CN113248475B - Heterocyclic substituted benzimidazole dimer or pharmaceutically acceptable salt, composition and application thereof - Google Patents
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Abstract
Heterocyclic substituted benzimidazole dimers represented by the following general formula I or pharmaceutically acceptable salts, compositions and application thereof. The heterocycle-substituted benzimidazole dimer has the activity of activating interferon gene stimulating protein, has improved water solubility, and can be used as a potential medicament for treating tumors or infectious diseases.
Description
Technical Field
The invention belongs to the field of medicinal chemistry, and particularly relates to a heterocyclic substituted benzimidazole dimer or pharmaceutically acceptable salt thereof, a composition and application thereof.
Background
Innate immunity is the first line of defense of the body against pathogen infection, and plays a crucial role in inhibiting tumor growth and the pathogenesis of autoimmunity. In recent years, cGAS-STING-TBK1 pathway has attracted much attention as natural immune regulation, and when cGAS senses pathogen DNA, the DNA receptor cyclic guanosine monophosphate synthase (cGAS) induces cyclic guanosine monophosphate (cGAMP) to activate Interferon gene stimulating factors (stimulants of Interferon genes, STING), recruits TANK binding kinase 1 (TANK-binding kinase 1, TBK 1) to phosphorylate and activate Interferon regulatory Factor 3 (Interferon regulatory Factor 3, irf 3), induces production of type I interferons and cytokines, and activates adaptive immune system through a series of cascade reactions to activate T cells to exert antitumor immune effect.
The STING agonist not only induces the expression of type I interferon genes, plays an important role in natural immune signal pathways, but also activates immunostimulatory cells including dendritic cells and the like, changes the tumor microenvironment and induces the generation of tumor specific T cells, thereby killing tumor cells. Research has shown that: primary tumor growth and distant lesions were prevented by intratumoral or intravenous injection of STING agonists in different mouse tumor models, including B16 melanoma model, 4T1 breast cancer and CT26 colon cancer model. These findings have become important strategies for anti-tumor immunotherapy by activating STING.
To date, the study of STING small molecule agonists is still in the initiative and only four compounds are in clinical trial studies, with ADU-S100 (phase II) developed by the adoro company and MK1454 (phase I) developed by the merck company, both cyclic dinucleotide analogues and the mode of administration being intratumoral injection. The drugs have the defects of large molecular weight, poor cell membrane permeability, negative phosphate group contained in the structure, very easy hydrolysis of phosphate bonds, very poor PK property and the like, and the clinical use of the compounds is severely limited. The other two compounds do not disclose structures. In 2018, the GSK company reports the first benzimidazole STING agonist capable of being administrated by intravenous injection (Nature 2018,564, 439-443.), but the compounds still have serious problems of solubility and metabolism.
Disclosure of Invention
The technical object of the present invention is to provide a compound having high water solubility and good STING agonistic activity, a composition comprising the same and use thereof.
In one aspect, the present invention provides a class of heterocyclic substituted benzimidazole dimers represented by the following general formula I:
in the above general formula I, the compound of formula I,
T 1 、T 2 each independently selected from cyano, a 5-6 membered heteroaromatic ring containing nitrogen heteroatoms, a 4-6 membered heterocyclic ring, carbonyl, provided that T 1 And T 2 Not being carbonyl at the same time;
when T is 1 Or T 2 When it is cyano, R 1 、R 2 Is absent; when T is 1 Or T 2 When not cyano, R 1 、R 2 Each independently selected from H, substituted or unsubstituted C1-C3 alkyl, amino, hydroxy, halogen, substituted with amino, hydroxy or halogen;
R 3 and R 4 Each independently selected from H, substituted or unsubstituted C1-C3 alkoxy, said substitution being by amino, hydroxy, halogen, cyano or a 4-6 membered heterocyclic ring;
R 5 、R 6 each independently selected from substituted or unsubstituted C1-C3 alkyl, said substitution being by hydroxy, halogen or cyano.
In one embodiment, R 1 -T 1 -、R 2 -T 2 -each independently is cyano, triazolyl, imidazolyl, 4,5-dihydrooxazolyl or-C (= O) NH 2 Provided that R is 1 -T 1 -and R 2 -T 2 -not simultaneously being-C (= O) NH 2 ；
R 3 Selected from H or C1-C3 alkoxy, R 4 Selected from C1-C3 alkoxy substituted by morpholinyl or hydroxy;
R 5 、R 6 each independently is C1-C3 alkyl which is unsubstituted or substituted by halogen.
In one embodiment, R 1 -T 1 -、R 2 -T 2 -each independently is cyano, 1,2,4-triazolyl, imidazolyl, 4,5-dihydrooxazolyl or-C (= O) NH 2 Provided that R is 1 -T 1 -and R 2 -T 2 -not simultaneously being-C (= O) NH 2 ；
R 3 Selected from H or methoxy, R 4 Selected from propoxy substituted with morpholinyl or hydroxy;
R 5 、R 6 each independently is ethyl unsubstituted or substituted with halogen, preferably the halogen is F.
In one embodiment, the heterocyclic substituted benzimidazole dimers represented by formula I may be selected from one of the following compounds:
in particular embodiments, the heterocycle-substituted benzimidazole dimers represented by formula I may also include zwitterionic or isomeric forms thereof.
In particular embodiments, the pharmaceutically acceptable salts can include salts of heterocyclic substituted benzimidazole dimers represented by formula I with the following acids: inorganic acids such as phosphoric acid, sulfuric acid, and hydrochloric acid, organic acids such as acetic acid, tartaric acid, citric acid, and malic acid, or acidic amino acids such as aspartic acid and glutamic acid.
In another aspect, the present invention provides a pharmaceutical composition comprising the heterocyclic substituted benzimidazole dimer represented by the general formula I or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.
In another aspect, the invention provides the use of the heterocyclic substituted benzimidazole dimer represented by the general formula I or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition in the preparation of an interferon gene stimulating factor agonist.
In particular embodiments, the interferon gene stimulating factor agonists are useful for treating tumors or infectious diseases.
In particular embodiments, the tumor may be selected from: brain and spinal cancers, head and neck cancers, leukemias and blood cancers, skin cancers, cancers of the reproductive system, cancers of the gastrointestinal system, esophageal cancers, nasopharyngeal cancers, pancreatic cancers, rectal cancers, hepatocellular cancers, cholangiocarcinomas, gallbladder cancers, colon cancers, multiple myeloma, kidney and bladder cancers, bone cancers, lung cancers, malignant mesothelioma, sarcomas, lymphomas, adenocarcinoma, thyroid cancers, cardiac tumors, germ cell tumors, malignant neuroendocrine tumors, malignant rhabdoid tumors, soft tissue sarcomas, midline tract cancers, and cancers of unknown primary origin.
In particular embodiments, the infectious disease may be selected from: viral infectious diseases, such as human immunodeficiency virus, herpes simplex virus, hepatitis B virus, hepatitis C virus; pathogenic microorganism infectious diseases.
In still another aspect, the present invention provides a heterocyclic substituted benzimidazole dimer represented by the general formula I or a pharmaceutically acceptable salt thereof, or a use of the pharmaceutical composition in preparing an immunological composition or a vaccine adjuvant.
Advantageous effects
The novel benzimidazole dimer prepared by the invention has the activity of activating the human interferon gene stimulating protein in THP1 cells, so that the compounds have great potential in the treatment of tumors and infectious diseases.
In addition, various heterocycles and other substituent groups are introduced into the benzimidazole dimer, so that the water solubility is improved while the high-activation activity of the benzimidazole dimer on STING is maintained. The invention proves through experiments that the water solubility of the compound S1 hydrochloride in the application reaches 71.33mg/mL, which is more than 20 times of the hydrochloride of the compound 3 (hereinafter referred to as the compound 1A) in the Nature 2018,564,439-443, and the compound S1 in the application has the stimulation protein multiple of the human interferon gene in the THP1-Blue-ISG cells of the human source of 19.21, which is slightly stronger than the compound 1A (16.93).
Detailed Description
Preparation examples
The following preparation examples exemplarily show specific preparation methods of some compounds of general formula I (S1 to S21) of the present invention. Synthesis of Compound 1A is described in Nature 2018,564,439-443. However, the following examples are only for the purpose of making the present invention easier to understand, and do not limit the scope of the present invention.
Preparative example 1 synthesis of compound S1:
synthesis of Compounds 1-2:
compound 1-1 (2g, 5.28mmol) was dissolved in N, N-dimethylformamide dimethyl acetal (DMF-DMA) (10 ml), and stirred at 100 ℃ for 1 hour. After the reaction is completed, cooling to room temperature, and spin-drying to obtain the compound 1-2.[ Synthesis of Compound 1-1 reference CN109071514A ]
Synthesis of Compounds 1-3:
compound 1-2 (500mg, 1.15mmol) was dissolved in dichloromethane (10 ml) at 0 ℃ and glacial acetic acid (0.26 ml) and hydrazine hydrate (0.11ml, 2.23mmol) were added. The mixture was heated and stirred at 40 ℃ for 0.5 hour. After the reaction is completed, cooling to room temperature, carrying out spin drying, pulping with water, filtering, carrying out press drying on a filter cake, pulping with a small amount of ethanol, filtering again, and drying the solid to obtain the compound 1-3.
Synthesis of Compounds 1-4:
compound 1-3 (425mg, 1.05mmol) was dissolved in anhydrous methanol (3 ml), and a solution of 4M hydrogen chloride in 1,4-dioxane (3 ml) was added, followed by stirring at room temperature for 1 hour. After the reaction is completed, spin-drying, adding methanol for dissolving, and then spin-drying. Repeating twice, finally adding methanol (2 ml) for pulping, filtering out solid, and drying to obtain the compound 1-4.
Synthesis of Compounds 1-6:
the compound 1-4 (325mg, 0.95mmol) was weighed in a sealed tube, isopropanol (2 ml) was added thereto, and N, N-Diisopropylethylamine (DIPEA) (0.05 ml) was added dropwise with stirring, followed by stirring at room temperature for 2 minutes. Then, compound 1-5 (265mg, 0.77mmol) and N, N-diisopropylethylamine (0.13ml, 7.7mmol) were added. The reaction was heated to 120 ℃ for 48 hours. After the reaction is completed, the reaction solution is directly mixed with silica gel and loaded on a column, and the compound 1-6 is obtained after column purification. [ Synthesis of Compounds 1 to 5 refer to Nature 2018,564,439-443]
Synthesis of Compounds 1-7:
compound 1-6 (103mg, 0.17mmol) was dissolved in tetrahydrofuran (2 ml) and water (2 ml), and sodium dithionite (293mg, 1.70mmol) was added and stirred at room temperature for 10 minutes under nitrogen. After the reaction is completed, directly stirring a sample with silica gel, loading the sample on a column, and purifying the sample through the column to obtain the compounds 1 to 7.
Synthesis of compound S1:
compounds 1-7 (48mg, 0.087mmol) were dissolved in N, N-Dimethylformamide (DMF) (0.8 ml) and protected with nitrogen. A solution of 0.4M compound 1-8 in 1,4-dioxane (0.44ml, 0.17mmol) was added dropwise at 0 deg.C. After stirring at 0 ℃ for 1 hour, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (42mg, 0.22mmol) was added, followed by dropwise addition of triethylamine (0.05ml, 0.52mmol), and the reaction was allowed to warm to room temperature for 12 hours. After the reaction is completed, the reaction solution is directly mixed with a sample and loaded on a column, and the compound S1 is obtained by column chromatography purification.
1 H NMR (400 MHz, methanol-d) 4 ) δ 8.39 (s, 1H), 7.62 (d, J =1.2hz, 1h), 7.47 (d, J =1.3hz, 1h), 7.26 (d, J =1.3hz, 1h), 7.08 (d, J =1.4hz, 1h), 6.54 (s, 1H), 6.46 (s, 1H), 5.72-5.58 (m 2H), 4.87-4.82 (m, 4H), 4.58-4.47 (m, 4H), 3.78 (t, J =6.2hz, 2h), 3.63 (s, 3H), 3.54-3.48 (m, 4H), 2.28 (d, J =7.4hz, 2h), 2.27-2.19 (m, 4H), 2.16 (s, 3H), 2.15 (s, 3H), 1.73-1.68 (m, 1.34H), 1.6 (m, 3H). [ Synthesis of Compounds 1-8 reference CN109071514A]
Preparative example 2 synthesis of compound S2:
synthesis of Compound 2-2:
the synthesis of compound 2-2 was identical to that of compound 1-2, except that compound 2-1 was used instead of compound 1-1. [ Synthesis of Compound 2-1 reference CN109071514A ]
Synthesis of Compounds 2-3:
the synthesis of compound 2-3 was the same as the synthesis of compound 1-3, except that compound 2-2 was used instead of compound 1-2.
Synthesis of Compounds 2-4:
the synthesis of compounds 2-4 was identical to that of compounds 1-7, except that compounds 2-3 were used instead of compounds 1-6.
Synthesis of compound S2:
the synthesis of compound S2 was identical to that of compound S1, except that compounds 2-4 were used instead of compounds 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ8.25(s,2H),7.48((d,J＝2.4Hz,2H),7.03(d,J＝2.6Hz,2H),6.43(d,J＝2.7Hz,2H),5.55–5.41(m,2H),4.67–4.58(m,4H),4.45–4.25(m,4H),3.65(t,J＝5.5Hz,2H),3.61(s,3H),3.58–3.49(m,4H),2.51–2.47(m,6H),2.10(s,3H),2.06(s,3H),1.77–1.72(m,2H),1.22–1.19(m,6H)。
Preparative example 3. Synthesis of compound S3:
synthesis of Compound 3-1:
compounds 1-5 (1g, 2.9 mmol) and phosphorus oxychloride (10 ml) were mixed and heated at 100 ℃ overnight. After the reaction was completed, the reaction solution was spin-dried, and ice water (100 ml) was added thereto and stirred uniformly. Then, the mixture is filtered, and the solid is dried to obtain a compound 3-1.
Synthesis of Compound 3-2:
the synthesis of compound 3-2 was identical to that of compound 1-6, except that compound 3-1 was used instead of compound 1-5.
Synthesis of Compounds 3-3:
the synthesis of compound 3-3 was identical to that of compound 1-7, except that compound 3-2 was used instead of compound 1-6.
Synthesis of compound S3:
the synthesis of compound S3 was identical to that of compound S1, except that compound 3-3 was substituted for compound 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ8.38(s,1H),7.51(s,1H),7.14(s,1H),7.08(s,1H),6.64(s,1H),6.45–6.34(m,2H),5.65–5.51(m,2H),4.77–4.63(m,4H),4.49–4.44(m,4H),3.66(t,J＝6.2Hz,2H),3.58(s,3H),3.49–3.34(m,4H),2.24(s,2H),2.21(d,J＝6.7Hz,4H),2.10(s,6H),1.59–1.52(m,2H),1.28–1.21(m,6H)。
Preparative example 4 synthesis of compound S4:
synthesis of Compound 4-2:
the synthesis of compound 4-2 was identical to the synthesis of compound 3-2, except that compound 4-1 was used instead of compound 1-4. [ Synthesis of Compound 4-1 reference CN109071514A ]
Synthesis of Compounds 4-3:
the synthesis of compound 4-3 was identical to that of compound 1-7, except that compound 4-2 was used instead of compound 1-6.
Synthesis of compound S4:
the synthesis of compound S4 was identical to that of compound S1, except that compound 4-3 was used instead of compound 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ7.38(s,1H),7.24(d,J＝1.7Hz,1H),7.24(t,J＝0.8Hz,1H),6.88(d,J＝5.6Hz,1H),6.58–6.52(m,2H),5.80(s,2H),4.93–4.86(m,4H),4.59–4.52(m,4H),3.95–3.91(m,2H),3.72(s,3H),3.62–3.54(m,4H),2.30(s,6H),2.21–2.15(m,6H),1.76–1.71(m,2H),1.39–1.32(m,6H)。
Preparative example 5. Synthesis of compound S5:
synthesis of Compound 5-2:
compound 5-1 (1g, 4.35mmol) was mixed with phosphorus oxychloride (10 ml), and the mixture was stirred at 100 ℃ for 12 hours. After the reaction is completed, cooling to room temperature, spin-drying the reaction solution, adding ice water (100 ml), stirring at room temperature to uniformly suspend the solid in water, performing suction filtration, and drying the solid to obtain the compound 5-2.[ Synthesis of Compound 5-1 reference CN109071514A ]
Synthesis of Compounds 5-4:
compound 5-2 (674mg, 3.18mmol) was dissolved in anhydrous methanol (10 ml), and a 5.4M methanol solution (0.3ml, 1.6mmol) of sodium methoxide was slowly added dropwise thereto, followed by stirring at room temperature for 3 hours under nitrogen protection. Then, compound 5-3 (0.35ml, 3.18mmol) and glacial acetic acid (0.4ml, 6.36mmol) were added, and the mixture was heated to 50 ℃ under nitrogen and stirred for 2 hours. After this time, 6N hydrochloric acid was added dropwise to the reaction solution to adjust pH =1, and the mixture was heated to 70 ℃ and refluxed for 18 hours. After the reaction was complete, it was cooled to room temperature. Spin-dry with 2M aqueous sodium carbonate solution to give a mixture pH = 10. Then filtering, washing the solid with petroleum ether to obtain the compound 5-4.
Synthesis of Compounds 5-6:
compound 5-4 (330mg, 1.3mmol), compound 5-5 (291mg, 1.56mmol) and N, N-diisopropylethylamine (0.68ml, 4 mmol) were suspended in ethanol (5 ml), placed in a sealed tube, and stirred at 120 ℃ for 24 hours. After the reaction is completed, cooling to room temperature, directly stirring the reaction solution to a column, and purifying by the column to obtain the compound 5-6.
Synthesis of Compounds 5-7:
the synthesis of compounds 5-7 was identical to that of compounds 1-4, except that compounds 5-6 were used instead of compounds 1-3.
Synthesis of Compounds 5-8:
the synthesis of compounds 5-8 was identical to that of compounds 1-6, except that compounds 5-7 were used instead of compounds 1-4.
Synthesis of Compounds 5 to 9
The synthesis of compounds 5-9 was identical to that of compounds 1-7, except that compounds 5-8 were used instead of compounds 1-6.
Synthesis of compound S5:
the synthesis of compound S5 was identical to that of compound S1, except that compounds 5-9 were used instead of compounds 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ7.41(d,J＝1.1Hz,1H),7.37(d,J＝1.2Hz,1H),7.10(s,2H),7.03(s,1H),7.01(s,1H),6.48(s,1H),6.40(s,1H),5.63–5.51(m,2H),4.77–4.70(m,4H),4.51–4.46(m,2H),4.46–4.41(m,2H),3.68(t,J＝5.9Hz,2H),3.52(s,3H),3.50(d,J＝4.5Hz,4H),2.20–2.13(m,6H),2.11(s,3H),2.10(s,3H),1.56(t,J＝7.6Hz,2H),1.29–1.22(m,6H)。
Preparative example 6 synthesis of compound S6:
synthesis of Compound 6-2:
the synthesis of compound 6-2 was identical to that of compound 1-2, except that compound 6-1 was used instead of compound 1-1. [ Synthesis of Compound 6-1 reference CN109071514A ]
Synthesis of Compounds 6-3:
the synthesis of compound 6-3 was identical to that of compound 1-3, except that compound 6-2 was used instead of compound 1-2.
Synthesis of Compounds 6-4:
the synthesis of compound 6-4 was identical to that of compound 1-7, except that compound 6-3 was used instead of compound 1-6.
