CN112442033A

CN112442033A - STING pathway modulators and uses thereof

Info

Publication number: CN112442033A
Application number: CN201910803609.5A
Authority: CN
Inventors: 赵海峰
Original assignee: Medshin International Co ltd
Current assignee: Medshin International Co ltd
Priority date: 2019-08-28
Filing date: 2019-08-28
Publication date: 2021-03-05
Also published as: WO2021037179A1

Abstract

The present invention relates to compounds of formula I or pharmaceutically acceptable salts, solvates, or prodrugs thereof. The invention also relates to a preparation method of the compound or the pharmaceutically acceptable salt, solvate or prodrug thereof, a medicinal composition containing the compound, a method for treating and preventing diseases related to the STING pathway and a pharmaceutical application of the compound.

Description

STING pathway modulators and uses thereof

Technical Field

The present invention relates to compounds that modulate STING activity and their use in treating diseases. The invention also relates to a preparation method of the compound or the pharmaceutically acceptable salt, the solvate or the prodrug thereof, a medicinal composition containing the compound and application of the compound or the pharmaceutically acceptable salt, the solvate or the prodrug thereof in preparing medicines.

Background

The innate immune response is the first line of defense of the host against foreign pathogenic microorganisms. The host can recognize pathogen-associated molecular patterns via pattern recognition receptors, which in turn initiate an antiviral immune response. Interferon activating protein (STING) is an important molecule in the innate immune response, which plays an important role in the defense against viral and intracellular bacterial infections, mediating the production of type I interferons. STING can either directly recognize cyclic-di-nucleotides or act as an adaptor protein to facilitate transduction of downstream signaling pathways; STING serves as an important component of the signal transduction cascade and plays an immune defense role in the infection by pathogens (viruses, bacteria, parasites, etc.). After STING is activated, the production of interferon and cytokines can be increased, and an adaptive immune system is activated through a series of cascade reactions, so that T cells are activated.

Whether the DNA is from a virus, a bacterium, or the organism itself, it binds to an enzyme called cyclic GMP-AMP synthsase (cGAS), which links two nucleotides to form a cyclic dinucleotide called cGAMP, thereby activating STING. DNA in humans does not normally activate the STING protein, since DNA can normally be present in the nucleus (except for mitochondrial DNA). However, if the DNA leaks into the cytoplasm, it activates STING, triggering an immune response.

In addition to combating infection by foreign pathogens, STING agonists also play an important role in the immunotherapy of tumours. Tim Mitchison's laboratory at Harvard medical college studied a STING agonist-DMXAA, which was extremely potent in a mouse anti-tumor model. However, DMXAA can only bind to STING protein of mice, which restricts the subsequent clinical application.

Cyclic Dinucleotides (CDNs) are natural agonists of STING and researchers at adoro Biotech found that direct injection of synthetic CDNs into mice tumours could activate STING, thereby activating the innate immune system and triggering a cascade of responses and activating T cells against tumours. Aduro's research team also injected more tumor cells into mice, spreading them throughout the lungs. Unexpectedly, a single intratumoral injection of CDN could activate T cells systemically in mice and clear tumors elsewhere, including the lungs. If the results were reproducible in patients, this would mean that STING agonists would have very bright prospects, since most patients ultimately die not from carcinoma in situ, but from tumors formed by metastasis of cancer cells.

At present, STING agonists are in the infancy stage, and studies of agonists have focused mainly on work on modifying natural Cyclic Dinucleotide (CDNs) analogues. However, the CDNs compounds are limited by low membrane permeability and metabolic instability and are difficult to effectively utilize clinically. Research on non-CDNs small molecules is carried out at the same time, mainly comprising 5 ', 6' -dimethylxanthene-4-acetic acid (DMXAA) and 10-carboxyymethyl-9-acridanone (CMA), and the difference of species selectivity is a main factor influencing further development of the molecules. At present, the human agonists STING that have been identified mainly include: g10, DSDP, C11, BNBC and alpha-mangostin, all of which have antiviral or antitumor activity but induce less immune responses than cyclic dinucleotides. In 2018 researchers developed dimers of aminobenzenzimidazole (abzi): trihydrochloride is a strong non-CDNs STING agonist which is publicly reported, but the molecular weight is too large, and the drugability is uncertain.

