RU2800296C1 - Oxaspiro derivative substituted at azabicyclic ring, a method of its production and its medical use - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to a compound represented in formula (I) or a pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers thereof, wherein ring A is pyridin-2-yl; (R₀)_n means that the hydrogen atoms in ring A are replaced by n R₀ groups, where n is 0; Y₁ is N; Y₂ is a bond; Y₁ is directly connected to N; Y₃ is CR₃; Y₄ is CR₄; R₃ and R₄ are independently hydrogen or halogen; W is CH₂; R_a, R_b, R_c and R_d are defined as follows: (i) each of R_a and R_b independently represents hydrogen or C_1-8alkyl; or R_a and R_b together with their associated carbon atom form a three to six membered saturated monocyclic ring; R_c is hydrogen, halogen, C_1-8alkyl, C_1-8alkoxy, or halogenated C_1-8alkyl; or R_d is hydrogen or C_1-8alkyl; or (ii) each of R_a and R_b independently represents hydrogen; or R_c is linked to R_d to form a three to six membered saturated monocyclic ring or a four to six membered saturated monoheterocyclic ring, wherein the heteroatom in the four to six membered saturated monoheterocyclic ring is O or N; and m is either 0 or 1. The invention also relates to intermediate compounds of formula (I-2b).

EFFECT: new compounds have been obtained that can be used in medicine as therapeutic agents for the treatment and prevention of pain and diseases associated with pain.

14 cl, 2 tbl, 23 ex

Description

FIELD OF THE INVENTION

[0001] The present disclosure relates to the class of oxaspirocyclic derivatives substituted on the azabicyclic ring, methods for their preparation, pharmaceutical compositions containing the derivatives, and their use as therapeutic agents, in particular as MOR receptor agonists, and their use in the preparation of drugs for the treatment and prevention of pain and diseases associated with pain.

BACKGROUND OF THE INVENTION

[0002] Opioid receptors are an important class of G protein-coupled receptors (GPCRs) that bind endogenous opioid peptides and opioid drugs. Endogenous opioid peptides are opioid-like active substances that are naturally produced in mammals, and known endogenous opioid peptides are broadly classified into enkephalins, endorphins, dynorphins, and neoendorphins. The corresponding opioid receptors, namely μ (MOR) receptor, δ (DOR) receptor, κ (KOR) receptor, etc., are present in the central nervous system. Studies have established that the analgesic effect of endogenous opioid peptides mainly depends on the level of expression of opioid receptors, targets that are affected by the analgesic effects of opioid drugs, as well as endogenous opioid peptides.

[0003] Current research suggests that GPCRs mediate and regulate physiological functions primarily through two signaling pathways: the G-protein signaling pathway and the β-arrestin signaling pathway. After a conventional GPCR agonist binds to the receptor, the G-protein signaling pathway is activated, which includes second messenger systems such as calcium ions, adenyl cyclase (AC), and mitogen-activated protein kinases (MAPKs), while the arrestin-associated β-ligand mainly activates the β-arrestin signaling pathway. By mediating GPCR responses, β-arrestin mainly acts as follows: 1) as a negative regulator, it acts with G protein-coupled receptor kinase (GRK) to desensitize GPCR and stop G protein signaling; 2) as a scaffold protein, it recruits endocytic proteins and induces GPCR endocytosis; and 3) as an adapter protein, it forms a complex with downstream GPCR signaling molecules to activate signal transduction molecules such as MAPK, protein tyrosine kinase Src and Akt in a G-protein independent manner. Differences in ligand-stimulated G-protein signaling and/or β-arrestin signaling ultimately determine the ligand-specific cellular biological effects of GPCR.

[0004] MOR is the target of endogenous enkephalins and opioid analgesics such as morphine. Early studies have shown that endogenous enkephalins and opioid drugs such as etorphine can activate G proteins and trigger receptor endocytosis, but morphine does not trigger receptor endocytosis at all because morphine is too weak to combat MOR phosphorylation and can only capture trace amounts of β-arrestin on the cell membrane (Zhang et al., Proc Natl Acad Sci USA, 19 98, 95(12): 7157-7162). Such ligands carry out their physiological functions exclusively through the G-protein signaling pathway, and not through the β-arrestin pathway. Studies have shown that the analgesic effect mediated by G-protein signaling is stronger and lasts longer after morphine injection in β-arrestin-2 knockout mice (Bohn et al., Science, 1999). It would be expected that if such ligands are highly negatively associated with β-arrestin and can even avoid β-arrestin-mediated receptor desensitization, the ligands could lead to prolonged G-protein signaling and stronger analgesic effects.

[0005] Currently, MOR agonists are disclosed in patent applications WO2017106547, WO2017063509, WO2012129495, WO2017106306, etc.

[0006] Prolonged use of opioid drugs causes tolerance as well as side effects such as respiratory depression and constipation, which have been shown to be closely related to the functions of β-arrestin. In order to reduce the side effects of opioid drugs, drugs can be designed based on β-arrestin-based negative MOR ligands to reduce β-arrestin-mediated side effects and enhance therapeutic effects.

SUMMARY OF THE INVENTION

[0007] The purpose of the present disclosure is to provide a compound that has a novel structure and can be used as a MOR agonist.

[0008] In a first aspect, the present disclosure provides a compound represented in formula (I), or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate or prodrug thereof:

(I)

[0009] where

[0010] Ring A is a C _6-10 aromatic ring or a five- or six-membered monocyclic heteroaryl ring;

[0011] (R₀)_n means that the hydrogen atoms in ring A are replaced by n groups R₀where n is 0, 1, 2, 3 or 4 and n is R₀ are the same or different and are each independently hydrogen, cyano, acetyl, hydroxyl, hydroxymethyl, hydroxyethyl, carboxyl, halogenated C_1-8 alkyl (preferably halogenated C_1-4 alkyl), halogen (preferably F or Cl), nitro, C_6-10 aryl (preferably phenyl), five- or six-membered monocyclic heteroaryl, C_1-8 alkyl (preferably C_1-4 alkyl), C_1-8 alkoxy (preferably C_1-4 alkoxy), C_3-6 cycloalkyl, C_3-6 cycloalkoxy, C_2-4 alkenyl, C_2-4 alkynyl, NR_elevenR₁₂, -CONR_elevenR₁₂, -C(O)O(C_1-8 alkyl) (preferably -C(O)O(C_1-4 alkyl)), -OC(O)(C_1-8 alkyl) (preferably -OC(O)(C_1-4 alkyl)), -SO₂(C_1-8 alkyl) (preferably -SO₂(C_1-4 alkyl)), -SO₂(C_6-10 aryl) (preferably -SO₂(C₆ aryl), such as -SO₂-phenyl), -CO(C_6-10 aryl) (preferably -CO(C₆ aryl), such as -CO-phenyl), a four to six membered saturated monoheterocyclic ring or a three to six membered saturated monocyclic ring in which each C_6-10 aryl, five or six membered monocyclic heteroaryl, four to six membered saturated monoheterocyclic ring and three to six membered saturated monocyclic ring is unsubstituted or substituted with one, two or three substituent groups selected from the group consisting of acetyl, hydroxyl, cyano, halogen, C_1-3 alkyl, C_1-3 alkoxy, C_3-6 cycloalkyl and NR_elevenR₁₂;

[0012] Y ₁ is CR ₁ or N;

[0013] Y ₂ is a bond, NR ₂ , C=O or SO ₂ ;

[0014] Y ₃ is CR ₃ or N;

[0015] Y ₄ is CR ₄ or N;

[0016] if Y ₂ is a bond, Y ₁ is directly linked to N;

[0017] each of R₁, R₃ and R₄ is independently hydrogen, cyano, hydroxyl, hydroxymethyl, cyanomethyl, halogen, C_1-8 alkoxy (preferably C_1-4 alkoxy), C_1-8 alkyl (preferably C_1-4 alkyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂ or -NHSO₂(C_3-6 cycloalkyl);

[0018] R ₂ is hydrogen, C _1-8 alkyl (preferably C _1-4 alkyl), -CO(C _1-8 alkyl) (preferably -CO(C _1-4 alkyl)), -CONR ₁₁ R ₁₂ or -SO ₂ (C _1-8 alkyl) (preferably -SO ₂ (C _1-4 alkyl));

[0019] W is CR _w1 R _w2 , NR _w3 , O or C=O,

[0020] each of R _w1 and R _w2 is independently hydrogen, halogen (preferably F or Cl), C _1-8 alkyl (preferably C _1-4 alkyl), halogenated C _1-8 alkyl (preferably halogenated C _1-4 alkyl), or halogenated C _1-8 alkoxy (preferably halogenated C _1-4 alkoxy); или R _w1 и R _w2 вместе со связанным с ними атомом углерода образуют четырех-шестичленное насыщенное моногетероциклическое кольцо или трех-шестичленное насыщенное моноциклическое кольцо, где каждое четырех-шестичленное насыщенное моногетероциклическое кольцо и трех-шестичленное насыщенное моноциклическое кольцо не замещено или замещено одной, двумя или тремя замещающими группами, выбранными из группы, состоящей из циано, гидроксила, гидроксиметила, цианометила, галогена, C _1-8 алкоксила (предпочтительно C _1-4 алкоксила), C _1-8 алкила (предпочтительно C _1-4 алкила), -CO(C _1-8 алкила) (предпочтительно -CO(C _1-4 алкила)), -CONR ₁₁ R ₁₂ , NR ₁₁ R ₁₂ , -NHCO(C _1-8 алкила) (предпочтительно -NHCO(C _1-4 алкила)), -NHCONR ₁₁ R ₁₂ , -NHSO ₂ (C _1-8 алкила) (предпочтительно -NHSO ₂ (C _1-4 алкила)), -NHSO ₂ NR ₁₁ R ₁₂ и -NHSO ₂ (C _3-6 циклоалкила);

[0021] R _w3 is hydrogen, C _1-8 alkyl (preferably C _1-4 alkyl) or -CO(C _1-8 alkyl) (preferably -CO(C _1-4 alkyl));

[0022] R _a , R _b , R _c and R _d are defined as follows:

[0023] (I) Each of R _A and R _B is independently of hydrogen, halogen (preferably F or CL), C _1-8 alkyl (preferably C _1-4 alkyl, and more preferably methyl, ethyl, N-spray or isopropil), halogened with _1-8 alkyl (preferably halogenic with _1-4 acills) or halogenic Clogenized C _1-8 alcoxyls) (preferably halogenic with _1-4 alcoxil); или R _a и R _b вместе со связанным с ними атомом углерода образуют четырех-шестичленное насыщенное моногетероциклическое кольцо или трех-шестичленное насыщенное моноциклическое кольцо (предпочтительно трех-пятичленное насыщенное моноциклическое кольцо, а более предпочтительно циклопропильное кольцо, циклобутильное кольцо или циклопентильное кольцо), где каждое четырех-шестичленное насыщенное моногетероциклическое кольцо и трех-шестичленное насыщенное моноциклическое кольцо не замещено или замещено одной, двумя или тремя замещающими группами, выбранными из группы, состоящей из циано, гидроксила, гидроксиметила, цианометила, галогена, C _1-8 алкоксила (предпочтительно C _1-4 алкоксила), C _1-8 алкила (предпочтительно C _1-4 алкила), -CO(C _1-8 алкила) (предпочтительно -CO(C _1-4 алкила)), -CONR ₁₁ R ₁₂ , NR ₁₁ R ₁₂ , -NHCO(C _1-8 алкила) (предпочтительно -NHCO(C _1-4 алкила)), -NHCONR ₁₁ R ₁₂ , -NHSO ₂ (C _1-8 алкила) (предпочтительно -NHSO ₂ (C _1-4 алкила)), -NHSO ₂ NR ₁₁ R ₁₂ и -NHSO ₂ (C _3-6 циклоалкила);

[0024] R_c is hydrogen, halogen (preferably F or Cl), C_1-8 alkyl (preferably C_1-4 alkyl, and more preferably methyl), C_2-10 alkenyl (preferably C_2-6 alkenyl, and more preferably C_2-4 alkenyl), C_2-10 alkynyl (preferably C_2-6 alkynyl, and more preferably C_2-4 alkynyl), C_1-8 alkoxy (preferably C_1-4 alkoxy, and more preferably methoxy), halogenated C_1-8 alkyl (preferably halogenated C_1-4 alkyl, and more preferably trifluoromethyl), halogenated C_1-8 alkoxy (preferably halogenated C_1-4 alkoxyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂, -NHSO₂(C_3-6 cycloalkyl), -SO₂(C_1-8 alkyl) (preferably -SO₂(C_1-4 alkyl)), -SO₂NR_elevenR₁₂, four to six membered saturated monoheterocyclic ring, C_6-10 aryl or a five- or six-membered monocyclic heteroaryl, where each four-to-six-membered saturated monoheterocyclic ring, C_6-10 aryl and five- or six-membered monocyclic heteroaryl are unsubstituted or substituted with one, two or three substituent groups selected from the group consisting of cyano, hydroxyl, hydroxymethyl, cyanomethyl, halogen, C_1-8 alkoxy (preferably C_1-4 alkoxy), C_1-8 alkyl (preferably C_1-4 alkyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂ and -NHSO₂(C_3-6 cycloalkyl);

[0026] R _d is hydrogen or C _1-8 alkyl (preferably C _1-4 alkyl); or

[0026] (ii) R _a and R _b are each independently hydrogen, halogen (preferably F or Cl), C _1-8 alkyl (preferably C _1-4 alkyl), halogenated C _1-8 alkyl (preferably halogenated C _1-4 alkyl), or halogenated C _1-8 alkoxy (preferably halogenated C _1-4 alkoxy);

[0027] R _c is linked to R _d to form a three to six membered saturated monocyclic ring or a four to six membered saturated monoheterocyclic ring;

[0028] R ₁₁ and R ₁₂ are each independently hydrogen, C _1-8 alkyl (preferably C _1-4 alkyl), halogenated C _1-8 alkyl (preferably halogenated C _1-4 alkyl), C _3-6 cycloalkyl, or a four to six membered saturated monoheterocyclic ring; or R ₁₁ and R ₁₂ together with their associated nitrogen atom form a four to six membered saturated monoheterocyclic ring in which the four to six membered saturated monoheterocyclic ring is unsubstituted or substituted with one, two or three C _1-4 alkyl groups;

[0029] m is 0 or 1.

[0030] In another preferred embodiment, Ring A is a five- or six-membered monoheterocyclic ring which is selected from the group consisting of thiophene, furan, thiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,5-triazole, 1,3,4- triazole, tetrazole, isoxazole, oxadiazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, thiadiazole, pyridine, pyridazine, pyrimidine and pyrazine.

[0031] In another preferred embodiment, Ring A is a benzene ring or a pyridine ring.

[0032] In another preferred embodiment, W is CR _w1 R _w2 .

[0033] In another preferred embodiment, W is CH ₂ .

[0034] In another preferred embodiment, W is NR _w3 or O.

[0035] In another preferred embodiment, W is C=O.

[0036] In another preferred embodiment, R _w1 and R _w2 together with their associated carbon atom form a four to six membered saturated monoheterocyclic ring which is selected from the group consisting of azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, and tetrahydropyran .

[0037] In another preferred embodiment, R _w1 and R _w2 together with their associated carbon atom form a three to six membered saturated monocyclic ring which is selected from the group consisting of cyclopropyl ring, cyclobutyl ring, cyclopentyl ring and cyclohexyl ring.

[0038] In another preferred embodiment, represents .

[0039] In another preferred embodiment, R _c is linked to R _d to form a three to six membered saturated monocyclic ring which is selected from the group consisting of a cyclopropyl ring, a cyclobutyl ring, a cyclopentyl ring, and a cyclohexyl ring.

[0040] In another preferred embodiment, R _c is linked to R _d to form a four to six membered saturated monoheterocyclic ring which is selected from the group consisting of azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, and tetrahydropyran.

[0041] In a second aspect, the present disclosure provides a compound represented in formula (II), or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate or prodrug thereof:

(II)

[0042] where

[0043] Z is CR ₀₁ or N;

[0044] R ₀₁ is hydrogen, cyano, acetyl, hydroxyl, hydroxymethyl, hydroxyethyl, carboxyl, halogenated C _1-8 alkyl (preferably halogenated C _1-4 alkyl), halogen (preferably F or Cl), C _1-8 alkyl (preferably C _1-4 alkyl), C _1-8 alkoxy (preferably C _1-4 alkoxy), C _3-6 cycloalkyl or NR ₁₁ R ₁₂ ;

[0045] (R₀)_n means that the hydrogen atoms in the six-membered ring are substituted by n R groups₀where n is 0, 1, 2, 3 or 4 and n is R₀ are the same or different and are each independently hydrogen, cyano, acetyl, hydroxyl, hydroxymethyl, hydroxyethyl, carboxyl, halogenated C_1-8 alkyl (preferably halogenated C_1-4 alkyl), halogen (preferably F or Cl), nitro, C_6-10 aryl (preferably phenyl), five- or six-membered monocyclic heteroaryl, C_1-8 alkyl (preferably C_1-4 alkyl), C_1-8 alkoxy (preferably C_1-4 alkoxy), C_3-6 cycloalkyl, C_3-6 cycloalkoxy, C_2-4 alkenyl, C_2-4 alkynyl, NR_elevenR₁₂, -CONR_elevenR₁₂, -C(O)O(C_1-8 alkyl) (preferably -C(O)O(C_1-4 alkyl)), -OC(O)(C_1-8 alkyl) (preferably -OC(O)(C_1-4 alkyl)), -SO₂(C_1-8 alkyl) (preferably -SO₂(C_1-4 alkyl)), -SO₂(C_6-10 aryl) (preferably -SO₂C₆ aryl such as -SO₂-phenyl), -CO(C_6-10 aryl) (preferably -COC₆ aryl such as -CO-phenyl), a four to six membered saturated monoheterocyclic ring, or a three to six membered saturated monocyclic ring, where each C_6-10 aryl, five or six membered monocyclic heteroaryl, four to six membered saturated monoheterocyclic ring and four to six membered saturated monocyclic ring is unsubstituted or substituted with one, two or three substituent groups selected from the group consisting of acetyl, hydroxyl, cyano, halogen, C_1-3 alkyl, C_1-3 alkoxy, C_3-6 cycloalkyl and NR_elevenR₁₂;

[0046] Y ₁ is CR ₁ or N;

[0047] Y ₂ is a bond, NR ₂ , C=O or SO ₂ ;

[0048] Y ₃ is CR ₃ or N;

[0049] Y ₄ is CR ₄ or N;

[0050] if Y ₂ is a bond, Y ₁ is directly linked to N;

[0051] each of R₁, R₃ and R₄ is independently hydrogen, cyano, hydroxyl, hydroxymethyl, cyanomethyl, halogen, C_1-8 alkoxy (preferably C_1-4 alkoxy), C_1-8 alkyl (preferably C_1-4 alkyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂ or -NHSO₂(C_3-6 cycloalkyl);

[0052] R ₂ is hydrogen, C _1-8 alkyl (preferably C _1-4 alkyl), -CO(C _1-8 alkyl) (preferably -CO(C _1-4 alkyl)), -CONR ₁₁ R ₁₂ or -SO ₂ (C _1-8 alkyl) (preferably -SO ₂ (C _1-4 alkyl));

[0053] each of R _a and R _b is independently hydrogen, halogen (preferably F or Cl), C _1-8 alkyl (preferably C _1-4 alkyl), halogenated C _1-8 alkyl (preferably halogenated C _1-4 alkyl), or halogenated C _1-8 alkoxy (preferably halogenated C _1-4 alkoxy); или R _a и R _b вместе со связанным с ними атомом углерода образуют четырех-шестичленное насыщенное моногетероциклическое кольцо или трех-шестичленное насыщенное моноциклическое кольцо, где каждое четырех-шестичленное насыщенное моногетероциклическое кольцо и трех-шестичленное насыщенное моноциклическое кольцо не замещено или замещено одной, двумя или тремя замещающими группами, выбранными из группы, состоящей из циано, гидроксила, гидроксиметила, цианометила, галогена, C _1-8 алкоксила (предпочтительно C _1-4 алкоксила), C _1-8 алкила (предпочтительно C _1-4 алкила), -CO(C _1-8 алкила) (предпочтительно -CO(C _1-4 алкила)), -CONR ₁₁ R ₁₂ , NR ₁₁ R ₁₂ , -NHCO(C _1-8 алкила) (предпочтительно -NHCO(C _1-4 алкила)), -NHCONR ₁₁ R ₁₂ , -NHSO ₂ (C _1-8 алкила) (предпочтительно -NHSO ₂ (C _1-4 алкила)), -NHSO ₂ NR ₁₁ R ₁₂ и -NHSO ₂ (C _3-6 циклоалкила);

[0054] R_c is hydrogen, halogen (preferably F or Cl), C_1-8 alkyl (preferably C_1-4 alkyl), C_2-10 alkenyl (preferably C_2-6 alkenyl, and more preferably C_2-4 alkenyl), C_2-10 alkynyl (preferably C_2-6 alkynyl, and more preferably C_2-4 alkynyl), C_1-8 alkoxy (preferably C_1-4 alkoxy), halogenated C_1-8 alkyl (preferably halogenated C_1-4 alkyl), halogenated C_1-8 alkoxy (preferably halogenated C_1-4 alkoxyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂, -NHSO₂(C_3-6 cycloalkyl), -SO₂(C_1-8 alkyl) (preferably -SO₂(C_1-4 alkyl)), -SO₂NR_elevenR₁₂, four to six membered saturated monoheterocyclic ring, C_6-10 aryl or a five- or six-membered monocyclic heteroaryl, where each four-to-six-membered saturated monoheterocyclic ring, C_6-10 aryl and five- or six-membered monocyclic heteroaryl are unsubstituted or substituted with one, two or three substituent groups selected from the group consisting of cyano, hydroxyl, hydroxymethyl, cyanomethyl, halogen, C_1-8 alkoxy (preferably C_1-4 alkoxy), C_1-8 alkyl (preferably C_1-4 alkyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂ and -NHSO₂(C_3-6 cycloalkyl);

[0055] R ₁₁ and R ₁₂ are each independently hydrogen, C _1-8 alkyl (preferably C _1-4 alkyl), halogenated C _1-8 alkyl (preferably halogenated C _1-4 alkyl), C _3-6 cycloalkyl, or a four to six membered saturated monoheterocyclic ring; or R ₁₁ and R ₁₂ together with their associated nitrogen atom form a four to six membered saturated monoheterocyclic ring, where the four to six membered saturated monoheterocyclic ring is unsubstituted or substituted with one, two or three C _1-4 alkyl groups; And

[0056] m is 0 or 1.

[0057] In another preferred embodiment, Y ₁ is N or CH.

[0058] In another preferred embodiment, Y ₂ is a bond or C=O.

[0059] In another preferred embodiment, Y ₃ is CR ₃ .

[0060] In another preferred embodiment, Y ₃ is CH or N.

[0061] In another preferred embodiment, Y ₄ is CR ₄ .

[0062] In another preferred embodiment, Y ₄ is CH or N.

[0063] In another preferred embodiment, Y ₁ is N; Y ₂ is a bond; Y ₃ is CR ₃ or N; Y ₄ is CR ₄ or N; and R ₃ and R ₄ are as defined above.

[0064] In another preferred embodiment, Y ₁ is N; Y ₂ is a bond; Y ₃ is CR ₃ ; Y ₄ is CR ₄ ; and R ₃ and R ₄ are as defined above.

[0065] In another preferred embodiment, Y ₁ is N; Y ₂ is a bond; Y ₃ is CH; and Y ₄ is CH.

[0066] In another preferred embodiment, Y ₁ is N; Y ₂ is C=O; Y ₃ is CR ₃ or N; Y ₄ is CR ₄ or N; and R ₃ and R ₄ are as defined above.

