Heteroaryl derivative and application thereof

Abstract

The invention discloses a heteroaryl derivative and application thereof, and particularly discloses compounds as shown in formulas (I) and (V) or pharmaceutically acceptable salts thereof.

Description

Heteroaryl derivatives and their applications

[0001] The present invention claims the following priority:

[0002] 1) priority and the benefit of Chinese Patent Application No. 202311284906.6 filed with the State Intellectual Property Office of China on September 28, 2023, 2) priority and the benefit of Chinese Patent Application No. 202311410637.3 filed with the State Intellectual Property Office of China on October 26, 2023, 3) priority and the benefit of Chinese Patent Application No. 202311849240.4 filed with the State Intellectual Property Office of China on December 28, 2023, 4) priority and the benefit of Chinese Patent Application No. 202410539834.3 filed with the State Intellectual Property Office of China on April 29, 2024, and 5) priority and the benefit of Chinese Patent Application No. 202411320022.6 filed with the State Intellectual Property Office of China on September 20, 2024, the disclosures of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention relates to a class of heteroaryl derivatives and applications thereof, and specifically to compounds represented by formulas (I) and (V) or pharmaceutically acceptable salts thereof. Background Art

[0004] Voltage-gated Kv7 / KCNQ potassium channels are widely present in the nervous system and play a crucial role in regulating neural excitability. Research on Kv7 potassium channel openers offers promising new avenues for the development of new drugs to treat disorders associated with neural hyperexcitability, such as epilepsy and pain.

[0005] The Kv7 family consists of five subtypes. Kv7.2 and Kv7.3 (encoded by KCNQ2 and KCNQ3) are homologous subunits of ubiquitously expressed neuronal voltage-gated potassium channels. They are key subunits involved in neuronal signaling and in regulating the hyperexcitability of epilepsy. KCNQ2 and KCNQ3 are primarily distributed in the brain, with high expression in the cerebellar cortex, amygdala, caudate nucleus, and hippocampus. They form homotetramers and heterotetramers (M-type potassium channels), which form the molecular basis for the formation of the M-type current. Increased M-type currents can hyperpolarize the cell membrane, thereby reducing neuronal excitability and preventing the occurrence and propagation of action potential bursts, leading to seizures. Enhancing the open state of Kv7.2 / Kv7.3 channels in neurons promotes a hyperpolarized resting state, reducing the spikes of fast action potentials (i.e., burst discharges), stabilizing the resting membrane potential, and thus limiting neuronal excitability, which contributes to the antiepileptic drug's efficacy.

[0006] Retigabine has been clinically demonstrated to be highly effective as a KCNQ2 / 3 potassium channel opener for the treatment of convulsions and epilepsy. Therefore, the development of KCNQ2 / 3 openers has become a hot topic in the clinical treatment of neurological diseases such as epilepsy. Xenon Biopharmaceutical Company's XEN1101 has entered Phase 3 clinical trials for the treatment of focal epileptic seizures. Biohaven Biohaven's BHV-7000 has also completed Phase 1 clinical trials, demonstrating significant reductions in side effects and is a promising drug for the treatment of neurological conditions such as epilepsy and pain.

[0007] Summary of the Invention

[0008] The present invention provides compounds represented by formula (I) and (V) or pharmaceutically acceptable salts thereof,

[0009] in,

[0010] Ring A is selected from 9-10 membered bicyclic heteroaryl;

[0011] Ring B is fused with ring A, and ring B is selected from C 4-7 Cycloalkyl, C 4-7 Cycloalkenyl, 4-7 membered heterocycloalkyl and 4-7 membered heterocycloalkenyl;

[0012] Each R1 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, CF3, C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl, the C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R a replace;

[0013] R2 is selected from cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 Cycloalkenyl, the cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b replace;

[0014] R3 is selected from H and -C(O)C 1-4 alkyl;

[0015] R4 is selected from C 1-6 Alkyl, C1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, each independently optionally substituted by 1, 2, 3, 4 or 5 R c replace;

[0016] Each R5 is independently selected from H, F, Cl, Br, I, =O, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0017] Each R6 is independently selected from H, F, Cl, Br, I, CN, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, 3-7 membered heterocycloalkyl and phenyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, 4-7 membered heterocycloalkyl and phenyl are each independently optionally substituted with 1, 2 or 3 R 6a Replacement; each R a and R c Each independently selected from H, F, Cl, Br, I, OH, NH2, CN, CH3 and CF3;

[0018] Each R b Each independently selected from H, F, Cl, Br, I, =O, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-3 Alkoxy is each independently optionally substituted with 1, 2 or 3 F;

[0019] Each R 6a are independently selected from H, F, Cl, Br, I, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 Alkoxy is each independently optionally substituted with 1, 2, 3, 4 or 5 F;

[0020] m is selected from 1, 2, 3 and 4;

[0021] p and q are independently selected from 1, 2 and 3;

[0022] The condition is,

[0023] 1) When R2 is selected from phenyl and 5-6 membered heteroaryl, the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b When substituted, the structural fragment Selected from or,

[0024] 2) When R2 is selected from cyclobutyl, the cyclobutyl is optionally substituted by 1, 2 or 3 R b When substituted, m is selected from 2, 3 and 4.

[0025] In some technical solutions of the present invention, the above R a are independently selected from H, F and OH, and other variables are as defined in the present invention.

[0026] In some technical solutions of the present invention, the above R b are independently selected from H, F, =O, CH3 and OCF3, and other variables are as defined in the present invention.

[0027] In some technical solutions of the present invention, the above R c are independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0028] In some technical solutions of the present invention, the above R1 is independently selected from H, F, Cl, OH, NH2, CN, CF3, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl, and the CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl are independently optionally replaced by 1, 2 or 3 R a Substitution, other variables are as defined in the present invention.

[0029] In some technical solutions of the present invention, each of the above R1 is independently selected from H, F, CN, CF3, Other variables are as defined in the present invention.

[0030] In some technical solutions of the present invention, the above R2 is selected from C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl, the C 5-8 Bridged ring cycloalkyl, C 5-8Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0031] In some technical solutions of the present invention, the above R2 is selected from cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridyl, oxetanyl, azetidinyl, piperidinyl, piperazinyl, cyclopentenyl and cyclohexenyl, and the cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridyl, oxetanyl, azetidinyl, piperidinyl, piperazinyl, cyclopentenyl and cyclohexenyl are independently optionally replaced by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0032] In some technical solutions of the present invention, the above R2 is selected from and Other variables are as defined in the present invention.

[0033] In some technical solutions of the present invention, the above R2 is Other variables are as defined in the present invention.

[0034] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0035] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0036] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0037] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0038] In some technical solutions of the present invention, the above R4 is selected from CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, CH2C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, The CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, CH2C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, and are independently optionally replaced by 1, 2, 3, 4 or 5 R c Substitution, other variables are as defined in the present invention.

[0039] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0040] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0041] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0042] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0043] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0044] In some technical solutions of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0045] In some technical solutions of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0046] In some technical solutions of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0047] The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0048] in,

[0049] Each R1 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, CF3, C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl, the C 2-4 Alkyl, C 1-4 Alkoxy, C3-7 Cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R a replace;

[0050] R2 is selected from cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 Cycloalkenyl, the cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b replace;

[0051] R3 is selected from H and -C(O)C 1-4 alkyl;

[0052] R4 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, each independently optionally substituted by 1, 2, 3, 4 or 5 R c replace;

[0053] Each R a and R c Each independently selected from H, F, Cl, Br, I, OH, NH2, CN, CH3 and CF3;

[0054] Each R b Each independently selected from H, F, Cl, Br, I, =O, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-3 Alkoxy is each independently optionally substituted with 1, 2 or 3 F;

[0055] m is selected from 1, 2, 3 and 4;

[0056] The condition is,

[0057] 1) When R2 is selected from phenyl and 5-6 membered heteroaryl, the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b When substituted, the structural fragment Selected from or,

[0058] 2) When R2 is selected from cyclobutyl, the cyclobutyl is optionally substituted by 1, 2 or 3 R b When substituted, m is selected from 2, 3 and 4.

[0059] The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0060] in,

[0061] Each R1 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl, the C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R a replace;

[0062] R2 is selected from cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 Cycloalkenyl, the cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b replace;

[0063] R3 is selected from H and -C(O)C 1-4 alkyl;

[0064] R4 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, each independently optionally substituted by 1, 2, 3, 4 or 5 R c replace;

[0065] Each R a and R c Each independently selected from H, F, Cl, Br, I, OH, NH2, CN, CH3 and CF3;

[0066] Each Rb Each independently selected from H, F, Cl, Br, I, =O, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-3 Alkoxy is each independently optionally substituted with 1, 2 or 3 F;

[0067] m is selected from 1, 2, 3 and 4;

[0068] The condition is,

[0069] 1) When R2 is selected from phenyl and 5-6 membered heteroaryl, the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b When substituted, the structural fragment Selected from or,

[0070] 2) When R2 is selected from cyclobutyl, the cyclobutyl is optionally substituted by 1, 2 or 3 R b When substituted, m is selected from 2, 3 and 4.

[0071] The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0072] in,

[0073] Each R1 is independently selected from F, Cl, Br, I, OH, NH2, CN, C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl, the C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R a replace;

[0074] R2 is selected from cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl, the cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R b replace;

[0075] R3 is selected from H and -C(O)C 1-4 alkyl;

[0076] R4 is selected from C 1-6Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, each independently optionally substituted by 1, 2, 3, 4 or 5 R c replace;

[0077] Each R a and R c Each independently selected from F, Cl, Br, I, OH, NH2, CN, CH3 and CF3;

[0078] Each R b are independently selected from F, Cl, Br, I, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-3 Alkoxy is each independently optionally substituted with 1, 2 or 3 F;

[0079] m is selected from 1, 2, 3 and 4;

[0080] The condition is,

[0081] 1) When R2 is selected from phenyl and 5-6 membered heteroaryl, the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b When substituted, the structural fragment Selected from or,

[0082] 2) When R2 is selected from cyclobutyl, the cyclobutyl is optionally substituted by 1, 2 or 3 R b When substituted, m is selected from 2, 3 and 4.

[0083] In some technical solutions of the present invention, the above R a are independently selected from H, F and OH, and other variables are as defined in the present invention.

[0084] In some technical solutions of the present invention, the above R a are independently selected from F and OH, and other variables are as defined in the present invention.

[0085] In some technical solutions of the present invention, the above R b are independently selected from H, F, =O, CH3 and OCF3, and other variables are as defined in the present invention.

[0086] In some technical solutions of the present invention, the above R bare independently selected from F, CH3 and OCF3, and other variables are as defined in the present invention.

[0087] In some technical solutions of the present invention, the above R c are independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0088] In some technical solutions of the present invention, the above R c are independently selected from F and CH3, and other variables are as defined in the present invention.

[0089] In some technical solutions of the present invention, each of the above R1 is independently selected from H, F, Cl, OH, NH2, CN, CF3, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl, and the CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl are independently optionally replaced by 1, 2 or 3 R a Substitution, other variables are as defined in the present invention.

[0090] In some technical solutions of the present invention, each of the above R1 is independently selected from H, F, CN, CF3, Other variables are as defined in the present invention.

[0091] In some technical solutions of the present invention, each of the above R1 is independently selected from H, F, CN and CF3, and other variables are as defined in the present invention.

[0092] In some technical solutions of the present invention, each of the above R1 is independently selected from H, F, Cl, OH, NH2, CN, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl, and the CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl are independently optionally replaced by 1, 2 or 3 R a Substitution, other variables are as defined in the present invention.

[0093] In some technical solutions of the present invention, each of the above R1 is selected from H, F, CN, Other variables are as defined in the present invention.

[0094] In some technical solutions of the present invention, each of the above R1 is independently selected from F, Cl, OH, NH2, CN, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl, and the CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl are independently optionally replaced by 1, 2 or 3 R a Substitution, other variables are as defined in the present invention.

[0095] In some technical solutions of the present invention, each of the above R1 is selected from F, CN, Other variables are as defined in the present invention.

[0096] In some technical solutions of the present invention, the above R2 is selected from C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, 4-7 membered heterocycloalkyl and C 5-8 Cycloalkenyl, the C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, 4-7 membered heterocycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0097] In some technical solutions of the present invention, the above R2 is selected from C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl, the C 5- 8-bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0098] In some technical solutions of the present invention, the above R2 is selected from C 5-8 Bridged cycloalkyl and C 5-8 Cycloalkenyl, the C 5-8 Bridged cycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0099] In some technical solutions of the present invention, the above R2 is selected from C 5-8 Bridged ring cycloalkyl, the C 5-8 The bridged cycloalkyl group is optionally substituted by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0100] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0101] In some technical solutions of the present invention, the above R2 is selected from cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridyl, oxetanyl, azetidinyl, piperidinyl, piperazinyl, cyclopentenyl and cyclohexenyl, and the cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridyl, oxetanyl, azetidinyl, piperidinyl, piperazinyl, cyclopentenyl and cyclohexenyl are independently optionally replaced by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0102] In some technical solutions of the present invention, the above R2 is selected from cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridinyl, oxetanyl, azetidinyl, piperidinyl and piperazinyl, and the cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridinyl, oxetanyl, azetidinyl, piperidinyl and piperazinyl are independently optionally replaced by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0103] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0104] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0105] In some technical solutions of the present invention, the above R2 is selected from C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl, the C 5- 8-bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0106] In some technical solutions of the present invention, the above R2 is selected from Other variables are as defined in the present invention.