The existing research results show that STING is a significant drug target, and the anti-tumor, anti-virus and anti-bacterial effects can be realized by activating the STING pathway. The aim of the present invention is to develop novel agonists of the STING pathway for the preparation of novel therapeutic agents for broad-spectrum antiviral, antibacterial or cancer immunotherapy.

Disclosure of Invention

The inventors have unexpectedly found that the compounds of formula I as defined below are capable of effectively modulating the activity of the STING pathway, thereby providing the use and pharmaceutical compositions of said compounds for the treatment of STING-related diseases.

In particular, the present invention provides compounds of formula I as follows:

wherein the content of the first and second substances,

R₁、R₂and R₃Independently selected from hydrogen, alkyl, alkoxy, hydroxy,

r4 and R5 are independently selected from hydrogen, alkoxy, acetyl, or R4 or R5 are linked to the N atom on the adjacent pyridine ring to form ═ N-C (═ O) -C-.

In one aspect of the invention, the compound of formula I is selected from:

9, 10-dimethyl-6H-pyrrolo [3,2,1-de ] -1, 6-acridinedione,

2- (5, 6-dimethyl-9-acridone) -4-acetic acid,

2- (5, 6-dimethyl-7-methoxy-9-acridone) -4-acetic acid,

2-methoxy-5, 6-dimethyl-9-acridone-4-carboxylic acid,

3, 4-dimethyl-9-acridone,

2-methoxy-5, 6-dimethyl-9-acridone,

2-methoxy-3, 4-dimethyl-9-acridone, and

2, 7-dimethoxy-3, 4-dimethyl-9-acridone.

Wherein the alkyl group may be a lower alkyl group, for example a 1-8C alkyl group. It includes straight-chain and branched alkyl groups such as methyl, ethyl, propyl, isopropyl, tert-butyl and the like, and also (3-8C) cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.

Wherein said alkoxy may be a lower alkyloxy, e.g. 1-8C alkoxy. Including methoxy, ethoxy, propoxy, isopropoxy, butoxy, and the like.

In yet another aspect of the invention, the compound of formula I is 2- (5, 6-dimethyl-7-methoxy-9-acridone) -4-acetic acid.

In yet another aspect of the present invention, the compound of formula I is 2-methoxy-5, 6-dimethyl-9-acridone.

The invention also provides a preparation method of the compound shown in the formula I.

In one aspect of the invention, the process for preparing the compound of formula I comprises:

for example, the compounds of formula i and formula ii are used as starting materials, and are heated to 110 ℃ in N, N-dimethylformamide containing a mixed catalyst of copper powder and cuprous oxide,coupling to produce a compound of formula iii, heating and stirring the compound of formula iii in a proper amount of Eton's reagent, and condensing to produce a compound of formula I, wherein R₁、R₂、R₃、R₄、R₅Is as defined above.

Suitable pharmaceutically acceptable salts of the compounds of formula I according to the invention are, for example, acid addition salts of the compounds of formula I, for example with inorganic or organic acids, such as hydrochloric acid, hydrobromic acid, sulphuric acid, trifluoroacetic acid, citric acid or maleic acid; or an ammonium salt, or a salt with an organic base such as methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris- (2-hydroxyethyl) amine, or an alkali metal.

Suitable pharmaceutically acceptable solvates of the compounds of formula I according to the invention are, for example, hydrates, such as hemihydrate, monohydrate, dihydrate or trihydrate, or alternatively combined amounts thereof.

The compounds of the invention may be administered in the form of prodrugs, which are modified forms of prodrugs well known in the art that cleave in the human or animal body to release the compounds of the invention. Examples of prodrugs include in vivo cleavable ester derivatives, which may be formed at the carboxy or hydroxy group of the compounds of formula I, and in vivo cleavable amide derivatives, which may be formed at the carboxy or amino group of the compounds of formula I.