Synthesis of compound S6:
the synthesis of compound S6 was identical to that of compound S1, except that compounds 6-4 were used instead of compounds 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ8.29(s,1H),8.26(s,1H),7.87(s,1H),7.59(d,J＝8.4Hz,1H),7.43(s,1H),7.07(s,1H),7.04(d,J＝8.4Hz,1H),6.44(s,1H),6.35(s,1H),5.77–5.56(m,2H),4.71(d,J＝4.9Hz,2H),4.60(d,J＝5.1Hz,2H),4.49–4.35(m,4H),3.73(t,J＝6.4Hz,2H),3.52(t,J＝4.6Hz,4H),2.20–2.11(m,6H),2.09(s,3H),2.02(s,3H),1.58(t,J＝7.5Hz,2H),1.25(t,J＝7.1Hz,3H),1.17(t,J＝7.1Hz,3H)。
Preparative example 7. Synthesis of compound S7:
synthesis of Compound 7-1:
the synthesis of compound 7-1 was identical to that of compound 5-6, except that compound 5-2 was used instead of compound 5-4.
Synthesis of Compound 7-2:
the synthesis of compound 7-2 was identical to that of compound 1-4, except that compound 7-1 was used instead of compound 1-3.
Synthesis of Compounds 7-3:
the synthesis of compound 7-3 was identical to that of compound 3-2, except that compound 7-2 was used instead of compound 1-4.
Synthesis of Compounds 7-4:
the synthesis of compound 7-4 was identical to that of compound 1-7, except that compound 7-3 was used instead of compound 1-6.
Synthesis of compound S7:
the synthesis of compound S7 was identical to that of compound S1, except that compounds 7-4 were used instead of compounds 1-7.
1 H NMR(400MHz,Chloroform-d)δ7.26(s,1H),7.24(s,1H),6.91(s,1H),6.88(s,1H),6.57(s,1H),6.55(s,1H),5.81–5.73(m,2H),4.94–4.82(m,4H),4.61–4.51(m,4H),3.96(s,2H),3.73(s,3H),3.64–3.57(m,4H),2.31–2.21(m,6H),2.20(s,3H),2.20(s,3H),1.78(s,2H),1.41–1.31(m,6H)。
Preparative example 8 synthesis of compound S8:
synthesis of Compound 8-1:
the synthesis of compound 8-1 was identical to that of compound 5-4, except that compound 3-1 was used instead of compound 5-2.
Synthesis of Compound 8-2:
the synthesis of compound 8-2 was identical to that of compound 5-8, except that compound 8-1 was used instead of compound 1-5.
Synthesis of Compounds 8-3:
the synthesis of compound 8-3 was identical to that of compound 1-7, except that compound 8-2 was used instead of compound 1-6.
Synthesis of compound S8:
the synthesis of compound S8 was identical to that of compound S1, except that compound 8-3 was substituted for compound 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ7.28(s,2H),7.09(d,J＝1.2Hz,2H),7.07(d,J＝1.3Hz,2H),7.01–6.97(m,1H),6.97–6.93(m,1H),6.63(s,1H),6.60(s,1H),5.68(d,J＝3.0Hz,2H),4.89–4.81(m,4H),4.60(d,J＝7.0Hz,2H),4.59–4.54(m,2H),3.55–3.42(m,4H),3.49(t,J＝6.5Hz,2H),3.31(s,3H),2.44–2.36(m,4H),2.21(s,3H),2.19(s,3H),2.17(d,J＝7.7Hz,2H),1.51(t,J＝7.4Hz,2H),1.41–1.28(m,6H)。
Preparative example 9 synthesis of compound S9:
synthesis of Compound 9-1:
the synthesis of compound 9-1 was identical to that of compound 1-2, except that compound 1-5 was used instead of compound 1-1.
Synthesis of Compound 9-2：
The synthesis of compound 9-2 was identical to that of compound 1-3, except that compound 9-1 was used instead of compound 1-2.
Synthesis of Compounds 9-3:
the synthesis of compound 9-3 was identical to that of compound 4-2, except that compound 9-2 was used instead of compound 3-1.
Synthesis of Compounds 9-4:
the synthesis of compounds 9-4 was identical to that of compounds 1-7, except that compounds 9-3 were used instead of compounds 1-6.
Synthesis of compound S9:
the synthesis of compound S9 was identical to that of compound S1, except that compounds 9-4 were used instead of compounds 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ8.39(s,1H),7.63(d,J＝1.1Hz,1H),7.55(d,J＝1.2Hz,1H),7.27(s,1H),7.20(d,J＝1.3Hz,1H),6.60(s,1H),6.49(s,1H),5.88–5.71(m,2H),4.94(s,2H),4.60(q,J＝7.1Hz,2H),4.52(q,J＝7.1Hz,2H),3.86(t,J＝6.1Hz,2H),3.68(s,3H),3.60(t,J＝4.6Hz,4H),2.38–2.32(m,2H),2.30(t,J＝5.5Hz,4H),2.21(s,3H),2.13(s,3H),1.74–1.65(m,2H),1.36(t,J＝7.1Hz,3H),1.30(t,J＝7.1Hz,3H)
Preparative example 10 synthesis of compound S10:
synthesis of Compound 10-2:
the synthesis of compound 10-2 was identical to that of compound 1-6, except that compound 10-1 was used instead of compound 1-5.
Synthesis of Compound 10-3:
the synthesis of compound 10-3 was identical to that of compound 1-7, except that compound 10-2 was used instead of compound 1-6.
Synthesis of Compound 10-4:
the synthesis of compound 10-4 was identical to that of compound S1, except that compound 10-3 was used instead of compound 1-7.
Synthesis of compound S10:
compound 10-4 (40mg, 0.044mmol) was dissolved in tetrahydrofuran (1.5 ml) and water (1.5 ml), tetrabutylammonium fluoride (TBFA) (0.087 ml,0.087 mmol) and glacial acetic acid (0.006ml, 0.087 mmol) were added at 0 ℃ and stirred at room temperature for 12 hours. After completion of the reaction, the reaction mixture was spin-dried, and anhydrous methanol (2 ml) was added thereto, followed by stirring at room temperature for 2 hours to precipitate a solid, which was then filtered to obtain Compound S10.
1 H NMR(600MHz,DMSO-d 6 )δ12.81(s,2H),8.47(s,2H),7.96(s,1H),7.81(s,1H),7.63(s,1H),7.46–7.41(m,1H),7.31(s,2H),6.50(d,J＝3.2Hz,2H),5.89–5.80(m,2H),4.92(d,J＝4.1Hz,2H),4.90(d,J＝4.3Hz,2H),4.54–4.48(m,4H),4.06(t,J＝6.4Hz,2H),3.75(s,3H),3.43(t,J＝6.0Hz,2H),2.09(s,3H),2.07(s,3H),1.75–1.68(m,2H),1.28–1.25(m,3H),1.25–1.23(m,3H).
Preparative example 11 synthesis of compound S11:
synthesis of Compound 11-3:
compound 11-1 (770mg, 5 mmol) and cesium carbonate (2.44g, 7.5 mmol) were dissolved in N, N-dimethylformamide (5 ml), and compound 11-2 (943mg, 6.5 mmol) was slowly added thereto under nitrogen protection, and the mixture was stirred at room temperature for 2 hours. After the reaction is completed, the reaction solution is poured into cold water, ethyl acetate is used for extraction for 2 times, and the organic phase is subjected to salt washing and drying. Collecting organic phase silica gel sample, loading the organic phase silica gel sample on a column, and purifying the organic phase silica gel sample through the column to obtain a compound 11-3.
Synthesis of Compounds 11-4:
compound 11-3 (905mg, 4.15mmol) was dissolved in a mixed solution of tetrahydrofuran (5 ml) and water (5 ml), and lithium hydroxide (498mg, 20.75mmol) was added thereto with stirring at room temperature, followed by stirring at room temperature for 1 hour. Then, the tetrahydrofuran is evaporated under reduced pressure, 1N hydrochloric acid is slowly dripped at room temperature until the pH is =6, a white solid is precipitated, the solid is filtered out, and the solid is dried to obtain a compound 11-4.
Synthesis of Compounds 11-5:
compound 11-4 (307mg, 1.62mmol) was weighed out and dichloromethane (5 ml) and N, N-dimethylformamide (0.02 ml) were added under nitrogen. Oxalyl chloride (0.16ml, 1.78mmol) was slowly added dropwise at 0 deg.C, and after addition, stirring was carried out at room temperature for 0.5 hour. After the reaction was complete, the reaction solution was spin dried, dissolved in dichloromethane and spin dried again for 2 times to give a brown solid. Potassium thiocyanate (189mg, 1.95mmol) was dissolved in acetone (5 ml) and purged with nitrogen. The brown solid obtained above was also dissolved in acetone (1 ml) and slowly added dropwise to the reaction solution at 0 ℃. After stirring at 0 ℃ for 5 minutes, potassium thiocyanate (20 mg) was added, and the reaction solution was stirred for 20 minutes while maintaining at 0 ℃. After the reaction was complete, n-hexane (5 ml) was added and the mixture was evaporated to one third of the solvent under reduced pressure. Repeating for 2 times, adding n-hexane (10 ml), stirring, filtering, adding silica gel into the filtrate, mixing with the sample, and loading onto column. Purifying with column to obtain compound 11-5, preparing 0.4M 1,4-dioxane solution of compound 11-5, and storing at-18 deg.C.
Synthesis of compound S11:
the synthesis of compound S11 was identical to that of compound S1, except that compounds 11-5 were used instead of compounds 1-8.
1 H NMR (400 MHz, methanol-d) 4 )δ8.37(s,1H),7.63(d,J＝1.6Hz,1H),7.46(d,J＝1.7Hz,1H),7.23(d,J＝1.7Hz,1H),7.07(d,J＝1.8Hz,1H),6.63(s,1H),6.48(s,1H),5.62–5.54(m,2H),5.23(d,J＝5.5Hz,2H),4.87(s,2H),4.82(s,2H),4.58–4.53(m,2H),4.52–4.45(m,2H),3.77(t,J＝5.8Hz,2H),3.60(s,3H),3.54–3.43(m,4H),2.38(d,J＝7.4Hz,2H),2.36–2.27(m,4H),2.16(s,3H),2.15(s,3H),1.63(t,J＝6.2Hz,2H)。
Preparative example 12 synthesis of compound S12:
synthesis of Compound 12-2:
the synthesis of compound 12-2 was identical to that of compound 11-3, except that compound 12-1 was used instead of compound 11-2.
Synthesis of Compounds 12-3:
the synthesis of compound 12-3 was identical to that of compound 11-4, except that compound 12-2 was used instead of compound 11-3.
Synthesis of Compounds 12-4:
the synthesis of compound 12-4 was identical to that of compound 11-5, except that compound 12-3 was used instead of compound 11-4.
Synthesis of compound S12:
the synthesis of compound S12 was identical to that of compound S1, except that compounds 12-4 were used instead of compounds 1-8.
1 H NMR (400 MHz, methanol-d) 4 )δ8.39(s,1H),7.62(d,J＝1.2Hz,1H),7.47(d,J＝1.3Hz,1H),7.26(d,J＝1.3Hz,1H),7.08(d,J＝1.4Hz,1H),6.54(s,1H),6.46(s,1H),5.72–4.53(m,2H),4.87(s,2H),4.82(s,2H),4.58–4.53(m,4H),4.53–4.47(m,4H),3.78(t,J＝6.2Hz,2H),3.63(s,3H),3.54(t,J＝4.6Hz,4H),2.28(d,J＝7.4Hz,2H),2.26–2.20(m,4H),2.16(s,3H),2.15(s,3H),1.63(t,J＝6.2Hz,2H)。
Preparation example 13 synthesis of compound S13:
synthesis of Compound 13-1:
the synthesis of compound 13-1 was identical to that of compound 1-2, except that compound 10-1 was used instead of compound 1-1.
Synthesis of Compound 13-2:
the synthesis of compound 13-2 was identical to that of compound 1-3, except that compound 13-1 was used instead of compound 1-2.
Synthesis of Compound 13-3:
the synthesis of compound 13-3 was identical to that of compound 4-2, except that compound 13-2 was used instead of compound 3-1.
Synthesis of Compound 13-4:
the synthesis of compound 13-4 was identical to that of compound 1-7, except that compound 13-3 was used instead of compound 1-6.
Synthesis of Compounds 13-5:
the synthesis of compound 13-5 was identical to that of compound S1, except that compound 13-4 was used instead of compound 1-7.
Synthesis of compound S13:
the synthesis of compound S13 was identical to that of compound S10, except that compound 13-5 was used instead of compound 10-4.
1 H NMR (400 MHz, methanol-d) 4 )δ8.26(s,1H),7.62(d,J＝1.2Hz,1H),7.47(d,J＝1.3Hz,1H),7.26(d,J＝1.3Hz,1H),7.08(d,J＝1.4Hz,1H),6.54(s,1H),6.46(s,1H),5.72–5.62(m,2H),4.87(s,2H),4.82(s,2H),4.58–4.47(m,4H),3.78(t,J＝6.2Hz,2H),3.63(s,3H),2.28(d,J＝7.4Hz,2H),2.16(s,3H),2.15(s,3H),1.63(t,J＝6.2Hz,2H),1.34–1.28(m,6H)。
Preparative example 14 synthesis of compound S14:
synthesis of Compound 14-2:
the synthesis of compound 14-2 was identical to that of compound 1-2, except that compound 14-1 was substituted for 1-1. [ Synthesis of Compound 14-1 reference CN109071514A ]
Synthesis of Compound 14-3:
the synthesis of compound 14-3 was identical to that of compound 1-3, except that compound 14-2 was used instead of compound 1-2.
Synthesis of Compound 14-4:
the synthesis of compound 14-4 was identical to that of compound 1-4, except that compound 14-3 was substituted for compound 1-3.
Synthesis of Compounds 14-5:
the synthesis of compound 14-5 was identical to that of compound 1-6, except that compound 14-4 was substituted for compound 1-4.
Synthesis of Compounds 14-6:
the synthesis of compounds 14-6 was identical to that of compounds 1-7, except that compounds 14-5 were used instead of compounds 1-6.
Synthesis of compound S14:
the synthesis of compound S14 was identical to that of compound S11, except that compounds 14-6 were used instead of compounds 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ8.29(s,1H),7.87(s,1H),7.59(d,J＝8.4Hz,1H),7.43(s,1H),7.07(s,1H),7.04(d,J＝8.4Hz,1H),6.44(s,1H),6.35(s,1H),5.77–5.56(m,2H),5.23(d,J＝5.5Hz,2H),4.71(d,J＝4.9Hz,2H),4.60(d,J＝5.1Hz,2H),4.49–4.35(m,4H),3.73(t,J＝6.4Hz,2H),3.52(t,J＝4.6Hz,4H),2.27–2.16(m,6H),2.09(s,3H),2.02(s,3H),1.58(t,J＝7.5Hz,2H)。
Preparative example 15 synthesis of compound S15:
synthesis of Compound 15-1:
the synthesis of compound 15-1 was identical to that of compound 10-2, except that compound 14-4 was used instead of 1-4.
Synthesis of Compound 15-2:
the synthesis of compound 15-2 was identical to that of compound 1-7, except that compound 15-1 was used instead of 1-6.
Synthesis of Compound 15-3:
the synthesis of compound 15-3 was identical to that of compound S11, except that compound 15-2 was used instead of 1-7.
Synthesis of compound S15:
the synthesis of compound S15 was identical to that of compound S10, except that 10-4 was replaced with compound 15-3.
1 H NMR (400 MHz, methanol-d) 4 )δ8.26(s,1H),7.64(d,J＝1.3Hz,1H),7.43(d,J＝1.4Hz,1H),7.21(d,J＝1.6Hz,1H),7.01(d,J＝1.5Hz,1H),6.55(s,1H),6.48(s,1H),5.71–5.63(m,2H),4.84(s,2H),4.71(s,2H),4.53–4.42(m,2H),3.74(t,J＝6.2Hz,2H),3.68(s,3H),2.22(d,J＝7.4Hz,2H),2.19(s,3H),2.17(s,3H),1.76(t,J＝6.8Hz,2H),1.34–1.28(m,6H)。
Preparative example 16 synthesis of compound S16:
synthesis of Compound 16-1:
the synthesis of compound 16-1 was identical to that of compound 10-4, except that compound 12-4 was used instead of 1-8.
Synthesis of compound S16:
the synthesis of compound S16 was identical to that of compound S10, except that 10-4 was replaced with compound 16-1.
1 H NMR (400 MHz, methanol-d) 4 )δ8.39(s,1H),7.62(d,J＝1.2Hz,1H),7.47(d,J＝1.3Hz,1H),7.26(d,J＝1.3Hz,1H),7.08(d,J＝1.4Hz,1H),7.04(d,J＝1.4Hz,1H),6.54(s,1H),6.46(s,1H),5.72(s,2H),4.87(s,2H),4.82(s,2H),4.58–4.47(m,6H),3.78(t,J＝6.2Hz,2H),3.54(t,J＝4.6Hz,2H),2.28(d,J＝7.4Hz,2H),2.16(s,3H),2.15(s,3H),1.63(t,J＝6.2Hz,2H)。
Preparative example 17 synthesis of compound S17:
synthesis of Compound 17-2:
the synthesis of compound 17-1 was the same as that of compound 1-2 except that compound 17-1 was used instead of N, N-dimethylformamide dimethyl acetal.
Synthesis of Compound 17-3:
the synthesis of compound 17-3 was identical to that of compound 1-3, except that compound 17-2 was used instead of 1-2.
Synthesis of Compound 17-4:
the synthesis of compound 17-4 was identical to that of compound 1-4, except that compound 17-3 was substituted for 1-3.
Synthesis of Compounds 17-5:
the synthesis of compound 17-5 was identical to that of compound 1-6, except that compound 17-4 was substituted for 1-4.
Synthesis of Compounds 17-6:
the synthesis of compound 17-6 was identical to that of compound 1-7, except that compound 17-5 was used instead of 1-6.
Synthesis of compound S17:
the synthesis of compound S17 was identical to that of compound S1 except that compounds 17-6 were used instead of 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ7.60(d,J＝1.3Hz,1H),7.51–7.45(m,1H),7.27(d,J＝1.4Hz,1H),7.12(d,J＝1.5Hz,1H),6.58(s,1H),6.49(s,1H),5.75(q,J＝4.4Hz,2H),4.92(d,J＝3.7Hz,2H),4.88(d,J＝3.6Hz,2H),4.58(q,J＝8.5,7.8Hz,2H),4.51(t,J＝7.1Hz,2H),3.83(t,J＝6.1Hz,2H),3.66(s,3H),3.54(t,J＝4.6Hz,4H),2.48(s,3H),2.29(d,J＝7.4Hz,2H),2.26(d,J＝6.4Hz,4H),2.18(s,3H),2.17(s,3H),1.65(t,J＝7.6Hz,2H),1.34(t,J＝7.2Hz,3H),1.30(t,J＝7.1Hz,3H)
Preparative example 18 synthesis of compound S18:
synthesis of compound S18:
the synthesis of compound S18 was identical to that of compound S1, except that compounds 14-6 were used instead of compounds 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ8.29(s,1H),7.87(s,1H),7.59(d,J＝8.4Hz,1H),7.43(s,1H),7.07(s,1H),7.04(d,J＝8.4Hz,1H),6.44(s,1H),6.35(s,1H),5.77–5.56(m,2H),4.71(d,J＝4.9Hz,2H),4.60(d,J＝5.1Hz,2H),4.49–4.35(m,4H),3.73(t,J＝6.4Hz,2H),3.58–3.43(m 4H),2.67–2.54(m,6H),2.09(s,3H),2.02(s,3H),1.58(t,J＝7.5Hz,2H),1.25(t,J＝7.1Hz,3H),1.17(t,J＝7.1Hz,3H)。
Preparative example 19 synthesis of compound S19:
synthesis of Compound 19-2:
the synthesis of compound 19-2 was identical to that of compound 5-6, except that compound 19-1 was used instead of compound 5-4.
Synthesis of Compound 19-4:
compound 19-2 (645mg, 1.63mmol) was dissolved in anhydrous methanol (10 ml), and compound 19-3 (0.6 ml, 9.78mmol) was added and reacted with heating at 70 ℃ for 14 hours. The reaction solution is directly mixed with silica gel and loaded on a column, and the compound 19-4 is obtained after column purification.
Synthesis of Compounds 19-5:
compound 19-4 (500mg, 1.18mmol) was dissolved in Dichloromethane (DCM) (10 ml), and thionyl chloride (SOCl) was slowly added dropwise at 0 ℃ under nitrogen protection 2 ) (0.19ml, 2.67mmol), and then the reaction was allowed to warm to room temperature for 1 hour. After the reaction is completed, the reaction liquid is dried in a spinning mode to obtain a crude product compound 19-5.
Synthesis of Compounds 19-6:
the compound 19-5 was placed in ice water (10 ml), and a saturated sodium bicarbonate solution (NaHCO) was slowly added dropwise thereto with stirring at room temperature 3 ) pH was adjusted =8. Extracting with dichloromethane, collecting organic phase, mixing with silica gel, loading onto column, and purifying to obtain compound 19-6.
Synthesis of Compounds 19-7:
the synthesis of compound 19-7 was identical to that of compound 1-4, except that compound 19-6 was used instead of 1-3.
Synthesis of Compounds 19-8:
the synthesis of compounds 19-8 was identical to that of compounds 1-6, except that compounds 19-7 was used instead of 1-4.
Synthesis of Compounds 19-9:
the synthesis of compounds 19-9 was identical to that of compounds 1-7, except that compounds 19-8 were used instead of 1-6.
Synthesis of compound S19:
the synthesis of compound S19 was identical to that of compound S1, except that compounds 19-9 were substituted for compounds 1-7.
1 H NMR (400 MHz, methanol-d) 4 )δ7.47(s,1H),7.46(s,1H),7.07(s,2H),6.56(s,1H),6.50(s,1H),5.78–5.64(m,2H),4.89(d,J＝5.1Hz,2H),4.83(d,J＝5.3Hz,2H),4.58(q,J＝6.7Hz,2H),4.53(d,J＝7.1Hz,2H),4.47(t,J＝9.9Hz,2H),4.01(t,J＝9.5Hz,2H),3.79(t,J＝6.1Hz,2H),3.57(s,3H),3.55(t,J＝4.6Hz,4H),2.31(d,J＝7.5Hz,2H),2.27(d,J＝5.7Hz,4H),2.18(s,3H),2.16(s,3H),1.71–1.62(m,2H),1.36–1.32(m,3H),1.29(d,J＝7.1Hz,3H)。
Preparative example 20 synthesis of compound S20:
synthesis of Compound 20-2:
the synthesis of compound 20-2 was identical to that of compound 9-3, except that compound 20-1 was used instead of compound 4-1. [ Synthesis of Compound 20-1 reference CN109071514A ]
Synthesis of Compound 20-3:
the synthesis of compound 20-3 was identical to that of compound 1-7, except that compound 20-2 was used instead of compound 1-6.
Synthesis of compound S20:
the synthesis of compound S20 was identical to that of compound S1, except that compound 20-3 was substituted for compound 1-7.
1 H NMR(400MHz,Methanol-d 4 )δ8.36(s,1H),7.91(d,J＝1.6Hz,1H),7.66(d,J＝8.5Hz,1H),7.61(s,1H),7.31(s,1H),7.26(d,J＝8.5Hz,1H),6.56(s,1H),6.46(s,1H),5.96–5.84(m,1H),5.80–5.70(m,1H),4.94(d,J＝5.2Hz,2H),4.79(d,J＝5.6Hz,2H),4.58(q,J＝7.1Hz,2H),4.48(q,J＝7.1Hz,2H),3.92(t,J＝6.3Hz,2H),3.60(t,J＝4.8Hz,4H),2.37–2.32(m,2H),2.32–2.24(m,4H),2.18(s,3H),2.10(s,3H),1.75–1.66(m,2H),1.37–1.24(m,6H)。
Preparative example 21 synthesis of compound S21:
synthesis of Compound 21-1:
the synthesis of compound 21-1 was identical to that of compound 4-2, except that compound 3-1 was replaced with compound 8-1.
Synthesis of Compound 21-2:
the synthesis of compound 21-2 was identical to that of compound 1-7, except that compound 20-1 was used instead of compound 1-6.
Synthesis of compound S21:
the synthesis of compound S21 was identical to that of compound S1, except that compound 21-2 was used instead of compound 1-7.
1 H NMR(400MHz,Methanol-d 4 )δ7.51(d,J＝1.4Hz,1H),7.39(d,J＝1.3Hz,1H),7.16(d,J＝1.5Hz,1H),7.12(s,2H),7.09(d,J＝1.5Hz,1H),6.55(s,1H),6.45(s,1H),5.81–5.68(m,2H),4.88(s,2H),4.85(s,2H),4.56(q,J＝7.1Hz,2H),4.47(q,J＝7.1Hz,2H),3.79(t,J＝6.1Hz,2H),3.61(s,3H),3.57(t,J＝4.6Hz,4H),2.32–2.24(m,6H),2.17(s,3H),2.09(s,3H),1.69–1.59(m,2H),1.32(t,J＝7.1Hz,3H),1.26(t,J＝7.1Hz,3H)。
Experimental example 1: cell screening experiment for activating human interferon gene stimulating protein (STING) and promoting IFN-beta expression by using compound prepared by application
The detection method and the principle are as follows: THP1-Blue-ISG cells of human origin (Catalog # THP-ISG, invivoGen) express a Secreted Embryonic Alkaline Phosphatase (SEAP) reporter gene under the control of the ISG54 minimal promoter and an interferon-stimulated response element (IRF). When alkaline phosphatase is secreted outside the cell, the OD650 can be measured by a color reaction to reflect its content. When a compound is added to a cell, the compound activates an Interferon Stimulated Gene (ISG) pathway, such as a molecule in the type I interferon pathway, which promotes increased downstream basic phosphorylation secretion and increased absorbance of a chromogenic reaction.
The test method comprises the following steps:
1. addition compounds (including compound 1A): each well of a 96-well cell culture plate was added 20. Mu.L of a compound diluted with physiological saline at a concentration of 100. Mu.M, in 3 duplicate wells. The positive control compound was ADU-S100 (2 ',3' -c-di-AM (PS) 2 (Rp, rp), catalog # tlrl-nacda2r, invivogen) at a concentration of 100. Mu.M. To the non-medicated control group, 20. Mu.L of physiological saline containing 1% Dimethylsulfoxide (DMSO) was added.
2. Adding cells: THP1-Blue-ISG cell count, adjusting cell concentration to 5X 10 5 Cells were added at 180. Mu.l/well for incubation. Thus, the final volume of each test well was 200. Mu.L, DMSO content was 0.1%, and the test concentration of the compound was 10. Mu.M. The final concentration of the positive control compound ADU-S100 is 10 mu M, and the positive control compound ADU-S100 is incubated for 24 hours for detection; the blank group was added with 180. Mu.L of culture solution.
3. Detecting a color development reaction: after 24 hours, 20. Mu.l of the culture medium was transferred to a new 96-well plate, 200. Mu.l of color developing solution Quanti-Blue was added thereto, the plate was placed in an incubator at 37 ℃ and the OD650 value was measured after 0.5 to 2 hours.
4. Screening concentration of compound: 10 μ M.
5. And (4) analyzing results:
wherein the compound OD650 is the OD650 value of the compound of the present invention, the blank OD650 is the OD650 value of the medium, and the control OD650 is the OD650 value of the control group without the compound of the present invention (cell only and 0.1% DMSO).
6. And (4) evaluating the result: when the activation multiple (Fold change) is more than or equal to 2, the activation is effective.
The experimental results are as follows:
TABLE 1 Effect of partial Compounds on stimulation of human Interferon Gene in THP1 cells at 10. Mu.M concentration
From the above table 1, it can be seen that most of the compounds of the present application have the effects of activating the human interferon gene stimulating protein in THP1 cells and promoting IFN- β expression, wherein the activating effect of compounds S1, S4, S5, S9, S17, S19 and S21 on STING is greater than that of compound 1A, and some of the compounds have stronger effects than ADUS100. Therefore, the interferon protein can be used as a potential medicine for treating tumors, infectious diseases and the like related to the interferon gene stimulating protein.
Experimental example 2: water solubility test of Compound S1 hydrochloride and Compound 1A hydrochloride
Preparation of compound S1 hydrochloride and compound 1A hydrochloride: 100mg of Compound S1 or Compound 1A was dissolved in 2mL of a methanol solution, and 1mL of a 2M solution of hydrogen chloride in methanol was added thereto, followed by stirring at room temperature overnight. And (3) generating a large amount of white solid, adding methanol into the reaction liquid after spin-drying, then spin-drying, adding methanol, pulping, filtering and drying to obtain a white compound S1 or 1A hydrochloride.
Single-point method water solubility test:
precisely weighing the compound 1A, S hydrochloride, completely dissolving the compound in 1ml of purified water, testing the absorption peak area of the compound under 254nM by using HPLC, preparing a saturated aqueous solution of the compound 1A, S hydrochloride, testing the absorption peak area under the same condition, and multiplying the peak area ratio by the concentration of a standard point to obtain the water solubility of the compound hydrochloride.
In HPLC test, the concentration of the compound 1A hydrochloride standard solution was 2.26mg/mL, the amount of sample was 2. Mu.L, the concentration of the compound S1 hydrochloride standard solution was 4.81mg/mL, and the amount of sample was 2. Mu.L. The results of the water solubility test are shown in table 2 below.
Table 2. Water solubility test results for compound 1A and compound S1 hydrochloride:
the test results in table 2 show: the water solubility of compound S1 hydrochloride is about 22 times that of compound 1A hydrochloride, indicating that compound S1 hydrochloride of the present application has very excellent water solubility while having good STING agonistic activity.
Claims (9)
1. A class of heterocycle-substituted benzimidazole dimers represented by the following general formula I:
in the above general formula I, the compound of formula I,
R 1 -T 1 -、R 2 -T 2 -each independently is triazolyl, imidazolyl, 4,5-dihydrooxazolyl or-C (= O) NH 2 Provided that R is 1 -T 1 -and R 2 -T 2 -not simultaneously being-C (= O) NH 2 ；
R 3 Selected from H or C1-C3 alkoxy, R 4 Selected from C1-C3 alkoxy substituted by morpholinyl or hydroxy;
R 5 、R 6 each independently is C1-C3 alkyl which is unsubstituted or substituted by halogen.
2. The heterocycle-substituted benzimidazole dimer according to claim 1, or a pharmaceutically acceptable salt thereof,
wherein R is 1 -T 1 -、R 2 -T 2 -each independently is 1,2,4-triazolyl, imidazolyl, 4,5-dihydrooxazolyl, or-C (= O) NH 2 Provided that R is 1 -T 1 -and R 2 -T 2 Not simultaneously being-C(＝O)NH 2 ；
R 3 Selected from H or methoxy, R 4 Selected from propoxy substituted with morpholinyl or hydroxy;
R 5 、R 6 each independently being an ethyl group unsubstituted or substituted by halogen.
3. A heterocycle-substituted benzimidazole dimer or a pharmaceutically acceptable salt thereof, wherein the heterocycle-substituted benzimidazole dimer is selected from one of the following compounds:
4. the heterocycle-substituted benzimidazole dimer or a pharmaceutically acceptable salt thereof according to claim 1, wherein the heterocycle-substituted benzimidazole dimer is in its zwitterionic or isomeric form.
5. The heterocycle-substituted benzimidazole dimer or a pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt comprises a salt of the heterocycle-substituted benzimidazole dimer represented by formula I with: phosphoric acid, sulfuric acid, hydrochloric acid, acetic acid, tartaric acid, citric acid, malic acid, aspartic acid, or glutamic acid.
6. A pharmaceutical composition comprising the heterocycle-substituted benzimidazole dimer of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, and optionally, a pharmaceutically acceptable carrier.
7. Use of the heterocycle-substituted benzimidazole dimer or pharmaceutically acceptable salt thereof of any one of claims 1-5, or the pharmaceutical composition of claim 6, in the preparation of an interferon gene stimulating factor agonist.
8. The use according to claim 7, wherein the interferon gene stimulating factor agonist is for the treatment of a tumor or an infectious disease.
9. Use of a heterocycle-substituted benzimidazole dimer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, or a pharmaceutical composition according to claim 6, in the preparation of an immunological composition or vaccine adjuvant.
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