WO2021027304A1

WO2021027304A1 - Analgesic compound, preparation method therefor, and pharmaceutical use thereof

Info

Publication number: WO2021027304A1
Application number: PCT/CN2020/084773
Authority: WO
Inventors: 刘力锋; 胡斌; 谢婧; 石晓永; 赵金柱; 何宛; 郭淑春
Original assignee: 上海海雁医药科技有限公司; 扬子江药业集团有限公司
Priority date: 2019-08-14
Filing date: 2020-04-14
Publication date: 2021-02-18
Also published as: CN112789276A

Abstract

The present invention relates to a bi-heterocyclic methylethylamine substituted oxaspiro derivative, a preparation method therefor, and a pharmaceutical use thereof. In particular, disclosed are a compound of formula (I) or a pharmaceutically acceptable salt, a stereoisomer or a solvate thereof, a preparation method therefor and the use thereof. The definition of each group in the formula is detailed in the description and claims.

Description

Analgesic compound, its preparation method and medical use

Technical field

The present invention relates to a class of biheterocyclic methylethylamine substituted oxaspiro derivatives, a preparation method thereof, a pharmaceutical composition containing the derivatives, and a therapeutic agent, especially as a MOR receptor agonist and in preparation Use in medicines for the treatment and prevention of pain and pain related diseases.

Background technique

Opioid receptors are an important type of G protein-coupled receptor (GPCR), which is the target for the binding of endogenous opioid peptides and opioid drugs. Endogenous opioid peptides are naturally produced in mammals. Opioid active substances, currently known endogenous opioid peptides are roughly divided into enkephalins, endorphins, dynorphins and neoorphins. There are corresponding opioid receptors in the central nervous system, namely μ (MOR), δ (DOR), κ (KOR) receptors and so on. Studies have found that the analgesic effect of endogenous opioid peptides is mainly determined by the expression of opioid receptors, which are the targets of opioid drugs and endogenous opioid peptides.

Current research suggests that GPCR mediates and regulates physiological functions through two main pathways: the G protein pathway and the β-arrestin pathway. After the traditional GPCR agonist binds to the receptor, it activates the G protein signal pathway, including calcium ion and other second messenger systems, adenyl cyclase (AC), and mitogen-activated protein kinase (mitogen-activated protein). kinases, MAPK), etc., and β-arrestin preferential ligands mainly activate the β-arrestin pathway. The β-arrestin-mediated GPCR reaction mainly includes three aspects: 1) As a negative regulator, it interacts with G protein-coupled receptor kinase (GRK) to desensitize GPCRs and stop G protein signal transduction. 2) As a scaffold protein, it recruits endocytosis proteins and induces GPCR endocytosis; 3) As a linker protein, it forms a complex with downstream signal molecules of GPCRs and activates signal transduction molecules in a G protein-independent manner, Such as MAPK, Src protein tyrosine kinase and Akt. The difference of ligand-stimulated G protein signal and/or β-arrestin signal ultimately determines the ligand-specific cellular biological effects of GPCR.

MOR is the target of opioid analgesics such as endogenous enkephalin and morphine. Early studies have shown that endogenous enkephalins and the opioid drug etorphine can stimulate G protein and trigger receptor endocytosis, but morphine does not trigger receptor endocytosis at all. This is because morphine stimulates MOR phosphorylation. The ability is too weak and can only recruit a small amount of β-arrestin on the membrane (Zhang et al., Proc Natl Acad Sci USA, 1998, 95(12): 7157-7162). Such ligands perform their physiological functions entirely through the G protein signaling pathway instead of the β-arrestin pathway. Studies have found that after injection of morphine into β-arrestin2 knockout mice, the analgesic effect mediated by G protein signal is stronger and lasts longer (Bohn et al., Science, 1999). It is foreseeable that if the negative β-arrestin preference of such ligands is stronger, and it can even escape β-arrestin-mediated receptor desensitization, it may lead to prolonged G protein signal transmission time and produce a more powerful analgesic effect. .

Currently published patent applications for MOR agonists include WO2017106547, WO2017063509, WO2012129495, WO2017106306, and so on.

Long-term use of opioids can produce side effects such as tolerance, respiratory depression and constipation, and these side effects have been shown to be closely related to the function of β-arrestin. In order to reduce the side effects of opioids, drugs can be designed based on the negative β-arrestin preference ligand of MOR to reduce the side effects mediated by β-arrestin and enhance the therapeutic effect.

Summary of the invention

The purpose of the present invention is to provide a compound with a novel structure that can be used as a MOR receptor agonist.

The first aspect of the present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt, stereoisomer or solvate thereof:

Where

R ₀ is hydrogen or substituted or unsubstituted C _1-10 alkyl (preferably substituted or unsubstituted C _1-6 alkyl, more preferably substituted or unsubstituted C _1-3 alkyl);

R ₁ and R ₂ are each independently hydrogen, halogen, substituted or unsubstituted C _1-10 alkyl (preferably substituted or unsubstituted C _1-6 alkyl, more preferably substituted or unsubstituted C _{1- 3} alkyl);

Or R ₁ , R ₂ and the connected carbon atoms together form a 3 to 6 membered saturated or unsaturated monocyclic ring or a 3 to 6 membered saturated or unsaturated monocyclic ring; the 3 to 6 membered saturated or unsaturated monocyclic ring, The 3- to 6-membered saturated or unsaturated monocyclic ring is unsubstituted or substituted with 1-3 substituents selected from the group consisting of halogen, C _1-10 alkoxy (preferably C _1-6 alkoxy, more Preferably C _1-3 alkoxy), C _1-10 alkyl (preferably C _1-6 alkyl, more preferably C _1-3 alkyl), halogenated C _1-10 alkyl (preferably halogen (C _1-6 alkyl, more preferably halogenated C _1-3 alkyl);

R ₃ and R ₄ are each independently hydrogen, hydroxy, cyano, halogen, substituted or unsubstituted C _1-10 alkyl (preferably substituted or unsubstituted C _1-6 alkyl, more preferably substituted or unsubstituted Substituted C _1-3 alkyl), substituted or unsubstituted C _1-10 alkoxy (preferably substituted or unsubstituted C _1-6 alkoxy, more preferably substituted or unsubstituted C _1-3 Alkoxy), halogenated C _1-10 alkyl (preferably halogenated C _1-6 alkyl, more preferably halogenated C _1-3 alkyl), substituted or unsubstituted C _3-8 cycloalkyl (Preferably substituted or unsubstituted C _3-6 cycloalkyl), NR ₁₁ R ₁₂ , or substituted or unsubstituted 4 to 6-membered saturated monocyclic heterocyclic ring;

Or R ₃ , R ₄ and the connected carbon atoms together form a 3 to 6 membered saturated or unsaturated monocyclic ring or a 3 to 6 membered saturated or unsaturated monocyclic ring; the 3 to 6 membered saturated or unsaturated monocyclic ring, The 3- to 6-membered saturated or unsaturated monocyclic ring is unsubstituted or substituted with 1-3 substituents selected from the group consisting of halogen, C _1-10 alkoxy (preferably C _1-6 alkoxy, more Preferably C _1-3 alkoxy), C _1-10 alkyl (preferably C _1-6 alkyl, more preferably C _1-3 alkyl), halogenated C _1-10 alkyl (preferably halogen (C _1-6 alkyl, more preferably halogenated C _1-3 alkyl);

R ₁₁ and R ₁₂ are each independently hydrogen, substituted or unsubstituted C _1-10 alkyl (preferably substituted or unsubstituted C _1-6 alkyl, more preferably substituted or unsubstituted C _1-3 alkane Group), halogenated C _1-10 alkyl (preferably halogenated C _1-6 alkyl, more preferably halogenated C _1-3 alkyl), substituted or unsubstituted C _3-8 cycloalkyl (preferably Is a substituted or unsubstituted C _3-6 cycloalkyl), or a substituted or unsubstituted 3 to 6-membered saturated or unsaturated monocyclic heterocyclic ring; or R ₁₁ , R ₁₂ and the connected nitrogen atom form a substituted or unsubstituted 4- to 6-membered saturated or unsaturated monocyclic heterocyclic ring;

X is O or NR _c ; R _c is hydrogen or C _1-10 alkyl (preferably C _1-6 alkyl, more preferably C _1-3 alkyl);

R _a , R _b and the connected carbon atoms together form a substituted or unsubstituted C _6-10 aromatic ring, or a substituted or unsubstituted 5 or 6-membered monocyclic heteroaromatic ring, the C _6-10 aromatic ring or 5 Or a 6-membered monocyclic heteroaromatic ring and the connected heterocyclic ring form a fused bicyclic ring;

The "substitution" means that 1, 2, or 3 hydrogen atoms in the group are replaced by substituents independently selected from Group A; the substituents of Group A are selected from: cyano, acetyl, hydroxyl, Hydroxymethyl, hydroxyethyl, carboxyl, halogenated C _1-8 alkyl (preferably halogenated C _1-6 alkyl, more preferably halogenated C _1-3 alkyl), halogen (preferably F or Cl ), nitro, C _6-10 aryl (preferably phenyl), 5- or 6-membered monocyclic heteroaryl, C _1-10 alkyl (preferably C _1-6 alkyl, more preferably C _1-3 Alkyl), C _1-10 alkoxy (preferably C _1-6 alkoxy, more preferably C _1-3 alkoxy), C _3-8 cycloalkyl (preferably C _3-6 cycloalkane Group), C _3-8 cycloalkoxy (preferably C _3-6 cycloalkoxy), C _2-10 alkenyl (preferably C _2-6 alkenyl, more preferably C _2-4 alkenyl) , C _2-10 alkynyl (preferably C _2-6 alkynyl, more preferably C _2-4 alkynyl), NR _a0 R _b0 , -CONR _a0 R _b0 , -C(O)OC _1-10 alkyl (Preferably -C(O)OC _1-6 alkyl, more preferably -C(O)OC _1-3 alkyl), -CHO, -OC(O)C _1-10 alkyl (preferably -OC (O)C _1-6 alkyl, more preferably -OC(O)C _1-3 alkyl), -SO ₂ C _1-10 alkyl (preferably -SO ₂ C _1-6 alkyl, more preferably Is -SO ₂ C _1-3 alkyl), -SO ₂ C _6-10 aryl (preferably -SO ₂ C ₆ aryl, such as -SO ₂ -phenyl), -COC _6-10 aryl (preferably Is -COC ₆ aryl, such as -CO-phenyl), 4 to 6-membered saturated or unsaturated monocyclic ring or 4 to 6-membered saturated or unsaturated monocyclic ring, wherein the C _6-10 aryl, 5 or 6-membered monocyclic heteroaryl, 4 to 6-membered saturated or unsaturated monocyclic ring or 4 to 6-membered saturated or unsaturated monocyclic ring is unsubstituted or is selected from acetyl, hydroxyl, cyanide by 1, 2 or 3 Group, halogen, C _1-3 alkyl, C _1-3 alkoxy, C _3-6 cycloalkyl, NR _a0 R _b0 substituent substituted; R _a0 , R _b0 are each independently hydrogen or C _{1- 3} alkyl.

In another preferred embodiment, the group A substituent is selected from: cyano, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, halogenated C _1-3 alkyl, halogen (preferably F or Cl ), nitro, phenyl, 5- or 6-membered monocyclic heteroaryl, C _1-3 alkyl, C _1-3 alkoxy, C _3-6 cycloalkyl, C _3-6 cycloalkoxy, C _2-4 alkenyl, C _2-4 alkynyl, NR _a0 R _b0 , -CONR _a0 R _b0 , -C(O)OC _1-3 alkyl, -CHO, -OC(O)C _1-3 alkane Group, -SO ₂ C _1-3 alkyl, -SO ₂ -phenyl, -CO-phenyl, 4 to 6 membered saturated or unsaturated monocyclic ring or 4 to 6 membered saturated or unsaturated monocyclic ring, wherein R _a0 and R _b0 are each independently hydrogen or C _1-3 alkyl.

In another preferred embodiment, R ₀ is hydrogen or C _1-3 alkyl (preferably methyl).

In another preferred embodiment, R ₀ is hydrogen.

In another preferred example, R ₁ and R ₂ are each independently hydrogen or C _1-3 alkyl (preferably methyl).

In another preferred example, R ₁ and R ₂ are each independently hydrogen.

In another preferred embodiment, the 4- to 6-membered saturated or unsaturated monocyclic heterocyclic ring in the substituent group A is selected from: azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole , Piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, 1,2-dihydroazetidine, 1,2- Dihydrooxetadiene, 2,5-dihydro-1H-pyrrole, 2,5-dihydrofuran, 2,3-dihydrofuran, 2,3-dihydro-1H-pyrrole, 3,4 -Dihydro-2H-pyran, 1,2,3,4-tetrahydropyridine, 3,6-dihydro-2H-pyran or 1,2,3,6-tetrahydropyridine.

In another preferred example, the 4- to 6-membered saturated or unsaturated monocyclic ring in the substituent group A is selected from: cyclobutyl ring, cyclopentyl ring, cyclopentenyl ring, cyclohexyl ring, cyclohexenyl Ring, cyclohexadienyl ring.

In another preferred embodiment, the 5- or 6-membered monocyclic heteroaryl group in the group A substituent is selected from: thiophene, N-alkyl pyrrole, furan, thiazole, imidazole, oxazole, pyrrole, pyrazole, three Azole, 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.

In another preferred example, the 3- to 6-membered saturated or unsaturated monocyclic heterocyclic ring formed by R ₁ , R ₂ and the connected carbon atoms is selected from: aziridine, ethylene oxide, azetidine, oxa Cyclobutane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, 1,2- Dihydroazetadiene, 1,2-dihydrooxetadiene, 2,5-dihydro-1H-pyrrole, 2,5-dihydrofuran, 2,3-dihydrofuran, 2 ,3-Dihydro-1H-pyrrole, 3,4-dihydro-2H-pyran, 1,2,3,4-tetrahydropyridine, 3,6-dihydro-2H-pyran, 1,2, 3,6-Tetrahydropyridine.

In another preferred example, the 3- to 6-membered saturated or unsaturated monocyclic ring formed by R ₁ , R ₂ and the connected carbon atoms is selected from: cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, and cyclopentenyl ring , Cyclohexyl ring, cyclohexenyl ring, cyclohexadienyl ring.

In another preferred example, the 3- to 6-membered saturated or unsaturated monocyclic heterocyclic ring formed by R ₃ , R ₄ and the connected carbon atoms is selected from: aziridine, ethylene oxide, azetidine, oxa Cyclobutane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, 1,2- Dihydroazetadiene, 1,2-dihydrooxetadiene, 2,5-dihydro-1H-pyrrole, 2,5-dihydrofuran, 2,3-dihydrofuran, 2 ,3-Dihydro-1H-pyrrole, 3,4-dihydro-2H-pyran, 1,2,3,4-tetrahydropyridine, 3,6-dihydro-2H-pyran, 1,2, 3,6-Tetrahydropyridine.

In another preferred embodiment, the 3- to 6-membered saturated or unsaturated monocyclic ring formed by R ₃ , R ₄ and the connected carbon atoms is selected from: cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, and cyclopentenyl ring , Cyclohexyl ring, cyclohexenyl ring, cyclohexadienyl ring.

In another preferred embodiment, the 4- to 6-membered saturated monocyclic heterocyclic ring in R ₃ and R ₄ is selected from: azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine Pyridine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran.

In another preferred embodiment, the 4- to 6-membered saturated or unsaturated monocyclic heterocyclic ring formed by R ₁₁ , R ₁₂ and the connected nitrogen atom is selected from: azetidine, tetrahydropyrrole, piperidine, piperazine, Morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, 1,2-dihydroazetidine, 2,5-dihydro-1H-pyrrole, 2,3-dihydro -1H-pyrrole, 1,2,3,4-tetrahydropyridine, 1,2,3,6-tetrahydropyridine.

In another preferred embodiment, the 3- to 6-membered saturated or unsaturated monocyclic heterocyclic ring in R ₁₁ and R ₁₂ is selected from: aziridine, ethylene oxide, azetidine, and oxetane , Tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, 1,2-dihydronitrogen Cyclobutadiene, 1,2-dihydrooxetadiene, 2,5-dihydro-1H-pyrrole, 2,5-dihydrofuran, 2,3-dihydrofuran, 2,3- Dihydro-1H-pyrrole, 3,4-dihydro-2H-pyran, 1,2,3,4-tetrahydropyridine, 3,6-dihydro-2H-pyran, 1,2,3,6 -Tetrahydropyridine.

In another preferred example, the 5- or 6-membered monocyclic heteroaryl group described in the substituent group A is selected from the following structures:

The above-mentioned 5- to 6-membered monocyclic heteroaryl group is optionally substituted with 1, 2 or 3 substituents each independently selected from Group A.

In another preferred embodiment, the C _6-10 aromatic ring formed by _Ra , R _b and the connected carbon atoms is a benzene ring.

In another preferred embodiment, the 5- or 6-membered monocyclic heteroaromatic ring formed by _Ra , _Rb and the connected carbon atoms is thiophene or furan.

In another preferred example, the C _3-8 cycloalkyl group in R ₁₁ and R ₁₂ is selected from the group consisting of cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, and cyclohexyl ring.

In another preferred example, the C _3-8 cycloalkyl group in R ₃ and R ₄ is selected from the group consisting of cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, and cyclohexyl ring.

In another preferred example, the 5- or 6-membered monocyclic heteroaromatic ring formed by _Ra , _Rb and the connected carbon atoms is selected from the following structures:

among them

The two ring atoms connected are represented by adjacent pairs of atoms shared when fused with other rings. The 5- or 6-membered monocyclic heteroaromatic ring is unsubstituted or substituted with 1, 2, or 3 substituents each independently selected from Group A.

In another preferred embodiment, R ₃ and R ₄ are each independently hydrogen, C _1-3 alkyl, C _1-3 alkyl substituted with C _1-3 alkoxy, C _3-6 cycloalkyl, NH _2. NH(C _1-3 alkyl) or N(C _1-3 alkyl) ₂ ;

Or R ₃ , R ₄ and the connected carbon atoms together form a 4- to 6-membered saturated monocyclic ring or a 3- to 6-membered saturated monocyclic ring; the 4- to 6-membered saturated monocyclic ring or a 3- to 6-membered saturated monocyclic ring is not Substituted or substituted with 1-3 substituents selected from the group consisting of halogen, C _1-3 alkoxy, C _1-3 alkyl, halo C _1-3 alkyl.

In another preferred example, R ₃ and R ₄ are each independently hydrogen, methyl, ethyl, n-propyl, isopropyl, N(CH ₃ ) ₂ , methoxyethyl, cyclopropyl, and cyclopropyl. Butyl or cyclopentyl; or R ₃ , R ₄ and the connected carbon atoms together form a tetrahydropyran ring, a cyclopropyl ring, a cyclobutyl ring, a cyclopentyl ring or a cyclohexyl ring.

In another preferred example, X is O or NR _c ; R _c is hydrogen or methyl.

In another preferred example, X is O.

In another preferred example,

Selected from the following structures:

X, R ₀ , R ₃ , and R ₄ are as defined in the specification.

In another preferred example,

Selected from the following structures:

In another preferred example,

Selected from the following structures:

In another preferred embodiment, the 3- to 6-membered or 4- to 6-membered saturated monocyclic heterocyclic ring is selected from the following structures:

The hydrogen atoms on the above-mentioned 3- to 6-membered or 4- to 6-membered saturated monocyclic heterocyclic ring are optionally substituted with 1, 2 or 3 substituents each independently selected from Group A.

In another preferred embodiment, the compound is selected from Table A, wherein the compound of Table A is selected from the following group:

In another preferred embodiment, the compound is selected from:

The second aspect of the present invention provides a pharmaceutical composition comprising the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof; and a pharmaceutically acceptable a.

The third aspect of the present invention provides the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the pharmaceutical composition according to the second aspect of the present invention in the preparation of preventive and / Or use in drugs for treating related diseases mediated by MOR receptor agonists.

The fourth aspect of the present invention provides the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, or the pharmaceutical composition according to the second aspect of the present invention is prepared for Use in drugs that agonize or antagonize MOR receptors.

The fifth aspect of the present invention provides the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the pharmaceutical composition according to the second aspect of the present invention in the preparation of prevention and / Or use in medicines for the treatment of pain and pain-related diseases.

In another preferred embodiment, the related diseases mediated by the MOR receptor agonist are selected from pain, immune dysfunction, inflammation, esophageal reflux, neurological and mental diseases, urinary and reproductive diseases, cardiovascular diseases and respiratory diseases, preferably pain.

In another preferred example, the pain is selected from postoperative pain, pain caused by cancer, neuropathic pain, traumatic pain, and pain caused by inflammation.

In another preferred embodiment, the cancer is selected from breast cancer, endometrial cancer, cervical cancer, skin cancer, prostate cancer, ovarian cancer, fallopian tube tumor, ovarian tumor, hemophilia and leukemia.

The sixth aspect of the present invention provides a method for preventing and/or treating related diseases mediated by MOR receptor agonists, comprising administering to a patient a therapeutically effective amount of the compound according to the first aspect of the present invention or its pharmacy Above acceptable salt, stereoisomer or solvate, or pharmaceutical composition as described in the second aspect of the invention.

The seventh aspect of the present invention provides a method for preventing and/or treating pain and pain-related diseases, comprising administering to a patient a therapeutically effective amount of the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt thereof , Stereoisomers or solvates, or pharmaceutical compositions as described in the second aspect of the present invention.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as the embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, I will not repeat them here.

detailed description

After extensive and in-depth research, the inventors unexpectedly discovered that this type of bi-heterocyclic methyl ethylamine-substituted oxaspiro derivatives not only have excellent analgesic effects, but also have good preference. The compound of the invention has excellent pharmacokinetic properties. Therefore, the series of compounds are expected to be developed as drugs for the treatment and prevention of pain and pain-related diseases. On this basis, the inventor completed the present invention.

Definition of Terms

As used herein, "alkyl" refers to linear and branched saturated aliphatic hydrocarbon groups, C _1-10 alkyl is an alkyl group containing 1 to 10 carbon atoms, preferably C _1-6 alkyl, more preferably It is a C _1-3 alkyl group with similar definitions; non-limiting examples of alkyl groups include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl , N-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3 -Methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl Butyl, 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, 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 , And various branched chain isomers are more preferred.

As used herein, "cycloalkyl" and "cycloalkyl ring" are used interchangeably, and both refer to a saturated or partially unsaturated monocyclic cyclic hydrocarbon group, and "C _3-8 cycloalkyl" refers to containing 3 to 8 The carbon atom cyclic hydrocarbon group is preferably a C _3-6 cycloalkyl group, and the definition is similar. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl , Cyclooctyl, etc., preferably cyclopropyl, cyclopentyl, and cyclohexenyl.

As used herein, "C _1-10 alkoxy" refers to -O-(C _1-10 alkyl), where the definition of alkyl is as described above. C _1-6 alkoxy is preferable, and C _1-3 alkoxy is more preferable. Non-limiting examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, isobutoxy, pentoxy and the like.

As used herein, "C _3-8 cycloalkoxy" refers to -O-(C _3-8 cycloalkyl), wherein cycloalkyl is defined as described above. Preferred is C _3-6 cycloalkoxy. Non-limiting examples include cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy and the like.

As used herein, "C _6-10 aryl" and "C _6-10 aryl ring" are used interchangeably, and both refer to all-carbon monocyclic or fused polycyclic rings with a conjugated π-electron system (that is, sharing adjacent The ring) group of a carbon atom pair refers to an aryl group containing 6 to 10 carbon atoms; phenyl and naphthyl are preferred, and phenyl is more preferred.

As used herein, "a bond" means that two groups connected by it are connected by a covalent bond.

As used herein, "halogen" refers to fluorine, chlorine, bromine or iodine.

As used herein, "halo" refers to the replacement of one or more (eg, 1, 2, 3, 4, or 5) hydrogens in a group with halogen.

For example, "halo C _1-10 alkyl" means that an alkyl group is substituted with one or more (such as 1, 2, 3, 4, or 5) halogens, where the definition of alkyl is as described above. It is selected as a halogenated C _1-6 alkyl group, more preferably a halogenated C _1-3 alkyl group. Examples of halogenated C _1-8 alkyl include (but are not limited to) monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, Monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, etc.

For another example, "halogenated C _1-10 alkoxy" means that the alkoxy group is substituted with one or more (such as 1, 2, 3, 4, or 5) halogens, wherein the definition of alkoxy is as described above. It is preferably a halogenated C _1-6 alkoxy group, and more preferably a halogenated C _1-3 alkoxy group. Including (but not limited to) trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethoxy and the like.

For another example, "halo C _3-8 cycloalkyl" refers to a cycloalkyl group substituted with one or more (such as 1, 2, 3, 4, or 5) halogens, wherein the definition of cycloalkyl is as described above. Preferably, it is a halogenated C _3-6 cycloalkyl group. Including (but not limited to) trifluorocyclopropyl, monofluorocyclopropyl, monofluorocyclohexyl, difluorocyclopropyl, difluorocyclohexyl and the like.

As used herein, "amino" refers to NH _2, "cyano" refers to the CN, "Nitro" refers to NO _2, "benzyl" refers to -CH ₂ - phenyl, "oxo" refers to = O, "carboxy" Refers to -C(O)OH, "acetyl" refers to -C(O)CH ₃ , "hydroxymethyl" refers to -CH ₂ OH, and "hydroxyethyl" refers to -CH ₂ CH ₂ OH or -CHOHCH ₃ , "Hydroxy" refers to -OH, "thiol" refers to SH, and the structure of "cyclopropylene" is:

As used herein, "heteroaryl ring" and "heteroaryl" are used interchangeably and refer to having 5 to 10 ring atoms, preferably 5 or 6 membered monocyclic heteroaryl or 8 to 10 membered bicyclic heteroaryl ; The ring array shares 6, 10 or 14 π electrons; and in addition to carbon atoms, there are groups with 1 to 5 heteroatoms. "Heteroatom" refers to nitrogen, oxygen, or sulfur.

As used herein, "3 to 6 membered (4 to 6 membered) saturated or partially unsaturated monocyclic ring" refers to a saturated or partially unsaturated all-carbon monocyclic ring containing 3 to 6 ring atoms. Examples of 3 to 6-membered saturated or partially unsaturated monocyclic rings include (but are not limited to): cyclopropyl ring, cyclobutyl ring, cyclopentyl ring, cyclopentenyl ring, cyclohexyl ring, cyclohexenyl ring, ring Hexadienyl ring and so on.

As used herein, "3 to 6 membered (4 to 6 membered) saturated or partially unsaturated monocyclic heterocyclic ring" means that 1, 2 or 3 carbon atoms in the 3 to 6 membered monocyclic ring are selected from nitrogen, oxygen or S (O) _t (where t is an integer of 0 to 2) substituted by heteroatoms, but not including the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon; preferably 4 to 6 members, more It is preferably 5 to 6 yuan. Examples of 3- to 6-membered saturated or partially unsaturated monocyclic heterocycles include (but are not limited to) propylene oxide, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, pyrrole Morpholine, oxazolidine, piperazine, dioxolane, dioxane, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, 1,2-di Hydroazetidine, 1,2-dihydrooxetadiene, 2,5-dihydro-1H-pyrrole, 2,5-dihydrofuran, 2,3-dihydrofuran, 2, 3-Dihydro-1H-pyrrole, 3,4-dihydro-2H-pyran, 1,2,3,4-tetrahydropyridine, 3,6-dihydro-2H-pyran, 1,2,3 ,6-Tetrahydropyridine and so on.

As used herein, "5- to 6-membered monocyclic heteroaryl ring" and "5- to 6-membered monocyclic heteroaryl" are used interchangeably, and both refer to a mono-heteroaryl ring containing 5 to 6 ring atoms, Examples include (but are not limited to): thiophene ring, N-alkane pyrrole ring, furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole 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-oxadiazole ring, 1,3,4-oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine Ring, pyrimidine ring, pyrazine ring, etc.

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

Unless otherwise defined, the "substituents independently selected from ..." in the present invention means that when more than one hydrogen on a group is substituted by a substituent, the types of the substituents may be the same or different, so The selected substituents are of independent types.

Unless otherwise defined, the "...same or different, and each independently is..." in the present invention means that when there are more than one identical substituent groups in the general formula, the groups may be the same or different, each Independent species. For example, L is (CR ₀₁ R ₀₂ ) _s , when s is 2, that is, L is (CR ₀₁ R ₀₂ )-(CR ₀₁ R ₀₂ ), and the two R ₀₁ or R ₀₂ can be the same or different. Independent type, for example, L can 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 ).

As used herein, any group herein may be substituted or unsubstituted. When the above groups are substituted, the substituents are preferably 1 to 5 or less groups independently selected from CN, halogen, C _1-10 alkyl (preferably C _1-6 alkyl, more preferably C _1-3 Alkyl), C _1-10 alkoxy (preferably C _1-6 alkoxy, more preferably C _1-3 alkoxy), halogenated C _1-8 alkyl (preferably halogenated C _{1- 6} alkyl, more preferably halogenated C _1-3 alkyl), C _3-8 cycloalkyl (preferably C _3-6 cycloalkyl), halogenated C _1-8 alkoxy (preferably halogenated C _1-6 alkoxy, more preferably halogenated C _1-3 alkoxy), C _1-8 alkyl substituted amino, amino, halogenated C _1-8 alkyl substituted amino, acetyl Group, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, nitro, C _6-10 aryl (preferably phenyl), C _3-8 cycloalkoxy (preferably C _3-6 cycloalkoxy), C _2-10 alkenyl (preferably C _2-6 alkenyl, more preferably C _2-4 alkenyl), C _2-10 alkynyl (preferably C _2-6 alkynyl, more preferably C _2-4 Alkynyl), -CONR _a0 R _b0 , -C(O)OC _1-10 alkyl (preferably -C(O)OC _1-6 alkyl, more preferably -C(O)OC _1-3 alkyl ), -CHO, -OC(O)C _1-10 alkyl (preferably -OC(O)C _1-6 alkyl, more preferably -OC(O)C _1-3 alkyl), -SO ₂ C _1-10 alkyl (preferably -SO ₂ C _1-6 alkyl, more preferably -SO ₂ C _1-3 alkyl), -SO ₂ C _6-10 aryl (preferably -SO ₂ C ₆ Aryl, such as -SO ₂ -phenyl), -COC _6-10 aryl (preferably -COC ₆ aryl, such as -CO-phenyl), 4 to 6-membered saturated or unsaturated monocyclic heterocyclic ring, 4 to 6-membered saturated or unsaturated monocyclic ring, 5- to 6-membered monocyclic heteroaryl ring, 8- to 10-membered bicyclic heteroaryl ring, spiro ring, spiro heterocyclic ring, bridged ring or bridged heterocyclic ring, wherein R _a0 , R _b0 Each is independently hydrogen or C _1-3 alkyl. .

The various substituent groups described herein above can themselves be substituted by the groups described herein.

When the 4- to 6-membered (5- to 6-membered) saturated monocyclic heterocyclic ring described herein is substituted, the positions of the substituents may be in their possible chemical positions. Representative substitution situations of exemplary monocyclic heterocyclic rings are shown below :

Wherein "Sub" represents the various substituents described herein;

Represents the connection with other atoms.

Unless otherwise defined, when the 4- to 6-membered saturated monocyclic heterocyclic ring in the present invention is a substituent, it may itself be substituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, Hydroxy, C _1-3 alkyl, O=, NR _a0 R _b0 , hydroxymethyl, hydroxyethyl, carboxyl, -C(O)OC _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, two Oxolane, 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, pyrazine Ring; wherein R _a0 and R _b0 are each independently hydrogen or C _1-3 alkyl.

The "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic acid or an organic acid that can retain the biological effectiveness of the free base without other side effects.

"Pharmaceutically acceptable base addition salts" include, but are not limited to, salts of inorganic bases such as sodium, potassium, calcium and magnesium salts. Including but not limited to salts of organic bases, such as ammonium salt, triethylamine salt, lysine salt, arginine salt and the like.

The "solvate" mentioned in the present invention refers to a complex formed by the compound of the present invention and a solvent. They either react in a solvent or precipitate or crystallize out of the solvent. For example, a complex formed with water is called a "hydrate". Solvates of compounds of formula (I) fall within the scope of the present invention.

The compound represented by formula (I) or formula (II) of the present invention may contain one or more chiral centers and exist in different optically active forms. When a compound contains a chiral center, the compound contains enantiomers. The present invention includes these two isomers and mixtures of isomers, such as racemic mixtures. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When the compound of formula (I) or formula (II) contains more than one chiral center, diastereomers may exist. The present invention includes the resolved optically pure specific isomers and mixtures of diastereomers. Diastereoisomers can be resolved by methods known in the art, such as crystallization and preparative chromatography.

The present invention includes prodrugs of the aforementioned compounds. Prodrugs include known amino protecting groups and carboxyl protecting groups, which are hydrolyzed under physiological conditions or released through enzymatic reactions to obtain the parent compound. For specific preparation methods of prodrugs, please refer to (Saulnier, MG; Frennesson, DB; Deshpande, MS; Hansel, SB and Vysa, DMBioorg. Med. Chem Lett. 1994, 4, 1985-1990; and Greenwald, RB; Choe, YH; Conover, CD; Shum, K.; Wu, D.; Royzen, MJ Med. Chem. 2000, 43, 475.).

Generally, the compound of the present invention or its pharmaceutically acceptable salt, or its solvate, or its stereoisomer, or prodrug can be administered in a suitable dosage form with one or more pharmaceutically acceptable carriers. These dosage forms are suitable for oral, rectal, topical, intraoral, and other parenteral administration (for example, subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules, and syrups. The compounds of the present invention contained in these formulations may be solid powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; water-in-oil or oil-in-water emulsions, and the like. The above-mentioned dosage forms can be prepared from the active compound and one or more carriers or excipients through general pharmaceutical methods. The above-mentioned carrier needs to be compatible with the active compound or other excipients. For solid preparations, commonly used non-toxic carriers include but are not limited to mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose and the like. Carriers for liquid preparations include water, physiological saline, aqueous dextrose, ethylene glycol, polyethylene glycol, and the like. The active compound can form a solution or a suspension with the aforementioned carriers.

The composition of the present invention is formulated, quantified and administered in a manner that conforms to medical practice standards. The "therapeutically effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual to be treated, the cause of the condition, the target of the drug, and the mode of administration.

As used herein, "therapeutically effective amount" refers to the amount of the compound of the present invention that will cause an individual's biological or medical response, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease, etc. the amount.

The therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a stereoisomer thereof contained in the pharmaceutical composition of the present invention is preferably 0.1 mg-5 g/kg (body weight).

As used herein, "pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid substance or liquid filling machine, diluent, encapsulating material or auxiliary preparation or any type of excipient, which is compatible with the patient and most It is preferably a mammal, more preferably a human, which is suitable for delivering the active agent to the target target without terminating the activity of the agent.

As used herein, "patient" refers to an animal, preferably a mammal, and more preferably a human. The term "mammal" refers to warm-blooded spinal mammals, including cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, and humans.

As used herein, "treating" refers to reducing, delaying progression, attenuating, preventing, or maintaining an existing disease or condition (e.g., cancer). Treatment also includes curing one or more symptoms of the disease or condition, preventing its development, or alleviating to a certain degree.

Preparation

The experimental methods without specific conditions in the following examples usually follow the conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions described by the manufacturer The suggested conditions.

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

The compound represented by the following formula (I) can be prepared by a known method, for example, by the following method, a method equivalent thereto, or the method described in the examples. In the following preparation method, the raw material compound may be in the form of a salt, and the salt may be any pharmaceutically acceptable salt exemplified by the compound represented by formula (I) of the present invention.

(In each formula of the above scheme, all groups are defined as described in the specification.)

Specifically, the compound represented by formula (I) can be prepared according to the following method: (R)-2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl) Acetaldehyde and the compound represented by the formula (Ia) undergo a reductive amination reaction to prepare the compound represented by the formula (I).

The reductive amination reaction conditions are known and can be, for example, in an organic solvent (such as DCM, DCE or THF, etc.), under the catalysis of a catalyst (such as tetraisopropyl titanate), using a reducing agent (such as boron) Sodium hydride), the carbonyl group undergoes reductive amination reaction with amine.

The compound having an amino group, a carboxyl group, or a hydroxyl group used in the present invention can be prepared by using a compound that has been protected by a protective group commonly used for this group as required. After passing through the reaction process of the above-mentioned reaction scheme, a known desorption can be carried out. Protection response.

Compared with the prior art, the main advantages of the present invention are:

A series of oxaspiro derivatives substituted with biheterocyclic methylethylamine with novel structures are provided, which have higher inhibitory activity on cAMP, higher Emax value, and excellent analgesic effect. In addition, the present invention The compound has a lower Emax value for β-arrestin, and has a good bias. Therefore, it can be developed into drugs for the treatment and prevention of pain and pain-related diseases.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods that do not indicate specific conditions in the following examples usually follow the conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight. Unless otherwise defined, the terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to the content described can be applied to the present invention.

As used herein, DMB is 2,4-dimethoxybenzyl, THF is tetrahydrofuran, EA is ethyl acetate, PE is petroleum ether, Ac ₂ O is acetic anhydride, NBS is N-bromosuccinimide, DCM is dichloromethane, AIBN is azobisisobutyronitrile, Pd(dppf)Cl ₂ is 1,1'-bis(diphenylphosphorferrocene]palladium dichloride, TFA is trifluoroacetic acid, TBSCl Is tert-butyldimethylchlorosilane, NCS is N-chlorosuccinimide, DHP is dihydrotetrahydropyran, LiAlH ₄ is lithium aluminum hydride, PMB is p-methoxybenzyl, LiHMDS is two (Trimethylsilyl) lithium amide, Pd ₂ (dba) ₃ is tris(dibenzylideneacetone) dipalladium, RuPhos is 2-dicyclohexylphosphorus-2',6'-diisopropoxy-1 ,1'-biphenyl, DMAP is 4-dimethylaminopyridine, THP is tetrahydrotetrahydropyran, n-BuLi is n-butyllithium, TMsOTf is trimethylsilyl trifluoromethanesulfonate, TEBAC is tri Ethyl benzyl ammonium chloride, HATU is 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, DMF is dimethylformamide , DMSO is dimethyl sulfoxide, DIEA is N,N-diisopropylethylamine, and BINAP is (2R,3S)-2,2'-bisdiphenylphosphino-1,1'-binaphthyl.

As used herein, room temperature refers to about 20-25°C.

Example 1: N-((5'H,7'H-spiro[cyclopentane-1,4'-thieno[2,3-c]pyran]-7'-yl)methyl)- 2-((R)-9-(pyridin-2-yl)-6-oxospiro[4.5]dec-9-yl)ethylamine (H-1)

Step 1: Dissolve methyl 2-(thiophen-3-yl)acetate (300mg, 1.92mmol) in 10mL of anhydrous tetrahydrofuran, cool to -70℃ with a dry ice-ethanol bath, and add hexamethyldisilylamine dropwise Lithium (1M tetrahydrofuran solution, 2.1mL, 2.1mmol), after the addition is complete, continue stirring for 1 hour, add 1,4-dibromobutane (412mg, 1.92mmol) dropwise, warm to 0°C and continue stirring for 2 hours, Recool to -70°C, add dropwise lithium hexamethyldisilazide (1M tetrahydrofuran solution, 2.1mL, 2.1mmol), warm to 0°C and continue stirring for 2 hours, then use saturated aqueous ammonium chloride solution (50mL) for the reaction Quench, extract twice with ethyl acetate (50mL X 2). The organic phases were combined, washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude methyl 1-(thiophen-3-yl)cyclopentane-1-carboxylate (400 mg, yellow oil) , Used directly in the next reaction Yield: 99%. MS m/z(ESI): 211.1[M+1].

Step 2: Dissolve methyl 1-(thiophen-3-yl)cyclopentanecarboxylate (400mg) in 10mL of anhydrous tetrahydrofuran, cool to 0℃ in an ice water bath, and add tetrahydrolithium aluminum (217mg, 5.71mmol) ). The reaction solution was stirred overnight at room temperature and then recooled to 0°C, quenched with ethyl acetate (5mL) and sodium sulfate decahydrate (400mg), stirred for 10 minutes, filtered, and concentrated under reduced pressure to obtain crude product (1-(thiophen-3) -Cyclopentyl) methanol (300 mg, yellow oil), used directly in the next reaction. Yield: 86%. MS m/z(ESI): 183.1[M+1].

Step 3: Combine (1-(thiophen-3-yl)cyclopentyl)methanol (300mg, 1.65mmol), 2,2-diethoxyethylamine (438mg, 3.30mmol), trifluoromethanesulfonic acid (2mL A mixture of) and 1,4-dioxane (10 mL) was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was poured into a cold saturated aqueous sodium bicarbonate solution, extracted twice with ethyl acetate (50mL x 2), combined the organic phases, washed with brine (50mL), dried with anhydrous sodium sulfate, and filtered. Concentrated under reduced pressure to obtain crude product (5'H,7'H-spirocyclo[cyclopentane-1,4'-thieno[2,3-c]pyran]-7'-yl)methylamine (200mg, Yellow oil), directly used in the next reaction. Yield: 54%. MS m/z(ESI): 224.1[M+1].

Step 4: Combine (5',7'-dihydrospirocyclopentane-1,4'-thieno[2,3-c]pyran]-7'-yl)methylamine (50mg) and (R )-2-(9-(pyridin-2-yl)-6-oxospiro[4.5]dec-9-yl)acetaldehyde (58mg, 0.22mmol), sodium cyanoborohydride (42 mg, 0.67mmol) The mixture of methanol and methanol (5mL) was stirred overnight at room temperature, concentrated under reduced pressure, and separated and purified by liquid phase preparation to obtain the target product N-((5'H,7'H-spirocyclo[cyclopentane-1,4' -Thieno[2,3-c]pyran]-7'-yl)methyl)-2-((R)-9-(pyridin-2-yl)-6-oxospiro[4.5]dec- 9-yl)ethylamine H-1 (19.5 mg, white solid), yield: 19%. MS m/z(ESI): 467.2[M+1]. 1H NMR(400MHz,DMSO-d6)δ8.49(s,1H), 8.20(s,1H), 7.67-7.69(m,1H),7.41-7.43(m,1H),7.25-7.27(m,1H) ),7.16(s,1H),6.88-6.90(m,1H),4.57(s,1H),3.57-3.62(m,4H),3.26-3.36(m,2H),2.47-2.68(m,3H) ),2.28-2.40(m,2H),1.73-1.77(m,2H),1.30-1.1.62(m,15H),0.59-0.92(m,2H)

Example 2: N-((5'H,7'H-spiro[cyclohexane-1,4'-thieno[2,3-c]pyran]-7'-yl)methyl)-2 -((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine (H-2)

Step 1: Dissolve methyl 2-(thiophen-3-yl)acetate (1.56g, 0.01mol) in 40mL N,N-dimethylformamide, add 1,5-diiodopentane (4.86g, 0.015mol), cooling to 2-5°C, adding NaH (60%, 1.2g, 0.03mol) in batches, and stirring at room temperature for 1 hour. 80mL of water was added to the reaction solution, extracted with ethyl acetate (30mL×3), the organic phases were combined, washed with saturated brine (40mL×1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and used a silica gel column The residue obtained was purified by chromatography with eluent (petroleum ether/ethyl acetate=4/1) to obtain methyl 1-(thiophen-3-yl)cyclohexanecarboxylate (1.2 g, yellow oily liquid) as Rate: 53.6%. MS m/z(ESI): 225.3[M+1].

Step 2: To a suspension of lithium tetrahydroaluminum (0.41 g, 10.7 mmol) in methyl tert-butyl ether (35ml) was slowly added methyl 1-(thiophen-3-yl)cyclohexanecarboxylate (1.2g ) In methyl tert-butyl ether (10ml), the mixture was stirred at room temperature for 1.5 hours, 10mL ice water was added to the reaction solution, stirred vigorously for 5 minutes, filtered, separated into layers, the organic phase was washed with saturated brine (15mL× 1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent (petroleum ether/ethyl acetate = 2/1) to obtain (1-(thiophen-3) -Cyclohexyl) methanol (0.76 g, yellow oily liquid), yield: 72.4%. MS m/z(ESI): 178.3[M-18].

Step 3: Add dioxane (30ml) to (1-(thiophen-3-yl)cyclohexyl)methanol (1g, 3.88mmol) and 2,2-dimethoxyethylamine (0.48g, 4.26mmol) Trifluoromethanesulfonic acid (2ml) solution was slowly added to the solution, the mixture was stirred at room temperature for 1.5 hours, 40mL ice water was added to the reaction solution, pH was adjusted to 9-10 with saturated potassium carbonate, and ethyl acetate extraction (30ml* 3), the combined organic phase was washed with saturated brine (15mL×1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the eluent (petroleum ether/ethyl acetate=2/ 1) Purify the resulting residue to obtain the product (5'H,7'H-spiro[cyclohexane-1,4'-thieno[2,3-c]pyran]-7'-yl)methylamine (0.65g, yellow oily liquid), yield: 71%. MS m/z(ESI): 238.3[M+1].

Step 4: Add N-((5'H,7'H-spiro[cyclohexane-1,4'-thieno[2,3-c]pyran]-7'-yl)methylamine (60mg , 0.25mmol) and (R)-2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)acetaldehyde (66mg, 0.25mmol) dissolved in 8mL1,2 -To dichloroethane, add 0.5 mL of tetraisopropyl titanate and stir for 18 hours at 45°C. Cool to room temperature, add sodium borohydride (34 mg, 0.9 mmol) to the reaction solution, stir for 3 hours, and add to the reaction solution 5mL of water was added, stirred for 0.5 minutes, filtered, the filtrate was concentrated under reduced pressure, and the residue obtained was purified by preparative chromatography to obtain the product N-((5'H,7'H-spiro[cyclohexane-1,4'-thieno[ 2,3-c]pyran]-7'-yl)methyl)-2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl ) Ethylamine H-2 (20mg, brown solid), yield: 17%. MS m/z (ESI): 481.3 [M+1]. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.49 (dd ,J=5.0,1.8Hz,1H),7.69-7.65(m,1H),7.42(d,J=8.1Hz,1H),7.25-7.11(m,2H),6.98(d,J=5.2Hz, 1H),4.50-4.45(m,1H),4.09(d,J=11.4Hz,1H),3.59-3.56(m,2H),3.25-3.22(m,2H),2.63-2.59(m,1H) , 2.47-2.22 (m, 5H), 1.95-1.11 (m, 20H), 0.95-0.92 (m, 1H), 0.59-0.54 (m, 1H).

Example 3: N-((5'H,7'H-spiro[cyclobutane-1,4'-thieno[2,3-c]pyran]-7'-yl)methyl)-2 -((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine (H-3)

The preparation method is the same as in Example 1, except that the 1,4-dibromobutane in step 1 is replaced with 1,3-dibromopropane, MS m/z(ESI): 453.2[M+1].

Example 4: N-((4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl)-2-((R)-9-(pyridine-2 -Yl)-6-oxaspiro[4.5]dec-9-yl)ethylamine (H-4)

Step 1: Dissolve 2-(thiophen-3-yl)ethanol (1g, 7.25mmol) in 1,4-dioxane (15ml), add 2,2-dimethoxyethylamine (1.17g, 10.87 mmol) and sulfuric acid (1.7ml), stirring at room temperature for 2 hours. The organic phase was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to obtain (4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)carboxamide (20 mg) as Rate: 1%. MS m/z(ESI): 170.1[M+1].

Step 2: Dissolve (4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)carboxamide (20mg, 0.118mmol) in chloroform (4ml) solution and add (R)-2-(9-(pyridin-2-yl)-6-oxospiro[4.5]dec-9-yl)acetaldehyde (31mg, 0.118mmol), trifluoroacetic acid (1 drop) and cyano Sodium borohydride (20mg, 0.236mmol) was stirred at room temperature and reacted overnight. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to obtain the target product N-((4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl) -2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]dec-9-yl)ethylamine H-4 (2.03 mg), the yield is 4%. MS m/z(ESI): 413.1[M+1].

Example 5: N-((4,4-Dimethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl)-2-((R )-9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine (H-5)

Step 1: Dissolve methyl 2-(thiophen-3-yl)acetate (1.56g, 0.01mol) in 40mL N,N-dimethylformamide, add methyl iodide (4.26g, 0.03mol), and cool to 2-5 degrees, add NaH (60%, 1.6 g, 0.04 mol) in batches, and stir at room temperature for 1 hour. 80mL of water was added to the reaction solution, extracted with ethyl acetate (30mL×3), the organic phases were combined, washed with saturated brine (40mL×1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and used a silica gel column The residue obtained was purified by chromatography with eluent (petroleum ether/ethyl acetate=4/1) to obtain the product methyl 2-methyl-2-(thiophen-3-yl)propionate (1.5g, yellow oily liquid) ), yield: 81.5%. MS m/z(ESI): 185.3[M+1].

Step 2: Slowly add methyl 2-methyl-2-(thiophen-3-yl)propionate (35ml) to the suspension of lithium tetrahydroaluminum (0.62g, 16.3mmol) in methyl tert-butyl ether (35ml) 1.5g, 8.15mmol) of methyl tert-butyl ether (10ml) solution, the mixture was stirred at room temperature for 1.5 hours, 10mL of ice water was added to the reaction solution, vigorously stirred for 5 minutes, filtered, separated, the organic phase was saturated with salt Wash with water (15mL×1), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the resulting residue by silica gel column chromatography with eluent (petroleum ether/ethyl acetate=2/1) to obtain product 2 -Methyl-2-(thiophen-3-yl)propan-1-ol (1 g, yellow oily liquid), yield: 78.7%. MS m/z(ESI): 138.3[M-18].

Step 3: Add 2-methyl-2-(thiophen-3-yl)propan-1-ol (1g, 6.4mmol) and 2,2-dimethoxyethylamine (0.74g, 7.04mmol) to dioxy Slowly add trifluoromethanesulfonic acid (2ml) solution to the six-ring (30ml) solution. The mixture was stirred at room temperature for 1.5 hours. 40mL of ice water was added to the reaction solution. The pH was adjusted to 9-10 with saturated potassium carbonate. Ester extraction (30ml*3), the combined organic phase was washed with saturated brine (15mL×1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the eluent (petroleum ether/acetic acid) Ethyl acetate = 2/1) Purify the resulting residue to obtain the product (4,4-dimethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methylamine (1.1g, yellow oily liquid), yield: 87.3%. MS m/z(ESI): 198.3[M+1].

Step 4: Combine (4,4-dimethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methylamine (60mg, 0.3mmol) and (R) -2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)acetaldehyde (78mg, 0.3mmol) was dissolved in 8mL 1,2-dichloroethane and added 0.5 mL of tetraisopropyl titanate was stirred at 45°C for 18 hours. After cooling to room temperature, sodium borohydride (34 mg, 0.9 mmol) was added to the reaction solution, stirred for 3 hours, 5 mL of water was added to the reaction solution, stirred for 0.5 minutes, filtered, the filtrate was concentrated under reduced pressure, and the residue obtained was purified by preparative chromatography to obtain the product N-((4,4-Dimethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl)-2-((R)-9- (Pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine H-5 (55 mg, brown solid), yield: 42%. MS m/z (ESI): 441.3 [M+1]. ¹ H NMR(400MHz,DMSO-d6)δ8.54–8.47(m,1H), 8.17(s,1H), 7.75–7.66(m,1H), 7.43(d,J=8.0Hz,1H), 7.30 (d,J=5.1Hz,1H), 7.22–7.14(m,1H), 6.98(d,J=5.1Hz,1H), 4.69(d,J=6.5Hz,1H), 3.64–3.52(m, 3H), 3.35–3.31(m,1H), 2.86–2.71(m,2H), 2.55-2.51(m,1H), 2.40-2.38(m,1H), 2.35–2.26(m,1H), 2.05( s, 1H), 1.98-1.84 (m, 1H), 1.75-1.60 (m, 3H), 1.58-1.27 (m, 6H), 1.17-1.06 (m, 6H), 0.93 (m, 1H), 0.59 ( m,1H).

Example 6: N-((4-ethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl)-2-((R)-9 -(Pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine (H-6)

Step 1: Dissolve methyl 2-(thiophen-3-yl)acetate (1.56g, 0.01mol) in 40mL N,N-dimethylformamide, add ethyl iodide (2.34g, 0.015mol), and cool At 2-5°C, NaH (60%, 1.6 g, 0.04 mol) was added in batches, and stirred at room temperature for 1 hour. 80mL of water was added to the reaction solution, extracted with ethyl acetate (30mL×3), the organic phases were combined, washed with saturated brine (40mL×1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and used a silica gel column The resulting residue was purified by chromatography with eluent (petroleum ether/ethyl acetate=4/1) to obtain methyl 2-(thiophen-3-yl)butyrate (1.1 g, yellow oily liquid), yield: 59.8 %. MS m/z(ESI): 185.3[M+1].

Step 2: Slowly add methyl 2-(thiophen-3-yl)butyrate (1.1g, 5.98mmol) to the suspension of lithium tetrahydroaluminum (0.45g, 11.96mmol) in methyl tert-butyl ether (35ml) ) In methyl tert-butyl ether (10ml), the mixture was stirred at room temperature for 1.5 hours, 10mL ice water was added to the reaction solution, stirred vigorously for 5 minutes, filtered, separated into layers, the organic phase was washed with saturated brine (15mL× 1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue obtained was purified by silica gel column chromatography with eluent (petroleum ether/ethyl acetate = 2/1) to obtain 2-(thiophen-3- Yl)butan-1-ol (0.65 g, yellow oily liquid), yield: 69.7%. MS m/z(ESI): 138.3[M-18].

Step 3: To 2-(thiophen-3-yl)butan-1-ol (0.65g, 4.2mmol) and 2,2-dimethoxyethylamine (0.48g, 4.58mmol) in dioxane (30ml ) Solution was slowly added trifluoromethanesulfonic acid (2ml) solution, the mixture was stirred at room temperature for 1.5 hours, 40mL ice water was added to the reaction solution, pH = 9-10 with saturated potassium carbonate, ethyl acetate extraction (30ml *3), the combined organic phase was washed with saturated brine (15mL×1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and silica gel column chromatography was used to determine the eluent (petroleum ether/ethyl acetate = 2 /1) Purify the resulting residue to obtain (4-ethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methylamine (0.56g, yellow oily liquid) , Yield: 67.7%. MS m/z(ESI): 198.3[M+1].

Step 4: Combine (4-ethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methylamine (59mg, 0.3mmol) and (R)-2- (9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)acetaldehyde (78mg, 0.3mmol) was dissolved in 8mL 1,2-dichloroethane and 0.5mL was added Tetraisopropyl titanate was stirred and reacted at 45°C for 18 hours. After cooling to room temperature, sodium borohydride (34 mg, 0.9 mmol) was added to the reaction solution, stirred for 3 hours, 5 mL of water was added to the reaction solution, stirred for 0.5 minutes, filtered, the filtrate was concentrated under reduced pressure, and the residue obtained was purified by preparative chromatography to obtain the product N-((4-ethyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl)-2-((R)-9-(pyridine- 2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine H-6 (21 mg, brown solid, formate), yield: 15.9%. MS m/z (ESI): 441.3 [M+1]. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.53–8.46(m,1H), 7.74–7.65(m,1H), 7.42(d,J=8.0Hz,1H), 7.30-7.26(m,1H) ), 7.20–7.12 (m, 1H), 6.91 (d, J = 5.1 Hz, 1H), 4.54 (t, J = 6.6 Hz, 1H), 4.02-3.96 (m, 1H), 3.56-3.50 (m, 3H), 3.30-3.26 (m, 1H), 2.69-2.65 (m, 2H), 2.55-2.52 (m, 1H), 2.40-2.37 (m, 2H), 2.31-2.28 (m, 1H), 1.97-- 1.79(m,2H), 1.79-1.22(m,11H), 0.98-0.79(m,4H), 0.64-0.51(m,1H).

Example 7-10

The preparation method of Example 7 refers to Example 6, except that the ethyl iodide in step 1 is replaced with methyl iodide.

The preparation method of Example 8 refers to Example 5, except that the methyl iodide in step 1 is replaced with ethyl iodide.

The preparation method of Example 9 refers to Example 6, except that the ethyl iodide in step 1 is replaced with 2-iodopropane.

The preparation method of Example 10 refers to Example 6, except that the ethyl iodide in step 1 is replaced with 1-iodopropane.

Example 11: N-(4-cyclobutyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl)-2-((R)-9 -(Pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine (H-11)

Step 1: Dissolve methyl 2-(thiophen-3-yl)acetate (500mg, 3.20mmol) in 10mL of dimethylformamide, add potassium tert-butoxide (539mg, 4.80mmol) at 0°C, and stir for reaction 30 Then, bromocyclobutane (475 mg, 3.52 mmol) was added, and the reaction was stirred at room temperature for 2 hours. 80 mL of ethyl acetate was added to the reaction solution, and washed with saturated sodium chloride solution (30 mL×3) and water (30 mL) in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent (petroleum ether/ethyl acetate = 1/0 ~ 4/1) to obtain 2-ring Butyl-2-(thiophen-3-yl) methyl acetate (498 mg, colorless oil), yield: 74.0%. MS m/z(ESI): 211.1[M+1].

Step 2: Dissolve methyl 2-cyclobutyl-2-(thiophen-3-yl)acetate (498mg, 2.37mmol) in 10mL of tetrahydrofuran, cool to 0°C under nitrogen protection, and add lithium aluminum hydride (135mg, 3.56 mmol), the reaction was stirred at 0°C for 1 hour. The reaction solution was poured into 70 mL saturated ammonium chloride solution for quenching, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent (petroleum ether/ethyl acetate = 1/0 ~ 4/1) to obtain 2- Cyclobutyl-2-(thiophen-3-yl)ethanol (407 mg, colorless oil), yield: 94.4%. MS m/z(ESI): 165.1[M-17].

Step 3: Dissolve 2-cyclobutyl-2-(thiophen-3-yl)ethanol (205mg, 1.12mmol) and aminoacetaldehyde dimethylacetal (176mg, 1.67mmol) in 5mL 1,4-dioxane Add 0.5 mL of trifluoromethanesulfonic acid while cooling under the protection of nitrogen, and stir and react at room temperature for 1 hour. After neutralization with potassium hydroxide aqueous solution, 30 mL of saturated sodium bicarbonate solution was added and extracted with ethyl acetate (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain (4-cyclobutyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl) Methylamine (205 mg, yellow oil), yield: 81.6%. MS m/z(ESI): 224.1[M+1].

Step 4: Combine (R)-2-(9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)acetaldehyde (50mg, 0.193mmol) and (4-cyclobutane 4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methylamine (47 mg, 0.21 mmol) was dissolved in 10 mL of methanol, a drop of acetic acid was added, and the reaction was stirred for 30 minutes. Sodium cyanoborohydride (49 mg, 0.78 mmol) was added to the reaction solution, and the reaction was stirred overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by preparative chromatography to obtain N-(4-cyclobutyl-4,7-dihydro-5H-thieno[2,3-c]pyran-7-yl)methyl )-2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)ethylamine H-11 (33.74mg, pale yellow oil), produced Rate: 37%. MS m/z (ESI): 467.3 [M+1]. ¹ H NMR(400MHz,CD ₃ OD)δ8.51(d,J=4.8Hz,1H),7.81-7.65(m,1H),7.48(d,J=8.1Hz,1H),7.23-7.16(m ,2H),6.88–6.76(m,1H), 4.74–4.56(m,1H),4.01–3.54(m,4H), 2.78–2.28(m,7H),2.22–1.32(m,17H),1.11 -1.05 (m, 1H), 0.74-0.67 (m, 1H).

Example 12: N-((2-methyl-1,2,3,4-tetrahydroisoquinolin-1-yl)methyl)-2-((R)-9-(pyridin-2-yl )-6-oxaspiro[4.5]dec-9-yl)ethylamine (H-12)

Step 1: Dissolve 2-(1,3-dioxoisoindolin-2-yl)acetic acid (10g, 48mmol) in 80mL tetrahydrofuran, add oxalyl chloride (6.2mL, 73mmol) in sequence at 0°C to catalyze The amount of N,N-dimethylformamide. The reaction was stirred at 0°C for 2 hours. Concentrated under reduced pressure to obtain the product 2-(1,3-dioxoisoindolin-2-yl)acetyl chloride (10.7 g, yellow liquid), yield: 100%. MS m/z(ESI): 224.0[M+1] (quenched with methanol, sample delivery).

Step 2: 2-(1,3-dioxoisoindolin-2-yl)acetyl chloride (10.7g, 48mmol) was added to 100mL of anhydrous dichloromethane, and 2-phenyl- 1-Ethylamine (5.8 g, 48 mmol), triethylamine (13.4 mL, 96 mmol). The reaction was stirred at room temperature for 2 hours. 50mL of water was added to the reaction solution, extracted with dichloromethane (100mL×2), the organic phases were combined, washed with water (50mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and eluted by silica gel column chromatography Reagent system (petroleum ether: ethyl acetate: 10/1) to purify the obtained residue to obtain 2-(1,3-dioxoisoindolin-2-yl)-N-phenethylacetamide in yield : 81%. MS m/z(ESI): 309.1[M+1].

Step 3: Dissolve 2-(1,3-dioxoisoindolin-2-yl)-N-phenethylacetamide (8g, 259mmol) in 80mL acetonitrile, add phosphorus oxychloride (7.25mL , 778mmol), the reaction was stirred at 80°C for 80 hours. Concentrate under reduced pressure, cool the concentrate to room temperature, dilute with dichloromethane (200mL), wash with water (50mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain 2-((3,4-dihydroisoquinoline) -1-yl)methyl)isoindole-1,3-dione (7.5g, yellow oil), yield: 99.8%, the crude product was directly used in the next reaction. MS m/z(ESI): 291.1[M+1].

Step 4: Dissolve 2-((3,4-dihydroisoquinolin-1-yl)methyl)isoindole-1,3-dione (1.45mg, 50mmol) in 15mL dichloromethane and add Sodium triacetylborohydride (3.18g, 150mmol), catalytic amount of acetic acid, stirred at room temperature for 3 hours. 20mL of water was added to the reaction solution, extracted with dichloromethane (80mL×2), the organic phases were combined, washed with water (20mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the product 2-((1, 2,3,4-Tetrahydroisoquinolin-1-yl)methyl)isoindole-1,3-dione, yield: 97.2%, the crude product was directly used in the next reaction. MS m/z(ESI): 293.1[M+1].

Step 5: 2-((1,2,3,4-tetrahydroisoquinolin-1-yl)methyl)isoindole-1,3-dione (1.4g, 4.78mmol) was added to 10mL methanol , Add 2-phenyl-1-amine (0.29g, 9.6mmol), catalytic amount of acetic acid. The reaction was stirred at 80°C for 2 hours. 20mL of water was added to the reaction solution, extracted with dichloromethane (40mL×2), the organic phases were combined, washed with water (50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 2-((2-methyl 1,2,3,4-tetrahydroisoquin-1-yl)methyl)isoindole-1,3-dione (0.7g, yellow oil), yield: 48.6%. MS m/z(ESI): 307.1[M+1].

Step 6: 2-((2-Methyl-1,2,3,4-tetrahydroisoquin-1-yl)methyl)isoindole-1,3-dione (0.6g, 1.96mmol ) Was added to 5 mL of ethanol, and hydrazine hydrate (0.39 g, 7.84 mmol) was added. The reaction was stirred at 80°C for 2 hours. Add 5 mL of potassium hydroxide aqueous solution to the reaction solution, extract with dichloromethane/MeOH (15/1) (32 mL×2), combine the organic phases, wash with water (20 mL), dry with anhydrous sodium sulfate, filter, and depressurize the filtrate Concentrated to obtain the product (2-methyl-1,2,3,4-tetrahydroisoquinolin-1-yl)methylamine (0.28 g, yellow oil), yield: 81.1%. MS m/z(ESI): 177.1[M+1].

Step 7: Dissolve (2-methyl-1,2,3,4-tetrahydroisoquinolin-1-yl)methylamine (78mg, 0.3mmol) in 5mL methanol, add (R)-2-( 9-(pyridin-2-yl)-6-oxaspiro[4.5]decane-9-yl)acetaldehyde (54mg, 0.3mmol), sodium cyanoborohydride (95mg, 1.5mmol), and a catalytic amount of acetic acid. The reaction was stirred at 70°C for 3 hours. 20mL of water was added to the reaction solution, filtered, the filtrate was extracted with dichloromethane (20mL×2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by preparative chromatography to obtain N-((2-methyl- 1,2,3,4-Tetrahydroisoquin-1-yl)methyl)-2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]dec-9- Yl)ethylamine H-12 (1.24 mg, yellow solid), yield: 4.96%. MS m/z (ESI): 420.2 [M+1]. ¹ H NMR(400MHz,CD ₃ OD)δ8.59-8.48(m,1H),8.35(brs,1H),7.78(t,J=7.8Hz,1H),7.48(d,J=8.1Hz,1H ), 7.25 (dd, J = 7.1, 5.3 Hz, 1H), 7.20-7.14 (m, 2H), 7.10 (s, 1H), 3.75-3.68 (m, 3H), 3.21-3.03 (m, 3H), 2.99–2.84(m,2H), 2.83–2.72(m,1H), 2.63–2.53(m,1H), 2.50–2.32(m,5H), 2.16–2.06(m,1H), 1.94–1.82(m ,2H),1.72-1.32(m,8H),1.11-1.02(m,1H),0.73-0.60(m,1H).

Examples 13 to 21

The preparation method of Example 13 refers to Example 4, except that the 2,2-dimethoxyethylamine in step 1 is replaced with 1,1-dimethoxy-2-propylamine.

The preparation method of Example 14 refers to Example 4, except that the 2,2-dimethoxyethylamine in step 1 is replaced with 1,1-dimethoxy-2-methylpropane-2-amine.

The preparation method of Example 15 refers to Example 4, except that the 2,2-dimethoxyethylamine in step 1 is replaced with 2,2-dimethoxypropane-1-amine.

The preparation method of Example 16 refers to Example 4, except that the 2-(thiophen-3-yl)ethanol in step 1 is replaced with 2-(thiophen-2-yl)ethanol.

The preparation method of Example 17 refers to Example 4, except that the 2-(thiophen-3-yl)ethanol in step 1 is replaced with 2-(furan-3-yl)ethanol.

Example 18

Step 1: Refer to Step 6 of Example 12, except that the raw material 2-((2-methyl-1,2,3,4-tetrahydroisoquin-1-yl)methyl)isoindoline The -1,3-dione was replaced with 2-((1,2,3,4-tetrahydroisoquinolin-1-yl)methyl)isoindole-1,3-dione.

Step 2: Refer to Step 7 of Example 12, except that (2-methyl-1,2,3,4-tetrahydroisoquinolin-1-yl)methylamine was replaced with (3,4-dihydro Isoquinolin-1-yl)methylamine.

Step 3: Refer to Step 4 of Example 12, except that 2-((3,4-dihydroisoquinolin-1-yl)methyl)isoindole-1,3-dione was replaced with (R )-N-((3,4-Dihydroisoquinolin-1-yl)methyl)-2(9-(pyridin-2-yl)-6-oxaspiro[4.5]dec-9-yl) -Ethylamine.

The preparation method of Example 19 refers to steps 2 to 4 of Example 11. The difference is that methyl 2-cyclobutyl-2-(thiophen-3-yl)acetate in step 2 is replaced with 2-dimethylamino- Ethyl 2-(thiophen-3-yl)acetate.

The preparation method of Example 20 refers to Example 6, except that the iodoethane in step 1 is replaced with 1-iodo-2-methoxyethane.

The preparation method of Example 21 refers to Example 6, except that the iodoethane in step 1 is replaced with 1,1'-oxybis[2-iodoethane].

Example 22: N-((7,7-Dimethyl-6,7-dihydro-4H-thiophene[3,2-c]pyran-4-yl)methyl)-2-((R) -9-(Pyridin-2-yl)-6-oxaspiro[4.5]dec-9-yl)ethane-1-amine (H-22)

Step 1: Dissolve methyl 2-(thiophen-2-yl)acetate (1.5g, 9.6mmol) in DMF (30ml), add sodium hydride (60%, 1.15g, 28.8mmol) at 0 degree, 0 degree After stirring for half an hour, methyl iodide (4.1 g, 28.8 mmol) was added, and the mixture was stirred at room temperature for 4 hours. Add ethyl acetate (100ml), wash with saturated sodium chloride solution, dry with anhydrous sodium sulfate, concentrate the organic phase under reduced pressure, and purify the obtained by thin layer chromatography with eluent (petroleum ether: ethyl acetate=85:15) From the residue, methyl 2-methyl-2-(thiophen-2-yl)propionate (1.3 g) was obtained in a yield of 73.4%. MS m/z(ESI): 185.1[M+1].

Step 2: Dissolve methyl 2-methyl-2-(thiophen-2-yl)propionate (300mg, 1.63mmol) in tetrahydrofuran (10ml), add tetrahydrolithium aluminum (186mg, 4.89mmol) at 0°C , Stir at 0 degrees for 1 hour. Sodium sulfate decahydrate (200 mg) was added, stirred at room temperature for half an hour, filtered, and the organic phase was concentrated under reduced pressure to obtain 2-methyl-2-(thiophen-2-yl)propanol (210 mg) with a yield of 82.3%. MS m/z(ESI): 157.1[M+1].

Step 3: Dissolve 2-methyl-2-(thiophen-2-yl)propanol (60mg, 0.38mmol) in 1,4-dioxane (5ml) and add 2,2-dimethoxyethane Amine (62mg, 0.58mmol) and trifluoromethanesulfonic acid (0.2ml) were stirred at room temperature for 2 hours. The organic phase was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to obtain the crude compound (7,7-dimethyl-6,7-dihydro-4H-thiophene[3,2-c]pyran-4- Yl) formamide (60 mg). MS m/z(ESI): 198.1[M+1].

Step 4: Dissolve (7,7-dimethyl-6,7-dihydro-4H-thiophene[3,2-c]pyran-4-yl)carboxamide (60mg, 0.3mmol) in chloroform (6ml) solution, add (R)-2-(9-(pyridin-2-yl)-6-oxospiro[4.5]dec-9-yl)acetaldehyde (78mg, 0.3mmol), trifluoroacetic acid (1 drop) and sodium cyanoborohydride (57mg, 0.9mmol), stirred at room temperature overnight. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to obtain N-((7,7-dimethyl-6,7-dihydro-4H-thiophene[3,2-c]pyran-4- (Yl)methyl)-2-((R)-9-(pyridin-2-yl)-6-oxaspiro[4.5]dec-9-yl)ethane-1-amine H-22 (16mg), The yield is 4%. MS m/z(ESI): 441.3[M+1]; ¹ H NMR(400MHz, DMSO-d6): δ8.51(s,1H), 8.21(s,1H), 7.72-7.70(m,1H) ,7.42-7.40(m,1H),7.28-7.26(m,1H),7.20-7.18(m,1H),6.78-6.76(m,1H),4.61-4.59(m,1H),3.58-3.34( m,7H),2.94-2.92(m,1H),2.65-2.63(m,1H),2.30-2.28(m,2H),1.77-1.62(m,4H),1.58-1.30(m,5H), 1.18 (s, 3H), 1.15 (s, 3H), 0.94-0.92 (m, 1H), 0.64-0.62 (m, 1H).

The preparation method of Example 23 refers to Example 4, except that the 2-(thiophen-3-yl)ethanol in step 1 is replaced with 2-(5-chloro-thiophen-3-yl)ethanol.

The preparation method of Example 24 refers to Example 1, except that the methyl 1-(thiophen-3-yl)cyclopentanecarboxylate in step 2 is replaced with 2-(5-fluoro-thiophen-3-yl)acetic acid Methyl ester.

The preparation method of Example 25 refers to Example 4, except that the 2-(thiophen-3-yl)ethanol in step 1 is replaced with 2-(5-methylthiophen-3-yl)ethanol.

The preparation method of Example 26 refers to Example 4, except that the 2-(thiophen-3-yl)ethanol in step 1 is replaced with 2-(4-methylthiophen-3-yl)ethanol.

The preparation method of Example 27 refers to Example 4, except that the 2-(thiophen-3-yl)ethanol in step 1 is replaced with 2-(4-chloro-thiophen-3-yl)ethanol.

The preparation method of Example 28 refers to Example 4, except that the 2-(thiophen-3-yl)ethanol in step 1 is replaced with 2-phenyl-1-ol.

Example 29

Step 1-2: Refer to Steps 2 and 3 of Example 12, except that 2-phenyl-1-ethylamine is replaced with 2-thiophen-1-ethylamine.

Step 3: Refer to Step 6 of Example 12, except that the raw material 2-((2-methyl-1,2,3,4-tetrahydroisoquin-1-yl)methyl)isoindoline The -1,3-dione was replaced with 2-((1,2,3,4-tetrahydroisoquinolin-1-yl)methyl)isoindole-1,3-dione.

Step 4: Refer to Step 7 of Example 12, except that (2-methyl-1,2,3,4-tetrahydroisoquinolin-1-yl)methylamine is replaced with (6,7-dihydro Thieno[3,2-c]pyridin-4-yl)methylamine.

Step 5: Refer to Step 4 of Example 12, except that 2-((3,4-dihydroisoquinolin-1-yl)methyl)isoindole-1,3-dione was replaced with (R )-N-(((6,7-Dihydrothieno[3,2-c]pyridin-4-yl)methyl)-2(9-(pyridin-2-yl)-6-oxaspiro[ 4.5] Dec-9-yl)-ethylamine.

The preparation method of Example 30 refers to Example 12, except that the 2-phenyl-1-amine in step 2 is replaced with 2-(thiophen-2-yl)-ethylamine.

Example 31

Step 1-2: Refer to Steps 2 and 3 of Example 12, except that 2-phenyl-1-ethylamine is replaced with 2-thiophene-3-ethylamine.

Step 3: Refer to Step 6 of Example 12, except that the raw material 2-((2-methyl-1,2,3,4-tetrahydroisoquin-1-yl)methyl)isoindoline The -1,3-dione was replaced with 2-((4,5-dihydrothieno[2,3-c]pyridin-7-yl)methyl)isoindole-1,3-dione.

Step 4: Refer to Step 7 of Example 12, except that (2-methyl-1,2,3,4-tetrahydroisoquinolin-1-yl)methylamine is replaced with (4,5-dihydro Thieno[2,3-c]pyridin-7-yl)methylamine.

Step 5: Refer to Step 4 of Example 12, except that 2-((3,4-dihydroisoquinolin-1-yl)methyl)isoindole-1,3-dione was replaced with (R )-N-(((4,5-Dihydrothieno[2,3-c]pyridin-7-yl)methyl)-2(9-(pyridin-2-yl)-6-oxaspiro[ 4.5] Dec-9-yl)-ethylamine.

The preparation method of Example 32 refers to Example 12, except that the 2-phenyl-1-amine in step 2 is replaced with 2-(thiophen-3-yl)-ethylamine.

The preparation method of Example 33 refers to Example 1, except that the 1,4-dibromobutane in step 1 is replaced with 1,2-dibromoethane.

The preparation method of Example 34 was prepared by referring to the above method.

The preparation method of Example 35 refers to Example 1, except that the 1,4-dibromobutane in step 1 is replaced with 1,3-dibromopropane, and the methyl 2-(thiophen-3-yl)acetate is replaced with Methyl 2-(thiophen-2-yl)acetate.

The preparation method of Example 36 refers to Example 6, except that the ethyl iodide in step 1 is replaced with 2-iodopropane, and methyl 2-(thiophen-3-yl)acetate is replaced with 2-(thiophen-2- Base) methyl acetate.

The preparation method of Example 37 refers to Example 1, except that the 1,4-dibromobutane in step 1 is replaced with 1,2-dibromoethane, and the methyl 2-(thiophen-3-yl)acetate is replaced with Into 2-(thiophen-2-yl) methyl acetate.

Biological test

The cell line used in the following test example is

CHO-K1OPRM1β-Arrestin Cell Line, source: DiscoverX, number: 93-0213C2, batch number: 13K0402.

The reagents used, their suppliers, article numbers and storage temperatures are as follows:

Assay Complete ^TM Cell Culture Kit 107, DiscoverX, 92-3107G, -20℃;

AssayComplete ^TM Thawing Reagent, DiscoverX, 92-4002TR, -20℃;

AssayComplete ^TM Cell Detachment Reagent, DiscoverX, 92-0009, -20℃;

Assay Complete ^TM Cell Plating Reagent, DiscoverX, 93-0563R2, -20℃;

Pathhunter Detection Kit, DiscoverX, 93-0001, -20℃;

PBS(1×)0.0067M(PO4), Hyclone, SH30256.01, 4℃;

DMSO, Sigma, D5879-100ML, room temperature;

NKH477, Sigma, 1603, -20℃;

IBMX, Tocris, I5879, -20°C.

The instruments used, their models and suppliers are as follows:

Countsatr BioMed, IM1200, ALIT;

Microscope, IX51, OLYMPUS;

Centrifuge, 5804, Eppendorf;

Thermostatic Water Bath, DK-S420, ShanghaiShenxian thermostatic equipment factory;

Cell Incubator, 3111, Thermo;

Biological Safety Cabinet, BSC-1300IIA2, AIRTECH;

OptiPlate-384White Opaque, 6007290, Perkin Elmer;

Multimode plate Reader, Victor X5, PerkinElmer;

Culture Plate-384White Opaque, TC-treated, 6007680, PerkinElmer.

Test case one HTRF-cAMP cell experiment

Experimental methods and procedures

1. Cell Recovery

1. Take the resuscitation solution out of the refrigerator at 4°C and place it in a water bath at 37°C for 15 minutes.

2. Take out the P6 generation cells from the liquid nitrogen tank, and quickly place the frozen cell cryopreservation tube in a 37°C water bath and gently shake for 30 seconds to 1 minute until small ice crystals or the cells are about to be completely melted.

3. Thoroughly disinfect and wipe dry with 70% alcohol.

4. Centrifuge to remove the cryopreserved fluid, and resuspend the cells in pre-warmed fresh resuscitation fluid.

a. Pipette 3ml of pre-warmed cell resuscitation solution into a 15ml centrifuge tube.

b. Centrifuge at 1300 rpm for 3 minutes.

c. Remove the supernatant cryopreservation solution and resuspend the cells with 4ml of pre-warmed resuscitation solution.

5. Transfer the cell suspension to a T25 cell culture flask for 24 hours, 37°C, 5% CO2.

6. After culturing for 24 hours, replace the resuscitation fluid in the cell culture flask with a pre-warmed cell culture medium.

Second, cell passaging

1. When the growth density of the cells in the T25 culture flask is >70%, use the cell digestion solution to digest and subculture the cells.

a. Aspirate the medium in the culture flask, add 4ml of pre-warmed PBS, gently shake to wash the cells, and aspirate the PBS.

b. Pipette 1ml of cell digestion solution into T25 culture flask.

c. Shake the culture flask repeatedly to completely cover the culture flask with digestive solution, and place it in a 37°C, 5% CO2 incubator for 5 minutes.

d. Take out the cell culture flask and observe the cells under the microscope to see if the cells are separated.

e. Add 3ml of pre-warmed cell culture medium to terminate the digestion.

f. Rinse the culture flask gently with cell culture medium repeatedly, and collect the cell suspension into a 15ml centrifuge tube.

g. Centrifuge at 1300 rpm for 3 minutes and remove the supernatant.

h. Resuspend with 3ml cell culture medium.

2. Passage the cells in a ratio of 1:3 (add 1ml of cell resuspension + 3ml of cell culture medium to each bottle and transfer to the T25 bottle).

Three, cell seed plate

1. Repeat steps 2.2.1 (a-h) until the cells reach the P8 generation. Count the cells and resuspend the cells with 2×/1mM IBMX stimulation buffer to make the cell density 1.2*10^6/ml.

2. Using a multi-channel pipette, plant the cell solution of 1.2*10^6/ml in a 384-well plate with a volume of 10ul per well (ie 12000 cells per well).

Four, c-AMP test

1. Configure related reagents and configure compounds according to the drug dilution configuration table.

a. 1×Stimulation buffer: Take 1ml of 5×Stimulation buffer and add it to 4ml of distilled water and mix well.

b. 2×/1mM IBMX stimulation buffer solution 5ml: Take 10ul 500mM IBMX storage solution and add it to 4990ul cell culture medium, gently pipette to mix.

c. Configuration table of gradient dilution of positive drug morphine:

d. Before the compound is diluted, dissolve the compound with DMSO to make the storage concentration 10mM.

The positive drug TRV130 and each compound dilution configuration table:

e. 50uM NK477 1ml: Take 1ul 50mM NKH477 storage solution and add it to 999ul 1×Stimulation buffer solution, shake and mix well.

f. Detection reagents

A. cAMP-Cryptate (donor, lyophilized) reaction solution: Take 1ml 5×cAMP-Cryptate stock solution and add it to 4ml 1×Lysis&Detection Buffer solution, and mix gently.

B. Anti-cAMP-d2 (acceptor, lyophilized) reaction solution: Take 1ml 5×Anti-cAMP-d2 storage solution and add it to 4ml 1×Lysis&Detection Buffer solution, and mix gently.

2. cAMP test procedure

a. Seed 12,000 cells in 10μl containing 2xIBMX stimulation buffer, per well.

b. Add 8 μl of compound sample dilution to each well of cells.

c. Add 2μl 10xNKH477 solution to each well.

d. Incubate at 37°C for 45mins.

e. Add 10μl cAMP-d2 and 10μl anti-cAMP Cryptate reaction solution.

f. Incubate for 60mins in the dark at room temperature.

g, HTRF plate reading.

3. RFU detection and reading

After 60 minutes of incubation, all samples will be detected and read by a homogeneous time-resolved fluorescence method.

data analysis

Export the data from the corresponding software in the computer connected to the multi-function plate reader, including two signal values of 665nm and 620nm. The calculation formula of the ratio is: ratio=665nm signal value/620nm signal value×10000. Use GraphPad Prism software to analyze the data. The best fit curve is log(agonist) vs. response. The EC50 value of the compound is determined by the non-linear regression analysis method of the computer-assisted dose-response curve; PEC50=-logEC50 (the unit of the EC50 value is mole); the maximum of% morphine Effect value = (compound sample ratio-blank hole ratio)/TOP×100 (Note: TOP value is the morphine sample ratio-blank hole ratio and then analyzed and fitted the curve Top value through the software Graphpad Prism). The test results of the example compounds are shown in Table 1:

Table 1 The activity of compounds on cAMP

Test Example 2 β-Arrestin cell experiment

Experimental methods and procedures

1. Cell Recovery

2. Take out the P6 generation cells from the liquid nitrogen tank, quickly place the frozen cell culture tube in a 37°C water bath and gently shake for 30 seconds to 1 minute until you see small ice crystals or the cells are about to melt completely.

3. Thoroughly disinfect and wipe dry with 70% alcohol.

b. Centrifuge at 1300 rpm for 3 minutes.

c. Remove the supernatant and resuspend the cells with 4ml of pre-warmed resuscitation fluid.

Second, cell passaging

b. Pipette 1ml cell digestion solution into T25 culture flask.

e. Add 3ml of pre-warmed cell culture medium to stop the digestion.

f. Wash the culture flask repeatedly with cell culture medium, and finally transfer the cell suspension to a 15ml centrifuge tube.

g. Centrifuge at 1300 rpm for 3 minutes and remove the supernatant.

h. Resuspend with 3ml cell culture medium.

3. Repeat steps 2.2.1 (a-h) until the cells reach the P8 generation.

Three, cell seed plate

1. Take 20ul of cell suspension with a pipette and measure the number of cells with a cell counter.

2. Centrifuge at 1300 rpm for 3 minutes to pellet the cells.

3. Remove the supernatant and add the corresponding cell plating solution to make the cell concentration 2×10^5/ml.

4. Using a multi-channel pipette, according to the experimental design, plant 2×10^5/ml cell solution in a 384-well plate with a volume of 20ul per well (ie 4000 cells per well).

5. Place the 384-well plate with the seeded cells in a 37°C, 5% CO2 incubator for 24 hours.

Four, β-arrestin test

1. Prepare the compound according to the following dilution table.

a. Configuration table of gradient dilution of positive drug morphine:

b. Before the compound is diluted, the compound is dissolved in DMSO to a storage concentration of 10mM.

The positive drug TRV130 and each compound dilution configuration table:

2. Take 5ul of each compound sample dilution prepared above and add it to a 384-well plate.

3. After adding the sample, put the 384-well plate back into the 37°C, 5% CO2 incubator and incubate for 90 minutes.

Five, RLU detection

1. Before the completion of the compound incubation, configure the Working Detection solution according to the following ratio (be careful to avoid light). Then add 12.5ul to each well, and incubate for 1h in a shaker at room temperature in the dark.

2. At the end of the compound incubation, add 12.5ul of the above working solution to each well, and incubate for 1 hour in the dark, room temperature, 80rpm shaker.

3. At the end of the incubation, use a multi-function plate reader to read the plate.

data analysis

Export the data from the corresponding software in the computer connected to the multi-function plate reader, and analyze the data with GraphPad Prism software. The best fit curve is log(agonist) vs. response. The EC50 value of the compound is determined by the non-linear regression analysis method of the computer-assisted dose-response curve; PEC50=-logEC50 (the unit of the EC50 value is mole); the maximum of% morphine Effect value = (RLU value of the compound sample-RLU value of the blank well)/TOP×100 (Note: TOP value is the RLU value of the morphine sample-the RLU value of the blank well, after analyzing the fitted curve Top value by the software Graphpad Prism) . The test results of the example compounds are shown in Table 2:

Table 2 Test results of compounds on β-arrestin

It can be seen from Table 1 and Table 2 that the representative compound of the present invention has a higher inhibitory activity on cAMP and a higher Emax value. In addition, the compound of the present invention has a lower Emax value for β-arrestin, and has a good bias.

All documents mentioned in the present invention are cited as references in this application, as if each document was individually cited as a reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

A compound represented by formula (I), or a pharmaceutically acceptable salt, stereoisomer or solvate thereof:

Where

R 0 is hydrogen or substituted or unsubstituted C 1-10 alkyl;

R 1 and R 2 are each independently hydrogen, halogen, substituted or unsubstituted C 1-10 alkyl;

Alternatively, R 1 , R 2 and the connected carbon atoms together form a 3 to 6 membered saturated or unsaturated monocyclic ring or a 3 to 6 membered saturated or unsaturated monocyclic ring; the 3 to 6 membered saturated or unsaturated monocyclic ring , 3 to 6-membered saturated or unsaturated monocyclic rings are unsubstituted or substituted by 1-3 substituents selected from the group consisting of halogen, C 1-10 alkoxy, C 1-10 alkyl, halogenated C 1-10 alkyl;

R 3 and R 4 are each independently hydrogen, hydroxyl, cyano, halogen, substituted or unsubstituted C 1-10 alkyl, substituted or unsubstituted C 1-10 alkoxy, halogenated C 1-10 alkane Group, substituted or unsubstituted C 3-8 cycloalkyl, NR 11 R 12 , or substituted or unsubstituted 4 to 6-membered saturated monocyclic heterocyclic ring;

Or R 3 , R 4 and the connected carbon atoms together form a 3 to 6 membered saturated or unsaturated monocyclic ring or a 3 to 6 membered saturated or unsaturated monocyclic ring; the 3 to 6 membered saturated or unsaturated monocyclic ring, The 3- to 6-membered saturated or unsaturated monocyclic ring is unsubstituted or substituted with 1-3 substituents selected from the group consisting of halogen, C 1-10 alkoxy, C 1-10 alkyl, halogenated C 1 -10 alkyl;

R 11 and R 12 are each independently hydrogen, substituted or unsubstituted C 1-10 alkyl, halo C 1-10 alkyl, substituted or unsubstituted C 3-8 cycloalkyl, or substituted or unsubstituted A 3 to 6-membered saturated or unsaturated monocyclic heterocyclic ring; or R 11 , R 12 and the connected nitrogen atom form a substituted or unsubstituted 4- to 6-membered saturated or unsaturated monocyclic heterocyclic ring;

X is O or NR c ; R c is hydrogen or C 1-10 alkyl;

R a , R b and the connected carbon atoms together form a substituted or unsubstituted C 6-10 aromatic ring, or a substituted or unsubstituted 5 or 6-membered monocyclic heteroaromatic ring, the C 6-10 aromatic ring or 5 Or a 6-membered monocyclic heteroaromatic ring and the connected heterocyclic ring form a fused bicyclic ring;

The "substitution" means that 1, 2, or 3 hydrogen atoms in the group are replaced by substituents each independently selected from Group A;

The group A substituents are selected from: cyano, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, halogenated C 1-8 alkyl, halogen, nitro, C 6-10 aryl, 5 or 6-membered monocyclic heteroaryl, C 1-10 alkyl, C 1-10 alkoxy, C 3-8 cycloalkyl, C 3-8 cycloalkoxy, C 2-10 alkenyl, C 2- 10 alkynyl, NR a0 R b0 , -CONR a0 R b0 , -C(O)OC 1-10 alkyl, -CHO, -OC(O)C 1-10 alkyl, -SO 2 C 1-10 alkane Group, -SO 2 C 6-10 aryl, -COC 6-10 aryl, 4 to 6-membered saturated or unsaturated monocyclic ring or 4 to 6-membered saturated or unsaturated monocyclic ring, wherein the C 6-10 Aryl, 5- or 6-membered monocyclic heteroaryl, 4- to 6-membered saturated or unsaturated monocyclic heterocyclic ring, or 4- to 6-membered saturated or unsaturated monocyclic ring is unsubstituted or 1, 2 or 3 selected from acetyl Group, hydroxyl, cyano, halogen, C 1-3 alkyl, C 1-3 alkoxy, C 3-6 cycloalkyl, NR a0 R b0 substituents; R a0 , R b0 are each independently Hydrogen or C 1-3 alkyl.
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein R 1 and R 2 are each independently hydrogen or C 1-3 alkyl.
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the C 6-10 aromatic ring formed by Ra , R b and the connected carbon atom is Benzene ring.
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein Ra , Rb and the connected carbon atom together form a 5- or 6-membered monocyclic heterocycle The aromatic ring is selected from the following structures:

among them
The two ring atoms represented are adjacent pairs of atoms shared when fused with other rings; the 5- or 6-membered monocyclic heteroaromatic ring is unsubstituted or is independently selected from 1, 2 or 3 Substituents of group A are substituted.
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein Ra , Rb and the connected carbon atom together form a 5- or 6-membered monocyclic heterocycle The aromatic ring is thiophene or furan.
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein R 3 and R 4 are each independently hydrogen, C 1-3 alkyl, C 1 -3 Alkoxy substituted C 1-3 alkyl, C 3-6 cycloalkyl, NH 2 , NH(C 1-3 alkyl) or N(C 1-3 alkyl) 2 ;

Or R 3 , R 4 and the connected carbon atoms together form a 4- to 6-membered saturated monocyclic ring or a 3- to 6-membered saturated monocyclic ring; the 4- to 6-membered saturated monocyclic ring or a 3- to 6-membered saturated monocyclic ring is not Substituted or substituted with 1-3 substituents selected from the group consisting of halogen, C 1-3 alkoxy, C 1-3 alkyl, halo C 1-3 alkyl.
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein X is O or NR c ; R c is hydrogen or methyl.
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, wherein:
Selected from the following structures:
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, wherein:
Selected from the following structures:
The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, wherein the compound is selected from Table A, wherein the compound of Table A is selected from the following group:
A pharmaceutical composition comprising the compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof; and a pharmaceutically acceptable carrier.
The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the pharmaceutical composition according to claim 11 in the preparation of the prevention and/or treatment of MOR Receptor agonist-mediated drug use in related diseases.
The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the pharmaceutical composition according to claim 11 is used in the preparation of the prevention and/or treatment of pain Use in medicine for pain-related diseases.
The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the pharmaceutical composition according to claim 11 in the preparation of agonistic or antagonistic MOR receptor Use in medicine.