4-amino-1-butanone compound as STING agonist

Abstract

The invention discloses a 4-amino-1-butanone compound as an STING agonist, and belongs to the field of medicines. The invention also relates to pharmaceutically acceptable salts, esters, hydrates, solvates, stereoisomers, tautomers, cis-trans-isomers, isotope labels or prodrugs. And compositions comprising such compounds, methods of synthesis of such compounds and use of such compounds in inducing immune responses, inducing STING-dependent type I interferon production and / or treating cell proliferation for preventing and / or treating tumor, immune and / or infectious diseases, such as cancer. # imgabs0 #

Description

Technical Field

[0001] The present disclosure relates to compounds of general formula I or II that can be used as STING (stimulator of interferon genes) agonists for activating the STING pathway, and pharmaceutically acceptable salts, esters, hydrates, solvates, stereoisomers, tautomers, cis-trans isomers, isotope labels or prodrugs thereof. The present disclosure also relates to compositions comprising such compounds, methods for synthesizing such compounds, and uses of such compounds in inducing immune responses, inducing STING-dependent type I interferon production and / or treating cell proliferation, preventing and / or treating tumors, immune and / or infectious diseases, such as cancer. Background Art

[0002] The immune system has evolved to recognize and neutralize different types of threats to maintain host homeostasis and is generally divided into: adaptive and innate. The adaptive immune system specifically recognizes antigens that are not naturally expressed in the host as foreign and generates anti-antigen responses through the coordinated action of leukocyte subsets. A hallmark of adaptive immune responses is their ability to provide "memory" or long-lasting immunity to encountered antigens. While this specific and long-lasting effect is critical to the health and survival of the host, adaptive immune responses take time to develop a full response.

[0003] The innate immune system compensates for this time delay. Innate immunity is the first line of defense against invading pathogens and tissue damage. However, it also responds strongly to certain danger signals associated with cell or tissue damage. The innate immune system has no antigen specificity, but responds to a variety of effector mechanisms. Opsonization, phagocytosis, activation of the complement system, and the production of soluble bioactive molecules (such as cytokines or chemokines) are all mechanisms by which the innate immune system mediates its response. Pattern recognition receptors (PRRs) recognize pathogen-associated molecular patterns (PAMPs) from pathogens or damage-associated molecular patterns (DAMPs) from the body itself, initiating a cascade reaction and inducing the expression of genes such as interferons (IFNs), chemokines, and inflammatory factors. This innate immune response can not only inhibit the proliferation and spread of pathogens in the early stages, but also induce an acquired immune response to ultimately clear the infection. In addition, this innate immune response monitors the occurrence and invasion of tumors. At present, a variety of PRRs have been discovered, such as membrane-bound toll-like receptors (TLRs), C-type lectin receptors (CLRs), non-binding intracellular nucleotide-binding oligomerization domain-like receptors (NLRs), retinoic acid-like receptors (RLRs) and cytoplasmic DNA receptors.

[0004] Pathogen DNA and DNA leaked from the nucleus or mitochondria into the cytoplasm are immune stimulatory molecules, and there are many DNA receptors in the cytoplasm, such as absence in melanoma 2 (AIM2), interferon-inducible protein 16 (IFI16), RNA polymerase III and cyclic GMP-AMP synthase (cGAS). The STimulator of INterferon Genes (STING) located in the endoplasmic reticulum of the cytoplasm is the convergence point of these DNA receptors, controlling the innate immune response mediated by DNA, an immune stimulator. Stimulator of interferon genes (STING) is a key adaptor protein involved in the innate immune response of intracellular nucleic acid ligands, and plays a pivotal role in the T cell-mediated tumor immunity process. Intracellular cyclic guanosine monophosphate-adenosine synthetase (cGAS) can be activated by abnormal double-stranded DNA (dsDNA), catalyzing the conversion of ATP and GTP into atypical 2',3'-cGAMP, activating the STING protein on the endoplasmic reticulum membrane, thereby activating the cGAS-STING pathway and triggering the body's immune response to tumor cells.

[0005] On the one hand, STING agonists can promote the production of type I interferon (IFN), promote cross-presentation and anti-tumor cytotoxic T lymphocyte responses; on the other hand, they can induce the production of chemokines, help recruit effector T cells, and strengthen their immune effects in the tumor microenvironment. In addition, the immune system activated by STING can also increase the body's sensitivity to radiotherapy and achieve the transformation of "cold" tumors to "hot" tumors. Therefore, STING agonists can enhance the immune effect, activate and enhance the body's innate immune system and adaptive immune function from a broader level, and are expected to be suitable for different populations with various types of cancer.

[0006] So far, the STING agonists that have entered the clinical research stage are mainly cyclic dinucleotide (CDN) compounds, such as MK-1454 developed by Merck, ADU-S100 developed by Aduro, and E-7766 developed by Eisai, most of which need to be administered by intratumoral injection. This type of drug has many disadvantages such as large molecular weight, difficulty in synthesis, and poor metabolic stability, which greatly limits the application potential of this type of drug. Most non-CDN STING agonists are in the preclinical research stage, and their activity and drug properties have not yet reached the ideal level.

[0007] Excessive activation of STING can cause the continuous production of cytokines, leading to uncontrollable inflammation and cytokine storm, tissue toxicity, autoimmunity, and an inflammatory tumor microenvironment that promotes tumor growth. Since STING is widely expressed in immune cells and non-immune cells, STING agonists can also induce normal tissues to produce inflammatory cytokines when they are systemically administered. Therefore, the STING agonists currently undergoing clinical trials are administered by intratumoral injection, which limits their application. It is necessary to develop a stable STING agonist that can be administered systemically and has targeting, and it is necessary to accurately control the dosage of the STING agonist to minimize immunotoxicity while exerting its efficacy. Therefore, there is still a need in the art to develop compounds with new structures, good biological activity and high drugability, as well as methods for preventing or treating diseases and conditions such as cancer. The compounds disclosed in the present invention meet this need. Summary of the invention

[0008] The present disclosure relates to novel compounds used as STING agonists and for treating cell proliferative diseases. The present disclosure includes compounds of general I or II and pharmaceutically acceptable salts, esters, hydrates, solvates, stereoisomers, tautomers, cis-trans isomers, isotope labels or prodrugs thereof, which can be used as agents for inducing immune responses, inducing STING-dependent type I interferon production and / or treating cell proliferative diseases.