Pharmaceutically acceptable prodrugs of compounds of formula I may also be in the form of in vivo cleavable esters or ethers, examples including in vivo cleavable esters or ethers of compounds of formula I containing a hydroxyl group for which suitable pharmaceutically acceptable ester-forming groups include inorganic esters such as phosphate esters (including cyclic phosphoramidate esters). For hydroxy, suitable pharmaceutically acceptable ester-forming groups also include (1-10C) alkanoyl groups (e.g., acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl), (1-10C) alkoxycarbonyl groups (e.g., ethoxycarbonyl, N- [ di- (1-4C) alkyl ] carbamoyl, 2-dialkylaminoacetyl and 2-carboxyacetyl). Examples of the ring substituents on the phenylacetyl group and the benzoyl group include aminomethyl group, N-alkylaminomethyl group, N-dialkylaminomethyl group, morpholinomethyl group, piperazin-1-ylmethyl group and 4- (1-4C) alkylpiperazin-1-ylmethyl group. For hydroxy, suitable pharmaceutically acceptable ether-forming groups include-acyloxyalkyl groups such as acetoxymethyl and pivaloyloxymethyl.

Pharmaceutically acceptable prodrugs of compounds of formula I may also be, for example, in vivo cleavable amide derivatives thereof. Suitable pharmaceutically acceptable amides formed from amino groups include, for example, amides formed with (1-10C) alkanoyl groups such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl. Examples of the ring substituents on the phenylacetyl group and the benzoyl group include aminomethyl group, N-alkylaminomethyl group, N-dialkylaminomethyl group, morpholinomethyl group, piperazin-1-ylmethyl group and 4- (1-4C) alkylpiperazin-1-ylmethyl group.

The compounds of formula I of the present invention, or pharmaceutically acceptable salts, solvates or prodrugs thereof, have STING agonistic activity. Therefore, the compound of formula I or the pharmaceutically acceptable salt, solvate or prodrug thereof can be used for preventing and/or treating the diseases related to the STING pathway or preparing medicaments for preventing and/or treating the diseases related to the STING pathway, such as the diseases treated by STING activation or preparing the medicaments for treating the diseases by STING activation.

In one aspect of the present invention there is provided a pharmaceutical composition comprising a compound of formula I as hereinbefore defined, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in admixture with a pharmaceutically acceptable excipient, for example a diluent or carrier.

The pharmaceutical compositions of the present invention can be obtained by conventional methods using conventional pharmaceutical excipients well known in the art. For example, compositions intended for oral use may contain excipients which are one or more coloring, sweetening, flavoring and/or preservative agents and the like. As used herein, the term "pharmaceutically acceptable carrier, diluent or excipient" refers to a pharmaceutically acceptable excipient commonly used in the formulation industry, such as those listed in Rotomong et al, pharmaceutical excipients university, Sichuan scientific technology Press, 1995.

The pharmaceutical compositions of the present invention can be obtained by conventional methods using conventional pharmaceutical excipients well known in the art. For example, compositions intended for oral use may contain excipients which are one or more coloring, sweetening, flavoring and/or preservative agents and the like.

The pharmaceutical compositions of the invention may take a form suitable for oral administration (e.g., tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (e.g., creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (e.g., finely divided powders or liquid aerosols), for administration by insufflation (e.g., finely divided powders), or for parenteral administration (e.g., sterile aqueous or oily solutions for intravenous, subcutaneous, intraperitoneal or intramuscular administration or suppositories for rectal administration).

The amount of active ingredient combined with one or more excipients to form a single dosage form will vary as desired, depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans will typically contain, for example, from 0.01mg to 1g (more suitably from 0.1 to 250mg, for example from 0.1 to 100mg) of the active ingredient in admixture with a suitable and convenient amount of excipient which may be present in the range of from about 5 to 98% by weight of the total composition.

The size of the dose of a compound of formula I, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for therapeutic or prophylactic purposes will naturally vary according to the nature and severity of the disease, the age and sex of the animal or patient and the route of administration, according to well-known principles of medicine.

According to a further aspect of the present invention there is provided the use of a compound of formula I as described hereinbefore, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the manufacture of a medicament for the treatment or prophylaxis of a disease associated with the STING pathway.

According to yet another aspect of the present invention, the STING pathway-related disease is an infectious disease, cancer, inflammation, allergy, and autoimmune disease.

The infectious disease is a viral infectious disease or a bacterial infectious disease, and the infectious agents include viral pathogens such as influenza virus, enterovirus, hepatitis B virus, hepatitis C virus, Ebola virus, Marburg virus, SARS virus, Securia virus, bunyavirus, rhinovirus, respiratory nucleopolyvirus, cholera virus, and the like, and bacterial pathogens such as tubercle bacillus, Escherichia coli, Acinetobacter baumannii, Diplococcus pneumoniae, Streptococcus lactis, Sarcina ureabanica, Staphylococcus aureus, Bacillus subtilis, Bacillus anthracis, Bacillus subtilis, Streptococcus, Proteus, Vibrio cholerae, Treponema pallidus, and the like.

The cancer is gastric cancer, liver cancer, lung cancer, esophageal cancer, cervical cancer, breast cancer, colon cancer, rectal cancer, nasopharyngeal cancer, ovarian cancer, renal cancer, bladder cancer, thyroid cancer, skin cancer, etc.; malignant tumors derived from mesenchymal tissues such as muscle, fat, bone, blood vessel, lymph, etc., for example, rhabdomyosarcoma, leiomyosarcoma, fibrosarcoma, liposarcoma, osteosarcoma, chondrosarcoma, angiosarcoma, lymphosarcoma, etc. Also, for example, leukemia, Hodgkin's disease, Wilms ' tumor (nephroblastoma), melanoma, retinoblastoma, seminoma, granuloma, Kupffer's tumor, Ewing's tumor, malignant vascular endothelial cell tumor, or Paget's disease of the breast. Preferably digestive tract cancer and respiratory tract cancer, such as gastric cancer, hepatocarcinoma, lung cancer, esophageal cancer, and carcinoma of large intestine.

Inflammation represents the vascular, cellular, and neural response to trauma. Inflammation can be characterized by the movement of inflammatory cells, such as monocytes, neutrophils, and granulocytes, into the tissue. This often involves reduced endothelial barrier function and edema entry into the tissue. Inflammation can be classified as acute or chronic. Acute inflammation is the initial response of the body to noxious stimuli and is achieved by an increase in the movement of plasma and leukocytes from the blood to the damaged tissue. The cascade of biochemical events spreads and matures the inflammatory response, which involves damaging the local vasculature, immune system and various cells within the tissue. Chronic inflammation, known as chronic inflammation, results in progressive changes in the cell types present at the site of inflammation and is characterized by the simultaneous destruction and healing of tissues from the inflammatory process. Inflammation includes musculoskeletal inflammation, vascular inflammation, neuroinflammation, inflammation of the digestive system, inflammation of the eye, inflammation of the reproductive system, or other inflammation, such as asthma.

Autoimmune diseases include, but are not limited to, systemic lupus erythematosus, psoriasis, Insulin Dependent Diabetes Mellitus (IDDM), dermatomyositis, and Sjogren's Syndrome (SS).

Allergic diseases include, but are not limited to, allergic reactions, contact dermatitis (including that caused by poison ivy), urticaria, skin allergies, respiratory allergies (hay fever, allergic rhinitis), and gluten-sensitive bowel disease (celiac disease).

Detailed Description

The present invention will be further described by the following examples and test examples. However, the scope of the present invention is not limited to the following examples or experimental examples. It will be understood by those skilled in the art that various changes and modifications may be made to the invention without departing from the spirit and scope of the invention. The present invention has been described generally and/or specifically with respect to materials used in testing and testing methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is nevertheless described herein in as detail as possible.

For all of the following examples, standard procedures and purification methods known to those skilled in the art may be used. Unless otherwise indicated, all temperatures are expressed in degrees Celsius. The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or Mass Spectrometry (MS). The melting point m.p. of the compound was determined by YRT-3 drug melting point apparatus, the thermometer was uncorrected, and m.p. is given in ° C.¹H NMR was measured by a model AVANCE III HD-400 NMR spectrometer. Mass spectra were determined by an AB Sciex API model 4000 mass spectrometer. All solvents for the reaction are not indicated to have been subjected to a standardized pretreatment.

In the following examples,% means mass% unless otherwise specified.