[0107] In some technical solutions of the present invention, the above R4 is selected from CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, and The CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, and are independently optionally replaced by 1, 2, 3, 4 or 5 R c Substitution, other variables are as defined in the present invention.

[0108] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0109] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0110] In some technical solutions of the present invention, the above R4 is selected from and Other variables are as defined in the present invention.

[0111] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention.

[0112] In some technical solutions of the present invention, the above R4 is selected from Other variables are as defined in the present invention. In some technical solutions of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0113] In some technical solutions of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0114] In some technical solutions of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0115] In some technical solutions of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0116] In some technical solutions of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from:

[0117] in,

[0118] Each R1 is independently selected from H, F, CN, CF3, R2 is selected from C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, 4-7 membered heterocycloalkyl and C 5-8 Cycloalkenyl, the C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, 4-7 membered heterocycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b replace;

[0119] R4 is selected from

[0120] In some technical solutions of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from:

[0121] in,

[0122] R2 is selected from phenyl, 5-6 membered heteroaryl and C 5-8 Cycloalkenyl, the phenyl, 5-6 membered heteroaryl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b replace;

[0123] Each R b , each R1 and R4 are as defined in the present invention.

[0124] In some technical solutions of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from:

[0125] in,

[0126] R2 is selected from C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl, the C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R b replace;

[0127] Each R1, R4, R b and m are as defined in the present invention.

[0128] In some technical solutions of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from:

[0129] wherein R2 and R4 are as defined in the present invention.

[0130] The present invention also provides a compound represented by formula (I') or a pharmaceutically acceptable salt thereof,

[0131] in,

[0132] Ring A is selected from 9-10 membered bicyclic heteroaryl;

[0133] Ring B is fused with ring A, and ring B is selected from C 4-7 Cycloalkyl, C 4-7 Cycloalkenyl, 4-7 membered heterocycloalkyl and 4-7 membered heterocycloalkenyl;

[0134] Each R5 is independently selected from H, F, Cl, Br, I, =O, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0135] Each R6 is independently selected from H, F, Cl, Br, I, CN, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, 3-7 membered heterocycloalkyl and phenyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, 4-7 membered heterocycloalkyl and phenyl are each independently optionally substituted with 1, 2 or 3 R 6a Substituted; R4 is selected from C 1-6 Alkyl, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl and -CH2-phenyl, the C 1-6 Alkyl, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl and -CH2-phenyl are each independently optionally substituted with 1, 2, 3, 4 or 5 Rc;

[0136] Each R 6a are independently selected from H, F, Cl, Br, I, OH, NH2, CN, C 1-4 Alkyl and C1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 Alkoxy is each independently optionally substituted with 1, 2, 3, 4 or 5 F;

[0137] Each Rc is independently selected from H, F, Cl, Br, I, CH3 and CF3;

[0138] m and n are independently selected from 1, 2 and 3, respectively.

[0139] In some technical solutions of the present invention, each of the above R5 is independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0140] In some technical solutions of the present invention, each of the above R6 is independently selected from H, F, Cl, CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl and phenyl, and the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl and phenyl are independently optionally replaced by 1, 2 or 3 R 6a Substitution, other variables are as defined in the present invention.

[0141] In some technical solutions of the present invention, each of the above R6 is independently selected from H, F, Cl, CH3, CF3, CH2CH3, C(CH3)3, CN, Other variables are as defined in the present invention.

[0142] In some technical solutions of the present invention, each of the above R6 is independently selected from H, F, Cl, CN, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl, and the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl are independently optionally replaced by 1, 2 or 3 R6 a Substitution, other variables are as defined in the present invention.

[0143] In some technical solutions of the present invention, each of the above R6 is independently selected from H, F, Cl, CH3, CF3, CH2CH3, C(CH3)3, CN, Other variables are as defined in the present invention.

[0144] In some technical solutions of the present invention, the above R4 is selected from CH2C(CH3)3, The CH2C(CH3)3, are each independently optionally substituted with 1, 2, 3, 4 or 5 Rc, and other variables are as defined herein.

[0145] In some technical solutions of the present invention, the above R4 is selected from CH2C(CH3)3, Other variables are as defined in the present invention.

[0146] In some technical solutions of the present invention, the ring A is selected from benzimidazolyl, benzopyrazolyl, pyridoimidazole, pyridopyrazolyl, pyrimidoimidazole, pyrimidopyrazolyl, indolyl and pyridazinoimidazole, and other variables are as defined in the present invention.

[0147] In some technical solutions of the present invention, the ring A is selected from benzimidazolyl, benzopyrazolyl, pyridoimidazolyl, pyridopyrazolyl, pyrimidoimidazolyl and pyrimidopyrazolyl, and other variables are as defined in the present invention.

[0148] In some technical solutions of the present invention, the ring B is selected from cyclopentyl, cyclohexyl, cyclopentenyl, cyclohexenyl, tetrahydropyrrolyl, dihydropyrrolyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, 1,3-dioxolyl and 1,4-dioxinyl, and other variables are as defined in the present invention.

[0149] In some technical solutions of the present invention, the ring B is selected from cyclopentyl, cyclohexyl, cyclopentenyl, cyclohexenyl, pyrrolidinyl and pyrrolidenyl, and other variables are as defined in the present invention.

[0150] In some technical solutions of the present invention, the above structural unit Selected from wherein R5, R6 and p are as defined in the present invention.

[0151] Some other solutions of the present invention are obtained by arbitrarily combining the above variables.

[0152] The present invention also provides the following compounds or pharmaceutically acceptable salts thereof:

[0153] The present invention also provides a pharmaceutical composition containing a therapeutically or prophylactically effective amount of the compound of the present invention, its stereoisomers and pharmaceutically acceptable salts thereof; further, the pharmaceutical composition also includes a pharmaceutically acceptable excipient.

[0154] The present invention also provides the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating diseases related to Kv7 potassium channel openers.

[0155] In some embodiments of the present invention, the above-mentioned Kv7 potassium channel opener-related diseases include but are not limited to epilepsy, inflammatory pain, neuropathic pain, migraine, depression, anxiety disorders, stroke, Alzheimer's disease, neurodegenerative diseases, neuronal hyperexcitability, complications caused by cocaine abuse, nicotine withdrawal syndrome, alcohol withdrawal syndrome and tinnitus, etc.

[0156] In some embodiments of the present invention, the above-mentioned Kv7 potassium ion channel includes Kv7.2, Kv7.3, Kv7.4 and Kv7.5; preferably Kv7.2 and Kv7.3.

[0157] The present invention also provides the following synthetic route:

[0158] Route 1:

[0159] Route 2:

[0160] Route 3:

[0161] Route 4:

[0162] Route 5:

[0163] Route 6:

[0164] Route 7:

[0165] The present invention also provides the following test method:

[0166] Test Method 1: hKCNQ2 / 3 Thallium Current Test

[0167] Aim: To determine the in vitro concentration-response relationship of compounds using FLIPR thallium flux analysis in a CHO cell line stably expressing hKCNQ2 / 3.

[0168] Experimental cells and reagents:

[0169] (1) Cell lines:

[0170] CHO cells stably transfected with hKCNQ2 / 3.

[0171] (2) Cell culture medium reagents:

[0172] Cell culture medium reagent information is shown in Table 1.

[0173] Table 1: Cell culture medium reagent information

[0174] (3) Cell culture:

[0175] This cell line is passaged three times weekly at a 1:2-1:3 ratio. When cells reach >80% confluency in a T-75 flask, use 0.25% trypsin-EDTA solution for approximately 1 minute and then transfer the cells from the flask for passage. Transfer the cells to another T-75 flask containing complete cell growth medium according to the passage ratio. Note: To maintain logarithmic cell growth, cells should be grown in a subconfluent monolayer. Passage cells every 2-3 days, depending on the cell doubling time.

[0176] Experimental process:

[0177] (1) Day 1, cell preparation: Cells were digested as described above, and cell density and viability were determined using a Cell Countess. The volume of the single-cell suspension was adjusted with complete cell growth medium. hKCNQ2 / 3_CHO cells were then seeded onto PDL-precoated 384-well plates at a density of approximately 20,000 cells / well (30 μL / well). The plates were then incubated overnight in a 37°C, 5% CO2 humidified air incubator.

[0178] (2) The next day, compound preparation: The test compound was diluted with dimethyl sulfoxide (DMSO) to the storage concentration and stored in a refrigerator at -20°C. The compound addition program was set on the ECHO liquid workstation and the compound was added to the plate using ECHO. Retigabine (RTG) was used as the positive reference, with the highest concentration being 100 μM, 3-fold dilution, and 9 doses as the plate map required for plate making. + and 1 mM Tl + Prepare plates with stimulation buffer.

[0179] Note: 1) K2SO4 and Tl2SO4 are prepared in 1× chloride-free buffer. When different voltage-gated potassium channels are involved, Tl2SO4 needs to be adjusted. + and K + The concentration of Tl + The concentration is about 0.5mM~5mM, K+ The concentration is about 5mM to 30mM. 2) Each 1mM solution of K2SO4 and Tl2SO4 contains 2mM K + and Tl + The final concentration must be calculated based on the ion concentration. 3) TlCl is prone to precipitation and requires a chloride-free buffer.

[0180] (3) The next day, experimental test: After the cells reach confluence, remove the cell analysis plate from the incubator. Prepare the experimental buffer and dilute Tl + Dye (Molecular Devices #R8222). Discard the culture medium and add 25 μL Tl + After 1 hour of incubation, the cell assay plate, composite plate, and FLIPR tip were placed in a FLIPR Tetra (Molecular Devices, USA) for FLIPR detection. After the tip was set up, a baseline reading was performed for 60 seconds, and then 12.5 μL from the composite plate (3×) was added to the cell assay plate while recording data for at least 5 minutes.

[0181] (4) Data processing: Maximum signal was generated by FLIPR software. Data analysis was performed using Excel (2013) and Prism 6.01. Data quality control was S / B>2.00 and Z factor>0.50. IC 50 or EC 50 It was defined as the midline between the lowest and highest plateaus and was calculated using a four-parameter logistic model using Prism 6.01.

[0182] Test Method 2: Pharmacokinetic Evaluation in Mice

[0183] Experimental Methods: The test compound was mixed with 5% DMSO / 60% polyethylene glycol (PEG400) / 35% water, vortexed, and sonicated to obtain a 0.2 mg / mL clear solution. The solution was then filtered through a microporous filter and used for later use. Male CD-1 mice, weighing 25 to 35 grams, were administered intravenously with the candidate compound solution at a dose of 1 mg / kg. The mice were fasted and then orally administered with the candidate compound solution at a dose of 5 mg / kg. After dosing, 25 μL of blood was collected from the animals at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours and placed into commercially available tubes pre-coagulated with EDTA-K2. The tubes were centrifuged for 10 minutes to separate plasma and store at -60°C. The target compound content in the plasma samples was determined by LC / MS / MS, and pharmacokinetic parameters were calculated using Phoenix WinNonlin software (Pharsight, USA).

[0184] Definition of each parameter: IV: intravenous injection; PO: oral administration; C0: required concentration immediately after intravenous injection; Cmax: maximum blood drug concentration after administration; T max : time required to reach peak drug concentration after administration; T 1 / 2 : The time required for the blood drug concentration to drop by half; Vdss: Apparent distribution volume, which refers to the proportional constant between the amount of drug in the body and the blood drug concentration when the drug reaches dynamic equilibrium in the body. Cl: Clearance, which refers to the apparent distribution volume of the drug cleared from the body per unit time; T last : The time of the last detection point; AUC 0-last : The area under the drug-time curve refers to the area enclosed by the blood drug concentration curve relative to the time axis; F: A measure of the speed and extent to which the drug is absorbed into the blood circulation, and is an important indicator for evaluating the degree of drug absorption.

[0185] Technical Effects

[0186] The compound of the present invention has good binding to Kv7.2 protein and human Kv7.3 protein with highly homologous binding sites, has a significant agonist effect on hKCNQ2 / 3 potassium ion channels, and has good pharmacokinetic properties, and can exert good pharmacodynamic effects.

[0187] Definition and Description

[0188] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0189] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0190] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, which are prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases, having specific substituents. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or a suitable inert solvent. Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts.

[0191] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0192] A "pharmaceutical composition" refers to a composition containing one or more compounds described herein, their isomers, or pharmaceutically acceptable salts thereof, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0193] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0194] The pharmaceutical composition of the present invention can be prepared by combining the compound of the present invention with suitable pharmaceutically acceptable excipients.

[0195] The pharmaceutical composition of the present invention can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, sugar-coated pill making methods, grinding methods, emulsification methods, freeze-drying methods, etc.