[0067] In another preferred embodiment, Y ₁ is CR ₁ ; Y ₂ is a bond; Y ₃ is CR ₃ or N; Y ₄ is CR ₄ or N; and R ₁ , R ₃ and R ₄ are as defined above.

[0068] In another preferred embodiment, m is 1.

[0069] In another preferred embodiment, Z is N.

[0070] In another preferred embodiment, Z is CR ₀₁ ; and R ₀₁ is hydrogen, cyano, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, halogenated C _1-3 alkyl, F, Cl, C _1-3 alkyl, C _1-3 alkoxy, or C _3-6 cycloalkyl.

[0071] In another preferred embodiment, Z is CH.

[0072] In another preferred embodiment, the compound represented by formula (II) has the structure represented by formula (III):

.

[0073] In another preferred embodiment, the five- or six-membered monocyclic heteroaryl linked to R ₀ is selected from thiophene, furan, thiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,5-triazole, 1,3,4-triazole , tetrazole, isoxazole, oxadiazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, thiadiazole, pyridine, pyridazine, pyrimidine or pyrazine.

[0074] In another preferred embodiment, the four to six membered saturated monoheterocyclic ring linked to R ₀ is selected from the group consisting of azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, and tetrahydropyran.

[0075] In another preferred embodiment, R _a and R _b together with their associated carbon atom form a four to six membered saturated monoheterocyclic ring which is selected from the group consisting of azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide and tetrahydropyran.

[0076] In another preferred embodiment, R _a and R _b together with their associated carbon atom form a three to six membered saturated monocyclic ring which is selected from a cyclopropyl ring, a cyclobutyl ring, a cyclopentyl ring, and a cyclohexyl ring.

[0077] In another preferred embodiment, the five or six membered monocyclic heteroaryl linked to R _c is selected from thiophene, furan, thiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,5-triazole, 1,3,4-triazole, tetrazole, isoxazole, oxadiazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, thiadiazole, pyridine, pyridazine, pyrimidine or pyrazine.

[0078] In another preferred embodiment, the four to six membered saturated monoheterocyclic ring linked to R _c is selected from the group consisting of azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, and tetrahydropyran.

[0079] In another preferred embodiment, the four to six membered saturated monoheterocyclic ring linked to R ₁₁ and R ₁₂ is selected from the group consisting of azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, and tetrahydropyran.

[0080] In another preferred embodiment, R ₁₁ and R ₁₂ together with their associated nitrogen atom form a four to six membered saturated monoheterocyclic ring which is selected from the group consisting of azetidine, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine and thiomorpholine-1,1-dioxide.

[0081] In another preferred embodiment, each of R _a and R _b independently represents hydrogen or C _1-3 alkyl (preferably methyl); or R _a and R _b together with their associated carbon atom form a three to six membered saturated monocyclic ring (preferably a cyclopropyl ring, cyclobutyl ring or cyclopentyl ring).

[0082] In another preferred embodiment, R _c is hydrogen, halogen (preferably F), C _1-3 alkyl (preferably methyl, ethyl, or n-propyl), C _1-3 alkoxy (preferably methoxy), or halogenated C _1-3 alkyl (preferably trifluoromethyl).

[0083] In another preferred embodiment, n is 0.

[0084] In another preferred embodiment, the four to six membered saturated monoheterocyclic ring is selected from the following structures:

[0085]

and the hydrogen atoms in the four to six membered saturated monoheterocyclic ring are optionally substituted with one, two or three substituent groups each independently selected from the group consisting of cyano, hydroxyl, hydroxymethyl, cyanomethyl, halogen, C_1-8 alkoxy (preferably C_1-4 alkoxy), C_1-8 alkyl (preferably C_1-4 alkyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂ and -NHSO₂(C_3-6 cycloalkyl).

[0086] In another preferred embodiment, each of the five or six membered monocyclic heteroaryl linked to the A ring, the five or six membered monocyclic heteroaryl linked to R ₀ , and the five or six membered monocyclic heteroaryl linked to R _c is independently selected from the following structures:

and the five or six membered monocyclic heteroaryl is optionally substituted with one, two or three substituent groups each independently selected from the group consisting of cyano, hydroxyl, hydroxymethyl, cyanomethyl, halogen, C_1-8 alkoxy (preferably C_1-4 alkoxy), C_1-8 alkyl (preferably C_1-4 alkyl), -CO(C_1-8 alkyl) (preferably -CO(C_1-4 alkyl)), -CONR_elevenR₁₂, NR_elevenR₁₂, -NHCO(C_1-8 alkyl) (preferably -NHCO(C_1-4 alkyl)), -NHCONR_elevenR₁₂, -NHSO₂(C_1-8 alkyl) (preferably -NHSO₂(C_1-4 alkyl)), -NHSO₂NR_elevenR₁₂ and -NHSO₂(C_3-6 cycloalkyl).

[0087] In another preferred embodiment, the compound is selected from the group consisting of:

.

[0088] In a third aspect, the present disclosure provides a pharmaceutical composition that includes a compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate, or prodrug thereof, according to the first or second aspect of the present disclosure; and a pharmaceutically acceptable carrier.

[0089] In a fourth aspect, the present disclosure provides the use of a compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate, or prodrug thereof, according to the first or second aspect of the present disclosure, or a pharmaceutical composition according to the third aspect of the present disclosure, in the manufacture of a medicament for the prevention and/or treatment of a disease mediated by an MOR receptor agonist.

[0090] In a fifth aspect, the present disclosure provides the use of a compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate, or prodrug of the first or second aspect of the present disclosure, or a pharmaceutical composition of the third aspect of the present disclosure, in the manufacture of a medicament to activate or antagonize MOR receptors.

[0091] In a sixth aspect, the present disclosure provides the use of a compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate, or prodrug thereof, according to the first or second aspect of the present disclosure, or a pharmaceutical composition according to the third aspect of the present disclosure, in the manufacture of a medicament for the prevention and/or treatment of pain and a disease associated with pain.

[0092] In another preferred embodiment, the MOR receptor agonist mediated disease is selected from the group consisting of pain, immune dysfunction, inflammation, esophageal reflux, neurological and psychiatric diseases, urological and reproductive diseases, cardiovascular diseases, and respiratory diseases, preferably pain.

[0093] In another preferred embodiment, the pain is selected from the group consisting of postoperative pain, pain caused by cancer, neuropathic pain, traumatic pain, and pain caused by inflammation.

[0094] In another preferred embodiment, the cancer is selected from the group consisting of breast cancer, endometrial cancer, cervical cancer, skin cancer, prostate cancer, ovarian cancer, fallopian tube tumor, ovarian tumor, hemophilia, and leukemia.

[0095] In a seventh aspect, the present disclosure provides a method for preventing and/or treating a disease mediated by an MOR receptor agonist, which comprises: administering to a patient in need thereof a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate, or prodrug according to the first or second aspect of the present disclosure, or a pharmaceutical composition according to the third aspect of the present disclosure. disclosure.

[0096] In an eighth aspect, the present disclosure provides a method for the prevention and/or treatment of pain and a disease associated with pain, which includes: administering to a patient in need thereof a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate or prodrug in accordance with the first or second aspect of the present disclosure, or a pharmaceutical composition in accordance with the third aspect of the present disclosure.

[0097] In another preferred embodiment, the MOR receptor agonist mediated disease is selected from the group consisting of pain, immune dysfunction, inflammation, esophageal reflux, neurological and psychiatric diseases, urological and reproductive diseases, cardiovascular diseases, and respiratory diseases, preferably pain.

[0098] In another preferred embodiment, the pain is selected from the group consisting of postoperative pain, pain caused by cancer, neuropathic pain, traumatic pain, and pain caused by inflammation.

[0099] In another preferred embodiment, the cancer is selected from the group consisting of breast cancer, endometrial cancer, cervical cancer, skin cancer, prostate cancer, ovarian cancer, fallopian tube tumor, ovarian tumor, hemophilia, and leukemia.

[0100] In a ninth aspect, the present disclosure provides a method for preparing a compound represented in formula (II), which includes the following steps:

[0101] carrying out reductive amination of the compound shown in formula (I-1) and the compound shown in formula (I-2) to obtain the compound shown in formula (I),

[0102] where the groups in each formula are as defined above.

[0103] In another preferred embodiment, the compound represented in formula (I-1) is .

[0104] In another preferred embodiment, the compound represented in formula (I-2) is .

[0105] In another preferred embodiment, the reductive amination is carried out in a reaction system containing an inert solvent, a Lewis acid and a reducing agent.

[0106] In another preferred embodiment, the inert solvent is C _1-4 alkyl alcohol, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, or a combination thereof.

[0107] In another preferred embodiment, the Lewis acid is titanium isopropoxide.

[0109] In another preferred embodiment, the reducing agent is selected from the group consisting of tetrabutylammonium borohydride, sodium malonyloxyborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, and borane.

[0109] In a tenth aspect, the present disclosure provides a method for preparing a compound represented in formula (II), which includes the following steps:

[0110] carrying out reductive amination of the compound shown in formula (I-3) and the compound shown in formula (I-4) to obtain the compound shown in formula (I),

[0111] where the groups in each formula are as defined above.

[0112] In another preferred embodiment, the compound represented in formula (I-3) is .

[0113] In another preferred embodiment, the compound represented in formula (I-4) is .

[0114] In another preferred embodiment, the reductive amination is carried out in a reaction system containing an inert solvent and a reducing agent.

[0115] In another preferred embodiment, the inert solvent is C _1-4 alkyl alcohol, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, or a combination thereof.

[0116] In another preferred embodiment, the reducing agent is selected from the group consisting of tetrabutylammonium borohydride, sodium malonyloxyborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, and borane.

[0117] In an eleventh aspect, the present disclosure provides an intermediate of a ring-substituted oxaspirocyclic pyrrolopyrazole derivative, wherein the intermediate has the structure represented by formula (I-2):

(I-2),

[0118] in which Y ₁ , Y ₂ , Y ₃ , Y ₄ , W, R _a , R _b , R _c , R _d and m are as defined in the first aspect of the present disclosure.

[0119] In some embodiments, the compound represented by formula (I-2) has the structure represented by formula (I-2a):

(I-2a).

[0120] In some embodiments, the compound represented by formula (I-2) has the structure represented by formula (I-2b):

(I-2b).

[0121] In some embodiments, R _a and R _b are each independently hydrogen or C _1-3 alkyl (preferably methyl); or R _a and R _b together with their associated carbon atom form a three to six membered saturated monocyclic ring (preferably a cyclopropyl ring, cyclobutyl ring or cyclopentyl ring).

[0122] In some embodiments, R _c is hydrogen, halogen (preferably F), C _1-3 alkyl (preferably methyl, ethyl, or n-propyl), C _1-3 alkoxy (preferably methoxy), or halogenated C _1-3 alkyl (preferably trifluoromethyl).

[0123] In some embodiments, the compound represented in formula (I-2) is selected from the following structures:

[0124] In a twelfth aspect, the present disclosure provides a process for preparing a compound represented in formula I-2b according to the eleventh aspect of the present disclosure, which includes the following step:

[0125] S1: carrying out cyclization of the compound represented in formula I-2b ₅ to obtain the compound represented in formula I-2b.

[0126] In some embodiments, in step S1, the cyclization is carried out in a solution of n-butyllithium in THF.

[0127] In some embodiments, Compound I-2b ₅ is a known substance (e.g., CAS No. 1310249-96-7 or CAS No. 2301048-78-0) or is prepared with reference to relevant published documents or by conventional reactions known to those skilled in the art.

[0128] In some other embodiments, obtaining a compound represented in formula I-2b ₅ includes the following steps:

[0129] S2: carrying out the hydrolysis reaction of the compound represented in formula I-2b ₄ to obtain the compound represented in formula I-2b ₅ .

[0130] In some embodiments, in step S2, the hydrolysis reaction is carried out in a solvent (e.g., methanol/water, THF/methanol, THF/methanol/water) under inorganic alkali catalysis (e.g., sodium hydroxide, lithium hydroxide, potassium hydroxide).

[0131] In some embodiments, the implementation of obtaining a compound represented in formula I-2b ₄ includes the following steps:

[0132] S3: carrying out an addition reaction of a compound represented by formula I-2b ₃ and pyrazole to obtain a compound represented by formula I-2b ₄ .

[0133] In some embodiments, in step S3, the addition reaction is carried out in an alkaline system (eg, DBU, potassium carbonate, etc.) in an organic solvent (eg, acetonitrile, DMF, etc.).

[0134] In some embodiments, the compound represented in formula I-2b ₃ is a known substance (e.g., ethyl (E)-but-2-enoate, ethyl (E)-hex-2-enoate, etc.) or can be obtained with reference to relevant published documents or using conventional reactions known to those skilled in the art.

[0135] In some other embodiments, the implementation of the compound represented in formula I-2b ₃ is obtained using the following step:

[0136] where R _e represents C _1-6 alkyl, and preferably methyl, ethyl, propyl or isopropyl,

[0137] S4: Reacting a compound represented by formula I-2b ₁ with a compound represented by formula I-2b ₂ to obtain a compound represented by formula I-2b ₃ .

[0138] In some embodiments, in step S4, the reaction is carried out in the presence of DBU/lithium chloride in an organic solvent (eg, acetonitrile, THF).

[0139] In some embodiments, the compound represented in formula I-2b ₁ is a known substance (for example, cyclopropanecarboxaldehyde, cyclopentanecarboxaldehyde, propionaldehyde, isobutyraldehyde, pivalaldehyde, 3-methylbutyraldehyde, cyclobutanone, etc.) or can be obtained with reference to relevant published documents or using conventional reactions known to those skilled in the art.

[0140] In some embodiments, the compound represented in formula I-2b ₂ is a known substance (for example, triethylphosphonoacetate) or can be obtained with reference to relevant published documents or using conventional reactions known to those skilled in the art.

[0141] It should be understood that within the scope of the present disclosure, the above technical features of the present disclosure and the technical features specifically described below (eg, embodiments) may be combined with each other to create new or preferred technical solutions. Due to space limitations, they will not be repeated in this document.

DESCRIPTION OF EMBODIMENTS

[0142] Through extensive and in-depth research, the inventors have surprisingly found that such azabicyclic ring-substituted oxaspirocyclic derivatives not only have excellent analgesic activity, but also good measurement error. In addition, the compounds of the present disclosure have excellent pharmacokinetic properties. Thus, these compounds are expected to be developed as drugs for the treatment and prevention of pain and diseases associated with pain. On this basis, the inventors have made the present disclosure.

[0143] Definition of terms

[0144] As used herein, the term "alkyl" refers to a linear and branched saturated aliphatic hydrocarbyl group. C _1-10 alkyl refers to an alkyl group having 1 to 10 carbon atoms, C _1-8 alkyl is preferred, C _1-6 alkyl is more preferred, C _1-4 alkyl is more preferred, and C _1-3 alkyl is even more preferred, which have similar definitions. Examples of alkyl more preferably include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2 ,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl yl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, their various branched isomers, etc.

[0145] As used herein, the term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group having carbon-carbon double bonds (C=C), which preferably contains 2 to 10 carbon atoms (C _2-10 ), more preferably contains 2 to 6 carbon atoms (C _2-6 ), and even more preferably contains 2 to 4 carbon atoms (C _2-4 ). Examples of alkenyl include, but are not limited to, vinyl, propenyl, isopropenyl, n-butenyl, isobutenyl, pentenyl, hexenyl, and the like.

[0146] As used herein, "alkynyl" refers to a linear and branched unsaturated aliphatic hydrocarbon group having carbon-carbon triple bonds that preferably contains 2 to 10 carbon atoms (C _2-10 ), more preferably contains 2 to 6 carbon atoms (C _2-6 ), and even more preferably contains 2 to 4 carbon atoms (C _2-4 ). Examples of alkynyl include, but are not limited to, ethynyl, propynyl, n-butynyl, isobutynyl, pentynyl, hexynyl, etc.

[0147] As used herein, the terms "cycloalkyl" and "cycloalkyl ring" may be used interchangeably, and both refer to saturated or partially unsaturated monocyclic or polycyclic hydrocarbon groups. "C _3-8 cycloalkyl" refers to a cyclic hydrocarbyl group having 3 to 8 carbon atoms, and preferred is C _3-6 cycloalkyl, which has a similar definition. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc., and cyclopropyl, cyclopentyl, cyclohexenyl are preferred.

[0148] As used herein, the term "spirocyclic ring" refers to a polycyclic group in which the monocyclic rings share one carbon atom (called a spiro atom), these monocyclic rings may contain one or more double bonds, but none of the rings has a fully conjugated π electron system. According to the number of rings, spiro rings are divided into spiro bicyclic rings and spiro polycyclic rings, with spiro bicyclic rings being preferred. A four-membered/five-membered, five-membered/five-membered or five-membered/six-membered spirobicyclic ring is more preferred. Examples include:

.

[0149] As used herein, the term "spiro-heterocyclic ring" refers to a polycyclic hydrocarbon in which the monocyclic rings share one atom (referred to as a spiro atom), one or two ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O) _n (where n is an integer from 0 to 2), and the other atoms in the ring are carbon atoms. These rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. According to the number of rings, spiro-heterocyclic rings are divided into bicyclic spiro-heterocyclic rings and polycyclic spiro-heterocyclic rings, with bicyclic spiro-heterocyclic rings being preferred. A four-membered/five-membered, five-membered/five-membered or five-membered/six-membered bicyclic spiro-heterocyclic ring is more preferred. Examples include:

.

[150] As used herein, the term "bridged ring" refers to a polycyclic group in which two or more common carbon atoms are shared, where the common carbon atoms are called bridging carbon atoms, and the two bridging carbon atoms can be connected by a carbon chain or bond, which is called a bridge. This bridging ring may contain one or more double bonds, but none of the rings has a fully conjugated π electron system. Bicyclic or tricyclic bridged rings are preferred. Examples include:

.

[0151] As used herein, the term "bridged heterocyclic ring" refers to a polycyclic group in which two or more common atoms are shared, one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O) _n (where n is an integer from 0 to 2), and the other ring atoms are carbon atoms. A bridged heterocyclic ring may contain one or more double bonds, but none of the rings has a fully conjugated π electron system. Bridged heterobicyclic or heterotricyclic rings are preferred. Examples include:

.

[0152] As used herein, the term "eight to ten membered bicyclic ring" refers to a bridged ring containing two rings with 8 to 10 ring atoms, and the bicyclic ring can be a saturated all-carbon bicyclic ring or a partially unsaturated all-carbon bicyclic ring. Examples of an eight to ten membered bicyclic ring include, but are not limited to,

.

[0153] As used herein, the term "eight to ten membered heterocyclic ring" refers to a bridged heterocyclic ring containing two rings with 8 to 10 ring atoms, in which one, two, three, four, or five carbon atoms in the ring are replaced by heteroatoms selected from nitrogen, oxygen, or sulfur. Examples of the eight to ten membered heterobicyclic ring include, but are not limited to, tetrahydroquinoline ring, tetrahydroisoquinoline ring, decahydroquinoline ring, etc.

[0154] As used herein, the term "C _1-8 alkoxy" refers to -O-(C _1-8 alkyl), in which alkyl is as defined above. Preferred is C _1-6 alkoxy, preferred is C _1-4 alkoxy, and more preferred is C _1-3 alkoxy. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, isobutoxy, pentyloxy, etc.

[0155] As used herein, the term "C _3-8 cycloalkoxy" refers to -O-(C _3-8 cycloalkyl), in which cycloalkyl is as defined above. Preferred is C _3-6 cycloalkoxy. Examples of cycloalkoxy include, but are not limited to, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, etc.

[0156] As used herein, the terms "C _6-10 aryl" and "C _6-10 aromatic ring" can be used interchangeably, both referring to an all-carbon monocyclic or fused polycyclic (i.e., the rings share adjacent pairs of carbon atoms) group having a conjugated π electron system and refer to an aryl group having 6 to 10 carbon atoms. Phenyl and naphthyl are preferred, and phenyl is more preferred.

[0157] As used herein, the term "one bond" refers to a covalent bond through which two groups are linked.

[0158] As used herein, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0159] As used herein, the term "halogenated" refers to one or more (eg, one, two, three, four, or five) hydrogen atoms in the group being replaced by a halogen.

[0160] For example, "halogenated C _1-10 alkyl" means that the alkyl group is substituted with one or more (eg, one, two, three, four or five) halogen atoms, in which alkyl is as defined above. Halogenated C _1-8 alkyl is preferred, more preferred is halogenated C _1-6 alkyl, more preferred is halogenated C _1-4 alkyl, and even more preferred is halogenated C _1-3 alkyl. Examples of halogenated alkyl include, but are not limited to, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, etc.

[0161] In another example, "halogenated C _1-10 alkoxy" means that the alkoxy group is substituted with one or more (eg, one, two, three, four or five) halogen atoms, in which the alkoxy is as defined above. Halogenated C _1-8 alkoxy is preferred, more preferred is halogenated C _1-6 alkoxy, more preferred is halogenated C _1-4 alkoxy, and even more preferred is halogenated C _1-3 alkoxy. Halogenated alkoxy groups include, but are not limited to, trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethoxy, etc.

[0162] In another example, "halogenated C _3-8 cycloalkyl" means that the cycloalkyl group is substituted with one or more (eg, one, two, three, four, or five) halogen atoms, wherein the cycloalkyl is as defined above. Preferred is C _3-6 cycloalkyl. Examples of halogenated cycloalkyl include, but are not limited to, trifluorocyclopropyl, monofluorocyclopropyl, monofluorocyclohexyl, difluorocyclopropyl, difluorocyclohexyl, etc.

[0163] As used herein, the term "deuterated C _1-8 alkyl" means that the alkyl group is substituted with one or more (eg, one, two, three, four, or five) deuterium atoms, in which alkyl is as defined above. Preferred is deuterated C _1-6 alkyl, and more preferred is deuterated C _1-3 alkyl. Examples of the deuterated alkyl group include, but are not limited to, mono-deuteromethyl, mono-deuteroethyl, di-deuteromethyl, di-deuteroethyl, tri-deuteromethyl, tri-deuteroethyl, etc.

[0164] As used herein, "amino" refers to NH ₂ , "cyano" refers to CN, "nitro" refers to NO ₂ , "benzyl" refers to -CH ₂ -phenyl, "oxo" refers to =O, "carboxyl" refers to -C(O)OH, "acetyl" refers to -C(O)CH ₃ , "hydroxymethyl" refers to -CH ₂ OH, "hydroxyethyl" refers to -CH ₂ CH ₂ OH or -CHOHCH ₃ , "hydroxyl" refers to -OH, "thiol" refers to SH, and "cyclopropylidene" has the following structure: .

[0165] As used herein, the terms "heteroaryl ring" and "heteroaryl" can be used interchangeably, both refer to a group having 5 to 10 ring atoms, 6, 10, or 14 shared π electrons in an array of rings, and 1 to 5 heteroatoms in addition to carbon atoms. "Heteroatom" refers to nitrogen, oxygen or sulfur. Five or six membered monocyclic heteroaryl or eight to ten membered bicyclic heteroaryl is preferred.

[0166] As used herein, the term "three-seven-membered (four-seven-membered) saturated monocyclic ring" refers to a saturated all-carbon monocyclic ring containing 3 to 7 ring atoms, and a three- to six-membered saturated monocyclic ring is preferred. Examples of saturated monocyclic rings include, but are not limited to, cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclohexyl ring, cycloheptyl ring, cyclooctyl ring, etc.