Example preparation of 110-dimethyl-6H-pyrrolo [3,2,1-de ] -1, 6-acridinedione

Reacting 2-bromobenzoic acid(2.16g,10.00mmol), 2-amino-3, 4-dimethylbenzoic acid (1.5g,9.1mmol), copper powder (0.06g) and cuprous oxide (0.06g) are added into N, N-dimethylformamide (18ml), materials are uniformly mixed at about 50 ℃, the temperature is increased to 100 ℃ and 110 ℃ for reaction overnight, the reaction progress is monitored by thin layer chromatography, after the reaction is completed, the reaction liquid is kept stand and cooled to room temperature, and then the N, N-dimethylformamide is evaporated under reduced pressure to obtain a dark brown solid. 1N sodium hydroxide solution (60ml) was added to dissolve, and copper powder was removed by filtration to give a yellow-brown clear filtrate. Concentrated hydrochloric acid was slowly added dropwise to the filtrate to a solution pH of 5, filtered after thorough stirring and washed with water to give a pale yellow solid, which was dried and weighed as 1.81g, yield 66.7%. Adding the light yellow solid into 9ml of Eton reagent, heating and stirring at 90-100 ℃ for 3.5h, standing and cooling to room temperature, slowly pouring into 150ml of saturated sodium bicarbonate solution, and fully stirring after the addition is finished to precipitate a large amount of solid. Filtration and washing with copious amounts of water gave crude product which was dried and purified by silica gel column chromatography to give a white solid weighing 1.52g dry with a total yield of 63.3%.¹H NMR(400MHz,DMSO-D6)δ：8.11(d,J＝8.0Hz,1H),7.97(d,J＝8.1Hz,1H),7.75(d,J＝7.0Hz,1H),7.51–7.40(m,2H),4.05(s,2H),3.37(s,1H),2.43(s,3H),2.33(s,3H).ESI-MS(m/z)：264.1[M+H]+。

Example 22 preparation of (5, 6-dimethyl-9-acridone) -4-acetic acid

9, 10-dimethyl-6H-pyrrolo [3,2,1-de ] is obtained as described in example 1 above]And (3) after the 1, 6-acridinedione is added, 0.8g of the product is put into 40ml of ethanol solution, after the uniform mixing, 8ml of NaOH solution with the concentration of 1N is added, the temperature is increased to 100 ℃, the reaction is carried out for 4.5h, the reaction solution is dissolved by 40ml of water after the decompression and the spin drying, and then 2N hydrochloric acid is added to acidify to the PH value of 7, so that a large amount of solid is separated out. Fully stirring and filtering to obtain a crude product. After vacuum drying, purification by silica gel column chromatography gave a yellow solid weighing 0.70g on dry basis with a yield of 83.2%.¹H NMR(400MHz,DMSO-D6)δ:13.11(s,1H),9.53(s,1H),8.14(dd,J＝8.1,1.3Hz,1H),8.05(d,J＝8.2Hz,1H),7.66(dd,J＝7.2,1.5Hz,1H),7.27(m,J＝8.0,7.2Hz,1H),7.13(d,J＝8.3Hz,1H),4.10(s,2H),2.52(s,3H),2.41(s,3H).13C NMR(101MHz,DMSO-D6)δ:177.44(s),173.65(s),142.28(s),139.63(s),139.51(s),135.88(s),125.76(s),124.72(s),123.69(s),123.23(s),121.46(s),121.24(s),119.31(s),38.59(s),21.15(s),12.63(s).MS(ESI):m/z 282.11[M+H]+。

Example preparation of 32- (5, 6-dimethyl-7-methoxy-9-acridone) -4-acetic acid

After the starting material, 2-amino-3, 4-dimethylbenzoic acid, was replaced with 2-amino-3, 4-dimethyl-5-methoxybenzoic acid by the method in example 1 above and the corresponding intermediates were charged in proportion, 0.70g of a gray solid was obtained in 27.7% overall yield by the method in example 6 above.¹H NMR(400MHz,DMSO-D6)δ:9.46(s,1H),8.18(dd,J＝8.1,1.3Hz,1H),7.68(dd,J＝7.2,1.3Hz,1H),7.52(s,1H),7.24(dd,J＝8.0,7.2Hz,1H),4.13(s,2H),3.86(s,3H),2.51(s,3H),2.36(s,3H).13C NMR(101MHz,DMSO-D6)δ:176.68,173.72,153.32,139.26,135.57,134.29,133.22,125.76,125.31,123.21,121.45,120.71,119.29,101.53,55.88,38.69,13.40,13.19.MS(ESI):m/z 312.14[M+H]+。

EXAMPLE 42 preparation of methoxy-5, 6-dimethyl-9-acridone-4-carboxylic acid

The starting material, 2-bromobenzoic acid, was replaced by 2-bromo-5-methoxybenzoic acid by the method described in example 1 above, and the batch was charged in proportion to give 0.32g of green solid with a total yield of 45.2%.¹H NMR(400MHz,DMSO-D6)δ:14.09(s,1H),11.83(s,1H),7.99–7.83(m,3H),7.14(d,J＝8.2Hz,1H),3.82(s,3H),2.41(s,3H),2.33(s,3H).13C NMR(101MHz,DMSO-D6)δ:173.72,168.61,148.83,138.47,136.65,133.48,121.22,120.77,120.48,120.09,118.17,117.44,116.13,111.52,59.12,18.17,10.66.MS(ESI):m/z 298.11[M+H]+。

Example 53 preparation of 4, 4-dimethyl-9-acridone

The starting materials, 2-bromophenylacetic acid and 2-amino-3, 4-dimethylbenzoic acid, were replaced with 2-bromobenzoic acid and 2, 3-dimethylaniline by the method described in example 1 above, and the feeds were dosed in proportion to give 0.82g of a yellow-green solid in 53.8% overall yield.¹H-NMR(400MHz,DMSO-D 6)δ:10.46(s,1H),8.23(dd,J＝8.1,1.3Hz,1H),8.09(d,J＝8.2Hz,1H),7.99(d,J＝8.4Hz,1H),7.76–7.69(m,1H),7.21(ddd,J＝8.0,7.0,0.9Hz,1H),7.09(d,J＝8.2Hz,1H),2.46(s,3H),2.41(s,3H).13C NMR(101MHz,DMSO-D 6)δ:173.75,138.46,137.53,136.68,127.47,121.32,120.64,120.31,120.08,119.31,118.09,117.36,116.01,18.36,10.65.MS(ESI):m/z 224.11[M+H]+.

EXAMPLE 62 preparation of methoxy-5, 6-dimethyl-9-acridone

The starting materials, 2-bromophenylacetic acid and 2-amino-3, 4-dimethylbenzoic acid, were replaced with 2-bromo-5-methoxybenzoic acid and 2, 3-dimethylaniline by the method described in example 1 above, and the materials were dosed in proportion to give 1.53g of a pale yellow solid with a total yield of 40.2%.¹H-NMR(400MHz,DMSO-D 6)δ:10.49(s,1H),8.13(d,J＝8.3Hz,1H),7.93(d,J＝9.1Hz,1H),7.62(d,J＝3.0Hz,1H),7.35(dd,J＝9.1,3.0Hz,1H),7.11(d,J＝8.3Hz,1H),3.88(s,3H),2.45(s,3H),2.36(s,3H).13C NMR(101MHz,DMSO-D 6)δ(ppm):13C NMR(101MHz,DMSO-D6)δ:176.69,154.43,141.59,139.66,136.48,124.17,123.76,123.55,123.04,121.02,120.35,119.23,105.21,55.74,21.16,13.67.MS(ESI):m/z 254.12[M+H]+。

EXAMPLE 72 preparation of methoxy-3, 4-dimethyl-9-acridone

The starting materials 2-bromobenzoic acid and 2-amino-3, 4-dimethylbenzoic acid were replaced by 2-bromobenzoic acid and 2, 3-dimethyl-4-methoxyaniline by the method described in example 1 above, and the batch was charged in proportion to give 0.31g of a dark green solid with a total yield of 57.3%. 1H-NMR (400MHz, DMSO-D6) δ:10.48(s,1H),8.21(D, J ═ 7.9Hz,1H),7.93(D, J ═ 8.4Hz,1H),7.66(t, J ═ 7.6Hz,1H),7.53(s,1H),7.26(t, J ═ 7.2Hz,1H),3.84(s,3H),2.55(s,3H),2.31(s,3H), 13C NMR (101MHz, DMSO-D6) δ:176.73,152.74,141.17,134.65,133.21,132.59,126.22,125.12,121.21,119.85,119.46,118.55,101.67,55.87,14.29,13.38.ms (esi): M/z 254.12[ M + H ] +.

EXAMPLE 82 preparation of 7, 7-dimethoxy-3, 4-dimethyl-9-acridone

The starting materials, 2-bromophenylacetic acid and 2-amino-3, 4-dimethylbenzoic acid, were replaced with 2-bromo-5-methoxybenzoic acid and 2, 3-dimethyl-4-methoxyaniline according to the procedure described in example 1 above, and the feeds were dosed in proportions to give 0.42g of a dark green solid with a total yield of 44.4%. 1H-NMR (400MHz, DMSO-D6) δ:10.45(s,1H),7.86(D, J ═ 9.1Hz,1H),7.56(D, J ═ 2.9Hz,1H),7.51(s,1H),7.34(dd, J ═ 9.1,3.0Hz,1H),3.89(s,3H),3.83(s,3H),2.54(s,3H),2.31(s,3H), 13C NMR (101MHz, DMSO-D6) δ:175.69,154.46,152.45,136.21,134.42,132.32,125.13,124.12,120.43,120.26,118.68,104.45,101.33,55.91,55.67,14.28,13.31.ms (esi): M/z 284.13[ M + H ] +.

The chemical names and structural formulae of the compounds prepared in examples 1-8 and the positive control drugs cGAMP, DMXAA and CMA are shown in the following table.

EXAMPLE 9 evaluation of agonism of STING pathway Activity by Compounds on a cellular model

9.1 test principle:

compounds were co-cultured with engineered cell lines, all derived from Invivogen: 293T-Dual^TM hSTING-R232 Cells；293T-Dual^TMmSTING Cells. The 293 human embryo kidney cell has no STING per se and is a good STING pathway verification cell. hSTING-R232 is commonly referred to as wild type STING and accounts for about 70% of the population. mSTING Cells transfected mSTING 293T Cells.

9.2. The test method comprises the following steps:

180 μ L of 293T-Dual^TMhSTING-R232 Cells or 293T-Dual^TMAfter culturing mSTING Cells (100000 Cells/well) together with 20. mu.L of saline solution or test compound saline solution in a 5% CO2 incubator at 37 ℃ for 48 hours, the activity of each pathway of STING was indirectly measured using different test solutions: 293T cells were assayed for IRF pathway activity using QUANTI-Blue and NF-. kappa.B pathway activity using QUANTI-Blue. The test was carried out in two steps, the first step of preliminary screening for effective compounds, each at a preliminary screening concentration of 10. mu.M and 100. mu.M, and the positive compound cGAMP at a concentration of 20. mu.M. The second step is to set concentration gradient for effective small molecules to determine the EC50 value of the compound and compare with cGAMP to judge the strength of the molecule activating the STING pathway.

9.3. And (3) testing results:

TABLE 1 biological rating test results of target Compounds agonizing the STING pathway

Compound numbering	Human STING Activity	Murine STING Activity
			Con 1	++	++
Con 2	-	++
			Con 3	-	++
3	+	+
			6	++	++

TABLE 2 biological Activity test results of target Compounds for agonizing STING pathway

Note: in tables 1 and 2, (1) — "indicates no activity; "+" indicates good activity; "+ +" indicates excellent activity; (2) EC 50: half maximal effect concentration of drug; emax: maximum intensity value of drug effect; nt indicates no activity detected.

EXAMPLE 10 evaluation of Competition binding of Compounds with STING ligand

10.1. The test principle is as follows:

the detection principle is based on

The technique is that when a dye approaches, excitation of a donor by a light source (laser or flash lamp) triggers Fluorescence Resonance Energy Transfer (FRET) to an acceptor, which fluoresces at a specific wavelength (665 nm). Specific anti-6 His antibodies were labeled with cryptate terbium (donor) and bound to a 6 His-labeled human STING protein and a STING ligand labeled d2 (receptor). Human STING ligands are detected in a competitive assay, the detection compound competes with the tail dominator labeled d2, thereby blocking FRET, and the magnitude of the specific signal is inversely proportional to the concentration of the compound.

10.2. The test method comprises the following steps:

adding 5 μ L of detection compound or cGAMP or negative control solution with different concentrations, adding 5 μ L of 6 His-labeled human STING (detection buffer solution is added into negative control hole), and adding 10 μ L of d2 ligand and anti-6 His antibody mixture; after the cell plate is sealed and cultured for 3h at room temperature, the absorbance values at 665nm and 620nm are respectively read by an enzyme-labeling instrument, and the ratio AU665/AU620 of the absorbance values is the basis for evaluating the strength of the compound combined with STING protein.

10.3. And (3) testing results:

TABLE 3 competitive inhibition of STING binding assay results for Compounds

Compound numbering	Competitive inhibitory concentration
		3	1721.54±15.68nM
6	375.25±7.65nM
		Con1	333.32±6.89nM
Con2	>500μM

Experiments prove that the compound has agonists on different species, particularly human STING, and has high activity.

The foregoing is illustrative of the present invention and is not to be construed as limiting thereof. The practice of the present invention will employ, unless otherwise indicated, conventional techniques of organic chemistry, polymer chemistry, biotechnology and the like, and it will be apparent that the invention may be practiced otherwise than as specifically described in the foregoing description and examples. Other aspects and modifications within the scope of the invention will be apparent to those skilled in the art to which the invention pertains. Many modifications and variations are possible in light of the above teaching and are therefore within the scope of the invention.

Claims

1. A compound of formula I, a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof,

wherein the content of the first and second substances,

2. The compound according to claim 1, a pharmaceutically acceptable salt, solvate, hydrate or prodrug compound thereof, wherein the compound is selected from the group consisting of:

9, 10-dimethyl-6H-pyrrolo [3,2,1-de ] -1, 6-acridinedione,

2- (5, 6-dimethyl-9-acridone) -4-acetic acid,

2- (5, 6-dimethyl-7-methoxy-9-acridone) -4-acetic acid,

2-methoxy-5, 6-dimethyl-9-acridone-4-carboxylic acid,

3, 4-dimethyl-9-acridone,

2-methoxy-5, 6-dimethyl-9-acridone,

2-methoxy-3, 4-dimethyl-9-acridone, and

2, 7-dimethoxy-3, 4-dimethyl-9-acridone.

3. A pharmaceutical composition comprising a compound of claim 1, a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof, and one or more pharmaceutically acceptable carriers or excipients.

4. The pharmaceutical composition according to claim 3, for use in the treatment and/or prevention of a STING pathway related disease.

5. The use of a compound of claim 1, a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, for the manufacture of a medicament for the treatment and/or prevention of a disease associated with the STING pathway.

6. The use according to claim 5, wherein the diseases associated with the STING pathway are infectious diseases, cancer, inflammation, allergy and autoimmune diseases.

7. The use according to claim 6, wherein the infectious disease is a viral infectious disease or a bacterial infectious disease, the etiological agent of which is a viral pathogen such as influenza virus, enterovirus, hepatitis B virus, hepatitis C virus, Ebola virus, Marburg virus, SARS virus, Securia virus, bunyavirus, rhinovirus, respiratory nucleopolyvirus, cholera virus, or the like, or a bacterial pathogen such as Mycobacterium tuberculosis, Escherichia coli, Acinetobacter baumannii, Diplococcus pneumoniae, Streptococcus lactis, Sarcina urensis, Staphylococcus aureus, Bacillus subtilis, Bacillus anthracis, Bacillus subtilis, Bacillus streptococci, Bacillus proteus, Vibrio cholerae, Treponema pallidum, or the like.

8. The use according to claim 6, wherein the cancer is gastric cancer, liver cancer, lung cancer, esophageal cancer, cervical cancer, breast cancer, colon cancer, rectal cancer, nasopharyngeal cancer, ovarian cancer, renal cancer, bladder cancer, thyroid cancer, skin cancer, or the like; malignant tumors derived from mesenchymal tissues such as muscle, fat, bone, blood vessel, lymph, etc., for example, rhabdomyosarcoma, leiomyosarcoma, fibrosarcoma, liposarcoma, osteosarcoma, chondrosarcoma, angiosarcoma, lymphosarcoma, etc. Also, for example, leukemia, Hodgkin's disease, Wilms ' tumor (nephroblastoma), melanoma, retinoblastoma, seminoma, granuloma, Kupffer's tumor, Ewing's tumor, malignant vascular endothelial cell tumor, or Paget's disease of the breast.

9. Use according to claim 6, wherein the inflammatory, allergic and autoimmune diseases are musculoskeletal, vascular, neurological, digestive, ocular, reproductive or other inflammatory diseases, such as asthma.

10. A process for the preparation of a compound according to claim 1, comprising the steps of: