A class of urea derivatives containing benzoheterocyclic substitutions and their preparation and use

Abstract

The present invention relates to urea derivatives containing benzoheterocyclic substitutions and their preparation and use. Specifically, the compound of the present invention has a structure shown in Formula I, wherein the definitions of each group and substituent are as described in the specification. The present invention also discloses a preparation method of the compound and its use in treating autoimmune diseases.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a class of urea derivatives containing benzoheterocyclic substitutions and preparation and use thereof. Background Art

[0002] Autoimmune diseases (ADs) have a high incidence rate (5%-8%) worldwide and are of various types. Currently, more than 80 autoimmune diseases have been identified, including systemic lupus erythematosus, type I diabetes, rheumatoid arthritis, inflammatory bowel disease, etc. Most patients need to take medication for a long time or even for life, which seriously affects the quality of life of patients and even threatens their lives. Therefore, autoimmune diseases have become the third largest killer threatening human health after cardiovascular diseases and cancer.

[0003] Currently, immunomodulatory drugs commonly used to treat autoimmune diseases (such as azathioprine, methotrexate, glucocorticoids, cyclosporine A, etc.) have a wide range of effects, are non-disease specific, and are associated with side effects such as infections and malignant tumors. Drugs used to block various pathways and components of the immune system, such as cytokines, cell adhesion molecules, and co-stimulatory molecules, are all biological macromolecules with high treatment costs and more serious side effects. Therefore, exploring autoimmune disease-specific small molecule drugs with new mechanisms is the most urgent need facing this field.

[0004] The stimulator of interferon genes (STING) is an important signaling molecule in the innate immune cGAS-STING pathway. When cGAS senses endogenous or exogenous double-stranded DNA (dsDNA), it is activated. The cyclic guanosine monophosphate (cGAMP) generated by its catalysis activates STING, which then recruits TBK1 protein and phosphorylates interferon regulatory factor 3 (IRF3) to activate the innate immune system. When the dsDNA that activates the STING signal is derived from necrosis or inappropriate apoptosis, it will promote the secretion of inflammatory cytokines and cause inflammation and autoimmune diseases. Existing research results have shown that negative regulation of the STING signaling pathway is expected to become an important strategy for the treatment of autoimmune diseases.

[0005] Although many research results have verified that inhibiting the cGAS-STING pathway is an important strategy for treating autoimmune diseases, research on inhibitors targeting the cGAS-STING pathway is still in its early stages. In 2018, Simone M. Haag et al. reported a class of nitrofuran and indole compounds C-178 (Formula II) and H-151 (Formula III) in Nature. These compounds can covalently bind to Cys91 of the STING protein, blocking palmitoylation induced by STING activation, thereby blocking its assembly into a multimeric complex in the Golgi apparatus and inhibiting downstream signaling pathway conduction. In addition, this type of inhibitor can reduce the production of inflammatory cytokines mediated by STING protein in human and mouse cells and reduce the pathological characteristics of autoinflammatory diseases in mice. In the same year, Wang Chen et al. reported the small molecule AstinC (Formula IV) isolated from Aster tataricus in Cell Reports. Astin C can competitively bind to cyclic dinucleotides (CDNs) and inhibit STING (STING-R232 Kd = 53nM). In the Trex1 gene-deficient mouse model, intravenous injection of Astin C (1 mg / kg) can reduce the expression of cytokines and the abundance of autoantibodies in serum, significantly alleviating the autoinflammatory response of mice. In addition, in 2019, Merck reported a class of STING antagonists with a tetrahydroisoquinoline core structure in ACS Med. Chem. Lett. This class of compounds can competitively bind to the cGAMP binding site, but the cell activity is weak.

[0006]

[0007] Based on the therapeutic potential of STING inhibitors for autoimmune diseases, major pharmaceutical giants have focused on the development of STING inhibitors. In September 2018, Novartis signed an $840 million cooperation agreement with IFM Due to jointly conduct research on cGAS-STING inhibitors. However, the structural types of STING inhibitors reported so far are extremely limited, and their activity is mostly at the molecular level. The cellular activity is mostly weak, and no inhibitors have entered clinical research.

[0008] Therefore, it is urgent to conduct research on a STING inhibitor with excellent cellular and in vivo activity and strong stability. Summary of the invention

[0009] The object of the present invention is to provide a compound represented by formula I, a preparation method thereof and an anti-tumor use thereof.

[0010] In a first aspect of the present invention, there is provided a compound of formula I, or an isomer, prodrug, solvate, hydrate or a pharmaceutically acceptable salt thereof,

[0011]

[0012] in,

[0013] A is a 5- to 18-membered ring, which includes 1-4 heteroatoms; the heteroatoms are selected from oxygen, nitrogen, sulfur, and boron;

[0014] X is selected from CR1, N;

[0015] n is an integer selected from 0 to 4, such as 0, 1, 2, 3, 4;

[0016] R1 is selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, R a Substituted or unsubstituted C1-C8 alkyl, R a Substituted or unsubstituted C2-C8 alkenyl, R a Substituted or unsubstituted C2-C8 alkynyl, R a Substituted or unsubstituted C1-C8 alkoxy, R a Substituted or unsubstituted C1-C8 alkyl acyl, R a Substituted or unsubstituted aminoacyl, R a Substituted or unsubstituted C1-C8 alkylamide, R a Substituted or unsubstituted C1-C8 alkenyl amide, R a Substituted or unsubstituted C1-C8 alkylamino, R a Substituted or unsubstituted C1-C8 alkyl sulfide group, R a Substituted or unsubstituted 3-6 membered heterocyclic group, R a Substituted or unsubstituted 3-6 membered cycloalkyl, R a Substituted or unsubstituted 5-10 membered aryl, R a Substituted or unsubstituted 5-10 membered heteroaryl;

[0017] R2 is selected from hydrogen, halogen, R a Substituted or unsubstituted C1-C8 alkyl, R a Substituted or unsubstituted C1-C8 alkylamino;

[0018] m is an integer selected from 0 to 6, such as 0, 1, 2, 3, 4, 5, 6;

[0019] R3 are the same or different and are independently selected from hydrogen, halogen, hydroxyl, R a Substituted or unsubstituted C1-C8 alkyl;

[0020] R aSelected from hydrogen, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C1-C8 haloalkyl, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted C1-C8 alkoxyacyl, substituted or unsubstituted C1-C8 hydroxyalkyl, substituted or unsubstituted C1-C8 alkylamino, substituted or unsubstituted 3-6 membered heterocyclyl, substituted or unsubstituted 3-6 membered cycloalkyl, substituted or unsubstituted 5-6 membered heteroaryl, phosphate, wherein the substitution is selected from hydrogen, halogen, hydroxyl, cyano, C1-C8 alkyl, C1-C8 alkylacyl, 3-6 membered heterocyclyl, 3-6 membered cycloalkyl, C1-C4 alkoxy.

[0021] In another preferred embodiment, A is a saturated or unsaturated or aromatic ring.

[0022] In another preferred embodiment, A is a 5-12 (eg, 5, 6, 7, 8, 10, 12) membered ring including 1-4 (eg, 1, 2, 3, 4) heteroatoms selected from oxygen, nitrogen, sulfur and boron.