[0167] As used herein, the term "three-seven-membered (four-seven-membered) saturated monoheterocyclic ring" means that one, two or three carbon atoms in a three-seven-membered saturated monocyclic ring are replaced by selected heteroatoms selected from nitrogen, oxygen, or S(O) _t (where t is an integer from 0 to 2), the -OO-, -OS- or -SS- ring fragment is not included, and the other atoms in the ring are carbon atoms. Preferred is a four to six membered saturated monoheterocyclic ring, and more preferred is a five or six membered saturated monoheterocyclic ring. An example of a saturated monoheterocyclic ring includes, but is not limited to, propylene oxide, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, pyrroline, oxazolidine, piperazine, dioxolane, dioxane, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, etc.

[0168] As used herein, the terms "five or six membered monocyclic heteroaryl ring" and "five or six membered monocyclic heteroaryl ring" may be used interchangeably and both refer to a monocyclic heteroaryl ring having 5 or 6 ring atoms, which includes, but is not limited to, thiophene ring, furan ring, thiazole ring, imidase ring, oxazole ring, pyrrole new ring, pyrazole ring, triazole ring, 1,2,3-triazole ring, 1,2,4-triazole ring, 1,2,5-triazole ring, 1,3,4-triazole ring, tetrazole ring, isoxazole ring, oxadiazole ring, 1,2,3-oxadiazole ring, 1,2,4-oxadiazole ring, 1,2,5-oxadia ash ring, 1,3,4-oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring, etc.

[0169] As used herein, the terms "eight to ten membered bicyclic heteroaryl ring" and "eight to ten membered bicyclic heteroaryl ring" can be used interchangeably, and both refer to a bicyclic heteroaryl ring containing 8 to 10 ring atoms, including, but not limited to, benzofuran, benzothiophene, indole, isoindole, quinoline, isoquinoline, indazole, benzothiazole, benzimidazole, quinazoline, quinoxaline, cinnoline, phthalazine, pyrido[3,2-d]pyrimidine, pyrido[2,3-d]pyrimidine, pyrido[3,4-d]pyrimidine, pyrido[4,3-d]pyrimidine, 1,8-naphthyridine, 1,7-naphthyridine, 1,6-naph thyridine, 1,5-naphthyridine.

[0170] As used herein, the term "substituted" refers to one or more hydrogen atoms, preferably from one to five hydrogen atoms, and more preferably from one to three hydrogen atoms in a group, each independently substituted with an appropriate number of substituent groups. That is, substituent groups are found only in their possible chemical positions, and those skilled in the art can determine (experimentally or theoretically) possible or impossible substitutions without any effort. For example, the bonding of an amino or hydroxyl group having free hydrogen atoms to carbon atoms having unsaturated (eg, olefinic) bonds may be unstable.

[0171] Unless otherwise indicated, the phrase "each substituent group is independently selected from..." as used herein means that when more than one hydrogen atom in a group is substituted with substituent groups, the substituent groups may be of the same type or different types, and the selected substituent substituent groups are independent in terms of their types.

[0172] Unless otherwise indicated, the phrase "... are the same or different, and each independently ..." as used herein means that when there is more than one identical substituent group in a general formula, the groups may be the same or different and independent in their types. For example, provided that L is (CR ₀₁ R ₀₂ ) _s when s is 2, that is, L is (CR ₀₁ R ₀₂ )-(CR ₀₁ R ₀₂ ), where the two groups R ₀₁ or R ₀₂ may be the same or different and independent in their types. For example, L may be C(CH ₃ )(CN)-C(CH ₂ CH ₃ )(OH), C(CH ₃ )(CN)-C(CH ₃ )(OH), or C(CN)(CH ₂ CH ₃ )-C(OH)(CH ₂ CH ₃ ).

[0173] Unless otherwise indicated, any of the groups described herein may be substituted or unsubstituted. Если указанная выше группа является замещенной, при этом замещающие группы предпочтительно представляют собой от одной до пяти групп, каждая из которых независимо выбрана из CN, галогена, C _1-10 алкила (предпочтительно C _1-6 алкила и более предпочтительно C _1-3 алкила), C _1-10 алкоксила (предпочтительно C _1-6 алкоксила и более предпочтительно C _1-3 алкоксила), галогенированного C _1-8 алкила (предпочтительно галогенированного C _1-6 алкила и более предпочтительно галогенированного C _1-3 алкила), C _3-8 циклоалкила (предпочтительно C _3-6 циклоалкила), галогенированного C _1-8 алкоксила (предпочтительно галогенированного C _1-6 алкоксила и более предпочтительно галогенированного C _1-3 алкоксила), C _1-8 алкилзамещенного амидо, амидо, галогенированного C _1-8 C _1-8 алкилзамещенного амидо, ацетила, гидроксила, гидроксиметила, гидроксиэтила, карбоксила, нитро, C _6-10 арила (предпочтительно фенила), C _3-8 циклоалкоксила (предпочтительно C _3-6 циклоалкоксила), C _2-10 алкенила (предпочтительно C _2-6 алкенила и более предпочтительно C _2-4 алкенила), C _2-10 алкинила (предпочтительно C _2-6 алкинила и более предпочтительно C _2-4 алкинила), -CONR _a0 R _b0 , -C(O)O(C _1-10 алкила) (предпочтительно -C(O)O(C _1-6 алкила) и более предпочтительно -C(O)O(C _1-3 алкила)), -CHO, -OC(O)(C _1-10 алкила) (предпочтительно -OC(O)(C _1-6 алкила) и более предпочтительно -OC(O)(C _1-3 алкила)), -SO2(C _1-10 алкила) (предпочтительно -SO ₂ (C1-6 алкила) и более предпочтительно -SO2(C _1-3 алкила)), -SO2(C _6-10 арила) (предпочтительно -SO ₂ (C ₆ арила), а именно -SO ₂ -фенила), -CO(C _6-10 арила) (предпочтительно -CO(C ₆ арила), а именно -CO-фенила), четырех-шестичленного насыщенного или ненасыщенного моногетероциклического кольца, четырех-шестичленного насыщенного или ненасыщенного моноциклического кольца, пяти- или шестичленного моноциклического гетероарильного кольца, восьми-десятичленного бициклического гетероарильного кольца, спирокольца, спирогетероциклического кольца, мостикового кольца и мостикового гетероциклического кольца, в которой каждый из R _a0 и R _b0 независимо представляет собой водород или C _1-3 алкил.

[0174] Each substituent group described above may itself also be substituted by the groups described herein.

[0175] When the four-to-six-membered (five- or six-membered) saturated monoheterocyclic ring described herein is substituted, the substituent groups may be in their possible chemical positions. Representative substitutions for typical monoheterocyclic rings are shown below:

where "substituent" refers to the various substituent groups described herein, and " " refers to bonding with other atoms.

[0176] Unless otherwise indicated, when the four to six membered saturated monoheterocyclic ring described herein is a substituent group, it may also be substituted or substituted with one, two or three substituent groups selected from the group consisting of halo, hydroxyl, C_1-3 alkyl, O=, NR_a0R_b0, hydroxymethyl, hydroxyethyl, carboxyl, -C(O)O(C_1-3 alkyl), acetyl, halogenated C_1-3 alkyl, C_1-3 alkoxy, C_3-6 cycloalkyl, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, oxazolidine, piperazine, dioxolane, dioxane, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, thiophene ring, N-alkylpyrrole ring, furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, tetrazole ring, isoxazole ring, oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring and pyrazine ring, in which each of R_a0 and R_b0 is independently hydrogen or C_1-3 alkyl.

[0177] A "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0178] "Pharmaceutical acid addition salts" refers to salts formed with inorganic or organic acids that can retain the biological effectiveness of free alkali without causing other side effects.

[0179] "Pharmaceutical base addition salts" include, but are not limited to, salts such as sodium salts, potassium salts, calcium salts, and magnesium salts, inorganic alkalis, and salts such as ammonium salts, triethylamine salts, lysine salts, and organic alkali arginine salts.

[0180] "Solvate" as described herein refers to a complex formed by a compound of the present disclosure and a solvent. Compounds either react in solvents or precipitate or crystallize from solvents. For example, a complex formed by a compound and water is called a "hydrate". Solvates of the compound represented in formula (I) are within the scope of this disclosure.

[0181] The compound represented in formula (I) or formula (II) of the present disclosure may contain two or more chiral centers that are present in various optically active forms. The stereoisomer of the compound represented in formula (I) or formula (II) of the present disclosure may be an enantiomer or a diastereomer. The compound represented in formula (I) or in formula (II) may be present in the form of a single optically pure specific stereoisomer. For example, the compound may be present in the form of an enantiomer or diastereomer, or in the form of a mixture of two stereoisomers. For example, the compound may be present in the form of a mixture of enantiomers, such as a racemic mixture, or in the form of a mixture of diastereomers, or in the form of a mixture of an enantiomer and a diastereomer. The enantiomer can be obtained using separation methods existing in the art, such as crystallization and chiral chromatography. The diastereomer can be obtained using existing separation methods such as crystallization and preparative chromatography. Enantiomers or diastereomers of the compound represented in formula (I) or formula (II) of the present disclosure and mixtures of these stereoisomers are within the scope of the present disclosure.

[0182] The present disclosure further relates to prodrugs based on the above compounds. Prodrugs include known amino protecting groups and carboxy protecting groups which are hydrolyzed under physiological conditions or released via enzymatic reactions to give the parent compounds. For specific prodrug preparation methods, see Saulnier, M.G.; Frennesson, D. B.; Deshpande, M.S.; Hansel, S.B. and Vysa, D.M. Bioorg. Med. Chem Lett. 1994, 4, 1985-1990; and Greenwald. R.B.; Choe, Y. H.; Conover, C.D.; Shum, K.; Wu, D.; Royzen, M.J. Med. Chem. 2000, 43, 475.

[0183] Generally, a compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate, mixture of diastereomers, solvate, or prodrug of the present disclosure and one or more pharmaceutical carriers thereof, and one or more pharmaceutical carriers, may be formulated into suitable formulations for administration. These formulations are suitable for oral administration, rectal administration, topical administration, intraoral administration, and other parenteral (eg, subcutaneous, intramuscular, intravenous, etc.) administration. For example, formulations suitable for oral administration include capsules, tablets, granules, syrups, etc. The compound of the present disclosure contained in these formulations may be a solid powder or particles, a solution or suspension in an aqueous or non-aqueous liquid, or a water-in-oil or oil-in-water emulsion, etc. The above formulations may be prepared from the active compound and one or more carriers or excipients by general pharmaceutical methods. The carriers mentioned above must be compatible with the active compound or other excipients. For solid formulations, non-toxic carriers commonly used include, but are not limited to, mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose, and the like. For liquid formulations, carriers commonly used include water, saline, aqueous glucose, ethylene glycol, polyethylene glycol, etc. The active compound may form a solution or suspension with the above carrier.

[0184] Compositions of the present disclosure are prepared, dosed, and administered in accordance with medical practice. The "therapeutically effective amount" of the compound to be administered is determined by such factors as the particular disease being treated, the individual being treated, the cause of the disorder, the purpose of the drug, the route of administration, etc.

[0185] As used herein, the term "therapeutically effective amount" refers to an amount of a compound of the present disclosure that will elicit a biological or medical response in an individual, such as reducing or inhibiting enzyme or protein activity or ameliorating symptoms, alleviating a disorder, slowing or delaying disease progression, and preventing disease.

[0186] A therapeutically effective amount of a compound or a pharmaceutically acceptable salt, solvate or stereoisomer thereof of the present disclosure contained in a pharmaceutical composition of the present disclosure is preferably 0.1 mg to 5 g/kg (body weight).

[0187] As used herein, the term "pharmaceutically acceptable carrier" refers to non-toxic, inert, and solid or semi-solid substances or liquid excipients, diluents, encapsulating materials, or adjuvants, or excipients of any type, which are compatible with patients, preferably mammals, and more preferably humans, and are suitable for delivering the active reagent to the target without termination of the reagent.

[0188] As used herein, the term "patient" refers to an animal, preferably a mammal, and more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, which include cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice or rats, pigs, and humans.

[0189] As used herein, the term "treatment" refers to alleviating, slowing the progression, attenuating, preventing, or maintaining an existing disease or disorder (eg, cancer). Treatment also includes recovery from one or more symptoms of the disease or disorder, prevention of development, or relief to a certain extent.

[0190] Methods for obtaining

[0191] The experimental method described in the following examples without imposing specific conditions generally follows conventional conditions, such as those described in Sambrook, et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or conditions recommended by the manufacturers.

[0192] Unless otherwise indicated, terms used herein have the same meanings as terms known to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applicable to the present disclosure.

[0193] Methods for preparing compounds of the present disclosure:

[0194] The compounds represented in formula (I) and formula (II) can be obtained using existing methods such as the following methods, methods equivalent to the following methods, and the methods described in the examples. In the following preparations, the raw starting compound may be in the form of a salt, and the salt may be any pharmaceutically acceptable salt as shown for the compounds represented in formula (I) and formula (II) of the present disclosure.

[0195] Reaction scheme (a1)

[0196] In each formula of the above scheme, all groups are as defined above.

[0197] In particular, the compound represented in formula (I) can be obtained by the following method: reductive amination is carried out on the compound represented in formula (I-1) and the compound represented in formula (I-2) to obtain the compound represented in formula (I). The reductive amination is carried out in an inert solvent, wherein the compound represented by formula (I-1), the compound represented by formula (I-2), and the Lewis acid are reacted at a certain temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, and more preferably 20°C to 60°C) for a period of time (e.g., 0.5 hour to 48 hour. , preferably 5 hours to 24 hours), then a reducing agent is added to the reaction solution, and the reaction solution is reacted at a certain temperature (for example, from -20°C to 80°C, preferably from 0°C to 60°C, and more preferably from 20°C to 60°C) for a period of time (for example, from 0.5 hours to 48 hours, preferably from 0.5 hours to 5 hours), to obtain the compound shown in formula (I). The inert solvent and reducing agent may be known in the art, and the reducing agent may be selected from tetrabutylammonium borohydride, sodium malonyloxyborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, borane, etc. The Lewis acid may be titanium isopropoxide, etc. The inert solvent may be selected from C _{1- 4} alkyl alcohol, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, or combinations thereof.

[0198] Reaction scheme (b1)

[0199] In each formula of the above scheme, all groups are as defined above.

[0200] In particular, the compound shown in formula (I) can also be obtained by the following method: reductive amination is carried out on the compound shown in formula (I-3) and the compound shown in formula (I-4) to obtain the compound shown in formula (I). The reductive amination is carried out in an inert solvent, and the compound represented by formula (I-3), the compound represented by formula (I-4), and the reducing agent are reacted at a certain temperature (for example, from -20°C to 80°C, preferably from 0°C to 60°C, and more preferably from 20°C to 60°C) for a period of time (for example, from 0.5 hours to 48 hours, preferably from 0.5 hours to 5 hours) to obtain the compound represented in formula (I). The inert solvent and reducing agent may be known in the art, and the reducing agent may be selected from tetrabutylammonium borohydride, sodium malonyloxyborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, borane, etc. The inert solvent may be selected from C _1-4 alkyl alcohol, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, or combinations thereof.

[0201] Reaction scheme (a2)

[0202] In each formula of the above solution, all groups are as defined above.

[0203] Specifically, the compound shown in formula (II) can be obtained by the following method: reductive amination is carried out on the compound shown in formula II-1 and the compound shown in formula II-2 to obtain the compound shown in formula (II). Reductive amination is carried out in an inert solvent, and the compound represented in formula (II-1), the compound represented in formula (II-2), and the Lewis acid are reacted at a certain temperature (for example, from -20°C to 80°C, preferably from 0°C to 60°C, and more preferably from 20°C to 60°C) for a period of time (for example, from 0.5 hours to 48 hours. , preferably 5 hours to 24 hours), then a reducing agent is added to the reaction solution, and the reaction solution is reacted at a certain temperature (for example, from -20°C to 80°C, preferably from 0°C to 60°C, and more preferably from 20°C to 60°C) for a period of time (for example, from 0.5 hours to 48 hours, preferably from 0.5 hours to 5 hours), to obtain the compound shown in formula (II). The inert solvent and reducing agent may be known in the art, and the reducing agent may be selected from tetrabutylammonium borohydride, sodium malonyloxyborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, borane, etc. The Lewis acid may be titanium isopropoxide, etc. The inert solvent may be selected from C _{1- 4} alkyl alcohol, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, or combinations thereof.

[0204] Reaction scheme (b2)

[0205] In each formula of the above scheme, all groups are as defined above.

[0206] In particular, the compound shown in formula (II) can also be obtained by the following method: reductive amination is carried out on the compound shown in formula II-3 and the compound shown in formula II-4 to obtain the compound shown in formula (II). The reductive amination is carried out in an inert solvent, and the compound represented by formula (II-3), the compound represented by formula (II-4), and the reducing agent are reacted at a certain temperature (for example, from -20°C to 80°C, preferably from 0°C to 60°C, and more preferably from 20°C to 60°C) for a period of time (for example, from 0.5 hours to 48 hours, preferably from 0.5 hours to 5 hours) to obtain the compound represented in formula (II). The inert solvent and reducing agent may be known in the art, and the reducing agent may be selected from tetrabutylammonium borohydride, sodium malonyloxyborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, borane, etc. The inert solvent may be selected from C _1-4 alkyl alcohol, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, or combinations thereof.

[0207] Compounds containing an amino group, a carboxyl group, or a hydroxyl group, which are used in the present disclosure, can be obtained from compounds protected by protecting groups commonly used for groups if necessary, and after the reaction process of the above reaction scheme, a known deprotection can be performed.

[0208] Compared with the prior art, the present disclosure mainly has the following advantages.

[0209] The present disclosure provides a range of azabicyclic ring substituted oxaspirocyclic derivatives with novel structures that have high cAMP inhibitory activity (EC ₅₀ is 1 nM to 100 nM, more preferably 1 nM to 50 nM, and most preferably 1 nM to 10 nM) while having high Emax values (above 50% , preferably above 80%) and have excellent analgesic effects. In addition, the compounds of the present disclosure have small Emax values with respect to β-arrestin (less than 50%, more preferably less than 30%, and most preferably less than 10%) and have a good measurement error. Thus, the compounds of the present disclosure can be formulated into drugs for the treatment and prevention of pain and diseases associated with pain.

[0210] The present disclosure will be further described below with reference to specific examples. It should be understood that these examples are only for the purpose of describing the present disclosure and not limiting the scope of the present disclosure. The experimental method described in the following examples without the introduction of specific conditions, usually corresponds to the usual conditions or conditions recommended by the manufacturers. Unless otherwise stated, percentages and parts are calculated based on weight. Unless otherwise indicated, terms used herein have the same meanings as terms known to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applicable to the present disclosure.

[0211] Reagents and instruments

[0212] ¹ HNMR: Bruker AVANCE-400 NMR spectrometer, internal standard: tetramethylsilane (TMS);

[0213] LC-MS: Agilent 1290 HPLC System/6130/6150 MS LC/MS spectrometer (manufacturer: Agilent), column: Waters BEH/CHS, 50×2.1 mm, 1.7 µm.

[0214] Preparative High Performance Liquid Chromatography (pre-HPLC): GX-281 (manufacturer: Gilson).

[0215] Supercritical liquid chromatography (SFC): 1260 Infinity (manufacturer: Agilent).

[0216] As used herein, DCE refers to 1,2-dichloroethane, THF refers to tetrahydrofuran, EA refers to ethyl acetate, PE refers to petroleum ether, DCM refers to dichloromethane, n-BuLi refers to n-butylium, HATU refers to 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate y, DMF refers to dimethylformamide, DMSO refers to dimethyl sulfoxide, DIEA or DIPEA refers to N,N-diisopropylethylamine, DBU refers to 1,8-diazabicyclo[5.4.0]undec-7-ene, and DIBAL-H refers to diisobutylaluminum hydride, and LiHMD refers to lithium bis(trimethylsilyl)amide.

[0217] As used herein, room temperature refers to approximately 20°C to 25°C.

[0218] Intermediate 1a

[0219] ( R )-2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)acetonitrile (10 g, 39 mmol, purchased from Shanghai Yucheng Pharmaceutical Technology Co., Ltd., CAS No. 1401031-38-6) and Raney nickel (1 g) were added to ethanol (200 ml) and ammonia water (30 ml) and the reaction solution was stirred at room temperature for 12 hours under the protection of hydrogen gas, followed by filtration and concentration under reduced pressure to give ( R )-2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)atamine (Intermediate 1a , 10 g, colorless oily liquid) in 99% yield. MS m/h (ESI): 261.2 [M+1].

[0220] Example 1.

N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-1)

[0221] In step 1: ethyl 1H-pyrazole-5-carboxylate (10 g, 7.14 mmol) was dissolved in DMF (100 ml), ethyl acrylate (10.7 g, 10.7 mmol) was added and the reaction solution was stirred at 100° C. overnight. EA (400 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using eluents (PE: EA=1: 1) to obtain compound 1-1 (1.2 g) with a yield of 7%. MS m/s (ESI): 241.1 [M+1].

[0222] In step 2: compound 1-1 (300 mg, 1.25 mmol) was dissolved in a solution of toluene (8 ml), potassium tert-butoxide (280 mg, 2.5 mmol) was added, and the reaction solution was stirred at 110° C. for 1 hour. hydrochloric acid, then washed with saturated sodium bicarbonate solution and dried over anhydrous sodium sulfate. The resulting organic phase was concentrated under reduced pressure to give compound 1-2 (130 mg). MS m/h (ESI): 195.1 [M+1].

[0223] In step 3: Compound 1-2 (130 mg) was dissolved in a methanol solution (8 ml), a 4 M solution of hydrogen chloride in methanol (1 ml) was added, and the reaction solution was stirred at 50° C. overnight. The resulting organic phase was concentrated under reduced pressure to give compound 1-3 (100 mg, crude product). MS m/h (ESI): 123.1 [M+1].

[0224] In Step 4: Compound 1-3 (100 mg, 0.82 mmol) was dissolved in a solution of DCE (8 ml), compound 1a (210 mg, 0.82 mmol) and titanium isopropoxide (1 ml) were added, and the reaction solution was heated to 60° C. to react overnight. After the reaction solution was cooled to room temperature, sodium borohydride (94 mg, 2.46 mmol) was added, and the reaction solution was stirred at room temperature for 1 hour. Water was added, the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% change in acetonitrile) to give compound H-1 (2.15 mg) in 0.7% yield. MS m/s (ESI): 367.1 [M+1]; ¹ H ЯМР (400 МГц, CDCl ₃ ): δ 8,56 (м, 1H), 8,12 (с, 1H), 7,68-7,64 (м, 1H), 7,46-7,45 (м, 1H), 7,32-7,29 (м, 1H), 7,17-7,13 (м, 1H), 5,92-5,89 (д, J=12 Гц, 1H), 4,26-4,20 (м, 2H), 4,06-4,01 (м, 1H), 3,72-3,69 (м, 2H), 2,87-2,74 (м, 2H), 2,64-2,43 (м, 6H), 2,35-2,29 (м, 3H), 2,10-2,03 (м, 1H), 1,91-1,84(м, 2H), 1,78-1,63(м, 3H), 1,24-1,21(м, 1H).

[0225] Example 2.

6-(2-methoxypropan-2-yl)-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-2)

[0226] In step 1: 4-chlorothiophenol (4.32 g, 30 mmol) was dissolved in methanol (100 ml), methyl chloroacetate (3.84 g, 31.5 mmol) and 5.4 M sodium methoxide in methanol solution (5.4 ml, 31.5 mmol) were added and the reaction solution was heated to 80° C. and stirred for 1 h. was taken, and the resulting residue was diluted with EA (150 ml), then washed with water (80 ml × 1), washed with brine (40 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE/EA=5/1) to give compound 2-1 (5.0 g, yellow oily liquid) in 77% yield. MS m/h (ESI): 217.3 [M+1].

