Ring-fused compound as well as pharmaceutical composition and application thereof

Abstract

The invention relates to a fused ring compound as well as a pharmaceutical composition and application thereof. Specifically disclosed is a compound represented by formula I or a pharmaceutically acceptable salt thereof. The compound provided by the invention has one or more of the following advantages: (1) the structure is novel; (2) the compound has a good retarding effect (inhibiting effect) on the activity of a NaV1.8 channel; and (3) the compound has good selectivity to other subtypes of Nav and is high in safety. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a paracyclic compound, a pharmaceutical composition and application thereof. Background Art

[0002] Pain originates from nociceptors in the peripheral nervous system. This is a free nerve ending widely distributed in the skin, muscles, joints and visceral tissues throughout the body. It can convert the thermal, mechanical or chemical stimulation it senses into nerve impulses (action potentials) and transmit them to the cell body located in the dorsal root ganglion (DRG) via afferent nerve fibers, and finally to higher nerve centers, causing pain. The generation and conduction of action potentials in neurons depends on voltage-gated sodium channels (VGSC / Nav) on the cell membrane. Nav is the key medium for transmembrane information transmission along neurons.

[0003] According to statistics, about one-fifth of the world's population suffers from moderate to severe chronic pain. The global analgesic market was approximately US$36 billion in 2018 and is expected to reach US$56 billion in 2023. Among them, acute moderate to severe pain will grow steadily at a compound annual growth rate of 2.5% in the future, and the chronic pain market will grow at a compound annual growth rate of about 18%. Chronic pain is the main driving force for the continued growth of the global pain market in the next decade.

[0004] The secondary structure of voltage-gated sodium channels (VGSC / Nav) contains four domains (DI-DIV) that form a pore, each of which has six transmembrane α-helices (S1-S6), of which the S4 segment is a voltage sensor containing positively charged ions; the intracellular loop between DIII and DIV is considered to be a fast inactivation gate; Nav exists in three different states, and the closed state of inactivation has different kinetic performances of fast inactivation (within milliseconds) or slow inactivation (seconds). When the cell membrane is depolarized, the sodium ion channel is activated, the channel opens, causing sodium ions to flow in, further depolarizing the cell membrane, and leading to the generation of action potentials. Therefore, inhibiting abnormal sodium ion channel activity helps to treat and relieve pain, and Nav is a potential peripheral target for the treatment of pain.

[0005] There are 9 main subtypes of human Nav, namely Nav1.1-Nav1.9. According to whether it can be effectively inhibited by nanomolar tetrodotoxin (TTX), Nav is divided into TTX-sensitive (TTX-S) and TTX-insensitive (TTX-R), and the tissue expression of different subtypes is extremely different. Nav1.1, Nav1.2, Nav1.3 Nav1.4, Nav1.6 and Nav1.7 are TTX-S types, among which Nav1.1, Nav1.2 and Nav1.3 are expressed in large quantities in the central nervous system (CNS), Nav1.4 is present in large quantities in skeletal muscle, and Nav1.6 and Nav1.7 are mainly present in large quantities in the central nervous system. Nav1.5, Nav1.8 and Nav1.9 are of TTX-R type, among which Nav1.5 is mainly present in cardiomyocytes, and Nav1.8 and Nav1.9 are present in the dorsal root ganglia of the peripheral nervous system (PNS-DRG).

[0006] Studies have shown that changes in ion channels are the molecular basis for peripheral sensitization, central sensitization, and disinhibition after inflammation or neuropathological damage, and are also an important molecular mechanism for the occurrence of pain. Currently, Nav inhibitors have been proven to be effective. For example, the local anesthetic lidocaine relieves pain by inhibiting Nav. Non-selective Nav inhibitors such as lamotrigine, lacosamide, and mexiletine have been successfully used to treat chronic pain. However, the Nav inhibitors currently used in clinical practice lack subtype selectivity and can inhibit sodium ion channels expressed in the heart and central nervous system. The treatment window is narrow and the scope of application is limited. Nav1.8 is an important ion channel involved in chronic pain, atrial fibrillation, and Budd-Chiari syndrome. Highly selective Nav1.8 inhibitors are ideal targets for pain treatment, and when producing analgesic effects, the side effects are relatively small.

[0007] In the model of neuropathic pain, nerve damage increases the expression level of Nav1.8 in axons and neuronal cell bodies. The use of Nav1.8 antisense oligonucleotides can significantly relieve pain while reducing Nav1.8 expression. Nav1.8 knockout mice do not show normal visceral inflammatory pain. When the human Nav1.8 gene produces a gain-of-function mutation, it will cause peripheral neuropathy. Based on a series of animal experiments and human genetic evidence, selective inhibition of Nav1.8 has the potential to become a new type of analgesic therapy, which can be used to treat various types of pain such as inflammatory pain, neuralgia, postoperative pain and cancer pain.

[0008] Highly selective Nav inhibitors are one of the key research and development directions for voltage-gated sodium channels. Nav1.8 is mainly distributed in the peripheral nervous system and is confined to neurons that sense pain. It is a highly selective target for pain treatment, so selectively inhibiting Nav1.8 has good prospects for reducing potential toxic side effects. Nav inhibitors used in clinical practice lack subtype selectivity and can inhibit sodium ion channels expressed in the heart and central nervous system. Therefore, the therapeutic window is narrow and the scope of application is limited. Therefore, it is necessary to develop Nav1.8 inhibitors with higher activity, better selectivity, better pharmacokinetic properties, and fewer side effects to meet clinical needs. Summary of the invention

[0009] The technical problem to be solved by the present invention is to overcome the disadvantage of insufficient inhibition of Nav1.8 channels in the prior art. To this end, the present invention provides a paracyclic compound, a pharmaceutical composition thereof and its application. The compound of the present invention has one or more of the following advantages: (1) novel structure; (2) good blocking effect (inhibition effect) on NaV1.8 channel activity; (3) good selectivity for other subtypes of Nav (such as Nav1.1, Nav1.2, Nav1.3 Nav1.4, Nav1.5, Nav1.6, Nav1.7, Nav1.9) and high safety.

[0010] The present invention solves the technical problem of the present invention through the following technical solutions:

[0011] The present invention provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof,

[0012] ;

[0013] Among them, R 1 , R 2 and R 3 are independently halogen, C 1 -C 6 Alkyl, C 3 -C 6 Cycloalkyl, C 1 -C 6 Alkoxy, -OR 1-1 , by one or more R 1-2 Substituted C 1 -C 6 Alkyl or one or more R 1-3 Substituted C 1 -C 6 Alkoxy;

[0014] R 1-1 C 3 -C 6Cycloalkyl or 3-8 membered heterocycloalkyl; in the 3-8 membered heterocycloalkyl, the heteroatoms are selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0015] Each R 1-2 are each independently a halogen;

[0016] Each R 1-3 Each independently is C 1 -C 6 Alkoxy;

[0017] Ring A is C 6 -C 10 Aryl or 5-10 membered heteroaryl; in the 5-10 membered heteroaryl, the heteroatoms are selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0018] Ring B is a 5-6 membered heterocycloalkyl, a 5-6 membered heterocycloalkenyl or a 5-6 membered heteroaryl; in the 5-6 membered heterocycloalkyl, the 5-6 membered heterocycloalkenyl and the 5-6 membered heteroaryl, the heteroatoms are selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0019] n is 0, 1, 2, 3, 4 or 5 (R 4 The total number of is 0, 1, 2, 3, 4 or 5);

[0020] Each R 4 Each independently is oxo (=O, -O - ), -NH 2 , halogen or -OH;

[0021] R 7 is hydrogen or C 1 -C 6 alkyl;

[0022] R 8 For hydrogen, C 1 -C 6 Alkyl, C 1 -C 6 Alkyl-C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy or -OH.

[0023] In some embodiments, when two R 4 When substituted, the oxo group is =0; that is, together with the attached C, forms C=O.

[0024] In some embodiments, when the N atom is replaced by an R 4When substituted, the oxo is -O - ; That is, together with the connected N, it forms N + -O - .