[0023] In another preferred embodiment, A is a 5-6 membered ring, which includes at least 2 heteroatoms selected from oxygen and boron.

[0024] In another preferred embodiment, A is a 5-6 membered ring which includes 2 heteroatoms selected from oxygen and boron.

[0025] In another preferred embodiment, A is selected from dioxolane, dioxane, borane, and 12-crown-4 ether.

[0026] In another preferred embodiment, A is selected from

[0027] In another preferred embodiment, R1 is selected from hydrogen, halogen, R a Substituted or unsubstituted C1-C6 alkyl, R a Substituted or unsubstituted C2-C6 alkenyl, R a Substituted or unsubstituted C2-C6 alkynyl, R a Substituted or unsubstituted C1-C6 alkoxy, R a Substituted or unsubstituted C1-C6 alkyl sulfide group, R a Substituted or unsubstituted C1-C6 aminoacyl, R a Substituted or unsubstituted C1-C6 alkylamide, R a Substituted or unsubstituted 3-6 membered heterocyclic group, R a Substituted or unsubstituted 5-6 membered heteroaryl, wherein R a As mentioned above.

[0028] In another preferred embodiment, R1 is selected from hydrogen, halogen, R a Substituted or unsubstituted C1-C6 alkyl, Ra Substituted or unsubstituted C2-C6 alkenyl, R a Substituted or unsubstituted C2-C6 alkynyl, R a Substituted or unsubstituted C1-C6 alkoxy, R a Substituted or unsubstituted C1-C6 alkyl sulfide group, R a Substituted or unsubstituted C1-C6 aminoacyl, R a Substituted or unsubstituted 5-6 membered heteroaryl, wherein R a As mentioned above.

[0029] In another preferred embodiment, R2 is selected from hydrogen, R a Substituted or unsubstituted C1-C8 alkyl, wherein R a As mentioned above, hydrogen is preferred.

[0030] In another preferred embodiment, R3 are the same or different, preferably the same.

[0031] In another preferred embodiment, R3 is independently selected from hydrogen, halogen, hydroxyl, R a Substituted or unsubstituted C1-C8 alkyl, wherein R a As mentioned above.

[0032] In another preferred embodiment, R a Selected from hydrogen, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C1-C6 haloalkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkoxyacyl, substituted or unsubstituted C1-C6 alkylhydroxy, substituted or unsubstituted 3-6 membered cycloalkyl, substituted or unsubstituted 3-6 membered heterocyclyl, substituted or unsubstituted 5-6 membered heteroaryl, phosphate, the substitution is selected from hydrogen, halogen, hydroxyl, cyano, C1-C8 alkyl, C1-C8 alkanoyl, 3-6 membered heterocyclyl, 3-6 membered cycloalkyl, C1-C4 alkoxy.

[0033] In another preferred embodiment, ring A is a 5- to 18-membered ring, which includes 1-4 heteroatoms; the heteroatoms are selected from oxygen, nitrogen, sulfur and boron.

[0034] X is selected from CH, N;

[0035] R1 is selected from hydrogen, halogen, hydroxyl, amino, cyano, nitro, Ra substituted or unsubstituted C1-C8 alkyl, Ra substituted or unsubstituted C2-C8 alkenyl, Ra substituted or unsubstituted C2-C8 alkynyl, Ra substituted or unsubstituted C1-C8 alkoxy, Ra substituted or unsubstituted C1-C8 alkylacyl, Ra substituted or unsubstituted aminoacyl, Ra substituted or unsubstituted C1-C8 alkylamide, Ra substituted or unsubstituted C1-C8 alkenylamide, Ra substituted or unsubstituted C1-C8 alkylamino, Ra substituted or unsubstituted C1-C8 alkylthioether, Ra substituted or unsubstituted 3-6 membered heterocyclyl, Ra substituted or unsubstituted 3-6 membered cycloalkyl, Ra substituted or unsubstituted 5-6 membered heteroaryl;

[0036] R2 is selected from hydrogen, halogen, Ra substituted or unsubstituted C1-C8 alkyl, Ra substituted or unsubstituted C1-C8 alkylamino;

[0037] m is an integer selected from 0 to 6;

[0038] R3 is independently selected at each occurrence from hydrogen, halogen, hydroxy, Ra substituted or unsubstituted C1-C8 alkyl;

[0039] Ra is selected from hydrogen, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted C1-C8 alkoxyacyl, substituted or unsubstituted C1-C8 alkylamino, substituted or unsubstituted 3-6 membered heterocyclyl, substituted or unsubstituted 3-6 membered cycloalkyl, substituted or unsubstituted 5-6 membered heteroaryl, phosphate, and the substitution is selected from hydrogen, halogen, hydroxyl, cyano, C1-C8 alkyl, C1-C8 alkanoyl, 3-6 membered heterocyclyl, 3-6 membered cycloalkyl, C1-C4 alkoxyalkoxy.

[0040] In another preferred embodiment, the compound is selected from the compounds listed in Table 1.

[0041] Table 1

[0042]

[0043]

[0044]

[0045] In the second aspect of the present invention, there is provided a method for preparing a compound of formula I, or an isomer, prodrug, solvate, hydrate or a pharmaceutically acceptable salt thereof, the method being selected from the group consisting of:

[0046] Method 1, comprising the steps of:

[0047]

[0048] a-1) reacting the compound of formula 1 in the presence of an azidating agent to obtain a compound of formula 2;

[0049]

[0050] a-2) the compound of formula 2 is subjected to rearrangement reaction to obtain a compound of formula 3;

[0051]

[0052] a-3) reacting a compound of formula 3 with a compound of formula 4 to obtain a compound of formula I;

[0053] Wherein, A, R1, R2, R3, and m are as defined in the first aspect of the present invention;

[0054] Method 2, including the steps:

[0055]

[0056] b) reacting the compound of formula 1 with the compound of formula 4 in the presence of microwaves and an azo reagent to obtain a compound of formula I;

[0057] Wherein, A, R1, R2, R3, m and n are as defined in the first aspect of the present invention.

[0058] In another preferred embodiment, the azotizing agent is selected from the group consisting of diphenylphosphoryl azide, sodium azide, trimethylsilyl azide, tributyltin azide, tetrabutylammonium azide, or a combination thereof.

[0059] In another preferred embodiment, step a-2) comprises: heating the compound of formula 2 under reflux to obtain the compound of formula 3.

[0060] In another preferred embodiment, the power of the microwave is 100-200W.

[0061] In another preferred embodiment, the method is selected from the following group:

[0062] Method 1, including the steps:

[0063]

[0064] Method 2, including the steps:

[0065]

[0066] In a third aspect of the present invention, a pharmaceutical composition is provided, comprising:

[0067] (i) one or more therapeutically effective amounts of the compounds of the first aspect of the present invention, or their isomers, prodrugs, solvates, hydrates or pharmaceutically acceptable salts; and

[0068] (ii) a pharmaceutically acceptable carrier.

[0069] In another preferred embodiment, the pharmaceutical composition is an injection, capsule, tablet, pill, powder or granule.

[0070] In another preferred embodiment, the pharmaceutical composition further comprises one or more second therapeutic agents, and the second therapeutic agent is a drug for preventing and / or treating autoimmune diseases.

[0071] In the fourth aspect of the present invention, there is provided a use of the compound described in the first aspect of the present invention, or its isomer, prodrug, solvate, hydrate or pharmaceutically acceptable salt, or the pharmaceutical composition described in the third aspect of the present invention, for preparing a preparation for preventing and / or treating diseases associated with interferon gene stimulator protein.

[0072] In another preferred embodiment, the disease associated with interferon gene stimulated protein is selected from the following group: Singleton-Merten syndrome (SMS), Aicardi-Goutières syndrome (AGS), systemic lupus erythematosus (SLE), familial pernio lupus erythematosus (FCL), retinal vasculopathy and leukodystrophy (RVCL), STING-associated vasculopathy of infancy (SAVI), psoriasis, scleroderma, stroke, myocardial infarction, brain trauma, atherosclerosis-related vascular disease, cardiovascular disease, diabetes and its complications, Parkinson's disease, Huntington's disease, familial amyotrophic lateral sclerosis, neurodegenerative diseases, small intestinal malabsorption syndrome, irritable bowel syndrome, Sjogren's syndrome, multiple sclerosis, Crohn's disease, non-alcoholic fatty hepatitis, rheumatoid arthritis, inflammatory bowel disease, ulcerative colitis, autoimmune colitis, suppurative hidradenitis, uveitis, mucositis and other inflammatory diseases and autoimmune diseases.

[0073] In the fifth aspect of the present invention, an interferon gene stimulator protein inhibitor is provided, which includes one or more compounds described in the first aspect of the present invention, or their isomers, prodrugs, solvates, hydrates or pharmaceutically acceptable salts.

[0074] In the sixth aspect of the present invention, a method for preventing and / or treating diseases associated with interferon gene stimulator protein is provided, comprising the steps of administering a therapeutically effective amount of one or more compounds described in the first aspect of the present invention, or their isomers, prodrugs, solvates, hydrates or pharmaceutically acceptable salts thereof, or the pharmaceutical composition described in the third aspect of the present invention to a patient in need.

[0075] In another preferred embodiment, the method is diagnostic or non-diagnostic.

[0076] In another preferred embodiment, the method is therapeutic or non-therapeutic.

[0077] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0078] After long and in-depth research, the inventor unexpectedly prepared a compound shown in formula I with simple structure, convenient synthesis and stable metabolism, which has excellent inhibitory effect on interferon gene stimulating protein, thereby inhibiting the conduction of downstream signal pathways, thereby achieving effective treatment of inflammatory and autoimmune diseases. On this basis, the inventor completed the present invention.

[0079] the term

[0080] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0081] As used herein, when used in reference to a specific recited numerical value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0082] As used herein, the term "comprising" or "including (comprising)" may be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

[0083] In the present invention, the halogen is F, Cl, Br or I.

[0084] In the present invention, the term "C1-C6" refers to having 1, 2, 3, 4, 5 or 6 carbon atoms, "C1-C8" refers to having 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms, and so on. "5-6 membered" refers to having 5-6 ring atoms, and so on.

[0085] In the present invention, the term "alkyl" refers to a saturated linear or branched hydrocarbon moiety. For example, the term "C1-C8 alkyl" refers to a straight or branched alkyl group having 1 to 8 carbon atoms, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl, etc.; preferably ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl.

[0086] In the present invention, the term "haloalkyl" refers to the above-mentioned alkyl group substituted by a halogen atom, including fluoroalkyl, chloroalkyl, bromoalkyl or iodoalkyl, including but not limited to trifluoromethyl, pentafluoroethyl, heptafluoroisopropyl, monochloromethyl, dichloromethyl, trichloromethyl and the like.

[0087] In the present invention, the term "alkoxy" refers to an -O-(C1-C8 alkyl) group. For example, the term "C1-C8 alkoxy" refers to a straight or branched alkoxy group having 1 to 8 carbon atoms, including, but not limited to, methoxy, ethoxy, propoxy, isopropoxy, and butoxy.

[0088] In the present invention, the term "alkyl sulfide group" means -S-(C1-C8 alkyl) group. For example, the term "C1-C8 alkyl sulfide group" refers to a straight or branched alkyl sulfide group having 1 to 8 carbon atoms, including but not limited to methyl sulfide group (-S-CH3), ethyl sulfide group (-S-CH2CH3), propyl sulfide group, isopropyl sulfide group and butyl sulfide group.

[0089] In the present invention, the term "alkenyl" refers to a straight chain or branched hydrocarbon moiety containing at least one double bond. For example, the term "C2-C8 alkenyl" refers to a straight chain or branched alkenyl group containing one double bond having 2 to 8 carbon atoms, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl.

[0090] In the present invention, the term "alkynyl" refers to a straight or branched alkynyl group containing one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl.

[0091] In the present invention, the term "cycloalkyl" refers to a saturated cyclic hydrocarbon moiety, for example, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3 to 8 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and cyclodecyl, etc. The terms "C3-C6 cycloalkyl" and "C5-C6 cycloalkyl" have similar meanings.

[0092] In the present invention, the term "heterocyclyl" refers to a cyclic group containing at least one (e.g., 1, 2, 3) intracyclic heteroatom (e.g., N, O or S), such as tetrahydrofuranyl, pyrrolyl, tetrahydropyridyl or pyrrolidinyl, dioxopentanyl, dioxhexatanyl, oxaziridine and oxaziridine.

[0093] In the present invention, the term "aryl" refers to a hydrocarbon moiety containing one or more aromatic rings. For example, the term "C6-C10 aryl" refers to an aromatic ring group having 6 to 10 carbon atoms without heteroatoms in the ring, such as phenyl, naphthyl, etc.

[0094] In the present invention, the term "heteroaryl" refers to an aromatic cyclic group containing at least one (e.g., 1, 2, 3) intracyclic heteroatom (e.g., N, O or S), such as furanyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, triazolyl, tetrazolyl, etc.

[0095] The term "alkylacyl" means a -CO-alkyl group as defined above.

[0096] The term "alkylamido" refers to an -NH-CO-alkyl group as defined above.

[0097] The term "alkenylamido" refers to an alkenyl group as defined above -NH-CO-.

[0098] The term "alkylamino" refers to -NH-alkyl as defined above and -N-alkyl as defined above.

[0099] The term "hydroxyalkyl" means an -alkyl-OH group as defined above.

[0100] The term "alkoxyacyl" means a -CO-alkoxy group as defined above.

[0101] The term "phosphate" means For example, PO(OEt)2, PO(OMe)2.

[0102] Unless otherwise specified, the alkyl, haloalkyl, alkoxy, alkylthioether, cycloalkyl, heterocyclic and aryl groups described herein are substituted and unsubstituted groups. Possible substituents on alkyl, haloalkyl, alkoxy, alkylthioether, cycloalkyl, heterocyclic and aryl groups include, but are not limited to: hydroxy, amino, nitro, nitrile, halogen, C1-C6 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C20 cycloalkyl, C3-C20 cycloalkenyl, C1-C20 heterocycloalkyl, C1-C20 heterocycloalkenyl, C1-C6 alkoxy, aryl, heteroaryl, heteroaryloxy, C1-C10 ... The cycloalkyl group, heterocycloalkyl group, heterocycloalkenyl group, aryl group, and heteroaryl group may be condensed with each other.

[0103] In the present invention, the substitution is mono- or poly-substitution, and the poly-substitution is di-, tri-, tetra- or penta-substitution. The di-substitution means having two substituents, and so on.

[0104] Salt type

[0105] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

[0106] Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0107] The term "solvate" refers to a complex formed by the coordination of the compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the coordination of the compound of the present invention with water.

[0108] The term "prodrug" includes a substance which may be biologically active or inactive, and which undergoes metabolism or chemical reactions in the human body after being taken by an appropriate method to be converted into a compound of Formula I, or a salt or solution composed of a compound of Formula I. The prodrug includes (but is not limited to) carboxylate, carbonate, phosphate, nitrate, sulfate, sulfone, sulfoxide, amino compound, carbamate, azo compound, phosphoramide, glucoside, ether, acetal and the like forms of the compound.

[0109] Pharmaceutical compositions and methods of administration

[0110] The present invention also provides a pharmaceutical composition comprising:

[0111] (i) one or more therapeutically effective amounts of a compound of Formula I, or an isomer, prodrug, solvate, hydrate, or a pharmaceutically acceptable salt thereof; and

[0112] (ii) a pharmaceutically acceptable carrier.

[0113] Since the compounds of the present invention have excellent anti-tumor activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate tumor-related diseases.

[0114] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. The therapeutically effective amount is determined based on the age, condition, course of treatment, etc. of the subject. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention / dose, and more preferably, contains 10-1000 mg of the compound of the present invention / dose. Preferably, the "one dose" is a capsule or tablet.

[0115] The "pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include sugars (such as glucose, sucrose, lactose, etc.), starches (such as corn starch, potato starch, etc.), cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0116] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0117] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0118] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0119] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0120] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0121] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0122] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0123] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0124] The aminobenzimidazole derivatives represented by the above formula I and pharmaceutically acceptable salts thereof in the present invention can be administered alone or in combination with other pharmaceutically acceptable therapeutic agents.

[0125] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

[0126] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0127] The main advantages of the present invention include:

[0128] (1) The compound has the characteristics of simple structure, easy synthesis and diverse administration methods;

[0129] (2) The compound has an excellent inhibitory effect on interferon gene-stimulating protein, thereby achieving effective treatment of autoimmune diseases;

[0130] (3) The half inhibitory concentration (IC) of the compound on the activation of interferon gene stimulator protein in human cells 50 ) can be less than 20nM.

[0131] The present invention will be further described below in conjunction with specific implementation. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0132] For the following examples, standard operations and purification methods known to those skilled in the art can be used. Unless otherwise specified, raw materials are generally available from commercial sources, such as Aldrich Chemicals Co. and AcrosOrganics. Commercially available solvents and reagents are generally used without further purification, anhydrous solvents are treated by standard methods, and other reagents are commercially available analytically pure. Unless otherwise specified, all temperatures are expressed in ° C (degrees Celsius), and room temperature or ambient temperature refers to 20-25 ° C. The structure of the compound is determined by nuclear magnetic resonance spectroscopy (NMR). The nuclear magnetic resonance hydrogen spectrum shift (δ) is given in parts per million (ppm). The nuclear magnetic resonance hydrogen spectrum was measured using a Mercury-400MHz nuclear magnetic resonance instrument, deuterated chloroform (CDCl3) and deuterated methanol (CD3OD) were solvents, and tetramethylsilane (TMS) was an internal standard.

[0133] Chromatographic columns generally use 200-300 mesh silica gel as a carrier.

[0134] Example 1. Preparation of compounds

[0135] The following preparation examples exemplify the preparation of some compounds of formula I of the present invention, and each compound is represented by S1 to S27.

[0136] 1. Synthesis of Compound S1

[0137]

[0138] Step 1: Compound 1a (1 eq) was dissolved in toluene, triethylamine (1 eq) was added under nitrogen protection, and diphenylphosphoryl azide (1.1 eq) was added after about 15 minutes, and stirred at room temperature overnight. After the reaction was complete, the solvent was dried by spin drying, and then purified by column to obtain compound 1b. 1 H NMR (400MHz, CDCl3) δ8.70 (s, 1H), 8.31–8.22 (m, 1H), 7.95 (d, J = 2.8Hz, 1H), 7.46–7.41 (m, 1H), 7.34–7.29 (m, 2H).

[0139]

[0140] Step 2: Compound 1b was dissolved in toluene and refluxed overnight under nitrogen protection. After the reaction was complete, the solvent was dried by spin drying and pumping to obtain compound 1c. 1H NMR (400MHz, CDCl3) δ7.91 (s, 1H), 7.62 (d, J = 7.9Hz, 1H), 7.36 (d, J = 8.2Hz, 1H), 7.29–7.25 (m, 2H), 7.20 (t, J = 7.5Hz, 1H), 7.05 (d, J = 2.3Hz, 1H).

[0141]

[0142] Step 3: Compound 1c (1 eq) was dissolved in N,N-dimethylformamide, and compound 1d (1.2 eq) and triethylamine (3 eq) were added, and the mixture was reacted overnight at room temperature. After the reaction was complete, the reaction solution was poured into water, extracted with ethyl acetate, and purified by column to obtain compound S1. 1 H NMR (400MHz, MeOD) δ7.59(d,J=1.9Hz,1H),7.52(d,J=7.9Hz,1H),7.42(s,1H),7.35(d,J=8.2Hz,1H ),7.13(t,J=7.3Hz,1H),7.10(d,J=8.7Hz,1H),7.04(t,J=7.4Hz,1H),7.00(dd,J=8.7,2.1Hz,1H).

[0143] 2. Synthesis of Compound S2

[0144]

[0145] Compound 1a (1 eq), compound 2a (1.5 eq), diphenylphosphoryl azide (1.2 eq) and triethylamine (3 eq) were mixed in toluene and reacted for 15 minutes using a CEM Discover microwave synthesizer at 150 W and 100° C. After the reaction solution was cooled, it was poured into a saturated sodium bicarbonate solution, extracted with ethyl acetate, the organic phase was collected, and then purified by column to obtain compound S2. 1 H NMR (400MHz, MeOD) δ7.75(s,1H),7.54(d,J=7.9Hz,1H),7.46(s,1H),7.35(d,J=8.2Hz,1H),7.16–7.03(m,3H),6.86(d,J=8.1Hz,1H).

[0146] 3. Synthesis of Compound S3

[0147]

[0148] Compound 3a (1 eq), compound 1d (1.5 eq), diphenylphosphoryl azide (1.2 eq) and triethylamine (3 eq) were mixed in toluene and reacted for 15 minutes using a CEM Discover microwave synthesizer at 150 W and 100° C. After the reaction solution was cooled, it was poured into a saturated sodium bicarbonate solution, extracted with ethyl acetate, the organic phase was collected, and then purified by column to obtain compound S3. 1 H NMR(400MHz,DMSO)δ10.91(s,1H),8.78(s,1H),8.50(s,1H),7.71(s,1H),7.55(s,1H), 7.36–7.30(m,2H),7.23(d,J=9.3Hz,1H),7.11(d,J=8.1Hz,1H),6.95(t,J=9.3Hz,1H).

[0149] 4. Synthesis of Compound S4

[0150]

[0151] The synthesis method refers to compound S3, compound 4a is used to replace compound 3a, and the other experimental steps are the same to obtain compound S4. 1 H NMR (400MHz, DMSO) δ11.00(s,1H),8.75(s,1H),8.59(s,1H),7.70(d,J=2.0Hz,1H),7 .54(d,J=2.1Hz,2H),7.36(d,J=8.6Hz,1H),7.31(d,J=8.7Hz,1H),7.12-7.08(m,2H).

[0152] 5. Synthesis of Compound S5

[0153]

[0154] The synthesis method refers to compound S3, replacing compound 3a with compound 5a, and the other experimental steps are the same to obtain compound S5. 1 H NMR (400MHz, DMSO) δ11.01(s,1H),8.72(s,1H),8.60(s,1H),7.70–7.69(m,2H),7.52(d,J=2.4Hz,1H) ,7.32(d,J=5.0Hz,1H),7.30(d,J=5.2Hz,1H,7.20(dd,J=8.6,1.8Hz,1H),7.11(dd,J=8.8,2.1Hz,1H).

[0155] 6. Synthesis of Compound S6

[0156]

[0157] The synthesis method refers to compound S3, replacing compound 3a with compound 6a, and the other experimental steps are the same to obtain compound S6. 1 H NMR (400MHz, DMSO) δ11.56(s,1H),8.85(s,1H),8.59(s,1H),8.22(s,1H),7.75(dd,J=9.4,2.5Hz,1 H),7.69(d,J=1.9Hz,1H),7.63(d,J=2.4Hz,1H),7.31(d,J=8.7Hz,1H),7.13(dd,J=8.8,2.0Hz,1H).

[0158] 7. Synthesis of Compound S7

[0159]

[0160] The synthesis method refers to compound S3, replacing compound 1d with compound 7a, and the other experimental steps are the same to obtain compound S7. 1 H NMR (400MHz, DMSO) δ10.86(s,1H),8.43(s,1H),8.35(s,1H),7.53(d,J=2.3Hz,1H),7.33(dd,J=8.8,4.5Hz,1H),7.24(d,J= 1.9Hz,1H),7.21(dd,J=9.9,2.4Hz,1H),6.96-6.91(m,1H),6.82(d,J=8.3Hz,1H),6.77(dd,J=8.4,2.0Hz,1H),5.96(s,2H).

[0161] 8. Synthesis of Compound S8

[0162]

[0163] The synthesis method refers to compound S3, replacing compound 1d with compound 8a, and the other experimental steps are the same to obtain compound S8. 1 H NMR (400MHz, DMSO) δ10.93(s,1H),8.88(s,1H),8.56(s,1H),7.71(d,J=2.3Hz,1H),7.55(d,J=2.4Hz ,1H),7.39–7.33(m,2H),7.27(dd,J=9.0,2.4Hz,1H),7.22(dd,J=9.8,2.3Hz,1H),6.97–6.92(m,1H).

[0164] 9. Synthesis of Compound S9

[0165]

[0166] Step 1: Dissolve compound 9a in nitromethane, add a 1:1 mixed solution of concentrated sulfuric acid and concentrated nitric acid dropwise under an ice bath, and react for about half an hour. After the reaction is complete, slowly pour the reaction solution into water, extract with ethyl acetate, collect the organic phase, and purify it through a column to obtain compound 9b.

[0167]

[0168] Step 2: Dissolve compound 9b (1 eq) in methanol, add ammonium chloride (10 eq), heat to 80°C, and then add iron powder (5 eq), react for about 8 hours, filter while hot after the reaction is complete, wash the filter cake with methanol, collect the filtrate and spin-dry to obtain crude compound 9c.

[0169]

[0170] The synthesis method refers to compound S3, replacing compound 1d with compound 9c, and the other experimental steps are the same to obtain compound S9. 1 H NMR (400MHz, DMSO) δ10.87(s,1H),8.41(s,1H),8.37(s,1H),7.55(d,J=2.4Hz,1H),7.34(dd,J=8.9,4.5Hz,1H),7.28(d,J=2.1 Hz,1H),7.21(dd,J=9.8,2.4Hz,1H),6.97–6.90(m,3H),4.08–4.03(m,4H),3.76–3.74(m,2H),3.69–3.67(m,2H),3.64(s,4H).

[0171] 10. Synthesis of Compound S10

[0172]

[0173] The synthesis method refers to compound S3, replacing compound 1d with compound 10a, and the other experimental steps are the same to obtain compound S10. 1H NMR (400MHz, DMSO) δ10.85(s,1H),8.33(s,1H),8.31(s,1H),7.52(d,J=2.4Hz,1H),7.33(dd,J=8.8,4.5Hz,1H),7.20(dd,J=9.8,2. 4Hz,1H),7.12(d,J=2.4Hz,1H),6.93(td,J=9.2,2.5Hz,1H),6.80(dd,J=8.7,2.4Hz,1H),6.75(d,J=8.7Hz,1H),4.22–4.17(m,4H).

[0174] 11. Synthesis of Compound S11

[0175]

[0176] The synthesis method refers to compound S3, replacing compound 1d with compound 11a, and the other experimental steps are the same to obtain compound S11. 1 H NMR (400MHz, DMSO) δ10.86(s,1H),9.18(s,1H),8.56(s,1H),8.42(s,1H),7.86(d,J=1.7Hz,1H),7.56– 7.53(m,2H),7.36–7.29(m,2H),7.23(dd,J=9.9,2.4Hz,1H),6.94(td,J=9.2,2.4Hz,1H),4.93(s,2H).

[0177] 12. Synthesis of Compound S12

[0178]

[0179] The synthesis method refers to compound S3, replacing compound 1d with compound 12a, and the other experimental steps are the same to obtain compound S12. 1 H NMR (400MHz, DMSO) δ10.85(s,1H),9.17(s,1H),8.55(s,1H),8.41(s,1H),7.86(d,J=1.3Hz,1H),7.56– 7.54(m,2H),7.36–7.30(m,2H),7.23(dd,J=9.8,2.1Hz,1H),6.94(td,J=9.3,2.2Hz,1H),4.93(s,2H).

[0180] 13. Synthesis of Compound S13

[0181]

[0182] Step 1: The synthesis method refers to compound S3, and compound 13a is used to replace compound 3a. The other experimental steps are the same to obtain compound 13b.

[0183]

[0184] Step 2: Under nitrogen protection, use a sealed tube as a reaction container, mix compound 13b (1 eq) and compound 13c (3 eq) in 1,4-dioxane, add ditriphenylphosphine palladium dichloride (0.05 eq), cuprous iodide (0.1 eq) and N,N-diisopropylethylamine (4 eq), react at 110°C for about 8 hours, wait for the reaction to be complete, cool to room temperature, pour the reaction solution into water, extract with ethyl acetate 3 times, collect the organic phase and purify by column to obtain compound S13. 1 H NMR(400MHz,MeOD)δ7.66(s,1H),7.59(d,J=2.0Hz,1H),7.47(s,1H),7.31(d,J=8.4Hz,1H), 7.20(dd,J=8.5,1.4Hz,1H),7.10(d,J=8.6Hz,1H),7.01(dd,J=8.6,2.1Hz,1H),2.10(s,3H).

[0185] 14. Synthesis of Compound S14

[0186]

[0187] The synthesis method refers to compound S13, replacing compound 13c with compound 14a, and the other experimental steps are the same to obtain compound S14. 1 H NMR (400MHz, DMSO) δ10.98(s,1H),8.72(s,1H),8.61(s,1H),7.71(d,J=2.1Hz,1H),7.64(s,1H),7.51 (d,J=2.3Hz,1H),7.34–7.30(m,2H),7.14–7.09(m,2H),5.27(t,J=5.9Hz,1H),4.30(d,J=5.9Hz,2H).

[0188] 15. Synthesis of Compound S15

[0189]

[0190] Step 1: Under nitrogen protection, compound 15a (1 eq) was dissolved in N,N-dimethylformamide, and ditriphenylphosphine palladium dichloride (0.02 eq), cuprous iodide (0.03 eq), triethylamine (2 eq) and compound 15b (3 eq) were added. The mixture was reacted at room temperature for about two hours. After the reaction was complete, the reaction solution was poured into water, extracted with ethyl acetate three times, and the organic phase was collected and purified by column to obtain compound 15c.

[0191] Step 2: Compound 15c was dissolved in methanol, potassium carbonate (1 eq) was added, and the mixture was reacted at room temperature for about 5 hours. After the reaction was complete, the reaction solution was poured into water, extracted with ethyl acetate for 3 times, and the organic phase was collected and purified by column to obtain compound 15d.

[0192]

[0193] Step 3: Dissolve compound 15d (1 eq) in N,N-dimethylformamide, cool to 0°C and slowly add trifluoroacetic anhydride (1.5 eq). After the addition, move to room temperature to react for about 2 hours. After the reaction is complete, slowly pour the reaction solution into water, filter after the solid is fully precipitated, collect the filter cake, dry it and use it directly in the next step.

[0194] Step 4: The obtained filter cake was suspended in a 4N aqueous sodium hydroxide solution, heated to reflux, and reacted for about 3 hours. After the reaction was complete, it was cooled to room temperature and extracted with diethyl ether three times. The aqueous phase was collected and the pH value was adjusted to 4-5 with 4N aqueous hydrochloric acid. After the solid was fully precipitated, it was filtered, dried, and slurried with an appropriate amount of diethyl ether to obtain compound 15f.

[0195]

[0196] Step 5: The synthesis method refers to compound S3, and compound 15f is used to replace compound 3a. The other experimental steps are the same to obtain compound S15. 1 H NMR (400MHz, DMSO) δ11.01(s,1H),8.72(s,1H),8.63(s,1H),7.71–7.70(m,2H),7.53(d,J=2.3Hz,1H) ,7.32(dd,J=12.0,8.6Hz,2H),7.18(dd,J=8.5,1.4Hz,1H),7.11(dd,J=8.8,2.1Hz,1H),3.94(s,1H).

[0197] 16. Synthesis of Compound S16

[0198]

[0199] Step 1: Compound S15 (1 eq) was dissolved in a mixed solvent of n-butanol and water (volume ratio of 1:1), and then copper sulfate pentahydrate (0.2 eq), sodium ascorbyl palmitate (0.4 eq) and compound 16a (1.2 eq) were added and reacted at room temperature for about 3 hours. After the reaction was complete, the reaction solution was poured into water, extracted with ethyl acetate for 3 times, and the organic phase was collected and purified by column to obtain compound S16. 1 HNMR (400MHz, DMSO) δ8.57(s,1H),7.97(s,1H),7.77(s,1H),7.57-7.56(m,1H),7.41(s,1H),7.37(s,1H),7.30(d,J=8. 8Hz,1H),6.79(s,2H),4.52(t,J=4.8Hz,2H),3.91(t,J=5.0Hz,2H),3.65–3.64(m,2H),3.56–3.54(m,2H),3.37(s,3H).

[0200] 17. Synthesis of Compound S17

[0201]

[0202] The synthesis method refers to compound S16, and compound 17a (refer to J. Med. Chem. 2017, 60, 5407-5423.) is used to replace compound 16a. The other experimental steps are the same to obtain compound S17. 1 H NMR(400MHz,DMSO)δ8.73(s,1H),8.01(s,1H),7.77(s,1H),7.53-7.52(m,1H),7.38(s,1H),7.31–7.29( m,2H),6.75(s,2H),4.48(t,J=6.1Hz,2H),3.68–3.66(m,4H),2.84(t,J=6.1Hz,2H),2.51–2.49(m,4H).

[0203] 18. Synthesis of Compound S18

[0204]

[0205] The synthesis method refers to compound S16, replacing compound 16a with compound 18a, and the other experimental steps are the same to obtain compound S18. 1H NMR (400MHz, DMSO) δ10.84(s,1H),8.81(s,1H),8.68(s,1H),8.39(s,1H),8.13(s,1H),7.73(d,J=1.9Hz,1H),7.55–7.52(m,2H),7.39( d,J=8.5Hz,1H),7.31(d,J=8.7Hz,1H),7.10(dd,J=8.7,2.1Hz,1H),5.09(t,J=5.3Hz,1H),4.44(t,J=5.3Hz,2H),3.84(q,J=5.3Hz,2H).

[0206] 19. Synthesis of Compound S19

[0207]

[0208] The synthesis method refers to compound S13, replacing compound 13c with compound 19a, and the other experimental steps are the same to obtain compound S19. 1 H NMR (400MHz, CD3OD) δ7.57(d,J=1.5Hz,1H),7.55(s,1H),7.43(s,1H),7.25(d,J=8.5Hz,1H),7.12– 7.08(m,2H),7.00(dd,J=8.7,1.9Hz,1H),1.48–1.42(m,1H),0.87–0.83(m,2H),0.72–0.68(m,2H).

[0209] 20. Synthesis of Compound S20

[0210]

[0211] Step 1: Under nitrogen protection, compound 15f (1 eq) was dissolved in N,N-dimethylformamide, and diphenyl (trifluoromethyl) sulfonium trifluoromethanesulfonate (2 eq), cuprous iodide (1 eq), 2,2'-bipyridine (1 eq) and potassium carbonate (1 eq) were added in sequence, and reacted at 60°C for about 5 hours. After the reaction was complete, the mixture was cooled to room temperature, poured into water, extracted with ethyl acetate 3 times, and the organic phase was collected and purified by column to obtain compound 20a.

[0212]

[0213] Step 2: The synthesis method refers to compound S3, and compound 20a is used to replace compound 3a. The other experimental steps are the same to obtain compound S20. 1H NMR (400MHz, CD3OD) δ8.51(s,1H),8.22(s,1H),7.78(d,J=2.0Hz,1H),7.54(d,J=8.5H z,1H),7.43(dd,J=8.5,1.3Hz,1H),7.35(dd,J=8.7,2.1Hz,1H),7.17(d,J=8.7Hz,1H).

[0214] 21. Synthesis of Compound S21

[0215]

[0216] The synthesis method refers to compound S13, replacing compound 13c with compound 21a, and the other experimental steps are the same to obtain compound S19. 1 H NMR (400MHz, CD3OD) δ7.78(s,1H),7.68(s,1H),7.59(d,J=3.3Hz,2H),7.46(s,1H),7.33(d,J=8.4H z,1H),7.23(dd,J=8.5,1.2Hz,1H),7.09(d,J=8.7Hz,1H),7.02(dd,J=8.7,2.1Hz,1H),3.89(s,3H).

[0217] 22. Synthesis of Compound S22

[0218]

[0219] The synthesis method refers to compound S3, replacing compound 3a with compound 22a (see J. Med. Chem. 2019, 62, 11280-11300), and the other experimental steps are the same to obtain compound S20. 1 H NMR (400MHz, CD3OD) δ8.70(s,1H),8.63(s,1H),8.33(s,1H),7.96(d,J=8.2Hz,1H),7.90(d,J=8.7Hz,1 H),7.75(d,J=8.2Hz,1H),7.71(d,J=8.7Hz,1H),4.62–4.55(m,4H),3.93(s,2H),1.82(t,J=6.9Hz,6H).

[0220] 23. Synthesis of Compound S23

[0221]

[0222] Step 1: Under nitrogen protection, compound 13b (1 eq) was dissolved in ultra-dry acetonitrile, and compound 23a (1.5 eq), tri(o-methylphenyl)phosphine (0.2 eq), palladium acetate (0.1 eq) and triethylamine (3 eq) were added in sequence, and reacted at 90°C for about 6 hours. After the reaction was complete, the mixture was cooled to room temperature, poured into water, extracted with ethyl acetate 3 times, and the organic phase was collected and purified by column to obtain compound S23. 1 H NMR (400MHz, DMSO) δ11.07(s,1H),8.82(s,1H),8.61(s,1H),7.81(s,1H),7.74(d,J=15.9Hz,1H),7.71(d,J=1.7Hz,1H),7.53 –7.50(m,2H),7.37(d,J=8.5Hz,1H),7.31(d,J=8.7Hz,1H),7.12(dd,J=8.7,2.2Hz,1H),6.48(d,J=15.9Hz,1H),3.72(s,3H).

[0223] 24. Synthesis of Compound S24

[0224]

[0225] The synthesis method refers to compound S23, replacing compound 23a with compound 24a, and the other experimental steps are the same to obtain compound S24. 1 H NMR (400MHz, DMSO) δ10.84(s,1H),8.78(s,1H),8.55(s,1H),7.71(d,J=1.7Hz,1H),7.52(s,1H),7.47(d,J=2.1Hz,1 H),7.33–7.29(m,3H),7.10(dd,J=8.8,2.1Hz,1H),6.74(d,J=16.0Hz,1H),6.12–6.03(m,1H),3.54(d,J=5.9Hz,2H).

[0226] 25. Synthesis of Compound S25

[0227]

[0228] The synthesis method refers to compound S23, replacing compound 23a with compound 25a, and the other experimental steps are the same to obtain compound S25. 1H NMR (400MHz, DMSO) δ11.13(s,1H),8.86(s,1H),8.59(s,1H),7.71(t,J=8.1Hz,3H),7.53(s,1H),7.49(d, J=8.5Hz,1H),7.39(d,J=8.4Hz,1H),7.31(d,J=8.4Hz,1H),7.11(d,J=8.2Hz,1H),6.23(d,J=16.4Hz,1H).

[0229] 26. Synthesis of Compound S26

[0230]

[0231] Step 1: The synthesis method refers to compound S1, and compound 26a is used to replace compound 1a. The other experimental steps are the same to obtain compound 26d.

[0232]

[0233] Step 2: Dissolve compound 26d in ethanol, add 5% palladium / carbon (mass ratio is 20%), and react at room temperature for about 6 hours under a hydrogen atmosphere. After the reaction is complete, filter out the palladium / carbon, wash the filter cake with an appropriate amount of ethanol, collect the filtrate and spin dry to obtain compound 26e, and use the crude product directly in the next step.

[0234]

[0235] Step 3: Compound 26e (1 eq) was dissolved in dichloromethane, triethylamine (2 eq) and acryloyl chloride (2 eq) were added, and the mixture was reacted overnight at room temperature. After the reaction was complete, the reaction solution was poured into water, extracted with ethyl acetate for 3 times, and the organic phase was collected and purified by column to obtain compound S26. 1 H NMR (400MHz, CD3OD) δ7.98(d,J=1.6Hz,1H),7.57(d,J=1.9Hz,1H),7.42(s,1H),7.32(d,J=8.7Hz,1H),7.21(dd,J=8.7,1.6Hz,1H),7.07 (d,J=8.7Hz,1H),6.99(dd,J=8.7,2.1Hz,1H),6.46(dd,J=17.0,10.0Hz,1H),6.35(dd,J=17.0,1.9Hz,1H),5.74(dd,J=10.0,1.9Hz,1H).

[0236] 27. Synthesis of Compound S27

[0237]

[0238] The synthesis method refers to compound S3, replacing compound 3a with compound 27a, and the other experimental steps are the same to obtain compound S27. 1 H NMR (400MHz, CD3OD) δ7.72(s,1H),7.57(s,1H),7.39–7.28(m,4H).

[0239] Example 2. Inhibitory activity of compounds on STING in human THP1-Dual and mouse Raw-Lucia reporter cells

[0240] 1. The half inhibitory concentration (IC) of the compound on STING activation in human cells 50 )test

[0241] The present invention tests the half inhibitory concentration (IC) of the compounds on STING activation in human THP1-Dual cells. 50 ) to detect the inhibitory activity.

[0242] 1.1 Detection principle: A dual reporter system of interferon-stimulated genes (ISG) and nuclear transcription inflammatory factors (NF-kB) is constructed in the human monocytic cell line THP1-Dual cells. Under the action of activators or inhibitors, the expression of downstream luciferase and alkaline phosphatase can be regulated respectively, and the signal intensity of the corresponding pathway is reflected by adding the corresponding luciferase or alkaline phosphatase detection reagent. In this study, since STING modulators mainly regulate the ISG pathway, we used the ISG pathway reporter system to observe the inhibition of the compound on the ISG pathway stimulated by the STING positive agonist MSA-2 to infer the inhibitory activity of the compound on the STING pathway.

[0243] 1.2 Experimental methods:

[0244] 1) Compounds were prepared into 10 mM stock solutions in DMSO.

[0245] 2) In a 96-well plate, 20 μL of compound diluted with physiological saline was added to each well. According to the initial screening activity of the compound, the starting test concentration (1 μM) and concentration gradient (3-fold dilution) of the compound were set. The stimulation concentration of the STING agonist MSA-2 was 10 μM.

[0246] 3) Count THP1-Dual cells and adjust the cell concentration to 5×10 5 / mL, 180μL of cells were added to each well for incubation. The experiment set up different groups, including: cell blank group (solvent), stimulation group (MSA-2), and drug addition groups with different concentration gradients (MSA-2+compounds). The content of solvent DMSO was 0.2%, and three replicates were set up for each well. 20μL of different groups of reagents were added to each well of cells, so the final volume of each test well was 200μL.

[0247] 4) After incubation in a cell culture incubator at 37°C for 24 hours, 20ul of culture medium was taken from each well and placed in a new 96-well plate with a transparent bottom. Then, the luciferase detection reagent QUANTI-Luciferase detection reagent was added and the Lum fluorescence value was immediately measured using spectra-MAX190 (Molecular Devices).

[0248] 5) Calculate IC 50 . Calculate the inhibition rate first: Inhibition rate (IR, %) = [control (Lum) - drug addition (Lum)] / control (Lum) × 100, then IC 50 The values ​​were calculated by the four-parameter fitting method using GraphPad Prism 8.4.3 software.

[0249] 2. The half inhibitory concentration (IC) of the compound on STING activation in mouse cells 50 )test

[0250] The present invention tests the half inhibitory concentration (IC50) of the compound on STING activation in mouse Raw-Lucia reporter cells. 50 ) to detect the inhibitory activity.

[0251] 2.1 Detection principle: Mouse Raw-Lucia cells are used as experimental subjects. An ISG reporter system is constructed in the cells to regulate the expression of downstream luciferase under the action of activators or inhibitors. The signal intensity of the pathway is then reflected by adding the corresponding luciferase detection reagent. In this study, we observed the inhibition of the ISG pathway stimulated by the STING positive agonist MSA-2 by the compounds to infer the inhibitory activity of the compounds on the STING pathway.

[0252] 2.2 Experimental methods

[0253] 1) Compounds were prepared into 10 mM stock solutions in DMSO.

[0254] 2) In a 96-well plate, 20 ul of compound diluted with physiological saline was added to each well. According to the initial screening activity of the compound, the starting test concentration (1 μM) and concentration gradient (3-fold dilution) of the compound were set. The concentration of the STING agonist MSA-2 was 50 μM.

[0255] 3) Raw-Lucia cell count, adjust the cell concentration to 1×10 6 / ml, 180μl of cells were added to each well for incubation. The experiment set up different groups, including: cell blank group, stimulation group (cell + MSA-2 + solvent), different concentration gradient drug addition groups (cell + MSA-2 + compound), the content of solvent DMSO was 0.6%, and three replicates were set up for each. 20μl of different groups of reagents were added to each well of cells, so the final volume of each test well was 200μl.

[0256] 4) After incubation in a cell culture incubator at 37°C for 24 hours, 20ul of culture medium was taken from each well and placed in a new 96-well plate with a transparent bottom. Then, the luciferase detection reagent QUANTI-Luciferase detection reagent was added and the Lum fluorescence value was immediately measured using spectra-MAX190 (Molecular Devices).

[0257] 5) Calculate IC 50 . Calculate the inhibition rate first: Inhibition rate (IR, %) = [control (Lum) - drug addition (Lum)] / control (Lum) × 100, then IC 50 The values ​​were calculated by the four-parameter fitting method using GraphPad Prism 8.4.3 software.

[0258] The inhibitory activity of the compounds of the present invention on STING in human and mouse cells is shown in Table 2 below.

[0259] Table 2 The half inhibitory concentration (IC) of the compounds of the present invention on STING activation in human and mouse cells 50 )value

[0260]

[0261]

[0262] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in, The compound is a compound represented by Formula Ia or Formula Ib: X is selected from CH, N; n is selected from 1 and 2; R1 is selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, R a Substituted or unsubstituted C1-C8 alkyl, R a Substituted or unsubstituted C2-C8 alkenyl, R a Substituted or unsubstituted C2-C8 alkynyl, R a substituted or unsubstituted aminoacyl; R2 is selected from hydrogen; m is selected from 1, 2, 3, 4; R3 are the same or different and are independently selected from halogen; R a Selected from hydrogen, halogen, hydroxyl, cyano, unsubstituted C2-C8 alkenyl, unsubstituted C1-C8 haloalkyl, unsubstituted C1-C8 alkoxyacyl, unsubstituted 3-6 membered cycloalkyl, substituted or unsubstituted 5-6 membered heteroaryl, wherein the substitution is selected from C1-C8 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that R1 is selected from hydrogen, halogen, R a Substituted or unsubstituted C1-C6 alkyl, R a Substituted or unsubstituted C2-C6 alkenyl, R a Substituted or unsubstituted C2-C6 alkynyl, wherein R a As claimed in claim 1.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: Same as R3.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R a Selected from hydrogen, halogen, hydroxy, cyano, unsubstituted C1-C6 haloalkyl, unsubstituted C1-C6 alkoxyacyl, unsubstituted 3-6 membered cycloalkyl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from the compounds listed in Table 1: Table 1 6. A method for preparing a compound of formula I or a pharmaceutically acceptable salt thereof, characterized in that: The method is selected from the group consisting of: Method 1, comprising the steps of: a-1) reacting the compound of formula 1 in the presence of an azidating agent to obtain a compound of formula 2; a-2) the compound of formula 2 is subjected to rearrangement reaction to obtain a compound of formula 3; a-3) reacting a compound of formula 3 with a compound of formula 4 to obtain a compound of formula I; Wherein, A, R1, R2, R3, m, n, and X are as defined in claim 1; Method 2, including the steps: b) reacting the compound of formula 1 with the compound of formula 4 in the presence of microwaves and an azo reagent to obtain a compound of formula I; wherein A, R1, R2, R3, m, n, and X are as defined in claim 1.

7. A pharmaceutical composition, characterized in that Include: (i) one or more therapeutically effective amounts of the compounds of claim 1 or pharmaceutically acceptable salts thereof; and (ii) a pharmaceutically acceptable carrier.

8. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7, characterized in that: Used for preparing a preparation for preventing and / or treating a disease associated with interferon gene stimulatory protein, wherein the disease associated with interferon gene stimulatory protein is selected from the following group: Singleton-Merten syndrome (SMS), Aicardi-Goutières syndrome (AGS), systemic lupus erythematosus (SLE), familial pernio lupus erythematosus (FCL), retinal vasculopathy and leukodystrophy (RVCL), STING-associated vasculopathy of infancy (SAVI), psoriasis, scleroderma, stroke, brain trauma, cardiovascular disease, diabetes and its complications, neurodegenerative diseases, small intestinal malabsorption syndrome, irritable bowel syndrome, Sjogren's syndrome, multiple sclerosis, non-alcoholic steatohepatitis, rheumatoid arthritis, inflammatory bowel disease, autoimmune colitis, suppurative hidradenitis, uveitis, and mucositis.

9. The use according to claim 8, characterized in that: The cardiovascular disease is selected from the group consisting of atherosclerosis-related vascular disease, myocardial infarction; The neurodegenerative disease is selected from the group consisting of Parkinson's disease, Huntington's disease, and familial amyotrophic lateral sclerosis; The inflammatory bowel disease is selected from the group consisting of Crohn's disease and ulcerative colitis.

10. An inhibitor of interferon gene stimulating protein, characterized in that The inhibitor comprises one or more compounds according to claim 1 or pharmaceutically acceptable salts thereof.

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