[0227] In Step 2: Compound 2-1 (5 g, 23 mmol) was dissolved in acetic acid (60 ml), 30% hydrogen peroxide (2.45 g, 23 mmol) was slowly added with stirring, and the reaction solution was stirred at room temperature overnight. The reaction solution was concentrated, the resulting residue was diluted with EA (150 ml), then washed with water (70 ml×1), washed with saturated brine (40 ml×1), dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE/EA=3/1) to give compound 2-2 (4.2 g, yellow oily liquid) in 79% yield. MS m/h (ESI): 233.3 [M+1].

[0228] In step 3: Compound 2-2 (4.2 g, 18 mmol) and piperidine (1.85 g, 21.8 mmol) were dissolved in acetonitrile (60 ml) and isobutyraldehyde in acetonitrile solution (1.57 g, 21.8 mmol) was slowly added dropwise with stirring. The reaction solution was heated at 60°C and stirred for 5 hours. The reaction solution was concentrated, the resulting residue was diluted with EA (150 ml), then washed with water (70 ml × 1), washed with saturated brine (40 ml × 1), dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE/EA=3/1) to give compound 2-3 (1.68 g, yellow oily liquid) in 65% yield. MS m/h (ESI): 145.2 [M+1].

[0229] In step 4: compound 2-3 (1.68 g, 11.7 mmol) and iodomethane (2.49 g, 17.5 mmol) were dissolved in DMF (50 ml) and sodium hydride (0.7 g, 17.5 mmol) was added slowly dropwise with stirring. The reaction solution was stirred at room temperature for 3 hours. The reaction solution was diluted with EA (150 ml), then washed with water (70 ml×1), washed with brine (40 ml×1), dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE/EA=4/1) to give compound 2-4 (1.5 g, yellow oily liquid) in 81% yield. MS m/h (ESI): 159.2 [M+1].

[230] In step 5: a solution of acetonitrile (40 ml) and DBU (2.89 g, 19 mmol) was added to a sealed tube containing compound 2-4 (1.5 g, 9.5 mol) and pyrazole (1.29 g, 19 mmol), the reaction solution was stirred at 100 ° C for 18 hours, cooled to room temperature, concentrated, the resulting residue was EA (100 ml) was added, then washed with water (50 ml × 1), washed with brine (30 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE/EA=6/1) to give compound 2-5 (1.29 g, yellow oily liquid) in 60% yield. MS m/h (ESI): 227.3 [M+1].

[0231] In step 6: compound 2-5 (1.29 g, 5.7 mmol) was dissolved in a mixture of methanol (40 ml) and water (10 ml), sodium hydroxide (0.456 g, 11.4 mmol) was added and the reaction solution was stirred at room temperature for 1 hour until the reaction was completed. The reaction solution was concentrated, the resulting residue was dissolved in water (30 ml), 2 M hydrochloric acid was added to adjust the pH of the mixture to 3-4, the mixture was extracted with DCM (40 ml × 3), the combined organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure to obtain compound 2-6 (0.97 g, yellow oily liquid) with a yield of 80%. MS m/s (ESI): 213.2 [M+1].

[0232] In step 7: compound 2-6 (0.97 g, 4.56 mmol) was dissolved in dry THF (60 ml), the solution was cooled to -70°C under the protection of nitrogen gas, and a 2.5 M solution of butyllithium in THF (4.5 ml, 11.4 mmol) was slowly added dropwise. After completion of the dropwise addition, the reaction solution was slowly warmed to -40° C. and stirred for 45 minutes. Water (5 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (PE/EA=6/1) to give compound 2-7 (177 mg, yellow oily liquid) in 20% yield. MS m/h (ESI): 195.3 [M+1].

[0233] На шаге 8: соединение 2-7 (60 мг, 0,31 ммоль) и соединение 1а (80 мг, 0,31 ммоль) растворяли в ДХЭ (10 мл), добавляли изопропоксид титана (1 мл) и реакционный раствор перемешивали при 45°С в течение 18 ч. После охлаждения реакционного раствора до комнатной температуры добавляли боргидрид натрия (34 мг, 0,9 ммоль), реакционный раствор перемешивали в течение 3 ч., добавляли воду (5 мл), смесь перемешивали в течение 0,5 мин., фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной хроматографии (препаративная колонка: колонка С18 21,2 × 250 мм; система: 10 мМ NH ₄ HCO ₃ H ₂ O; длины волн: 254/214 нм; градиенты: 30%-60% изменений в ацетонитриле) с получением продукта H-2 (35 мг, бесцветное маслянистое вещество) с выходом 25,7%. MS m/h (ESI): 439.3 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,51-8,49 (м, 1H), 7,71-7,69 (м, 1H), 7,45-7,42 (м, 1H), 7,31-7,28 (м, 1H), 7,18-7,16 (м, 1H), 5,68 (дд, J=12,3, 1,8 Гц, 1H), 4,10-4,07 (м, 1H), 3,78 (тд, J=7,9, 4,8 Гц, 1H), 3,57 (дд, J=9,1, 2,8 Гц, 2H), 3,00 (с, 3H), 2,64 (м, 1H), 2,45-2,27 (м, 3H), 2,06-1,70 (м, 5H), 1,66-1,28 (м, 8H), 1,25 (д, J=7,6 Гц, 3H), 1,07 (д, J=7,1 Гц, 3H), 0,97-0,93 (м, 1H), 0,61-0,57 (м, 1H).

[0234] Example 3.

6-cyclopropyl-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-3)

[0235] In step 1: cyclopropanecarboxaldehyde (3.5 g, 50 mmol) was dissolved in acetonitrile (100 ml), triethylphosphonoacetate (11.8 g, 52.5 mmol) and lithium chloride (2.2 g, 52.5 mmol) were added, the reaction solution was cooled from 2 to 5 ° C, DBU (7.5 g, 52.5 mmol) was added slowly and the reaction The solution was kept at room temperature for 18 hours. The reaction solution was concentrated, and the resulting residue was diluted with EA (120 ml), then washed with water (80 ml × 1), washed with brine (40 ml × 1), dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE/EA=10/1) to give compound 3-1 (4.9 g, yellow oil) in 70% yield. MS m/h (ESI): 141.2 [M+1].

[0236] In step 2: a solution of acetonitrile (90 ml) and DBU (7.46 g, 52.5 mmol) was added to a sealed tube containing compound 3-1 (4.9 g, 35 mmol) and pyrazole (3.57 g, 52.5 mmol). The mixture was stirred at 120°C for 18 hours, cooled to room temperature and concentrated, the resulting residue was diluted with EA (100 ml), then washed with water (60 ml × 1), washed with brine (30 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE/EA=6/1) to give compound 3-2 (3.78 g, yellow oily liquid) in 52% yield. MS m/s (ESI): 209.3 [M+1].

[0237] In Step 3: Compound 3-2 (3.78 g, 18.2 mmol) was dissolved in a mixture of methanol (50 ml) and water (10 ml), sodium hydroxide (1.46 g, 36.4 mmol) was added, and the reaction solution was stirred at room temperature for 1 hour to complete the reaction. The reaction solution was concentrated, the resulting residue was dissolved in water (60 ml), 2 m chloristic acid was added to bring the value of the mixture of the mixture to 3-4, the mixture was extracted using DHM (50 ml × 3), the united organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated with reduced pressure with the receipt of the compound with the receipt of the compound with the receipt of the compound -3 (2.29 g, yellow oily liquid) with an output of 70%. MS m/h (ESI): 181.2 [M+1].

[0238] In step 4: compound 3-3 (2.29 g, 12.7 mmol) was dissolved in dry THF (80 ml), the reaction solution was cooled to -70°C under nitrogen gas protection, and a 2.5 M solution of butyllithium in THF (12.7 ml, 31.8 mmol) was slowly added dropwise. After completion of the dropwise addition, the reaction solution was slowly warmed to -40° C. and stirred for 45 minutes. Water (10 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (PE/EA=6/1) to give compound 3-4 (220 mg, yellow oily liquid) in 10.7% yield. MS m/h (ESI): 163.2 [M+1].

[0239] На шаге 5: соединение 3-4 (50 мг, 0,31 ммоль) и соединение 1а (80 мг, 0,31 ммоль) растворяли в ДХЭ (10 мл), добавляли изопропоксид титана (1 мл) и реакционный раствор перемешивают при 45°С в течение 18 ч. После охлаждения реакционного раствора до комнатной температуры добавляли боргидрид натрия (34 мг, 0,9 ммоль), реакционный раствор перемешивали в течение 3 ч, добавляли воду (5 мл), смесь перемешивали в течение 0,5 мин., фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной хроматографии (препаративная колонка: колонка С18 21,2 × 250 мм; система: 10 мМ NH ₄ HCO ₃ H ₂ O; длины волн: 254/214 нм; градиенты: 30%-60% изменений в ацетонитриле) с получением продукта H-3 (41,3 мг, белое твердое вещество) с выходом 32,6%. MS m/s (ESI): 407.3 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,51 (м, 1H), 7,70 (м, 1H), 7,44 (дд, J=8,1, 1,6 Гц, 1H), 7,28 (дд, J=5,5, 1,9 Гц, 1H), 7,17 (дд, J=7,4, 4,8 Гц, 1H), 5,75-5,59 (м, 1H), 3,95-3,67 (м, 2H), 3,66-3,41 (м, 3H), 2,77 (м, 1H), 2,44-2,37 (м, 1H), 2,35-2,27 (м, 1H), 2,02-1,70 (м, 5H), 1,66-1,25 (м, 8H), 0,97 (м, 2H), 0,65-0,34 (м, 4H), 0,27 (м, 1H).

[0240] Example 4.

6-ethyl-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (mixture of diastereomers H-4-1 and mixture of diastereomers H-4-2)

[0241] In step 1: propionaldehyde (2.9 g, 0.05 mol) was dissolved in acetonitrile (100 ml), triethylphosphonoacetate (11.8 g, 0.0525 mol) and lithium chloride (2.2 g, 0.0525 mol) were added, the reaction solution was cooled from 2 to 5°C, DBU (7.5 g, 0.0525 mol) was added slowly mol) and the reaction solution was kept at room temperature for 18 hours. The reaction solution was concentrated, and the resulting residue was diluted with EA (120 ml), then washed with water (80 ml × 1), washed with saturated saline (40 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE/EA=10/1) to give compound 4-1 (5.12 g, yellow oily liquid) in 80% yield. MS m/h (ESI): 129.2 [M+1].

[0242] In step 2: a solution of acetonitrile (90 ml) and DBU (8.52 g, 60 mmol) was added to a sealed tube containing compound 4-1 (5.12 g, 40 mmol) and pyrazole (4 g, 60 mmol), the reaction solution was stirred at 120 ° C for 18 hours, cooled to room temperature, concentrated and the resulting residue was diluted EA (100 ml), then washed with water (60 ml × 1), washed with brine (30 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE/EA=6/1) to give compound 4-2 (4.7 g, yellow oil) in 60% yield. MS m/h (ESI): 197.3 [M+1].

[0243] In step 3: Compound 4-2 (4.7 g, 24 mmol) was dissolved in a mixture of methanol (50 ml) and water (10 ml), sodium hydroxide (1.92 g, 48 mmol) was added, and the reaction solution was stirred at room temperature for 1 hour to complete the reaction. The reaction solution was concentrated, the resulting residue was dissolved in water (60 ml), 2 m chloristoric acid was added to bring the value of the mixture of the mixture to 3-4, the mixture was extracted using DHM (50 ml × 3), the united organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated with reduced pressure with the receipt of the compound with the receipt of the compound with the receipt of the compound -3 (2.62 g, yellow oily liquid) with an output of 65%. MS m/h (ESI): 169.2 [M+1].

[0244] In step 4: compound 4-3 (2.62 g, 15.6 mmol) was dissolved in dry THF (80 ml), the reaction solution was cooled to -70°C under nitrogen gas protection, and a 2.5 M solution of butyllithium in THF (15.6 ml, 39 mmol) was added slowly dropwise. After completion of the dropwise addition, the reaction solution was slowly warmed to -40° C. and stirred for 45 minutes. Water (10 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (PE/EA=6/1) to give compound 4-4 (350 mg, yellow oily liquid) in 15% yield. MS m/s (ESI): 151.2 [M+1].

[0245] In step 5: compound 3-4 (50 mg, 0.33 mmol) and compound 1a (87 mg, 0.33 mmol) were dissolved in DCE (10 ml), titanium isopropoxide (1 ml) was added, and the reaction solution was stirred at 45° C. for 18 hours. over 3 hours, water (5 ml) was added, the mixture was stirred for 0.5 minutes, filtered, and the resulting filtrate was concentrated under reduced pressure to give compound H-4 .

[0246] In step 6: compound H-4 was purified by preparative chromatography (preparative column: C18 column 21.2 x 250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to obtain a mixture of H-4-1 diastereomers (15.12 mg, white solid) in 11.6% yield: MS m/h (ESI): 395.2 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,54-8,47 (м, 1H), 7,71-7,68 (м, 1H), 7,44 (д, J=8,1 Гц, 1H), 7,29-7,27 (м, 1H), 7,21-7,13 (м, 1H), 6,03 (с, 1H), 5,71-5,63(м, 1H), 3,96-3,93 (м, 1H), 3,87-3,84 (м, 1H), 3,63-3,50 (м, 2H), 2,75-2,72 (м, 1H), 2,43-2,27 (м, 3H), 2,00-1,71 (м, 4H), 1,70-1,21 (м, 10H), 0,99-0,82 (м, 4H), 0,60-0,58 (м, 1H) и смесь диастереомеров H-4-2 (1,0 мг, белое твердое вещество) с выходом 0,8%: МС м/з (ИЭР): 395,2 [M+1].

[0247] Example 5.

6-cyclopentyl-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-5)

[0248] In step 1: cyclopentanecarboxaldehyde (4.9 g, 50 mmol) was dissolved in acetonitrile (100 ml), triethylphosphonoacetate (11.8 g, 52.5 mmol) and lithium chloride (2.2 g, 52.5 mmol) were added, the reaction solution was cooled from 2 to 5 ° C, DBU (7.5 g, 52.5 mmol) was added slowly and the reaction The solution was kept at room temperature for 18 hours. The reaction solution was concentrated, the resulting residue was diluted with EA (120 ml), then washed with water (70 ml × 1), washed with brine (40 ml × 1), dried over anhydrous sodium sulfate, and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE/EA=10/1) to give compound 5-1 (6.3 g, yellow oil) in 75% yield. MS m/h (ESI): 169.2 [M+1].

[0249] In step 2: a solution of acetonitrile (90 ml) and DBU (8.59 g, 56.5 mmol) was added to a sealed tube containing compound 5-1 (6.3 g, 37.5 mmol) and pyrazole (3.8 g, 56.2 mmol), the reaction solution was stirred at 100 ° C for 18 hours, cooled to room temperature, concentrated and the resulting residue was diluted with EA (100 ml), then washed with water (60 ml×1), washed with brine (30 ml×1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE/EA=6/1) to give compound 5-2 (4.86 g, yellow oil) in 55% yield. MS m/h (ESI): 237.3 [M+1].

[0250] In step 3: compound 5-2 (4.86 g, 20.6 mmol) was dissolved in a mixture of methanol (60 ml) and water (15 ml), sodium hydroxide (1.65 g, 41.25 mmol) was added, and the reaction solution was stirred at room temperature for 1 hour until the reaction was completed. The reaction solution was concentrated, the resulting residue was dissolved in water (40 ml), 2 m chloristoric acid was added to bring the mixture of the mixture from 3 to 4, the mixture was extracted using DHM (60 ml × 3), the united organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated with reduced pressure with the receipt of the compound with the receipt of the compound with the receipt of the compound -3 (3.2 g, yellow oily liquid) with an output of 75%. MS m/s (ESI): 209.2 [M+1].

[0251] In step 4: compound 5-3 (3.2 g, 15.45 mmol) was dissolved in dry THF (80 ml), the reaction solution was cooled to -70°C under the protection of nitrogen gas, and a 2.5 M solution of butyllithium in THF (15.45 ml, 38.62 mmol) was slowly added dropwise. After completion of the dropwise addition, the reaction solution was slowly warmed to -40° C. and stirred for 45 minutes. Water (10 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (PE/EA=6/1) to give compound 5-4 (293 mg, yellow oily liquid) in 10% yield. MS m/h (ESI): 191.3 [M+1].

[0252] На шаге 5: соединение 5-4 (57 мг, 0,30 ммоль) и соединение 1а (78 мг, 0,30 ммоль) растворяли в ДХЭ (10 мл), добавляли изопропоксид титана (1 мл) и реакционный раствор перемешивали при 45°С в течение 18 ч. После охлаждения реакционного раствора до комнатной температуры добавляли боргидрид натрия (34 мг, 0,9 ммоль), реакционный раствор перемешивали в течение 3 ч., добавляли воду (5 мл), смесь перемешивали в течение 0,5 мин., фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной хроматографии (препаративная колонка: колонка С18 21,2 × 250 мм; система: 10 мМ NH ₄ HCO ₃ H ₂ O; длины волн: 254/214 нм; градиенты: 30%-60% изменений в ацетонитриле) с получением соединения H-5 (33 мг, бесцветное маслянистое вещество) с выходом 25,4%. MS m/s (ESI): 435.1 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,50 (дд, J=4,6, 2,1 Гц, 1H), 7,70-7,67 (м, 1H), 7,43 (д, J=8,1 Гц, 1H), 7,27 (дд, J=4,0, 1,8 Гц, 1H), 7,17 (дд, J=7,4, 4,8 Гц, 1H), 5,67 (дд, J=24,4, 1,8 Гц, 1H), 3,96-3,93 (м, 1H), 3,85-3,81 (м, 1H), 3,58-3,55 (м, 2H), 2,76-2,63 (м, 1H), 2,44-2,26 (м, 3H), 2,13-2,09 (м, 1H), 1,99-1,18 (м, 21H), 0,94 (с, 1H), 0,60-0,56 (м, 1H).

[0253] Example 6.

6-tert-Butyl-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-6)

[0254] In step 1: pivalaldehyde (17.2 g, 0.2 mmol) was dissolved in acetonitrile (400 ml), triethylphosphonoacetate (48 g, 0.21 mmol) and lithium chloride (8.8 g, 0.21 mmol) were added, the reaction solution was cooled from 2 to 5 ° C, DBU (30.4 g, 0.2 mol) was added slowly and the reaction solution was kept at room temperature for 18 hours. The reaction solution was concentrated, and the resulting residue was diluted with EA (300 ml), then washed with water (100 ml × 1), washed with brine (80 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE/EA=10/1) to give compound 6-1 (25.6 g, yellow oil) in 82% yield. MS m/h (ESI): 157.2 [M+1].

[0255] In step 2: a solution of acetonitrile (80 ml) and DBU (11.4 g, 75 mmol) was added to a sealed tube containing compound 6-1 (7.8 g, 50 mmol) and pyrazole (5.1 g, 75 mmol), the reaction solution was stirred at 150 ° C for 48 hours, cooled to room temperature, concentrated, the resulting residue was diluted EA (100 ml) was poured, then washed with water (60 ml × 1), washed with brine (30 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE/EA=6/1) to give compound 6-2 (8.96 g, yellow oil) in 80% yield. MS m/s (ESI): 225.1 [M+1].

[0256] In step 3: compound 6-2 (8.96 g, 40 mmol) was dissolved in a mixture of methanol (100 ml) and water (20 ml), sodium hydroxide (3.2 g, 80 mmol) was added and the reaction solution was stirred at room temperature for 1 hour until the reaction was completed. The reaction solution was concentrated, the resulting residue was dissolved in water (70 ml), 2 M hydrochloric acid was added to adjust the pH of the mixture from 3 to 4, the mixture was extracted with DCM (90 ml × 3), the combined organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure to obtain compound 6-3 (7 g, yellow oily liquid) with a yield of 90%. MS m/h (ESI): 197.2 [M+1].

[0257] In step 4: compound 6-3 (7 g, 35.7 mmol) was dissolved in dry THF (120 ml), the reaction solution was cooled to -70°C under the protection of nitrogen gas, and a 2.5 M solution of butyllithium in THF (35.7 ml, 89.2 mmol) was slowly added dropwise. After completion of the dropwise addition, the reaction solution was slowly warmed to -40° C. and stirred for 45 minutes. Water (3 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (PE/EA=6/1) to give compound 6-4 (4.76 g, yellow oily liquid) in 15% yield. MS m/h (ESI): 179.2 [M+1].

[0258] In step 5: compound 6-4 (25 mg, 0.14 mmol) was dissolved in DCE (5 ml), compound 1a (36 mg, 0.14 mmol) and titanium isopropoxide (0.5 ml) were added, and the reaction solution was stirred at 60° C. for 12 hours. Sodium borohydride (26.5 mg, 0.7 mmol) was added and the reaction solution was stirred at 65° C for 3 hours. Water (20 ml) was added, the mixture was filtered and the resulting filtrate was extracted with DCM (40 ml×2), then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting concentrate was purified by preparative chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give compound H-6 (15.21 mg, white liquid) in 25.74 %. MS m/h (ESI): 423.2 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,55-8,46 (м, 1H), 7,81-7,69 (м, 1H), 7,50 (д, J=8,0 Гц, 1H), 7,37 (дд, J=3,9, 1,9 Гц, 1H), 7,29-7,18 (м, 1H), 5,83 (дд, J=20,9, 1,2 Гц, 1H), 3,94 (ддд, J=16,0, 12,3, 4,8 Гц, 2H), 3,82-3,68 (м, 2H), 2,72-2,58 (м, 2H), 2,52 (д, J=14,5 Гц, 1H), 2,41 (дд, J=13,8, 2,0 Гц, 1H), 2,14-1,99 (м, 2H), 1,95-1,85 (м, 2H), 1,80-1,36 (м, 8H), 1,09 (с, 1H), 1,00 (д, J=1,8 Гц, 9H), 0,77-0,67 (м, 1H).

[0259] Example 7.

6-(2-fluoropropan-2-yl)-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-7)

[0260] In step 1: 2-fluoro-2-methylpropan-1-ol (1 g, 10.86 mmol) was dissolved in a mixture of acetonitrile (15 ml) and DCM (3 ml), (diacetoxyiodo)benzene (3.67 g, 11.39 mmol) and 2,2,6,6-tetramethylpiperidinooxy (85 mg, 0.54 mmol) was added and the mixture reacted at room temperature overnight to give compound 7-1 (978 mg).

[0261] In step 2: lithium chloride (690 mg, 16.28 mmol) was added to acetonitrile (30 ml), triethylphosphonoacetate (2.92 g, 13.02 mmol) and DBU (5.3 g, 34.81 mmol) were successively added after the reaction solution was cooled to 0°C, compound 7-1 (978 mg) was added dropwise and the reaction the solution was reacted at room temperature overnight. Water (50 ml) was added, the mixture was extracted with EA (50 ml × 3), the resulting organic phases were combined, the combined organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluents (PE/EA=1/010/1) to give compound 7-2 (734 mg, colorless oily liquid ) with a yield of 42%. MS m/h (ESI): 161.1 [M+1].

[0262] In Step 3: Compound 7-2 (734 mg, 4.58 mmol), pyrazole (468 mg, 6.87 mmol) and potassium carbonate (1.27 g, 9.19 mmol) were added to DMF (20 ml), and the reaction solution was reacted at 65° C. for 18 hours. After the reaction solution was cooled to room temperature, EA (300 ml) was added, followed by washing with water ( 30 ml) and saturated sodium chloride solution (30 ml × 2). The resulting organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluents (PE/EA=1/010/1) to give compound 7-3 (1 g, colorless oily liquid) in 96% yield. MS m/s (ESI): 229.1 [M+1].