[0196] The term "treatment" means administering the compounds or formulations of the present invention to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0197] (i) inhibiting a disease or disease state, i.e., arresting its development;

[0198] (ii) ameliorating the disease or condition, i.e., causing regression of the disease or condition.

[0199] The term "prevention" means administering a compound or formulation of the invention to prevent a disease or one or more symptoms associated with the disease, including preventing the disease or disease state from occurring in a mammal, particularly when such mammal is susceptible to the disease state but has not yet been diagnosed as having the disease state.

[0200] The term "therapeutically effective amount" means an amount of a compound of the present invention that (i) treats or prevents a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder as described herein. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art based on their knowledge and the disclosure herein.

[0201] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0202] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0203] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0204] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0205] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0206] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed bond Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond or straight dashed key

[0207] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of that isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0208] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0209] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0210] Certain compounds of the present invention may exist as atropisomers, which are conformational isomers that occur when rotation about a single bond in a molecule is prevented or greatly slowed due to steric interactions with other parts of the molecule. The compounds disclosed herein include all atropisomers, either pure individual atropisomers, or enriched in one atropisomer, or as nonspecific mixtures of each. If the rotational potential about the single bond is sufficiently high and the interconversion between conformations is sufficiently slow, separation of the isomers may be permitted.

[0211] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125I) or C-14( 14 C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0212] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be any on a chemically feasible basis.

[0213] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0214] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0215] When the number of a substituent is 0, it means that the substituent does not exist, for example, -A-(R)0 means that the structure is actually -A.

[0216] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A.

[0217] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0218] When the linking group is listed without specifying its linking direction, its linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of reading from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0219] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bond is connected, the number of H atoms at the site will decrease accordingly with the number of connected chemical bonds, and become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy lines For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if an H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

[0220] Unless otherwise specified, the number of atoms in a ring is generally defined as the number of members of the ring, for example, a "5-7 membered ring" refers to a "ring" having 5-7 atoms arranged around it.

[0221]

[0046] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0222] Unless otherwise specified, the term “C 1-6 "Alkyl" itself or in combination with other terms is used to refer to a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1-6 Alkyl groups include C 1-2 、C 1-3 、C 1-4 、C 1-5 、C 2-3 、C 2-4 , C3, C4, C5, C6 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), and the like.

[0223] Unless otherwise specified, the term “C 1-4 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms. 1-4 Alkyl groups include C 1-2 、C 1-3 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-4 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), and the like.

[0224] Unless otherwise specified, the term “C 2-4 "Alkyl" by itself or in combination with other terms refers to a straight or branched chain saturated hydrocarbon group consisting of 2 to 4 carbon atoms. 2-4 Alkyl groups include C 2-4 and C 2-3 Alkyl, etc.; it can be monovalent (such as ethyl), divalent (such as ethylene) or polyvalent (such as ethylene). 2-4 Examples of alkyl groups include, but are not limited to, ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), and the like.

[0225] Unless otherwise specified, the term “C 1-4 "Alkoxy" by itself or in combination with other terms refers to an alkyl group containing 1 to 4 carbon atoms, which is attached to the rest of the molecule through an oxygen atom. 1-4 Alkoxy groups include C 1-3 、C 1-2 、C 2-4 , C4 and C3 alkoxy, etc. It can be monovalent, divalent or polyvalent. 1-4 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), and the like.

[0226] Unless otherwise specified, the term “C 1-6 "Alkoxy" refers to an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an oxygen atom. 1-6 Alkoxy groups include C 1-4 、C 1-3 、C1-2 、C 2-6 、C 2-4 , C6, C5, C4 and C3 alkoxy, etc. 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexyloxy, and the like.

[0227] Unless otherwise specified, “C 3-7 "Cycloalkyl" itself or in combination with other terms refers to a saturated cyclic hydrocarbon group consisting of 3 to 7 carbon atoms. It contains a single ring or multiple rings, wherein the multiple rings include spiro rings, cyclic rings and bridged rings. The C 3-7 Cycloalkyl groups include C 3-5 、C 3-6 、C 4-5 、C 4-6 、C 5-7 , C4, C5, C6, C7 cycloalkyl, etc.; which may be monovalent, divalent or polyvalent. 3-7 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wait.

[0228] Unless otherwise specified, “C 4-7 "Cycloalkyl" itself or in combination with other terms refers to a saturated cyclic hydrocarbon group consisting of 4 to 7 carbon atoms. It contains a single ring or multiple rings, wherein the multiple rings include spiro rings, cyclic rings and bridged rings. The C 4-7 Cycloalkyl groups include C 4-5 、C 4-6 , C4, C5, C6, C7 cycloalkyl, etc.; which may be monovalent, divalent or polyvalent. 4-7 Examples of cycloalkyl groups include, but are not limited to, cyclobutyl, cyclopentyl, cyclohexyl, wait.

[0229] Unless otherwise specified, “C 5-8 "Bridged ring cycloalkyl" means a saturated bridged ring hydrocarbon group consisting of 5 to 8 carbon atoms. 5-8 Bridged cycloalkyl groups include C 5-6 、C 5-7 、C 5-8 、C 6-7 、C 6-8 or C 7-8 Bridged ring cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 5-8 Examples of bridged cycloalkyl groups include, but are not limited to, wait.

[0230] Unless otherwise specified, “C5-8 Spirocyclic cycloalkyl" refers to a saturated spirocyclic hydrocarbon group consisting of 5 to 8 carbon atoms. 5-8 Spirocyclic cycloalkyl groups include C 5-6 、C 5-7 、C 5-8 、C 6-7 、C 6-8 or C 7-8 Spirocyclic cycloalkyl, etc.; which may be monovalent, divalent or polyvalent. 5-8 Examples of spirocyclic cycloalkyls include, but are not limited to, wait.

[0231] Unless otherwise specified, the term "4-7 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated cyclic group consisting of 4 to 7 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the carbon atoms are optionally oxo (i.e., C(O)), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). It includes monocyclic or polycyclic rings, wherein polycyclic rings include spirocyclic rings, fused rings, and bridged rings. In addition, with respect to the "4-7 membered heterocycloalkyl", a heteroatom may occupy the position at which the heterocycloalkyl is connected to the rest of the molecule. The 4-7 membered heterocycloalkyl includes 4-5 membered, 4-6 membered, 5-6 membered, 5-7 membered, 4 membered, 5 membered, 6 membered, 7 membered heterocycloalkyl, etc. Examples of 4-7 membered heterocycloalkyl groups include, but are not limited to, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, wait.

[0232] Unless otherwise specified, “C 5-8 "Cycloalkenyl" means a partially unsaturated cyclic hydrocarbon group consisting of 5 to 8 carbon atoms containing at least one carbon-carbon double bond, which includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spirocyclic, fused and bridged rings, and any ring of this system is non-aromatic. The C 5-8 Cycloalkenyl groups include C 5-6 、C 5-7 、C 6-7 、C 6-8 、C 7-8 Cycloalkenyl, etc.; which may be monovalent, divalent or polyvalent. 5-8Examples of cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like.

[0233] Unless otherwise specified, “C 4-7 "Cycloalkenyl" means a partially unsaturated cyclic hydrocarbon group consisting of 4 to 7 carbon atoms containing at least one carbon-carbon double bond, which includes monocyclic and polycyclic ring systems, wherein the polycyclic ring system includes spirocyclic, fused and bridged rings, and any ring of this system is non-aromatic. The C 4-7 Cycloalkenyl groups include C 4-5 、C 5-6 , C4, C5, C6, C7 cycloalkenyl, etc.; which may be monovalent, divalent or polyvalent. 4-7 Examples of cycloalkenyl groups include, but are not limited to, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like.

[0234] Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" are used interchangeably in the present invention. The term "5-6 membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 5 to 6 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p , p is 1 or 2). 5-6 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. Examples of the 5-6 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H- 1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0235] Unless otherwise specified, the term "4-7 membered heterocycloalkenyl" by itself or in combination with other terms refers to a partially unsaturated cyclic group consisting of 4 to 7 ring atoms containing at least one carbon-carbon double bond, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the carbon atoms are optionally oxoed (i.e., C(O)), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). It includes monocyclic or polycyclic rings, wherein the polycyclic ring system includes spirocyclic, fused and bridged rings, and any ring of the system is non-aromatic. In addition, with respect to the "4-7 membered heterocycloalkenyl", a heteroatom may occupy the position where the heterocycloalkenyl group is connected to the rest of the molecule. The 4-7 membered heterocycloalkenyl group includes 4-5 membered, 4-6 membered, 5-6 membered, 5-7 membered, 4 membered, 5 membered, 6 membered and 7 membered heterocycloalkenyl groups. Examples of 4-7 membered heterocycloalkenyl groups include, but are not limited to

[0236] Unless otherwise specified, the terms "9-10 membered bicyclic heteroaromatic ring" and "9-10 membered bicyclic heteroaryl" are used interchangeably. The term "9-10 membered heteroaryl" refers to a cyclic group consisting of 9 or 10 ring atoms with a conjugated π electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. It is a bicyclic ring system in which each ring is aromatic. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). The 9-10 membered heteroaryl group may be attached to the remainder of the molecule via a heteroatom or carbon atom. The 9-10 membered bicyclic heteroaryl group includes 9-membered and 10-membered bicyclic heteroaryl groups. Examples of the 9-10 membered bicyclic heteroaryl group include, but are not limited to, benzimidazolyl, benzopyrazolyl, pyridoimidazolyl, pyridopyrazolyl, and the like.

[0237] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0238] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compounds, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data on the cultured single crystal using a Bruker D8 venture diffractometer, using CuKα radiation as the light source and scanning mode: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0239] Abbreviations used in the present invention: prep-HPLC stands for preparative high performance liquid chromatography; DPPA stands for diphenylphosphoryl azide.

[0240] The solvents used in the present invention can be obtained commercially. Software naming, commercially available compounds use supplier catalog names. BRIEF DESCRIPTION OF THE DRAWINGS

[0241] Figure 1 shows the binding pattern of compound A to KCNQ2;

[0242] Figure 2 shows the binding pattern of compound B and KCNQ2;

[0243] Figure 3 shows the binding pattern of compound C and KCNQ2;

[0244] Figure 4 shows the binding pattern of compound D to KCNQ2;

[0245] Figure 5 shows the binding pattern of compound E and KCNQ2;

[0246] Figure 6 shows the binding pattern of compound F and KCNQ2;

[0247] Figure 7 shows the binding pattern of compound G to KCNQ2. DETAILED DESCRIPTION

[0248] The present invention is described in detail below by way of examples, but is not intended to limit the present invention in any way. While the present invention has been described in detail herein, and specific embodiments thereof have been disclosed, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0249] Calculation Example 1

[0250] By using Maestro( Glide SP in version 2022-1 [1] The co-crystal structure of KCNQ2 and Retigabine in the PDB database PDB:7CR2 was selected as the docking template after energy optimization. [2] The protein preparation wizard module was used to add hydrogen atoms and the OPLS4 force field was used. For ligand preparation, the three-dimensional structure of the molecule was generated using LigPrep and energy minimization was performed.[3] , using the Confgen module to sample small molecule conformations. Using Retigabine in 7CR2 as the center of mass, a 3D image with a side length of A cubic docking grid. During molecular docking, compounds were placed within the protein model, and the interaction types between the protein receptor and ligand were analyzed. Then, based on the calculated docking scores and binding modes, reasonable docking conformations were selected and saved. The binding modes of compounds A–G are shown in Figures 1 to 7.

[0251] [1]Glide, LLC, New York, NY, 2022.

[0252] [2]Maestro, LLC, New York, NY, 2022.

[0253] [3]LigPrep, LLC, New York, NY, 2022.

[0254] Conclusion: The compounds of the present invention have good binding to KCNQ2.

[0255] Example 1

[0256] Step 1: Compound 1-1 (10 g, 42.02 mmol) was added to ethanol (200 mL), followed by 1-2 (9.83 g, 50.42 mmol). The mixture was heated to 85°C under nitrogen for 12 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate = 100 / 0 to 10 / 90, to dichloromethane / methanol = 100 / 0 to 5 / 95, V / V) to obtain compound 1-3. LCMS (ESI): 335.0 [M+H] + .

[0257] Step 2: Compound 1-3 (1.14 g, 3.41 mmol) was added to N,N-dimethylacetamide (20 mL), followed by zinc cyanide (601.01 mg, 5.12 mmol), tetrakistriphenylphosphine palladium (197.16 mg, 170.62 μmol), and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (124.84 mg, 170.62 μmol). The reaction mixture was heated to 130°C under nitrogen for 2 hours. The reaction mixture was cooled to room temperature and filtered under reduced pressure. The filtrate was collected and concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 50 / 50, V / V) to obtain compound 1-4. LCMS (ESI): 234.0 [M+H]+ .

[0258] Step 3: Compound 1-4 (1.5 g, 6.43 mmol) was added to N,N-dimethylformamide (20 mL). After cooling to 0°C in an ice-water bath, N-bromosuccinimide (1.37 g, 7.72 mmol) was added. The reaction mixture was heated to 25°C under nitrogen for 2 hours. Water (40 mL) was added to the reaction mixture, and solid precipitated. The mixture was filtered under reduced pressure and the filter cake was collected to obtain compound 1-5. LCMS (ESI): 312.0 [M+H] + .

