Pharmaceutical compounds targeting stings and methods of making and using the same

Abstract

The application discloses a kind of medicinal compounds with STING as target point and preparation method and purposes thereof, structure is as shown in general formula I, it also includes the enantiomeric isomer, diastereoisomer, racemate and pharmaceutically acceptable salt of compound in formula I, metabolite, prodrug, solvate or hydrate.This application overcomes the defects of the lack of existing STING small molecule inhibitor species, the benzoheterocyclic compound in the application can be used as STING regulator, and can also be used for preparing the drug for treating autoimmune diseases.

Description

[0001] The present application is a divisional application, the original case of which has a patent number of 2023104189666, an application date of April 19, 2023, and an invention title of Medicinal compound targeting STING and preparation method and use thereof. TECHNICAL FIELD

[0002] The present application relates to a class of medicinal compounds, in particular to a class of medicinal compounds targeting STING, and relates to their preparation method and application. The present application also relates to the use of the compounds. BACKGROUND

[0003] The innate immunity and adaptive immunity together constitute the human immune system. The innate immunity is the immunity with which the body is born and is the first line of defense of the body's immunity. The pattern recognition receptors (PRRs) expressed on the cell membrane and in the cytoplasm activate the innate immunity by recognizing the pathogen-associated molecular patterns (PAMPs) of non-self pathogens, and then kill and remove the invading pathogenic microorganisms such as viruses, bacteria and parasites.

[0004] Among them, the stimulator of interferon gene (STING) is a PRR of cyclic dinucleotides (CDNs). The invasion of pathogenic microorganisms and cell damage cause abnormal aggregation of DNA in the cytoplasm, and the cyclic GMP-AMP synthase (cGAS) catalyzes the generation of cyclic dinucleotides by recognizing double-stranded DNA in the cytoplasm of exogenous substances or self, activates STING and makes STING transfer from the endoplasmic reticulum to the Golgi body, induces the secretion of type I interferon (IFN), expresses and secretes inflammatory factors to regulate the immune response of the body, activates T cells and thus plays an anti-tumor immune role.

[0005] The development of STING modulators has received extensive attention from major pharmaceutical companies and research institutions at home and abroad in recent years. The currently reported STING agonists can be divided into cyclic dinucleotides and their analogs, xanthone and acridone, benzimidazole, and benzothiophene. They play an important role in tumor immunotherapy. Among them, the development of cyclic dinucleotide agonists is relatively early, but such compounds have defects such as large relative molecular mass, high molecular polarity, and poor cell membrane permeability, and the prospects for drug development are not ideal. In addition, inhibitors targeting STING can bring new hope for the treatment of autoimmune diseases, but there is no small molecule new drug targeting STING on the market.

[0006] Chinese invention patent application with publication number CN114805309A discloses a "benzoheterocyclic compound, its preparation method, pharmaceutical composition and application", which claims to effectively increase the release amount of interferon beta (IFN-β) at the cellular level, with the optimal release level reaching the level of hundreds of picomolar concentration, and can be used for preparing drugs for treating diseases such as tumors, especially drugs for tumor immunotherapy. Such compounds have poor drug properties due to large lipophilicity (LogP greater than 4). SUMMARY

[0007] The technical problem to be solved by the present application is to provide a class of benzoheterocyclic compounds or their pharmaceutically acceptable salts, isomers, preparation methods, pharmaceutical compositions, and their applications in anti-tumor and infectious diseases, which have the function of regulating the interferon gene stimulator protein, in view of the defects of single structure and limited efficacy of existing interferon gene stimulator protein modulators.

[0008] The technical solution of the present application is as follows:

[0009] The pharmaceutical compound targeting STING is characterized in that the compound is a benzoheterocyclic compound or any one of the pharmaceutically acceptable salts of the following structural formula:

[0010] A-1:

[0011] B-3:

[0012] B-4:

[0013] B-7:

[0014] B-8:

[0015] B-9:

[0016] B-10:

[0017] B-11:

[0018] B-12:

[0019] C-4:

[0020] D-1:

[0021] D-3:

[0022] D-4:

[0023] F-2:

[0024] F-3:

[0025] F-4:

[0026] F-7:

[0027] F-8:

[0028] F-11:

[0029] F-14:

[0030] Preferably, when the benzoheterocycle compound is used as a pharmaceutical salt, the salt is an acid salt of at least one of the following acids: alkyl sulfate, benzenesulfonic acid, citric acid, glycolic acid, lactic acid, malic acid, malonic acid, mandelic acid, phosphoric acid, hydrochloric acid, sulfuric acid, tartaric acid, succinic acid, formic acid, hydroiodic acid, hydrobromic acid, methanesulfonic acid, nicotinic acid, nitric acid, orotic acid, oxalic acid, pyroglutamic acid, saccharin acid, salicylic acid, gentisic acid, p-toluenesulfonic acid, valeric acid, palmitic acid, stearic acid, cinnamic acid, carbonic acid, ethanedisulfonic acid, ethylsuccinic acid, and fumaric acid.

[0031] The preparation method of the pharmaceutical compound targeting STING, the preparation reaction formula of the B-1 compound is as follows:

[0032]

[0033] The X substituent group corresponds to the substituent group on the B-1 compound.

[0034] The preparation method of the pharmaceutical compound targeting STING, the preparation reaction formula of the B-3 compound is as follows:

[0035]

[0036] wherein the R1 substituent corresponds to the substituent on the B-3 compound.