[0263] На шаге 4: соединение 7-3 (1 г, 4,38 ммоль) растворяли в смеси ТГФ (20 мл) и метанола (5 мл), добавляли моногидрат гидроксида лития (368 мг, 8,77 ммоль) и воды (5 мл) и реакционный раствор перемешивали при комнатной температуре в течение 1 ч. Добавляли 1 М раствор хлористоводородной кислоты для доведения значения рН реакционного раствора до 3, органический растворитель удаляли центрифугированием при пониженном давлении, полученную водную фазу экстрагировали с помощью ДХМ (30 мл × 4), полученные органические фазы объединяли, объединенную органическую фазу сушили над безводным сульфатом натрия и фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной жидкостной хроматографии с получением соединения 7-4 (588 мг, белое твердое вещество) с выходом 67%. MS m/s (ESI): 201.0 [M+1].

[0264] На шаге 5: соединение 7-4 (588 мг, 2,94 ммоль) растворяли в ТГФ (15 мл), смесь охлаждали до -78°С, медленно по каплям добавляли 2,5 М н-бутиллития (2,9 мл, 7,25 ммоль), реакционный раствор реагировал при -60°С в течение 1 ч., а затем реагировал при -20°С в течение 1 ч. Для гашения реакции добавляли воду (15 мл), смесь экстрагировали ЭА (30 мл × 3), полученные органические фазы объединяли, объединенную органическую фазу сушили над безводным сульфатом натрия и фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью тонкослойной хроматографии с использованием проявляющей системы растворителей (ПЭ/ЭA=3/1) с получением соединения 7-5 (39 мг, бледно-желтое твердое вещество) с выходом 7,3%. MS m/s (ESI): 183.0 [M+1].

[0265] In step 6: compound 7-5 (39 mg, 0.214 mmol), compound 1a (67 mg, 0.257 mmol) and titanium isopropoxide (1 ml) were dissolved in DCE (10 ml) and the reaction solution was reacted at 50° C. overnight. Water (1 ml) was added to quench the reaction, the mixture was stirred at room temperature for 20 minutes, filtered, and the resulting filtrate was concentrated under reduced pressure. The resulting residue was dissolved in methanol (10 ml), sodium borohydride (30 mg, 0.793 mmol) was added, and the reaction solution was reacted at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% change in acetonitrile) to give compound H-7 (35.28 mg, colorless oily liquid) in 39% yield. MS m/s (ESI): 427.1 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,55-8,47 (м, 1H), 7,78-7,73 (м, 1H), 7,50 (д, J=8,0 Гц, 1H), 7,42 (дд, J=5,6, 1,7 Гц, 1H), 7,23 (дд, J=6,6, 5,1 Гц, 1H), 5,86 (дд, J=18.4, 1,3 Гц, 1H), 4,36-4,22 (м, 1H), 4,05-3,94 (м, 1H), 3,82-3,66 (м, 2H), 2,95-2,82 (м, 1H), 2,65-2,37 (м, 3H), 2,18-1,85 (м, 4H), 1,82-1,18 (м, 14H), 1,12-1,06 (м, 1H), 0,75-0,67 (м, 1H).

[0266] Example 8.

6-isobutyl-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-8)

[0267] In step 1: lithium chloride (3.69 g, 87.05 mmol) was added to acetonitrile (40 ml), then triethylphosphonoacetate (19.5 g, 86.98 mmol) and DBU (10.6 g, 69.63 mmol) were added successively, after the reaction solution was cooled to 0°C, 3-methylbutyraldehyde (5 g, 58.08 mmol) and the reaction solution was reacted at room temperature overnight. Saturated sodium chloride solution (150 ml) was added, the mixture was extracted with EA (50 ml × 3), the resulting organic phases were combined, the combined organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluents (PE/EA=1/0-10/1) to give compound 8-1 (6.3 g , colorless oily liquid) with a yield of 70%. MS m/h (ESI): 157.0 [M+1].

[0268] На шаге 2: к ДМФ (20 мл) добавляли соединение 8-1 (1 г, 6,40 ммоль), пиразол (870 мг, 12,78 ммоль), карбонат калия (1,8 г, 13,02 ммоль) и реакционный раствор реагировал при 65°С в течение 18 ч. После охлаждения реакционного раствора до комнатной температуры добавляли ЭА (200 мл), смесь промывали насыщенным раствором хлорида натрия (50 мл × 3), объединенную органическую фазу сушили над безводным сульфатом натрия, фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью колоночной хроматографии на силикагеле с использованием элюентов (ПЭ/ЭA=от 1/0 до 4/1) с получением соединения 8-2 (1,2 г, бесцветная маслянистая жидкость) с выходом 84%. MS m/s (ESI): 225.1 [M+1].

[0269] На шаге 3: соединение 8-2 (1,2 г, 5,35 ммоль) растворяли в смеси ТГФ (20 мл) и метанола (5 мл), добавляли гидроксид натрия (428 мг, 10,7 ммоль) и воду (5 мл) и реакционный раствор перемешивали при комнатной температуре в течение 2 ч. Добавляли 6 М раствор хлористоводородной кислоты для доведения значения рН реакционного раствора до 4, органический растворитель удаляли центрифугированием при пониженном давлении, полученную водную фазу экстрагировали с помощью ДХМ (50 мл × 3), полученные органические фазы объединяли, объединенную органическую фазу сушили над безводным сульфатом натрия и фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной жидкостной хроматографии с получением соединения 8-3 (1 г, белое твердое вещество) с выходом 96%. MS m/h (ESI): 197.1 [M+1].

[0270] In step 4: compound 8-3 (1 g, 5.09 mmol) was dissolved in THF (40 ml), the mixture was cooled to -78°C, 2.5 M n-butyllithium (4.7 ml, 11.75 mmol) was slowly added dropwise, and the reaction solution was reacted at -78°C for 1 h. To quench the reaction, a saturated solution of ammonium chloride (1 ml), the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using a developing solvent system (PE/EA=3/1) to obtain compound 8-4 (57 mg, colorless oily liquid) with a yield of 6.3%. MS m/h (ESI): 179.1 [M+1].

[0271] In step 5: compound 8-4 (57 mg, 0.32 mmol), compound 1a (100 mg, 0.38 mmol) and titanium isopropoxide (1 ml) were dissolved in DCE (15 ml) and the reaction solution was reacted at 50° C. overnight. Water (1.5 ml) was added to quench the reaction, the mixture was stirred at room temperature for 20 minutes, filtered, and the resulting filtrate was concentrated under reduced pressure. The resulting residue was dissolved in methanol (10 ml), sodium borohydride (30 mg, 0.793 mmol) was added, and the reaction solution was reacted at room temperature for 10 minutes. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give product H-8 (25.26 mg, pale yellow oily liquid) in 19% yield. MS m/h (ESI): 423.3 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,52-8,51 (м, 1H), 7,78-7,74 (м, 1H), 7,50 (д, J=8,1 Гц, 1H), 7,38 (дд, J=6,1, 1,7 Гц, 1H), 7,25-7,21 (м, 1H), 5,84 (дд, J=26,2, 1,3 Гц, 1H), 4,17-4,04 (м, 2H), 3,81-3,66 (м, 2H), 2,98-2,90 (м, 1H), 2,65-2,47 (м, 2H), 2,41 (д, J=13,9 Гц, 1H), 2,15-1,88 (м, 4H), 1,83-1,35 (м, 11H), 1,12-1,06 (м, 1H), 0,94 (дд, J=14,5, 6,7 Гц, 6H), 0,76-0,67 (м, 1H).

[0272] Example 9.

3-fluoro-6-isopropyl-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-9)

[0273] In step 1: 6-isopropyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-one (compound 11-4, 250 mg, 1.52 mmol) was dissolved in methanol (15 ml), sodium borohydride (115 mg, 3.04 mmol) was added in portions, and the reaction solution was reacted at room temperature for 1 hour. saturated ammonium chloride solution (1 ml), the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using the eluent system (DCM/methanol = 1/0 to 10/1) to obtain compound 9-1 (249 mg, yellow oily substance) in 98% yield. MS m/h (ESI): 167.1 [M+1].

[0274] In Step 2: Compound 9-1 (249 mg, 1.50 mmol) was dissolved in acetonitrile (10 mL), 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (1.1 g, 3.19 mmol) was added and the reaction solution was stirred at room temperature overnight. Water (40 ml) was added, the mixture was extracted with EA (40 ml × 3), the resulting organic phases were combined, the combined organic phase was washed with saturated sodium chloride solution (50 ml), dried over anhydrous sodium sulfate, and filtered, and the resulting filtrate was concentrated under reduced pressure to obtain compound 9-2 (275 mg, yellow oily substance) with a yield of 99%. MS m/z (ESI): 185.1 [M+1].

[0275] In step 3: compound 9-2 (275 mg) was dissolved in DCM (20 ml), Dess-Martin periodinan (1.27 g, 2.99 mmol) was added, and the reaction solution was reacted at room temperature for 2 hours. Saturated sodium bicarbonate solution (2 ml) was added to quench the reaction, the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified using a preparative thin layer chromatography using a chromatographic system (PE/EA=3/1) to give compound 9-3 (110 mg, colorless oil) in 40% yield. MS m/z (ESI): 183.1 [M+1].

[0276] In step 4: compound 9-3 (56 mg, 0.31 mmol), compound 1a (88 mg, 0.34 mmol) and titanium isopropoxide (1 ml) were dissolved in DCE (12 ml). The reaction solution was reacted at 50°C overnight, sodium borohydride (80 mg, 2.11 mmol) was added, and the reaction solution was reacted at 60°C for 2 h. 250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% change in acetonitrile) to give compound H-9 (14.92 mg, white oil) in 25% yield. MS m/s (ESI): 427.1 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,54-8,45 (м, 1H), 7,76-7,71 (м, 1H), 7,48 (д, J=8,1 Гц, 1H), 7,33-7,14 (м, 2H), 4,17-3,98 (м, 2H), 3,83-3,65 (м, 2H), 2,74-2,47 (м, 3H), 2,44-2,29 (м, 2H), 2,26-1,84 (м, 4H), 1,81-1,29 (м, 8H), 1,12-1,06 (м, 1H), 1,02-0,86 (м, 3H), 0,81- 0,54 (м, 4H).

[0277] Example 10.

6-(1-methylcyclopropyl)-N-(2-(( R )-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-10)

[0278] In step 1: 1-methylcyclopropanecarboxylic acid (10 mg, 0.10 mol) and N,O-dimethylhydroxylamine hydrochloride (14.6 g, 0.15 mol) were added to DMF (80 ml), HATU (45.6 g, 0.12 mol) and DIEA (50 ml, 0.30 ml) were added successively, and the reaction solution was kept at room temperature during the night. EA (500 ml) was added. The resulting organic phase was washed with 1 N. hydrochloric acid (100 ml × 2) and saturated sodium chloride solution (50 ml × 2), dried over anhydrous sodium sulfate, filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using an eluent system (PE/EA=1/0 to 1/1) to obtain compound 10-1 (11.3 g, pale -yellow oily substance) with a yield of 79%. MS m/z (ESI): 144.1 [M+1].

[0279] In step 2: compound 10-1 (14.5 g, 101 mmol) was dissolved in diethyl ether (90 ml), and after cooling the mixture to -78°C, 1 M diisobutylaluminum hydride (152 ml, 152 mmol) was added slowly dropwise. The reaction solution was reacted at -78° C. for 2 hours, 1 M HCl (16 ml) was added to quench the reaction, and after the reaction mixture was heated to 0° C., 1 M HCl (16 ml) was added. The resulting mixture was dried over anhydrous sodium sulfate (70 g) and filtered to give compound 10-2 .

[0280] In step 3: lithium chloride (6.5 g, 153 mmol) was added to acetonitrile (60 ml), triethylphosphonoacetate (24.5 ml, 122 mmol) and DBU (18.1 ml, 121 mmol) were added successively after the reaction solution was cooled to 0°C, compound 10-2 (8.5 g, 101 mmol) was added dropwise and the reaction solution was reacted at room temperature overnight. Water (150 ml) was added, the mixture was extracted with EA (150 ml × 3), the resulting organic phases were combined, the combined organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluents (PE/EA=1/010/1) to give compound 10-3 (10.5 g, colorless oily liquid) with a yield of 68%. MS m/s (ESI): 155.0 [M+1].

[0281] In Step 4: Compound 10-3 (10.5 mg, 0.068 mmol), pyrazole (9.3 mg, 0.137 mmol) and potassium carbonate (18.8 g, 0.136 mmol) were added to DMF (50 ml), and the reaction solution was reacted at 65°C for 45 hours. After the reaction solution was cooled to room temperature, EA (300 ml) was added, followed by washing with water (100 ml) and saturated sodium chloride solution (100 ml × 2). The resulting organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluents (PE/EA=1/0 to 5/1) to give compound 10-4 (12.3 g, colorless oily liquid) in 82% yield. MS m/s (ESI): 223.1 [M+1].

[0282] На шаге 5: соединение 10-4 (12,3 г, 55,3 ммоль) растворяли в смеси ТГФ (50 мл) и метанола (20 мл), добавляли моногидрат гидроксида лития (3,6 г, 85,7 ммоль) и воды (30 мл) и реакционный раствор перемешивали при комнатной температуре в течение 2 ч. Добавляли 6 М раствор хлористоводородной кислоты для доведения значения рН реакционного раствора до 3, органический растворитель удаляли центрифугированием при пониженном давлении, полученную водную фазу экстрагировали с помощью ДХМ (100 мл × 3), полученные органические фазы объединяли, объединенную фазу сушили над безводным сульфатом натрия и фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной жидкостной хроматографии с получением соединения 10-5 (9,2 г, бесцветная маслянистая жидкость) с выходом 86%. MS m/s (ESI): 195.0 [M+1].

[0283] На шаге 6: соединение 10-5 (3,05 г, 15,7 ммоль) растворяли в ТГФ (80 мл), после смесь охлаждали до -78°C, медленно по каплям добавляли 2,5 М н-бутиллитий (15,7 мл, 39,3 ммоль) и реакционный раствор вступал в реакцию при -78°C в течение 0,5 ч. и вступал в реакцию при -45°C в течение 1 ч. Для гашения реакции добавляли насыщенный раствор хлорида аммония (30 мл), смесь экстрагировали с помощью ЭА (50 мл × 4), полученные органические фазы объединяли, объединенную органическую фазу сушили над безводным сульфатом натрия и фильтровали, полученный фильтрат концентрировали при пониженном давлении, а полученный остаток очищали с помощью колоночной хроматографии на силикагеле с использованием элюентов (ПЭ/ЭА=от 1/0 до 3/1) с получением соединения 10-6 (460 мг, бесцветная маслянистая жидкость) с выходом 16,7%. MS m/h (ESI): 177.0 [M+1].

[0284] In step 7: compound 10-6 (58 mg, 0.33 mmol), compound 1a (86 mg, 0.33 mmol) and titanium isopropoxide (1 ml) were dissolved in DCE (10 ml) and the reaction solution was reacted at 50°C overnight. Sodium borohydride (80 mg, 2.1 mmol) was added and the reaction solution was reacted at 60° C. for 2 hours. Water (1 ml) was added to quench the reaction, the mixture was stirred at room temperature for 30 minutes, filtered, and the resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give compound H-10 (51.14 mg, colorless oily liquid) with 37% yield. MS m/s (ESI): 421.1 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,55-8,47 (м, 1H), 7,80-7,71 (м, 1H), 7,51 (д, J=8,1 Гц, 1H), 7,45-7,38 (м, 1H), 7,27-7,19 (м, 1H), 5,86 (дд, J=21,6, 1,9 Гц, 1H), 4,02 (т, J=7,7 Гц, 1H), 3,80-3,73 (м, 2H), 3,49 (т, J=7,1 Гц, 1H), 2,81-2,72 (м, 1H), 2,68-2,49 (м, 2H), 2,41 (дд, J=13,9, 2,0 Гц, 1H), 2,17-1,96 (м, 3H), 1,91 (д, J=13,9 Гц, 1H), 1,78-1,36 (м, 8H), 1,12-1,07 (м, 1H), 0,83 (д, J=5,3 Гц, 3H), 0,77-0,66 (м, 2H), 0,60-0,54 (м, 1H), 0,50-0,43 (м, 1H), 0,37-0,29 (м, 1H).

[0285] Example 11.

6-(isopropyl)-N-(2-(( R )-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-11)

[0286] In Step 1: Triethylphosphonoacetate (476 ml, 2.4 mol), DBU (365 g, 2.4 mol), lithium chloride (127 g, 3 mol) and acetonitrile (1.2 L) were added to a 3 L single-necked flask, and the reaction solution was stirred at room temperature for 20 minutes. protected by argon gas. After the reaction solution was cooled to 0° C. (internal temperature), isobutyraldehyde (144 g, 2 mol) was slowly added dropwise. The reaction solution was stirred at room temperature for 12 hours. After completion of the reaction process, controlled by LC-MS, the reaction solution was filtered and the resulting filter cake was washed with EA (100 ml × 2). Water (1 L) was added, the mixture was extracted with EA (1.5 L×2), the combined organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and dried by centrifugation to give compound 11-1 (175 g, colorless liquid).

[0287] In step 2: DMF (1 L) was added to a one-necked flask (3 L) into which compound 11-1 (300 g, 2.1 mol) was placed, potassium carbonate (579 g, 4.2 mol) and pyrazole (287 g, 4.2 mol) were added with stirring, the reaction solution was stirred at 65° C. for 18 hours, after completion of the reaction process controlled LC-MS, the reaction solution was dried directly by centrifugation and the resulting solid was dissolved in acetonitrile (150 ml) to obtain a suspension, and the suspension was filtered to obtain a white solid. The white solid was filtered off and the resulting filter cake was washed with EA (500 ml x 2), then washed with brine (300 ml x 3), dried over anhydrous sodium sulfate, dried by centrifugation and purified by column chromatography (mobile phase - PE containing 5% EA) to give compound 11-2 (258 g, colorless liquid) with yield ohm 58%. MS m/z (ESI): 211.1 [M+1].

[0288] In step 3: potassium hydroxide (133 g, 3.32 mol) was dissolved in 200 ml of water and the resulting aqueous potassium hydroxide solution was pre-cooled to 5°C. Compound 11-2 (465 g, 2.21 mol), methanol (0.5 L) and THF (0.5 L) were added to a flask (3 L), pre-cooled aqueous potassium hydroxide solution was added, the reaction solution was stirred for 2 hours, concentrated hydrochloric acid was added to adjust the pH of the reaction solution to about 3, the mixture was extracted with DCM (800 ml × 2), all obtained the organic phases were combined, the combined organic phase was washed with brine, dried and concentrated to give 11-3 (410.5 g, white solid) in 100% yield. MS m/z (ESI): 183.1 [M+1].

[0289] In step 4: compound 11-3 (10.2 g, 0.055 mol) and THF (200 ml) were placed in a three-necked flask (500 ml) under nitrogen gas protection and the mixture was cooled to -75°C. A 2.5 M solution of n-butyllithium in THF (55 ml, 0.137 mol) was added slowly dropwise and the dropwise addition was completed, the reaction solution was slowly heated to -10°C and stirred for 3 h. the phase was washed with brine, dried and concentrated to give a brown liquid. The brown liquid was purified by column chromatography (mobile phase was PE containing 30% EA) to give compound 11-4 (4 g, white solid) in 22.2% yield. MS m/z (ESI): 165.1 [M+1].

[0290] In step 5: compound 11-4 (50 mg, 0.3 mmol) was dissolved in DCE (5 ml), compound 1a (79 mg, 0.3 mmol) and titanium isopropoxide (0.5 ml) were added, and the reaction solution was stirred at 45°C for 16 h. Sodium borohydride (39 mg, 1 mmol) was added. The reaction solution was stirred at 45° C. for 3 hours. Water (20 ml) was added, the mixture was filtered, and the resulting filtrate was extracted with DCM (20 ml×2), then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting concentrate was purified by preparative chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give compound H-11 (2.9 mg, white solid) in a yield of 2.37 %. MS m/s (ESI): 409.2 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,54 (д, 1H), 7,73 (т, 1H), 7,47 (д, 1H), 7,34-7,32 (м, 1H), 7,22-7,18 (м, 1H), 5,71 (д, 1H), 4,00-3,88 (м, 2H), 3,61-3,55 (м, 2H), 2,62-2,53 (м, 1H), 2,48-2,43 (м, 3H), 2,39-2,30 (м, 2H), 2,05-1,64 (м, 7H), 1,63-1,32 (м, 6H), 0,95 (д, 3H), 0,73 (д, 3H), 0,64-0,51(м, 1H).

[0291] Example 12.

6-methyl-N-(2-(( R )-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-12-1 diastereomer mixture and H-12-2 diastereomer mixture)

[0292] In step 1: DMF (30 ml) was placed in a one-necked flask (100 ml), into which ethyl (E)-but-2-enoate (3.5 g, 30 mmol) was placed, potassium carbonate (4.14 g, 32 mmol) and pyrazole (2.73 g, 32 mmol) were added with stirring, and the reaction solution was stirred at 50 ° C for 20 hours until completion reaction process controlled by LC-MS. The reaction solution was filtered and the resulting filter cake was washed with EA (50 ml × 2), then washed with saturated brine (30 ml × 3), dried over anhydrous sodium sulfate, dried by centrifugation and purified by column chromatography (mobile phase - PE containing 5% EA) to obtain compound 12-1 (3.5 g, yellow liquid) with a yield of 6 4%. MS m/z (ESI): 183.1 [M+1].

[0293] In step 2: sodium hydroxide (0.8 g, 20 mmol) was dissolved in water (5 ml) and the resulting aqueous sodium hydroxide solution was pre-cooled to 5°C. Compound 12-1 (1.82 g, 10 mol), methanol (10 ml) and THF (10 ml) were placed in a flask (100 ml), a pre-cooled aqueous solution of potassium hydroxide was added, the reaction solution was stirred for 2 hours, concentrated hydrochloric acid was added to adjust the pH of the reaction solution to about 4, the mixture was extracted with DCM (100 ml × 2), all the resulting organic phases were combined was added and the combined organic phase was washed with brine, dried and concentrated to give 12-2 (1.2 g, white solid) in 78% yield. MS m/z (ESI): 155.1 [M+1].

[0294] In step 3: compound 12-2 (0.8 g, 5.19 mol) and THF (10 ml) were placed in a three-necked flask (100 ml) under the protection of nitrogen gas and the mixture was cooled to -75°C. A 2.5 M solution of n-butyllithium in THF (5.2 ml, 12.9 mol, 2.5 mol/l) was slowly added dropwise, and after completion of the dropwise addition, the reaction solution was stirred at -75°C for 3 h. The organic phase was washed with brine, dried and concentrated to give a brown liquid. The brown liquid was purified by column chromatography (mobile phase was PE containing 30% EA) to give compound 12-3 (45 mg, colorless liquid) in 6.4% yield. MS m/z (ESI): 137.1 [M+1].

[0295] In step 4: compound 12-3 (26 mg, 0.2 mmol) was dissolved in DCE (5 ml), compound 1a (52 mg, 0.2 mmol) and titanium isopropoxide (0.5 ml) were added and the reaction solution was stirred at 45°C for 6 hours. Sodium borohydride (38 mg, 1 mmol) was added and the reaction solution was stirred at 45°C for 3 hours Water (20 ml) was added, the mixture was filtered and the resulting filtrate was extracted with DCM (40 ml×2), then dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product H-12 .

[0296] At step 5: the resulting concentrate, i.e. the crude H-12 product was purified by preparative chromatography (prep column: C18 column 21.2 x 250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give a mixture of H-12-1 diastereomers (2.04 mg , white solid) in 2.68% yield: MS m/h (ESI): 381.3 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,56 (дд, J=3,9, 0,9 Гц, 1H), 8,44 (brs, 1H), 7,85-7,75 (м, 1H), 7,58-7,45 (м, 2H), 7,32-7,24 (м, 1H), 5,99 (дд, J=45,9, 1,7 Гц, 1H), 4,67-4,58 (м, 1H), 4,42 (т, J=7,4 Гц, 1H), 4,30 (дq, J=13,4, 6,7 Гц, 1H), 3,82-3,70 (м, 2H), 3,18-3,06 (м, 1H), 2,98-2,81 (м, 1H), 2,50 (дд, J=24,4, 13,6 Гц, 2H), 2,42-2,35 (м, 1H), 2,08 (ддд, J=25,9, 12,6, 4,4 Гц, 1H), 1,99-1,87 (м, 2H), 1,86-1,66 (м, 3H), 1,66-1,29 (м, 7H), 1,15-1,02 (м, 1H), 0,79-0,62 (м, 1H) и смесь диастереомеров H-12-2 (0,83 мг, белое твердое вещество) с выходом 1,09%: ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,59-8,54 (м, 1H), 8,36 (brs, 1H), 7,85-7,75 (м, 1H), 7,57-7,44 (м, 2H), 7,31-7,23 (м, 1H), 5,98 (дд, J=45,0, 1,4 Гц, 1H), 4,67-4,58 (м, 2H), 4,45-4,36 (м, 1H), 4,30 (дд, J=13,6, 6,8 Гц, 1H), 3,82-3,67 (м, 2H), 3,19-3,08 (м, 1H), 2,96-2,84 (м, 1H), 2,50 (дд, J=24,4, 13,5 Гц, 2H), 2,43-2,28 (м, 1H), 2,08 (ддд, J=17.2, 12,9, 4,8 Гц, 1H), 2,00-1,87 (м, 2H), 1,86-1,27 (м, 9H), 1,12-1,05 (м, 1H), 0,77-0,65 (м, 1H).

[0297] Example 13.

6-(1-methylcyclopropyl)-N-(2-(( R )-9-(4-fluorophenyl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-6-isopropyl-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole-4-amine (H-13)

[0298] In step 1: cyclopentanone (126 g, 1.5 mol) and 3-buten-1-ol (108 g, 2.4 mol) were placed in a one-necked flask, and after the reaction solution was cooled to 0°C (internal temperature), 70% sulfuric acid (500 ml) was slowly added dropwise. The reaction solution was stirred at room temperature for 12 hours until completion of the reaction, controlled by TLC (mobile phase was PE containing 20% EA). After the reaction solution was cooled to 0°C, ice water (1400 ml) was added, the mixture was washed and extracted with ethyl acetate (500 ml×2). The combined organic phase was washed with brine (100 ml×2), dried over anhydrous sodium sulfate, and dried by centrifugation, and the resulting residue was purified using column chromatography (mobile phase - DCM containing 10% methanol) to obtain compound 13-1 (142 g, brown liquid) with a yield of 60.9%.

[0299] In Step 2: Compound 13-1 (56 g, 0.35 mol), pyridinium chlorochromate (100.5 g, 0.47 mol), silica gel (220 g) and DCM (0.5 L) were placed in a one-necked flask (2 L) and the reaction solution was stirred at room temperature for 12 hours under argon. The reaction solution was filtered, the resulting filtrate was concentrated, and the resulting residue was purified by column chromatography (mobile phase was PE containing 20% EA) to obtain compound 13-2 (110 g, white oily liquid) in 91.5% yield. MS m/z (ESI): 155.1 [M+1].

[0300] In step 3: Toluene (0.35 L) was added to a single-necked flask (2 L) into which compound 13-2 (85 g, 0.55 mol) and methyl 2-cyanoacetate (65.6 g, 0.66 mol) were placed, ammonium acetate (12.8 g, 0.17 mol) and acetic acid (5.5 ml) were added with stirring. The reaction solution was stirred at 120°C for 5 hours to complete the reaction process, controlled by LC-MS. The reaction solution was cooled and stratified, the resulting aqueous phase was extracted with ethyl acetate (200 ml × 2), all the resulting organic phases were combined and the combined organic phase was washed with aqueous sodium bicarbonate solution, washed with saturated saline, dried and concentrated to obtain compound 13-3 (106 g, yellow liquid) with a yield of 81.7%. MS m/z (ESI): 236.1 [M+1].

[0301] In Step 4: THF (200 ml), 4-fluorophenylmagnesium bromide (72 ml, 0.14 mol, 2 mol/l) and copper iodide (2.3 g, 0.012 mol) were placed in a three-necked flask (500 ml) under protection with nitrogen gas, and the reaction solution was stirred for 0.5 h. A solution of compound 13- 3 (28.2 g, 0.12 mol) in THF (50 ml), and after completion of the dropwise addition, the reaction solution was slowly warmed to room temperature and stirred for 12 h. Ice water (500 ml) was added, the mixture was extracted with EA (600 ml × 2), the resulting organic phases were combined and the combined organic phase was washed with saturated saline, dried and concentrated to obtain a brown liquid. The brown liquid was purified by column chromatography (mobile phase was PE containing 10% EA) to give compound 13-4 (27 g, yellow solid) in 68% yield. MS m/z (ESI): 332.1 [M+1].

[0302] In step 5: compound 13-4 (12 g, 0.036 mol), potassium hydroxide (4.1 g, 0.072 mol) and ethylene glycol (84 ml) were placed in a three-necked flask (500 ml) and the reaction solution was stirred at 120°C for 5 hours. Ice water (300 ml) was added, the mixture was extracted with EA (500 ml × 2), the obtained the organic phases were combined and the combined organic phase was washed with brine, dried and concentrated to give a brown liquid. The brown liquid was purified by column chromatography (mobile phase was PE containing 20% EA) to give compound 13-5 (19.4 g, colorless liquid) in 82.6% yield.

[0303] Step 6: Performed a chiral resolution of compound 13-5 by supercritical liquid chromatography (SFC) to give (R)-2-(9-(4-fluorophenyl)-6-oxaspiro[4.5]decan-9-yl)acetonitrile (compound 13-6 , 8.51 g, colorless liquid) in 41% yield. MS m/s (ESI): 274.1 [M+1].

[0304] In step 7: Compound 13-6 (1 g, 3.6 mmol), ethanol (6 ml), ammonia water (0.6 ml) and Raney nickel (0.1 g) serving as a catalyst (100 ml) were placed in a one-necked flask, and the reaction solution was stirred for 16 h under the protection of hydrogen gas. The reaction solution was filtered, and the resulting filtrate was concentrated to give compound 13-7 (0.83 g, yellow liquid) in 82% yield. MS m/s (ESI): 278.2 [M+1].

[0305] In step 8: compound 11-4 (100 mg, 0.6 mmol) was dissolved in DCE (10 ml), compound 13-7 (169 mg, 0.6 mmol) and titanium isopropoxide (1 ml) were added and the reaction solution was stirred at 65°C for 12 h. Sodium borohydride (114 mg, 3 mmol) was added. The reaction solution was stirred at 65° C. for 6 hours. Water (20 ml) was added, the mixture was filtered, and the resulting filtrate was extracted with DCM (30 ml×2), then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting concentrate was purified by preparative chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give compound H-13 (30.01 mg, colorless liquid) in 11 .8%. MS m/s (ESI): 426.1 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 7,39 (дд, J=8,9, 5,3 Гц, 3H), 7,09-6.99 (м, 2H), 5,85 (дд, J=21,3, 1,5 Гц, 1H), 4,15-3,99 (м, 2H), 3,82-3,67 (м, 2H), 2,71-2,64 (м, 1H), 2,60-2,48 (м, 1H), 2,42-3,35 (м, 1H), 2,31 (д, J=14,0 Гц, 1H), 2,15 (qd, J=11.4, 4,9 Гц, 2H), 1,98-1,82 (м, 3H), 1,81-1,67 (м, 3H), 1,66-1.40 (м, 5H), 1,28-1,20 (м, 1H), 0,97 (т, J=12,9 Гц, 3H), 0,93-0,84 (м, 1H), 0,79-0,65 (м, 3H).

[0306] Example 14.

6-propyl-N-(2-(( R )-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-14)

[0307] In step 1: DMF (30 ml) was placed in a one-necked flask (100 ml), into which ethyl (E)-but-2-enoate (6 g, 42 mmol) was placed, potassium carbonate (8.7 g, 63 mmol) and pyrazole (4.3 g, 63 mmol) were added with stirring, and the reaction solution was stirred at 60°C for 36 h until the completion of the reaction, controlled using LC-MS. The reaction solution was filtered and the resulting filter cake was washed with EA (50 ml × 2), then washed with saturated brine (30 ml × 3), dried over anhydrous sodium sulfate, dried by centrifugation and purified by column chromatography (mobile phase - PE containing 5% EA) to obtain compound 14-1 (5.5 g, yellow liquid) with a yield of 6 2.3%. MS m/z (ESI): 211.1 [M+1].

[0308] In step 2: sodium hydroxide (1.52 g, 38 mmol) was dissolved in water (10 ml) and the resulting aqueous sodium hydroxide solution was pre-cooled to 5°C. Compound 14-1 (4 g, 19 mol), methanol (20 ml) and THF (20 ml) were placed in a flask (100 L), a pre-chilled aqueous solution of potassium hydroxide was added, the reaction solution was stirred for 2 h, concentrated hydrochloric acid was added to adjust the pH of the reaction solution to approximately 4, the reaction solution was extracted with DCM (100 ml × 2), all the resulting organic phases were combined and the combined organic phase was washed with brine, dried and concentrated to give 14-2 (3 g, white solid) in 86.7% yield. MS m/z (ESI): 183.1 [M+1].

[0309] In step 3: compound 14-2 (1.82 g, 10 mmol) and THF (40 ml) were placed in a three-necked flask (100 ml) under the protection of nitrogen gas and the mixture was cooled to -75°C. A 2.5 M solution of n-butyllithium in THF (10 mL, 25 mmol, 2.5 mol/L) was slowly added dropwise, and after completion of the dropwise addition, the reaction solution was stirred at -75°C for 3 h. the phase was washed with brine, dried and concentrated to give a brown liquid. The brown liquid was purified by column chromatography (mobile phase was PE containing 30% EA) to give compound 14-3 (100 mg, colorless liquid) in 6% yield. MS m/z (ESI): 165.1 [M+1].

[0310] In step 4: compound 14-3 (42 mg, 0.25 mmol) was dissolved in DCE (10 ml), compound 1a (65 mg, 0.25 mmol) and titanium isopropoxide (0.8 ml) were added, and the reaction solution was stirred at 58° C. for 12 h. Sodium borohydride (47.3 mg, 1.25 mmol) was added and the reaction solution was stirred at 45°C for 3 hours Was added water (20 ml), the mixture was filtered and the resulting filtrate was extracted with DCM (40 ml × 2), then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting concentrate was purified by preparative chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give compound H-14 (15.05 mg, white liquid) in 14.7 yield %. MS m/s (ESI): 409.2 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,55-8,49 (м, 1H), 7,81-7,73 (м, 1H), 7,54-7,48 (м, 1H), 7,39 (дд, J=5,8, 1,8 Гц, 1H), 7,27-7,20 (м, 1H), 5,83 (ддд, J=4,6, 1,9, 0,6 Гц, 1H), 4,16-4,00 (м, 2H), 3,80-3,72 (м, 2H), 2,95-2,78 (м, 1H), 2,67-2,48 (м, 2H), 2,47-2,37 (м, 1H), 2,16-1,97 (м, 3H), 1,89 (дд, J=13,2, 5,9 Гц, 1H), 1,85-1,64 (м, 4H), 1,64-1,30 (м, 8H), 1,13-1,04 (м, 1H), 0,99-0,88 (м, 3H), 0,79-0,71 (м, 1H).

[0311] Example 15.

6-(sec-butyl)-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-15)

[0312] In step 1: pyrazole (2.65 g, 39 mmol) was dissolved in a solution of DMF (60 ml), ethyl (E)-4-methylhexagonal-2-enoate (3 g, 19.5 mmol) and potassium carbonate (4.03 g, 29 mmol) and the reaction solution was stirred at 60°C overnight. EA (100 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:1) to give compound 15-1 (3 g) in 68% yield. MS m/s (ESI): 225.1 [M+1].

[0313] In step 2: compound 15-1 (2.6 g, 11.6 mmol) was dissolved in a mixture of methanol (20 ml) and water (4 ml), lithium hydroxide (0.8 g, 34.8 mmol) was added and the reaction solution was stirred at room temperature for 4 hours. hydrochloric acid solution to adjust the pH of the reaction solution to 2, the mixture was extracted with a mixture of DCM and methanol (10/1), and the resulting organic phase was distilled under reduced pressure to give a yellow solid. The yellow solid was purified by preparative liquid chromatography to give compound 15-2 (1.5 g) in 65% yield. MS m/h (ESI): 197.1 [M+1].

[0314] In step 3: Compound 15-2 (500 mg, 2.55 mmol) was dissolved in THF solution (20 mL), 2.5 M n-butyllithium solution (2.3 mL, 5.61 mmol) was added at -78°C, and the reaction solution was stirred at room temperature for 1 h. An aqueous solution of sodium chloride was added, the mixture was extracted with EA, the resulting organic phase was concentrated at reduced pressure, and the resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:1) to give compound 15-3 (180 mg) in 40% yield. MS m/s (ESI): 179.1 [M+1].

[0315] In Step 4: Compound 1a (116 mg, 0.45 mmol) was dissolved in a solution of DCE (5 ml), compound 15-3 (80 mg, 0.45 mmol) and titanium isopropoxide (1 ml) were added, and the reaction solution was heated to 60° C. to react overnight. After the reaction solution was cooled to room temperature, sodium borohydride (35 mg, 0.9 mmol) was added and the reaction solution was stirred at room temperature for 1 hour. Water was added, the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% change in acetonitrile) to give compound H-15 (34.96 mg) in 18% yield. MS m/s (ESI): 423.1 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6): δ 8,51-8,50 (д, J=4 Гц, 1H), 8,14 (с, 1H), 7,72-7,68 (м, 1H), 7,45-7,43 (м, 1H), 7,31-7,30 (м, 1H), 7,19-7,16 (м, 1H), 5,72-5,66 (м, 1H), 4,09-4,02 (м, 1H), 3,93-3,90 (м, 1H), 3,58-3,53 (м, 2H), 2,54-2,47 (м, 1H), 2,47-2,46 (м, 1H), 2,40-2,35 (м, 1H), 2,02-1,95 (м, 2H), 1,88-1,67 (м, 4H), 1,62-1,30 (м, 9H), 1,16-1,08 (м, 1H), 0,97-0,94 (м, 1H), 0,88-0,84 (т, J=8 Гц, 3H), 0,79-0,77 (т, J=4 Гц, 1H), 0,64-0,54 (м, 3H).

[0316] Example 16.

6-(1-methylcyclobutyl)-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-16)

[0317] In step 1: 1-methylcyclobutane-1-carboxylic acid (1.8 g, 12.7 mmol) was dissolved in a solution of DMF (20 ml), a solution of dimethylhydroxylamine hydrochloride (1.47 g, 15.2 mmol), HATU (7.2 g, 19 mmol) and triethylamine (3.85 g, 38.1 mmol) was added and the reaction solution was stirred at room temperature overnight. EA (100 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:1) to give compound 16-1 (2.1 g) in 100% yield. MS m/s (ESI): 158.1 [M+1].

[0318] In Step 2: Compound 16-1 (1.7 g, 10.8 mmol) was dissolved in THF solution (20 ml), lithium aluminum hydride (0.82 g, 21.6 mmol) was added at 0°C, and the reaction solution was stirred at 0°C for 1 h. Sodium sulfate decahydrate was added, the mixture was stirred for 30 min. and filtered, and the resulting filtrate was concentrated under reduced pressure to obtain compound 16-2.

[0319] In step 3: compound 16-2 (1.06 g, 40.8 mmol) was dissolved in a solution of acetonitrile (20 ml), ethyl diethoxyphosphinyl formate (3.63 g, 16.2 mmol), DBU (3.28 g, 21.6 mmol) and lithium chloride (0.91 g, 21.6 mmol) were added and the reaction solution was stirred at room temperature temperature during the night. EA (100 ml) was added, then washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=2:1) to give compound 16-3 (500 mg) in 28% yield. MS m/z (ESI): 169.1 [M+1].

[0320] In step 4: pyrazole (303 mg, 4.46 mmol) was dissolved in a DMF solution (10 ml), compound 16-3 (500 mg, 2.98 mmol) and potassium carbonate (822 mg, 5.96 mmol) were added and the reaction solution was stirred at 60°C overnight. EA (100 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=2:1) to give compound 16-4 (400 mg) in 57% yield. MS m/s (ESI): 237.1 [M+1].

[0321] In step 5: compound 16-4 (400 mg, 1.69 mmol) was dissolved in a mixture of methanol (20 ml) and water (2 ml), lithium hydroxide (78 mg, 3.39 mmol) was added and the reaction solution was stirred at room temperature for 2 hours. hydrochloric acid solution to adjust the pH of the reaction solution to 2, the mixture was extracted with a mixture of DCM and methanol (10/1), the resulting organic phase was distilled under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to obtain compound 16-5 (330 mg) with a yield of 94%. MS m/s (ESI): 209.1 [M+1].

[0322] In step 6: Compound 16-5 (270 g, 1.3 mmol) was dissolved in THF solution (10 ml), 2.5 M n-butyllithium solution (1.3 ml, 3.25 mmol) was added at -78° C., and the reaction solution was stirred at room temperature for 1 h. An aqueous solution of sodium chloride was added, the mixture was extracted with EA, the resulting organic phase was concentrated under reduced nom pressure and the resulting residue was purified by thin layer chromatography using eluents (PE: EA=1: 1) to obtain compound 16-6 (100 mg) with a yield of 40%. MS m/s (ESI): 191.1 [M+1].

[0323] In step 7: Compound 1a (136 mg, 0.53 mmol) was dissolved in a solution of DCE (10 ml), compound 16-6 (100 mg, 0.53 mmol) and titanium isopropoxide (0.5 ml) were added, and the reaction solution was reacted at 60° C. overnight. After the reaction solution was cooled to room temperature, sodium borohydride (40 mg, 1.06 mmol) was added and the reaction solution was stirred at room temperature for 1 hour. Water was added, the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% change in acetonitrile) to give compound H-16 (15.4 mg) in 6.7% yield. MS m/s (ESI): 435.1 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD): δ 8,50-8,49 (д, J=4 Гц, 1H), 7,69-7,67 (м, 1H), 7,44-7,42 (д, J=8 Гц, 1H), 7,29-7,27 (м, 1H), 7,18-7,15 (м, 1H), 5,71-5,65 (м, 1H), 4,14-4,12 (м, 1H), 3,86-3,83 (м, 1H), 3,62-3,57 (м, 2H), 2,65-2,63 (м, 1H), 2,47-2,45 (м, 2H), 2,30-2,29 (м, 2H), 2,05-2,03 (м, 1H), 1,97-1,89 (м, 2H), 1,84-1,73 (м, 4H), 1,63-1,53 (м, 6H), 1,41-1,30 (м, 5H), 0,92-0,89 (м, 4H).

[0324] Example 17.

N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-6-(1-(trifluoromethyl)cyclopropyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-17)

[0325] In step 1: 1-(trifluoromethyl)cyclopropane-1-carboxylic acid (6.8 g, 44 mmol) was dissolved in a solution of DMF (100 ml), a solution of dimethylhydroxylamine hydrochloride (5.14 g, 53 mmol), HATU (25 g, 66 mmol) and triethylamine (13.3 g, 132 mmol) was added, and the reaction solution was stirred at room temperature. temperature during the night. EA (400 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:1) to give compound 17-1 (8 g) in 92% yield. MS m/s (ESI): 198.1 [M+1].

[0326] In Step 2: Compound 17-1 (8 g, 4.06 mmol) was dissolved in THF solution (100 ml), DIBAL-H (1.0 M, 60 ml, 6.1 mmol) was added at -78°C and the reaction solution was stirred at -78°C for 1 h. Sodium sulfate decahydrate was added, the mixture was stirred for 30 min. and filtered, and the resulting filtrate was concentrated under reduced pressure to obtain compound 17-2.

[0327] In step 3: compound 17-2 (5.6 g, 40.6 mmol) was dissolved in a solution of acetonitrile (100 ml), ethyl diethoxyphosphinyl formate (13.64 g, 60.9 mmol), DBU (12.34 g, 81.2 mmol) and lithium chloride (3.4 g, 81.2 mmol) were added and the reaction solution was stirred at room temperature overnight. EA (300 ml) was added, then washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=2:1) to give compound 17-3 (4 g) in 47% yield. MS m/s (ESI): 209.1 [M+1].

[0328] In step 4: pyrazole (2.61 g, 38.5 mmol) was dissolved in a solution of DMF (100 ml), compound 17-3 (4 g, 19.2 mmol) and potassium carbonate (7.95 mg, 57.6 mmol) were added and the reaction solution was stirred at 60°C overnight. EA (300 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:1) to give compound 17-4 (3.3 g) in 62% yield. MS m/h (ESI): 277.1 [M+1].

[0329] In step 5: compound 17-4 (3.3 g, 12 mmol) was dissolved in a mixture of methanol (20 ml) and water (5 ml), lithium hydroxide (0.55 g, 24 mmol) was added and the reaction solution was stirred at room temperature for 2 hours. hydrochloric acid solution to adjust the pH of the reaction solution to 2, the mixture was extracted with a mixture of DCM and methanol (10/1), the resulting organic phase was distilled under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to obtain compound 17-5 (1.8 g) with a yield of 61%. MS m/s (ESI): 249.1 [M+1].

[0330] In step 6: compound 17-5 (1.8 g, 7.26 mmol) was dissolved in THF solution (30 ml), 2.5 M solution of n-butyllithium (7.3 ml, 18.1 mmol) was added at -78°C and the reaction solution was stirred at -20 for 1 h. centered under reduced pressure, and the resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:2) to give compound 17-6 (700 mg) in 42% yield. MS m/s (ESI): 231.1 [M+1].

[0331] In step 7: compound 17-6 (50 mg, 0.22 mmol), compound 1a (62 mg, 0.24 mmol) and titanium isopropoxide (1 ml) were dissolved in DCE (10 ml) and the reaction solution was reacted at 50° C. overnight. Sodium borohydride (60 mg, 1.6 mmol) was added and the reaction solution was reacted at 60° C. for 2 hours. Water (1 ml) was added to quench the reaction, the mixture was stirred at room temperature for 30 minutes, filtered, and the resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% changes in acetonitrile) to give compound H-17 (30.89 mg, colorless oily liquid) with 30% yield. MS m/s (ESI): 475.1 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD) δ 8,54-8,47 (м, 1H), 7,79-7,70 (м, 1H), 7,50 (дд, J=8,1, 1,0 Гц, 1H), 7,42 (дд, J=3,4, 1,9 Гц, 1H), 7,25-7,21 (м, 1H), 5,87 (ддд, J=16,6, 1,9, 0,7 Гц, 1H), 4,27 (т, J=6,7 Гц, 1H), 4,09-3,95 (м, 1H), 3,81-3,65 (м, 2H), 3,03-2,98 (м, 1H), 2,65-2,37 (м, 3H), 2,25-1,87 (м, 4H), 1,80-1,24 (м, 9H), 1,12-1,01 (м, 3H), 0,91-0,85 (м, 1H), 0,76-0,67 (м, 1H).

[0332] Example 18.

N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-6-(1,1,1,-trifluoro-2-methylprop-2-yl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-18)

[0333] In step 1: 3,3,3-trifluoro-2,2-dimethylpropanoic acid (5 g, 32 mmol) was dissolved in a solution of DMF (100 ml), a solution of dimethylhydroxylamine hydrochloride (3.74 g, 38.5 mmol), HATU (18.2 g, 48 mmol) and triethylamine (9.7 g, 96 mmol) was added and the reaction solution was stirred at room temperature overnight. EA (400 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:1) to give compound 18-1 (4.5 g) in 70% yield. MS m/s (ESI): 200.1 [M+1].

[0334] In Step 2: Compound 18-1 (4.3 g, 21.6 mmol) was dissolved in THF solution (30 ml), lithium aluminum hydride (1.64 g, 43.2 mmol) was added at 0°C and the reaction solution was stirred at 0°C for 1 h. Sodium sulfate decahydrate was added, the mixture was stirred for 30 min. and filtered, and the resulting filtrate was concentrated under reduced pressure to obtain compound 18-2.

[0335] In step 3: compound 18-2 (3.02 g, 21.6 mmol) was dissolved in a solution of acetonitrile (30 ml), ethyl diethoxyphosphinyl formate (7.26 g, 32.4 mmol), DBU (6.57 g, 43.2 mmol) and lithium chloride (1.81 g, 43.2 mmol) were added and the reaction solution was stirred at room temperature temperature during the night. EA (100 ml) was added, then washed with brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using eluents (PE: EA=2: 1) to give compound 18-3 (700 mg) in 15% yield. MS m/s (ESI): 211.1 [M+1].

[0336] In step 4: pyrazole (340 mg, 5 mmol) was dissolved in a solution of DMF (10 ml), the resulting compound 18-3 (700 mg, 3.33 mmol) and potassium carbonate (919 mg, 6.66 mmol) were added, and the reaction solution was stirred at 60°C overnight. EA (100 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (PE:EA=1:1) to give compound 18-4 (800 mg) in 86% yield. MS m/s (ESI): 279.1 [M+1].

[0337] In step 5: compound 18-4 (800 mg, 2.88 mmol) was dissolved in a mixture of methanol (10 ml) and water (2 ml), lithium hydroxide (0.13 g, 5.76 mmol) was added and the reaction solution was stirred at room temperature for 2 hours. hydrochloric acid solution to adjust the pH of the reaction solution to 2, the reaction mixture was extracted with a mixture of DCM and methanol (10/1), the resulting organic phase was distilled under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to obtain compound 18-5 (0.5 g) with a yield of 69%. MS m/s (ESI): 251.1 [M+1].

[0338] In step 6: Compound 18-5 (0.2 g, 0.8 mmol) was dissolved in THF solution (10 ml), 2.5 M n-butyllithium solution (0.8 ml, 2 mmol) was added at -78° C., and the reaction solution was stirred at room temperature for 1 h. Aqueous sodium chloride solution was added, the mixture was extracted with EA solution, the resulting organic phase was concentrated under reduced pressure and the resulting residue was purified by thin layer chromatography using eluents (PE: EA=1: 1) to obtain compound 18-6 (20 mg) in 10% yield. MS m/s (ESI): 233.1 [M+1].

[0339] In step 7: Compound 1a (23 mg, 0.09 mmol) was dissolved in a solution of DCE (5 ml), compound 18-6 (20 mg, 0.09 mmol) and titanium isopropoxide (0.3 ml) were added, and the reaction solution was heated to 60° C. to react overnight. After the reaction solution was cooled to room temperature, sodium borohydride (11 mg, 0.27 mmol) was added and the reaction solution was stirred at room temperature for 1 hour. Water was added, the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% change in acetonitrile) to give compound H-18 (1.65 mg) in 3.9% yield. MS m/h (ESI): 477.1 [M+1]; ¹ H ЯМР (400 МГц, CD ₃ OD): δ 8,53-8,52 (м, 1H), 7,77-7,73 (м, 1H), 7,52-7,50 (м, 1H), 7,42-7,40 (м, 1H),7,25-7,21 (м, 1H), 5,91-4,84 (м, 1H), 5,33-5,31 (т, J=4 Гц, 1H), 4,28-4,24 (т, J=4 Гц, 1H), 4,02-3,99 (м, 1H), 3,80-3,74 (м, 2H), 2,64-2,40 (м, 4H), 2,17-2,03 (м, 5H), 1,92-1,89 (д, J=12 Гц, 1H), 1,75-1,70 (м, 2H), 1,65 (с, 3H), 1,54-1,39 (м, 5H), 1,01-1,00 (д, J=4 Гц, 3H).

[0340] Example 19.

N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-4’,5’-dihydrospiro[cyclopentan-1,6’-pyrrolo[1,2-b]pyrazole]-4’-amine (H-19)

[0341] In step 1: A solution of acetonitrile (60 ml) and DBU (2.28 g, 15 mmol) was added to a sealed tube containing ethyl 2-cyclopentylidene acetate (1.54 g, 10 mmol) and pyrazole (1 g, 15 mmol). The mixture was stirred at 100°C for 18 hours, cooled to room temperature and concentrated, the resulting residue was diluted with ethyl acetate (100 ml), then washed with water (60 ml × 1), washed with brine (30 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=6/1) to give compound 19-1 (1.11 g, yellow oily liquid) in 50% yield. MS m/s (ESI): 223.1 [M+1].

[0342] In step 2: compound 19-1 (1.11 g, 5 mmol) was dissolved in a mixture of methanol (20 ml) and water (5 ml), sodium hydroxide (0.4 g, 10 mmol) was added, and the reaction solution was stirred at room temperature for 1 h until the reaction was completed. The reaction solution was concentrated, the resulting residue was dissolved in water (20 ml), 2 M hydrochloric acid was added to adjust the pH of the mixture from 3 to 4, the mixture was extracted with dichloromethane (40 ml × 3), the combined organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure to obtain compound 19-2 (0.77 g, yellow oily liquid) with the yield 80%. MS m/h (ESI): 195.2 [M+1].

[0343] In step 3: compound 19-2 (0.77 g, 4 mmol) was dissolved in dry tetrahydrofuran (15 ml), the mixture was cooled to -70°C under the protection of nitrogen gas, a 2.5 M solution of butyllithium in tetrahydrofuran (4 ml, 10 mmol) was added slowly dropwise. After completion of the dropwise addition, the reaction mixture was heated to -40° C. and stirred for 45 minutes. Water (10 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (petroleum ether/ethyl acetate=6/1) to give compound 19-3 (85 mg, yellow oily liquid) in 12% yield. MS m/h (ESI): 177.3 [M+1].

[0344] На шаге 4: соединение 19-3 (53 мг, 0,30 ммоль) и соединение 1а (78 мг, 0,30 ммоль) растворяли в 1,2-дихлорэтане (10 мл), добавляли изопропоксид титана (1 мл) и реакционный раствор перемешивали при 45°С в течение 18 ч. После охлаждения реакционного раствора до комнатной температуры добавляли боргидрид натрия (34 мг, 0,9 ммоль), реакционный раствор перемешивали в течение 3 ч, добавляли воду (5 мл), смесь перемешивали в течение 0,5 мин., фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной хроматографии (препаративная колонка: колонка С18 21,2 × 250 мм; система: 10 мМ NH ₄ HCO ₃ H ₂ O; длины волн: 254/214 нм; градиенты: 30%-60% изменений в ацетонитриле) с получением соединения H-19 (26 мг, бесцветное маслянистое вещество) с выходом 20,6%. MS m/s (ESI): 421.3 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,54-8,47 (м, 1H), 7,71-7,68 (м, 1H), 7,44 (д, J=8,0 Гц, 1H), 7,28 (дд, J=4,8, 1,8 Гц, 1H), 7,17 (дд, J=7,4, 4,8 Гц, 1H), 5,65 (дд, J=23,8, 1,7 Гц, 1H), 3,88 (дт, J=7,6, 5,3 Гц, 1H), 3,64-3,49 (м, 2H), 2,58-2,54 (м, 1H), 2,47-2,26 (м, 5H), 2,09-1,20 (м, 19H), 1,00-0,89 (м, 1H), 0,60-0,56 (м, 1H).

[0345] Example 20.

N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-2,3,4’,5,5’,6-hexahydrospiro[pyran-4,6’-pyrrolo[1,2-b]pyrazole]-4’-amine (H-20)

[0346] In step 1: A solution of acetonitrile (60 mL) and DBU (2.28 g, 15 mmol) was added to a sealed tube containing ethyl 2-(dihydro-2H-pyran-4(3H)-alkylene)acetate (1.7 g, 10 mmol) and pyrazole (1 g, 15 mmol). The mixture was stirred at 100°C for 18 hours, cooled to room temperature and concentrated, the resulting residue was diluted with ethyl acetate (100 ml), then washed with water (60 ml × 1), washed with brine (30 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=6/1) to give compound 20-1 (1.09 g, yellow oily liquid) in 46% yield. MS m/s (ESI): 239.1 [M+1].

[0347] In Step 2: Compound 20-1 (1.09 g, 40 mmol) was dissolved in a mixture of methanol (20 ml) and water (5 ml), sodium hydroxide (0.4 g, 10 mmol) was added, and the reaction solution was stirred at room temperature for 1 hour to complete the reaction. The reaction solution was concentrated, the resulting residue was dissolved in water (20 ml), 2 M hydrochloric acid was added to adjust the pH of the mixture from 3 to 4, the mixture was extracted with dichloromethane (40 ml × 3), the combined organic phase was dried over anhydrous sodium sulfate and filtered, the resulting filtrate was concentrated under reduced pressure to obtain compound 20-2 (0.73 g, yellow oily liquid) with the yield 76%. MS m/s (ESI): 211.2 [M+1].

[0348] In step 3: Compound 20-2 (0.73 g, 3.5 mmol) was dissolved in dry tetrahydrofuran (15 ml), and after cooling the mixture to -70°C under the protection of nitrogen gas, a 2.5 M solution of butyl lithium in tetrahydrofuran (3.5 ml, 8.7 mmol) was added slowly dropwise. After completion of the dropwise addition, the reaction solution was slowly warmed to -40° C. and stirred for 45 minutes. Water (10 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (petroleum ether/ethyl acetate=6/1) to give compound 20-3 (168 mg, yellow oily liquid) in 25% yield. MS m/h (ESI): 193.3 [M+1].

[0349] На шаге 4: соединение 20-3 (58 мг, 0,30 ммоль) и соединение 1а (78 мг, 0,30 ммоль) растворяли в 1,2-дихлорэтане (10 мл), добавляли изопропоксид титана (1 мл) и реакционный раствор перемешивали при 45°С в течение 18 ч. После охлаждения реакционного раствора до комнатной температуры добавляли боргидрид натрия (34 мг, 0,9 ммоль), реакционный раствор перемешивали в течение 3 ч, добавляли воду (5 мл), смесь перемешивали в течение 0,5 мин., фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной хроматографии (препаративная колонка: колонка С18 21,2 × 250 мм; система: 10 мМ NH ₄ HCO ₃ H ₂ O; длины волн: 254/214 нм; градиенты: 30%-60% изменений в ацетонитриле) с получением соединения H-20 (20 мг, бесцветное маслянистое вещество) с выходом 20,6%. MS m/h (ESI): 437.2 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,52-8,50 (м, 1H), 7,75-7,65 (м, 1H), 7,44 (дд, J=8,1, 1,2 Гц, 1H), 7,31 (дд, J=5,4, 1,8 Гц, 1H), 7,22-7,13 (м, 1H), 5,68 (дд, J=24,4, 1,7 Гц, 1H), 3,94-3,92 (м, 3H), 3,58-3,55 (м, 2H), 3,47-3,32 (м, 2H), 2,73-2,63 (м, 1H), 2,43-2,41 (м, 2H), 2,35-2,26 (м, 2H), 2,12-1,71 (м, 7H), 1,66-1,22 (м, 9H), 0,99-0,90 (м, 1H), 0,60-0,57 (м, 1H).

[0350] Example 21.

N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-4’,5’-dihydrospiro[cyclobutan-1,6’-pyrrolo[1,2-b]pyrazole]-4’-amine (H-21)

[0351] In step 1: cyclobutanone (3.5 g, 50 mmol) was dissolved in acetonitrile (100 ml), triethylphosphonoacetate (11.8 g, 52.5 mmol) and lithium chloride (2.2 g, 52.5 mmol) were added, the reaction solution was cooled from 2 to 5 ° C, DBU (7.5 g, 52.5 mmol) and the reaction solution were added slowly reacted at room temperature for 18 hours. The reaction solution was concentrated, the resulting residue was diluted with ethyl acetate (120 ml), then washed with water (70 ml × 1), washed with saturated saline (40 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1) to give compound 21-1 (5.6 g, yellow oily liquid) in 80% yield. MS m/h (ESI): 141.2 [M+1].

[0352] In step 2: a solution of acetonitrile (90 ml) and DBU (9.12 g, 60 mmol) was added to a sealed tube containing compound 21-1 (5.6 g, 40 mmol) and pyrazole (4 g, 60 mmol). The mixture was stirred at 100°C for 18 hours, cooled to room temperature and concentrated, the resulting residue was diluted with ethyl acetate (100 ml), then washed with water (60 ml × 1), washed with brine (30 ml × 1), dried over anhydrous sodium sulfate and filtered. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=6/1) to give compound 21-2 (5 g, yellow oily liquid) in 60% yield. MS m/s (ESI): 209.1 [M+1].

[0353] In step 3: compound 21-2 (5 g, 24 mmol) was dissolved in a mixture of methanol (60 ml) and water (15 ml), sodium hydroxide (1.92 g, 48 mmol) was added and the reaction solution was stirred at room temperature for 1 hour until the reaction was completed. The reaction solution was concentrated, the resulting residue was dissolved in water (40 ml), 2 M hydrochloric acid was added to adjust the pH of the mixture from 3 to 4, the mixture was extracted with dichloromethane (60 ml × 3), the combined organic phase was dried over anhydrous sodium sulfate and filtered, and the resulting filtrate was concentrated under reduced pressure to obtain compound 21-3 (3.0 g, yellow oily liquid) with the yield 70%. MS m/h (ESI): 181.2 [M+1].

[0354] In step 4: compound 21-3 (3.0 g, 16.7 mmol) was dissolved in dry tetrahydrofuran (80 ml), and after cooling the mixture to -70°C under the protection of nitrogen gas, a 2.5 M solution of butyllithium in tetrahydrofuran (16.7 ml, 41.7 mmol) was added slowly dropwise. After completion of the dropwise addition, the reaction solution was slowly warmed to -40° C. and stirred for 45 minutes. Water (10 ml) was added to quench the reaction, followed by filtration, concentration and purification by silica gel column chromatography (petroleum ether/ethyl acetate=6/1) to give compound 21-4 (243 mg, yellow oily liquid) in 9% yield. MS m/h (ESI): 163.3 [M+1].

[0355] На шаге 5: соединение 21-4 (49 мг, 0,30 ммоль) и соединение 1а (78 мг, 0,30 ммоль) растворяли в 1,2-дихлорэтане (10 мл), добавляли изопропоксид титана (1 мл) и реакционный раствор перемешивали при 45°С в течение 18 ч. После охлаждения реакционного раствора до комнатной температуры добавляли боргидрид натрия (34 мг, 0,9 ммоль), реакционный раствор перемешивали в течение 3 ч., добавляли воду (5 мл), смесь перемешивали в течение 0,5 ч., фильтровали, полученный фильтрат концентрировали при пониженном давлении и полученный остаток очищали с помощью препаративной хроматографии (препаративная колонка: колонка С18 21,2 × 250 мм; система: 10 мМ NH ₄ HCO ₃ H ₂ O; длины волн: 254/214 нм; градиенты: 30%-60% изменений в ацетонитриле) с получением соединения H-21 (8 мг, бесцветное маслянистое вещество) с выходом 6,6%. MS m/s (ESI): 407.3 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6) δ 8,51-8,49 (м, 1H), 7,74-7,64 (м, 1H), 7,43 (д, J=8,1 Гц, 1H), 7,34-7,31 (м, 1H), 7,18-7,15 (м, 1H), 5,66 (дд, J=17.7, 1,7 Гц, 1H), 3,86 (дт, J=7,6, 4,9 Гц, 1H), 3,64-3,49 (м, 2H), 2,79 (дд, J=13,0, 7,5 Гц, 1H), 2,64-2,50 (м, 1H), 2,46-2,24 (м, 5H), 2,17 (ддд, J=13,1, 5,1, 1,5 Гц, 1H), 2,06-2,01 (м, 2H), 1,97-1,65 (м, 5H), 1,65-1,18 (м, 8H), 0,99-0,88 (м, 1H), 0,59-0,55 (м, 1H).

[0356] Example 22.

6,6-dimethyl-N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-4-amine (H-22)

[0357] In step 1: 1H-pyrazole (2.68 g, 40 mmol) was dissolved in a solution of DMF (50 ml), ethyl 3-methyl-2-enoate (4.5 g, 40 mmol) and potassium carbonate (8.3 g, 60 mmol) were added and the reaction solution was stirred at 60°C overnight. Ethyl acetate (100 ml) was added, then washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (petroleum ether:ethyl acetate=1:1) to give compound 22-1 (1.2 g) in 15% yield. MS m/h (ESI): 197.1 [M+1].

[0358] In step 2: compound 22-1 (1.2 g, 6.2 mmol) was dissolved in a mixture of methanol (30 ml) and water (10 ml), lithium hydroxide (0.43 g, 18.6 mmol) was added and the reaction solution was stirred at room temperature for 4 h. hydrochloric acid solution to adjust the pH of the reaction solution to 2, the mixture was extracted with a mixture of dichloromethane and methanol (10/1), and the resulting organic phase was distilled under reduced pressure to obtain a yellow compound 22-2 (0.8 g). MS m/h (ESI): 169.1 [M+1].

[0359] In step 3: Compound 22-2 (0.8 g, 4.8 mmol) was dissolved in a solution of tetrahydrofuran (40 ml) and a 2.5 M solution of n-butyllithium (0.8 ml, 10.6 mmol) was added at -78°C, and the reaction solution was stirred at room temperature for 1 h. Aqueous sodium chloride solution was added, the mixture was extracted with ethyl acetate solution, the resulting organic the phase was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using eluents (petroleum ether:ethyl acetate=1:1) to give compound 22-3 (50 mg) in 7% yield. MS m/h (ESI): 151.1 [M+1].

[0360] In step 4: compound 1a (87 mg, 0.33 mmol) was dissolved in a solution of 1,2-dichloroethane (10 ml), compound 22-3 (50 mg, 0.33 mmol) and titanium isopropoxide (0.5 ml) were added, and the reaction solution was heated at 60° C. to react overnight. After the reaction solution was cooled to room temperature, sodium borohydride (26 mg, 0.66 mmol) was added and the reaction solution was stirred at room temperature for 1 hour. Water was added, the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography (preparative column: C18 column 21.2×250 mm; system: 10 mM NH ₄ HCO ₃ H ₂ O; wavelengths: 254/214 nm; gradients: 30%-60% change in acetonitrile) to give compound H-22 (2.27 mg) in 1.7% yield. MS m/s (ESI): 395.1 [M+1]; ¹ H ЯМР (400 МГц, ДМСО-d6): δ 8,51 (д, J=4Гц, 1H), 7,70-7,68 (м, 1H), 7,45-7,43 (д, J=8 Гц, 1H), 7,29-7,27 (м, 1H), 7,19-7,16 (м, 1H), 5,68-5,60 (м, 1H), 3,92-3,89 (м, 1H), 3,58-3,56 (м, 2H), 2,51-2,50 (м, 1H), 2,46-2,42 (м, 2H), 1,95-1,74 (м, 5H), 1,64-1,48 (м, 5H), 1,46-1,36 (м, 4H), 1,31-1,29 (м, 2H), 1,22-1,20 (м, 5H).

[0361] Example 23.

N-(2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decan-9-yl)ethyl)-4’,5’-dihydrospiro[cyclopropan-1,6’-pyrrolo[1,2-b]pyrazole]-4’-amine (H-23)

[0362] In step 1: 1H-pyrazole (6.8 g, 100 mmol) was dissolved in DMF (200 ml), tert-butyl-2-bromoacetate (21.8 g, 120 mmol) and potassium carbonate (20.7 g, 150 mmol) were added, and the reaction solution was stirred at room temperature for 4 hours. Ethyl acetate (400 ml) was added, then washed with saturated solution sodium bicarbonate, dried over anhydrous sodium sulfate and concentrated the organic phase under reduced pressure. The resulting residue was purified by thin layer chromatography using eluents (petroleum ether:ethyl acetate=3:2) to give compound 23-1 (7 g) in 38% yield. MS m/h (ESI): 183.1 [M+1].

[0363] На шаге 2: соединение 23-1 (7 г, 38,5 ммоль) растворяли в растворе тетрагирофурана (200 мл), добавляли раствор LiHMD (1 М, 85 мл) при -78°C, реакционный раствор перемешивали при -78°C в течение 1 ч., добавляли 1,3-диоксатиолан-2,2-диоксид (4,77 г, 38,5 ммоль) и реакционный раствор перемешивали при комнатной температуре в течение 3 ч. Добавляли раствор хлорида аммония, смесь экстрагировали раствором этилацетата, полученную органическую фазу концентрировали при пониженном давлении и полученный остаток очищали с помощью тонкослойной хроматографии с использованием элюентов (петролейный эфир:этилацетат=1: 1) с получением соединения 23-2 (5,4 г) с выходом 67,5%. MS m/s (ESI): 209.1 [M+1].

[0364] In step 3: Compound 23-2 (5.4 g, 26 mmol) was dissolved in tetrahydrofuran solution (100 ml), lithium aluminum hydride (2.96 g, 78 mmol) was added at 0°C, and the reaction solution was stirred at 0°C for 1 h. Sodium sulfate decahydrate was added, the reaction solution was stirred for 30 min. and filtered, the resulting filtrate was concentrated to give a solid. The solid was purified by thin layer chromatography using eluents (petroleum ether:ethyl acetate=1:1) to give compound 23-3 (2.2 g) in 61% yield. MS m/h (ESI): 139.1 [M+1].

[0365] In step 4: compound 23-3 (2.2 g, 16 mmol) was dissolved in dichloromethane solution (100 ml), triethylamine solution (3.23 g, 32 mmol) and methanesulfonyl chloride solution (2.76 g, 24 mmol) were added at 0°C and the reaction solution was stirred at 0°C for 1 h. Dichloromethane solution was added, then washed with 3 n. hydrochloric acid solution, washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give compound 23-4 (3 g, yellow solid). MS m/h (ESI): 217.1 [M+1].

[0366] In step 5: compound 23-4 (1.5 g, 6.9 mmol) was dissolved in a solution of acetonitrile (20 ml) and tetrabutylammonium cyanide (1.85 g, 6.9 mmol) and potassium carbonate (1.44 g, 10.4 mmol) were added. The reaction solution was heated under microwave conditions at 120° C. for 30 minutes. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using eluents (petroleum ether:ethyl acetate=1:1) to give compound 23-5 (800 mg) in 79% yield. MS m/s (ESI): 148.1 [M+1].

[0367] In step 6: compound 23-5 (800 mg, 5.4 mmol) was dissolved in ethanol solution (6 ml), potassium hydroxide solution (50%, 6 ml) was added and the reaction solution was stirred at 100° C. overnight. Added 3 n. hydrochloric acid solution to adjust the pH of the reaction solution to 2, the mixture was extracted with a mixture of dichloromethane and methanol (10/1), the resulting organic phase was distilled under reduced pressure to obtain a yellow solid (500 mg). The yellow solid was purified by preparative liquid chromatography to give compound 23-6 (150 g) in 18.5% yield. MS m/h (ESI): 167.1 [M+1].

[0368] In step 7: Compound 23-6 (150 mg, 0.9 mmol) was dissolved in a solution of tetrahydrofuran (10 ml) and a 2.5 M solution of n-butyllithium (0.8 ml, 10.6 mmol) was added at -78°C and the reaction solution was stirred at room temperature for 1 hour. Aqueous sodium chloride was added, the mixture was extracted with a solution of ethyl acetate, and the resulting the organic phase was concentrated under reduced pressure to give compound 23-7 . MS m/s (ESI): 149.1 [M+1].

[0369] In step 8: compound 1a (35 mg, 0.14 mmol) was dissolved in a solution of 1,2-dichloroethane (4 ml), compound 23-7 (20 mg, 0.14 mmol) and titanium isopropoxide (0.2 ml) were added, and the reaction solution was heated at 60° C. to react overnight. After the reaction solution was cooled to room temperature, sodium borohydride (11 mg, 0.28 mmol) was added and the reaction solution was stirred at room temperature for 1 hour. Water was added, the mixture was filtered, the resulting filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to give compound H-23 (0.45 mg) in 0.8% yield. MS m/h (ESI): 393.1 [M+1].

[0370] Bioanalysis

[0371] The cell lines used in the following assays are the PathHunter® CHO-K1 OPRM1 β-arrestin cell line from DiscoverX, Cat # 93-0213C2, Lot # 13K0402.

[0372] The reagents used, suppliers, catalog number, and storage temperature are listed below:

[0373] Assay Complete™ 107 Cell Culture Kit, DiscoverX, 92-3107G, -20°C;

[0374] Assay Complete™ Thaw Reagent, DiscoverX, 92-4002TR, -20°C;

[0375] Assay Complete™ Cell Separation Reagent, DiscoverX, 92-0009, -20°C;

[0376] Assay Complete™ Cell Coating Reagent, DiscoverX, 93-0563R2, -20°C;

[0377] Pathhunter Discovery Kit, DiscoverX, 93-0001, -20°C;

[0378] PBS (1×) 0.0067M (PO4), Hyclone, SH30256.01, 4°C;

[0379] DMSO, Sigma, D5879-100ML, room temperature;

[0380] NKH477, Sigma, 1603, -20°C; And

[0381] IBMX, Tocris, I5879, -20°C.

[0382] The tools, models, and vendors used are listed below:

[0383] Countsatr BioMed, IM1200, ALIT;

[0384] Microscope, IX51, OLYMPUS;

[0385] Centrifuge, 5804, Eppendorf;

[0386] Thermostatic water bath, DK-S420, Shanghai Shenxian thermostatic equipment factory;

[0387] Cell Incubator, 3111, Thermo;

[0388] Biological safety cabinet BSC-1300IIA2, AIRTECH;

[0389] White Opaque OptiPlate-384, 6007290, Perkin Elmer;

[0390] Victor X5 multimode tablet reader, PerkinElmer; And

[0391] White opaque culture plate-384 treated with TC, 6007680, PerkinElmer.

[0392] Test Example 1 HTRF-cAMP cell experiment

[0393] Experimental Methods and Steps

[0394] I. Cell resuscitation

[0395] 1. The resuscitation solution was removed from the refrigerator at 4°C and preheated over a water bath at 37°C for 15 minutes.

[0396] 2. The P6 cells were removed from the liquid nitrogen container, the frozen cell cryopreservation tube was immediately placed in a water bath at 37° C. and gently shaken for 30 seconds to 1 minute. until the appearance of small ice crystals or until the cells are completely thawed.

[0397] 3. The cryopreservation tube was carefully sterilized with 70% alcohol and wiped dry.

[0398] 4. Cells were centrifuged to remove cryoprotectant and resuspended in pre-warmed fresh resuscitation solution.

[0399] a. 3 ml of the pre-warmed cell resuscitation solution was aspirated and placed in a 15 ml centrifuge tube.

[0400] b. Cells were centrifuged at 1300 rpm. within 3 min.

[0401] c. The cryoprotectant supernatant was removed, and the cells were resuspended with 4 ml of pre-warmed resuscitation solution.

[0402] 5. The cell suspension was transferred to a T25 cell culture flask and cultured with 5% CO ₂ at 37°C for 24 hours.

[0403] 6. After culturing the cells for 24 hours, the resuscitation solution in the cell culture flask was replaced with pre-warmed cell culture medium.

[0404] II. Cell subculture

[0405] 1. After the cell growth density in the T25 culture flask exceeded 70%, the cells were digested with a cell dissociation solution for passage.

[0406] a. The culture flask medium was removed by aspiration, 4 ml of prewarmed PBS was added, the culture flask was gently shaken to wash the cells, and the PBS was removed by aspiration.

[0407] b. 1 ml of the cell dissociation solution was aspirated and added to a T25 culture flask.

[0408] c. The culture flask was shaken repeatedly to completely cover the culture flask with the dissociation solution and placed in a 5% CO ₂ incubator at 37° C. for 5 min.

[0409] The cell culture flask was removed and the cells were observed under a microscope to see if the cells had dissociated.

[0410] e. 3 ml of prewarmed cell culture medium was added to stop digestion.

[0411] e. The cell culture flask was gently washed again with the cell culture medium, the cell suspension was collected and placed in a 15 ml centrifuge tube.

[0412] f. Cells were centrifuged at 1300 rpm. within 3 min. and the supernatant was removed.

[0413] i. The cells were resuspended with 3 ml of cell culture medium.

[0414] 2. Cell subculture was performed at a ratio of 1:3 (i.e., 1 ml of cell resuspension solution and 3 ml of cell culture medium were placed in each T25 cell culture flask).

[0415] III. Seeding cells in a plate

[0416] 1. Step 2.2.1 (a to i) was repeated until P8 generation cells were obtained. Cells were counted and resuspended with 2×/1 mM IBMX stimulation buffer until the cell density was 1.2×10 ⁶ cells/ml.

[0417] 2. A solution of cells with a density of 1.2×10 ⁶ cells/ml was inoculated into a 384-well plate using a multichannel pipette at 10 μl/well (ie 12,000 cells/well).

[0418] IV. cAMP analysis

[0419] 1. Received the appropriate reagents, and the compounds were obtained in accordance with the tables for obtaining drug solutions.

[0420] a. 1× Stimulation Buffer: Add 1 ml of 5× Stimulation Stock Buffer and mix evenly with 4 ml of distilled water.

[0421] b. 2×/1 mM IBMX Stimulation Buffer (5 ml): 10 µl of 500 mM IBMX stock solution was added to 4990 µl of cell culture medium and mixed evenly by gentle pipetting.

[0422] c. Gradient dilution table for positive morphine drug:

Stock solution concentration / µM Volume from high concentration gradient (µl) Stimulation Buffer Volume (µl) Concentration after dilution / µM Volume of sample to be added to 384-well plate (µL) Final reaction concentration / μM 26606 - - - - - gradients 1 19 1330.3 - - 1) 2 104.4 25 8 10.0000 2) 20 40 8.3333 8 3.3333 3) 20 40 2.7778 8 1.1111 4) 20 40 0.9259 8 0.3704 5) 20 40 0.3086 8 0.1235 6) 20 40 0.1029 8 0.0412 7) 20 40 0.0343 8 0.0137 8) 20 40 0.0114 8 0.0046 9) 20 40 0.0038 8 0.0015 10) 20 40 0.0013 8 0.0005 eleven) 20 40 0.0004 8 0.0002

[0423] g. Prior to dilution, the compound was dissolved in DMSO to a storage concentration of 10 mM.

[0424] Dilution preparation table for TRV130 positive drug and various compounds:

Stock solution concentration / µM Volume from high concentration gradient (µl) Stimulation Buffer Volume (µl) Concentration after dilution / µM Volume of sample to be added to 384-well plate (µL) Final reaction concentration / μM 10000 - - - - - 1) 1 399 25 8 10.0000 2) 20 40 8.3333 8 3.3333 3) 20 40 2.7778 8 1.1111 4) 20 40 0.9259 8 0.3704 5) 20 40 0.3086 8 0.1235 6) 20 40 0.1029 8 0.0412 7) 20 40 0.0343 8 0.0137 8) 20 40 0.0114 8 0.0046 9) 20 40 0.0038 8 0.0015 10) 20 40 0.0013 8 0.0005 eleven) 20 40 0.0004 8 0.0002

[0425] e. 50 μM NK477 (1 ml): 1 μl of 50 mM NKH477 stock solution was added to 999 μl of 1x stimulation buffer and mixed uniformly by shaking.

[0426] Reagents for detection

[0427] A. cAMP-cryptate reaction solution (donor, lyophilized): 1 ml of 5x cAMP-cryptate stock solution was added to 4 ml of 1x lysis and detection buffer and gently mixed until homogeneous.

[0428] B. Anti-cAMP-d2 reaction solution (acceptor, lyophilized): 1 ml of 5x anti-cAMP-d2 stock solution was added to 4 ml of 1x lysis and detection buffer and gently mixed until homogeneous.

[0429] 2. Steps of cAMP analysis

[0430] a. 12,000 cells were inoculated into each well containing 10 μl of 2x IBMX Stimulation Buffer.

[0431] b. 8 µl of a diluted compound sample solution was added to the cells in each well.

[0432] c. 2 µl of prepared 10x NKH477 solution was added to each well.

[0433] d. Cells were incubated at 37°C for 45 minutes.

[0434] e. 10 µl of cAMP-d2 and 10 µl of anti-cAMP-cryptate reaction solution were added.

[0435] e. Cells were incubated in the dark at room temperature for 60 minutes.

[0436] f. The cell culture plate was read with HTRF.

[0437] 3. RFU detection and plate reading

[0438] After incubating the cells for 60 minutes. the plate with all samples was subjected to detection and read by homogeneous fluorescence with time resolution.

[0439] Data analysis

[0440] The data, which included two signal values at 665 nm and 620 nm, respectively, was exported from the appropriate software to a computer connected to a multi-mode plate reader. The formula for calculating the ratio was: ratio=signal value at 665 nm/signal value at 620 nm×10000. Data was analyzed using GraphPad Prism software. The best fitting curve was logarithm (agonist) vs. response. Compound EC50 values were determined by non-linear regression analysis of computerized dose-response curves; PEC50=-logEC50 (the unit of EC50 values was mol); % maximum morphine effect value = (compound sample-blank ratio)/TOP x 100 (Note: TOP is the top of the morphine sample-to-blank ratio curve, which was analyzed and fitted using Graphpad Prism software). The results are presented in table 1.

[0441] Table 1 Compound activity on cAMP

Compound cAMP
EC ₅₀ (nM) cAMP
Emax (%)
Morphine was 100% Compound cAMP
EC ₅₀ (nM) cAMP
Emax
Morphine was 100% H-1 49 124.6 H-2 47 93.2 H-3 17 74.6 H-4-1 8 64.1 H-4-2 18 91.8 H-6 9 99.6 H-7 29 104.6 H-10 eleven 74.7 H-11 15 84.8 H-12-1 12 99.5 H-12-2 12 77.8 H-14 31 73 H-15 84 64.9 H-17 31 68.6 H-18 68 89.1 H-19 21 94.1 H-20 4 107 H-21 22 70.8 H-22 eleven 104

[0442] Test Example 2 β -Arrestin Cell Experiment

[0443] Experimental method and steps

[0444] I. Cell resuscitation

[0445] 1. The resuscitation solution was removed from the refrigerator at 4°C and preheated in a water bath at 37°C for 15 minutes.

[0446] 2. The P6 cells were removed from the liquid nitrogen container, the frozen cell culture tube was immediately placed in a water bath at 37° C. and gently shaken for 30 seconds to 1 minute. until the appearance of small ice crystals or until the cells are completely thawed.

[0447] 3. The culture tube was carefully sterilized with 70% alcohol and wiped dry.

[0448] 4. Cells were centrifuged to remove cryoprotectant and resuspended in pre-warmed fresh resuscitation solution.

[0449] a. 3 ml of the pre-warmed cell resuscitation solution was aspirated and placed in a 15 ml centrifuge tube.

[0450] b. Cells were centrifuged at 1300 rpm. within 3 min.

[0451] c. The supernatant was removed and the cells were resuspended with 4 ml of pre-warmed resuscitation solution.

[0452] 5. The cell suspension was transferred to a T25 cell culture flask and cultured with 5% CO ₂ at 37°C for 24 hours.

[0453] 6. After culturing the cells for 24 hours, the resuscitation solution in the cell culture flask was replaced with pre-warmed cell culture medium.

[0454] II. Cell subculture

[0455] 1. After the cell growth density in the T25 culture flask exceeded 70%, the cells were digested with a cell dissociation solution for passage.

[0456] a. The culture flask medium was removed by aspiration, 4 ml of prewarmed PBS was added, the culture flask was gently shaken to wash the cells, and the PBS was removed by aspiration.

[0457] b. 1 ml of the cell dissociation solution was aspirated and added to a T25 culture flask.

[0458] c. The culture flask was shaken repeatedly to completely cover the culture flask with the dissociation solution and placed in a 5% CO ₂ incubator at 37° C. for 5 min.

[0459] The cell culture flask was removed and the cells were observed under a microscope to see if the cells had dissociated.

[0460] e. 3 ml of prewarmed cell culture medium was added to stop digestion.

[0461] e. The cell culture flask was gently washed again with the cell culture medium, the cell suspension was finally transferred to a 15 ml centrifuge tube.

[0462] f. Cells were centrifuged at 1300 rpm. within 3 min. and the supernatant was removed.

[0463] i. The cells were resuspended with 3 ml of cell culture medium.

[0464] 2. Cell subculture was performed at a ratio of 1:3 (i.e., 1 ml of cell resuspension solution and 3 ml of cell culture medium were placed in each T25 cell culture flask).

[0465] 3. Step 2.2.1 (a to i) was repeated until P8 generation cells were obtained.

[0466] III. Seeding cells in a plate

[0467] 1. 20 μl of the cell suspension was pipetted and the number of cells was measured using a cell counter.

[0468] 2. Cells were centrifuged at 1300 rpm. within 3 min. for cell sedimentation.

[0469] 3. The supernatant was removed and the appropriate cell culture solution was added until the cell density was 2×10 ⁵ cells/ml.

[0470] 4. Cell solution at a density of 2×10 ⁵ cells/ml was inoculated into a 384-well plate using a multichannel pipette at 20 μl/well (ie 4,000 cells/well).

[0471] 5. The cell-seeded 384-well plate was placed in a 5% CO ₂ incubator at 37°C for 24 hours.

[0472] IV. β-arrestin assay

[0473] 1. Compounds were prepared according to the following dilution tables.

[0474] a. Gradient dilution table for positive morphine drug:

Stock solution concentration / µM Volume from high concentration gradient (µl) Volume of culture buffer added (µL) Volume of DMSO added (µl) Concentration after dilution / µM Volume of sample to be added to 384-well plate (µL) Final reaction concentration / µM 26606 - - - - - - Preparation 2 18 0 2660.6 - - A 2 101.763 2.661 50,000 5 10,000 B 20 39 1 16.667 5 3.333 IN 20 39 1 5.556 5 1.111 G 20 39 1 1.852 5 0.370 D 20 39 1 0.617 5 0.123 E 20 39 1 0.206 5 0.041 AND 20 39 1 0.069 5 0.014 AND 20 39 1 0.023 5 0.005

[0475] b. Prior to dilution, the compound was dissolved in DMSO to a storage concentration of 10 mM.

[0476] Dilution preparation table for TRV130 positive drug and various compounds:

Stock solution concentration / µM Volume from high concentration gradient (µl) Volume of DMSO added (µl) Concentration after dilution / µM Volume of diluted solution (µl) Volume of culture buffer added (µL) Final concentration of the diluted solution / µM Volume of sample to be added to 384-well plate (µL) Final reaction concentration / μM 10000 - - - - - - - - A 12.5 50 2000 2.5 97.5 50,000 5 10,000 B 20 40 666.667 2.5 97.5 16.667 5 3.333 IN 20 40 222.222 2.5 97.5 5.556 5 1.111 G 20 40 74.074 2.5 97.5 1.852 5 0.370 D 20 40 24.691 2.5 97.5 0.617 5 0.123 E 20 40 8.230 2.5 97.5 0.206 5 0.041 AND 20 40 2.743 2.5 97.5 0.069 5 0.014 AND 20 40 0.914 2.5 97.5 0.023 5 0.005

[0477] 2. To a 384-well plate, 5 µl of diluted compound sample solution was added.

[0478] 3. After adding the samples, the 384-well plate was placed in a 5% CO ₂ incubator at 37° C. for 90 minutes.

[0479] V. RLU Definition

[0480] 1. Before completion of compound incubation, a detection working solution (in the dark) was prepared according to the following ratio. Then, 12.5 µl of detection working solution was added to each well, and the cells were incubated with shaking in the dark at room temperature for 1 hour.

[0481]

Ingredients Ratio Buffer for cell analysis 19 Substrate Reagent 1 5 Substrate Reagent 2 1 Overall volume 25

[0482] 2. After the end of compound incubation, 12.5 µl of the above working solution was added to each well, and the cells were incubated with shaking in the dark at room temperature and 80 rpm. within 1 hour

[0483] 3. After completion of the incubation, the plate was read using a multi-mode plate reader.

[0484] Data analysis

[0485] Data was exported from the appropriate software to a computer connected to a multi-mode plate reader and analyzed using GraphPad Prism software. The best fitting curve was logarithm (agonist) vs. response. Compound EC50 values were determined by non-linear regression analysis of computerized dose-response curves; PEC50=-logEC50 (the unit of EC50 values was mol); % maximum morphine effect value = (compound sample RLU value - empty well RLU value)/TOP×100 (Note: TOP is the top of the morphine sample RLU versus empty well RLU curve as analyzed and fitted using Graphpad Prism software). The results are presented in table 2.

[0486] Table 2. Results of testing compounds on β-arrestin

Compound β-arrestin
Emax (%)
Morphine was 100% Compound β-arrestin
Emax (%)
Morphine was 100% H-1 29.7 H-2 6.2 H-3 4.9 H-4-1 1.8 H-4-2 4.8 H-5 4.8 H-6 15.3 H-7 22.1 H-9 1.0 H-10 5.1 H-11 4.3 H-12-1 12 H-12-2 15.7 H-14 7.2 H-17 2.6 H-19 17.3 H-21 8.0 H-22 39.9

[0487] Table 1 and Table 2 show that representative compounds of the present disclosure have high cAMP inhibitory activity and high Emax values. In addition, the compounds of the present disclosure have small Emax values with respect to β-arrestin and have a good measurement error.

[0488] All documents referred to in this disclosure are incorporated herein by reference as if each document were separately incorporated by reference. In addition, it should be understood that after reading the above teaching content of this disclosure, various changes or modifications to this disclosure may be made by those skilled in the art, and these equivalent forms also fall within the scope of the appended formulas of this disclosure.

Claims (59)

1. The compound represented in formula (I), or a pharmaceutically acceptable salt, diastereomer, racemate, or mixture of diastereomers thereof: where ring A is pyridin-2-yl; (R ₀ ) _n means that the hydrogen atoms in ring A are replaced by n R ₀ groups, where n is 0; Y ₁ is N; Y ₂ is a bond; Y ₁ is directly connected to N; Y ₃ is CR ₃ ; Y ₄ is CR ₄ ; R ₃ and R ₄ are independently hydrogen or halogen; W is CH ₂ ; R _a , R _b , R _c and R _d are defined as follows: (i) each of R _a and R _b independently represents hydrogen or C _1-8 alkyl; or R _a and R _b together with their associated carbon atom form a three to six membered saturated monocyclic ring; R _c is hydrogen, halogen, C _1-8 alkyl, C _1-8 alkoxy, or halogenated C _1-8 alkyl; or R _d is hydrogen or C _1-8 alkyl; or (ii) each of R _a and R _b independently represents hydrogen; or R _c is linked to R _d to form a three to six membered saturated monocyclic ring or a four to six membered saturated monoheterocyclic ring, wherein the heteroatom in the four to six membered saturated monoheterocyclic ring is O or N; And m represents 0 or 1. 2. The compound or its pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers according to claim 1, where represents where the ring A, R ₀ and n are defined in Section 1. 3. A compound or a pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers thereof according to claim 1, wherein the compound represented in formula (I) has the structure represented by formula (II) where Z is N; (R ₀ ) _n means that the hydrogen atoms in the six-membered ring are substituted by n R ₀ groups, where n is 0; each of R _a and R _b is independently hydrogen or C _1-4 alkyl; or R _a and R _b together with their associated carbon atom form a three to six membered saturated monocyclic ring; R _c is hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, or halogenated C _1-4 alkyl; And Y ₁ , Y ₂ , Y ₃ , Y ₄ and m are defined in clause 1. 4. A compound or a pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers thereof according to claim 1, wherein Y ₃ is CH and Y ₄ is CH. 5. A compound or pharmaceutically acceptable salt, diastereomer, racemate, or mixture of diastereomers thereof according to claim 1, where m is 1. 6. A compound or a pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers thereof according to claim 3, wherein the compound represented in formula (II) has the structure represented by formula (III) 7. A compound or a pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers thereof according to claim 1, wherein R _a and R _b are each independently hydrogen or C _1-3 alkyl or R _a and R _b together with their associated carbon atom form a three to six membered saturated monocyclic ring. 8. A compound or pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers thereof according to claim 1, wherein R _c is hydrogen, halogen, C _1-3 alkyl, C _1-3 alkoxy, or halogenated C _1-3 alkyl. 9. A compound or a pharmaceutically acceptable salt, diastereomer, racemate or mixture of diastereomers thereof according to claim 1, wherein the compound is selected from the group consisting of: 10. An intermediate compound of a pyrrolopyrazole derivative substituted on the oxaspirocyclic ring, the intermediate compound having the structure represented by formula (I-2b) (I-2b), where R _a , R _b , R _c and R _d are defined as follows: (i) R _a and R _b are each independently hydrogen or C _1-8 alkyl; or R _a and R _b together with the carbon atom to which they are attached form a three to six membered saturated monocyclic ring; R _c is hydrogen, halogen, C _1-8 alkyl, C _1-8 alkoxy, or halogenated C _1-8 alkyl; A R _d is hydrogen or C _1-8 alkyl; or (ii) R _a and R _b are each independently hydrogen; And R _c is linked to R _d to form a three to six membered saturated monocyclic ring or a four to six membered saturated monoheterocyclic ring, wherein the heteroatom in the four to six membered saturated monoheterocyclic ring is O or N; And m is 0 or 1; wherein when m is 0 and one of R _c and R _d is hydrogen, the other R _c and R _d is not hydrogen. 11. An intermediate compound according to claim 10, wherein the compound represented by formula (I-2b) has the structure represented by formula (I-2a) where R _a , R _b and R _c are as defined in clause 10. 12. An intermediate compound according to claim 10, where each of R _a and R _b independently represents hydrogen or C _1-3 alkyl; or R _a and R _b together with their associated carbon atom form a three to six membered saturated monocyclic ring. 13. Intermediate according to claim 10, wherein R _c is hydrogen, halogen, C _1-3 alkyl, C _1-3 alkoxy, or halogenated C _1-3 alkyl. 14. An intermediate compound according to claim 10, wherein the compound represented in formula (I-2b) is selected from the group consisting of the following structures: