Application of trimethylbenzofuran piperidinone derivative in sensitization STING agonist anti-tumor treatment

Abstract

The invention discloses an application of a trimethylbenzofuran piperidinone derivative in sensitizing STING agonist antineoplastic treatment, and the derivative is (7-((3, 4-dihydroisoquinoline-2 (1H)-yl) sulfonyl)-3, 5, 6-trimethylbenzofuran-2-yl) (piperidin-1-yl) ketone. The invention also discloses an application of the trimethylbenzofuran piperidinone derivative in sensitizing STING agonist antineoplastic treatment, and the derivative is (7-((3, 4-dihydroisoquinoline-2 (1H)-yl) sulfonyl)-3, 5, 6-trimethylbenzofuran-2-yl) (piperidin-1-yl) ketone. In the prior art, through a strategy of knocking out STIM1 through CRISPR-Cas9, although cGAMP-mediated I-type interferon secretion is improved, intracellular calcium ion concentration is remarkably reduced due to irreversibility of knockout, calcium ion balance is destroyed, and processes of subsequent antigen cross presentation and the like are not facilitated. According to the present invention, the small molecule is developed to achieve the separation of STIM1 and STING, and the intracellular calcium ion steady state is maintained while the STING endoplasmic reticulum retention is released, such that the tumor treatment effect of the STING agonist is effectively enhanced, and the clinical immune response rate is expected to be significantly improved. Specifically, the invention provides a small molecule compound (7-((3, 4-dihydroisoquinoline-2 (1H)-yl) sulfonyl)-3, 5, 6-trimethylbenzofuran-2-yl) (piperidine-1-yl) ketone, which has excellent biological activity, can promote the separation of STIM1 and STING and promote the transport of STING from endoplasmic reticulum to a Golgi apparatus, and obviously improves the level of an STING agonist secreting IFN-beta.

Description

TECHNICAL FIELD

[0001] The present application relates to the application of a trimethylbenzofuran piperidinone derivative in sensitizing STING agonist anti-tumor therapy, in particular to a small molecule compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone and its application in promoting STING activation. BACKGROUND

[0002] Malignant tumors seriously threaten human health. In recent years, immunotherapy, including immune checkpoint blockers, cancer vaccines, and immune agonists, has become a new strategy for clinical tumor treatment. Among them, immune agonists activate the innate immune signaling pathway in antigen-presenting cells, restart and complete the cancer-immune cycle, thereby achieving the purpose of controlling and eliminating tumors.

[0003] As a popular target for the design and development of immune agonists, the stimulator of interferon genes (STING) in dendritic cells is a key bridge between innate and adaptive immunity, and achieves tumor killing by promoting the secretion of type I interferons. A number of tumor treatment programs based on STING agonists have also been developed for clinical trials, which have prolonged the survival of patients to some extent. However, the translocation of STING from the endoplasmic reticulum to the Golgi apparatus is a rate-limiting step for activation. During this process, the stromal interaction molecule 1 (STIM1) protein located on the endoplasmic reticulum binds to STING, severely limiting its translocation efficiency, resulting in a low response rate to STING agonist therapy in clinical patients. Studies have shown that the interaction between the two is mainly concentrated in the transmembrane domain fragment of their endoplasmic reticulum membranes. Therefore, how to relieve the endoplasmic reticulum retention mediated by STIM1 is the key to sensitizing STING activation. Clinical studies have also found that one patient carrying a STIM1 mutation that inhibits STIM1 expression showed elevated levels of multiple cytokines and interferon-stimulated genes, suggesting that STIM1 is a suitable target for intervention in sensitizing STING activation.

[0004] In fact, STIM1 is the sensor of endoplasmic reticulum calcium ion concentration, mainly combined with STIMG protein in the resting state, and after depletion of endoplasmic reticulum calcium ions, the conformation of STIM1 changes, and after oligomerization, it is displaced to the endoplasmic reticulum-membrane junction, combined with calcium release-activated calcium modulator 1 (ORAI1) on the cell membrane, and the two together constitute calcium release-activated calcium channels (CRAC), promoting the influx of calcium ions.

[0005] Calcium ions as messengers are widely involved in key links of the cancer-immune cycle, such as antigen cross-presentation and T cell killing of tumor cells, indicating that calcium ions are indispensable for STING agonists to exert efficacy. This means that if the separation of STIM1 and STING is induced by some means, but the function of STIM1 as a CRAC calcium channel is destroyed, leading to a significant change in intracellular calcium ion concentration, although it can promote the secretion of type I interferons, but it is not conducive to the subsequent process from STING activation to killing of tumor cells, making it difficult to achieve the ideal sensitization effect of STING activation. A typical example of this is the strategy of knocking out STIM1 by CRISPR-Cas9, although it enhances cGAMP-mediated type I interferon secretion, but the irreversibility of knockout leads to a significant decrease in intracellular calcium ion concentration, disrupting calcium ion balance, which is not conducive to subsequent processes such as antigen cross-presentation.

[0006] Therefore, it is necessary to develop small molecules to separate STIM1 and STING, to relieve the endoplasmic reticulum retention of STING while maintaining the homeostasis of intracellular calcium ions, thereby effectively enhancing the tumor treatment effect of STING agonists, and is expected to significantly improve the clinical immune response rate. SUMMARY

[0007] The prior art uses the strategy of knocking out STIM1 by CRISPR-Cas9, although it enhances cGAMP-mediated type I interferon secretion, but the irreversibility of knockout leads to a significant decrease in intracellular calcium ion concentration, disrupting calcium ion balance, which is not conducive to subsequent processes such as antigen cross-presentation. The present application develops small molecules to separate STIM1 and STING, to relieve the endoplasmic reticulum retention of STING while maintaining the homeostasis of intracellular calcium ions, thereby effectively enhancing the tumor treatment effect of STING agonists, and is expected to significantly improve the clinical immune response rate. Specifically, the purpose of the present application is to provide a small molecule compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone for use in sensitizing STING activation.

[0008] To achieve the above-mentioned purposes of the application, the technical solutions adopted by the application are as follows.

[0009] Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in STING activation, or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in preparation of a STING activation agent.

[0010] Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in sensitized STING activation, or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in preparation of a sensitized STING activation agent.

[0011] Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in promoting separation of STIM1 and STING, or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in preparation of a reagent for promoting separation of STIM1 and STING.

[0012] Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in reducing co-localization of STING and endoplasmic reticulum, relieving endoplasmic reticulum retention of STING, or promoting translocation of STING from endoplasmic reticulum to Golgi apparatus, or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in preparation of a reagent for reducing co-localization of STING and endoplasmic reticulum, relieving endoplasmic reticulum retention of STING, or promoting translocation of STING from endoplasmic reticulum to Golgi apparatus.

[0013] Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in preparation of a drug.

[0014] Use of (7-((3,4-dihydroisoquinoline-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in preparation of a tumor treatment drug.

[0015] Use of (7-((3,4-dihydroisoquinoline-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in preparation of a tumor treatment drug.

[0016] The application discloses a STING activating agent, a STING activating sensitizing agent, a medicine or a medicine sensitizing agent, and an active ingredient of the agents comprises (7-((3,4-dihydroisoquinoline-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone.

[0017] In the application, the chemical structural formula of (7-((3,4-dihydroisoquinoline-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone is shown as follows:

[0018] The application provides a preparation method of the compound, comprising the following steps: obtaining an intermediate of formula (III) by clasp and substituent modification on a raw material of formula (II), and then obtaining the compound (7-((3,4-dihydroisoquinoline-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone of formula (I) through oxidation and sulfonamide.

[0019] ; Formula (II).

[0020] ; Formula (III).

[0021] The application discloses that the small molecule compound (7-((3,4-dihydroisoquinoline-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone has excellent biological activity, can promote the separation of STIM1 and STING and promote the transportation of STING from the endoplasmic reticulum to the Golgi body, and significantly improves the level of STING agonist secretion IFN-beta.

[0022] Therefore, the application discloses application of the small molecule compound (7-((3,4-dihydroisoquinoline-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in treatment of tumors.

[0023] In particular, the small molecule compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone can improve the efficacy of existing STING activators, that is, it has a sensitization effect on existing STING activators.

[0024] The present application first discovers that the compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone has the function of relieving STING endoplasmic reticulum retention, has obvious sensitization effect on STING activation, and can be used as a STIM1 molecular glue. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 The nuclear magnetic resonance chart of the compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone.

[0026] Figure 2 The mass spectrum chart of the compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone.

[0027] Figure 3 a: DC2.4 cell STIM1 and STING immunofluorescence staining chart; b: STIM1 and STING co-localization rate statistics.

[0028] Figure 4 a: DC2.4 cell STING and endoplasmic reticulum immunofluorescence staining chart; b: STING and endoplasmic reticulum co-localization rate statistics; c: Effect of compound combined with SR-717 of different concentrations on IFN-β secretion; d: Effect of compound combined with SR-717 of different concentrations on IFN-β secretion.

[0029] Figure 5 The tumor growth curve of mice after intratumoral injection of compound combined with SR-717 in 4T1 orthotopic model. DETAILED DESCRIPTION

[0030] The present application first discovers that the compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone has the function of relieving STING endoplasmic reticulum retention, has obvious sensitization effect on STING activation, and can be used as a STIM1 molecular glue.

[0031] The following illustrates the technical progress of the present application through specific experiments, all raw materials used are existing products, specific preparation operations and performance tests, statistical analysis are conventional techniques, and animal experiments meet the relevant requirements of Suzhou University.

[0032] Synthesis of compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone The intermediate is obtained by subjecting the raw material to a clasp and a substituent modification, and then subjected to oxidation and sulfonamide to obtain compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone.

[0033]

[0034] DMF (100 mL) was placed in a 250 mL round-bottom flask under argon at 0 °C, and 1-(2-hydroxy-4,5-dimethylphenyl)ethan-1-one (1640 mg, 10 mmol, 1 equivalent) was added. 4-Bromoacetoacetate (1.336 mL, 12 mmol, 1.2 equivalents) was added to the mixture. The solution was refluxed at 100 °C overnight, then cooled at room temperature. To the solution, EtOAc (30 mL) and H2O (50 mL) were added. The mixture was poured into saturated aqueous NaHCO3solution (30 mL). The aqueous layer was extracted with DCM (20 mL x 3). The organic layer was collected, washed with saturated aqueous Na2CO3solution, dried over Na2SO4, and the solvent was removed in vacuo. The crude product was purified by column chromatography (AcOEt / cyclohexane, 1 / 100 to 1 / 50) to give 3,5,6-trimethylbenzofuran-2-carboxylic acid ethyl ester (1612 mg, 6.94 mmol, 69%) as a white solid.

[0035] To a mixture of ethyl 3,5,6-trimethylbenzofuran-2-carboxylate (1612 mg, 1.0 eq), piperidine (707.88 mg, 1.2 eq), toluene (300 mL, 15.0 vol) was added LiHMDS (2317 mg, 2 eq) at room temperature. The mixture was stirred at room temperature for 15 hours and the progress of the reaction was monitored by TLC. Upon completion, the mixture was poured into cooled H2O (600 mL, 30 vol). The aqueous layer was extracted with AcOEt (20 mL x 3). The organic layer was collected, washed with saturated aqueous NaCl, dried over Na2SO4, and the solvent was removed in vacuo. The crude product was purified by column chromatography (AcOEt / cyclohexane, 1 / 20) to give piperidin-1-yl(3,5,6-trimethylbenzofuran-2-yl)methanone (813.45 mg) as a white solid.

[0036] Compound piperidin-1-yl(3,5,6-trimethylbenzofuran-2-yl)methanone (813.45 mg, 1.0 eq), NBS (N-bromosuccinimide, 533.94 mg, 1.0 eq) were added in 50 mL CH3CN in a 100 mL single necked flask. The mixture was stirred at 80 °C overnight and the progress of the reaction was monitored by TLC. To the mixture was added saturated aqueous NaCl (30 mL). The aqueous layer was extracted with DCM (10 mL x 3). The organic layer was collected, washed with saturated aqueous NaCl, dried over Na2SO4, and the solvent was removed in vacuo. The crude product was purified by column chromatography (AcOEt / cyclohexane, 1 / 100 to 1 / 50) to give (7-bromo-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone (348 mg) as a white solid.

[0037] (7-bromo-3,5,6-trimethylbenzofuran-2-yl)(piperidin-l-yl)methanone (348 mg, 1 equiv), benzyl mercaptan (186 mg, 1.5 equiv), DIPEA (N,N-diisopropylethylamine, 580 mg, 3 equiv), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (173 mg, 0.2 equiv), Pd2(dba)3 (137 mg, 0.1 equiv) were added in 50 mL of Dioxane (1,4-dioxane) in a 100 mL single necked flask. The mixture was stirred at 100 °C for 2 h and the progress of the reaction was monitored by TLC. To the mixture was added saturated aqueous NaCl (30 mL). The aqueous layer was extracted with DCM (20 mL x 3). The organic layer was collected, washed with saturated aqueous NaC104, dried over Na2S04and the solvent was removed in vacuo. The crude product was purified by column chromatography (AcOEt / cyclohexane, 1 / 20 to 1 / 10) to give (7-(benzylthio)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-l-yl)methanone (390 mg) as a white solid.

[0038] To a mixture of compound (7-(benzylthio)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-l-yl) (390 mg, 1 equiv) in a mixture of AcOH / H20 (1:1) was added NCS (N-chlorosuccinimide, 400 mg, 3 equiv) and the reaction was allowed to proceed at room temperature for 2 h. The reaction was quenched with H20 and the aqueous layer was extracted with DCM (10 mL x 3). The organic layer was collected, dried over Na2S04and the solvent was removed in vacuo. This gave 3,5,6-trimethyl-2-(piperidin-l-ylcarbonyl)benzofuran-7-sulfonyl chloride as a light yellow solid. To the crude compound 3,5,6-trimethyl-2-(piperidin-l-ylcarbonyl)benzofuran-7-sulfonyl chloride was added 30 mL of DMF, pyridine (5 equiv), tetrahydroisoquinoline (1 equiv). The mixture was heated to 35 °C on a silicon oil bath and stirred for 3 h. TLC was checked until the reaction was complete. H20 was added to quench. The product was extracted and washed with ethyl acetate and water solution, concentrated and purified by column separation to give (7-((3,4-dihydroisoquinolin-2(lH)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-l-yl)methanone (215 mg); its NMR data are as follows: 1 H NMR (600 MHz, DMSO- d 6) δ 7.57 (s, 1H), 7.22 - 7.15 (m, 3H), 7.11 (ddd, J= 7.5, 5.8, 3.5Hz, 1H), 4.42 (s, 2H), 3.63 (t, J = 5.7 Hz, 2H), 3.55 (dt, J = 15.8, 5.4 Hz,4H), 2.86 (td, J = 5.8, 0.9 Hz, 2H), 2.52 (s, 3H), 2.48 (s, 3H), 2.35 (s,3H), 1.71 (p, J = 5.6 Hz, 4H), 1.56 (p, J = 5.7 Hz, 2H)。 Figure 1 NMR chart of compound (7-((3,4-dihydroisoquinolin-2(lH)-yl)sulfonyl)-3,5,6- trimethylbenzofuran-2-yl)(piperidin-l-yl)methanone. Figure 2 Mass spectrum chart of compound (7-((3,4-dihydroisoquinolin-2(lH)-yl)sulfonyl)-3,5,6- trimethylbenzofuran-2-yl)(piperidin-l-yl)methanone.

[0039] Example 1 Study on compound (7-((3,4-dihydroisoquinolin-2(lH)-yl)sulfonyl)-3,5,6- trimethylbenzofuran-2-yl)(piperidin-l-yl)methanone promoting the separation of STIM1 and STING The effect of compound on the co-localization of STIM1 and STING was investigated by immunofluorescence staining. After compound (5 μM) combined with STING agonist SR-717 (30 μg mL -1 ) was administered to incubate mouse DC2.4 cells for 2 h, the drug was discarded, and the cells were fixed with 4% paraformaldehyde, followed by permeation with 0.1% Triton X-100 for 15 min and blocking with 1% BSA for 1 h. STIM1 primary antibody was added for incubation at 4°C overnight, the STIM1 primary antibody was discarded, the cells were washed with PBS for 3 times, Alexa Fluor 594 conjugated secondary antibody was added for staining at 37°C for 90 min in the dark, the secondary antibody was discarded, the cells were washed with PBS for 3 times, Alexa Fluor 488 conjugated STING antibody was added for incubation at 4°C overnight, the STING antibody was discarded, the cells were washed with PBS for 3 times, and the cell nuclei were stained with Hoechst33342 solution. The co-localization of STIM1 and STING was photographed by laser confocal microscope, and the co-localization rate of STIM1 and STING was calculated by Image J software. The experimental results showed that (Fig. 6) Figure 3 ): the compound reduced the co-localization of STIM1 and STING Figure 3a), the co-localization rate further decreased when combined with SR-717 Figure 3 b), suggesting that the compound promoted the separation of STIM1 and STING.

[0040] Example Two Sensitization activity of compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone on STING activation The effect of the compound on the co-localization of STING and endoplasmic reticulum after administration was investigated by immunofluorescence staining. After the mouse DC2.4 cells were incubated with the compound (5 μM) combined with the STING agonist SR-717 for 2 h, the drug was discarded, and the cells were fixed with 4% paraformaldehyde, followed by permeation with 0.1% Triton X-100 for 15 min and blocking with 1% BSA for 1 h. CRT (endoplasmic reticulum marker) primary antibody was added and incubated at 4°C overnight, the CRT primary antibody was discarded, and the cells were washed with PBS for 3 times, Alexa Fluor 594-conjugated secondary antibody was added, and the cells were stained at 37°C in the dark for 90 min, the secondary antibody was discarded, and the cells were washed with PBS for 3 times, Alexa Fluor 488-conjugated STING antibody was added and incubated at 4°C overnight, the STING antibody was discarded, and the cells were washed with PBS for 3 times, and the cell nuclei were stained with Hoechst33342 solution. The co-localization of STING and endoplasmic reticulum was photographed by laser confocal microscope, and the co-localization rate of STING and endoplasmic reticulum was calculated by Image J software. The experimental results showed that Figure 4 a-b): The compound significantly reduced the co-localization of STING and endoplasmic reticulum, indicating that it relieved the endoplasmic reticulum retention mediated by STIM1 and promoted the translocation of STING from endoplasmic reticulum to Golgi.

[0041] The interferon-β (IFN-β) enzyme-linked immunosorbent assay kit adopts a competitive ELISA method. The mouse bone marrow cells extracted in the early stage are induced into BMDC cells with 20 ng / mL granulocyte-macrophage colony stimulating factor (GM-CSF), inoculated into a 48-well culture plate before the experiment, 0.3125-20 µM compounds and 30 μg / mL STING agonist SR-717, or 5 µM compounds and 20-40 μg / mL SR-717 are added to each well, and the supernatant is collected after 12 h of reaction for detection of interferon-β (IFN-β) content. The IFN-β antigen is coated on the enzyme-labeled plate, and the IFN-β in the sample or standard competes with the coated IFN-β for the binding sites on the biotin-labeled anti-IFN-β monoclonal antibody during the experiment, and the free components are washed away. Add horseradish peroxidase-labeled avidin, and biotin and avidin specifically bind to form an immune complex, and the free components are washed away. Add color developing substrate (TMB), and TMB turns blue under the catalysis of horseradish peroxidase, and turns yellow after the addition of a stop solution. The OD value is measured at 450 nm wavelength by an enzyme-labeled instrument, and the concentration of IFN-β in the sample is calculated by drawing a standard curve. The experimental results show that the compound itself cannot promote the secretion of IFN-β, and when different concentrations of the compound are combined with SR-717, the sensitization effect of the compound shows a concentration-dependent manner, and gradually reaches a platform as the concentration increases Figure 4 c). When the compound is combined with different concentrations of SR-717 at a fixed concentration, the compound can improve the ability of different concentrations of SR-717 to secrete IFN-β and the upper limit Figure 4 d).

[0042] Example Three Influence of Compound (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone combined with STING agonist on anti-tumor effect Logarithmic growth period 4T1 cells were taken, counted after digestion, resuspended with RPMI1640 incomplete medium, inoculated in the mammary fat pad of Balb / c mice (1.5×10 6 cells per mouse, 50 μL per mouse), and a 4T1 orthotopic tumor model was constructed. When the tumor volume grew to about 100 mm 3 , it was divided into PBS, compound, SR-717, compound+SR-717 groups (n=5). The dose of SR-717 was 30 mg kg -1 , the dose of the compound was 1 mg kg -1The drug was injected into the mice intratumorally every 2 days for 5 times. The beginning of the drug administration was recorded as day 0, and the size of the tumor of the mice was measured within 30 days after the drug administration to calculate the tumor volume (V = long diameter x short diameter 2 / 2). When the tumor volume of the mice reached 1500 mm 3 , the mice were sacrificed, and the tumor growth curve was monitored. The experimental results showed that the compound itself had no obvious tumor inhibition effect, SR-717 could delay the tumor growth trend to a certain extent, and the tumor inhibition effect was further improved when combined with the compound, indicating that the compound significantly improved the anti-tumor effect of the STING agonist. Figure 5 ​

Claims

1. Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the activation of STING, or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the preparation of a STING activation agent.

2. Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the sensitization of STING activation, or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the preparation of a sensitized STING activation agent.

3. Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the promotion of the dissociation of STIM1 from STING, or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the preparation of a STIM1 dissociation from STING promoting agent.

4. Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the reduction of the co-localization of STING with the endoplasmic reticulum, the release of STING from endoplasmic reticulum retention, or the promotion of the translocation of STING from the endoplasmic reticulum to the Golgi apparatus; or use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the preparation of a STING co-localization with the endoplasmic reticulum reduction, STING release from endoplasmic reticulum retention, or STING translocation from the endoplasmic reticulum to the Golgi apparatus promoting agent.

5. Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the preparation of a medicament.

6. Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the preparation of a tumor treatment drug.

7. Use of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone in the preparation of an agent for improving the therapeutic effect of a tumor drug.

8. Use according to any one of claims 1 to 7, characterized in that, The chemical structure of (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone is as follows: 。 9. A STING-activating agent, a STING-activating sensitizing agent, a drug, or a drug-sensitizing agent, characterized in that, The active ingredient comprises (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone.

10. A process for the preparation of (7-((3,4-dihydroisoquinolin-2(lH)-yl)sulfonyl)-3,5,6- trimethylbenzofuran-2-yl)(piperidin-l-yl)methanone, characterized in that, The process comprises the following steps: oxidation, sulphonamidation of an intermediate of formula (III) to obtain (7-((3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-3,5,6-trimethylbenzofuran-2-yl)(piperidin-1-yl)methanone; the chemical structure of the intermediate of formula (III) is shown below: 。

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