[0037] The preparation method of the medicinal compound targeting STING is as follows:

[0038]

[0039] wherein the R1, R2 substituent corresponds to the substituent on the D-3 or F-4 or F-7 or F-8 compound.

[0040] The benzo-heterocyclic compound of the present application targets STING, and has the pharmaceutical prospect of treating and / or preventing inflammatory diseases and autoimmune diseases. As a novel small molecule compound, the benzo-heterocyclic compound of the present application has higher inhibitory activity on interferon gene stimulatory protein, simple structure, convenient synthesis, and high safety. DETAILED DESCRIPTION

[0041] The technical solutions of the present application are further described in detail below in combination with examples. In the present application, the examples described below are to better illustrate the present application, and are not to limit the scope of the present application. Various modifications and changes can be made to the present application without departing from the present application.

[0042] The raw materials and equipment used in the specific embodiments of the present application are known products. The starting materials in the present application can be synthesized by known synthesis methods in the art, or obtained by purchasing commercially available conventional reagents, mainly purchased from Aladdin, Bide Pharmaceutical, and Leyen companies. The specific contents are shown in the following table.

[0043]

[0044]

[0045] The structure of the compound in the present application is determined by nuclear magnetic resonance (NMR). The determination solvent is deuterated chloroform (Chloroform-d) or deuterated dimethyl sulfoxide (DMSO-d6), and the internal standard is tetramethylsilane (TMS).

[0046] The reaction progress detection in the examples uses thin layer chromatography (TLC), and the developing agents used include petroleum ether / ethyl acetate system and dichloromethane / methanol system. The volume ratio of each solvent in the developing agent is adjusted according to the polarity of the compound.

[0047] The column chromatography was used to purify the compounds in the examples, and the eluents used included petroleum ether / ethyl acetate system; dichloromethane / methanol system. The volume ratio of each solvent in the eluent was adjusted according to the polarity of the compound.

[0048] The thin layer chromatography silica gel plate used was Yantai Huanghai HSGF254 silica gel plate, with a size of 75*25mm.

[0049] The column chromatography generally used silica gel 200-300 mesh silica gel from Qingdao Haoyang Chemical Factory Branch as the carrier.

[0050] The present specification uses the abbreviations in the following table:

[0051]

[0052]

[0053] Example 1: Synthesis of compound A-1

[0054]

[0055] Step 1: Under nitrogen protection, monomethyl malonate (0.33ml, 3.15mmol, 1.1eq) and EDCI (906mg, 4.72mmol, 1.5eq), HOBT (639mg, 4.72mmol, 1.5eq), Et3N (0.87ml, 6.30mmol, 2eq) were added to 10ml DCM, and the reaction mixture was stirred at room temperature under nitrogen atmosphere for 30min; then 2-aminoquinoline (500mg, 3.46mmol, 1.1eq) was added, and the reaction mixture was continuously stirred at room temperature under nitrogen atmosphere for 8-10 hours. The reaction process was monitored by TLC (V petroleum ether:V ethyl acetate=4:1), and when the starting material no longer decreased, the reaction was stopped. After the reaction was completed, the DCM was concentrated, the reaction system was extracted with EA (3*30ml), and the organic phase was washed with H2O (2*30ml), saturated brine (1*30ml) in sequence, and the combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product after concentration was purified by silica gel column chromatography (PE:EA=10:1) to obtain yellow solid, which was compound 1b (631mg, 81%). 1H NMR (400 MHz, Chloroform-d) δ 9.63 (s, 1H), 8.35 (s, 1H), 8.17 (d, J = 8.9 Hz, 1H), 7.87 (d, J = 8.5 Hz, 1H), 7.78 (d, J = 8.1 Hz, 1H), 7.66 (ddd, J = 8.4, 6.9, 1.4 Hz, 1H), 7.46 (ddd, J = 8.1, 7.0, 1.1 Hz, 1H), 3.82 (d, J = 1.8 Hz, 3H), 3.56 (s, 2H).

[0056]

[0057] Step 2: Compound 1b (300 mg, 1.22 mmol, 1 eq) was added to 5 ml MeOH and 5 ml H2O system stirring dissolution, then added NaOH (245 mg, 6.14 mmol, 5 eq); the reaction mixture was heated at 60°C for 1-1.5 hours. TLC (V petroleum ether: V ethyl acetate = 3:1) monitoring reaction process, when the raw material no longer reduced, the reaction was stopped. After the reaction was completed, the reaction solution was concentrated, the reaction mixture was extracted with EA (1x30 ml), washed with saturated NaHC03 aqueous solution (3x30 ml), the combined aqueous phase was adjusted to pH 2-3 with 10% HC1, and the yellow solid was precipitated overnight. The solid was filtered and dried to obtain compound A-1 (230 mg, 81%). 1 H NMR (400 MHz, DMSO-d6) δ 7.89 (d, J = 8.8 Hz, 1H), 7.63 (d, J = 7.9 Hz, 1H), 7.51 - 7.43 (m, 2H), 7.15 (ddd, J = 8.0, 5.2, 2.9 Hz, 1H), 6.77 (d, J = 8.8 Hz, 1H), 6.44 (s, 2H).