[0025] In some embodiments, R 1 , R 2 and R 3 wherein the halogen is independently fluorine, chlorine, bromine or iodine, preferably fluorine.

[0026] In some embodiments, R 1 , R 2 and R 3 In the C 1 -C 6 Alkyl and said one or more R 1-2 Substituted C 1 -C 6 C in the alkyl group 1 -C 6 The alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, preferably methyl.

[0027] In some embodiments, R 1 , R 2 and R 3 In the C 3 -C 6 The cycloalkyl groups are each independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, preferably cyclopropyl.

[0028] In some embodiments, R 1 , R 2 and R 3 In the C 1 -C 6 The alkoxy group and the one or more R 1-3 Substituted C 1 -C 6 C in alkoxy 1 -C 6 The alkoxy groups are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy, preferably methoxy or ethoxy.

[0029] In some embodiments, R 1-1 In the C 3 -C 6 The cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, preferably cyclopropyl.

[0030] In some embodiments, R 1-1The heteroatoms in the 3-8 membered heterocycloalkyl are N and / or O; the number of heteroatoms is preferably 1 or 2. The 3-8 membered heterocycloalkyl is preferably a 4-6 membered heterocycloalkyl; more preferably ;For example .

[0031] In some embodiments, R 1-2 In the above, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

[0032] In some embodiments, R 1-3 In the C 1 -C 6 The alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy, preferably methoxy.

[0033] In some embodiments, in Ring A, the C 6 -C 10 Aryl is phenyl or naphthyl; preferably phenyl.

[0034] In some embodiments, in ring A, the heteroatom of the 5-10 membered heteroaryl is N and / or O; the number of heteroatoms is preferably 1 or 2. The 5-10 membered heteroaryl is preferably a 5-6 membered (e.g., 5-membered or 6-membered) heteroaryl; more preferably .

[0035] In some embodiments, in ring B, the heteroatom in the 5-6 membered heterocycloalkyl is N and / or O; the number of heteroatoms is preferably 1 or 2. The 5-6 membered heterocycloalkyl may be a 5 membered heterocycloalkyl or a 6 membered heterocycloalkyl; preferably , , or .

[0036] In some embodiments, in ring B, the heteroatom in the 5-6-membered heterocycloalkenyl group is N and / or O; the number of heteroatoms is preferably 1 or 2. The 5-6-membered heterocycloalkenyl group may be a 5-membered heterocycloalkenyl group or a 6-membered heterocycloalkenyl group; preferably , or .

[0037] In some embodiments, in ring B, the heteroatom in the 5-6 membered heteroaryl group is N and / or O; the number of heteroatoms is preferably 1 or 2. The 5-6 membered heteroaryl group is a 5 membered heteroaryl group or a 6 membered heteroaryl group; preferably or .

[0038] In some embodiments, R 4 In the above, the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine.

[0039] In some embodiments, R 1 C 1 -C 6 Alkoxy.

[0040] In some embodiments, R 2 It is a halogen.

[0041] In some embodiments, R 3 It is a halogen.

[0042] In some embodiments, n is 0, 1, 2, or 3.

[0043] In some embodiments, when R 4 When located on ring A, each R 4 are each independently halogen.

[0044] In some embodiments, when R 4 When located on ring B, each R 4 Each independently is oxo (=O, -O - ), -NH 2 or -OH.

[0045] In some embodiments, the compound represented by Formula I is a compound represented by the following Formula I-1:

[0046] ;

[0047] Among them, R 1 , R 2 , R 3 , n and ring A are as defined above;

[0048] X 1 and X 2 are each independently selected from C or N;

[0049] X 3 , X 4 , X 5 and X 6 Each independently selected from: a chemical bond, O, S, S(O), S(O 2 ), N, NH, CH, NR 4 , CR 4 or C(R 4 ) 2 ; and X 3 , X 4 , X 5 and X 6 At most one of them is a chemical bond at a time;

[0050] Dashed lines represent chemical bonds or none; R4 Oxo (=O, -O - )、-NH 2 , halogen or -OH;

[0051] (When NR 4 When the oxo group is -O - ; That is, NR 4 N + -O - ;

[0052] When C(R 4 ) 2 When the oxo is =O; that is, C(R 4 ) 2 (C=O)

[0053] Among them, R 4 The total number of is 0, 1, 2, 3, 4 or 5.

[0054] In some embodiments, the compound represented by formula I-1 is a compound represented by formula I-2:

[0055] ;

[0056] Among them, ring A, R 4 , n, X 1 , X 2 , X 3 , X 4 , X 5 and X 6 The definition of is as mentioned above.

[0057] In some embodiments, the compound represented by Formula I is a compound represented by the following Formula I-3:

[0058]

[0059] Among them, R 1 , R 2 , R 3 , R 4 , Ring A, Ring B and n are as defined above.

[0060] In some embodiments, the compound represented by formula I-3 is a compound represented by formula I-4:

[0061] ;

[0062] Among them, R 4 , Ring A, Ring B and n are as defined above.

[0063] In some embodiments, R 1It is a methoxy group.

[0064] In some embodiments, R 2 For fluorine.

[0065] In some embodiments, R 3 For fluorine.

[0066] In some embodiments, R 7 It is methyl.

[0067] In some embodiments, R 8 It is methyl.

[0068] In some embodiments, Ring A is or (in," " indicates that ring A is connected to ring B through this bond).

[0069] In some embodiments, Ring B is , , , , , , or (in," " indicates that ring B is connected to ring A through this bond).

[0070] In some embodiments, n is 1, 2, or 3.

[0071] In some embodiments, R 4 Oxo (=O, -O - ), -OH, -NH 2 or F.

[0072] In some embodiments, for , , , , , , , , , or .

[0073] In some embodiments, the compound as shown in Formula I is any of the following compounds:

[0074] .

[0075] The present invention provides a pharmaceutical composition, comprising:

[0076] (1) the compound of formula I or a pharmaceutically acceptable salt thereof, and

[0077] (2) Pharmaceutically acceptable excipients.

[0078] The present invention provides an application of a substance A in preparing a drug for treating a disease; the disease may be pain, a pain-related disease, multiple sclerosis, incontinence or arrhythmia;

[0079] The substance A is the compound shown in formula I or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above.

[0080] In some embodiments, the pain is one or more of acute pain, chronic pain, inflammatory pain, cancer pain, neuropathic pain, musculoskeletal pain, primary pain, intestinal pain, and idiopathic pain.

[0081] The present invention provides an application of the substance A in the preparation of a drug for inhibiting voltage-gated sodium channels; the voltage-gated sodium channels are preferably Na V 1.8.

[0082] The present invention provides an application of the substance A in preparing a voltage-gated sodium channel inhibitor; the voltage-gated sodium channel is preferably Na V 1.8.

[0083] In some embodiments, the voltage-gated sodium channel inhibitor can be used in mammals in vivo; it can also be used in vitro, mainly for experimental purposes, for example: as a standard sample or control sample for comparison, or prepared into a kit according to conventional methods in the art to provide rapid detection of the effect of inhibiting voltage-gated sodium channels.

[0084] The present invention provides an application of the substance A in the preparation of a drug for a disease caused by abnormal activation of a voltage-gated sodium channel; the voltage-gated sodium channel is preferably Na V 1.8; The disease may be pain, a pain-related disease, multiple sclerosis, incontinence or arrhythmia. The pain is preferably as described above.

[0085] Terminology

[0086] In the present invention, the term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. For details, please refer to Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0087] In the present invention, the “ " means that the structural fragment is connected to the rest of the molecule through this bond. For example, It refers to cyclopropyl.

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

[0089] In the present invention, the term "one or more" refers to 1, 2, 3, 4 or 5, such as 1, 2 or 3.

[0090] In the present invention, the term "oxo" refers to "=O" (for example, O and the attached C together form C=O) or "-O - ” (For example, O together with the N to which it is attached forms N + -O - ).

[0091] In the present invention, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0092] In the present invention, the term "alkyl" refers to a group having a specified number of carbon atoms (e.g., C 1 -C 6 ) is a straight or branched, saturated, monovalent hydrocarbon group. The alkyl group includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.

