compound 1

[0124] Compound 1-7 (100 mg, 0.21 mmol) and toluene (2 mL) were added to a 10 mL sealed tube. Tri-n-butyltin chloride (352 mg, 1.08 mmol) and sodium azide (71 mg, 1.08 mmol) were then added, respectively. The mixture was reacted at 140°C for 48 hours with liquid chromatography-mass spectrometry. After completion of the reaction, the residue was concentrated under reduced pressure. The residue was directly purified by column chromatography (methanol / dichloromethane = 5%) to yield 120 mg of a crude yellow solid. The crude product was purified by preparative high-performance liquid chromatography (HPLC) under the following conditions: (Chromatographic column: Sunfire C18 5m, 19mm*250mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 ml / min; elution gradient: 30% B to 49% B over 17 minutes; detection wavelength: UV 254nm / 214nm; retention time (min): 7.6-9.4) to obtain compound 1 (10.45 mg, 9.8%). LCMS (ESI, m / z): 506.4 [M+H] + .

[0125] 1H NMR(400MHz,DMSO-d6)δ9.14-9.09(m,1H),7.72-7.62(m,2H),6.42(s,1H),6.39-6.25(m,2 H),4.15(d,J=14.4Hz,1H),3.68-3.60(m,2H),3.50-3.47(m,1H),3.23(s,3H),2.76-2.64( m,3H),2.60-2.53(m,2H),2.47-2.33(m,4H),2.11-2.02(m,1H),1.92-1.82(m,1H),1.77-1 .68(m,1H),1.66-1.56(m,3H),1.38-1.28(m,1H),0.95(d,J=6.0Hz,3H),0.93-0.88(m,6H).

[0126] Example 2 Preparation of Compound 2

[0127] Step 1: Preparation of compound 2-2

[0128] Under nitrogen protection, sodium hydroxide (281.0 mg, 7.03 mmol) and 2-methoxyethane-1-amine (294.0 mg, 3.51 mmol) were added to a solution of compound 1-5 (600.0 mg, 2.34 mmol) in acetonitrile and water (10 mL / 2 mL) at room temperature. The reaction mixture was heated to 80°C and stirred for 16 hours. After the reaction was completed, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-20%) to obtain the product compound 2-2 (200.0 mg, yield 27.4%) as a yellow oil. LCMS (ESI, m / z): 311.0 [M+H] + .

[0129] Step 2: Preparation of compound 2-3

[0130] To a solution of compound 2-2 (200.0 mg, 0.64 mmol) in 1,4-dioxane (5 mL) and water (1 mL) at room temperature were added intermediate A hydrochloride (147.0 mg, 0.64 mmol), methanesulfonic acid [2-(dicyclohexylphosphino)-2'-(N,N-dimethylamino)-1,1'-biphenyl] (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (49 mg, 0.064 mmol), and cesium carbonate (628.0 mg, 1.93 mmol). The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to obtain the product compound 2-3 (50.0 mg, yield 18.41%) as a yellow solid. LCMS (ESI, m / z): 423.1 [M+H] + .

[0131] Step 3: Preparation of Compound 2

[0132] Under nitrogen protection, sodium azide (38.46 mg, 0.591 mmol) and tri-n-butyltin azide (192.57 mg, 0.591 mmol) were added to a toluene (2 mL) solution of compound 2-3 (50.0 mg, 0.118 mmol) in a microwave tube. The reaction mixture was heated to 140° C. and stirred for 16 hours. After the reaction, the reaction solution was filtered through potassium fluoride solid wash, and the solid was washed with ethyl acetate. The filtrate was concentrated, and the resulting residue was directly purified by column chromatography with methanol / dichloromethane (0-10%) to obtain a crude yellow solid. The crude product was purified by preparative HPLC under the following conditions: (Chromatographic column specifications: Sunfire C18 10 μm, 19 mm x 250 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 ml / min; elution gradient: 27-37% B over 18 minutes; detection wavelength: UV 254 nm / 214 nm; retention time (min): 6.8-8.1) to obtain compound 2 (8.58 mg, yield 15.57%). LCMS (ESI, m / z): 466.2 [M+H] + .

[0133] 1H NMR(400MHz,CD3OD)δ9.09(d,J=4.4Hz,1H),7.86(d,J=8.4Hz,1H),7.76-7.66(m ,1H),6.43(s,2H),4.30(d,J=14.4Hz,1H),3.75(d,J=14.4Hz,1H),3.63-3.52(m, 2H),3.34(s,5H),2.97-2.84(m,3H),2.79-2.60(m,3H),2.58-2.48(m,1H),2.46( d,J=7.2Hz,2H),1.99-1.79(m,1H),1.08(d,J=5.6Hz,3H),0.94(d,J=6.4Hz,6H).

[0134] Example 3 Preparation of Compound 3

[0135] Step 1: Preparation of compound 3-1

[0136] Under nitrogen protection, N,N-diisopropylethylamine (3.03 g, 23.43 mmol) and intermediate A (1.5 g, 7.81 mmol) were added to a solution of compound 1-5 (2.0 g, 7.81 mmol) in dimethyl sulfoxide (20 mL) at room temperature. The reaction mixture was heated to 120°C and stirred for 16 hours. After the reaction was completed, the reaction mixture was quenched by adding ice water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-20%) to obtain the product compound 3-1 (2.3 g, yield 68.76%) as a yellow oil. LCMS (ESI, m / z): 428.1 [M+H] + .

[0137] Step 2: Preparation of compound 3-2

[0138] To a solution of compound 3-1 (300.0 mg, 0.7 mmol) in 1,4-dioxane (5 mL) were added 1-methyl-1H-pyrazol-4-amine (102.0 mg, 1.05 mmol), methanesulfonic acid [2-(dicyclohexylphosphino)-2'-(N,N-dimethylamino)-1,1'-biphenyl] (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (54 mg, 0.07 mmol), and cesium carbonate (685 mg, 2.1 mmol) at room temperature. The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to obtain the product compound 3-2 (83.0 mg, yield 26.66%) as a yellow solid. LCMS (ESI, m / z): 445.1 [M+H] + .

[0139] Step 3: Preparation of Compound 3

[0140] Under nitrogen protection, sodium azide (61 mg, 0.933 mmol) and tri-n-butyltin azide (303.8 mg, 0.933 mmol) were added to a toluene (2 mL) solution of compound 3-2 (83.0 mg, 0.187 mmol) in a microwave tube. The reaction mixture was heated to 140° C. and stirred for 16 hours. After the reaction, the reaction solution was filtered through potassium fluoride solid wash, and the solid was washed with ethyl acetate. The filtrate was concentrated, and the resulting residue was directly purified by column chromatography with methanol / dichloromethane (0-10%) to obtain a crude yellow solid. The crude product was purified by preparative HPLC under the following conditions: (Chromatographic column specifications: Sunfire C18 10 μm, 19 mm x 250 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 ml / min; elution gradient: 20-30% B over 17 minutes; detection wavelength: UV 254 nm / 214 nm; retention time (min): 7.5-9.6) to obtain compound 3 (14.89 mg, yield 16.36%). LCMS (ESI, m / z): 488.1 [M+H] + .

[0141] 1H NMR (400MHz, CD3OD) δ9.17-9.01(m,1H),7.92-7.80(m,1H),7.78-7.66(m,1H),7.55 (s,1H),7.37(s,1H),6.52(d,J=11.2Hz,2H),4.34(d,J=14.4Hz,1H),3.87(s,3H),3 .80(d,J=14.4Hz,1H),3.03-2.89(m,3H),2.81-2.67(m,3H),2.62-2.52(m,1H),2.3 8(d,J=7.2Hz,2H),1.93-1.74(m,1H),1.11(d,J=6.0Hz,3H),0.89(d,J=6.4Hz,6H).

[0142] Example 4 Preparation of Compound 4

[0143] Step 1: Preparation of compound 4-1

[0144] Under nitrogen protection, N,N-diisopropylethylamine (756.96 mg, 5.86 mmol) and cyclopropaneamine (222.9 mg, 3.90 mmol) were added to a solution of compound 1-5 (500.0 mg, 1.95 mmol) in dimethyl sulfoxide (5 mL) at room temperature. The reaction mixture was heated to 120°C and stirred for 16 hours. After the reaction was completed, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-20%) to obtain the product compound 4-1 (160.0 mg, yield 27.95%) as a yellow oil. LCMS (ESI, m / z): 333.9 [M+CH3CN] + .

[0145] Step 2: Preparation of compound 4-2

[0146] To a solution of compound 4-1 (160.0 mg, 0.54 mmol) in 1,4-dioxane (5 mL) and water (1 mL) at room temperature were added intermediate A hydrochloride (124.8 mg, 0.54 mmol), [2-(dicyclohexylphosphino)-2'-(N,N-dimethylamino)-1,1'-biphenyl](2'-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (42 mg, 0.054 mmol), and cesium carbonate (533.4 mg, 1.64 mmol). The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to obtain the product compound 4-2 (50.0 mg, yield 22.65%) as a yellow solid. LCMS (ESI, m / z): 405.2 [M+H] + .

[0147] Step 3: Preparation of compound 4

[0148] Under nitrogen protection, sodium azide (40.17 mg, 0.618 mmol) and tri-n-butyltin azide (201.15 mg, 0.618 mmol) were added to a toluene (2 mL) solution of compound 4-2 (50.0 mg, 0.123 mmol) in a microwave tube. The reaction mixture was heated to 140° C. and stirred for 16 hours. After the reaction, the reaction solution was filtered through potassium fluoride solid wash, and the solid was washed with ethyl acetate. The filtrate was concentrated, and the resulting residue was directly purified by column chromatography with methanol / dichloromethane (0-10%) to obtain a crude yellow solid. The crude product was purified by preparative HPLC under the following conditions: (Chromatographic column specifications: Sunfire C18 10 μm, 19 mm x 250 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 ml / min; elution gradient: 28-38% B over 17 minutes; detection wavelength: UV 254 nm / 214 nm; retention time (min): 8.5-10) to obtain compound 4 (2.83 mg, yield 5.12%). LCMS (ESI, m / z): 448.1 [M+H] + .

[0149] 1H NMR (400MHz, CD3OD) δ9.09 (dd, J=4.9, 1.6Hz, 1H), 7.87 (dd, J=8.5, 1.6Hz, 1H), 7.72 (dd, J=8. 5,4.9Hz,1H),6.83(s,1H),6.47(s,1H),4.31(d,J=14.5Hz,1H),3.76(d,J=14.4Hz,1H),2.93( d,J=9.4Hz,3H),2.83–2.64(m,3H),2.56(dd,J=12.2,6.2Hz,1H),2.51–2.40(m,3H),2.05–1. 81(m,1H),1.09(d,J=5.8Hz,3H),0.95(d,J=6.6Hz,6H),0.83–0.72(m,2H),0.53–0.42(m,2H).

[0150] Example 5 Preparation of Compound 5

[0151] Step 1: Preparation of compound 5-1

[0152] Under nitrogen protection, to a solution of (S)-2-bromo-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile (250.0 mg, 0.58 mmol, 1.0 equiv) in dioxane (5 mL) was added hydroxymethyltributylstannane (282.0 mg, 0.87 mmol, 1.5 equiv) and tetrakis(triphenylphosphine)palladium (67.0 mg, 0.058 mmol, 0.1 equiv) in a microwave tube. The reaction mixture was heated to 100°C and stirred for 16 hours. After completion of the reaction, the mixture was quenched with water and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford the yellow compound 5-1 (130 mg, 59.1% yield). LCMS: (ESI, m / z): 380.2 [M+H] + .

[0153] Step 2: Preparation of compound 5-2

[0154] To a solution of (S)-2-(hydroxymethyl)-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile (130.0 mg, 0.34 mmol, 1.0 equiv) in dichloromethane (5 mL) was added triethylamine (103.0 mg, 1.02 mmol, 3.0 equiv) and methanesulfonyl chloride (78.2 mg, 0.68 mmol, 2.0 equiv) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was quenched by addition of ice water (10 mL) and extracted with dichloromethane (10 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to afford the yellow compound 5-2 (160 mg, crude product). LCMS: (ESI, m / z): 458.2 [M+H] + .

[0155] Step 3: Preparation of compound 5-3

[0156] To a solution of (S)-2-cyano-5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzyl methanesulfonate (160.0 mg, 0.35 mmol, 510 equiv) in N,N-dimethylformamide (5 mL) was added imidazole (119.0 mg, 1.75 mmol, 5.0 equiv) and potassium carbonate (150.0 mg, 1.05 mmol, 3.0 equiv) and allowed to react at room temperature for 16 hours. The desired product was found in the liquid. After completion of the reaction, the mixture was quenched with water and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford compound 5-3 (120 mg, 80.0% yield) as a yellow oil. LCMS: (ESI, m / z): 430.0 [M+H] + .

[0157] Step 4: Preparation of compound 5

[0158] Under nitrogen protection, sodium azide (91.0 mg, 1.4 mmol, 5 equiv) and tri-n-butyltin azide (456.0 mg, 1.4 mmol, 5 equiv) were added to a solution of ((S)-2-((1H-imidazol-1-yl)methyl)-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile (120.0 mg, 0.28 mmol, 1.0 equiv) in toluene (4 mL) in a microwave tube. The reaction mixture was heated to 140°C and stirred for 2 days. After the reaction, the reaction solution was washed with potassium fluoride solid and filtered. The solid was washed with ethyl acetate, and the filtrate was concentrated. The resulting residue was directly purified by column chromatography with methanol / dichloromethane (0-10%) to obtain a crude yellow solid. The crude product was purified by preparative HPLC under the following purification conditions: (Chromatographic column specifications: Sunfire C18 10 μm, 19 mm x 250 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 ml / min; elution gradient: 5-15% B over 16 minutes; detection wavelength: UV 254 nm / 214 nm; retention time (min): 9.2-10.5) to obtain compound 5 (30.17 mg, yield 22.8%). LCMS: (ESI, m / z): 473.2 [M+H] + .

[0159] 1 H NMR(400MHz,DMSO-d6)δ9.11-9.10(m,1H),8.13(s,0.62HCOOH),7.65-7.64(m,2H ),7.40(s,1H),6.99(s,1H),6.91(s,1H),6.86(s,1H),6.79(s,1H),5.00(s,2H),4 .06(d,J=14.0Hz,1H),3.59(d,J=14.0Hz,1H),2.70-2.65(m,3H),2.48-2.44(m,4H ),2.36-2.29(m,1H),2.12-2.05(m,1H),1.86-1.79(m,1H),0.85(d,J=6.4Hz,9H).

[0160] Example 6 Preparation of Compound 6

[0161] Compounds Replacement compounds Referring to the synthesis method of compound 5, compound 6 (11.48 mg, yield 13.14%) was obtained. LCMS: (ESI, m / z): 512.4 [M+H] + .

[0162] 1 H NMR(400MHz,CD3OD)δ9.09(d,J=3.6Hz,1H),7.82(d,J=8.4Hz,1H),7.73-7.69(m,1 H),7.05(s,1H),6.99(s,1H),4.26(d,J=14.4Hz,1H),3.77(d,J=14.4Hz,1H),3.52( s,2H),2.86-2.83(m,3H),2.71-2.61(m,4H),2.57-2.50(m,5H),2.37-2.31(m,1H), 2.12-2.02(m,2H),1.95-1.88(m,1H),0.98(d,J=6.0Hz,3H),0.93(d,J=6.4Hz,6H).

[0163] Example 7 Preparation of Compound 7

[0164] Step 1: Preparation of compound 7-1

[0165] To a solution of (S)-2-(hydroxymethyl)-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile (200.0 mg, 0.528 mmol, 1.0 equiv) in dichloromethane was added Dess-Martin periodinane (336.0 mg, 0.792 mmol, 1.5 equiv) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford (S)-2-formyl-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile compound 7-1 (120.0 mg, yield 60.28%) as a yellow solid. LCMS: (ESI, m / z): 378.4 [M+H] + .

[0166] Step 2: Preparation of compound 7-2

[0167] To a solution of (S)-2-formyl-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile (120.0 mg, 0.318 mmol, 1.0 equiv) in dichloromethane was added diethylaminosulfur trifluoride (128.0 mg, 0.796 mmol, 2.5 equiv) at 0°C. The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford (S)-2-(difluoromethyl)-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile compound 7-2 (68.0 mg, 53.46% yield) as a yellow solid. LCMS: (ESI, m / z): 400.2 [M+H] + .

[0168] Step 3: Preparation of compound 7

[0169] To a solution of (S)-2-(difluoromethyl)-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile (76.0 mg, 0.190 mmol, 1.0 equiv) in toluene (4 mL) was added sodium azide (62.0 mg, 0.952 mmol, 5 equiv) and tri-n-butyltin azide (309.0 mg, 0.952 mmol, 5 equiv) in a microwave tube under nitrogen. The reaction mixture was heated to 140° C. and stirred for 3 days. After the reaction, the reaction solution was filtered through potassium fluoride solid wash, and the solid was washed with ethyl acetate. The filtrate was concentrated, and the resulting residue was directly purified by column chromatography with methanol / dichloromethane (0-10%) to obtain a crude yellow solid. The crude product was purified by preparative HPLC under the following conditions: (Chromatographic column specifications: Sunfire C18 10 μm, 19 mm x 250 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 ml / min; elution gradient: 23-33% B over 16 minutes; detection wavelength: UV 254 nm / 214 nm; retention time (min): 8.9-10.6) to obtain compound 7 (42.63 mg, yield 50.76%). LCMS: (ESI, m / z): 443.5 [M+H] + .

[0170] 1H NMR (400MHz, CD3OD) δ9.13-9.11(m,1H),7.83-7.80(m,1H),7.74-7.71(m,1H),7.28( s,1H),7.18(s,1H),6.47(t,J=55.2Hz,1H),4.39(d,J=14.4Hz,1H),3.94(d,J=14.4H z,1H),2.97-2.92(m,3H),2.86-2.81(m,1H),2.73-2.68(m,2H),2.61(d,J=7.2Hz,2H ),2.55-2.50(m,1H),1.98-1.89(m,1H),1.03(d,J=5.6Hz,3H),0.95(d,J=6.4Hz,6H).

[0171] Example 8 Preparation of Compound 8

[0172] Compounds Replacement compounds Compound 8 (19.44 mg, yield 30.5%) was obtained by referring to the synthesis method of compound 5. LCMS: (ESI, m / z): 487.4 [M+H] + .

[0173] 1 H NMR (400MHz, CD3OD) δ9.08 (d, J=4.8Hz, 1H), 8.24 (s, 0.59HCOOH), 7.85-7.78 (m, 2H), 7.71-7.68 (m,1H),7.03-7.01(m,2H),6.81(s,1H),5.07-5.02(mz,2H),4.23(d,J=14.4Hz,1H),3.78(d,J= 14.4Hz,1H),2.93-2.89(m,2H),2.85-2.81(m,1H),2.70-2.67(m,1H),2.63-2.58(m,1H),2.55- 2.49(m,3H),2.38-2.33(m,1H),2.19(s,2H),2.01(s,1H),1.95-1.85(m,1H),0.96-0.91(m,9H).

[0174] Example 9 Preparation of Compound 9

[0175] Compounds Replacement compounds Referring to the synthesis method of compound 5, (S)-3-((4-(5-isobutyl-3-(4-methyl-1H-pyrazol-1-yl)methyl)-2-(2H-tetrazol-5-yl)phenyl)-2-methylpiperazin-1-yl)methyl ester)pyridazine (compound 9, 22.24 mg, yield 22.53%) was obtained. LCMS: (ESI, m / z): 487.5 [M+H] + .

[0176] 1 H NMR(400MHz,MeOD)δ9.11(d,J=4.4Hz,1H),7.81(d,J=8.4Hz,1H),7.73-7.70(m,1H),7.20(s, 1H),6.99(d,J=5.6Hz,2H),6.73(s,1H),5.08(s,2H),4.33(d,J=14.4Hz,1H),3.85(d,J=14.4 Hz,1H),2.90-2.84(m,3H),2.79-2.73(m,1H),2.66-2.58(mz,2H),2.48(d,J=7.2Hz,2H),2.4 4-2.39(m,1H),1.99(s,3H),1.89-1.82(m,1H),1.00(d,J=5.2Hz,3H),0.89(d,J=6.4Hz,6H).

[0177] Example 10 Preparation of Compound 10

[0178] Compounds Replacement compounds Referring to the preparation method of compound 3, 5-isobutyl-N-(2-methoxycyclohexyl)-3-((S)-3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (Compound 10, 25.88 mg, 8%) was obtained. LCMS: (ESI, m / z): 520 [M+H] + .

[0179] 1H NMR(400MHz,DMSO-d6)δ9.121–9.105(m,1H),7.694–7.633(m,2H),6.449–6.356(m ,2H),6.258(d,J=12.4Hz,1H),4.168–4.133(m,1H),3.640–3.604(m,1H),3.205–3. 139(m,3H),2.742–2.6148(m,3H),2.556–2.526(m,1H),2.450–2.328(m,5H),1.94 6–1.844(m,3H),1.588–1.158(m,8H),0.957–0.922(m,1H),0.889(d,J=6.4Hz,6H).

[0180] Example 11 Preparation of Compound 11

[0181] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-3-((4-(5-isobutyl-3-(1-methyl-1H-pyrazol-4-yl)-2-(2H-tetrazol-5-yl)phenyl)-2-methylpiperazin-1-yl)methyl)pyridazine (Compound 11, 4.69 mg, 9%) was obtained. LCMS: (ESI, m / z): 473.2 [M+H] + .

[0182] 1 H NMR(400MHz,DMSO-d6)δ9.11-9.09(m,1H),7.67-7.62(m,2H),7.27(s,1H),7.1 0(s,1H),6.95(s,1H),6.88(s,1H),4.00(d,J=14.0Hz,1H),3.70(s,3H),3.58( d,J=14.0Hz,1H),2.87-2.68(m,4H),2.58-2.54(m,2H),2.45-2.41(m,1H),2.3 0-2.22(m,1H),2.00-1.88(m,2H),0.92(d,J=6.4Hz,6H),0.83(d,J=6.0Hz,3H).

[0183] Example 12 Preparation of Compound 12

[0184] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-3-((4-(2',6'-difluoro-5-isobutyl-2-(2H-tetrazol-5-yl)-[1,1'-biphenyl]-3-yl)-2-methylpiperazin-1-yl)methyl)pyridazine (Compound 12, 10.07 mg, 4.8%) was obtained. LCMS: (ESI, m / z): 505.2 [M+H] + .

[0185] 1 H NMR(400MHz,DMSO-d6)δ9.11-9.10(M,1H),8.18(s,1H),7.68-7.62(m,2H),7.33- 7.26(m,1H),7.09-7.04(m,1H),6.97-6.86(m,3H),4.05(d,J=14.0Hz,1H),3.58(d ,J=14.4Hz,1H),2.79-2.67(m,3H),2.54(s,3H),2.46(s,1H),2.35-2.30(m,1H),2 .12-2.07(m,1H),1.94-1.86(m,1H),0.90(d,J=6.4Hz,6H),0.86(d,J=6.0Hz,3H).

[0186] Example 13 Preparation of Compound 13

[0187] Step 1: Preparation of compound 13-1

[0188] To a solution of 2-bromo-6-fluoro-4-isobutylbenzonitrile (180.0 mg, 0.702 mmol, 1.0 equiv) in N,N-dimethylformamide (5 mL) was added cesium carbonate (687 mg, 2.11 mmol, 3.0 equiv) and 1-methyl-1H-pyrazol-4-ol (82.7 mg, 0.843 mmol, 1.2 equiv) at room temperature under nitrogen. The reaction mixture was heated to 50°C and stirred for 3 hours. After completion of the reaction, the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-20%) to afford the product (13-1, 210.0 mg, 89.40% yield) as a yellow oil. LCMS: (ESI, m / z): 334.0 [M+H]. + .

[0189] Step 2: Preparation of compound 13-2

[0190] To a solution of compound 13-1 in 1,4-dioxane (5 mL) were added tert-butyl (S)-2-methylpiperazine-1-carboxylate (151.0 mg, 0.754 mmol, 1.2 equiv), (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (52.6 mg, 0.063 mmol, 0.1 equiv), and cesium carbonate (614.2 mg, 1.89 mmol, 3.0 equiv) at room temperature. The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to obtain the product (13-2, 150.0 mg, 52.63% yield) as a yellow solid. LCMS: (ESI, m / z): 454.3 [M+H] + .

[0191] Step 3: Preparation of compound 13-3

[0192] To a solution of compound 13-2 (150 mg, 0.33 mmol, 1.0 equiv) in dichloromethane (3 mL) was added a 1,4-dioxane solution of 4 M hydrochloric acid (2 mL) and allowed to react at room temperature for 2 hours. The desired product was found in the liquid phase. The solution was concentrated under reduced pressure to afford a white solid (13-3, 170.0 mg, crude). LCMS: (ESI, m / z): 354.5 [M+H] + .

[0193] Step 4: Preparation of compound 13-4

[0194] To a solution of compound 13-3 (170.0 mg, 0.436 mmol, 1.0 equiv) in N,N-dimethylformamide (3 mL) at room temperature were added 3-(chloromethyl)pyridazine (67.3 mg, 0.523 mmol, 1.2 equiv) and N,N-diisopropylethylamine (169.0 mg, 1.31 mmol, 3.0 equiv). The reaction mixture was heated to 50°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford the product (13-4, 130.0 mg, 66.92% yield) as a yellow oil. LCMS: (ESI, m / z): 446.7 [M+H] + .

[0195] Step 5: Preparation of compound 13

[0196] Under nitrogen protection, sodium azide (94.8 mg, 1.46 mmol, 5.0 equiv) and tri-n-butyltin azide (474.8 mg, 1.46 mmol, 5.0 equiv) were added to a toluene (3 mL) solution of compound 13-4 (130.0 mg, 0.291 mmol, 1.0 equiv) in a microwave tube. The reaction mixture was heated to 140° C. and stirred for 2 days. After the reaction, the reaction solution was filtered through potassium fluoride solid wash, and the solid was washed with ethyl acetate. The filtrate was concentrated, and the resulting residue was directly purified by column chromatography with methanol / dichloromethane (0-10%) to obtain a crude yellow solid. The crude product was purified by preparative HPLC under the following conditions: (Chromatographic column specifications: Sunfire C18 10 μm, 19 mm x 250 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 20 ml / min; elution gradient: 16-26% B over 16 minutes; detection wavelength: UV 254 nm / 214 nm; retention time (min): 9.2–10.6) to obtain a white compound (13, 31.87 mg, yield 22.36%). LCMS: (ESI, m / z): 489.4 [M+H] + .

[0197] 1H NMR(400MHz,CD3OD)δ9.11-9.09(m,1H),8.14(s,0.12HCOOH),7.84-7.81(m,1H),7. 73-7.70(m,1H),7.45(s,1H),7.23(s,1H),6.74(s,1H),6.57(s,1H),4.30(d,J=14.4 Hz,1H),3.85-3.81(m,4H),2.94-2.84(m,3H),2.78-2.72(m,1H),2.69-2.58(m,2H) ,2.46-2.40(m,3H),1.87-1.77(m,1H),1.02(d,J=6.0Hz,3H),0.89(d,J=6.4Hz,6H).

[0198] Example 14 Preparation of Compound 14

[0199] Compounds Replacement compounds Referring to the preparation method of compound 13, (S)-3-((4-(5-isobutyl-3-(pyridin-3-yloxy)-2-(2H-tetrazol-5-yl)phenyl)-2-methylpiperazin-1-yl)methyl)pyridazine (compound 14, 32.81 mg, yield 20.62%) was obtained. LCMS: (ESI, m / z): 486.2 [M+H] + .

[0200] 1 H NMR (400MHz, CD3OD) δ9.11(d,J=3.2Hz,1H),8.21(d,J=3.2Hz,1H),8.13(d,J=2.0Hz,1H),7 .83(d,J=8.4Hz,1H),7.74-7.70(m,1H),7.36-7.29(m,2H),6.89(s,1H),6.62(s,1H),4.34( d,J=14.8Hz,1H),3.87(d,J=14.0Hz,1H),3.00-2.87(m,3H),2.83-2.76(m,1H),2.73-2.62 (m,2H),2.52-2.47(m,3H),1.92-1.82(m,1H),1.02(d,J=6.0Hz,3H),0.92(d,J=6.4Hz,6H).

[0201] Example 15 Preparation of Compound 15

[0202] Compounds Replacement compounds Referring to the synthesis method of compound 13, (S)-N-(2,2-difluoroethyl)-5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (compound 15, 16.29 mg, yield 8.71%) was obtained. LCMS: (ESI, m / z): 472.4 [M+H] + ,RT(min):1.353.

[0203] 1 H NMR(400MHz,CD3OD)δ9.10-9.09(m,1H),7.88-7.85(m,1H),7.74-7.70(m,1H),6.5 1(d,J=14.4Hz,2H),6.08-5.78(m,1H),4.34(d,J=14.4Hz,1H),3.79(d,J=14.4Hz,1 H),3.65-3.57(m,2H),2.94-2.91(m,3H),2.81-2.66(m,3H),2.60-2.53(m,1H),2.4 7(d,J=7.2Hz,2H),1.95-1.88(m,1H),1.09(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H).

[0204] Example 16 Preparation of Compound 16

[0205] Compounds Replacement compounds Referring to the synthesis method of compound 13, 2-((5-isobutyl-3-((S)-3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)amino)cyclopentan-1-ol was obtained (Compound 16, 36.65 mg, yield 25.74%). LCMS: (ESI, m / z): 492.6 [M+H] + .

[0206] 1H NMR(400MHz,CD3OD)δ9.18(d,J=3.6Hz,1H),7.88-7.86(m,1H),7.79-7.76(m,1H),6.61(s,1H),6.46(s,1H), 4.64(d,J=13.6Hz,1H),4.16(d,J=14.8Hz,1H),4.01-3.97(m,1H),3.67-3.64(m,1H),3.21-3.12(m,2H),3.0 7-3.03(m,3H),2.97-2.92(m,1H),2.88-2.83(m,1H),2.49(d,J=7.2Hz,2H),2.27-2.22(m,1H),1.97-1.87(m ,2H),1.85-1.71(m,2H),1.66-1.60(m,1H),1.52-1.42(m,1H),1.25(d,J=6.0Hz,3H),0.95(d,J=6.4Hz,6H).

[0207] Example 17 Preparation of Compound 17

[0208] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)-3,5-dimethylisoxazol-4-amine (Compound 17, 28.66 mg, yield 23.89%) was obtained. LCMS: (ESI, m / z): 503.4 [M+H] + .

[0209] 1 H NMR (400MHz, CD3OD) δ9.26 (s, 1H), 7.83 (d, J = 2.8Hz, 2H), 6.58 (s, 1H), 6.14 (s, 1H), 3.57-3.40 (m, 4H), 3.21-3.12 (m, 4H), 3.0 4-2.93(m,1H),2.41(d,J=7.2Hz,2H),2.24(s,3H),2.04(s,3H),1.85-1.79(m,1H),1.36-1.29(m,3H),0.89(d,J=6.4Hz,6H).

[0210] Example 18 Preparation of Compound 18

[0211] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)-1-methyl-1H-pyrazol-5-amine (Compound 18, 47.79 mg, yield 18.94%) was obtained. LCMS: (ESI, m / z): 488.4 [M+H] + .

[0212] 1 H NMR(400MHz,CD3OD)δ9.11-9.10(m,1H),7.88-7.86(m,1H),7.74-7.71(m,1H),7.43(d,J =2.0Hz,1H),6.63(s,1H),6.36(s,1H),6.04(d,J=2.0Hz,1H),4.36(d,J=14.4Hz,1H),3.8 3(d,J=14.4Hz,1H),3.65(s,3H),2.98-2.95(m,3H),2.83-2.68(m,3H),2.63-2.57(m,1H ), 2.41 (d, J = 7.2Hz, 2H), 1.86-1.79 (m, 1H), 1.11 (d, J = 6.0Hz, 3H), 0.89 (d, J = 6.4Hz, 6H).

[0213] Example 19 Preparation of Compound 19

[0214] Compounds Replacement compounds Referring to the synthesis method of compound 13, 5-isobutyl-N-(1-methoxypropan-2-yl)-3-((S)-3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (compound 19, 32.91 mg) was obtained. LCMS: (ESI, m / z): 480.5 [M+H] + .

[0215] 1H NMR (400MHz, CD3OD) δ9.11-9.09(m,1H),7.88-7.86(m,1H),7.74-7.70(m,1H),6.45(d,J=2 0.0Hz,2H),4.34(d,J=14.4Hz,1H),3.80-3.72(m,2H),3.44-3.41(m,1H),3.37-3.33(m,1H ),3.32(s,3H),2.95-2.90(m,3H),2.81-2.70(m,2H),2.67-2.55(m,2H),2.45(d,J=7.2Hz, 2H),1.96-1.85(m,1H),1.20(d,J=6.4Hz,3H),1.09(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H).

[0216] Example 20 Preparation of Compound 20

[0217] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-1-ethyl-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)-1H-pyrazol-4-amine (Compound 20, 42.48 mg, yield 29.86%) was obtained. LCMS: (ESI, m / z): 502.4 [M+H] + .

[0218] 1 H NMR (400MHz, CD3OD) δ9.11-9.09(m,1H),7.89-7.86(m,1H),7.74-7.71(m,1H),7.59(s,1H) ,7.39(s,1H),6.52(d,J=10.8Hz,2H),4.35(d,J=14.4Hz,1H),4.18-4.13(m,2H),3.80(d,J =14.4Hz,1H),2.97-2.95(m,3H),2.82-2.68(m,3H),2.63-2.58(m,1H),2.38(d,J=7.2Hz,2 H),1.87-1.77(m,1H),1.45(t,J=7.2Hz,3H),1.11(d,J=6.0Hz,3H),0.89(d,J=6.4Hz,6H).

[0219] Example 21 Preparation of Compound 21

[0220] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)-1-isopropyl-1H-pyrazol-4-amine (Compound 21, 37.96 mg, yield 29.02%) was obtained. LCMS: (ESI, m / z): 516.5 [M+H] + .

[0221] 1 H NMR(400MHz,CD3OD)δ9.11-9.09(m,1H),7.89-7.86(m,1H),7.74-7.71(m,1H),7.61(s,1H ),7.39(s,1H),6.52(d,J=9.6Hz,2H),4.52-4.45(m,1H),4.35(d,J=14.4Hz,1H),3.81(d,J =14.4Hz,1H),2.97-2.95(m,3H),2.82-2.69(m,3H),2.64-2.59(m,1H),2.38(d,J=7.2Hz,2 H),1.85-1.79(m,1H),1.49(d,J=6.8Hz,6H),1.11(d,J=6.0Hz,3H),0.89(d,J=6.4Hz,6H).

[0222] Example 22 Preparation of Compound 22

[0223] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)-1-isopropyl-1H-pyrazol-4-amine (Compound 22, 78.21 mg, yield 38.99%) was obtained. LCMS: (ESI, m / z): 518.6 [M+H] + .

[0224] 1H NMR (400MHz, CD3OD) δ9.13-9.11(m,1H),7.88-7.86(m,1H),7.76-7.72(m,1H),7.61( s,1H),7.41(s,1H),6.60(s,1H),6.52(s,1H),4.43(d,J=14.4Hz,1H),4.20(t,J=5.6H z,2H),3.92-3.87(m,3H),3.01-2.98(m,3H),2.91-2.88(m,2H),2.78-2.67(m,2H),2 .39(d,J=7.2Hz,2H),1.88-1.80(m,1H),1.15(d,J=6.0Hz,3H),0.90(d,J=6.4Hz,6H).

[0225] Example 23 Preparation of Compound 23

[0226] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(3,3-difluorocyclobutyl)-5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (compound 23, 54.54 mg, yield 50.80%) was obtained. LCMS: (ESI, m / z): 498.4 [M+H] + .

[0227] 1 H NMR(400MHz,CD3OD)δ9.10-9.09(m,1H),7.88-7.86(m,1H),7.74-7.70(m,1H),6.5 1(s,1H),6.28(s,1H),4.33(d,J=14.4Hz,1H),3.94-3.88(m,1H),3.79(d,J=14.4Hz ,1H),3.10-3.00(m,2H),2.95-2.92(m,3H),2.80-2.67(m,3H),2.61-2.55(m,1H), 2.50-2.37(m,4H),1.95-1.85(m,1H),1.10(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H).

[0228] Example 24 Preparation of Compound 24

[0229] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-cyclopentyl-5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (compound 24, 15.80 mg, yield 22.2%) was obtained. LCMS: (ESI, m / z): 476.3 [M+H] + .

[0230] 1 H NMR (400MHz, CD3OD) δ9.13-9.02(m,1H),7.89-7.86(m,1H),7.74-7.70(m,1H),6.44(d,J=8. 4Hz,2H),4.32(d,J=14.4Hz,1H),3.91-3.86(m,1H),3.77(d,J=14.4Hz,1H),2.97-2.94(m,3H ),2.80-2.69(m,3H),2.62-2.56(m,1H),2.46(d,J=7.2Hz,2H),2.04-1.97(m,2H),1.94-1.8 7(m,1H),1.74-1.60(m,4H),1.54-1.47(m,2H),1.11(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H).

[0231] Example 25 Preparation of Compound 25

[0232] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(4,4-difluorocyclohexyl)-5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (Compound 25, 38.70 mg, yield 34.77%) was obtained. LCMS: (ESI, m / z): 526.5 [M+H] + .

[0233] 1H NMR(400MHz,CD3OD)δ9.10-9.09(m,1H),7.89-7.86(m,1H),7.74-7.70(m,1H),6.4 8(d,J=15.6Hz,2H),4.33(d,J=14.4Hz,1H),3.78(d,J=14.4Hz,1H),3.68-3.63(m,1 H),2.98-2.95(m,3H),2.81-2.71(m,3H),2.63-2.57(m,1H),2.46(d,J=7.2Hz,2H), 2.04-1.88(m,7H),1.66-1.59(m,2H),1.11(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H).

[0234] Example 26 Preparation of Compound 26

[0235] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)pyridazin-4-amine (Compound 26, 52.95 mg, 30.16% yield) was obtained. LCMS: (ESI, m / z): 486.1 [M+H] + .

[0236] 1 H NMR(400MHz,CD3OD)δ9.10-9.08(m,1H),8.52(d,J=6.8Hz,1H),8.46(d,J=3.2Hz,1H), 8.19(s,0.26HCOOH),7.83-7.81(m,1H),7.72-7.69(m,1H),7.00-6.97(m,2H),6.85-6. 83(m,1H),4.29(d,J=14.4Hz,1H),3.81(d,J=14.4Hz,1H),2.96-2.85(m,3H),2.75-2. 72(m,1H),2.66-2.56(m,4H),2.46-2.41(m,1H),1.99-1.89(m,1H),1.00-0.95(m,9H).

[0237] Example 27 Preparation of Compound 27

[0238] Compounds Replacement compounds Referring to the synthesis method of compound 3, (S)-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)-1-methyl-1H-pyrazole-4-carboxamide (Compound 27, 33.21 mg, yield 35.81%) was obtained. LCMS: (ESI, m / z): 516.5 [M+H] + .

[0239] 1 H NMR (400MHz, CD3OD) δ9.12 (d, J = 5.2Hz, 1H), 8.10 (s, 1H), 7.92-7.86 (m, 3H), 7. 75-7.72(m,1H),6.95(s,1H),4.40(d,J=14.4Hz,1H),3.94-3.86(m,4H),2.99- 2.97(m,3H),2.88-2.83(m,2H),2.78-2.73(m,1H),2.68-2.63(m,1H),2.56(d, J=7.2Hz,2H),1.99-1.93(m,1H),1.13(d,J=6.0Hz,3H),0.97(d,J=6.4Hz,6H).

[0240] Example 28 Preparation of Compound 28

[0241] Compounds Replacement compounds Referring to the synthesis method of compound 3, a white solid 5-isobutyl-N-(2-methoxycyclopropyl)-3-((S)-3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (compound 28, 1.13 mg, yield 1.58%) was obtained. LCMS: (ESI, m / z): 478.5 [M+H] + .

[0242] 1H NMR (400MHz, CD3OD) δ9.11-9.10(m,1H),7.88-7.86(m,1H),7.74-7.71(m,1H),6.79(d,J=34.4Hz,1H),6. 50(d,J=8.8Hz,1H),4.37(d,J=14.8Hz,1H),3.82(d,J=14.4Hz,1H),3.41(s,3H),3.19-3.17(m,1H),2.95- 2.93(m,3H),2.84-2.79(m,2H),2.74-2.69(m,1H),2.65-2.60(m,1H),2.57-2.54(m,1H),2.50-2.47(m,2 H),1.97-1.90(m,1H),1.11(d,J=6.0Hz,3H),1.08-1.02(m,1H),0.95(d,J=6.4Hz,6H),0.77-0.73(m,1H).

[0243] Example 29 Preparation of Compound 29

[0244] Compounds Replacement compounds Referring to the synthesis method of Compound 5, a white solid (S)-3-((4-(3-((4-fluoro-3-methyl-1H-pyrazol-1-yl)methyl)-5-isobutyl-2-(2H-tetrazol-5-yl)phenyl)-2-methylpiperazin-1-yl)methyl ester)pyridazine (Compound 29, 8.33 mg, yield 11.72%) was obtained. LCMS: (ESI, m / z): 505.8 [M+H] + .

[0245] 1 H NMR(400MHz,CD3OD)δ9.12(d,J=4.8Hz,1H),7.82(d,J=8.4Hz,1H),7.74-7.71(m,1H) ,7.29-7.12(m,1H),7.03-7.01(m,1H),6.82-6.42(m,1H),5.05-5.00(m,2H),4.42-4. 37(m,1H),3.97-3.88(m,1H),2.90-2.81(m,4H),2.71-2.64(m,2H),2.53-2.46(m,3H ),2.09(s,2H),1.97(s,1H),1.91-1.78(m,1H),1.04-1.02(m,3H),0.92-0.87(m,6H).

[0246] Example 30 Preparation of Compound 30

[0247] Compounds Replacement compounds Referring to the synthesis method of Compound 5, a white solid (S)-3-((4-(3-((4-cyclopropyl-1H-pyrazol-1-yl)methyl)-5-isobutyl-2-(2H-tetrazol-5-yl)phenyl)-2-methylpiperazin-1-yl)methyl ester)pyridazine (Compound 30, 17.57 mg, yield 32.07%) was obtained. LCMS: (ESI, m / z): 513.8 [M+H] + .

[0248] 1 H NMR (400MHz, CD3OD) δ9.14 (d, J = 4.8Hz, 1H), 7.82-7.80 (m, 1H), 7.75-7.72 (m, 1H), 7.20 (s, 1H) ),7.03(s,1H),6.90(s,1H),6.79(s,1H),5.10(s,2H),4.43(d,J=15.2Hz,1H),3.98(d,J=11.6 Hz,1H),2.91-2.87(m,4H),2.76-2.67(m,2H),2.57-2.50(m,3H),1.90-1.84(m,1H),1.64-1. 58(m,1H),1.05(d,J=6.0Hz,3H),0.90(d,J=6.8Hz,6H),0.81-0.76(m,2H),0.45-0.41(m,2H).

[0249] Example 31 Preparation of Compound 31

[0250] Compounds Replacement compounds Referring to the synthesis method of Compound 5, a white solid (S)-3-((4-(5-isobutyl-2-(2H-tetrazol-5-yl)-3-(4-(trifluoromethyl)-1H-pyrazol-1-yl)methyl)phenyl)-2-methylpiperazin-1-yl)methyl ester)pyridazine (Compound 31, 33.97 mg, yield 51.99%) was obtained. LCMS: (ESI, m / z): 541.5 [M+H] + .

[0251] 1H NMR(400MHz,CD3OD)δ9.10(d,J=3.2Hz,1H),7.83–7.80(m,1H),7.73–7.70(m,1H), 7.65(d,J=3.6Hz,2H),7.01(s,1H),6.85(s,1H),5.19(s,2H),4.32(d,J=14.0Hz,1H ),3.85(d,J=16.4Hz,1H),2.90-2.86(m,3H),2.78–2.74(m,1H),2.66–2.62(m,2H), 2.52–2.41(m,3H),1.90–1.83(m,1H),1.00(d,J=5.6Hz,3H),0.90(d,J=6.8Hz,6H).

[0252] Example 32 Preparation of Compound 32

[0253] Compounds Replacement compounds Referring to the synthesis method of compound 3, a white solid (S)-N-cyclobutyl-5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)aniline (compound 32, 28.66 mg, yield 12.10%) was obtained. LCMS: (ESI, m / z): 462.7 [M+H] + .

[0254] 1 H NMR(400MHz,CD3OD)δ9.11(d,J=4.8Hz,1H),7.89-7.86(m,1H),7.74-7.71(m,1H) ,6.44(s,1H),6.31(s,1H),4.40-4.35(m,1H),4.01-3.94(m,1H),3.87-3.80(m,1 H),2.97-2.95(m,3H),2.86-2.81(m,2H),2.76-2.71(m,1H),2.67-2.62(m,1H),2 .47-2.41(m,4H),1.93-1.82(m,5H),1.13(d,J=6.0Hz,3H),0.93(d,J=6.4Hz,6H).

[0255] Example 33 Preparation of Compound 33

[0256] Compounds Replacement compounds Referring to the synthesis method of compound 4, a white solid (S)-N-(5-isobutyl-3-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)-2-(2H-tetrazol-5-yl)phenyl)bicyclo[1.1.1]pentan-1-amine (compound 33, 7.60 mg, yield 6.95%) was obtained. LCMS: (ESI, m / z): 474.5 [M+H] + .

[0257] 1 H NMR (400MHz, CD3OD) δ9.27 (s, 1H), 7.83 (d, J = 3.2Hz, 2H), 6.80 (s, 1H), 6.4 9(s,1H),5.00-4.91(m,1H),4.62-4.53(m,1H),3.60-3.53(m,1H),3.50-3. 45(m,1H),3.15-3.06(m,3H),2.98-2.93(m,1H),2.50(s,1H),2.48(s,2H) ,2.10(s,6H),1.97-1.88(m,1H),1.39-1.29(m,4H),0.96(d,J=6.8Hz,6H).

[0258] Example 34 Preparation of Compound 34

[0259] The preparation method of reference compound 3 was used replace After two steps of reaction, column chromatography was performed to obtain compound 34, 26.47 mg, in a yield of 21.92%.

[0260] LCMS: (ESI, m / z): 484.4 [M+H] + .

[0261] 1 H NMR (400MHz, DMSO-d6) δ9.11–9.10 (m, 1H), 7.64 (d, J = 3.2Hz, 2H), 6.53 (s, 1H), 6.24(s,1H),4.01(d,J=14.0Hz,1H),3.71–3.65(m,4H),3.56(d,J=14.0Hz,1H) ,2.78–2.71(m,2H),2.66–2.62(m,1H),2.43(d,J=6.8Hz,4H),2.24(s,1H),2.0 1–1.96(m,1H),1.88–1.85(m,1H),0.90(d,J=6.8Hz,6H),0.83(d,J=6.4Hz,3H).

[0262] Example 35 Preparation of Compound 35

[0263] The preparation method of reference compound 3 was used replace After two steps of reaction, column chromatography was performed to obtain compound 35, 48.68 mg, in a yield of 37.10%.

[0264] LCMS: (ESI, m / z): 498.3 [M+H] + .

[0265] 1 H NMR (400MHz, DMSO-d6) δ16.16 (s, 1H), 9.10 (s, 1H), 7.65 (d, J = 2.8Hz, 2H), 6.62 (s,1H),6.55(s,1H),4.02(d,J=14.4Hz,1H),3.56(d,J=14.0Hz,1H),3.10–2.9 8(m,4H),2.80(d,J=10.0Hz,1H),2.72–2.67(m,2H),2.45(d,J=6.8Hz,4H),2.3 3–2.21(m,3H),1.95–1.85(m,2H),0.90(d,J=6.4Hz,6H),0.85(d,J=5.6Hz,3H).

[0266] Example 36 Preparation of Compound 36

[0267] The preparation method of reference compound 2 was used replace After three steps of reaction, column chromatography gave compound 36, 50.30 mg, with a yield of 32.68%. LCMS: (ESI, m / z): 479.4 [M+H] + .

[0268] 1 H NMR(400MHz,CD3OD)δ9.08–9.07(m,1H),7.82(d,J=8.4Hz,1H),7.71–7.68(m,1H),6 .65(d,J=8.4Hz,2H),5.01(s,1H),4.23(d,J=14.0Hz,1H),3.90–3.87(m,1H),3.78–3 .70(m,3H),3.68–3.62(m,1H),2.88–2.80(m,3H),2.69–2.61(m,2H),2.53(d,J=7.2 Hz,3H),2.34–2.30(m,1H),2.17–2.08(m,1H),1.94–1.89(m,2H),0.99–0.93(m,9H).

[0269] Example 37 Preparation of Compound 37

[0270] The preparation method of reference compound 2 was used replace After three steps of reaction, column chromatography gave compound 37, 22.74 mg, 56% yield. LCMS: (ESI, m / z): 499.4 [M+H] + .

[0271] 1 H NMR(400MHz,MeOD)δ9.08(dd,J=4.9,1.6Hz,1H),7.82(dd,J=8.5,1.6Hz,1H),7.70(dd,J=8.5,4 .9Hz,1H),6.66(s,1H),6.51(s,1H),4.69(s,1H),4.21(d,J=14.4Hz,1H),3.73(d,J=14.3Hz,1H) ,3.02(td,J=13.0,6.8Hz,2H),2.83(dd,J=27.0,9.8Hz,3H),2.65(d,J=11.7Hz,1H),2.61–2.43 (m,6H),2.31(s,1H),1.91(dt,J=13.6,6.9Hz,1H),0.97(d,J=6.2Hz,3H),0.93(d,J=6.6Hz,6H).

[0272] Example 38 Preparation of Compound 104

[0273] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 104 (142.88 mg, yield 31.79%) was obtained by column chromatography.

[0274] LCMS: (ESI, m / z): 488.4 [M+H] + .

[0275] 1H NMR (400MHz, CD3OD) δ9.10(d,J=4.4Hz,1H),7.82(d,J=8.4Hz,1H),7.73–7.70(m,1H),6.49(d,J= 10.0Hz,2H),4.27(d,J=14.4Hz,1H),3.78(d,J=14.4Hz,1H),3.03–2.99(m,2H),2.96–2.93(m,1H ),2.86–2.84(m,2H),2.70–2.65(m,3H),2.62–2.57(m,1H),2.47–2.45(m,3H),2.27(m,1H),1.93 –1.86(m,1H),1.76–1.64(m,2H),1.01(d,J=6.0Hz,3H),0.93(d,J=8.4Hz,6H),0.51–0.43(m,4H).

[0276] Example 39 Preparation of Compound 106

[0277] Step 1: Synthesis of compound 106-2

[0278] To a solution of 3-oxocyclobutane-1-carboxylic acid (2 g, 17.53 mmol, 1.0 equiv) in dichloromethane (30 mL) was added 2-hydroxyisoindole-1,3-dione (2.86 g, 17.53 mmol, 1.0 equiv), 4-dimethylaminopyridine (2.14 g, 17.53 mmol, 1.0 equiv), and N,N'-dicyclohexylcarbodiimide (5.43 g, 26.29 mmol, 1.5 equiv) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-50%) to obtain yellow solid compound 106-2 (3.6 g, yield 79.23%).

[0279] 1 H NMR (400MHz, CDCl3) δ7.92-7.90(m,2H),7.83-7.81(m,2H),3.69–3.50(m,5H).

[0280] Step 2: Synthesis of compound 106-3

[0281] To a solution of compound 106-2 (150 mg, 0.578 mmol, 1.0 equiv) in N,N-dimethylacetamide (2 mL) in a microwave tube was added (S)-2-bromo-4-isobutyl-6-(3-methyl-4-(pyridazin-3-ylmethyl)piperazin-1-yl)benzonitrile (250 mg, 0.578 mmol, 1.0 equiv), 6,6'-di-tert-butyl-2,2'-bipyridine (31 mg, 0.115 mmol, 0.2 equiv), and nickel chloride (15 mg, 0.115 mmol, 0.2 equiv). The reaction mixture was stirred at room temperature for 2 days. After completion of the reaction, the reaction mixture was quenched by addition of ice water (10 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to give yellow solid compound 106-3 (19 mg, yield 7.86%).

[0282] LCMS: (ESI, m / z): 418.3 [M+H] + .

[0283] Step 3: Synthesis of Compound 106-4

[0284] To a solution of compound 106-3 (9.0 mg, 0.045 mmol, 1.0 equiv) in dichloromethane (2 mL) was added diethylaminosulfur trifluoride (3.4 mg, 0.091 mmol, 2.0 equiv) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (5 mL) and extracted with ethyl acetate (5 mL x 3). The combined organic layers were washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford compound 106-4 (9 mg, 45.0% yield) as a yellow solid.

[0285] LCMS: (ESI, m / z): 440.5 [M+H] + .

[0286] Step 4: Synthesis of Compound 106

[0287] Referring to the preparation method of tetrazole in compound 2, compound 106 (1.54 mg, yield 15.59%) was obtained.

[0288] LCMS: (ESI, m / z): 483.3 [M+H] + .

[0289] 1H NMR (400MHz, CD3OD) δ9.09 (s, 1H), 8.36 (s, 0.65HCOOH), 7.82 (d, J = 7.6Hz, 1H), 7.72-7.69 (m ,1H),6.94(d,J=16.8Hz,2H),4.26(d,J=14.4Hz,1H),3.80(d,J=15.2Hz,1H),3.20-3.13(m, 1H),2.92-2.81(m,3H),2.73-2.69(m,1H),2.65-2.62(m,1H),2.56-2.52(m,3H),2.50-2.41 (m,4H),2.38-2.32(m,1H),1.94-1.88(m,1H),0.97(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H).

[0290] Example 40 Preparation of Compound 121

[0291] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 121 (18.33 mg, yield 21%) was obtained by column chromatography.

[0292] LCMS: (ESI, m / z): 524.4 [M+H] + .

[0293] 1 H NMR(400MHz,CD3OD)δ9.10(d,J=4.8Hz,1H),7.81(d,J=8.8Hz,1H),7.73–7.69(m,1H ),6.48(s,1H),6.17(s,1H),4.28(d,J=14.4Hz,1H),3.81(d,J=14.0Hz,1H),3.42(d, J=4.0Hz,4H),2.91–2.85(m,3H),2.72–2.69(m,1H),2.65–2.59(m,5H),2.52–2.44( m,3H),2.33(s,1H),1.90–1.85(m,1H),0.99(d,J=6.4Hz,3H),0.92(d,J=6.8Hz,6H).

[0294] Example 41 Preparation of Compound 129

[0295] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 129 (45.14 mg, 48.4% yield) was obtained by column chromatography.

[0296] LCMS: (ESI, m / z): 502.3 [M+H] + .

[0297] 1 H NMR (400MHz, CD3OD) δ9.07(d,J=4.8Hz,1H),7.83(d,J=8.4Hz,1H),7.72–7.68(m,1H),6.77(d, J=17.6Hz,2H),4.20(d,J=14.0Hz,1H),3.70(d,J=14.4Hz,1H),2.97–2.93(m,1H),2.91–2.88(m ,1H),2.86–2.83(m,5H),2.65–2.59(m,2H),2.51(d,J=7.2Hz,2H),2.44(s,1H),2.25–2.22(m, 1H),1.93–1.86(m,1H),1.17(s,4H),0.98(d,J=6.4Hz,3H),0.93(d,J=6.4Hz,6H),0.24(s,4H).

[0298] Example 42 Preparation of Compound 131

[0299] Step 1: Synthesis of compound 131-1

[0300] To a solution of (S)-tert-butyl 4-(3-bromo-2-cyano-5-isobutylphenyl)-2-methylpiperazine-1-carboxylate (150.0 mg, 0.343 mmol, 1.0 equiv) in 1,4-dioxane (5 mL) and water (1 mL) in a microwave tube was added 2-(4,4-difluorocyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (101.0 mg, 0.412 mmol, 1.2 equiv), 1,1-bis(diphenylphosphino)diphenylferric palladium chloride (25 mg, 0.0346 mmol, 0.1 equiv), and potassium carbonate (143 mg, 1.0 mmol, 3.0 equiv). The reaction mixture was heated to 90° C. under nitrogen and stirred for 16 hours. After the reaction was completed, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-50%) to obtain compound 131-1 (123.0 mg, yield 75.56%) as a yellow solid. LCMS: (ESI, m / z): 474.2 [M+H] + .

[0301] Step 2: Synthesis of compound 131-2

[0302] To a solution of compound 131-1 (123.0 mg, 0.26 mmol, 1.0 equiv) in methanol (3 mL) was added palladium on carbon (20 mg) at room temperature to displace the hydrogen atmosphere. The reaction mixture was heated to 70°C and stirred for 3 hours. After the reaction, the reaction mixture was filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-50%) to afford compound 131-2 (100.0 mg, 80.96% yield) as a yellow solid.

[0303] LCMS: (ESI, m / z): 476.1 [M+H] + .

[0304] Step 3: Synthesis of compound 131-3

[0305] To a solution of compound 131-2 (100.0 mg, 0.21 mmol, 1.0 equiv) in dichloromethane (5 mL) was added a solution of hydrochloric acid in 1,4-dioxane (2 mg) at room temperature. The reaction mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was concentrated to afford the hydrochloride salt of compound 131-3 (110.0 mg, crude product) as a yellow solid.

[0306] LCMS: (ESI, m / z): 376.4 [M+H] + .

[0307] Step 4: Synthesis of compound 131-4

[0308] To a solution of compound 131-3 hydrochloride (110.0 mg, 0.26 mmol, 1.0 equiv) in N,N-dimethylformamide (3 mL) at room temperature were added N,N-diisopropylethylamine (104.0 mg, 0.8 mmol, 3.0 equiv) and 3-(chloromethyl)pyridazine (41 mg, 0.32 mmol, 1.2 equiv). The reaction mixture was heated to 50°C and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford compound 131-4 (76.0 mg, 60.87% yield) as a yellow solid.

[0309] LCMS: (ESI, m / z): 468.6 [M+H] + .

[0310] Step 5: Synthesis of Compound 131

[0311] Referring to the preparation method of tetrazole in compound 2, compound 131 (34.88 mg, yield 42.03%) was obtained.

[0312] LCMS: (ESI, m / z): 511.3 [M+H] + .

[0313] 1 H NMR(400MHz,CD3OD)δ9.11(d,J=3.6Hz,1H),8.18(s,0.13HCOOH),7.81(d,J=7.2Hz,1H),7.74-7.70(m,1 H),6.94(d,J=17.6Hz,2H),4.32(d,J=14.4Hz,1H),3.86(d,J=14.4Hz,1H),2.93-2.84(m,3H),2.78-2.7 4(m,1H),2.68-2.63(m,1H),2.60-2.52(m,3H),2.41-2.36(m,1H),2.28-2.21(m,1H),2.06-1.99(m,2H) ,1.92-1.85(m,1H),1.80-1.71(m,4H),1.68-1.52(m,2H),1.00(d,J=6.0Hz,3H),0.93(d,J=6.4Hz,6H).

[0314] Example 43 Preparation of Compound 132

[0315] Step 1: Synthesis of compound 132-1

[0316] Under nitrogen protection, a solution of (S)-tert-butyl 4-(3-bromo-2-cyano-5-isobutylphenyl)-2-methylpiperazine-1-carboxylate (200.0 mg, 0.460 mmol, 1.0 equiv) in 1,4-dioxane / water (5.0 mL / 1.0 mL) was added to a microwave tube. Then, 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (145.0 mg, 0.690 mmol, 1.5 equiv), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (38 mg, 0.046 mmol, 0.1 equiv), and cesium carbonate (450 mg, 1.379 mmol, 3.0 equiv) were added. The reaction mixture was heated to 90°C and reacted for 16 hours. The desired product was found in the liquid phase. The reaction mixture was cooled to room temperature, filtered, and the filtrate was extracted with ethyl acetate (100 mL). The organic layer was washed with water (2 × 50 mL) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (100%) to obtain compound 132-1 (190 mg, 94.09%) as a yellow oil. LCMS: (ESI, m / z): 440.1 [M+H] + .

[0317] Step 2: Synthesis of compound 132-2

[0318] Compound 132-1 (100.0 mg, 0.228 mmol, 1.0 equiv) was dissolved in methanol (10 mL), and palladium on carbon (20.0 mg, 10%) was added. The air was replaced, hydrogen was introduced, and the reaction was carried out at 70°C for 3 hours. Celite was then added for filtration, and the filtrate was concentrated under reduced pressure to obtain compound 132-2 (85 mg, 84.92%) as a colorless oil.

[0319] LCMS: (ESI, m / z): 442.4 [M+H] + .

[0320] Step 3: Synthesis of compound 132-3

[0321] Under nitrogen, hydrochloric acid / dioxane (4 mL, 4 mol / L) was added to a solution of compound 132-2 (85.0 mg, 0.193 mmol, 1.0 equiv) in dichloromethane (4 mL) at room temperature. The mixture was allowed to react for 3 hours. The desired product was observed in the liquid phase. The solution was concentrated under reduced pressure to afford the hydrochloride salt of compound 132-3 (92 mg, crude) as a white solid.

[0322] LCMS: (ESI, m / z): 342.1 [M+H] + .

[0323] Step 4: Synthesis of compound 132-4

[0324] Under nitrogen, to a solution of compound 132-3 hydrochloride (92.0 mg, 0.244 mmol, 1.0 equiv) in N,N-dimethylformamide (5 mL) were added 3-(chloromethyl)pyridazine hydrochloride (80.0 mg, 0.488 mmol, 2.0 equiv) and N,N-diisopropylethylamine (157.0 mg, 1.220 mmol, 5.0 equiv), respectively. The reaction was allowed to react at room temperature for 16 hours, and the desired product was observed in the liquid. The reaction solution was poured into water (20 mL). The system was extracted with ethyl acetate (3 x 20 mL). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to dryness. The resulting residue was directly purified by column chromatography with methanol / dichloromethane (10%) to afford compound 132-4 (45 mg, 40.59%) as a yellow solid.

[0325] LCMS: (ESI, m / z): 434.4 [M+H] + .

[0326] Step 5: Synthesis of compound 132

[0327] Referring to the preparation method of tetrazole in compound 2, compound 132 (11.83 mg, 23.90%) was obtained.

[0328] LCMS: (ESI, m / z): 477.3 [M+H] + .

[0329] 1 H NMR (400MHz, CD3OD) δ9.10(d,J=3.6Hz,1H),7.82(d,J=7.2Hz,1H),7.73–7.70(m,1H),7.00(s ,1H),6.92(s,1H),4.30(d,J=14.4Hz,1H),3.93–3.82(m,3H),3.31–3.22(m,2H),2.93–2.84(m ,3H),2.76–2.71(m,1H),2.67–2.62(m,1H),2.54(d,J=7.2Hz,3H),2.42–2.36(m,2H),1.93–1. 86(m,1H),1.81–1.71(m,2H),1.62–1.56(m,2H),0.99(d,J=6.0Hz,3H),0.93(d,J=6.4Hz,6H).

[0330] Example 44 Preparation of Compound 133

[0331] Step 1: Synthesis of compound 133-1

[0332] To a solution of compound 3-1 (200.0 mg, 0.466 mmol, 1.0 equiv) in 1,4-dioxane (5 mL) and water (1 mL) in a microwave tube was added 2-(4,4-difluorocyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (114.0 mg, 0.466 mmol, 1.0 equiv), 1,1-bis(diphenylphosphino)diborane iron palladium dichloride (35 mg, 0.046 mmol, 0.1 equiv), and potassium carbonate (194 mg, 1.4 mmol, 3.0 equiv). The reaction mixture was heated to 90°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to give yellow solid compound 133-1 (220.0 mg, crude product).

[0333] LCMS: (ESI, m / z): 466.3 [M+H] + .

[0334] Step 2: Synthesis of compound 133

[0335] Referring to the preparation method of tetrazole in compound 2, compound 133 (51.63 mg, yield 21.48%) was obtained.

[0336] LCMS: (ESI, m / z): 509.2 [M+H] + .

[0337] 1H NMR(400MHz,CD3OD)δ9.10(d,J=5.2Hz,1H),8.14(s,0.16HCOOH),7.82(d,J=8.4Hz,1H),7.73 -7.70(m,1H),6.99(s,1H),6.87(s,1H),5.25(s,1H),4.29(d,J=14.4Hz,1H),3.82(d,J=14.4 Hz,1H),2.93-2.87(m,3H),2.75-2.71(m,1H),2.68-2.61(m,1H),2.59-2.50(m,3H),2.46-2. 31(m,3H),2.26-2.22(m,2H),1.94-1.86(m,3H),1.00(d,J=6.0Hz,3H),0.93(d,J=6.4Hz,6H).

[0338] Example 45 Preparation of Compound 135

[0339] Step 1: Synthesis of compound 135-1

[0340] To a solution of (S)-tert-butyl 4-(3-bromo-2-cyano-5-isobutylphenyl)-2-methylpiperazine-1-carboxylate (150.0 mg, 0.343 mmol, 1.0 equiv) in 1,4-dioxane (5 mL) and water (1 mL) in a microwave tube was added 2-(2,5-dihydrofuran-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (81.0 mg, 0.412 mmol, 1.2 equiv), 1,1-bis(diphenylphosphino)diphenylferric palladium chloride (25 mg, 0.0346 mmol, 0.1 equiv), and potassium carbonate (143 mg, 1.0 mmol, 3.0 equiv). The reaction mixture was heated to 90° C. under nitrogen and stirred for 16 hours. After the reaction was completed, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-50%) to obtain compound 135-1 (105.0 mg, yield 71.78%).

[0341] LCMS: (ESI, m / z): 426.2 [M+H] + .

[0342] Step 2: Synthesis of compound 135-2

[0343] To a solution of compound 135-1 (105.0 mg, 0.246 mmol, 1.0 equiv) in methanol (3 mL) was added palladium on carbon (20 mg) at room temperature to displace the hydrogen atmosphere. The reaction mixture was heated to 70°C and stirred for 3 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-50%) to afford compound 135-2 (92.0 mg, 87.21% yield).

[0344] LCMS: (ESI, m / z): 428.3 [M+H] + .

[0345] Step 3: Synthesis of compound 135-3

[0346] To a solution of compound 135-2 (92.0 mg, 0.21 mmol, 1.0 equiv) in dichloromethane (5 mL) was added a solution of hydrochloric acid in 1,4-dioxane (2 mg) at room temperature. The reaction mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was concentrated to afford compound 135-3 (105.0 mg, crude product).

[0347] LCMS: (ESI, m / z): 328.3 [M+H] + .

[0348] Step 4: Synthesis of compound 135-4

[0349] To a solution of compound 135-3 (105.0 mg, 0.288 mmol, 1.0 equiv) in N,N-dimethylformamide (3 mL) were added N,N-diisopropylethylamine (112.0 mg, 0.86 mmol, 3.0 equiv) and 3-(chloromethyl)pyridazine (37 mg, 0.288 mmol, 1.0 equiv) at room temperature. The reaction mixture was heated to 50°C and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford compound 135-4 (62.0 mg, 51.22% yield).

[0350] LCMS: (ESI, m / z): 420.3 [M+H] + .

[0351] Step 5: Synthesis of Compound 135

[0352] Referring to the preparation method of tetrazole in compound 2, compound 135 (20.02 mg, yield 29.29%) was obtained.

[0353] LCMS: (ESI, m / z): 463.3 [M+H] + .

[0354] 1 H NMR(400MHz,CD3OD)δ9.11(d,J=3.6Hz,1H),8.13(s,0.29HCOOH),7.81(d,J=8.4Hz,1H),7.74-7.71(m,1H),7 .06(s,1H),6.94(s,1H),4.34(d,J=14.4Hz,1H),4.00-3.96(m,1H),3.91-3.81(m,2H),3.76-3.70(m,1H),3. 68-3.65(m,1H),3.04-2.97(m,1H),2.94-2.85(m,3H),2.80-2.76(m,1H),2.69-2.59(m,2H),2.54(d,J=7.2H z,2H),2.45-2.40(m,1H),2.21-2.11(m,1H),2.00-1.87(m,2H),1.01(d,J=6.0Hz,3H),0.93(d,J=6.4Hz,6H).

[0355] Example 46 Preparation of Compound 136

[0356] The preparation method of reference compound 137 was used replace After two steps of reaction, compound 136 (9.15 mg, yield 7.93%) was obtained by column chromatography.

[0357] LCMS: (ESI, m / z): 476.1 [M+H] + .

[0358] 1H NMR(400MHz,CD3OD)δ9.11–9.09(m,1H),7.86–7.83(m,1H),7.73–7.70(m,1H),7.58 (s,1H),6.88(s,1H),4.33(d,J=14.4Hz,1H),3.81(d,J=14.8Hz,1H),2.91–2.86(m,3 H),2.77(d,J=11.6Hz,1H),2.66–2.61(m,2H),2.53–2.46(m,3H),1.95–1.88(m,1H) ,1.62–1.56(m,1H),1.05(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H),0.84–0.78(m,4H).

[0359] Example 47 Preparation of Compound 137

[0360] Step 1: Synthesis of compound 137-1

[0361] To a solution of compound 3-1 (100.0 mg, 0.234 mmol, 1.0 equiv) in 1,4-dioxane (5 mL) in a microwave tube was added 3,3-difluorocyclobutane-1-carboxamide (47.0 mg, 0.351 mmol, 1.5 equiv), (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (20 mg, 0.023 mmol, 0.1 equiv), and cesium carbonate (229 mg, 0.703 mmol, 3.0 equiv). The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to give compound 137-1 (102.0 mg, 90.44%).

[0362] LCMS: (ESI, m / z): 483.3 [M+H] + .

[0363] Step 2: Synthesis of Compound 137

[0364] Referring to the preparation method of tetrazole in compound 2, compound 137 (6.08 mg, yield 5.46%) was obtained.

[0365] LCMS: (ESI, m / z): 526.2 [M+H] + .

[0366] 1 H NMR(400MHz,CD3OD)δ9.11–9.09(m,1H),7.85–7.83(m,1H),7.74–7.70(m,1H) ,7.47(s,1H),6.92(s,1H),4.34(d,J=14.4Hz,1H),3.82(d,J=14.8Hz,1H),2.9 7–2.93(m,1H),2.89–2.85(m,3H),2.80–2.75(m,2H),2.71-2.63(m,5H),2.55 –2.53(m,3H),1.96–1.89(m,1H),1.05(d,J=5.6Hz,3H),0.94(d,J=6.8Hz,6H).

[0367] Example 48 Preparation of Compound 165

[0368] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 165 (23.05 mg, yield 27.78%) was obtained by column chromatography.

[0369] LCMS: (ESI, m / z): 510.3 [M+H] + .

[0370] 1 H NMR(400MHz,CD3OD)δ9.09(d,J=4.8Hz,1H),7.81(d,J=8.4Hz,1H),7.72–7.69(m,1H),6.5 5(d,J=19.6Hz,2H),4.25(d,J=14.4Hz,1H),3.77(d,J=15.6Hz,1H),3.21(d,J=9.2Hz,2H) ,3.13–3.06(m,2H),2.99–2.93(m,1H),2.86–2.84(m,2H),2.68–2.58(m,2H),2.48–2.41( m,3H),2.28–2.22(m,3H),1.92–1.86(m,1H),1.00(d,J=6.4Hz,3H),0.93(d,J=6.4Hz,6H).

[0371] Example 49 Preparation of Compound 173

[0372] Reference is made to the preparation method of compound 3, using 179-5 instead of 3-1, and using replace After two steps of reaction, compound 173 (4.62 mg, yield 4.25%) was obtained by column chromatography.

[0373] LCMS: (ESI, m / z): 532.2 [M+H] + .

[0374] 1 H NMR(400MHz,CD3OD)δ9.09(d,J=3.2Hz,1H),7.83-7.80(m,1H),7.72-7.69(m,1H),6.68(d,J =33.6Hz,2H),4.26(d,J=13.6Hz,1H),3.79(d,J=13.6Hz,1H),3.16-3.12(m,2H),2.98(d,J=3 .2Hz,1H),2.96-2.87(m,4H),2.81-2.77(m,2H),2.70-2.59(m,2H),2.51-2.46(m,1H),2.41- 2.15(m,3H),1.92-1.85(m,1H),1.56-1.48(m,1H),1.20-1.13(m,1H),1.00(d,J=6.4Hz,3H).

[0375] Example 50 Preparation of Compound 179

[0376] Step 1: Synthesis of compound 179-2

[0377] To a solution of 2-amino-4-bromo-6-fluorobenzonitrile (5 g, 23.25 mmol, 1.0 equiv) in 1,4-dioxane (30 mL) and water (6 mL) was added 2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (4.69 g, 27.90 mmol, 1.2 equiv), 1,1-bis(diphenylphosphino)diborane iron palladium chloride (1.7 g, 2.33 mmol, 0.1 equiv), and potassium carbonate (9.64 g, 69.76 mmol, 3.0 equiv) at room temperature. The reaction mixture was heated to 90°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-50%) to give compound 179-2 (6 g, crude) as a yellow oil.

[0378] LCMS: (ESI, m / z): 177.1 [M+H] + .

[0379] Step 2: Synthesis of compound 179-3

[0380] To a solution of 4-allyl-2-amino-6-fluorobenzonitrile (6 g, 34.05 mmol, 1.0 equiv) in acetonitrile (50 mL) was added cuprous bromide (9.77 g, 68.11 mmol, 2.0 equiv) and tert-butyl nitrite (10.53 g, 102.16 mmol, 3.0 equiv) at room temperature. The reaction mixture was heated to 40°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of ice water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-10%) to afford compound 179-3 (1.99 g, yield: 24.34%) as a yellow oil.

[0381] 1 H NMR (400MHz, CDCl3) δ7.34 (s, 1H), 7.02 (d, J = 9.2Hz, 1H), 5.91-5.84 (mz, 1H), 5.23-5.14 (m, 2H), 3.42 (d, J = 6.8Hz, 2H).

[0382] Step 3: Synthesis of Compound 179-4

[0383] To a solution of compound 179-3 (1.99 g, 8.29 mmol, 1.0 equiv) in toluene (10 mL) at room temperature was added tetrabutylammonium bromide (267.2 mg, 0.829 mmol, 0.1 equiv) and (bromodifluoromethyl)trimethylsilane (8.42 g, 41.45 mmol, 5.0 equiv) in a microwave oven. The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was concentrated, and the resulting residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-10%) to afford compound 179-4 (1.2 g, yield: 49.91%) as a yellow oil.

[0384] 1 H NMR (400MHz, CDCl3) δ7.38 (s, 1H), 7.08 (d, J = 9.2Hz, 1H), 2.90-2.79 (m, 2H), 1.82-1.71 (m, 1H), 1.64-1.57 (m, 1H), 1.17-1.09 (m, 1H).

[0385] Step 4: Synthesis of Compound 179-5

[0386] To a solution of compound 179-4 (350 mg, 1.21 mmol, 1.0 equiv) in dimethyl sulfoxide (5 mL) at room temperature were added N,N-diisopropylethylamine (468 mg, 3.63 mmol, 3.0 equiv) and (S)-3-((2-methylpiperazin-1-yl)methyl)pyridazine hydrochloride (331 mg, 1.45 mmol, 1.2 equiv). The reaction mixture was heated to 120°C under nitrogen and stirred for 3 hours. After completion of the reaction, the reaction mixture was quenched by addition of ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford compound 179-5 (280 mg, crude) as a yellow solid.

[0387] LCMS: (ESI, m / z): 461.9 [M+H] + .

[0388] Step 5: Synthesis of Compound 179-6

[0389] To a solution of compound 179-5 (240 mg, 0.519 mmol, 1.0 equiv) in 1,4-dioxane (5 mL) was added 3,3-difluorocyclobutane-1-amine (67 mg, 0.62 mmol, 1.2 equiv), methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) (44 mg, 0.0519 mmol, 0.1 equiv), and cesium carbonate (508 mg, 1.56 mmol, 3.0 equiv) at room temperature. The reaction mixture was heated to 80°C under nitrogen and stirred for 2 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to give yellow solid compound 179-6 (70 mg, yield: 27.60%).

[0390] LCMS: (ESI, m / z): 489.5 [M+H] + .

[0391] Step 6: Synthesis of Compound 179

[0392] Referring to the preparation method of tetrazole in compound 2, compound 179 (28.03 mg, yield 36.80%) was obtained.

[0393] LCMS: (ESI, m / z): 532.3 [M+H] + .

[0394] 1 H NMR (400MHz, CD3OD) δ9.10-9.09(m,1H),7.87-7.85(m,1H),7.74-7.70(m,1H),6.58(s ,1H),6.38(s,1H),4.33(d,J=14.4Hz,1H),3.92-3.87(m,1H),3.78(d,J=14.4Hz,1H),3 .09-3.02(m,2H),2.92-2.90(m,3H),2.79-2.64(m,5H),2.58-2.55(m,1H),2.49-2.39( m,2H),1.93-1.80(m,1H),1.55-1.52(m,1H),1.20-1.16(m,1H),1.09(d,J=6.0Hz,3H).

[0395] Example 51 Preparation of Compound 184-A

[0396] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 184-A (35.47 mg, yield 36.10%) was obtained by column chromatography.

[0397] LCMS: (ESI, m / z): 512.4 [M+H] + .

[0398] 1H NMR(400MHz,CD3OD)δ9.09(d,J=4.0Hz,1H),8.35(s,0.24HCOOH),7.87(d,J=7.6Hz,1H),7.73- 7.70(m,1H),6.44(d,J=24.0Hz,2H),4.32(d,J=14.4Hz,1H),4.11-4.06(m,1H),3.77(d,J=14.4 Hz,1H),2.95-2.93(m,3H),2.79-2.67(m,3H),2.61-2.53(m,2H),2.47(d,J=7.2Hz,2H),2.31-2 .07(m,3H),1.98-1.88(m,2H),1.78-1.72(m,1H),1.09(d,J=5.6Hz,3H),0.94(d,J=6.4Hz,6H).

[0399] Example 52 Preparation of Compound 184-B

[0400] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 184-B (43.25 mg, yield 44.01%) was obtained by column chromatography.

[0401] LCMS: (ESI, m / z): 512.3 [M+H] + .

[0402] 1 H NMR(400MHz,CD3OD)δ9.09(d,J=4.8Hz,1H),7.87(d,J=8.2Hz,1H),7.73-7.70(m,1H),6.4 4(d,J=23.6Hz,2H),4.32(d,J=14.4Hz,1H),4.11-4.08(m,1H),3.77(d,J=14.4Hz,1H),2. 95-2.92(m,3H),2.79-2.66(m,3H),2.62-2.52(m,2H),2.47(d,J=7.2Hz,2H),2.31-2.06( m,3H),1.96-1.88(m,2H),1.79-1.72(m,1H),1.09(d,J=6.0Hz,3H),0.94(d,J=6.4Hz,6H).

[0403] Example 53 Preparation of Compound 185

[0404] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 185 (32.09 mg, yield 30.72%) was obtained by column chromatography.

[0405] LCMS: (ESI, m / z): 476.3 [M+H] + .

[0406] 1 H NMR (400MHz, CD3OD) δ9.09 (s, 1H), 7.87 (d, J = 7.6Hz, 1H), 7.73-7.70 (m, 1H), 6.32 (d, J=66.0Hz,2H),4.33(d,J=14.4Hz,1H),3.77(d,J=14.4Hz,1H),2.98-2.91(m,3H),2. 80-2.68(m,3H),2.61-2.56(m,1H),2.43-2.40(m,2H),2.23-2.19(m,2H),2.15-2.09 (m,2H),1.97-1.84(m,3H),1.48(s,3H),1.10(d,J=4.8Hz,3H),0.93(d,J=5.6Hz,6H).

[0407] Example 54 Preparation of Compound 196

[0408] Step 1: Synthesis of Compound 196-1

[0409] To a solution of compound 3-1 (500.0 mg, 1.17 mmol, 1.0 equiv) in 1,4-dioxane (10 mL) in a microwave tube was added benzophenone imine (212 mg, 1.17 mmol, 1.0 equiv), (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (98 mg, 0.117 mmol, 0.1 equiv), and cesium carbonate (1.14 g, 3.5 mmol, 3.0 equiv). The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to give compound 196-1 (600.0 mg, 97.23%).

[0410] LCMS: (ESI, m / z): 529.4 [M+H] + .

[0411] Step 2: Synthesis of compound 196-2

[0412] Under nitrogen, sodium azide (369.0 mg, 5.67 mmol, 5 equiv) and tri-n-butyltin azide (1.85 g, 5.67 mmol, 5 equiv) were added to a toluene (6 mL) solution of compound 196-1 (600.0 mg, 1.13 mmol, 1.0 equiv) in a microwave oven. The reaction mixture was heated to 140°C and stirred for 3 days. After completion of the reaction, the reaction solution was filtered through potassium fluoride solid wash, and the solid was washed with ethyl acetate. The filtrate was concentrated, and the resulting residue was directly purified by column chromatography with methanol / dichloromethane (0-10%) to obtain compound 196-2 (292 mg, 45.0% yield).

[0413] LCMS: (ESI, m / z): 572.2 [M+H] + .

[0414] Step 3: Synthesis of Compound 196

[0415] Referring to the preparation method of tetrazole in compound 2, compound 196 (78.57 mg, yield 37.75%) was obtained.

[0416] LCMS: (ESI, m / z): 408.2 [M+H] + .

[0417] 1 H NMR(400MHz,CD3OD)δ9.10-9.08(m,1H),7.88-7.86(m,1H),7.73-7.70(m,1H), 6.48(s,1H),6.40(s,1H),4.32(d,J=14.4Hz,1H),3.76(d,J=14.4Hz,1H),2.91 -2.85(m,3H),2.77-2.71(m,2H),2.68-2.61(m,1H),2.60-2.51(m,1H),2.40(d ,J=7.2Hz,2H),1.92-1.85(m,1H),1.09(d,J=6.0Hz,3H),0.92(d,J=6.4Hz,6H).

[0418] Example 55 Preparation of Compound 197

[0419] The preparation method of reference compound 3 was used replace After two steps of reaction, compound 197 (46.29 mg, yield 47.18%) was obtained by column chromatography.

[0420] LCMS: (ESI, m / z): 512.7 [M+H] + .

[0421] 1 H NMR (400MHz, CD3OD) δ9.09–9.07(m,1H),7.84–7.81(m,1H),7.72–7.68(m,1H),6.81(d, J=3.2Hz,2H),4.20(d,J=14.4Hz,1H),3.72(d,J=14.0Hz,1H),3.04–2.99(m,3H),2.94–2 .83(m,4H),2.66–2.61(m,2H),2.52(d,J=7.2Hz,2H),2.46–2.42(m,1H),2.26–2.20(m, 1H),1.94–1.76(m,3H),1.50–1.44(m,2H),0.98(d,J=6.4Hz,3H),0.93(d,J=6.8Hz,6H).

[0422] Example 56 Preparation of Compound 198

[0423] Step 1: Synthesis of Compound 198-1

[0424] To a solution of compound 3-1 (300.0 mg, 0.7 mmol, 1.0 equiv) in tetrahydrofuran (5 mL) at -78°C in a three-necked flask, n-butyllithium (0.9 mL, 1.6 M, 2.0 equiv) was added. The mixture was stirred under nitrogen for 30 minutes, then carbon dioxide gas was introduced. The reaction mixture was stirred at room temperature for 16 hours. After the reaction, the reaction mixture was added with glacially dilute hydrochloric acid solution and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to afford compound 198-1 (60.0 mg, 21.77%) as a yellow solid.

[0425] LCMS: (ESI, m / z): 394.1 [M+H] + .

[0426] Step 2: Synthesis of Compound 198-2

[0427] To a solution of compound 198-1 (60.0 mg, 0.152 mmol, 1.0 equiv) in N,N-dimethylformamide (2 mL) was added N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (87 mg, 0.228 mmol, 1.5 equiv), N,N-diisopropylethylamine (60 mg, 0.457 mmol, 3.0 equiv), and 3,3-difluorocyclobutane-1-amine (17 mg, 0.152 mmol, 0.1 equiv) in a microwave tube. The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was quenched by adding ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to give yellow solid compound 198-2 (40.0 mg, 54.36%).

[0428] LCMS: (ESI, m / z): 483.3 [M+H] + .

[0429] Step 3: Synthesis of Compound 198

[0430] Referring to the preparation method of tetrazole in compound 2, compound 198 (10.74 mg, yield 24.65%) was obtained.

[0431] LCMS: (ESI, m / z): 526.2 [M+H] + .

[0432] 1 H NMR (400MHz, CD3OD) δ9.10-9.08(m,1H),7.84-7.81(m,1H),7.72-7.69(m,1H ),7.15(d,J=6.4Hz,2H),4.27(d,J=14.4Hz,1H),4.08-4.03(m,1H),3.78(d,J =14.4Hz,1H),2.88-2.79(m,5H),2.72-2.70(m,1H),2.63-2.45(m,6H),2.39- 2.34(m,1H),1.98-1.92(m,1H),0.99(d,J=6.0Hz,3H),0.95(d,J=6.4Hz,6H).

[0433] Example 57 Preparation of Compound 199

[0434] Step 1: Synthesis of Compound 199-1

[0435] To a solution of compound 3-1 (200.0 mg, 0.468 mmol, 1.0 equiv) in 1,4-dioxane (5 mL) in a microwave tube, 4-methylpyrazole (77.0 mg, 0.936 mmol, 2.0 equiv), (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (13 mg, 0.094 mmol, 0.2 equiv), cuprous iodide (9.0 mg, 0.047 mmol, 0.1 equiv), and potassium carbonate (194 mg, 1.405 mmol, 3.0 equiv) were added. The reaction mixture was heated to 110°C under nitrogen and stirred for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of ice water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography with methanol / dichloromethane (0-10%) to give yellow solid compound 199-1 (72.0 mg, 35.86%).

[0436] LCMS: (ESI, m / z): 430.3 [M+H] + .

[0437] Step 2: Synthesis of Compound 199

[0438] Referring to the preparation method of tetrazole in compound 2, compound 199 (26.47 mg, yield 19.85%) was obtained.

[0439] LCMS: (ESI, m / z): 473.3 [M+H] + .

[0440] 1 H NMR(400MHz,DMSO-d6)δ9.11–9.10(m,1H),7.68–7.62(m,2H),7.42(s,1H),7.22(s,1H) ,7.12(d,J=1.2Hz,1H),7.05(s,1H),4.02(d,J=14.4Hz,1H),3.59(d,J=14.0Hz,1H),2. 86–2.79(m,2H),2.73–2.69(m,1H),2.55(d,J=7.2Hz,3H),2.47–2.44(m,1H),2.29(s,1 H),2.05–2.00(m,1H),1.96-1.89(m,4H),0.91(d,J=6.4Hz,6H),0.85(d,J=6.0Hz,3H).

[0441] The following compounds were synthesized according to the general preparation method of the present invention and the examples described above:

[0442] Test Example 1: Determine the affinity of the test compound for the AT2 receptor using the AT2R Binding assay

[0443] Experimental procedures:

[0444] 1. Reagent Preparation

[0445] 1.1 Cell line: Tag-lite Angiotensin AT2 labeled Cell;

[0446] 1.2 Experimental buffer: 1X TLB (5 mL 5XTLB + 20 mL HO);

[0447] 2. Activity determination of test compounds

[0448] 2.1 Prepare 1X TLB assay buffer, 4X working solution of the test compound, and 4X Tag-lite angiotensin receptor red agonist (Cisbio, L0007RED).

[0449] 2.2 Freeze-thaw one tube of labeled-AT2R cells in a 37°C water bath until the ice is completely melted (1-2 min). Quickly transfer the thawed cells to 5 mL of 1X TLB, mix gently, and centrifuge at 200g for 5 min.

[0450] 2.3 Discard the supernatant, resuspend the cells in 1 ml of 1X TLB, add 1.7 ml of 1X TLB, mix well, and bring to room temperature for later use;

[0451] 2.4 Add 10 μl of cells to a 384-well plate (Greiner, 784075) and incubate at 200g for 3 s at room temperature. Add 5 μl of 4X compound to each well of the 384-well plate. Add 5 μl of 4X Tag-lite angiotensin receptor red agonist to all test wells.

[0452] 2.5 After the reaction plate was allowed to stand at room temperature (25°C) for 1 hour, centrifuged at 200g, RT, for 60 seconds, and data were collected using the Envision HTRF detector.

[0453] 3. Data Analysis

[0454] 3.1% inhibition calculation:

[0455] %inhibition=100-(Signalcmpd-SignalAve_PC) / (SignalAve_VC-SignalAve_PC)×100.

[0456] 3.2 Calculation of compound IC using GraphPad nonlinear fitting formula 50 :

[0457] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0458] X: log value of compound concentration; Y: % inhibition

[0459] 4. Experimental Results

[0460] The experimental results show that the compound of the present invention has a good binding effect on AT2R.

[0461] Test Example 2: Using the AT1R Binding Assay to Determine the Affinity of the Test Compound for the AT1 Receptor

[0462] Experimental procedures:

[0463] 1. Reagent preparation

[0464] 1) Cell line: Tag-lite Angiotensin AT1 labeled Cell;

[0465] 2) Experimental buffer: 1X TLB (5 mL 5XTLB + 20 mL HO);

[0466] 2. Determination of inhibitory activity of test compounds

[0467] 1) Prepare 1X TLB assay buffer, 4X working solution of the test compound, and 4X Tag-lite angiotensin receptor red agonist (Cisbio, L0007RED).

[0468] 2) Freeze-thaw one tube of labeled-AT1R cells in a 37°C water bath until the ice is completely melted (1-2 min). Quickly transfer the thawed cells to 5 mL of 1X TLB, mix gently, and centrifuge at 200g for 5 min.

[0469] 3) Discard the supernatant, resuspend the cells in 1 ml of 1X TLB, add 1.7 ml of 1X TLB, mix well, and bring to room temperature.

[0470] 4) Add 10 μl of cells to a 384-well plate (Greiner, 784075) and incubate at 200g for 3 s at room temperature. Add 5 μl of 4X compound to the 384-well plate. Add 5 μl of 4X Tag-lite angiotensin receptor red agonist to all test wells.

[0471] 5) After the reaction plate was allowed to stand at room temperature (25°C) for 1 hour, it was centrifuged at 200g, RT, for 60 seconds, and data were collected using an Envision HTRF detector.

[0472] 3. Data Analysis

[0473] 1)Z'factor=1-3*(SDMax+SDMin) / (MeanMax-MeanMin);

[0474] 2)CVMax=(SDMax / MeanMax)*100%;

[0475] 3)CVMin=(SDMin / MeanMin)*100%;

[0476] 4) S / B = Single / Background;

[0477] 5)Calculation Equation for EC 50 / IC 50 Value:

[0478] Y=Bottom+(Top-Bottom) / (1+10^((LogEC 50 -X)*HillSlope))

[0479] 4. Experimental Results

[0480] The experimental results show that the compounds of the present invention have no binding effect on AT1R. (Note: NA: not tested)

[0481] Test Example 3: Study on the pharmacokinetic behavior of the compound of the present invention in mice

[0482] Experimental plan:

[0483] Three healthy male ICR mice (SPF grade, source: Beijing Weitonglihua Laboratory Animal Technology Co., Ltd.), weighing 18-25 g, were intravenously administered with 1 mg / kg of the compound in a 5 ml / kg solution of 5% DMSO, 10% Solutol, and 85% Saline (w / v). The animals were fasted prior to the experiment.

[0484] Three healthy male ICR mice (SPF grade, source: Beijing Weitonglihua Laboratory Animal Technology Co., Ltd.), weighing 18-25 g, were gavaged with 5 mg / kg of the compound in a 10 ml / kg solution of 5% DMSO, 10% Solutol, and 85% Saline (w / v). The animals were fasted prior to the experiment.

[0485] Blood samples were collected from the cheek at 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after intravenous administration and at 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after oral administration. Approximately 0.05 mL of blood was collected for each sample. Blood samples were anticoagulated with sodium heparin and placed on ice. Plasma was separated by centrifugation within 1 hour (centrifugation conditions: 6000g, 3 minutes, 2-8°C). Plasma compound concentrations were determined by liquid chromatography-tandem mass spectrometry. Plasma samples were stored at -80°C until analysis. Pharmacokinetic parameters were calculated using Phoenix WinNonlin 8.2.0 using plasma concentration data at different time points. The experimental results are as follows:

[0486] Reference compound A174: The obtained product was synthesized by referring to the preparation method of WO2023006893A1.

[0487] The experimental results show that the compound of the present invention has a low clearance rate in rodent ICR mice, a high plasma exposure, good oral availability, good pharmacokinetic properties, and is conducive to drug development.

[0488] Test Example 4: Study on the pharmacokinetic behavior of the compound of the present invention in rats

[0489] Experimental drugs: Compounds of the present invention, homemade.

[0490] Experimental plan:

[0491] Three healthy male SD rats (SPF grade, source: Beijing Weitonglihua Experimental Animal Technology Co., Ltd.), weighing 180-250 g, were intravenously injected with 1 mg / kg of the compound in a volume of 2 ml / kg. The compound was prepared using 5% DMSO + 10% Solutol + 85% Saline (w / v). The animals were not fasted before the experiment.

[0492] Three healthy male SD rats (SPF grade, source: Beijing Weitonglihua Experimental Animal Technology Co., Ltd.), weighing 180-250 g, were orally administered with 5 mg / kg of the compound in a volume of 10 ml / kg. The compound was prepared using 5% DMSO + 10% Solutol + 85% Saline (w / v). The animals were not fasted before the experiment.

[0493] Blood samples were collected venously at 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after intravenous administration and at 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after oral administration. Approximately 0.25 mL of blood was collected for each sample. Blood samples were anticoagulated with sodium heparin and placed on ice. Plasma was separated by centrifugation within 1 hour (centrifugation conditions: 6000g, 3 minutes, 2-8°C). Plasma compound concentrations were determined by liquid chromatography-tandem mass spectrometry. Plasma samples were stored at -80°C prior to analysis. Pharmacokinetic parameters were calculated using Phoenix WinNonlin 8.2.0 using plasma drug concentration data at different time points. The experimental results are as follows:

[0494] The experimental results show that the compound of the present invention has a high plasma exposure in rodent SD rats and has good pharmacokinetic properties.

[0495] Test Example 5: Study on the analgesic effect of the compound of the present invention in inhibiting pain in mice in a selective nerve injury model

[0496] 1. Experimental drugs

[0497] Examples 6, 16, 23, 31 and control compound A174.

[0498] A solution of 5% DMSO + 10% Solutol + 85% Saline was used.

[0499] 2. Experimental methods and materials

[0500] 2.1 Experimental animals and housing conditions

[0501] Experimental animals: C57BL / 6 mice, weighing 18-22 g, male, purchased from Beijing Weitonglihua Laboratory Animal Co., Ltd.

[0502] Rearing conditions: 4 animals / cage, 12 / 12 h light / dark cycle, temperature 22-25°C, humidity 40-70%, free access to food and water.

[0503] 2.2 Experimental methods:

[0504] 1.1 Adaptation

[0505] After the experimental animals arrive, they are acclimated and fed for 3-7 days.

[0506] 1.2 Model establishment

[0507] (1) The surgical procedure must be performed aseptically.

[0508] (2) Surgical instruments (scissors, tweezers, scalpels, surgical cotton and sutures) were sterilized before surgery.

[0509] (3) Anesthetize the animal with 50 mg / kg of Zotai 50 + 8 mg / kg of xylazine hydrochloride injection (intraperitoneal injection, 2.5 mL / kg). Squeeze the animal's toes to confirm that the animal is fully anesthetized before surgery; apply ophthalmic ointment to the animal's eyes to prevent the animal's cornea from drying out.

[0510] (4) Shave the hair of the animal's surgical area and disinfect the skin three times with iodine tincture and 70% ethanol; start the surgery after the skin is dry.

[0511] (5) Expose the sciatic nerve and its three distal branches: the tibial nerve, the common peroneal nerve, and the sural nerve. Use ophthalmic scissors to cut the tibial nerve and the common peroneal nerve, while preserving the sural nerve.

[0512] (6) Suture the wound.

[0513] (7) Clean surgical instruments and sterilize them using a hot bead sterilizer.

[0514] (8) After surgery, place the animal on an electric blanket and inject 1 mL of normal saline subcutaneously to prevent dehydration. Once the animal is fully awake (free to move), return it to its cage.

[0515] 1.3 Grouping

[0516] On days 4-6 after modeling, the animals were acclimated to the experimental environment for 30 minutes per day for 3 consecutive days. On day 7 after modeling, the baseline value of mechanical allodynia was measured. Animals that did not show mechanical allodynia (withdrawal threshold greater than 0.6g) were eliminated, and 72 animals with successful modeling (withdrawal threshold less than 0.6g) were selected and divided into 9 groups of 8 animals each.

[0517] 1.4 Administration

[0518] Each group of mice was given a single intraperitoneal or oral dose of 10 mL / kg of the corresponding drug according to the experimental protocol.

[0519] 1.5 Mechanical allodynia test

[0520] ●On the 8th day after model establishment, mechanical allodynia test was performed on the left hind paw of mice at 1, 2 and 4 hours after drug administration.

[0521] ● Place the mouse individually in a plexiglass box with a mesh bottom to ensure that the mouse's feet are accessible for testing. The mouse will be allowed to acclimate for 15 minutes before testing.

[0522] After acclimation, test the center of the sole of the mouse's left hind paw using a test fiber. The test fiber includes eight test intensities: 2.36 (0.02g), 2.44 (0.04g), 2.83 (0.07g), 3.22 (0.16g), 3.61 (0.4g), 3.84 (0.6g), 4.08 (1g), and 4.17 (1.4g). During the test, press the test fiber vertically against the skin and apply force to bend the fiber for 6-8 seconds, with a 5-second interval between each test. During the test, if the animal quickly withdraws its foot, it is recorded as a pain response. If the animal withdraws its foot when the test fiber leaves the animal's skin, it is also recorded as a pain response. If the animal moves or walks, the pain response is not recorded and the test should be repeated.

[0523] ●During the test, first use 3.22 (0.16g). If the animal reacts to pain, use a test fiber of a smaller force in the next test. If the animal does not react to pain, use a test fiber of a larger force in the next test (Chaplan et al. 1994).

[0524] The maximum force of the tested fiber was 4.17 (1.4 g).

[0525] The test results are recorded in the table below, with x being the sign of pain response and o being the sign of no pain response.

[0526] Mechanical allodynia is expressed as the paw withdrawal threshold (PWT) in behavioral tests in mice and is calculated according to the following formula:

[0527] 50% reaction threshold (g) = (10 (Xf+kδ) ) / 10,000

[0528] Xf = Final test fiber value used in the test

[0529] k = table value (Chaplan et al. 1994, page 62)

[0530] δ = mean difference

[0531] 1.6 Key Instruments

[0532] 1.7 Timeline as shown in Figure 1

[0533] 1.8 Experimental test indicators

[0534] 2.3 Data Statistics

[0535] Data were collected using Excel software.

[0536] Data were analyzed using one-way or two-way analysis of variance with Dunnett's multiple comparison test in Prism (Graph Pad Software, Inc.). The area under the analgesic efficacy curve (AUC) was calculated using Prism (Graph Pad Software, Inc.) software. First, select the XY analysis type, enter the data, click Analyze, and select Area under the curve (AUC) in the XY analyses section.

[0537] 3. Results

[0538] In the mouse selective nerve injury model, the area under the analgesic effect of each compound on the pain of mice at 1, 2 and 4 hours after administration was calculated (AUC 0-4h ). The larger the AUC value, the better the analgesic effect.

[0539] Note: P value (compared with the vehicle control group), ***: P < 0.001; **: P < 0.01; ns: no statistically significant difference.

[0540] 4. Conclusion

[0541] The compound of the present invention has a good analgesic effect of inhibiting pain in mice in a selective nerve injury model of mice.

[0542] Test Example 6: Study on the analgesic effect of the compound of the present invention in inhibiting pain in rats in a chronic constriction injury model of sciatic nerve

[0543] 1. Experimental drugs

[0544] Example 35 and reference compound A174.

[0545] A solution of 5% DMSO + 10% Solutol + 85% Saline was used.

[0546] 2. Experimental Materials and Methods

[0547] 2.1 Experimental Materials

[0548] 2.1.1 Experimental animals and housing conditions

[0549] Experimental animals: Sprague-Dawley rats, weighing 80-100 g, male, purchased from Beijing Weitonglihua Experimental Animal Co., Ltd. Housing conditions: 4 rats / cage, 12 / 12 h light / dark cycle, temperature 22-25°C, humidity 40-70%, free access to food and water.

[0550] 2.1.2 Key experimental equipment

[0551] 2.2 Experimental methods:

[0552] 2.2.1 Adaptation

[0553] After the experimental animals arrive, they are acclimated and fed for 3-7 days.

[0554] 2.2.2 Model establishment

[0555] 1) Perform aseptic operation during the surgery.

[0556] 2) Surgical instruments (scissors, forceps, scalpels, surgical cotton, sutures) were sterilized before surgery.

[0557] 3) Anesthetize the animal with Zolta 50 plus Xylazine Hydrochloride Injection (20 mg / kg + 8 mg / kg, intraperitoneally). Squeeze the animal's toes to confirm that the animal is fully anesthetized before surgery. Apply ophthalmic ointment to the animal's eyes to prevent corneal drying.

[0558] 4) Disinfect the surgical area on the sole of the left hind foot three times with iodine and 70% ethanol. Once the skin is dry, begin the surgery.

[0559] 5) The left sciatic nerve was isolated and four loose ligatures were placed approximately 1 mm apart using 4-0 chromic gut approximately 7 mm upstream of the sciatic nerve bifurcation.

[0560] 6) Suture the wound.

[0561] 7) Clean surgical instruments and sterilize them using a hot bead sterilizer.

[0562] 8) After surgery, place the animal on an electric blanket and inject 5 mL of normal saline subcutaneously to prevent dehydration. Once the animal is fully awake (freely moving), return it to its cage.

[0563] 2.2.3 Basic value test grouping:

[0564] 1) Place the animals in the test environment on days 7 to 9 after surgery and acclimate for at least 30 minutes each day.

[0565] 2) On the 10th day after surgery, all animals were tested for mechanical pain baseline (PWT). Model animals that did not show mechanical allodynia (PWT greater than 5g) were eliminated and 24 animals were randomly divided into 3 groups, with 8 animals in each group.

[0566] 2.2.4 Weighing and Dosing

[0567] On the 11th day after surgery, the animals in all groups were weighed and medicated according to the group information.

[0568] 2.2.5 Efficacy test

[0569] Mechanical pain testing was performed on each animal using the test fibers 1 hour and 2 hours after a single administration.

[0570] 2.2.6 Mechanical pain test method:

[0571] 1) Place the rat individually in a plexiglass box with a mesh bottom to ensure the rat's feet are accessible for testing. Allow the rat to acclimate for 15 minutes before testing.

[0572] 2) After acclimation is completed, use the test fiber to test the center of the sole of the left hind paw of the rat. The test fiber includes 8 test strengths: 3.61 (0.4g), 3.84 (0.6g), 4.08 (1g), 4.31 (2g), 4.56 (4g), 4.74 (6g), 4.93 (8g), 5.18 (15g). During the test, the test fiber is pressed vertically to the skin and force is applied to bend the fiber for 6-8 seconds, with an interval of 5 seconds between each test. During the test, the animal's rapid retraction of its foot is recorded as a pain response. The animal's retraction of its foot when the test fiber leaves the animal's skin is also recorded as a pain response. If the animal moves or walks, the pain response is not recorded and the test should be repeated.

[0573] 3) During the test, first use 4.31 (2g). If the animal reacts to pain, the next test uses a test fiber with a lower force. If the animal does not react to pain, the next test uses a test fiber with a higher force. The maximum force of the test fiber is 5.18 (15g).

[0574] 4) The test results are recorded in the table below, with x being the result of a pain response and o being the result of no pain response.

[0575] Mechanical allodynia is expressed as the paw withdrawal threshold (PWT) in the behavioral test of rats and is calculated according to the following formula: 50% response threshold (g) = (10 (Xf+k) ) / 10,000 Xf = Final test fiber value used in the test k = Table value (Chaplan et al. 1994, page 62) δ = Mean difference (0.224)

[0576] 2.2.7 Timeline as shown in Figure 2

[0577] 2.2.8 Experimental test indicators:

[0578] 2.2.9 Data Collection and Analysis:

[0579] Data were collected using Excel software.

[0580] Data were analyzed using Prism (Graph pad software, Inc.) software.

[0581] 3. Experimental Results

[0582] Note: P value (compared with the vehicle control group), ***: P < 0.001; **: P < 0.01.

[0583] 4. Conclusion

[0584] The compound of the present invention has a good analgesic effect of inhibiting pain in rats in a chronic constriction injury model of the sciatic nerve.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, in: M1 and M2 are each independently selected from C or N; L1 is a bond or -CH2-; L2 is a bond, -CH2-, -CF2-, -C(O)-, -C(O)NH-, -NHC(O)-, -NH-, -O- or -S-, wherein -CH2- and -NH- are optionally replaced by one or two R L2 Replacement, R L2 Each independently selected from halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl, C 1- 3 haloalkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; L is selected from L3 or ring A; L3 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy or C 1-6 hydroxyalkyl; Ring A is a monocyclic or bicyclic ring, specifically selected from C 3-10 cycloalkyl, 3-10 membered heterocyclic group, C 6-14 Aryl, 5-10 membered heteroaryl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl, or 5-6 membered heterocyclyl and aryl; R 1 Each independently selected from halogen, hydroxyl, amino, amide, C 1-3 Alkylamino, di(C 1-3 alkyl)amino, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; Or, any two R 1 Connect to form a 3-8 membered heterocyclic group or C 3-8 Cycloalkyl; R 2 Selected from amino, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl, -OR 2.1 or -NHR 2.1 ; the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl, optionally further substituted by one or more R 2.1 replace; R 2.1 Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, the C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl and 3-6 membered heterocyclic groups, optionally substituted by one or more R 2.1.1 replace; R 2.1.1 Selected from halogen, amino, cyano, hydroxyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; R 3 Each independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl or C 1-6 haloalkoxy; R 4 is selected from 5-6 membered heteroaryl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl, 5-6 membered heterocyclylphenyl or 5-6 membered heterocyclyl, the 5-6 membered heteroaryl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl, 5-6 membered heterocyclylphenyl and 5-6 membered heterocyclyl, optionally substituted by one or more R 4.1 replace; R 4.1 Selected from halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; x and y are each independently selected from 0, 1, 2, 3 or 4.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that The L1 is selected from -CH2-; or, the R 4 is selected from 5-6 membered heteroaryl, 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroarylphenyl, 5-6 membered heterocyclylphenyl, 5-6 membered heterocyclyl and 5-6 membered heteroaryl or 6 membered heterocyclyl; preferably, the R 4 is selected from thiazolyl, pyrazolyl, pyridazinyl, imidazolyl, pyridinyl, isoxazolyl, triazolyl, tetrazolyl, oxazolyl, pyrazinyl, pyrimidinyl, oxadiazolyl, pyridonyl, pyridazinonyl, pyrimidonyl, pyrazinonyl, imidazopyridinyl, imidazophenyl, thiazophenyl, oxazolophenyl, pyrimidonphenyl, pyrimidonopyridinyl, pyrazolophenyl, pyrimidonphenyl, pyrazinonphenyl, pyrimidonothiazolyl, pyrimidonopyrazolyl or oxazolopyridinyl; more preferably, said R 4 is selected from thiazolyl, pyridazinyl, thiazolophenyl or pyrazinyl.

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, characterized in that The compound of formula (I) is further represented by formula (II):

4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, characterized in that The ring A is selected from 5-6 membered heteroaryl, 5 membered heteroarylphenyl, 5 membered heterocyclylphenyl, 5 membered heterocyclyl and 6 membered heteroaryl, 6 membered heterocyclyl and 5 membered heteroaryl, C 6-10 Aryl, C 3-8 cycloalkyl or 3-8 membered heterocyclic group; preferably, the ring A is selected from pyridazinyl, pyrazolyl, pyridyl, phenyl, pyrrolyl, imidazolyl, triazolyl, tetrazolyl, furyl, thienyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyranyl, pyrrolidinyl, pyridonyl, morpholinyl, piperidinyl, oxirane, aziridine, oxetanyl, azetidinyl, piperazinyl, pyrrolophenyl, pyrrolidinylphenyl, diazaspiro[3.3]heptanyl or oxazaspiro[3.3]heptanyl.

5. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, characterized in that: The R 1 Each independently selected from halogen, hydroxyl, amino, amide, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-4 Cycloalkyl or 3-4 membered heterocyclic group; preferably, the R 1 Each is independently selected from fluorine, chlorine, hydroxy, amino, amido, cyano, methyl, ethyl, propyl, methoxy, ethoxy, propoxy, trifluoromethyl, difluoromethyl, trifluoroethyl, hydroxymethyl, hydroxyethyl, cyclopropyl, cyclobutyl, oxirane, aziridine, oxetanyl or azetidinyl.

6. The compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, characterized in that: The L2 is selected from -NH-, -CH2-, -CF2-, -C(O)NH-, -NHC(O)-, -O- or -S-.

7. The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, characterized in that: The R 2 Selected from amino, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, -OR 2.1 or -NHR 2.1 Preferably, the R 2 Selected from amino, methyl, ethyl, propyl, n-butyl, isobutyl, cyclopropyl, piperidinyl, morpholinyl, piperazinyl, -OR 2.1 or -NHR 2.1 ; R 2.1 Selected from C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-4 Cycloalkyl, 3-4 membered heterocyclic group, the C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkyl and 3-6 membered heterocyclic groups, optionally substituted by one or more R 2.1.1 substituted; preferably, R 2.1 is selected from methyl, ethyl, propyl, n-butyl, isobutyl, methoxy, ethoxy, propoxy, cyclopropyl, cyclobutyl, cyclopentyl, oxirane, aziridine, oxetanyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, trifluoromethyl, difluoromethyl or difluoroethyl; The R 2.1.1 Selected from halogen, amino, cyano, hydroxyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group; preferably, the R 2.1.1 is selected from fluorine, chlorine, methyl, ethyl, propyl, n-butyl, isobutyl, methoxy, ethoxy, propoxy, cyclopropyl, cyclobutyl, cyclopentyl, oxirane, aziridine, oxetanyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, trifluoromethyl, difluoromethyl or difluoroethyl; Or, R 3 Selected from hydrogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl or C 1-4 preferably, R 3 is selected from hydrogen, methyl, ethyl, propyl, methoxy, ethoxy, propoxy, trifluoromethyl or difluoromethyl.

8. The compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, characterized in that The compound is selected from the compounds shown in Table 1.

9. A pharmaceutical composition comprising: (1) The compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, and (2) Pharmaceutically acceptable excipients.

10. Use of the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 9, in the preparation of a medicament for preventing and / or treating pain; preferably, the pain is neuropathic pain.

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