[0058] Example 2: Synthesis of compound B-1

[0059]

[0060] Step 1: 6-hydroxy-1-naphthoic acid (300 mg, 1.22 mmol, 1 eq) was dissolved in 10 ml DCM, and then imidazole (361 mg, 5.31 mmol, 2 eq), TBSCL (569 mg, 3.95 mmol, 1.49 eq) were added successively under stirring; the reaction mixture was stirred at room temperature for 3-4.5 hours. The reaction process was monitored by TLC (V petroleum ether:V ethyl acetate = 4:1), and the reaction was stopped when the starting material was no longer reduced. After the reaction was completed, the reaction solution was concentrated, and the reaction mixture was extracted with DCM (1 x 30 ml), washed with saturated NaHC03 aqueous solution (1 x 30 ml), and dried over anhydrous Na2S04. After filtration, the crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (PE:EA = 8:1) to obtain white solid 3b (580 mg, 76%). 1 H NMR (400 MHz, Chloroform-d) δ 8.65 (s, 1H), 8.07 (dd, J = 8.6, 1.4 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.75 (d, J = 8.8 Hz, 1H), 7.23 (d, J = 2.2 Hz, 1H), 7.16 (dd, J = 8.8, 2.4 Hz, 1H), 1.04 (s, 9H), 0.29 (s, 6H).

[0061]

[0062] Step 2: Under nitrogen protection, 3b (322 mg, 1.12 mmol, 1 eq) and EDCI (323 mg, 1.68 mmol, 1.5 eq), HOBT (227 mg, 1.68 mmol, 1.5 eq), DIPEA (0.37 ml, 2.24 mmol, 2 eq) were added to 10 ml DCM, and the reaction mixture was stirred at room temperature for 30 min under nitrogen atmosphere; then glycine methyl ester hydrochloride (155.24 ml, 1.23 mmol, 1.1 eq) was added, and the reaction mixture was continuously stirred at room temperature for 8-10 hours under nitrogen atmosphere. The reaction process was monitored by TLC (V petroleum ether:V ethyl acetate = 2:1), and the reaction was stopped when the starting material was no longer reduced. After the reaction was completed, the DCM was concentrated, the reaction system was extracted with EA (3 x 30 ml), and the organic phase was washed successively with H20 (2 x 30 ml) and saturated brine (1 x 30 ml). The combined organic phase was dried over anhydrous Na2S04, filtered, and the crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (PE:EA = 7:1) to obtain colorless oil, i.e. compound 3c (318 mg, 79%). 1H NMR (400 MHz, DMSO-d6) δ 8.11 - 8.03 (m, 2H), 7.73 (dd, J = 9.0, 2.5 Hz, 1H), 7.39 - 7.32 (m, 2H), 6.49 (s, 1H), 2.89 (s, 1H), 2.72 (d, J = 0.6 Hz, 1H).

[0063]

[0064] Step 3: Compound 3c (300 mg, 0.83 mmol, 1 eq) was dissolved in THF, then TBAF (0.345 ml, 1.25 mmol, 1.5 eq) was added, the reaction mixture was stirred at room temperature for 2-2.5 hours. TLC (V petroleum ether: V ethyl acetate = 1:1) was used to monitor the reaction process, and the reaction was stopped when the raw material was reacted completely. After the reaction was completed, the THF was concentrated, the reaction system was extracted with EA (3 x 30 ml), the organic phase was washed with H2O (2 x 30 ml) and saturated brine (1 x 30 ml) in turn, the combined organic phase was dried over anhydrous Na2SO4, filtered, and the crude product after concentration under reduced pressure was purified by silica gel column chromatography (PE:EA = 5:1) to obtain white solid, which was compound 3d (200 mg, 92%). 1 H NMR (400 MHz, DMSO-d6) δ 10.05 (s, 1H), 8.94 (t, J = 5.8 Hz, 1H), 8.37 - 8.28 (m, 1H), 7.90 - 7.65 (m, 3H), 7.19 - 7.06 (m, 2H), 4.01 (d, J = 5.8 Hz, 2H), 3.63 (s, 3H).

[0065] Step 4: According to the method of Example 1, replace 1b with 3d to prepare compound B-1 (190 mg, 77%). 1 H NMR (400 MHz, DMSO-d6) δ 10.05 (s, 1H), 8.94 (t, J = 5.8 Hz, 1H), 8.37 - 8.28 (m, 1H), 7.90 - 7.65 (m, 3H), 7.19 - 7.06 (m, 2H), 4.01 (d, J = 5.8 Hz, 2H), 3.63 (s, 3H).

[0066] Example 3: Synthesis of compound B-3

[0067]

[0068]

[0069] Step 1: 2-Fluoro-4-methoxybenzaldehyde (1.0 g, 6.49 mmol, 1.0 eq) was dissolved in 10 ml DMF, ethyl mercaptoacetate (0.920 ml, 8.43 mmol, 1.2 eq), potassium carbonate (2.69 g, 19.5 mmol, 3 eq) were added, and the temperature was raised to 70 °C. The reaction mixture was stirred at 70 °C for 1.5-3 h. The reaction process was monitored by TLC (V petroleum ether: V ethyl acetate = 30:1), and the reaction was stopped when the starting material was no longer reduced. After the reaction was completed, the DMF was concentrated, and the reaction system was extracted with EA (3 x 30 ml). The organic phase was washed successively with H2O (2 x 30 ml), saturated brine (1 x 30 ml), and then dried over anhydrous Na2SO4. After filtration, the organic phase was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA = 30:1) to obtain white solid, which was compound 5b (1.4 g, 91%). 1 H NMR (400 MHz, Chloroform-d) δ 7.95 (s, 1H), 7.72 (d, J = 8.81 Hz, 1H), 7.27 (d, J = 2.34 Hz, 1H), 7.01 (dd, J = 8.84, 2.35 Hz, 1H), 4.38 (q, J = 7.13 Hz, 3H), 3.88 (s, 3H), 1.39 (t, J = 7.13 Hz, 4H).

[0070]

[0071] Step 2: Compound 5b (1.4 g, 5.93 mmol, 1 eq) was added to a system of 5 ml MeOH and 5 ml H2O, stirred to dissolve, and then NaOH (593 mg, 14.8 mmol, 2.5 eq) was added. The reaction mixture was heated at 60 °C for 1-1.5 h. The reaction process was monitored by TLC (V petroleum ether: V ethyl acetate = 3:1), and the reaction was stopped when the starting material was no longer reduced. After the reaction was completed, the reaction liquid was concentrated, and the reaction mixture was extracted with EA (1 x 30 ml) and saturated NaHCO3 aqueous solution (3 x 30 ml). The combined aqueous phase was washed with 10% HCl to adjust the pH to 2-3, and then left to stand overnight. White solids were precipitated, which were filtered and dried to obtain compound 5c (1.1 g, 89%). 1 H NMR (400 MHz, DMSO-d6) δ 13.20 (s, 1H), 7.97 (s, 1H), 7.84 (d, J = 8.80 Hz, 2H), 7.56 (d, J = 2.38 Hz, 2H), 7.03 (dd, J = 8.82, 2.38 Hz, 1H), 3.81 (s, 3H).

[0072]

[0073] Step 3: Compound 5c (300 mg, 1.44 mmol, 1 eq) and EDCI (414 mg, 2.16 mmol, 1.5 eq), HOBT (292 mg, 2.16 mmol, 1.5 eq), DIPEA (0.480 ml, 2.88 mmol, 2 eq) were added to 10 ml DCM, the reaction mixture was stirred at room temperature for 30 min under nitrogen atmosphere; then glycine methyl ester hydrochloride (200 mg, 1.58 mmol, 1.1 eq) was added, the reaction mixture was continuously stirred at room temperature for 8-10 hours under nitrogen atmosphere. The reaction process was monitored by TLC (V dichloromethane: V methanol = 20: 1), when the raw material was no longer reduced, the reaction was stopped. After the reaction was completed, the DCM was concentrated, the reaction system was extracted with EA (3 x 30 ml), the organic phase was washed with H2O (2 x 30 ml), saturated brine (1 x 30 ml) in turn, the combined organic phase was dried over anhydrous Na2SO4, filtered, and the crude product after concentration under reduced pressure was purified by silica gel column chromatography (PE: EA = 2: 1) to obtain white solid, i.e. compound 5d (391 mg, 92%). 1 H NMR (400 MHz, Chloroform-d) δ 7.71 (s, 1H), 7.68 (d, J = 8.81 Hz, 1H), 7.26 (d, J = 2.34 Hz, 1H), 7.00 (dd, J = 8.81, 2.34 Hz, 1H), 6.65 (t, J = 4.81 Hz, 1H), 4.24 (d, J = 5.12 Hz, 2H), 3.87 (s, 3H), 3.79 (s, 3H).

[0074]

[0075] Step 4: Compound 5d (328 mg, 1.17 mmol, 1 eq) was added to 3 ml MeOH and 3 ml H2O system, stirred to dissolve, then NaOH (117 mg, 2.94 mmol, 2.5 eq) was added; the reaction mixture was heated at 60°C for 1-1.5 hours. The reaction process was monitored by TLC (V dichloromethane: V methanol = 20: 1), when the raw material was no longer reduced, the reaction was stopped. After the reaction was completed, the reaction liquid was concentrated, the reaction mixture was extracted with EA (1 x 30 ml), and the saturated NaHCO3 aqueous solution (3 x 30 ml) was washed, the combined aqueous phase was adjusted to PH 2-3 with 10% HCl, and the white solid was precipitated after standing overnight, and the solid was filtered and dried to obtain compound B-3 (246 mg, 79%). 1H NMR (400 MHz, DMSO-d6) δ 12.68 (s, 1H), 8.95 (t, J = 5.93 Hz, 1H), 7.98 (s, 1H), 7.80 (d, J = 8.81 Hz, 1H), 7.55 (d, J = 2.36 Hz, 2H), 7.01 (dd, J = 8.80, 2.39 Hz, 1H), 3.89 (d, J = 5.94 Hz, 2H), 3.80 (s, 3H).

[0076] The synthesis of compounds B-4 to B-16 in Examples 6 to 18 below can be carried out according to the synthetic method of Example 5, only by changing the corresponding starting materials.

[0077] Example 4: Synthesis of compound B-4

[0078]

[0079] Compound B-4 was characterized by structural parameters as follows: 1 H NMR (400 MHz, DMSO-d6) δ 12.56 (s, 1H), 8.80 (d, J = 7.28 Hz, 1H), 8.06 (s, 1H), 7.79 (d, J = 8.77 Hz, 1H), 7.55 (d, J = 2.37 Hz, 1H), 7.01 (dd, J = 8.79, 2.38 Hz, 1H), 4.40-4.32 (m, 1H), 3.80 (s, 3H), 1.37 (d, J = 7.35 Hz, 3H).

[0080] Example 5: Synthesis of compound B-7

[0081]

[0082] Compound B-7 was characterized by structural parameters as follows: 1 H NMR (400 MHz, DMSO-d6) δ 12.62 (s, 1H), 8.87 (d, J = 7.27 Hz, 1H), 8.10 (s, 1H), 7.80 (d, J = 8.21 Hz, 1H), 7.76 (s, 1H), 7.23 (d, J = 9.17 Hz, 1H), 4.50-4.27 (m, 1H), 2.40 (s, 3H), 1.37 (d, J = 7.35 Hz, 3H).

[0083] Example 6: Synthesis of compound B-8

[0084]

[0085] Compound B-8 was characterized by structural parameters as follows: 1H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.87 (d, J = 7.26 Hz, 1H), 8.10 (s, 1H), 7.80 (d, J = 8.22 Hz, 1H), 7.76 (s, 1H), 7.23 (dd, J = 1.05, 8.18 Hz, 1H), 4.41 ~ 4.33 (m, 1H), 2.40 (s, 3H), 1.37 (d, J = 7.35 Hz, 3H).

[0086] Example 7: Synthesis of compound B-9

[0087]

[0088] Compound B-9 was characterized by structural parameters: 1 H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.87 (d, J = 7.26 Hz, 1H), 8.10 (s, 1H), 7.80 (d, J = 8.22 Hz, 1H), 7.76 (s, 1H), 7.23 (dd, J = 1.05, 8.18 Hz, 1H), 4.41 ~ 4.33 (m, 1H), 2.40 (s, 3H), 1.37 (d, J = 7.35 Hz, 3H).

[0089] Example 8: Synthesis of compound B-10

[0090]

[0091] Compound B-10 was characterized by structural parameters: 1 H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.87 (d, J = 7.26 Hz, 1H), 8.10 (s, 1H), 7.80 (d, J = 8.22 Hz, 1H), 7.76 (s, 1H), 7.23 (dd, J = 1.05, 8.18 Hz, 1H), 4.41 ~ 4.33 (m, 1H), 2.40 (s, 3H), 1.37 (d, J = 7.35 Hz, 3H).

[0092] Example 9: Synthesis of compound B-11

[0093]

[0094] Compound B-11 was characterized by structural parameters: 1 H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.87 (d, J = 7.26 Hz, 1H), 8.10 (s, 1H), 7.80 (d, J = 8.22 Hz, 1H), 7.76 (s, 1H), 7.23 (dd, J = 1.05, 8.18 Hz, 1H), 4.41 ~ 4.33 (m, 1H), 2.40 (s, 3H), 1.37 (d, J = 7.35 Hz, 3H).

[0095] Example 10: Synthesis of compound B-12

[0096]

[0097] Compound B-12 was characterized by structural parameters: 1 H NMR (400 MHz, DMSO-d6) δ 12.68 (s, 1H), 9.31 (t, J = 5.86 Hz, 3H), 8.26 (s, 1H), 8.00 (d, J = 8.57 Hz, 1H), 7.51 (d, J = 7.53 Hz, 1H), 7.44 (t, J = 7.88 Hz, 1H), 3.94 (d, J = 5.94 Hz, 4H).

[0098] Example 11: Synthesis of compound B-13

[0099]

[0100] Compound B-13 was characterized by structural parameters: 1 H NMR (400 MHz, DMSO-d6) δ 12.64 (s, 1H), 9.15 (d, J = 7.25 Hz, 1H), 8.35 (s, 1H), 7.99 (d, J = 8.84 Hz, 1H), 7.51 (d, J = 7.70 Hz, 1H), 7.49-7.38 (m, 1H), 4.45-4.32 (m, 1H), 1.39 (d, J = 7.35 Hz, 3H).

[0101] Example 12: Synthesis of compound C-4

[0102]

[0103] Step 1: Dissolve 5-bromo-2-fluorobenzaldehyde (0.29 ml, 2.46 mmol, 1.0 eq) in 5 ml DMF, add ethyl mercaptoacetate (0.41 ml, 3.69 mmol, 1.5 eq), potassium carbonate (1.02 g, 7.39 mmol, 3 eq), and warm to 75 °C. Stir the reaction mixture at 75 °C for 1.5-3 h. Monitor the reaction by TLC (V petroleum ether:V ethyl acetate = 40:1). When the starting material is no longer reduced, stop the reaction. After the reaction is completed, concentrate the DMF, extract the reaction system with EA (3 x 30 ml), wash the organic phase with H2O (2 x 30 ml) successively, wash with saturated brine (1 x 30 ml), combine the organic phases, dry over anhydrous Na2SO4, filter, and purify the crude product obtained after concentration under reduced pressure by silica gel column chromatography (PE:EA = 80:1) to obtain white solid, which is compound 23b (646 mg, 91%). 1 H NMR (400 MHz, DMSO-d6) δ 10.35 (s, 1H), 9.75 (s, 1H), 8.30 (s, 1H), 8.05-7.92 (m, 2H), 7.60 (d, J = 8.5 Hz, 1H), 7.51-7.38 (m, 2H), 6.98-6.83 (m, 2H).

[0104] Example 13: Synthesis of compound D-1

[0105]

[0106] Step 1: Dissolve 5-bromo-2-fluorobenzaldehyde (0.29 ml, 2.46 mmol, 1.0 eq) in 5 ml DMF, add ethyl mercaptoacetate (0.41 ml, 3.69 mmol, 1.5 eq), potassium carbonate (1.02 g, 7.39 mmol, 3 eq), and warm to 75 °C. Stir the reaction mixture at 75 °C for 1.5-3 h. Monitor the reaction by TLC (V petroleum ether:V ethyl acetate = 40:1). When the starting material is no longer reduced, stop the reaction. After the reaction is completed, concentrate the DMF, extract the reaction system with EA (3 x 30 ml), wash the organic phase with H2O (2 x 30 ml) successively, wash with saturated brine (1 x 30 ml), combine the organic phases, dry over anhydrous Na2SO4, filter, and purify the crude product obtained after concentration under reduced pressure by silica gel column chromatography (PE:EA = 80:1) to obtain white solid, which is compound 23b (646 mg, 91%). 1H NMR (400 MHz, Chloroform-d) δ 8.01 (d, J = 1.9 Hz, 1H), 7.96 (d, J = 1.0 Hz, 1H), 7.72 (dt, J = 8.7, 0.7 Hz, 1H), 7.54 (dd, J = 8.7, 1.9 Hz, 1H), 4.41 (q, J = 7.1 Hz, 2H), 1.42 (t, J = 7.1 Hz, 3H).

[0107]

[0108] Step 2: Compound 23b (0.9 g, 3.15 mmol, 1 eq) and Pd(dppf)Cl2(0.51 g, 0.69 mmol, 0.22 eq) were dissolved in a mixture of dioxane / water (V / V = 5 / 1) and subjected to Suzuki coupling reaction, and then the reaction mixture was heated to 80 °C. After 30 min, 3,4-dimethoxynaphthalene boronic acid (0.63 g, 3.47 mmol, 1.1 eq) and Cs2CO3(2.06 g, 6.31 mmol, 2 eq) were added. Then the reaction mixture was stirred at 80 °C for 3.5-4 h. The reaction process was monitored by TLC (V petroleum ether:V ethyl acetate = 10:1), and the reaction was stopped when the starting material was no longer reduced. After the reaction was completed, the reaction system was extracted with EA (3 x 30 ml), and the organic phase was washed successively with H2O (2 x 30 ml), saturated brine (1 x 30 ml), and then combined and dried over anhydrous Na2SO4. After filtration, the concentrated crude product was purified by silica gel column chromatography (PE:EA = 20:1) to obtain white solid, which was compound 23c (787 mg, 73%). 1 H NMR (400 MHz, Chloroform-d) δ 8.08 (d, J = 0.8 Hz, 1H), 8.00 (dd, J = 1.8, 0.6 Hz, 1H), 7.88 (dt, J = 8.5, 0.8 Hz, 1H), 7.65 (dd, J = 8.5, 1.8 Hz, 1H), 7.21 - 7.12 (m, 2H), 6.97 (d, J = 8.3 Hz, 1H), 4.41 (q, J = 7.1 Hz, 2H), 3.95 (d, J = 14.2 Hz, 6H), 1.42 (t, J = 7.1 Hz, 3H).

[0109]

[0110] Step 3: Compound 23c (180 mg, 5.26 mmol, 1 eq) was added to 3 ml MeOH and 3 ml H2O system stirring dissolved, then added NaOH (53 mg, 13.1 mmol, 2.5 eq); the reaction mixture was heated at 60°C for 2-2.5 hours. TLC (V dichloromethane:V methanol = 20:1) monitoring reaction process, when the raw material no longer reduced, the reaction was stopped. After the reaction was completed, the reaction liquid was concentrated, the reaction mixture was extracted with EA (1x30 ml) with the reaction system, washed with saturated NaHCO3 aqueous solution (3x30 ml), the combined aqueous phase was adjusted to PH 2-3 with 10% HCl, and the white solid was precipitated after standing overnight. After filtration, the solid was dried to obtain compound D-1 (112 mg, 67%). 1 H NMR (400 MHz, DMSO-d6) δ 8.13 (d, J = 1.8 Hz, 1H), 7.96 (d, J = 8.5 Hz, 1H), 7.84 (s, 1H), 7.68 (dd, J = 8.5, 1.8 Hz, 1H), 7.28 (d, J = 2.2 Hz, 1H), 7.24 (dd, J = 8.3, 2.2 Hz, 1H), 7.04 (d, J = 8.4 Hz, 1H), 3.86 (s, 3H), 3.80 (s, 3H).

[0111] The synthesis of compounds D-2 to D-3 in the following examples 24 to 25 can refer to the synthesis method of example 23, only the corresponding raw materials need to be changed.

[0112] Example 14: Synthesis of compound D-3

[0113]

[0114] Compound D-3 structure parameter characterization: 1 H NMR (400 MHz, DMSO-d6) δ 8.13 (d, J = 1.8 Hz, 1H), 7.96 (d, J = 8.5 Hz, 1H), 7.84 (s, 1H), 7.68 (dd, J = 8.5, 1.8 Hz, 1H), 7.28 (d, J = 2.2 Hz, 1H), 7.24 (dd, J = 8.3, 2.2 Hz, 1H), 7.04 (d, J = 8.4 Hz, 1H), 3.86 (s, 3H), 3.80 (s, 3H).

[0115] Example 15: Synthesis of compound D-4

[0116]

[0117]

[0118] Step 1 : Dissolve 4-bromo-2-hydroxybenzaldehyde (0.29 ml, 2.46 mmol, 1.0 eq) in 5 ml DMF, add methyl bromoacetate (0.41 ml, 3.69 mmol, 1.5 eq), potassium carbonate (1.02 g, 7.39 mmol, 3 eq), and warm to 130 °C. Stir the reaction mixture at 130 °C for 4-5 h. Monitor the reaction by TLC (V petroleum ether:V ethyl acetate = 40:1). When the starting material is no longer consumed, stop the reaction. After the reaction is complete, concentrate the DMF, and extract the reaction with EA (3 x 30 ml). Wash the organic phase with H2O (2 x 30 ml), saturated brine (1 x 30 ml), combine the organic phases, dry over anhydrous Na2S04, filter, and concentrate the crude product under reduced pressure. Purify the product by silica gel column chromatography (PE:EA = 70:1) to obtain compound 26b (646 mg, 91%) as a white solid. 1 HNMR (400 MHz, Chloroform-d) δ 7.74 (dt, J = 1.6, 0.7 Hz, 1H), 7.53 (dd, J = 8.5, 0.5 Hz, 1H), 7.47 (d, J = 1.0 Hz, 1H), 7.42 (dd, J = 8.4, 1.6 Hz, 1H), 3.96 (s, 3H).

[0119] Step 2: Refer to the method of Example 23, replace 23a with 26a to prepare compound D-4. 1 H NMR (400 MHz, DMSO-d6) δ 7.95 (dt, J = 1.6, 0.8 Hz, 1H), 7.77 (d, J = 8.3 Hz, 1H), 7.70 - 7.58 (m, 2H), 7.31 - 7.22 (m, 2H), 7.01 (s, 1H), 3.83 (s, 3H), 3.77 (s, 3H).

[0120] Example 16: Synthesis of compound F-2

[0121]

[0122] Compound F-2 structure parameter characterization: 1H NMR (400 MHz, DMSO-d6) δ 9.03 (d, J = 7.3 Hz, 1H), 8.24 (s, 1H), 8.20 (d, J = 1.8 Hz, 1H), 8.07 (d, J = 8.5 Hz, 1H), 7.77 (dd, J = 8.5, 1.9 Hz, 1H), 7.34 - 7.26 (m, 2H), 7.07 (dd, J = 8.4, 0.9 Hz, 1H), 4.44 (td, J = 7.4, 1.3 Hz, 1H), 3.85 (d, J = 26.9 Hz, 6H), 1.44 (d, J = 7.4 Hz, 3H).

[0123] Example 17: Synthesis of compound F-3

[0124]

[0125] Compound F-3 was characterized by structural parameters: 1 H NMR (400 MHz, Chloroform-d) δ 7.94 (s, 1H), 7.85 (d, J = 8.5 Hz, 1H), 7.75 (s, 1H), 7.63 (d, J = 8.4 Hz, 1H), 7.35 - 7.19 (m, 5H), 7.15 (dd, J = 8.3, 1.9 Hz, 1H), 7.12 (d, J = 2.0 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 6.61 (d, J = 6.9 Hz, 1H), 5.11 (s, 1H), 3.93 (d, J = 11.3 Hz, 6H), 3.37 (dd, J = 13.8, 5.2 Hz, 1H), 3.33 - 3.25 (m, 1H).

[0126] Example 18: Synthesis of compound F-4

[0127]

[0128] Compound F-4 was characterized by structural parameters: 1 H NMR (400 MHz, DMSO-d6) δ 9.19 (t, J = 5.9 Hz, 1H), 8.27 - 8.14 (m, 2H), 8.11 (d, J = 8.5 Hz, 1H), 7.85 - 7.67 (m, 3H), 7.50 (t, J = 7.6 Hz, 2H), 7.39 (t, J = 7.3 Hz, 1H), 3.97 (d, J = 5.9 Hz, 2H).

[0129] Example 19: Synthesis of compound F-7

[0130]

[0131] Compound F-7 structure parameter characterization: 1 H NMR (400 MHz, DMSO-d6) δ 9.21 (t, J = 5.9 Hz, 1H), 8.26 (d, J = 1.8 Hz, 1H), 8.20 - 8.10 (m, 2H), 7.93 - 7.86 (m, 2H), 7.78 (dd, J = 8.5, 1.9 Hz, 1H), 7.52 - 7.45 (m, 2H), 3.96 (d, J = 5.8 Hz, 2H).

[0132] Example 20: Synthesis of compound F-8

[0133]

[0134] Compound F-8 structure parameter characterization: 1 H NMR (400 MHz, DMSO-d6) δ 9.15 (d, J = 8.2 Hz, 1H), 8.30 - 8.20 (m, 2H), 8.10 (d, J = 8.5 Hz, 1H), 7.92 - 7.85 (m, 2H), 7.77 (dd, J = 8.5, 1.8 Hz, 1H), 7.53 - 7.45 (m, 2H), 7.36 - 7.31 (m, 2H), 7.27 (t, J = 7.5 Hz, 2H), 7.21 - 7.15 (m, 1H), 4.63 (ddd, J = 10.6, 8.1, 4.4 Hz, 1H), 3.09 (dd, J = 13.8, 10.6 Hz, 2H).

[0135] Example 21: Synthesis of compound F-11

[0136]

[0137] Compound F-11 structure parameter characterization: 1 H NMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 7.4 Hz, 1H), 7.93 (s, 1H), 7.78 (d, J = 8.2 Hz, 1H), 7.63 (dd, J = 8.2, 1.5 Hz, 1H), 7.54 (s, 1H), 7.38 - 7.17 (m, 5H), 7.17 - 7.10 (m, 1H), 7.04 (d, J = 8.3 Hz, 1H), 4.51 (d, J = 6.5 Hz, 1H), 3.82 (d, J = 24.6 Hz, 6H), 3.13 (dd, J = 13.7, 8.2 Hz, 2H).

[0138] Example 22: Synthesis of compound F-14

[0139]

[0140] Compound F-14 structure parameter characterization: 1 H NMR (400 MHz, DMSO-d6) δ 11.62 (d, J = 2.2 Hz, 1H), 8.86 (t, J = 6.0 Hz, 1H), 7.87 (s, 1H), 7.54 - 7.37 (m, 2H), 7.23 (d, J = 2.1 Hz, 1H), 7.20 - 7.13 (m, 2H), 7.01 (d, J = 8.5 Hz, 1H), 3.97 (d, J = 5.9 Hz, 2H), 3.82 (d, J = 29.0 Hz, 6H).

[0141] Example 23: Pharmacological experiments

[0142] Cell culture and ISRE-luciferase assay

[0143] Wild type (cat. no. thpl-isg), cGAS KO (cat. no. thpd-kocgas) and STING KO (cat. no. thpd-kostg) ISG-THP-1 cells were purchased from Invivogen. The inhibitor SN-011 was used as a positive control compound.

[0144] Cells were cultured in RPMI 1640 (Biological Industries) containing 10% heat-inactivated fetal bovine serum (FBS, Biological Industries), 1% (v / v) penicillin-streptomycin (Biological Industries), 0.25 pg / mL amphotericin B (Invitrogen) and 100 pg / mL bovine albumin (Invitrogen) at 37°C in a 5% CO2 incubator.

[0145] Cells were resuspended in low serum medium (2% fetal bovine serum) at a concentration of 5 x 105 cells / ml and seeded in 96-well white Greiner plates. After 12 h of treatment with different concentrations of compounds or vehicle (DMSO), cells were incubated with 50 pL Quanti-luc (Invivogen) detection reagent. Luciferase activity was measured using a 96-well plate reader (BioTek).

[0146] For each cell type, the luminescence signal of the test article sample was normalized to the vehicle-treated sample and reported as relative light units (RLU).

[0147] Negative values indicate inhibition, and the smaller the value, the better the activity.

[0148] Table 1. Results of ISRE-luciferase assay with SN-011 as a positive control compound for some of the compounds

[0149]

[0150]

[0151] The RLU values of the compounds shown in Table 1 (except F-14) are significantly lower than the positive control compound SN-011, indicating that the compounds in this part have certain inhibitory activity on the cGAS-STING signaling pathway and can be used for the treatment of autoimmune diseases. It can be seen that the compounds have good regulation effect on STING, and have significant advantages and further development potential compared with the STING regulators reported so far.

[0152] The above examples are only used to illustrate the technical solutions of the present application and not to limit it, and the specific embodiments of the present application can be modified or some technical features can be replaced by equivalents; all of which should be covered in the scope of the technical solutions claimed by the present application.

Claims

1. A pharmaceutical compound targeting STING, characterized in that The compound is any one of the benzoheterocyclic compounds or pharmaceutically acceptable salts of the following structural formula: A-1： B-3： B-4： B-7： B-8： B-9： B-10： B-11： B-12： C-4： D-1： D-3： D-4： F-2: F-3: F-4: F-7: F-8： F-11： F-14： 2. The pharmaceutical compound targeting STING according to claim 1, characterized in that When the benzoheterocyclic compound is used as a pharmaceutically acceptable salt, the salt is an acid salt of at least one of the following acids: benzenesulfonic acid, citric acid, glycolic acid, lactic acid, malic acid, malonic acid, mandelic acid, phosphoric acid, hydrochloric acid, sulfuric acid, tartaric acid, succinic acid, formic acid, hydroiodic acid, hydrobromic acid, methanesulfonic acid, nicotinic acid, nitric acid, orotic acid, oxalic acid, pyroglutamic acid, saccharinic acid, salicylic acid, gentisic acid, p-toluenesulfonic acid, valeric acid, palmitic acid, stearic acid, carbonic acid, ethanedisulfonic acid, ethylsuccinic acid and fumaric acid.

3. The method for preparing a pharmaceutical compound targeting STING according to claim 1, wherein The preparation reaction formula of compound B-3 is as follows: The R1 substituent group on the general formula d corresponds to the substituent group on the B-3 compound.

4. The method for preparing a pharmaceutical compound targeting STING according to claim 1, wherein The preparation reaction formula of D-3 or F-4 or F-7 or F-8 compound is as follows: wherein the R1 and R2 substituent groups on the general formula e correspond to the substituent groups on the F-4 or F-7 or F-8 compounds, respectively; The R1 substituent groups on Formula C correspond to the substituent groups on the D-3 compound.

Images