[0093] In the present invention, the term "alkoxy" refers to a group R Y -O-, R Y The same definition as the term "alkyl". Alkoxy includes, but is not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, and the like.

[0094] In the present invention, the term "cycloalkyl" refers to a cycloalkyl group having a specified number of carbon atoms (e.g., 3 -C 6 ), cyclic, saturated, monovalent hydrocarbon group. Cycloalkyl groups include, but are not limited to: , , wait.

[0095] In the present invention, the term "heterocycloalkyl" refers to a cyclic, saturated group with a specified number of ring atoms (e.g., 3-12, 3-10, 4-8, 5, 6 or 7), a specified number of heteroatoms (e.g., 1, 2 or 3), and a specified type of heteroatom (one or more of N, O and S). Heterocycloalkyl is connected to the rest of the molecule through a carbon atom or a heteroatom. Heterocycloalkyl includes, but is not limited to: , , , , , , , wait.

[0096] In the present invention, the term "aryl" refers to a group having a specified number of carbon atoms (e.g., C 6 ~C 10 ) is a cyclic, unsaturated hydrocarbon group, which is a single ring or multiple rings (for example, 2 or 3). When it is a multiple ring, the single rings share two atoms and a bond, and each ring is aromatic. The aryl group is connected to the rest of the molecule through a carbon atom. Aryl groups include, but are not limited to, phenyl and naphthyl.

[0097] The term "heterocycloalkenyl" refers to a cyclic, unsaturated hydrocarbon group having a specified number of ring atoms (e.g., 5-10 members, 5-6 members), a specified number of heteroatoms (e.g., 1, 2, or 3), a specified type of heteroatom (one or more of N, O, and S), and having one or more (e.g., 1, 2, or 3) carbon-carbon sp 2 Double bond, it is a single ring, not aromatic. Heterocycloalkenyl is connected to the rest of the molecule through a carbon atom or a heteroatom. Heterocycloalkenyl includes but is not limited to: , wait.

[0098] The term "heteroaryl" refers to a cyclic, unsaturated group having a specified number of ring atoms (e.g., 5-10 members, 5-6 members), a specified number of heteroatoms (e.g., 1, 2, or 3), a specified type of heteroatom (one or more of N, O, and S), which is a single ring. A heteroaryl group is attached to the rest of the molecule through a carbon atom or a heteroatom. Heteroaryl groups include, but are not limited to: , , wait.

[0099] In the present invention, the term "pharmaceutically acceptable excipients" refers to all substances contained in pharmaceutical preparations other than active pharmaceutical ingredients, which are generally divided into two categories: excipients and additives. For details, please refer to the Pharmacopoeia of the People's Republic of China (2020 Edition) and Handbook of Pharmaceutical Excipients (Paul J Sheskey, Bruno CHancock, Gary P Moss, David J Goldfarb, 2020, 9th Edition).

[0100] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0101] The reagents and raw materials used in the present invention are commercially available.

[0102] The positive and progressive effects of the present invention are that the compounds of the present invention have one or more of the following advantages: (1) novel structure; (2) good blocking effect (inhibitory effect) on NaV1.8 channel activity; (3) good selectivity for other Nav subtypes (such as Nav1.1, Nav1.2, Nav1.3 Nav1.4, Nav1.5, Nav1.6, Nav1.7, Nav1.9) and high safety. DETAILED DESCRIPTION

[0103] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0104] Example 1 BX20-9-007

[0105] Step 1: Synthesis of Intermediate A2

[0106] The synthetic route is shown below:

[0107]

[0108] Method: Compound A1 (18.00 g, 95.76 mmol) was dissolved in toluene (300 mL), and SM1 (13.32 g, 87.07 mmol), Pd (PPh 3 ) 4 (3.19 g, 2.76 mmol), Cu 2O (0.37 g, 2.62 mmol) and K 2 CO 3 (41.19 g, 298.06 mmol), replaced by N 2 The reaction mixture was stirred at 100 °C for 12 h. After the reaction, the mixture was filtered through diatomaceous earth and washed with EA. The filtrate was collected and extracted with saturated brine. The organic phase was dried. After concentration under reduced pressure, it was purified by column chromatography to obtain a white solid compound A2 (7.60 g, 40.4 %).

[0109] Step 2: Synthesis of Intermediate A3

[0110] The synthetic route is shown below:

[0111]

[0112] Method: Compound A2 (7.60 g, 35.16 mmol) was dissolved in THF (80 mL), and lithium hydroxide aqueous solution (3.39 g LiOH·H 2 O was dissolved in 40 mL HO 2 O, 80.87 mmol), the reaction mixture was stirred at 50 ° C for 3 h. After the reaction, DCM was added to dilute the layers, and H 2 The mixture was extracted three times with 4% 4% HCl, the aqueous phases were combined, 1N HCl was added to adjust the pH value to <5, DCM was added for extraction three times, the organic phases were combined, the organic phases were dried, and concentrated under reduced pressure to obtain white solid compound A3 (5.90 g, 83.0 %).

[0113] Step 3: Synthesis of intermediate S2

[0114] The synthetic route is shown below:

[0115]

[0116] Method: Compound S1 (15.50 g, 98.05 mmol) was dissolved in Et 2 O (155 mL) to replace N 2 , cool to about 0 °C, keep the temperature at 0-10 °C and add MeLi.LiBr (209 mL, 313.76 mmol, 1.5 M in Et 2 O), and the temperature was naturally raised to room temperature for 12 h after the reaction was completed. 2 O (100 mL), saturated sodium chloride solution (20 mL) was added to quench, citric acid (25.04 g) was added, and stirred for 0.5 h. Et 2O was extracted three times, the organic phases were combined, dried, and concentrated under reduced pressure to 30 g to obtain crude S2 (in Et 2 O content is about 35%, 30g).

[0117] Step 4: Synthesis of intermediate S3

[0118] The synthetic route is shown below:

[0119]

[0120] Method: Compound A3 (5.90 g, 29.20 mmol) was dissolved in MeCN (210 mL) and stirred until clear. The mixture was cooled in an ice-water bath and CDI (7.00 g, 30.70 mmol) was added to replace N 2 , keep the temperature below 10 °C and stir for 1.5 h, add S2 (in Et 2 O content is about 35%, 15 g), K 2 CO 3 (5.00 g, 36.50 mmol), the reaction mixture was stirred at 35 ° C for 13.5h. After the reaction, water and dilute hydrochloric acid were added to the reaction solution and stirred until clear, extracted with EA, washed with concentrated brine, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to obtain a white solid S3 (7.40 g, 78.7%). MS (ESI, m / z) 323 [M+H] + .

[0121] Step 5: Synthesis of S4

[0122] The synthetic route is shown below:

[0123]

[0124] Method: Compound S3 (9.00 g, 28.0 mmol) was dissolved in anhydrous methanol (800 mL) and anhydrous tetrahydrofuran (160 mL) and stirred until clear. The temperature was lowered to -40 °C and the first batch of NiCl 2 6H 2 O (6.80 g, 28.6 mmol) and NaBH 4 (5.60 g, 148.4 mmol), the second batch of NiCl 2 6H 2 O (6.80 g, 28.6 mmol) and NaBH 4(5.60 g, 148.4 mmol), the reaction is complete after addition. After the reaction is completed, saturated ammonium chloride is slowly added to the reaction solution at -40 °C to quench and stir for 30 min, and the temperature is naturally raised to room temperature, filtered with diatomaceous earth, extracted with DCM DCM, washed with concentrated brine, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to obtain a colorless oily compound S4 (3.57 g, 39.3%). MS (ESI, m / z) 325 [M+H] + .

[0125] The NMR spectrum is:

[0126] 1 H NMR (400 MHz, CDCl 3 ) δ = 6.96-6.83 (m, 2H), 4.47 (d, J = 9.6 Hz,1H), 4.02 (d, J = 2.8 Hz, 1H), 2.92-2.84 (m, 1H), 1.70 (s, 3H), 0.81-0.78 (m,3H).

[0127] Step 6: Synthesis of S5

[0128] The synthetic route is shown below:

[0129]

[0130] Method: Compound S4 (3.57 g, 11.0 mmol) was dissolved in toluene (30 mL) and N 2 , cooled to -30 ° C, DiBAL-H (1.5 M, 7.7 mL, 11.6 mmol) was slowly added dropwise, and the reaction mixture was stirred at -25 ~ -30 ° C for 2 h. After the reaction, the reaction solution was added to saturated ammonium chloride for quenching, filtered through diatomaceous earth, extracted with EA, washed with concentrated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a colorless oily compound S5 (3.53 g, crude product). MS (ESI, m / z) 327 [M+H] + .

[0131] Step 7: Synthesis of S6

[0132] The synthetic route is shown below:

[0133]

[0134] Method: Compound S5 (3.53 g, 10.8 mmol) was dissolved in anhydrous DCM (30 mL) and stirred until clear. Et 3N (2.18 g, 21.6 mmol) was cooled to 0 - 10 °C in an ice - water bath, and AcCl (1.70 g, 21.6 mmol) was slowly added dropwise. After the addition, the mixture was warmed to room temperature and stirred for 1 h. After the reaction was completed, the reaction mixture was quenched with saturated ammonium chloride, extracted with DCM, washed with concentrated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a colorless oily compound S6 (4.36 g, crude product). MS (ESI, m / z) 369 [M + H] + 。

[0135] Step 8: Synthesis of S7

[0136] The synthetic route is shown as follows:

[0137]

[0138] Method: Compound S6 (4.36 g, 11.8 mmol) was dissolved in anhydrous DCM (40 mL) and stirred until clear. N was replaced 2 , and the temperature was cooled to - 78 °C. TMSCN (3.52 g, 35.5 mmol) and BF 3 OEt 2 (5.04 g, 35.5 mmol) were added dropwise in sequence. After the addition, the mixture was stirred at - 60 °C for 13 h and then naturally warmed to room temperature and stirred for 12 h. After the reaction was completed, the reaction mixture was quenched with saturated sodium carbonate solution, filtered through diatomaceous earth, extracted with DCM, washed with concentrated brine, dried over anhydrous sodium sulfate, and concentrated. After purification by column chromatography, a colorless oily compound S7 (2.25 g, 56.9%) was obtained. MS (ESI, m / z) 336 [M + H] + 。

[0139] Step 9: Synthesis of S8

[0140] The synthetic route is shown as follows:

[0141]

[0142] Method: Compound S7 (2.25 g, 6.7 mmol) was dissolved in MeOH (50 mL), and an aqueous KOH solution (2.63 g KOH dissolved in 10 mL H 2 O, 46.9 mmol) was added. The reaction mixture was stirred at 60 °C for 12 h. After the reaction was completed, it was extracted with MTBE. The aqueous phase was adjusted to pH < 5 with hydrochloric acid and then extracted with MTBE. The organic phase was washed with concentrated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a colorless oily compound S8 (2.20 g, crude product). MS (ESI, m / z) 355 [M + H] +

[0143] 1 H NMR (400 MHz, DMSO-d6) δ12.98 (s, 1H), 7.19-7.09 (m, 2H), 4.96 (d,J = 10.4 Hz, 1H), 4.06 (dd, J = 10.4 Hz, 8.0 Hz, 1H), 3.91 (d, J = 2.0 Hz,3H), 2.68-2.60 (m, 1H), 1.51 (s, 3H), 0.68-0.66 (m, 3H)

[0144] Step 10: Synthesis of BX20-9-007

[0145] The synthetic route is shown below:

[0146]

[0147] Method: Compound S8 (0.15 g, 0.42 mmol) was dissolved in DMF (5 mL) and stirred until clear. HATU (0.24 g, 0.64 mmol) and TEA (0.17 g, 1.68 mmol) were added. After stirring for 5 min, Int 7 (0.10 g, 0.64 mmol) was added. The reaction mixture was stirred at 25 °C for 16 h. After the reaction, the reaction solution was added to saturated ammonium chloride to quench, extracted with EA, washed with concentrated brine, dried over anhydrous sodium sulfate, concentrated, and purified by preparative chromatography to obtain a light yellow solid BX20-9-007 (24 mg, 11.8 %). MS (ESI, m / z) 487 [M+H] + .

[0148] 1 H NMR (400 MHz, CD 3 OD) δ 7.84 (d, J = 2.0 Hz, 1H), 7.63-7.60 (m, 1H), 7.26 (d, J = 8.8 Hz, 1H) , 7.19–7.14 (m, 1H), 7.03-6.96 (m, 1H), 5.08 (d, J =10.4 Hz, 1H), 4.35-4.30 (m, 1H), 4.01 (d, J = 2.0 Hz, 3H), 3.23-3.17 (m, 1H), 2.85-2.76 (m, 1H), 1.68 (s, 3H), 0.85-0.81 (m, 3H).

[0149] Example 2 BX20-9-008

[0150] Step 1: Synthesis of Intermediate S8

[0151] Refer to Steps 1 to 9 of Example 1 to synthesize Intermediate S8

[0152] Step 2: Synthesis of Compound BX20-9-008

[0153]

[0154] Method: Dissolve S8 (150 mg, 0.42 mmol) in DCM (5 mL) and DMF (20 μL), slowly add dropwise (COCl) 2 (0.2 mL, 2.13 mmol), react at room temperature for 1 h, concentrate the reaction solution under reduced pressure to obtain the crude product S9 and then dissolve it in DCM (5 mL). Slowly add dropwise the above DCM solution of S9 to the DCM (3 mL) and NMP (3 mL) solution of Int 8 (200 mg, 1.34 mmol), and react at room temperature for 1 h. After the reaction is completed, concentrate the reaction solution under reduced pressure at 40 °C, add EA (30 mL×3) to the residue for extraction, combine the organic phases, wash with H 2 O, then wash with saturated NaCl solution, dry with anhydrous Na 2 SO 4 , purify by preparative chromatography (basic preparation), and obtain a white solid powder after lyophilization (48 mg, yield 23.3%). MS (ESI, m / z) 485.1 [M+H] + .

[0155] 1 H NMR (400MHz, CD 3 OD) δ = 7.93 (d, J = 2.0 Hz, 1H), 7.56 (dd, J = 2.0Hz, 8.8 Hz, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.16-7.11 (m, 1H), 7.01-6.94 (m,1H), 5.06 (d, J = 10.4 Hz, 1H), 4.34-4.29 (m, 1H), 3.99 (d, J = 2.4 Hz, 3H),2.80-2.76 (m, 1H), 1.67 (s, 3H), 0.83-0.80 (m, 3H).

[0156] Example 3 BX20-9-009

[0157] Step 1: Synthesis of intermediate S8

[0158] Reference Example 1 Step 1 to Step 9 Synthesis of Intermediate S8

[0159] Step 2: Synthesis of compound BX20-9-009

[0160]

[0161] Method: S8 (150 mg, 0.42 mmol) was dissolved in DCM:DMF (3 mL, 250:1) and (COCl) was added dropwise at 0 °C. 2 (160 mg, 1.26 mmol), the reaction solution was stirred at room temperature for 0.5 h. After the reaction, the reaction solution was concentrated under reduced pressure to obtain crude S9 (170 mg, crude), which was used directly in the next step. Int 9 (140 mg, crude, about 0.8 mmol) was dissolved in NMP (2 mL), TEA (91 mg, 0.9 mmol) was added, and then a DCM (2 mL) solution of S9 (180 mg, crude) was added dropwise, and stirred at room temperature for 1 hour. After the reaction, the solution was concentrated under reduced pressure and purified by preparative chromatography (neutral system), and freeze-dried to obtain a white solid BX20-9-009 (155 mg, 74.5%). MS (ESI, m / z) 498 [M+H] + .

[0162] 1 H NMR (400 MHz, CD 3 OD) δ = 8.57 (d, J = 2.0 Hz, 1H), 8.25 (s, 1H),8.15 (dd, J = 8.8 Hz, 2.4 Hz, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.15-7.11 (m,1H), 7.00-6.94 (m, 1H), 5.09 (d, J = 10.4 Hz, 1H), 4.35 (dd, J = 10.8 Hz, 8.0Hz, 1H), 3.99 (d, J = 2.4 Hz, 3H), 2.83-2.76 (m, 1H), 1.67 (s, 3H), 0.83-0.81(m, 3H).

[0163] Example 4 BX20-9-010

[0164] Step 1: Synthesis of intermediate S8

[0165] Reference Example 1 Step 1 to Step 9 Synthesis of Intermediate S8

[0166] Step 2: Synthesis of compound BX20-9-010

[0167] The synthetic route is shown below:

[0168]

[0169] Method: Compound S8 (0.15 g, 0.42 mmol) was dissolved in DMF (5 mL) and stirred until clear. HATU (0.24 g, 0.64 mmol) and TEA (0.17 g, 1.68 mmol) were added. After stirring for 5 min, Int 10 (0.10 g, 0.64 mmol) was added. The reaction mixture was stirred at 25 °C for 16 h. After the reaction, the reaction solution was added to saturated ammonium chloride to quench, extracted with EA, washed with concentrated brine, dried over anhydrous sodium sulfate, concentrated, and purified by preparative chromatography to obtain a white solid BX20-9-010 (42 mg, 20.1 %). MS (ESI, m / z) 498 [M+H] + .

[0170] 1 H NMR (400 MHz, CD 3 OD) δ 8.50 (d, J = 2.8 Hz, 1H), 8.05-8.04 (m, 1H), 8.02 (d, J = 2.4 Hz, 1H), 7.68 (d, J = 8.8 Hz, 1H), 7.19-7.14 (m, 1H), 7.03-6.96 (m, 1H), 5.10 (d, J = 10.4 Hz, 1H), 4.38-4.33 (m, 1H), 4.02 (d, J = 2.4Hz, 3H), 2.86-2.77 (m, 1H), 1.69 (s, 3H), 0.86-0.82 (m, 3H).

[0171] Example 5 BX20-9-025

[0172] Step 1: Synthesis of S8

[0173] Reference Example 1 Steps 1 to 9 to synthesize intermediate S8.

[0174] Step 2: Synthesis of intermediate 25-1

[0175] The synthetic route is shown below:

[0176]

[0177] Method: Compound S8 (150 mg, 0.423 mmol) was dissolved in DCM (3 mL), one drop of DMF was added, and 0.2 mL (COCl) was added dropwise under ice bath. 2 The reaction was stirred at room temperature for 0.5 hours. After the reaction was completed, the crude product S9 was obtained by concentrating under reduced pressure. Compound 7-aminoisoquinoline (60 mg, 0.42 mmol) was dissolved in NMP (3 mL), TEA (128 mg, 1.27 mmol) was added, and then the DCM solution of the crude product S9 was added dropwise, and the reaction was stirred at room temperature for 1 hour. After the reaction was completed, water was added to quench, DCM was extracted, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to obtain a brown solid 25-1 (200 mg, crude product).

[0178] Step 3: Synthesis of BX20-9-025

[0179] The synthetic route is shown below:

[0180]

[0181] Method: Compound 25-1 (200 mg, crude) was dissolved in DCM (10 mL), cooled to 0 °C, and m-CPBA (0.13 g, 0.63 mmol) was added. The reaction mixture was stirred at room temperature for 5 h. After the reaction, the reaction solution was added to 1N NaOH for quenching, extracted with DCM / MeOH, and the organic phases were combined, washed with concentrated brine, dried, concentrated under reduced pressure, and purified by preparative chromatography to obtain a white solid BX20-9-025 (83 mg, 39.8 %). MS (ESI, m / z) 497 [M+H] + .

[0182] 1 H NMR (400 MHz, DMSO-d6) δ 10.62 (s, 1H), 8.88 (d, J = 1.6 Hz, 1H), 8.29 (d, J = 2.0 Hz, 1H), 8.06-8.03 (m, 1H), 7.91–7.83 (m, 2H), 7.69-7.66 (m,1H), 7.22-7.13 (m, 2H), 5.14 (d, J = 10.4 Hz, 1H), 4.32-4.26 (m, 1H), 3.96(d, J = 2.0 Hz, 3H), 2.82-2.74 (m, 1H), 1.61 (s, 3H), 0.76-0.72 (m, 3H).

[0183] Example 6 BX20-9-026

[0184] Step 1: Synthesis of S8

[0185] Refer to Steps 1 to 9 of Example 1 to synthesize intermediate S8.

[0186] Step 2: Synthesis of intermediate S11

[0187] The synthesis route is shown as follows:

[0188]

[0189] Method: Dissolve S8 (150 mg, 0.424 mmol) in DCM (6 mL), add 1 drop of DMF, cool down to 0 °C, and slowly add dropwise (COCl) 2 (0.1 mL, 1.272 mmol), stir the reaction at room temperature for 0.5 hour. After the reaction is completed, concentrate under reduced pressure, then dissolve in DCM (5 mL), and slowly add dropwise to a DCM (6 mL) solution of A1 (74 mg, 0.509 mmol) and TEA (206 mg, 2.036 mmol), and stir at room temperature for 0.5 hour. After the reaction is completed, concentrate the reaction solution under reduced pressure, dissolve it with EA, add water (15 mL) dropwise to quench, extract with EA, combine the organic phases, wash with saturated NaCl solution, and dry with anhydrous Na 2 SO 4 Dry, concentrate under reduced pressure to obtain a yellow oily compound S11 (270 mg, crude product). MS (ESI, m / z) 481 [M+H] + .

[0190] Step 3: Synthesis of BX20-9-026

[0191] The synthesis route is shown as follows:

[0192]

[0193] Methods: Compound S11 (270 mg, crude, 0.424 mmol) was dissolved in DCM (6 mL), cooled to 0°C, and m-CPBA (128 mg, 0.636 mmol, 85% purity) was added. The reaction solution was stirred at room temperature for 4 hours. After the reaction, the reaction solution was added to 1 N NaOH (15 mL) for quenching, extracted with DCM / MeOH, the organic phases were combined, washed with saturated NaCl aqueous solution, dried, concentrated under reduced pressure, purified by preparative chromatography (formic acid system), and freeze-dried to obtain a white solid BX20-9-026 (95 mg, 45 %). MS (ESI, m / z) 497 [M+H] + .

[0194] 1 H NMR (400 MHz, DMSO-d6) δ 10.78 (s, 1H), 8.91 (d, J = 2.4 Hz, 1H), 8.29 (d, J = 2.0 Hz, 1H), 8.04 (d, J= 8.8 Hz, 1H), 7.91 (dd, J = 8.8 Hz, 2.0Hz, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.39-7.34 (m, 1H), 7.20-7.15 (m, 2H), 5.14(d, J = 10.4 Hz, 1H), 4.33-4.27 (m, 1H), 3.96 (d, J = 2.0 Hz, 3H), 2.83-2.75(m, 1H), 1.63 (s, 3H), 0.77-0.71 (m, 3H).

[0195] Example 7 BX20-9-028

[0196] Step 1: Synthesis of S8

[0197] Reference Example 1 Steps 1 to 9 to synthesize intermediate S8.

[0198] Step 2: Synthesis of intermediate 28-2

[0199] The synthetic route is shown below:

[0200]

[0201] Method: Compound 28-1 (4.6 g, 20 mmol) was dissolved in DMF (60 mL) and Cs 2 CO 3(13.0 g, 40 mmol) and acetone oxime (1.6 g, 22 mmol), the reaction solution was stirred at room temperature for 24 hours. After the reaction, EA extraction, brine washing, anhydrous sodium sulfate drying, and reduced pressure concentration to obtain a yellow oily liquid 28-2 (5.0 g, crude product). MS (ESI, m / z) 222 [M+H] + .

[0202] Step 3: Synthesis of intermediate 28-3

[0203]

[0204] Method: Add 28-2 (5.0, crude) to concentrated hydrochloric acid (15 mL), heat to 100 °C, and stir for 7 h. After the reaction, extract with EA, and concentrate the aqueous phase under reduced pressure to obtain a white solid powder 28-3 (1.0 g, crude). MS (ESI, m / z) 168[M+H] + .

[0205] Step 4: Synthesis of intermediate 28-4

[0206]

[0207] Method: 28-4 (1.0 g, crude product) was dissolved in acetic anhydride (8 mL) and stirred at 60 °C for 3 h. After the reaction, water was added to quench, EA was extracted, and NaHCO 3 The mixture was washed with water, washed with brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a white solid 28-4 (400 mg, 35.5%). MS (ESI, m / z) 214 [M+Na] + .

[0208] Step 5: Synthesis of intermediate 28-5

[0209]

[0210] Method: Cool fuming nitric acid (3 mL) to -30 °C, slowly add 28-4 (400 mg), stir the reaction solution at -30 °C for half an hour, then heat to 0 °C and stir for half an hour. After the reaction, add the reaction solution to crushed ice to quench, extract with EA, and then add NaHCO 3 The mixture was washed with water, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a light yellow solid 28-5 (400 mg, crude product). MS (ESI, m / z) 237 [M+H] + .

[0211] Step 6: Synthesis of intermediate 28-6

[0212]

[0213] Method: Dissolve 28-5 (400 mg, crude product) in EtOH (10 mL), add Pd / C (80 mg), replace hydrogen in the reaction system and stir at room temperature for 1 hour. After the reaction, filter through celite, and concentrate the filtrate under reduced pressure to obtain a yellow solid 28-6 (350 mg, crude product). MS (ESI, m / z) 207 [M+H] + .

[0214] Step 7: Synthesis of intermediate Int 28

[0215]

[0216] Method: Dissolve 28-6 (350 mg, crude) in dilute hydrochloric acid (5%, 4 mL) and stir at 50 °C for 30 minutes. After the reaction, concentrate under reduced pressure to obtain a yellow solid Int 28 (360 mg, crude). MS (ESI, m / z) 165 [M+H] + .

[0217] Step 8: Synthesis of BX20-9-028

[0218]

[0219] Method: S8 (150 mg, 0.42 mmol) was dissolved in DCM:DMF (3 mL, 250:1) and (COCl) was added dropwise at 0 °C. 2 (160 mg, 1.26 mmol), the reaction solution was stirred at room temperature for 0.5 h. After the reaction, the reaction solution was concentrated under reduced pressure to obtain crude S9 (180 mg, crude), which was used directly in the next step; Int 28 (168 mg, crude hydrochloride, about 0.8 mmol) was dissolved in NMP (2 mL), TEA (130 mg, 1.3 mmol) was added, and then a DCM (2 mL) solution of S9 (180 mg, crude) was added dropwise, and stirred at room temperature for 1 hour. After the reaction, the solution was concentrated under reduced pressure and purified by preparative chromatography, and freeze-dried to obtain a white solid BX20-9-028 (73 mg, 34.7%). MS (ESI, m / z) 501 [M+H] + .

[0220] 1 H NMR (400 MHz, DMSO-d 6) δ = 11.27 (s, 1H), 10.47 (s, 1H), 8.18 (d, J= 2.4 Hz, 1H), 7.80 (dd, J = 8.0 Hz, 2.4 Hz, 1H), 7.31 (d, J = 8.4 Hz, 1H),7.21-7.13 (m, 2H), 5.07 (d, J = 10.4 Hz, 1H), 5.01 (s, 2H), 4.25 (dd, J =10.4 Hz, 7.6 Hz, 1H), 3.95 (d, J = 2.0 Hz, 3H), 2.80-2.72 (m, 1H), 1.61 (s,3H), 0.72-0.74 (m, 3H).

[0221] Example 8 BX20-9-042

[0222] Step 1: Synthesis of S8

[0223] Reference Example 1 Steps 1 to 9 to synthesize intermediate S8.

[0224] Step 2: Synthesis of intermediate 42-2

[0225]

[0226] Method: Dissolve compound 42-1 (6.4 g, 32.8 mmol) in CCl 4 (60 mL), add BPO (75%, 800 mg, 2.5 mmol) and NBS (6.4 g, 35.9 mmol), and stir at 83 °C for 5 hours. After the reaction, filter, wash with sodium sulfite, wash with brine, dry and concentrate under reduced pressure to obtain yellow oily liquid 42-2 (8.6 g, crude product). MS (ESI, m / z) 274, 276 [M+H] + .

[0227] Step 3: Synthesis of intermediate 42-3

[0228]

[0229] Method: Compound 42-2 (4.3 g, crude product) was dissolved in 1,4-dioxane (50 mL), hydrazine hydrate (2.0 g, 40 mmol) was added, and the mixture was stirred at 100°C for 16 hours. After the reaction, the mixture was extracted with EA, washed with brine, dried, concentrated under reduced pressure, and purified by column chromatography to obtain yellow oily liquid 42-3 (500 mg, 16.5%). MS (ESI, m / z) 194[M+H] + .

[0230] Step 4: Synthesis of intermediate Int 42

[0231]

[0232] Method: Compound 42-3 (500 mg, 2.59 mmol) was dissolved in EtOH (30 mL), and Pd / C (10%, 100 mg) was added under nitrogen atmosphere. The reaction was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was filtered and concentrated under reduced pressure to obtain a yellow solid Int 42 (300 mg, 71.1%). MS (ESI, m / z) 164 [M+H] + .

[0233] Step 5: Synthesis of BX20-9-042

[0234]

[0235] Method: Compound S8 (150 mg, 0.42 mmol) was dissolved in DMF (5 mL), EDCI (161 mg, 0.84 mmol), HOBT (61 mg, 0.45 mmol) and DIEA (1.26 mmol, 162 mg) were added, and the mixture was stirred at room temperature for 5 minutes, and then Int 42 (65 mg, 0.4 mmol) was added. The mixture was stirred at room temperature for 3 hours. After the reaction, the mixture was extracted with EA, washed with brine, concentrated under reduced pressure, and purified by preparative chromatography to obtain a white solid BX20-9-042 (25 mg, 12.0%). MS (ESI, m / z) 500[M+H] + .

[0236] 1H NMR (400 MHz, DMSO-d6) δ 10.76-10.65 (m, 1H), 7.17-6.80 (m, 6H), 5.49 (d, J = 9.6 Hz, 1H), 4.95-4.66 (m, 2H), 4.30-4.05 (m, 1H), 3.88-3.79 (m,3H), 2.67-2.62 (m, 1H), 1.39-1.36 (m, 3H), 0.77-0.69 (m, 3H).

[0237] Example 9 BX20-9-045

[0238] Step 1: Synthesis of S8

[0239] Reference Example 1 Steps 1 to 9 to synthesize intermediate S8.

[0240] Step 2: Synthesis of intermediate 45-1

[0241]

[0242] Method: Compound 42-2 (4.3 g, crude product) was dissolved in 1,4-dioxane (50 mL), hydrazine hydrate (2.0 g, 40 mmol) was added, and the mixture was stirred at 100°C for 16 hours. After the reaction, the mixture was extracted with EA, washed with brine, dried, concentrated under reduced pressure, and purified by column chromatography to obtain yellow oily liquid 45-1 (400 mg, 13.2%). MS (ESI, m / z) 194[M+H] + .

[0243] Step 3: Synthesis of intermediate Int 45

[0244]

[0245] Method: Compound 45-1 (400 mg, 2.59 mmol) was dissolved in EtOH (30 mL), and Pd / C (10%, 100 mg) was added under nitrogen atmosphere. The reaction was stirred at room temperature for 1 hour. After the reaction was completed, the filtrate was filtered and concentrated under reduced pressure to obtain yellow solid Int 45 (238 mg, 70.6%). MS (ESI, m / z) 164 [M+H] + .

[0246] Step 4: Synthesis of BX20-9-045

[0247]

[0248] Method: Compound S8 (150 mg, 0.42 mmol) was dissolved in DMF (5 mL), EDCI (161 mg, 0.84 mmol), HOBT (61 mg, 0.45 mmol) and DIEA (1.26 mmol, 162 mg) were added, and the mixture was stirred at room temperature for 5 minutes, and then Int 45 (65 mg, 0.4 mmol) was added. The mixture was stirred at room temperature for 3 hours. After the reaction, the mixture was extracted with EA, washed with brine, concentrated under reduced pressure, and purified by preparative chromatography to obtain a white solid BX20-9-045 (16 mg, 7.7%). MS (ESI, m / z) 500[M+H] + .

[0249] 1 H NMR (400 MHz, DMSO-d6) δ 10.57 (s, 1H), 7.29-7.26 (m, 1H), 7.22-7.13 (m, 2H), 7.00-6.97 (m, 2H), 5.08 (d, J = 10.4 Hz, 1H), 4.34 (s, 2H), 4.15 (dd, J = 10.4 Hz, 7.6 Hz, 1H), 3.95 (d, J = 2.0 Hz, 3H), 2.76-2.67 (m,1H), 1.61 (s, 3H), 0.72-0.70 (m, 3H).

[0250] Example 10: Synthesis of compound BX20-9-050

[0251] Step 1: Synthesis of Intermediate 50-2

[0252]

[0253] Method: Compound 50-1 (2.1 g, 10 mmol) was dissolved in 1,4-dixoane (30 mL) and SeO 2 (1.33 g, 12 mmol), the reaction was stirred at 100 °C for 16 hours. After the reaction was completed, the filtrate was filtered, the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain a brown oily liquid 50-2 (2.0 g, 89.2%). MS (ESI, m / z) 225 [M+H] + .

[0254] Step 2: Synthesis of intermediate 50-3

[0255]

[0256] Method: Compound 50-2 (450 mg, 2 mmol) was dissolved in ethanol (5 mL), hydrazine hydrate (100 mg, 2 mmol) was added, and the reaction was stirred at 80 °C for 1 hour. After the reaction, water was added for extraction with EA, washed with brine, dried, concentrated under reduced pressure, and purified by column chromatography to obtain a yellow solid 50-3 (270 mg, 70.3%). MS (ESI, m / z) 193 [M+H] + .

[0257] Step 3: Synthesis of intermediate Int 50

[0258]

[0259] Method: Compound 50-3 (270 mg, 1.4 mmol) was dissolved in EtOH (10 mL) and THF (5 mL), and Pd / C (10%, 100 mg) was added under nitrogen atmosphere. The reaction system was replaced with hydrogen balloon and stirred at room temperature for 1 hour. After the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure to obtain yellow solid Int 50 (300 mg, 71.1%). MS (ESI, m / z) 163 [M+H] + .

[0260] Step 4: Synthesis of intermediate BX20-9-050

[0261]

[0262] Method: S8 (150 mg, 0.42 mmol) was dissolved in DCM (3 mL), one drop of DMF was added, and (COCl) was added dropwise at 0 °C. 2 (160 mg, 1.26 mmol), the reaction solution was stirred at room temperature for 0.5 h. After the reaction, the reaction solution was concentrated under reduced pressure to obtain crude S9 (180 mg, crude), which was used directly in the next step; Int 50 (65 mg, 0.4 mmol) was dissolved in NMP (2mL), TEA (130 mg, 1.3 mmol) was added, and then a DCM (2 mL) solution of S9 (180 mg, crude) was added dropwise, and stirred at room temperature for 0.5 hours. After the reaction, the solution was concentrated under reduced pressure and purified by preparative chromatography, and lyophilized to obtain a light yellow solid powder BX20-9-050 (36 mg, 17.2%). MS (ESI, m / z) 499 [M+H] + .

[0263] 11H NMR (400 MHz, DMSO-d6) δ = 12.87 (s, 1H), 10.90 (s, 1H), 9.24 (d, J = 2.4 Hz, 1H), 8.91 (d, J = 2.4 Hz, 1H), 8.35 (s, 1H), 7.22 - 7.14 (m, 1H), 5.18 (d, J = 10.4 Hz, 1H), 4.29 (dd, J 1 = 10.4 Hz, J 2 = 8.0 Hz, 1H), 3.95 (d, J = 2.4 Hz, 3H), 2.82 - 2.75 (m, 1H), 1.64 (s, 3H), 0.75 - 0.73 (m, 3H).

[0264] Biological Test Evaluation

[0265] The present invention will be further described and explained below in combination with test examples

[0266] Test Example 1 Blocking Activity of the Compounds of the Present Invention against Sodium Channel 1.8 (Nav1.8)

[0267] 1. Experimental Purpose: To detect the effects of compounds on the currents of voltage-gated sodium channel (NaV) subtypes 1.1 - 1.8 by using the patch clamp technique

[0268] 2. Experimental Materials and Instrumentation

[0269] 2.1. Cell Line: CHO cell line stably expressing Nav1.8 sodium channel. The Nav1.8 cells were self-constructed by the laboratory of Beijing Ace Medicine Co., Ltd. Gene Information: Sodium channel, voltage-gated, type 8, alpha (SCN10A), cDNA strictly similar to GenBank accession number: NM_006514

[0270] 2.2. Compounds: Dissolved in DMSO

[0271] 3. Experimental Method

[0272] 3.1. Cell Culture

[0273] (1) Maintenance medium: The cells were cultured in HAM'S / F-12 medium containing 10% fetal bovine serum, 10 μg / mL Blasticidin, 200 μg / mL Hygromycin B, and 100 μg / mL Zeocin at 37°C and a carbon dioxide concentration of 5%.

[0274] (2) Cell passaging: Remove the old culture medium and wash once with PBS, then add 1 mL of 0.25%-Trypsin-EDTA solution and incubate at 37°C for about 1.5 min. When the cells detach from the bottom of the dish, add about 5 mL of complete culture medium preheated at 37°C. Gently blow the cell suspension with a pipette to separate the aggregated cells. Transfer the cell suspension to a sterile centrifuge tube and collect the cells by centrifugation at 1000 rpm for 5 min. For expansion or maintenance culture, inoculate the cells in a 6 cm cell culture dish with 2.5×105 cells in each cell culture dish (final volume: 5 mL).

[0275] (3) To maintain the electrophysiological activity of cells, the cell density must not exceed 80%.

[0276] (4) Patch clamp assay: Before the test, cells were detached with 0.25%-Trypsin-EDTA, 6.5×103 cells were plated on coverslips and cultured in 24-well plates (final volume: 500 µL). After 18 hours, the test was performed.

[0277] 3.2. Patch clamp assay

[0278] (1) After the whole-cell seal is formed, the cell voltage is clamped at -120 mV. First, the voltage is stepped from -130 mV to -10 mV in 10 mV steps and maintained for 5 s, and then a 0 mV depolarization pulse is given to obtain the half-inactivated voltage (Vhalf). The resting state and half-inactivated state of the sodium current are detected using a double pulse mode. First, the first depolarization pulse (TP1) is given to 0 mV for 50 ms to detect the sodium current in the resting state. Then the voltage is adjusted to Vhalf and maintained for 5 s, and then the voltage is restored to -120 mV and maintained for 20 ms. Then a second depolarization pulse (TP2) is given to 0 mV for 50 ms to detect the sodium current in the half-inactivated state. Finally, it is restored to the clamping voltage of -120 mV. Data is collected repeatedly every 20 ms to observe the effect of the drug on the peak sodium current in the two different states. The test data were collected by EPC10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0279] (2) When performing the patch clamp operation, first use a microelectrode puller to pull the capillary glass tube into a recording electrode, then place the electrode filled with intracellular fluid into the microelectrode holder, and manipulate the microelectrode manipulator under an inverted microscope to bring the recording electrode into contact with the cell, applying negative pressure to form a GΩ seal. At this time, perform fast capacitance compensation, and then continue to apply negative pressure to break the cell membrane and form a whole-cell recording mode. Finally, perform slow capacitance compensation and record related parameters. No leakage compensation is given.

[0280] (3) When the sodium current recorded by the whole cell is stable, the drug is administered. Each drug concentration is applied for 5 min (or the current is stable) before the next concentration is detected. The coverslip with cells is placed in the recording bath under an inverted microscope. The blank control external solution and the working solution of the compound to be tested flow through the recording bath from low concentration to high concentration by gravity perfusion to act on the cells. A peristaltic pump is used for liquid exchange during recording. The current detected by each cell in the external solution without the compound serves as its own control group. Each concentration is tested twice independently. All electrophysiological experiments are performed at room temperature.

[0281] Data analysis

[0282] First, the current after each drug concentration and the blank control current were normalized, and then the inhibition rate corresponding to each drug concentration was calculated, that is, (1-compound action current / blank control current), and the mean (Mean), standard deviation (SD) and standard error (SE) of the inhibition rate of each concentration were calculated, and the data were expressed as mean ± SE.

[0283] 4. Experimental Results

[0284] Table: Blocking rate of the compounds of the present invention on NaV1.8 at 100 nM

[0285]

[0286] It can be seen that the compounds of the present invention have a significant blocking effect on the activity of NaV1.8 channels.

Claims

1. A compound as shown in formula I or a pharmaceutically acceptable salt thereof, ； in, R 1 , R 2 and R 3 Each is independently halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, -OR 1-1 , by one or more R 1-2 Substituted C1-C6 alkyl or one or more R 1-3 Substituted C1-C6 alkoxy; R 1-1 is a C3-C6 cycloalkyl group or a 3-8 membered heterocycloalkyl group; in the 3-8 membered heterocycloalkyl group, the heteroatoms are selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3; Each R 1-2 are each independently a halogen; Each R 1-3 Each is independently a C1-C6 alkoxy group; Ring A is C6-C 10 Aryl or 5-10 membered heteroaryl; in the 5-10 membered heteroaryl, the heteroatoms are selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3; Ring B is a 5-6 membered heterocycloalkyl, a 5-6 membered heterocycloalkenyl or a 5-6 membered heteroaryl; in the 5-6 membered heterocycloalkyl, the 5-6 membered heterocycloalkenyl and the 5-6 membered heteroaryl, the heteroatoms are selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3; n is 0, 1, 2, 3, 4 or 5; Each R 4 Each is independently oxo, -NH2, halogen or -OH; R 7 is hydrogen or C1-C6 alkyl; R 8 It is hydrogen, C1-C6 alkyl, C1-C6 alkyl-C1-C6 alkoxy, C1-C6 alkoxy or -OH.

2. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: It meets one or more of the following conditions: (1) R 1 , R 2 and R 3 wherein the halogen is independently fluorine, chlorine, bromine or iodine, preferably fluorine; (2) R 1 , R 2 and R 3 wherein the C1-C6 alkyl group and the 1-2 The C1-C6 alkyl groups in the substituted C1-C6 alkyl groups are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, preferably methyl; (3) R 1 , R 2 and R 3 wherein the C3-C6 cycloalkyl groups are each independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, preferably cyclopropyl; (4) R 1 , R 2 and R 3 wherein the C1-C6 alkoxy group and the 1-3 The C1-C6 alkoxy groups in the substituted C1-C6 alkoxy groups are each independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy, preferably methoxy or ethoxy; (5) R 1-1 In the above, the C3-C6 cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, preferably cyclopropyl; (6) R 1-1 wherein the heteroatom in the 3-8 membered heterocycloalkyl is N and / or O; the number of heteroatoms is preferably 1 or 2; the 3-8 membered heterocycloalkyl is preferably a 4-6 membered heterocycloalkyl; more preferably ;For example ; (7) R 1-2 wherein the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine; (8) R 1-3 wherein the C1-C6 alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy, preferably methoxy; (9) In ring A, the C6-C 10 Aryl is phenyl or naphthyl; preferably phenyl; (10) In ring A, the heteroatom of the 5-10 membered heteroaryl group is N and / or O; the number of heteroatoms is preferably 1 or 2; the 5-10 membered heteroaryl group is preferably a 5-6 membered heteroaryl group; more preferably ; (11) In ring B, the heteroatom in the 5-6 membered heterocycloalkyl is N and / or O; the number of heteroatoms is preferably 1 or 2; the 5-6 membered heterocycloalkyl may be a 5 membered heterocycloalkyl or a 6 membered heterocycloalkyl; preferably , , or ; (12) In ring B, the heteroatom in the 5-6-membered heterocycloalkenyl group is N and / or O; the number of heteroatoms is preferably 1 or 2; the 5-6-membered heterocycloalkenyl group may be a 5-membered heterocycloalkenyl group or a 6-membered heterocycloalkenyl group; preferably , or ; (13) In ring B, the heteroatom in the 5-6 membered heteroaryl group is N and / or O; the number of heteroatoms is preferably 1 or 2; the 5-6 membered heteroaryl group is a 5-membered heteroaryl group or a 6-membered heteroaryl group; preferably or ; (14) R 4 wherein the halogen is fluorine, chlorine, bromine or iodine, preferably fluorine; (15) R 4 When the same carbon atom is occupied by two R 4 When substituted, the oxo group is =O; that is, together with the attached C, it forms C=O; and (16) R 4 When the N atom is replaced by an R 4 When substituted, the oxo is -O - ; That is, together with the connected N, it forms N + -O - .

3. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: It meets one or more of the following conditions: (1) R 1 is a C1-C6 alkoxy group; (2) R 2 is a halogen; (3) R 3 is a halogen; (4) n is 0, 1, 2 or 3; (5) When R 4 When located on ring A, each R 4 are each independently halogen; and (6) When R 4 When located on ring B, each R 4 Each is independently oxo, -NH2 or -OH.

4. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: It is the following scheme 1 or scheme 2: Scheme 1: The compound represented by formula I is a compound represented by the following formula I-1: ； Among them, R 1 , R 2 , R 3 , n and ring A are defined as any one of claims 1 to 3; X 1 and X 2 are each independently selected from C or N; X 3 , X 4 , X 5 and X 6 Each independently selected from: chemical bond, O, S, S(O), S(O2), N, NH, CH, NR 4 , CR 4 or C(R 4 )2; and X 3 , X 4 , X 5 and X 6 At most one of them is a chemical bond at a time; Dashed lines indicate chemical bonds or none; R 4 is oxo, -NH2, halogen or -OH; Scheme 2: The compound shown in Formula I is a compound shown in Formula I-3 below: Among them, R 1 , R 2 , R 3 , R 4 , Ring A, Ring B and n are as defined in any one of claims 1 to 3.

5. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 4, characterized in that: The compound represented by formula I-1 is a compound represented by formula I-2 below, or the compound represented by formula I-3 is a compound represented by formula I-4 below: ； ； Among them, ring A, ring B, R 4 , n, X 1 , X 2 , X 3 , X 4 , X 5 and X 6 The definition as set forth in claim 4.

6. The compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: It meets one or more of the following conditions: (1) R 1 is methoxy; (2) R 2 For fluorine; (3) R 3 For fluorine; (4) R 7 is methyl; (5) R 8 is methyl; (6) Ring A is or ; (7) Ring B is , , , , , , or ; (8) n is 1, 2 or 3; and (9) R 4 is oxo, -OH, -NH2 or F; Preferably, the compound of formula I or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 3 satisfies the following conditions: for , , , , , , , , , or .

7. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 6, characterized in that: The compound as shown in formula I is any of the following compounds: 。 8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (1) a compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, and (2) Pharmaceutically acceptable excipients.

9. Use of a substance A in the preparation of a medicament for treating a disease; the disease may be pain, a pain-related disease, multiple sclerosis, incontinence or arrhythmia; the pain is preferably one or more of acute pain, chronic pain, inflammatory pain, cancer pain, neuropathic pain, musculoskeletal pain, primary pain, intestinal pain and idiopathic pain; Alternatively, the disease may be a disease for treatment by inhibiting voltage-gated sodium channels; the voltage-gated sodium channels are preferably Na V 1.8; The substance A is a compound as shown in formula I or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 7, or a pharmaceutical composition as described in claim 8.

10. A use of the substance A in the preparation of a voltage-gated sodium channel inhibitor; the voltage-gated sodium channel is preferably Na V 1.8; The substance A is as described in claim 9.

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