[0259] Step 4: Compound 1-5 (1 g, 3.20 mmol) was added to methanol (1.5 mL), tetrahydrofuran (3 mL), and water (3 mL), followed by the addition of lithium hydroxide (153.48 mg, 6.41 mmol). The reaction mixture was reacted at 25°C under nitrogen for 1 hour. After the reaction was complete, hydrochloric acid (10 mL, 2 M) was added dropwise to quench the reaction. Water (100 mL) was then added, and the mixture was extracted with ethyl acetate (100 mL). The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered under reduced pressure, and evaporated to dryness to obtain compound 1-6. LCMS (ESI): 284.1 [M+H] + .

[0260] Step 5: Compound 1-6 (270 mg, 950.57 μmol) was added to tert-butanol (5 mL), followed by N,N-diisopropylethylamine (147.42 mg, 1.14 mmol) and diphenylphosphoryl azide (313.91 mg, 1.14 mmol). The reaction mixture was heated to 85°C under nitrogen for 12 hours. The reaction mixture was concentrated under reduced pressure, and ethyl acetate (100 mL) was added to the residue. The mixture was washed with water (100 mL x 3). The organic phase was collected and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 20 / 80, v / v) to obtain compound 1-7. LCMS (ESI): 356.9 [M+H] + .

[0261] Step 6: Compound 1-7 (50 mg, 140.78 μmol) was added to dichloromethane (1 mL), and trifluoroacetic acid (0.5 mL) was added at 0°C. The reaction mixture was reacted at 25°C under nitrogen for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a crude product. Water (10 mL) and ethyl acetate (20 mL) were added, and the pH was adjusted to alkaline using saturated sodium bicarbonate. The mixture was washed with water (10 mL × 3). The organic phase was collected and then washed with saturated saline solution (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 50 / 50, V / V) to obtain compound 1-8. LCMS (ESI): 254.9 [M+H] + .

[0262] Step 7: Compound 1-8 (35 mg, 137.23 μmol) was added to acetone (4 mL), and N,N-diisopropylethylamine (19.51 mg, 150.95 μmol) and tert-butylacetyl chloride (19.40 mg, 144.09 μmol) were added sequentially at 0°C. The reaction mixture was reacted at 25°C under nitrogen for 2 hours. Water (25 mL) was slowly added to the reaction mixture, and the mixture was extracted with ethyl acetate (25 mL x 2). The combined organic phases were washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 50 / 50, V / V) to obtain compound 1-9. LCMS (ESI): 353.0 [M+H] + .

[0263] Step 8: Compound 1-9 (32 mg, 90.60 μmol) was added to dioxane (5 mL), followed by p-trifluoromethoxyphenylboronic acid (27.99 mg, 135.90 μmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (6.63 mg, 9.06 μmol), potassium carbonate (25.04 mg, 181.21 μmol), and water (0.5 mL). The mixture was stirred at 85°C under nitrogen for 2 hours. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated by Prep-HPLC (column: 2-Phenomenex Gemini C18 75*40mm*3μm; mobile phase: [water (ammonia + ammonium bicarbonate)-acetonitrile]; acetonitrile ratio: 50%-80%) to obtain compound 1. LCMS (ESI): 435.0 [M+H] +. 1H NMR (400MHz, CDCl3) δppm 8.27 (s, 1H) 7.54 (d, J = 8.8Hz, 2H) 7.42 (br d, J = 8.8Hz, 2H) 7.11 (d, J = 8.8Hz, 1H) 2.20 (br s, 2H) 0.99 (s, 9H).

[0264] Example 2

[0265] Compound 1-9 (0.11 g, 311.45 μmol) was added to dioxane (5 mL), followed by p-4-pyridineboronic acid (76.56 mg, 622.90 μmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.99 mg, 31.14 μmol), potassium carbonate (86.09 mg, 622.90 μmol), and water (0.5 mL). The mixture was stirred at 85°C under nitrogen for 2 hours. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated by Prep-HPLC (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (ammonia + ammonium bicarbonate)-acetonitrile]; acetonitrile ratio: 18%-48%) to obtain compound 2, LCMS (ESI): 352.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.84 (d, J = 4.0 Hz, 2H), 8.40 (s, 1H), 7.94 (s, 1H), 7.43 (d, J = 5.6 Hz, 2H), 7.16 (d, J = 8 Hz, 1H), 2.23 (s, 2H), 1.03 (s, 9H).

[0266] Example 3

[0267] Step 1: Compound 1-5 (1.33 g, 4.26 mmol) was added to dioxane (20 mL), followed by 2-cyclopentene-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.65 g, 8.52 mmol), tetrakistriphenylphosphine palladium (246.22 mg, 213.08 μmol), potassium carbonate (1.18 g, 8.52 mmol), and water (2 mL). The reaction mixture was heated to 85°C under nitrogen for 2 hours. Ethyl acetate (200 mL) was added to the reaction mixture, and the mixture was washed with water (200 mL × 3). The organic phase was collected and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 20 / 80, v / v) to obtain compound 3-1. LCMS (ESI): 300.1 [M+H] + .

[0268] Step 2: Compound 3-1 (0.95 g, 3.17 mmol) was added to methanol (1.5 mL), tetrahydrofuran (3 mL), and water (3 mL). Lithium hydroxide (152.03 mg, 6.35 mmol) was added, and the reaction mixture was reacted at 25°C under nitrogen for 1 hour. After the reaction was complete, hydrochloric acid (10 mL, 2 M) was added dropwise to quench the reaction. Water (100 mL) was added, and the mixture was extracted with ethyl acetate (100 mL). The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude compound 3-2. LCMS (ESI): 272.1 [M+H] + .

[0269] Step 3: Compound 3-2 (0.86 g, 3.17 mmol) was added to tert-butanol (20 mL), followed by N,N-diisopropylethylamine (491.71 mg, 3.80 mmol) and diphenylphosphoryl azide (1.05 g, 3.80 mmol). The reaction mixture was heated to 85°C under nitrogen for 6 hours. Ethyl acetate (100 mL) was added to the reaction mixture, and the mixture was washed with water (100 mL × 3). The organic phase was collected and then washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 20 / 80, V / V) to obtain compound 3-3. LCMS (ESI): 343.3 [M+H] + .

[0270] Step 4: Compound 3-3 (0.2 g, 584.17 μmol) was added to dichloromethane (1 mL), and trifluoroacetic acid (0.5 mL) was added at 0°C. The reaction mixture was reacted at 25°C under nitrogen for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the crude compound. Water (10 mL) and ethyl acetate (20 mL) were added, and the pH was adjusted to alkaline with saturated sodium bicarbonate. The mixture was washed with water (10 mL × 3). The organic phase was collected and washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 50 / 50, V / V) to obtain compound 3-4. LCMS (ESI): 243.1 [M+H] + .

[0271] Step 5: Compound 3-4 (100 mg, 412.79 μmol) was added to acetone (5 mL), and N,N-diisopropylethylamine (58.68 mg, 454.07 μmol) and tert-butylacetyl chloride (58.34 mg, 433.43 μmol) were added sequentially at 0°C. The reaction solution was reacted at 25°C under nitrogen for 2 hours. Water (50 mL) was slowly added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 2), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 50 / 50, V / V) to obtain compound 3. LCMS (ESI): 341.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.61(m,1H),8.45(s,1H),7.12(d,J=8.4Hz,1H),6.20(s,1H),2.69-2.67(m,4H),2.31(s,2H),2.14-2.07(m,2H),1.10(s,9H).

[0272] Example 4

[0273] Step 1: Compound 1-5 (1 g, 3.20 mmol) was added to dioxane (10 mL). Potassium cyclopropyltrifluoroborate (948.28 mg, 6.41 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (234.45 mg, 320.42 μmol), and potassium carbonate (885.69 mg, 6.41 mmol) were added sequentially. The reaction mixture was heated to 85°C under nitrogen for 24 hours. Ethyl acetate (200 mL) was added to the reaction mixture, and the mixture was washed with water (200 mL x 3). The organic phase was collected, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude compound. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 20 / 80, v / v) to obtain compound 4-1. LCMS (ESI): 274.1 [M+H] + .

[0274] Step 2: Compound 4-1 (0.69 g, 2.53 mmol) was added to methanol (1.5 mL), tetrahydrofuran (3 mL), and water (3 mL). Lithium hydroxide (120.95 mg, 5.05 mmol) was added, and the reaction mixture was reacted at 25°C under nitrogen for 1 hour. After the reaction was complete, hydrochloric acid (10 mL, 2 M) was added dropwise to quench the reaction. Water (100 mL) was added, and the mixture was extracted with ethyl acetate (100 mL). The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude compound 4-2. LCMS (ESI): 246.1 [M+H] + .

[0275] Step 3: Compound 4-2 (0.4 g, 1.63 mmol) was added to tert-butanol (10 mL), followed by N,N-diisopropylethylamine (252.99 mg, 1.96 mmol) and DPPA (538.71 mg, 1.96 mmol). The reaction mixture was heated to 85°C under nitrogen for 6 hours. Ethyl acetate (100 mL) was added to the reaction mixture, and the mixture was washed with water (100 mL × 3). The organic phase was collected and then washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 20 / 80, V / V) to obtain compound 4-3. LCMS (ESI): 317.1 [M+H] + .

[0276] Step 4: Compound 4-3 (106 mg, 335.09 μmol) was added to dichloromethane (1 mL), and trifluoroacetic acid (0.5 mL) was added at 0°C. The reaction solution was reacted at 25°C under nitrogen protection for 1 hour. The reaction solution was concentrated to obtain a crude compound. Water (10 mL) and ethyl acetate (20 mL) were added, and the pH was adjusted to alkaline using saturated sodium bicarbonate. The mixture was washed with water (10 mL × 3), and the organic phase was collected and then washed with saturated saline solution (100 mL). It was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 50 / 50, V / V) to obtain compound 4-4. LCMS (ESI): 217.1 [M+H] + .

[0277] Step 5: Compound 4-4 (64 mg, 296.00 μmol) was added to acetone (5 mL), and N,N-diisopropylethylamine (42.08 mg, 325.60 μmol) and tert-butylacetyl chloride (43.83 mg, 325.60 μmol) were added sequentially at 0°C. The reaction solution was reacted at 25°C under nitrogen for 2 hours. Water (50 mL) was slowly added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 2). The organic phases were combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 50 / 50, V / V) to obtain compound 4. LCMS (ESI): 315.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.51 (s, 1H), 7.80 (s, 1H), 7.06 (d, J = 8.0Hz, 1H), 2.34 ( s,2H),2.10-2.01(m,1H),1.15(s,9H),1.14-1.07(m,2H),0.63-0.57(m,2H).

[0278] Example 5

[0279] Step 1: Add tetrahydrofuran (10 mL), water (40 mL), compound 5-1 (2 g, 10.47 mmol), and zinc cyanide (1.23 g, 10.47 mmol) to a 100 mL single-necked flask and begin stirring; after nitrogen replacement, methanesulfonic acid (2-di-tert-butylphosphino-2,4,6-triisopropyl-1,1-biphenyl) (2-amino-1,1-biphenyl-2-yl) palladium (II) (831.80 mg, 1.05 mmol) is added, the temperature is raised to 45°C, and the reaction is carried out for 15 hours. The reaction mixture was cooled to room temperature, water (50 mL) was added, and ethyl acetate (30 mL x 2) was added for extraction. The resulting organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (V / V petroleum ether:ethyl acetate = 1:0 to 2:3) to obtain a crude product. Methyl tert-butyl ether (5 mL) was added to the crude product, stirred for 10 minutes, filtered, and the filter cake was dried to obtain 5-2. LCMS (ESI): 138.0 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.19 (s, 1H), 7.43-7.40 (m, 1H), 5.21 (brs, 2H).

[0280] Step 2: Compound 5-3 (331.22 mg, 2.63 mmol) was added to dichloromethane (12 mL) and stirred. Pyridine (941.9 μL, 11.67 mmol), n-butylphosphonic anhydride (2,4,6-tributyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide, CAS: 163755-62-2, 6.31 g, 8.75 mmol, 50% ethyl acetate solution), and compound 5-2 (400 mg, 2.92 mmol) were then added in sequence. The mixture was reacted at room temperature (15°C) for 48 hours. Water (20 mL) and dichloromethane (20 mL) were added to the reaction system, and the mixture was separated. The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (V / V petroleum ether: ethyl acetate = 1:0 to 1:1) to obtain compound 5-4. LCMS (ESI): 246.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.50 (d, J=1.6Hz, 1H), 7.75 (brs, 1H), 7.69 (dd, J=9.2Hz, 1.6Hz, 1H), 2.86 (s, 2H), 2.54 (s, 1H), 1.87 (s, 6H).

[0281] Step 3: Compound 5-4 (20 mg, 81.55 μmol) was added to chlorobenzene (4 mL) and stirred. Then, thionyl chloride (29.61 μL, 407.74 μmol) and pyridine (23.04 μL, 285.42 μmol) were added sequentially. The temperature was raised to 90°C and the reaction was allowed to proceed for 13 hours. The reaction solution was concentrated under reduced pressure at 50°C. The crude product was purified by silica gel column chromatography (gradient elution: petroleum ether:ethyl acetate = 100:0 to 85:15) to obtain compound 5-5. LCMS (ESI): 262.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.38 (s, 1H), 7.05 (dd, J = 9.2Hz, 1.2Hz, 1H), 2.77 (s, 1H), 2.49 (s, 6H).

[0282] Step 4: Compound 5-5 (50 mg, 191.07 μmol) and compound 5-6 (264.08 mg, 2.29 mmol) were added to dioxane (5 mL) and stirred; after nitrogen replacement, cesium carbonate (186.77 mg, 573.22 μmol), potassium iodide (95.15 mg, 573.22 μmol), methanesulfonic acid (2-dicyclohexylphosphino-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl) (2-amino-1,1-biphenyl-2-yl) palladium (II) (34.64 mg, 38.21 μmol) were added in sequence, and the temperature was raised to reflux for 3 hours. After the reaction solution cooled to room temperature, saturated ammonium chloride solution (20 mL) was added to the reaction system, followed by extraction with ethyl acetate (20 mL × 2 times); the resulting organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (V / V petroleum ether: ethyl acetate = 1:0 to 1:1) to obtain a crude product, which was then separated by preparative high-performance liquid chromatography (chromatographic column: WePure Biotech XP tC18 100mm*30mm*10μm; mobile phase: A (water, containing 10mM ammonium bicarbonate) and B (acetonitrile); gradient: B%: 32%-62%) to obtain compound 5. LCMS (ESI): 341.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.42 (s, 1H), 7.33 (brs, 1H), 7.02 (d, J = 9.2Hz, 1H), 2.73 (s, 1H), 2.42 (s, 6H), 2.32 (s, 2H), 1.14 (s, 9H).

[0283] Example 6

[0284] Step 1: Add tetrahydrofuran (25 mL), water (100 mL), compound 6-1 (5 g, 20.75 mmol), and zinc cyanide (4.87 g, 41.49 mmol) to a 250 mL single-necked bottle and start stirring; after nitrogen replacement, methanesulfonic acid (2-di-tert-butylphosphino-2,4,6-triisopropyl-1,1-biphenyl) (2-amino-1,1-biphenyl-2-yl) palladium (II) (1.65 g, 2.07 mmol) is added, the temperature is raised to 45°C, and the reaction is carried out for 15 hours. Water (50 mL) was added, and extraction with ethyl acetate (80 mL x 2) was performed. The resulting organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (V / V petroleum ether:ethyl acetate = 1:0 to 2:3) to obtain a crude product. Methyl tert-butyl ether (10 mL) was then added, stirred for 10 minutes, filtered, and the filter cake was dried to obtain compound 6-2. LCMS (ESI): 188.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.50 (d, J = 1.6Hz, 1H), 7.95 (s, 1H), 5.53 (s, 2H).

[0285] Step 2: Add dichloromethane (10 mL), 5-3 (394.39 mg, 3.13 mmol), and N,N-dimethylformamide (129.52 μL, 1.68 mmol) to a 100 mL three-necked flask and start stirring; after lowering the temperature to 0-5°C, add oxalyl chloride (357.88 μL, 4.09 mmol) dropwise thereto, raise the temperature to 20°C, and react for 1 hour; then add compound 6-2 (450 mg, 2.40 mmol), and then add 4-dimethylaminopyridine (293.80 mg, 2.40 mmol) and triethylamine (2.01 mL, 14.43 mmol) in dichloromethane (3 mL) dropwise, and react at room temperature of 15°C for 18 hours. After adding saturated ammonium chloride solution (30 mL) and dichloromethane (15 mL) to the reaction system, the mixture was separated; the resulting organic phase was washed with saturated ammonium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (V / V petroleum ether:ethyl acetate = 1:0 to 3:2) to obtain compound 6-3. LCMS (ESI): 296.1 [M+H] + .

[0286] Step 3: Add chlorobenzene (36 mL) and compound 6-3 (360 mg, 1.22 mmol) to a 100 mL single-necked flask and begin stirring; then add thionyl chloride (442.78 μL, 6.10 mmol) and pyridine (344.44 μL, 4.27 mmol) in sequence, raise the temperature to 90°C, and react for 15 hours. The reaction solution was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (V / V petroleum ether: ethyl acetate = 1:0 to 17:3) to obtain compound 6-4. LCMS (ESI): 312.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.67(s,1H),7.66(s,1H),2.79(s,1H),2.51(s,6H).

[0287] Step 4: Compound 5-6 (480.37 mg, 4.17 mmol) and compound 6-4 (130 mg, 417.08 μmol) were added to dioxane (8 mL) and stirring was started; after nitrogen replacement, cesium carbonate (407.68 mg, 1.25 mmol), methanesulfonic acid (2-dicyclohexylphosphino-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl) (2-amino-1,1-biphenyl-2-yl) palladium (II) (75.62 mg, 83.42 μmol), potassium iodide (103.85 mg, 625.62 μmol) were added, the temperature was raised to 120 ° C, and the reaction was carried out for 1.5 hours. The reaction mixture was cooled to room temperature, and 20 mL of saturated ammonium chloride solution was added to the reaction system. Ethyl acetate (20 mL x 2) was then added for extraction. The resulting organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (V / V petroleum ether:ethyl acetate = 1:0 to 1:1) and then separated by preparative high-performance liquid chromatography (column: Waters Xbridge BEH C18 100 mm*30 mm*10 μm; mobile phase: A (water containing 10 mM ammonium bicarbonate) and B (acetonitrile); gradient: B%:40%-70%) to obtain compound 6. LCMS (ESI): 391.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 8.73(s,1H),7.80(brs,1H),7.62(s,1H),2.74(s,1H),2.40(s,6H),2.35(s,2H),1.13(s,9H).

[0288] Example 7

[0289] Step 1: Compound 7-A (10 g, 66.60 mmol) was slowly added portionwise to fuming nitric acid (100 mL) and stirred at 0°C for 8 hours. The reaction solution was slowly poured into ice water (500 mL). Solid precipitated, which was filtered, and the filter cake was collected and dried under reduced pressure to obtain compound 7-B. 1 H NMR (CDCl3, 400MHz): δppm 8.45 (d, J = 7.0 Hz, 1H), 7.42 (d, J = 10.0 Hz, 1H), 3.32-3.20 (m, 2H), 2.97-2.79 (m, 2H).

[0290] Step 2: Compound 7-B (1 g, 5.12 mmol) was added to tetrahydrofuran (15 mL), followed by cyclobutylamine (400.89 mg, 5.64 mmol) and N,N-diisopropylethylamine (1.99 g, 15.37 mmol). The mixture was stirred at 25°C for 16 hours. Water (20 mL) was added for dilution, and the mixture was extracted with ethyl acetate (20 mL x 2). The combined organic layers were washed with saturated brine (20 mL), dried over sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 7-C. MS ESI: 246.9 [M+H] + .

[0291] Step 3: Compound 7-C (0.9 g, 3.65 mmol) was added to tetrahydrofuran (15 mL), followed by sodium borohydride (207.40 mg, 5.48 mmol), and stirred at 0°C for 1 hour. Saturated aqueous ammonium chloride (20 mL) was added for dilution, and the aqueous layer was extracted with ethyl acetate (20 mL x 2). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 7-D. MS ESI: 249.0 [M+H] + .

[0292] Step 4: Compound 7-D (0.85 g, 3.42 mmol) was added to trifluoroacetic acid (10 mL), followed by triethylsilane (796.19 mg, 6.85 mmol), and stirred at 25°C for 16 hours. Aqueous sodium bicarbonate (20 mL) was added to dilute the aqueous layer, and the mixture was extracted with ethyl acetate (20 mL x 2). The combined organic layers were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 7-E. MS ESI: 233.1 [M+H] + .

[0293] Step 5: Compound 7-E (0.7 g, 3.01 mmol) was added to ethanol (50 mL) and water (10 mL), followed by reduced iron powder (841.56 mg, 15.07 mmol) and ammonium chloride (806 mg, 15.07 mmol). The mixture was stirred at 90°C for 16 hours. Water (50 mL) was added to dilute the aqueous layer and the mixture was extracted with ethyl acetate (50 mL x 2). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 7-F. MS ESI: 202.9 [M+H] + .

[0294] Step 6: Compound 7-F (0.6 g, 2.97 mmol) was added to ethanol (25 mL), followed by nitrile bromide (628.31 mg, 5.93 mmol), and stirred at 25°C for 16 hours. Aqueous sodium bicarbonate (50 mL) was added for dilution, and the aqueous layer was extracted with ethyl acetate (50 mL x 2). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 7-G. MS ESI: 227.9 [M+H] + .

[0295] Step 7: Compound 7-G (0.15 g, 0.660 mmol) was added to acetone (20 mL), followed by N,N-diisopropylethylamine (252.60 mg, 1.95 mmol) and tert-butylacetyl chloride (131.54 mg, 0.977 mmol), and stirred at 25°C for 16 hours. The reaction mixture was concentrated under reduced pressure, and acetone (10 mL) and aqueous NaOH solution (2N, 20 mL) were added. The mixture was stirred at 25°C for 1 hour and extracted with ethyl acetate (50 mL x 2). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was separated by column chromatography (petroleum ether / ethyl acetate, 1 / 0 to 2 / 1, V / V) to obtain compound 7. MS ESI: 326.1 [M+H] + . 1 H NMR(DMSO-d6,400MHz): δppm 10.26(br s,1H),7.55(br s,1H),7.35(s,1H),4.73(br s,1H),3.00-2.85(m,4H),2.84-2.54(m,2H),2.33(br s,2H),2.26(s,2H),2.09-2.02(m,2H),1.94(q,J=10.2Hz,1H),1.86-1.74(m,1H),1.05(s,9H).

[0296] Example 8

[0297] Step 1: Compound 8-1 (6 g, 26.20 mmol) was added to trifluoroacetic acid (30 mL), followed by triethylsilane (9.14 g, 78.59 mmol). The reaction mixture was incubated at 25°C under nitrogen for 24 hours. Saturated sodium bicarbonate solution was added dropwise to the reaction mixture to adjust the pH to 7-8. The mixture was extracted with ethyl acetate (150 mL x 3), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure to obtain a crude product. The crude product was isolated and purified by column chromatography (petroleum ether / ethyl acetate, 100 / 0, v / v) to obtain compound 8-2.

[0298] Step 2: A 2M solution of lithium diisopropylamine in tetrahydrofuran / n-heptane (22.32 mL) was added to tetrahydrofuran (30 mL). The mixture was cooled to -78°C and compound 8-2 (6.4 g, 29.76 mmol) was added. After stirring for 30 minutes, N,N-dimethylformamide (89.28 mmol, 6.87 mL) was added. The reaction mixture was warmed to 25°C under nitrogen for 30 minutes. Saturated ammonium chloride solution (500 mL) was added dropwise to the reaction mixture. The mixture was extracted with ethyl acetate (100 mL × 3), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure to obtain a crude product. The crude product was isolated and purified by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 5 / 1, v / v) to obtain compound 8-3.

[0299] Step 3: Compound 8-3 (5 g, 20.57 mmol) was added to a mixed solution of water (50 mL) and tert-butanol (50 mL). After cooling to 0°C, sodium chlorite (7.44 g, 82.26 mmol), sodium dihydrogen phosphate (9.87 g, 82.28 mmol) and 2-methyl-2-butene (205.70 mmol, 21.79 mL) were added. The reaction solution was maintained at 0°C under nitrogen protection for 2 hours. The reaction solution was extracted with ethyl acetate (100 mL × 3), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure to obtain compound 8-4. LCMS (ESI): 259.0 [M+H] + .

[0300] Step 4: Compound 8-4 (1.7 g, 6.56 mmol) was added to tert-butanol (17 mL). After cooling to 0°C, N,N-diisopropylethylamine (1.37 mL) and diphenylphosphoryl azide (2.17 g, 7.87 mmol) were added. The reaction solution was heated to 80°C under nitrogen for 2 hours. The reaction solution was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography (petroleum ether / ethyl acetate, 100 / 0 to 20 / 1, V / V) to obtain compound 8-5. LCMS (ESI): 232.0 [M-Boc+H] +. 1H NMR (400MHz, CDCl3) δppm 7.24 (s, 1H), 5.95 (s, 1H), 2.89-2.94 (m, 4H), 2.09-2.16 (m, 2H), 1.51 (s, 9H).

[0301] Step 5: Compound 8-5 (0.4 g, 1.21 mmol) was added to dioxane (5 mL), followed by cyclobutylamine (2.42 mmol, 207.61 μL), (2-dicyclohexylphosphino-2,6-diisopropoxy-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II) methanesulfonate (219.63 mg, 242.28 μmol), 2-dicyclohexylphosphino-2,6-diisopropoxy-1,1-biphenyl (260.10 mg, 484.57 μmol), and potassium tert-butoxide (271.87 mg, 2.42 mmol). The reaction mixture was heated to 100°C under nitrogen for 2 hours. The reaction mixture was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was isolated and purified by column chromatography (petroleum ether / ethyl acetate, 100 / 0-20 / 1, v / v) to obtain compound 8-6. LCMS (ESI): 221.1 [M+H] + .

[0302] Step 6: Compound 8-6 (0.266 g, 1.21 mmol) was added to ethanol (5 mL), followed by cyanogen bromide (2.42 mmol, 177.27 μL). The reaction mixture was maintained at 25°C under nitrogen for 1 hour. The reaction mixture was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography (dichloromethane / methanol, 100 / 0-20 / 1, V / V) to obtain compound 8-7. LCMS (ESI): 246.0 [M+H] + .

[0303] Step 7: Compound 8-7 (55 mg, 179.38 μmol) was added to acetone (5 mL), followed by N,N-diisopropylethylamine (34.37 μL). The reaction solution was cooled to 0°C under nitrogen, and tert-butylacetyl chloride (27.41 μL) was added dropwise. The reaction solution was maintained at 25°C under nitrogen for 1 hour. The reaction solution was filtered under reduced pressure, and the filtrate was collected and dried under reduced pressure. Methanol (5 mL) and 10% aqueous sodium hydroxide solution (2 mL) were added. After stirring for 1 hour, the mixture was dried under reduced pressure and extracted with ethyl acetate (10 mL × 3). The mixture was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and dried under reduced pressure to obtain the crude product. The crude product was separated and purified by column chromatography (petroleum ether / ethyl acetate, 100 / 0-10 / 1, v / v) to obtain compound 8. LCMS (ESI): 344.3 [M+H] + . 1H NMR(400MHz,DMSO-d6)δppm 10.36(s,1H),7.44(s,1H),4.70-4.79ppm(m,1H),3.03(t,J=7.2Hz,2H),2.96(t,J=7.2Hz,2H),2.72-2.85ppm(m,2H) ,2.38-2.40ppm(m,2H),2.29(s,2H),2.11-2.13ppm(m,2H),1.92-1.97ppm(m,1H),1.81-1.86ppm(m,1H),1.066(s,9H)

[0304] Example 9

[0305] Step 1: Compound 9-1 (10 g, 66.60 mmol) was added to fuming nitric acid (100 mL) in batches and stirred at 0°C for 8 hours. The reaction solution was slowly poured into ice water (500 mL), filtered, and the filter cake was collected and dried under reduced pressure to obtain compound 9-2. 1 H NMR (400MHz, CDCl3) δppm 8.44 (d, J = 7.2 Hz, 1H), 7.42 (d, J = 10.4 Hz, 1H), 3.27 (t, J = 6.0 Hz, 2H), 2.81-2.84 (m, 2H).

[0306] Step 2: Compound 9-2 (1 g, 5.12 mmol) was added to tetrahydrofuran (15 mL), followed by 1-methylcyclobutane hydrochloride (972.00 mg, 6.15 mmol) and N,N-diisopropylethylamine (2.68 mL). The reaction mixture was stirred at 25°C for 16 hours. Water (20 mL) was added for dilution, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (20 mL × 2), dried over sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 9-3. LCMS (ESI): 261.1 [M+H] + .

[0307] Step 3: Compound 9-3 (0.5 g, 1.92 mmol) was added to trifluoroacetic acid (5 mL), and triethylsilane (613.63 μL) was added. The reaction solution was kept at 25°C under nitrogen for 24 hours. The reaction solution was cooled to 0°C, and saturated sodium bicarbonate solution was added dropwise to adjust the pH to 7-8. The solution was extracted with ethyl acetate (50 mL × 2), washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure to obtain compound 9-4.

[0308] Step 4: Compound 9-4 (0.8 g, 3.25 mmol) was added to ethanol (40 mL) and water (10 mL). Reduced iron powder (907.01 mg) and ammonium chloride (868.68 mg) were then added. The reaction mixture was heated to 90°C and stirred for 16 hours. Water (200 mL) was added for dilution and extraction was performed with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield compound 9-5.

[0309] Step 5: Compound 9-5 (0.7 g, 3.24 mmol) was added to ethanol (10 mL), followed by nitrile bromide (685.51 mg, 6.47 mmol). The reaction mixture was stirred at 25°C for 12 hours. After dilution with ethyl acetate (50 mL), the mixture was quenched with 10% aqueous sodium bicarbonate solution (50 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 2), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 9-6.

[0310] Step 6: Add tert-Butylacetyl chloride (209.16 mg, 1.55 mmol, 215.85 μL) to acetone (10 mL), followed by N,N-diisopropylethylamine (541.30 μL) and compound 9-6 (0.25 g, 1.04 mmol), and stir at 25°C for 16 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was added to acetonitrile (5 mL), and aqueous sodium hydroxide solution (1 M, 4.57 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. Water (20 mL) was added, and ethyl acetate (20 mL × 3) was added for extraction. The combined organic phases were washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was separated by pre-HPLC (column type: C18 100×40 mm; mobile phase: [H2O(TFA)-ACN]; gradient: 23%-53% ACN, 8 min) to obtain compound 9. LCMS (ESI): 340.1 [M+H] + . 1H NMR (400MHz, CDCl3) δppm 6.93-7.13(m,2H),2.95(t,J=6.8Hz,4H),2.76-2.90(m,2H),2.52(t,J=8.0Hz,2H ),2.31(s,2H),2.06-2.20(m,2H),1.86-2.06(m,3H),1.67(s,3H),1.07ppm(s,9H)

[0311] Example 10

[0312] Step 1: Compound 9-2 (1 g, 5.12 mmol) was added to tetrahydrofuran (20 mL), followed by tert-butylamine (592.32 μL) and N,N-diisopropylethylamine (2.68 mL). The reaction mixture was stirred at 25°C for 16 hours. Water (20 mL) was added for dilution, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with saturated brine (20 mL × 2), dried over sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 10-1.

[0313] Step 2: Compound 10-1 (0.5 g, 2.01 mmol) was added to trifluoroacetic acid (5 mL), followed by triethylsilane (965.00 μL). The reaction mixture was kept at 25°C under nitrogen for 24 hours. The reaction mixture was cooled to 0°C and saturated sodium bicarbonate solution was added dropwise to adjust the pH to 7-8. The mixture was extracted with ethyl acetate (50 mL × 2), saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure to obtain compound 10-2. LCMS (ESI): 179.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.95 (s, 1H), 6.66 (s, 1H), 2.84 (q, J = 7.2Hz, 4H), 2.06-2.10 (m, 2H), 0.98 (s, 1H).

[0314] Step 3: Compound 10-2 (0.1 g, 426.82 μmol) was added to ethanol (30 mL) and water (8 mL). Iron powder (119.19 mg, 2.13 mmol) and ammonium chloride (114.15 mg, 2.13 mmol) were then added. The reaction mixture was heated to 90°C and stirred for 16 hours. Water (200 mL) was added for dilution, and the aqueous phase was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield compound 10-3.

[0315] Step 4: Compound 10-3 (0.3 g, 1.47 mmol) was added to ethanol (5 mL), followed by nitrile bromide (311.06 mg, 2.94 mmol). The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was separated by pre-HPLC (Welch Xtimate C18 150*25mm*5μm; mobile phase: [H2O(NH3.H2O + NH4HCO3)-ACN]; gradient: 33%-63% ACN, 8 min) to obtain compound 10-4.

[0316] Step 5: Add tert-Butylacetyl chloride (16.66 μL) to acetone (10 mL), followed by N,N-diisopropylethylamine (20.89 μL) and compound 10-4 (25 mg, 109.02 μmol), and stir at 25°C for 16 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was added to acetonitrile (5 mL), and aqueous sodium hydroxide solution (1 M, 4.57 mL) was added. The reaction mixture was stirred at 25°C for 2 hours. Water (20 mL) was added, and ethyl acetate (20 mL x 3) was added for extraction. The combined organic phases were washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was separated by pre-HPLC (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [H2O(NH3H2O+NH4HCO3)-ACN]; gradient: 58%-88% ACN, 8 min) to obtain compound 10. LCMS (ESI): 328.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.53(s,1H),7.23-7.05(m,1H),2.92-3.01(q,J=7.2Hz,4H),2.39(s,2H),2.13(t,J=7.6Hz,2H),1.93(s,9H),1.07ppm(s,9H)

[0317] Biological testing

[0318] Experimental Example 1: hKCNQ2 / 3 Thallium Flow Test

[0319] Aim: To determine the in vitro concentration-response relationship of compounds using FLIPR thallium flux analysis in a CHO cell line stably expressing hKCNQ2 / 3.

[0320] Experimental cells and reagents:

[0321] (1) Cell line: CHO cells stably transfected with hKCNQ2 / 3.

[0322] (2) Cell culture medium reagents: including F12 culture medium, fetal bovine serum (FBS) (10%), penicillin-streptomycin (Invitrogen, catalog number 15140-122), and G418 (Invitrogen, catalog number 10131027).

[0323] (3) Cell culture:

[0324] This cell line is passaged three times weekly at a 1:2-1:3 ratio. When cells reach >80% confluency in a T-75 flask, use 0.25% trypsin-EDTA solution for approximately 1 minute and then transfer the cells from the flask for passage. Transfer the cells to another T-75 flask containing complete cell growth medium according to the passage ratio. Note: To maintain logarithmic cell growth, cells should be grown in a subconfluent monolayer. Passage cells every 2-3 days, depending on the cell doubling time.

[0325] Experimental process:

[0326] (1) Day 1, cell preparation: Cells were digested as described above, and cell density and viability were determined using a Cell Countess. The volume of the single-cell suspension was adjusted with complete cell growth medium. hKCNQ2 / 3_CHO cells were then seeded onto PDL-precoated 384-well plates at a density of approximately 20,000 cells / well (30 μL / well). The plates were then incubated overnight in a 37°C, 5% CO2 humidified air incubator.

[0327] (2) The next day, compound preparation: The test compound was diluted to the storage concentration with dimethyl sulfoxide (DMSO) and stored in a refrigerator at -20°C. The compound addition program was set on the liquid workstation ECHO, and the compound was added to the plate using the liquid workstation ECHO. Retigabine (RTG) was used as the positive reference, with the highest concentration being 100 μM, diluted 3-fold, and a total of 9 doses. + and 1 mM Tl + Add the supplemental buffer to the plate.

[0328] Note: 1) K2SO4 and Tl2SO4 are prepared in 1× chloride-free buffer. When different voltage-gated potassium channels are involved, Tl2SO4 needs to be adjusted. + and K + The concentration of Tl + The concentration is about 0.5mM~5mM, K +The concentration is about 5mM to 30mM. 2) Each 1mM solution of K2SO4 and Tl2SO4 contains 2mM K + and Tl + The final concentration should be calculated based on the ion concentration. 3) TlCl is prone to precipitation and a chloride-free buffer should be used.

[0329] (3) The next day, experimental test: After the cells reach confluence, remove the cell analysis plate from the incubator. Prepare the experimental buffer and dilute Tl + Dye (Molecular Devices #R8222). Discard the culture medium and add 25 μL Tl + After 1 hour of incubation, place the cell analysis plate, composite plate, and FLIPR tip on the FLIPR TETRA After setting up the tip, the baseline was read for 60 seconds, and then 12.5 μL from the compound plate (3×) was added to the cell assay plate while recording data for at least 5 minutes.

[0330] (4) Data processing:

[0331] The maximum signal was generated by FLIPR software. Data were analyzed using Excel (2013) and Prism 6.01. Data quality control S / B>2.00, Z factor>0.50. IC 50 or EC 50 It was defined as the midline between the lowest and highest plateaus and was calculated using a four-parameter logistic model using Prism 6.01.

[0332] The experimental results are shown in Table 1.

[0333] Table 1 Experimental results of the agonistic effect of the compounds of the present invention on hKCNQ2 / 3

[0334] Conclusion: The compounds of the present invention have significant agonistic effects on KCNQ2 / 3.

[0335] Experimental Example 2: Evaluating the behavioral pharmacodynamics of the test compound in a maximal electroshock-induced epilepsy model

[0336] Purpose of the experiment:

[0337] This study used male CD-1 mice behaviorally to detect the effects of test compounds on acute epileptic seizure behavior induced by maximal electric shock in mice.

[0338] Experimental Materials:

[0339] Male CD-1 mice (25-35 g) were used as experimental animals and fed with standard feed (Beijing Weitonglihua Experimental Animal Co., Ltd.).

[0340] Experimental process:

[0341] Upon arrival at the animal facility, the animals were acclimated for at least one week. Vital signs of the mice were observed daily, and they were randomly divided into groups based on body weight one day before dosing. Different doses of the test compound were orally administered to each group one hour before electrical stimulation. On the day of testing, the corneas of male CD-1 mice in each group were anesthetized with 2% lidocaine. Seizures were then induced by electrical stimulation of the cornea using a silver bipolar electrode at 18 mA, 60 Hz, a 0.6 ms pulse width, and a stimulation duration of 1 s. The mice were then observed for the latency to seizure and the duration of limb rigidity after electrical stimulation, and the seizure protection rate was calculated for each group.

[0342] The experimental results are shown in the following table:

[0343] Table 2 Pharmacodynamic test results of the compounds of the present invention

[0344] Experimental conclusion: The compound of the present invention has good anticonvulsant activity in the epilepsy model induced by maximum electric shock.

[0345] Experimental Example 3: Pharmacokinetic Evaluation of Compounds

[0346] Purpose of the experiment:

[0347] The purpose of this study is to evaluate the pharmacokinetic behavior of the compound after a single intravenous injection and oral administration, to examine the bioavailability after oral administration, and to provide animal test data for clinical research.

[0348] Experimental Materials:

[0349] CD-1 mice (male, 7-9 weeks old)

[0350] Experimental operation:

[0351] The pharmacokinetic characteristics of the compounds following intravenous and oral administration in rodents were tested using standard protocols. The candidate compounds were prepared as clear solutions and administered to mice as single intravenous and oral injections. The intravenous and oral vehicles were a mixture of 5% DMSO / 60% PEG400 / 35% water. This project used four female CD-1 mice. Two mice were intravenously injected with the drug, and plasma samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. The other two mice were orally gavaged and plasma samples were collected at 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. The samples were stirred at 3,200×g at 4°C for 10 minutes, and the supernatant was separated to obtain plasma samples. Protein was precipitated by adding 20 volumes of methanol solution containing internal standard, stirred at 12,000×g for 15 minutes, and centrifuged at 4°C. 50μL of the supernatant was transferred to a 96-well plate and centrifuged again. The supernatant was sampled and quantitatively analyzed by LC-MS / MS analysis method, and pharmacokinetic parameters such as peak concentration (C max ), clearance (CL), half-life (T 1 / 2 ), tissue distribution (Vdss), area under the drug-time curve (AUC 0-last ), bioavailability (F), etc.

[0352] The experimental results are shown in Table 3:

[0353] Table 3 Pharmacokinetic test results of the compounds of the present invention

[0354] Experimental conclusion: The compounds of the present invention have good pharmacokinetic properties, including good oral bioavailability, oral exposure, half-life and clearance rate.

[0355] Experimental Example 4: Evaluation of Plasma Protein Binding Rate

[0356] Purpose of the experiment:

[0357] The protein binding of the compounds in the plasma of CD-1 mice, Sprague-Dawley rats, beagle dogs, cynomolgus monkeys and humans was determined by equilibrium dialysis.

[0358] Experimental Materials:

[0359] Commercial plasma, double-layer dialysis membrane, and equilibrium dialysis device.

[0360] Experimental procedures:

[0361] Soak the double-layer dialysis membrane in ultrapure water for approximately one hour. Remove the membrane, split it in half, and then soak it in an ethanol:water (20:80, v:v) solution for 20 minutes or store it at 2-8°C. The membrane is valid for one month. Before the experiment begins, rinse the membrane twice with ultrapure water and soak it in ultrapure water for another 20 minutes before use.

[0362] On the day of the experiment, frozen plasma from CD-1 mice, Sprague-Dawley rats, beagle dogs, cynomolgus monkeys, and humans was thawed in running cold tap water. After the plasma was completely thawed, it was centrifuged at 3220 × g for 5 minutes, and suspended solids and precipitates were removed. Plasma pH was measured and used directly if the pH was within the range of 7.4 ± 0.1 or after adjusting the pH with 1% phosphoric acid or 1 M sodium hydroxide to a range of 7.4 ± 0.1.

[0363] Plasma sample preparation: 597 μL of blank plasma from each of the five species was added to 3 μL of the compound or warfarin working solution and mixed thoroughly to obtain plasma samples with a compound concentration of 2 μM and a warfarin concentration of 2 μM (n=1). The organic phase DMSO concentration was 0.5%. The samples were mixed thoroughly before proceeding to the next step.

[0364] Preparation of T0 samples: Pipette 50 μL of compound and warfarin plasma samples into the sample receiving plate (n=3), immediately add 50 μL of blank PBS, then add 500 μL of stop solution to the compound and warfarin T0 samples and store at 2-8°C until further processing with other dialyzed samples.

[0365] Plasma dialysis: 100 μL of compound (2 μM) and warfarin (2 μM) plasma samples were added to the dosing port of each dialysis well (n=3). 100 μL of blank PBS was added to the corresponding receiving port of the dialysis well. The dialysis plate was placed in a 5% CO2 incubator and incubated at 37°C with shaking at approximately 100 rpm for 4 hours.

[0366] After dialysis, 50 μL of dialyzed PBS and dialyzed plasma samples (n = 3) were transferred to a new 96-well plate (sample receiving plate). The corresponding volume of blank plasma or PBS was added to each sample, resulting in a final volume of 100 μL per well, with a plasma-to-PBS volume ratio of 1:1. All samples were subjected to protein precipitation and then analyzed by LC-MS / MS.

[0367] The calculation formulas for the free rate (%Unbound), binding rate (%Bound) and recovery rate (%Recovery) of the compound are as follows:

[0368] %Unbound=100×F / T

[0369] %Bound = 100 - %Unbound

[0370] %Recovery=100×(F+T) / T0

[0371] Where F is the concentration of the compound at the dialysate receiving end; T is the concentration of the compound at the dialysate supply end; and T0 is the concentration of the compound in plasma at time zero.

[0372] The experimental results are shown in Table 4:

[0373] Table 4 Plasma protein binding test results of the compounds of the present invention

[0374] Experimental conclusion: The compound of the present invention has a suitable plasma protein binding rate.

[0375] Experimental Example 5: Permeability Evaluation

[0376] Purpose of the experiment:

[0377] The aim of this study was to evaluate compounds as P-gp substrates in MDR1-MDCK I cells.

[0378] Experimental Materials:

[0379] MDR1-MDCKⅠ cell line.

[0380] Experimental operation:

[0381] Transport experiment: The test article, nadolol, and metoprolol were administered at a concentration of 2.00 μM, and digoxin was administered at a concentration of 10.0 μM. Administration was performed bidirectionally (AB and BA), with two replicates performed for each concentration. The dose solution, receiver solution, and transport buffer were preincubated in a 37.0°C water bath for 30 minutes. The cell layer was rinsed twice with transport buffer. The dose solution and receiver solution were added to the corresponding wells of the cell plate (75.0 and 250 μL per apical and basal wells, respectively). After addition, the cell plate was incubated for 90 minutes in a 37.0°C, 5.0% CO2, and saturated humidity incubator. The initial dose solution was collected as the T0 sample. After 90 minutes of incubation, samples were collected from each well and labeled as AB receiver, BA receiver, AB dose, and BA dose. After sample collection, all samples were mixed with an appropriate volume of transport buffer and stop solution. All samples were vortexed and centrifuged at 3220 × g, 20.0°C for 10 min. An appropriate volume of the supernatant was transferred to a sample analysis plate. After sealing the plate, samples were stored at 2.0–8.0°C if not analyzed immediately. LC-MS / MS analysis was performed.

[0382] Cell Membrane Integrity Test: After the transport experiment, the integrity of the MDR1-MDCK II cell layer was assessed using the Lucifer Yellow Rejection Assay. The remaining solution in the apical and basolateral wells was removed, and 75.0 μL of transport buffer containing 100 μM Lucifer Yellow was added to the apical well and 250 μL of transport buffer was added to the basolateral well. The cell plate was incubated in a cell culture incubator at 37.0°C, 5.0% CO2, and saturated humidity for 30 minutes. A 20.0 μL sample was taken from the apical well and mixed with 60.0 μL of transport buffer. An 80.0 μL sample was taken from the basolateral well and the relative fluorescence unit (RFU) was measured using a microplate reader at 425 / 528 nm (excitation / emission).

[0383] Sample analysis of the compounds and reference compounds, nadolol, metoprolol, and digoxin, was performed using LC-MS / MS. Retention times of the analytes and internal standards, chromatogram acquisition, and chromatogram integration were performed using Analyst software (Sciex, Framingham, MA, USA). Standard curves and quality control samples were not included in the sample analysis, and semiquantitative determination was performed using the ratio of the analyte peak area to the internal standard peak area.

[0384] The apparent permeability coefficient (P app , cm / s), efflux ratio (ER), and recovery rate (Solution Recovery (%) (% Solution Recovery).

[0385] VR is the volume of the receiving solution (0.0750 mL for surface A and 0.250 mL for surface B); Area is the surface area of ​​the cell monolayer (0.143 cm 2 ); Time is the incubation time (5400 s); C0 is the ratio of the initial peak area of ​​the compound or control compound at the dosing site; VD is the volume of the dosing site (0.0750 mL for side A and 0.250 mL for side B); CD and CR are the peak area ratios of the compound or control compound at the dosing site and receiving site, respectively. Lucifer Yellow transmittance (% Lucifer Yellow) was calculated using the following formula:

[0386] RFU Apical and RFU Basolateral are the relative fluorescence intensities of fluorescein at the top and bottom, respectively. Apical With V Basolateral The loading volumes are apical and basolateral (0.0750 and 0.250 mL, respectively).

[0387] The experimental results are shown in Table 5:

[0388] Table 5 Permeability test results of the compounds of the present invention

[0389] Experimental conclusion: The compound of the present invention has good permeability.

[0390] Experimental Example 6: Solubility Evaluation

[0391] Purpose of the experiment:

[0392] The purpose of this experiment was to determine the kinetic solubility of compounds using direct UV assay in 50 mM phosphate buffer (pH 2.0, 6.5, and 7.4) at room temperature for 24 hours.

[0393] Experimental operation:

[0394] Take 10 μL of DMSO stock solution of test compound and control compound and add them to a 96-well plate; add 490 μL of test medium to each 96-well plate, seal the plate, and vortex for 2 minutes. The sample incubation concentration is 200 μM, of which DMSO accounts for 2%. Place the 96-well plate on a shaker at 800 rpm and incubate at room temperature for 24 hours to reach dissolution equilibrium. After incubation, place the sample in a centrifuge and centrifuge at 4000 rpm for 10 minutes at 25°C. Transfer the supernatant of the centrifuged sample to a filter plate, centrifuge at 4000 rpm for 5 minutes, and filter to obtain a filtrate. The concentration of the filtrate is analyzed using LC-UV linear quantitative analysis (sample dilution is performed as needed).

[0395] Experimental data were acquired and processed using Empower software. Linear regression analysis was performed by plotting peak areas against their concentrations in μM.

[0396] The experimental results are shown in Table 6:

[0397] Table 6 Solubility test results of the compounds of the present invention

[0398] Experimental conclusion: The compound of the present invention has suitable solubility.

[0399] Experimental Example 7: Evaluation of liver microsome stability

[0400] Purpose of the experiment:

[0401] The purpose of this study was to evaluate the first-phase metabolic stability of the test article in CD-1 mice, Sprague-Dawley (SD) rats, beagle dogs, cynomolgus monkeys, and human liver microsomes.

[0402] Experimental Materials:

[0403] Animal and human liver microsomes used in this test system were purchased from Xenotech, Corning, or other qualified suppliers.

[0404] Experimental operation:

[0405] Preparation of buffer: Dissolve 73.21 g of potassium phosphate dibasic trihydrate and 10.78 g of potassium dihydrogen phosphate in 4000 mL of ultrapure water. Adjust the pH of the solution to 7.40 ± 0.10 with 10.0% phosphoric acid or 1.00 M potassium hydroxide, for a final concentration of 100 mM.

[0406] Preparation of working solutions: Prepare the test sample powder in DMSO to a 10.0 mM stock solution, which is then diluted with an appropriate organic solvent. Prepare the control compounds, testosterone, diclofenac, and propafenone, in DMSO to a 10.0 mM stock solution, which is then diluted with an appropriate organic solvent.

[0407] Preparation of liver microsomal solution: Dilute each microsome species to a 0.56 mg / mL working solution in 100 mM potassium phosphate buffer. The final concentration of microsomes in the reaction system is 0.500 mg / mL. Detailed preparation protocols are described in the experimental protocol.

[0408] To prepare the reduced nicotinamide adenine dinucleotide phosphate (NADPH) solution, dissolve an appropriate amount of reduced nicotinamide adenine dinucleotide phosphate in magnesium chloride solution and vortex to mix. The final concentrations in the reaction system are 1.00 mM NADPH and 1.00 mM magnesium chloride. The detailed preparation protocol will be described in the experimental notes.

[0409] Preparation of Stop Solution: Prepare the stop solution in acetonitrile containing internal standards (tolbutamide and labetalol). Store the prepared stop solution in a refrigerator at 2.0-8.0°C. The detailed preparation protocol will be described in the experimental log.

[0410] Incubation Procedure: Prepare two 96-well plates, designated T60 and NCF60. Add 445 μL of microsomal working solution (liver microsomal protein concentration 0.56 mg / mL) to each of the T60 and NCF60 plates. Preincubate the plates at 37°C for approximately 10 minutes. After preincubation, add 5 μL of test or control compound working solution to each of the T60 and NCF60 plates and mix thoroughly. Initiate the reaction by adding 50 μL of potassium phosphate buffer to each well of the NCF60 plate. Add 180 μL of stop solution (250 nM tolbutamide and 250 nM labetalol in acetonitrile) and 6 μL of NADPH working solution to the T0 stop plate. Remove 54 μL of sample from the T60 plate and transfer it to the T0 stop plate (T0 sample generation). The reaction was initiated by adding 44 μL of NADPH working solution to each well of the T60 plate. To the blank plate, only 54 μL of microsome working solution, 6 μL of NADPH working solution, and 180 μL of stop solution were added. Therefore, in the test or control compound samples, the final reaction concentrations of the compound, testosterone, diclofenac, and propafenone were 1 μM, the concentration of liver microsomes was 0.5 mg / mL, and the final concentrations of DMSO and acetonitrile in the reaction system were 0.01% (v / v) and 0.99% (v / v), respectively. After incubation for the appropriate time (e.g., 5, 15, 30, 45, and 60 minutes), 180 μL of stop solution (250 nM tolbutamide and 250 nM labetalol in acetonitrile) was added to each well of the stop plate. The reaction was then terminated by removing 60 μL of the sample from the T60 or NCF60 plate. All sample plates were shaken and centrifuged at 3220 × g for 20 min, and then 80 μL of supernatant from each well was diluted into 240 μL of pure water for liquid chromatography tandem mass spectrometry analysis.

[0411] Data Analysis:

[0412] Sample analysis of the compounds in this study was performed using liquid chromatography-tandem mass spectrometry (LC-MS / MS), without a standard curve or quality control samples. Semiquantitative determination was performed using the ratio of the analyte peak area to the internal standard peak area. Retention times of the analytes and internal standards, chromatogram acquisition, and chromatogram integration were performed using Analyst software (Sciex, Framingham, Massachusetts, USA).

[0413] The CV of the internal standard peak area in each matrix should be within 20% for each analytical run.

[0414] The peak height of the highest concentration sample should not exceed 2.00×106 cps (using API 4000 or API 5500) or 4.00×107 cps (using API 6500 or API 6500 Plus). The peak height of the internal standard should not exceed 2.00×106 cps (using API 4000 or API 5500) or 4.00×107 cps (using API 6500 or API 6500 Plus).

[0415] The in vitro elimination rate constants ke of the compounds and control compounds were obtained by converting the ratio of the peak areas of the compound to the internal standard into the residual rate using the following formula:

[0416] when

[0417] The in vitro hepatic microsomal intrinsic clearance (CLint(mic)) and hepatic intrinsic clearance (CLint(liver)) were calculated by ke.

[0418] CLint(mic) = 0.693 / T1 / 2 / microsomal protein content (microsomal concentration during incubation mg / mL)

[0419] CLint(liver) = CLint(mic) × amount of microsomal protein in the liver (mg / g) × liver weight to body weight ratio

[0420] Based on the well-stirred model, the in vivo hepatic clearance (CL(liver)) was estimated as follows:

[0421] CL(liver)=(CLint(liver)×fu×Qh) / (CLint(liver)×fu+Qh)

[0422] By default, fu (free fraction in blood) is equal to 1.

[0423] The experimental results are shown in Table 7:

[0424] Table 7 Liver microsomal stability test results of the compounds of the present invention

[0425] Experimental conclusion: The compound of the present invention has good liver microsome stability.

[0426] Experimental Example 8: Evaluation of drug-drug interactions

[0427] Purpose of the experiment:

[0428] The inhibitory effects of the compounds on the activities of human liver microsomal cytochrome P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A) were evaluated using a cocktail method using specific probe substrates of CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A.

[0429] Experimental Materials:

[0430] Human liver microsomes, probe substrate, positive control inhibitor, and reduced nicotinamide adenine dinucleotide phosphate.

[0431] Experimental procedures:

[0432] Add 20 μL of substrate solution to the corresponding wells of the reaction plate and add 20 μL of potassium phosphate buffer to the blank wells.

[0433] Add 2 μL of test compound and positive control working solution to the corresponding wells, and add 2 μL of solvent to the wells without inhibitor and blank wells.

[0434] Prepare human liver microsome working solution and add 158 μL of human liver microsome working solution to each well of the reaction plate.

[0435] Preheat the reaction plate at 37.0°C for 10 minutes.

[0436] Add 20 μL of NADPH cofactor working solution to the reaction plate to start the reaction.

[0437] After mixing and incubating in a 37.0°C water bath for 10 minutes, 400 μL of stop solution was added to the reaction plate to terminate the reaction.

[0438] The sample plate was placed in a centrifuge and centrifuged at 4000 rpm for 20 minutes.

[0439] Remove 200 μL of the supernatant and add 100 μL of appropriate pure water diluent, and shake the plate until mixed evenly.

[0440] Liquid chromatography-tandem mass spectrometry (LC-MS / MS) was used for analysis.

[0441] Data Analysis:

[0442] The IC of the test sample was calculated by using the three-parameter or four-parameter nonlinear regression analysis in XL fit software with the residual activity percentage as the ordinate and the test sample concentration as the abscissa. 50 When the IC value obtained by XL fit software is 50 Greater than the highest dose concentration (50.0 μM) or unable to fit the IC 50 When IC 50 Values ​​are marked as ">50.0 μM".

[0443] Three-parameter equation:

[0444] Four-parameter equation:

[0445] max: maximum enzyme activity; min: minimum enzyme activity; x: concentration of test article or positive control inhibitor; y: enzyme activity at the corresponding concentration; hillslope: slope; IC 50 : Half inhibition concentration; when the minimum enzyme activity is within ±10%, a four-parameter equation is used, otherwise a three-parameter equation is used. The experimental results are shown in Table 8:

[0446] Table 8 Drug-drug interaction test results of the compounds of the present invention

[0447] Experimental conclusion: The compounds of the present invention have lower CYP inhibition and low risk of drug-drug interaction.

[0448] Experimental Example 9: Fully Automated Patch Clamp (Qpatch) Test of hERG Potassium Channel Effects

[0449] Experimental methods:

[0450] CHO-hERG cells were cultured at 175 cm 2 When the cell density in the culture flask reaches 60-80%, remove the culture medium, wash once with 7 mL PBS (phosphate buffered saline), and then add 3 mL cell dissociation reagent for digestion. After digestion is complete, add 7 mL culture medium to neutralize, then centrifuge, aspirate the supernatant, and add 5 mL culture medium to resuspend to ensure that the cell density is 2-5×10 6 / mL.

[0451] Compound stock solutions were diluted with DMSO. 10 μL of the stock solution was added to 20 μL of DMSO solution and serially diluted 3-fold to six DMSO concentrations. 4 μL of each of the six DMSO concentrations was added to 396 μL of extracellular fluid and diluted 100-fold to six intermediate concentrations. 80 μL of each of the six intermediate concentrations was then added to 320 μL of extracellular fluid and diluted 5-fold to the desired final concentration. The highest concentration tested was 40.00 μM, followed by six concentrations of 40.00, 13.33, 4.44, 1.48, 0.49, and 0.16 μM. The DMSO content in the final test concentration did not exceed 0.2%, as this concentration has no effect on hERG potassium channels. Compound preparation was performed using the Bravo instrument throughout the entire dilution process.

[0452] The Qpatch instrument automatically performed the electrophysiological recording process, including single-cell high-impedance sealing and whole-cell pattern formation. After acquiring the whole-cell recording mode, the cell was clamped at -80 mV. A 50-millisecond pre-voltage of -50 mV was applied before a 5-second depolarizing stimulus of +40 mV. The cell then repolarized to -50 mV for 5 seconds before returning to -80 mV. This voltage stimulus was applied every 15 seconds. After recording for 2 minutes, extracellular solution was added for 5 minutes. Drug administration then began. Compound concentrations were administered for 2.5 minutes at each test concentration, starting with the lowest tested concentration. At least three cells (n ≥ 3) were tested for each concentration.

[0453] Data Analysis:

[0454] The experimental data were analyzed by GraphPad Prism 5.0 software, and the results are shown in Table 9.

[0455] Table 9 IC values ​​of the compounds of the present invention for hERG potassium channel 50 Value test results

[0456] Experimental conclusion: The compound of the present invention has no obvious inhibitory effect on hERG potassium channels and has a low risk of cardiotoxicity.

Claims

1. A compound represented by formula (I) or (V) or a pharmaceutically acceptable salt thereof, in, Ring A is selected from 9-10 membered bicyclic heteroaryl; Ring B is fused with ring A, and ring B is selected from C 4-7 Cycloalkyl, C 4-7 Cycloalkenyl, 4-7 membered heterocycloalkyl and 4-7 membered heterocycloalkenyl; Each R1 is independently selected from H, F, Cl, Br, I, OH, NH2, CN, CF3, C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl, the C 2-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R a replace; R2 is selected from cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 Cycloalkenyl, the cyclobutyl, C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, phenyl, 5-6 membered heteroaryl, 4-7 membered heterocycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b replace; R3 is selected from H and -C(O)C 1-4 alkyl; R4 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, -CH2-C 3-7 Cycloalkyl, -CH2-4-7 membered heterocycloalkyl, each independently optionally substituted by 1, 2, 3, 4 or 5 R c replace; Each R5 is independently selected from H, F, Cl, Br, I, =O, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens; Each R6 is independently selected from H, F, Cl, Br, I, CN, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, 3-7 membered heterocycloalkyl and phenyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, 4-7 membered heterocycloalkyl and phenyl are each independently optionally substituted with 1, 2 or 3 R 6a replace; Each R a and R c Each independently selected from H, F, Cl, Br, I, OH, NH2, CN, CH3 and CF3; Each R b Each independently selected from H, F, Cl, Br, I, =O, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 F; Each R 6a are independently selected from H, F, Cl, Br, I, OH, NH2, CN, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2, 3, 4 or 5 F; m is selected from 1, 2, 3 and 4; p and q are independently selected from 1, 2 and 3; The condition is, 1) When R2 is selected from phenyl and 5-6 membered heteroaryl, the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b When substituted, the structural fragment Selected from or, 2) When R2 is selected from cyclobutyl, the cyclobutyl is optionally substituted by 1, 2 or 3 R b When substituted, m is selected from 2, 3 and 4.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Each R a are independently selected from H, F and OH.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Each R b Each is independently selected from H, F, =O, CH3 and OCF3.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Each R c Each is independently selected from H, F and CH3.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Each R1 is independently selected from H, F, Cl, OH, NH2, CN, CF3, CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl, and the CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl are independently selected from 1, 2 or 3 R a or, each R1 is independently selected from H, F, CN, CF3, 6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R2 is selected from cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridinyl, oxetanyl, azetidinyl, piperidinyl, piperazinyl, cyclopentenyl and cyclohexenyl, wherein the cyclobutyl, spiro[3.3]heptyl, bicyclo[1.1.1]pentyl, phenyl, pyrazolyl, thiazolyl, pyridinyl, oxetanyl, azetidinyl, piperidinyl, piperazinyl, cyclopentenyl and cyclohexenyl are each independently optionally substituted by 1, 2 or 3 R b Substituted; or, R2 is selected from Alternatively, R2 is 7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R4 is selected from CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, CH2C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, The CH2CH3, CH2CH2CH3, CH(CH3)2, C(CH3)3, CH2C(CH3)3, OCH3, OCH2CH3, OCH2CH2CH3, OCH(CH3)2, are independently optionally substituted by 1, 2, 3, 4 or 5 R c Substituted; or, R4 is selected from 8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Structural unit Selected from 9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Structural unit Selected from wherein R5, R6 and p are as defined in claim 1.

10. The compound according to claim 9 or a pharmaceutically acceptable salt thereof, wherein the structural unit Selected from 11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from: in, Each R1 is independently selected from H, F, CN, CF3, R2 is selected from C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, 4-7 membered heterocycloalkyl and C 5-8 Cycloalkenyl, the C 5-8 Bridged ring cycloalkyl, C 5-8 Spirocyclic cycloalkyl, 4-7 membered heterocycloalkyl and C 5-8 The cycloalkenyl group is optionally substituted by 1, 2 or 3 R b replace; R4 is selected from 12. The following compound or a pharmaceutically acceptable salt thereof:

13. A pharmaceutical composition comprising a therapeutically or prophylactically effective amount of the compound according to any one of claims 1 to 12, its stereoisomers and pharmaceutically acceptable salts thereof; further, the pharmaceutical composition also comprises a pharmaceutically acceptable excipient.

14. Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 13, in the preparation of a drug for treating a disease associated with a Kv7 potassium channel opener.

15. The use according to claim 14, wherein: The Kv7 potassium channel opener-related disease is selected from epilepsy, inflammatory pain, neuropathic pain, migraine, depression, anxiety disorder, stroke, Alzheimer's disease, neurodegenerative diseases, neuronal hyperexcitability, complications caused by cocaine abuse, nicotine withdrawal syndrome, alcohol withdrawal syndrome or tinnitus.

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