[0009] A 4-amino-1-butanone-related compound of general formula (I) as a STING agonist, or a pharmaceutically acceptable salt, ester, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof;

[0010] A 4-amino-1-butanone-related compound and a prodrug thereof as a STING agonist, characterized in that the 4-amino-1-butanone-related compound of general formula (I) can undergo a cyclization reaction in vivo to obtain a 3,4-dihydro-2H-pyrrole-related compound of general formula (II) with different biological activities;

[0011]

[0012] W is selected from S and Se;

[0013] Y is selected from OH, OMe or from the following general formula: (IIIa), (IIIb), (IIIc), (IIId) or (IIIe):

[0014]

[0015] wherein Z is selected from O, S or Se;

[0016] Wherein, L is selected from -X1-(CH2)n1-X2-(CH2)n2-X3-: wherein X1, X2, X3 are independently selected from CR3R4, O, or NR5; n1, n2 are each independently 0, 1, 2, 3, 4 or 5;

[0017] Each A-R1 is independently selected from C-R1 and N;

[0018] Each R1 is independently selected from: H, halogen, OR5, N(R5)2, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl substituted by OR5, C1-C6 alkyl substituted by N(R5)2, COOR5 and C(O)N(R5)2;

[0019] Each R2 is independently selected from: H, halogen, CN, OR5, N(R5)2, COOR5, C(O)N(R5)2, SO2R5, C1-

[0020] C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl substituted by OR5, C2-C6 alkenyl, C2-C6 haloalkenyl,

[0021] C2-C6 alkenyl substituted with OR5, C2-C6 alkynyl, C2-C6 haloalkynyl, C2-C6 alkynyl substituted with OR5, C3-C6 cycloalkyl, and 3-6 membered heterocyclic rings comprising 1 to 2 ring members selected from O, S and N(R5);

[0022] Each R3 and R4 is independently selected from the group consisting of: H, halogen, OR5, N(R5)2, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl substituted by OR5, C1-C6 alkyl substituted by N(R5)2, COOR5 and C(O)N(R5)2;

[0023] Each R5 is independently selected from: H, C1-C6 alkyl and C1-C6 haloalkyl;

[0024] In a second aspect, the present invention provides the following compounds, or pharmaceutically acceptable salts, esters, hydrates, solvates, stereoisomers, tautomers, cis-trans isomers, isotope-labeled substances or prodrugs thereof, characterized in that the compounds are selected from but not limited to:

[0025]

[0026]

[0027]

[0028] The compounds of the general formula (I) and (II) of the present invention or their pharmaceutically acceptable salts, wherein the pharmaceutically acceptable salts are inorganic salts or organic salts, the inorganic salts include sodium salts, potassium salts, calcium salts, magnesium salts, iron salts, hydrochlorides, hydrobromides, hydroiodides, sulfates, bisulfates, nitrates, phosphates, and acid phosphates; the organic salts are selected from acetates, trifluoroacetates, propionates, pyruvates, glycolates, oxalates, malonates, fumarates, maleates, lactates, malates, citrates, tartrates, methanesulfonates, sulfonates, benzenesulfonates, and salicylates.

[0029] In a third aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned compound, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, cis-trans isomer, metabolite, isotope-labeled substance, prodrug, or solvate thereof; in the pharmaceutical composition, the excipient includes a pharmaceutically acceptable carrier, diluent, and / or excipient.

[0030] The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into solid preparations for oral administration, including, but not limited to capsules, tablets, pills, powders, granules, etc. In these solid dosage forms, the compounds of the general formula (I) of the present invention are mixed as active ingredients with at least one conventional inert excipient (or carrier), for example, with sodium citrate or dicalcium phosphate. Or mixed with the following ingredients: (1) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (2) binders, such as hydroxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose, gum arabic, etc.; (3) humectants, such as glycerol, etc.; (4) disintegrators, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate, etc.; (5) solubilizers, such as paraffin, etc.; (6) absorption accelerators, such as quaternary ammonium compounds, etc.; (7) wetting agents, such as cetyl alcohol and glyceryl monostearate, etc.; (8) adsorbents, such as kaolin, etc.; (9) lubricants, such as talc, calcium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc., or mixtures thereof. Capsules, tablets and pills may also contain buffers.

[0031] The solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be coated or microencapsulated with coating and shell materials such as enteric coatings and other materials known in the art. They can contain opacifiers, and the release of the active ingredient 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 ingredient can also be formed into microcapsules with one or more of the above-mentioned excipients.

[0032] The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, etc. In addition to the compound of formula (I) or its pharmaceutically acceptable salt as an active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water and other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, in particular cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, etc. or mixtures of these substances, etc. In addition to these inert diluents, the liquid dosage form of the present invention may also include conventional adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and spices, etc.

[0033] The suspending agent includes, for example, ethoxylated stearyl alcohol, polyoxyethylene sorbitol, and sorbitan, microcrystalline cellulose, agar, etc. or a mixture of these substances.

[0034] The compounds of the present invention and their pharmaceutically acceptable salts can be formulated into dosage forms for parenteral injection, including, but not limited to, physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions and dispersions. Suitable carriers, diluents, solvents, excipients include water, ethanol, polyols and suitable mixtures thereof.

[0035] The compound of the present invention or its pharmaceutically acceptable salt can be configured into dosage forms for topical administration, including ointments, powders, suppositories, drops, sprays and inhalants, etc. The compound of the present invention of general formula (I) or its pharmaceutically acceptable salt as an active ingredient is mixed with a physiologically acceptable carrier and optional preservatives, buffers, and propellants that may be required under sterile conditions.

[0036] The pharmaceutical composition of the present invention comprises a compound of the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier, excipient, and diluent. When preparing the pharmaceutical composition, the compound of the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof is usually mixed with a pharmaceutically acceptable carrier, excipient, or diluent. The content of the compound of the general formula (I) or a pharmaceutically acceptable salt thereof can be 0.01-1000 mg, for example 0.05-800 mg, 0.1-500 mg, 0.01-300 mg, 0.01-200 mg, 0.05-150 mg, 0.05-50 mg, etc.

[0037] In a fourth aspect, the present invention provides the above-mentioned compound, or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, cis-trans isomer, metabolite, isotope-labeled substance, prodrug, solvate or the above-mentioned pharmaceutical composition, or the medical use of the above-mentioned pharmaceutical preparation.

[0038] The present invention also provides the use of a compound represented by the general formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable pharmaceutical composition thereof in the preparation of a drug, wherein the drug is used to treat a disease associated with the activity of the STING pathway.

[0039] The present invention also provides the use of the compound represented by general formula (I) or its pharmaceutically acceptable salt or pharmaceutically acceptable pharmaceutical composition in the preparation of drugs for treating autoimmune diseases, infectious diseases, cancers and precancerous syndromes, wherein cancers include melanoma, colon cancer, breast cancer, lung cancer and squamous cell carcinoma.

[0040] Effects of the Invention

[0041] The amino-substituted benzothiophene compound provided by the present invention is a compound represented by formula (I), which has a strong stimulatory effect on the STING-mediated interferon signaling pathway and can be used as a small molecule STING agonist for the clinical treatment of tumors or infectious diseases (especially immunotherapy). DETAILED DESCRIPTION

[0042] The technical solution of the present invention will be described in detail below in conjunction with the embodiments.

[0043] [Definition of terms]

[0044] In the present invention, "C1-C6 alkyl" refers to a saturated linear or branched monovalent hydrocarbon group of 1 to 6 carbon atoms. Examples include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl and 2-methyl-2-propyl.

[0045] In the present invention, "C3-C6 cycloalkyl group" refers to a cycloalkyl group having 3 to 6 carbon atoms.

[0046] In the present invention, "heteroaryl" refers to an unsubstituted or substituted stable 5-membered or 6-membered monocyclic aromatic ring system or an unsubstituted or substituted 9-membered or 10-membered benzo-fused heteroaromatic ring system or a bicyclic heteroaromatic ring system, unless otherwise specified, which consists of carbon atoms and 1-4 heteroatoms selected from N, O or S, and wherein the nitrogen or sulfur heteroatom may be selectively oxidized and the nitrogen heteroatom may be selectively quaternized.

[0047] In the present invention, "substituted" means that one or more hydrogen atoms in the group are replaced by the same or different substituents.

[0048] "Administration" or "administering" a subject compound in the context of the present invention means providing the subject in need of treatment with a compound of the present invention.

[0049] The term "pharmaceutically acceptable salt" refers to salts that are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, etc., and commensurate with a reasonable benefit / risk ratio.

[0050] The term "pharmaceutically acceptable" means that salts, solvents, excipients, etc. are generally non-toxic, safe, and suitable for use by patients. The "patient" is preferably a mammal, more preferably a human.

[0051] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt as defined herein and has all the effects of the parent compound. Pharmaceutically acceptable salts can be prepared by adding the corresponding acid to a suitable organic solvent for an organic base and treating according to conventional methods to prepare pharmaceutically acceptable salts. In the present invention, "pharmaceutically acceptable salts" can be inorganic salts or organic salts, inorganic salts include hydrochlorides, hydrobromides, hydroiodides, sulfates, bisulfates, nitrates, phosphates, acid phosphates, etc., and organic salts include acetates, trifluoroacetates, propionates, pyruvates, glycolates, oxalates, malonates, fumarates, maleates, lactates, malates, citrates, tartrates, methanesulfonates, sulfonates, benzenesulfonates, salicylates, etc.

[0052] The term "hydrate" refers to a substance formed by the combination of a compound of the present invention or a pharmaceutically acceptable salt thereof and water through non-covalent intermolecular forces. Common hydrates include (but are not limited to) hemihydrates, monohydrates, dihydrates, trihydrates, and the like.

[0053] The term "solvate" refers to a substance formed by the combination of a compound of the present invention or a pharmaceutically acceptable salt thereof and at least one solvent molecule through non-covalent intermolecular forces. The term "solvate" includes "hydrates". Common solvates include (but are not limited to) hydrates, ethanolates, acetoneates, etc. It should be understood that the present invention encompasses all solvate forms having PRMT5 inhibitory activity.

[0054] The term "isomers" refers to compounds that have the same number and types of atoms and therefore the same molecular weight, but differ in the arrangement or configuration of the atoms in space.

[0055] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has a vertical asymmetric plane due to at least one chiral factor (including chiral center, chiral axis, chiral plane, etc.), thereby being able to rotate plane polarized light. Since there are asymmetric centers and other chemical structures that may cause stereoisomerism in the compounds of the present invention, the present invention also includes these stereoisomers and mixtures thereof. Since the compounds of the present invention and their salts include asymmetric carbon atoms, they can exist in the form of a single stereoisomer, a racemate, an enantiomer, and a mixture of diastereomers. Generally, these compounds can be prepared in the form of a racemic mixture. However, if desired, such compounds can be prepared or separated to obtain pure stereoisomers, i.e., single enantiomers or diastereomers, or single stereoisomer-enriched (purity ≥98%, ≥95%, ≥93%, ≥90%, ≥88%, ≥85% or ≥80%) mixtures. As described below, a single stereoisomer of a compound is prepared synthetically from an optically active starting material containing the desired chiral center, or by preparing a mixture of enantiomeric products followed by separation or resolution, such as conversion to a mixture of diastereoisomers followed by separation or recrystallization, chromatography, use of a chiral resolution agent, or direct separation of the enantiomers on a chiral chromatographic column. Starting compounds with a specific stereochemistry are either commercially available or prepared as described below and resolved by methods well known in the art. The term "enantiomer" refers to a pair of stereoisomers that are non-superimposable mirror images of each other. The term "diastereomer" or "diastereomers" refers to optical isomers that are not mirror images of each other. The term "racemic mixture" or "racemate" refers to a mixture containing equal parts of a single enantiomer (i.e., an equimolar mixture of two R and S enantiomers). The term "non-racemic mixture" refers to a mixture containing unequal parts of a single enantiomer. Unless otherwise stated, all stereoisomeric forms of the compounds of the present invention are within the scope of the present invention.

[0056] The term "tautomer" (or "tautomeric form") refers to structural isomers with different energies that can be interconverted through a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (or prototropic tautomers) include (but are not limited to) interconversions through proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-imino alcohol isomerization, etc. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

[0057] The term "cis-trans isomer" refers to stereoisomers formed by the different positions of atoms (or groups) located on both sides of a double bond or ring system relative to a reference plane; in cis isomers, the atoms (or groups) are located on the same side of the double bond or ring system, and in trans isomers, the atoms (or groups) are located on opposite sides of the double bond or ring system. Unless otherwise indicated, all cis-trans isomeric forms of the compounds of the present invention are within the scope of the present invention.

[0058] The term "isotope-labeled substance" refers to a compound formed by replacing a specific atom in a structure with an isotope atom thereof. Unless otherwise indicated, the compounds of the present invention include various isotopes of H, C, N, O, F, P, S, and Cl, such as 2H (D), 3H (T), 13C, 14C, 15N, 17O, 18O, 18F, 31P, 32P, 35S, 36S, and 37Cl.

[0059] The term "prodrug" refers to a derivative compound that can directly or indirectly provide a compound of the present invention after being applied to a patient. Particularly preferred derivative compounds or prodrugs are compounds that can increase the bioavailability of the compound of the present invention when administered to a patient (e.g., more easily absorbed into the blood), or compounds that promote the delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise indicated, all prodrug forms of the compounds of the present invention are within the scope of the present invention, and various prodrug forms are well known in the art.

[0060] The term "normal temperature" or "room temperature" means that the temperature may be 10-40°C, or 10-30°C, such as 20°C or 25°C.

[0061] Example

[0062] The embodiments of the present invention will be described in detail below with reference to the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, the process is carried out under conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be purchased from the market.

[0063] Example 1: 4-amino-1-(5,6-dimethoxybenzothiophen-2-yl)butan-1-one (H-1)

[0064]

[0065] Step 1: Dissolve compound 1 (10 g, 47.4 mmol) in DMF (150 mL), cool to 0 ° C, add anhydrous potassium carbonate (13.1 g, 94.8 mmol), then slowly add methyl thioglycolate (7.53 g, 71.1 mmol), and heat to 90 ° C to react overnight. After the reaction is completed, cool to room temperature. Pour the reaction solution into ice water (1 L), solid precipitates, filter, wash the filter cake with water and slurry it with isopropyl ether, and dry the solid to obtain compound 2. (9.5 g, yield: 80%), yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.03(s,1H),7.60(s,1H),7.49(s,1H),3.85(s,6H),3.82(s,3H).

[0066] Step 2: Dissolve compound 2 (9 g, 35.7 mmol) in THF (50 mL) and MeOH (50 mL), add lithium hydroxide (2.57 g, 107.1 mmol) in water (50 mL). Stir overnight at room temperature. After the reaction is completed, concentrate under reduced pressure, dilute the residue with water (100 mL), adjust pH to 2-3 with 1N dilute hydrochloric acid, precipitate solid, filter, wash with water, and dry the solid to obtain compound 3. (8.1 g, yield: 95%), off-white solid. MS-ESI (m / z): 237.2 [M-1] -

[0067] Step 3: Compound 3 (7 g, 29.4 mmol) was dissolved in N-methylpyrrolidone (100 mL), and silver carbonate (12.16 g, 44.1 mmol) and o-phenanthroline hydrate (582 mg, 2.94 mmol) were added, and the temperature was raised to 150 ° C. The reaction was allowed to react for 2 h. After the reaction was completed, the reaction solution was cooled to room temperature and filtered with diatomaceous earth. The filtrate was poured into 500 mL of water, extracted with ethyl acetate 200 mL x 2, and the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 4 (4.2 g, yield: 73%), a white solid. 1 H NMR (400MHz, CDCl3) δ7.31(s,1H),7.27(d,J=5.4Hz,1H),7.24(s,1H),7.20(dd,J=5.4,0.6Hz,1H),3.94(s,3H),3.92(s,3H).

[0068] Step 4: Under nitrogen protection, compound 4 (4 g, 20.6 mmol) was dissolved in ultra-dry tetrahydrofuran (100 mL), cooled to -78 °C, and 1.6 M n-butyl lithium hexane solution (64.3 mL, 103 mmol) was slowly added dropwise. After the addition was completed, the temperature was raised to 0 °C and the reaction was continued for 2 h. Magnesium bromide ether solution (8 g, 30.9 mmol) was slowly added dropwise. After the addition was completed, the reaction solution was raised to room temperature and the reaction was continued for 1 h to obtain a tetrahydrofuran solution of compound 5, which was directly used in the next step without further purification.

[0069] Step 5: Under nitrogen protection, 1-(tert-butyloxycarbonyl)-2-pyrrolidone (7.62g, 41.2mmol) was dissolved in ultra-dry tetrahydrofuran (100mL), cooled to -78°C, and the tetrahydrofuran solution (20.6mmol) of compound 5 prepared in the first step was slowly added dropwise. The reaction solution was reacted at -78°C for 2h and then slowly warmed to room temperature and reacted overnight. After the reaction was completed, 2M hydrochloric acid was slowly added dropwise to the reaction solution for neutralization, and ethyl acetate 200mL x 2 was used for extraction. The organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 6 (3.5g, two-step yield: 45%), a white solid. 1 HNMR (400MHz, CDCl3) δ7.86 (s, 1H), 7.27 (d, J = 4.2Hz, 2H), 4.70 (s, 1H), 4.00 (s, 3H), 3 .97(s,3H),3.29-3.24(m,2H),3.04(t,J=5.4Hz,2H),2.07-1.96(m,2H),1.44(s,9H).

[0070] Step 6: At room temperature, compound 6 (3 g, 7.91 mmol) was dissolved in ethyl acetate (50 mL), and a 4.0 M solution of hydrogen chloride in dioxane (20 mL, 79.1 mmol) was added, and the reaction was allowed to proceed overnight. After the reaction was completed, the reaction solution was concentrated to a small volume at low temperature, a small amount of n-hexane was added, and the crude product was obtained by suction filtration, and compound H-1 was obtained by reverse phase preparative separation as a white solid.

[0071] MS-ESI(m / z):280.1[M+1] +

[0072] 1 H NMR(400MHz,DMSO-d6)δ8.18(s,1H),8.08(s,1H),7.60(s,1H),7.48(s,1H),3.87(s, 3H),3.84(s,3H),3.22-3.18(t,J=5.4Hz,2H),2.92-2.89(m,2H),1.98-1.91(m,2H).

[0073] Example 2: 4-amino-1-(4-fluoro-5,6-dimethoxybenzothiophen-2-yl)butan-1-one (H-2)

[0074]

[0075] Step 1: Under nitrogen protection, compound 7 (10 g, 64.1 mmol) was dissolved in DCM (100 mL), cooled to 0 ° C, and a solution of titanium tetrachloride (21.8 g, 115.4 mmol) in DCM (50 mL) was slowly added dropwise, reacted for 30 min, and a solution of 1,1-dichloromethyl ether (8.84 g, 76.9 mmol) in DCM (20 mL) was slowly added dropwise, reacted for 30 min, warmed to room temperature, and reacted for 5 h. After the reaction was completed, water (100 mL) was added to quench, and dichloromethane 200 mL x 2 was used for extraction. The organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 8 (7.5 g, yield: 63%), a colorless liquid. 1 H NMR (400MHz, CDCl3) δ10.19 (s, 1H), 7.58 (d, J = 9.2Hz, 1H), 6.79 (d, J = 9.2Hz, 1H), 3.95 (s, 6H).

[0076] Step 2: Compound 8 (7 g, 38.0 mmol) was dissolved in toluene (100 mL), rhodanine (5.56 g, 41.8 mmol) and ammonium acetate (5.85 g, 76.0 mmol) were added, the tube was sealed, and the reaction was carried out at 160°C for 20 min. After the reaction was completed, the mixture was cooled to room temperature, and a solid was precipitated. The solid was filtered, washed with water, washed with a small amount of ethanol, and dried to obtain compound 9 (8.1 g, yield: 71%), white solid, MS-ESI (m / z): 300.1 [M+1] +

[0077] Step 3: Compound 9 (8 g, 26.7 mmol) was dissolved in 2 M sodium hydroxide solution (133 mL, 267 mmol), and reacted at 75 °C for 30 min. The reaction solution gradually became clear. After the reaction was completed, the reaction was moved to an ice bath, and 6 M dilute hydrochloric acid was slowly added dropwise to neutralize. Solids precipitated, filtered, and dried to obtain compound 10 (5.3 g, yield: 77%), white solid, MS-ESI (m / z): 259.1 [M+1] +

[0078] Step 4: Compound 10 (5 g, 19.4 mmol) was dissolved in 1,4-dioxane (100 mL), iodine (7.39 g, 29.1 mmol) was added, and the mixture was reacted at 150°C overnight in a sealed tube. After the reaction was completed, the mixture was cooled to room temperature, poured into ice water, extracted with ethyl acetate 100 mL x 2, the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 11 (3.3 g, yield: 66%), white solid, MS-ESI (m / z): 257.1

[0079] [M+1] +

[0080] The synthetic methods of step 5 to step 8 refer to the preparation of compound H-1. Compound H-2 was isolated by reverse phase preparation to obtain a white solid.

[0081] MS-ESI(m / z):298.1[M+1] +

[0082] 1 H NMR(400MHz,DMSO-d6)δ8.00(d,J=5.0Hz,1H),7.11(s,1H),3.90(s,3H),3.85 (s,3H),3.07(t,J=7.6Hz,2H),2.89(tt,J=6.5,5.4Hz,2H),1.93-1.82(m,2H).

[0083] Example 3: 4-amino-1-(5,6-dimethoxybenzo[b]selenophenyl-2-yl)butan-1-one (H-3)

[0084]

[0085] Step 1: Under nitrogen protection, DL-dithiothreitol (6.28 g, 40.8 mmol) and dimethyl diselenide (9.2 g, 49.0 mmol) were dissolved in DMF (150 mL), stirred for 1 h, and then compound 15 (10 g, 40.8 mmol) and DBU (15.5 g, 102 mmol) were added, and the reaction was carried out at room temperature overnight. After the reaction was completed, the reaction solution was poured into ice water (1 L), and the solid precipitated. The solid was filtered, washed with water, and dried to obtain compound 16. (8.2 g, yield: 78%), white solid. 1 H NMR (400MHz, CDCl3) δ10.19(s,1H),7.35(s,1H),7.00(s,1H),3.98(s,3H),3.93(s,3H),2.32(s,3H).

[0086] Step 2: Add compound 16 (8 g, 30.9 mmol) to ethyl bromoacetate (51 g, 309 mmol), and react at 150°C in a sealed tube for 4 h. After the reaction, cool to room temperature, dilute with water (200 mL), extract with ethyl acetate 100 mL x 2, combine the organic phases, wash with water, wash with saturated sodium chloride, dry with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain a crude product, which was purified by column to obtain compound 17 (7.3 g, yield: 71%), a white solid. 1 H NMR (400MHz, CDCl3) δ10.16(s,1H),7.36(s,2H),4.24(q,J=7.1Hz,2H),3.99(s,3H),3.95(s,3H),3.51(s,2H),1.23(t,J=7.2Hz,3H).

[0087] Step 3: Compound 17 (7 g, 21.1 mmol) and anhydrous potassium carbonate (14.4 g, 105.7 mmol) were added to acetonitrile (150 mL), heated to 70 ° C, and reacted for 6 hours. After the reaction, the reaction solution was cooled to room temperature, diluted with water (200 mL), extracted with ethyl acetate 100 mL x 2, and the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 18 (4.1 g, yield: 62%), a white solid. 1 H NMR (400MHz, CDCl3) δ8.17(s,1H),7.31(s,1H),7.26(s,1H),4.40(q,J=7.1Hz,2H),3.99(s,3H),3.95(s,3H),1.40(t,J=7.2Hz,3H).

[0088] The synthetic methods of step 4 to step 8 refer to the preparation of compound H-1. Compound H-3 was isolated by reverse phase preparation to obtain a white solid.

[0089] MS-ESI(m / z):328.1[M+1] +

[0090] 1 H NMR(400MHz,DMSO-d6)δ7.99(s,1H),7.22(s,1H),7.17(s,1H),3.83(s,3H),3.81(s, 3H), 2.87(tt,J=6.6,5.6Hz,2H), 2.81(t,J=7.8Hz,2H), 1.73(tt,J=7.8,5.6Hz,2H).

[0091] Example 4: 4-amino-1-(4-fluoro-5,6-dimethoxybenzo[b]selenophenyl-2-yl)butan-1-one (H-4)

[0092]

[0093] The synthesis of H-4 was based on the preparation of H-3, and a white solid was obtained by reverse phase preparative separation.

[0094] MS-ESI(m / z):346.1[M+1] +

[0095] 1 H NMR(400MHz,DMSO-d6)δ7.84(s,1H),7.38(s,1H),3.89(s,3H),3.86(s,3H),2 .87(tt,J=6.6,5.6Hz,2H),2.81(t,J=7.8Hz,2H),1.73(tt,J=7.8,5.6Hz,2H).

[0096] Example 5: 1,1'-((propane-1,3-diylbis(oxy))bis(6-methoxybenzo[b]thiophene-5,2-diyl))bis(4-aminobutan-1-one) (H-5)

[0097]

[0098] Step 1: Compound 23 (15 g, 46.7 mmol) and ethyl mercaptoacetate (11.2 g, 93.4 mmol) were dissolved in DMF (200 mL), and anhydrous potassium carbonate (12.6 g, 93.4 mmol) and cuprous iodide (4.45 g, 23.3 mmol) were added in sequence, and the temperature was raised to 80°C for reaction for 16 h. After the reaction was completed, water (200 mL) was added for dilution, and ethyl acetate 100 mL x 2 was used for extraction. The organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was slurried with isopropyl ether to obtain compound 24 (12.3 g, yield: 77%), a white solid. 1 H NMR (400MHz, CDCl3) δ7.89 (s, 1H), 7.51-7.28 (m, 7H), 5.23 (s, 2H), 4.42-4.37 (q, J = 5.4Hz, 2H), 4.00 (s, 3H), 1.43-1.40 (t, J = 5.4Hz, 3H).

[0099] The synthetic method of step 2 to step 5 refers to the preparation of compound H-1.

[0100] Step 6: Compound 28 (7.5 g, 16.5 mmol) was dissolved in methanol (100 mL), 10% Pd-C (800 mg) was added, and the mixture was stirred overnight at room temperature under a hydrogen atmosphere (1 atm). After the reaction was completed, the mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was then slurried with ethanol to obtain compound 29 (4.7 g, yield: 78%), a white solid. MS-ESI (m / z): 366.2 [M+1] +

[0101] Step 7: Compound 29 (2 g, 5.48 mmol) and anhydrous potassium carbonate (1.5 g, 11.0 mmol) were added to acetonitrile (50 mL), and then 1,3-dibromopropane (1.1 g, 5.48 mmol) was added and the temperature was raised to 70°C for 6 h. After the reaction, the reaction solution was cooled to room temperature, diluted with water (200 mL), extracted with ethyl acetate 100 mL x 2, and the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 30 (2.1 g, yield: 79%), a white solid. 1 H NMR (400MHz, CDCl3) δ7.85 (s, 1H), 7.31 (s, 1H), 7.28 (s, 1H), 4.24 (t, J = 5.9Hz, 2H), 3.99 (d, J = 7.3Hz, 3H), 3.69 (t, J = 6. 3Hz, 2H), 3.26 (t, J = 6.6Hz, 2H), 3.04 (td, J = 7.2, 2.3Hz, 2H), 2.45 (p, J = 6.1Hz, 2H), 2.00 (p, J = 7.0Hz, 2H), 1.45 (s, 9H).

[0102] Step 8: Compound 30 (500 mg, 1.03 mmol) and anhydrous potassium carbonate (284 mg, 2.06 mmol) were added to acetonitrile (10 mL), and then compound 29 (376 mg, 1.03 mmol) was added and the temperature was raised to 70°C for 6 h. After the reaction, the reaction solution was cooled to room temperature, diluted with water (100 mL), extracted with ethyl acetate 50 mL x 2, and the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 31 (385 mg, yield: 48%), white solid. MS-ESI (m / z): 771.3 [M+1] +

[0103] Step 9: At room temperature, compound 31 (200 mg, 0.26 mmol) was dissolved in ethyl acetate (5 mL), and a 4.0 M solution of hydrogen chloride in dioxane (0.6 mL, 2.6 mmol) was added, and the reaction was allowed to proceed overnight. After the reaction was completed, the reaction solution was concentrated to a small volume at low temperature, and a small amount of n-hexane was added. The solid was precipitated and filtered, and the filter cake was washed with isopropyl ether and dried to obtain a crude product. Compound H-5 was separated by reverse phase preparation to obtain a white solid (76 mg, yield: 51%).

[0104] MS-ESI(m / z):571.2[M+1] +

[0105] 1 H NMR (400MHz, DMSO-d6) δ7.78(d,J=2.1Hz,2H),7.34(d,J=2.1Hz,2H),7.33(s,2H),4.17(t,J=6.4Hz,4H),3.89 (s,6H),3.07(t,J=7.6Hz,4H),2.89(tt,J=6.5,5.4Hz,4H),2.29(p,J=6.5Hz,2H),1.82(tt,J=7.5,5.5Hz,4H).

[0106] Example 6: 4-(5-(3-((2-(4-aminobutyryl)-6-methoxybenzo[b]thiophen-5-yl)oxy)propoxy)-6-methylbenzo[b]thiophen-2-yl)-4-oxobutanoic acid (H-6)

[0107]

[0108] Step 1: Compound 25 (10 g, 31.8 mmol) was dissolved in DMF (150 mL), and dimethylhydroxylamine hydrochloride (3.7 g, 38.2 mmol), EDCI (7.3 g, 38.2 mmol) and triethylamine (22 mL, 159 mmol) were added in sequence, and the mixture was reacted at room temperature for 16 h. After the reaction was completed, water (200 mL) was added for dilution, and ethyl acetate 200 mL x 2 was used for extraction. The organic phases were combined, washed with water, saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 32 (10.4 g, yield: 92%), a white solid. MS-ESI (m / z): 358.1 [M+1] +

[0109] Step 2: Under nitrogen protection, 3-chloro-1-propanol (3.97 g, 42.0 mmol) was dissolved in THF (50 mL), cooled to -20 ° C, and isopropylmagnesium chloride-lithium chloride (1.3M in THF, 55 mL, 42.0 mmol) was slowly added dropwise, reacted for 10 min, magnesium chips (2.02 g, 84.0 mmol) and 1,2-dibromoethane (1 mL) were added, warmed to room temperature, reacted for 20 min, and then warmed to 70 ° C and reacted for 2 h. The reaction was cooled to 0 ° C, and a solution of compound 32 (10 g, 28.0 mmol) in THF (50 mL) was slowly added dropwise, warmed to room temperature, and reacted for 1 h. After the reaction was completed, a saturated ammonium chloride solution was slowly added dropwise to quench the reaction, and ethyl acetate 200 mL x 2 was used for extraction. The organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 33. (6.3 g, yield: 63%), white solid. MS-ESI (m / z): 357.2 [M+1] +

[0110] Step 3: Dissolve chromium trioxide (3.36 g, 33.6 mmol) in water (50 mL), cool to 0°C, and drop concentrated sulfuric acid (8.2 g, 84.0 mmol). Dissolve compound 33 (6 g, 16.8 mmol) in acetone (100 mL), cool to 0°C, and slowly drop the above chromium trioxide aqueous solution. React at 0°C for 1 h, warm to room temperature and react for 2 h. After the reaction is completed, dilute with ice water (200 mL), solid precipitates, filter, wash with water, and dry to obtain compound 34. (4.8 g, yield: 77%), white solid. MS-ESI (m / z): 371.1 [M+1] +

[0111] Step 4: Compound 34 (4.5 g, 12.2 mmol) was dissolved in methanol (50 mL), concentrated sulfuric acid (2.4 g, 24.4 mmol) was added, the temperature was raised to 80°C, and the reaction was continued for 3 h. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated under reduced pressure, diluted with water (100 mL), solid precipitated, filtered, washed with water, and dried to obtain compound 35. (4.1 g, yield: 87%), white solid. MS-ESI (m / z): 385.1 [M+1] +

[0112] The synthetic methods of step 5 to step 7 refer to the preparation of H-5.

[0113] Step 8: Compound 38 (3 g, 4.3 mmol) was dissolved in MeOH (50 mL), and a solution of lithium hydroxide (516 mg, 21.5 mmol) in water (10 mL) was added. Stir at room temperature overnight. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was diluted with water (20 mL), and the pH was adjusted to 2-3 with 1N dilute hydrochloric acid. The solid precipitated, filtered, washed with water, and dried to obtain compound 39. (2.3 g, yield: 78%), off-white solid. MS-ESI (m / z): 686.2 [M+1] +

[0114] The synthesis method of step 9 refers to the preparation of H-1, and H-6 is separated by reverse phase preparation to obtain a white solid. (57 mg, yield: 77%).

[0115] MS-ESI(m / z):586.2[M+1] +

[0116] 1 H NMR (400MHz, DMSO-d6) δ11.52 (s, 1H), 7.78 (dd, J = 2.3, 1.4Hz, 2H), 7.36-7.2 9(m,4H),4.17(t,J=6.4Hz,4H),3.89(s,6H),3.41(t,J=8.5Hz,2H),3.07(t, J=7.6Hz,2H),2.94-2.87(m,2H),2.73(t,J=8.5Hz,2H),2.29(p,J=6.4Hz,2H ),1.82(tt,J=7.5,5.5Hz,2H),1.65(q,J=6.5Hz,1H),1.57(q,J=6.6Hz,1H).

[0117] Example 7: 4-amino-1-(5-(3-((2-(4-aminobutyryl)-6-methoxybenzo[b]selenophen-5-yl)oxy)propoxy)-6-methylbenzo[b]thiophen-2-yl)butan-1-one (H-7)

[0118]

[0119] The synthesis method of H-7 refers to the preparation of H-3 and H-5. A white solid was obtained by reverse phase preparative separation. MS-ESI (m / z): 619.2 [M+1] +

[0120] 1H NMR (400MHz, DMSO-d6) δ7.80(s,1H),7.78(d,J=2.1Hz,1H),7.34(d,J=2.1Hz,1H),7.33(s,1H),7 .22(s,1H),7.14(s,1H),4.19(dt,J=12.7,6.4Hz,4H),3.89(s,3H),3.85(s,3H),3.07(t,J=7.6Hz ,2H),2.88(dtt,J=7.9,6.5,5.6Hz,4H),2.81(t,J=7.8Hz,2H),2.29(p,J=6.4Hz,2H),1.82(tt,J =7.5,5.5Hz,2H),1.73(tt,J=7.8,5.6Hz,2H),1.65(td,J=6.4,3.9Hz,3H),1.57(q,J=6.5Hz,1H).

[0121] Example 8: 4-(5-(3-((2-(4-aminobutyryl)-6-methoxybenzo[b]selenophen-5-yl)oxy)propoxy)-6-methylbenzo[b]thiophen-2-yl)-4-oxobutanoic acid (H-8)

[0122]

[0123] The synthesis method of H-8 refers to the preparation of H-6. White solid. MS-ESI (m / z): 634.1 [M+1] +

[0124] 1 H NMR (400MHz, DMSO-d6) δ11.45(s,1H),7.80(s,1H),7.78(d,J=2.2Hz,1H),7.34(d,J=2.1Hz,1H),7.33(s,1 H),7.22(s,1H),7.14(s,1H),4.19(dt,J=12.6,6.4Hz,4H),3.89(s,3H),3.85(s,3H),3.41(t,J=8.5Hz,2H ), 2.87 (tt, J = 6.6, 5.6 Hz, 2H), 2.81 (t, J = 7.8 Hz, 2H), 2.73 (t, J = 8.5 Hz, 2H), 2.29 (p, J = 6.4 Hz, 2H), 1.73 (tt, J = 7.7, 5.6 Hz, 2H), 1.65 (t, J = 6.4 Hz, 2H). Preparation of tert-butyl (4-(4-fluoro-5-hydroxy-6-methoxybenzothiophen-2-yl)-4-oxobutyl) carbamate (Compound 52)

[0125]

[0126] AlCl3 (514 mg, 5.03 mmol) was added to a mixture of compound 14 (500 mg, 1.26 mmol) in DCM (20 mL) at room temperature. The reaction mixture was stirred for 18 hours. The reaction mixture was then cooled to 0°C and quenched by adding water (50 mL) and 1M HCl. The mixture was extracted with ethyl acetate 50 mL x 2, and the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 52 (312 mg, yield: 64%), white solid, MS-ESI (m / z): 384.1 [M+1] +

[0127] Preparation of tert-butyl (4-(4-chloro-5-hydroxy-6-methoxybenzothiophen-2-yl)-4-oxobutyl) carbamate (Compound 53)

[0128]

[0129] NCS (183 mg, 1.37 mmol) was added to a DMF (5 mL) solution of compound 29 (500 mg, 1.37 mmol) at room temperature. The reaction mixture was then heated to 40 °C and stirred for 2 h. After the reaction was completed, the reaction mixture was cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate 50 mL x 2, the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 53 (341 mg, yield: 62%), white solid, MS-ESI (m / z): 400.1 [M+1] +

[0130] Preparation of (S)-4-(4-fluoro-5-hydroxy-6-methoxybenzo[b]thiophen-2-yl)-2-methyl-4-oxobutanoic acid methyl ester (Compound 56)

[0131]

[0132] Step 1: Dissolve compound 11 (2 g, 7.8 mmol) and oxalyl chloride (1 mL, 11.7 mmol) in tetrahydrofuran (40 mL) under ice bath, add 1 drop of DMF. Stir the reaction mixture at 0°C for 1 h. After the reaction is completed, concentrate the reaction mixture under reduced pressure to obtain compound 54, which is used directly in the next step.

[0133] Step 2: Under nitrogen protection, slowly add (R)-(3-methoxy-2-methyl-3-oxopropyl)zinc (II) bromide solution (0.5M, 31mL, 15.6mmol) to tetrahydrofuran (50mL) containing cuprous chloride (772mg, 7.8mmol) at 0℃. Stir the reaction mixture at 0℃ for 30min. Slowly add compound 54 (2.1g, 7.8mmol) in THF at 0℃, warm to room temperature after addition, and stir for 18 hours. After the reaction is completed, cool the reaction mixture to 0℃ and add saturated NH4Cl aqueous solution (50mL) to quench. Filter, extract with ethyl acetate 100mL x 2, combine the organic phases, wash with water, wash with saturated sodium chloride, dry with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain a crude product, which is purified by column to obtain compound 55 (1.42g, yield: 53%), a white solid.

[0134] The synthesis method of compound 56 refers to the preparation of compound 53. White solid, MS-ESI (m / z): 327.1 [M+1] +

[0135] Preparation of (S)-4,4-difluoro-4-(5-hydroxy-6-methoxybenzothiophen-2-yl)-2-methylbutanoic acid methyl ester (Compound 59)

[0136] Step 1: Under nitrogen protection, compound 57 (1 g, 2.5 mmol) was dissolved in ultra-dry DCM (20 mL), cooled to 0°C, diethylaminosulfur trifluoride (12 g, 75 mmol) was slowly added dropwise, warmed to room temperature, reacted for 1 h, and then heated to 50°C and refluxed for 16 h. After the reaction, the reaction solution was cooled to room temperature, saturated sodium bicarbonate aqueous solution was slowly added dropwise to quench the reaction, and ethyl acetate 50 mL x 2 was extracted, the organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 58 as a white solid, MS-ESI (m / z): 421.1 [M+1] +

[0137] Compound 58 was prepared by hydrogen debenzylation method according to compound 29 to obtain compound 59 as a white solid. MS-ESI (m / z): 331.1 [M+1] +

[0138] Preparation of methyl 4-(5-hydroxy-6-methoxybenzothiophene-2-yl)-2,2-dimethyl-4-oxobutanoate (Compound 62)

[0139]

[0140] The synthesis method of compound 62 refers to the preparation of compound 55. In which (3-methoxy-2,2-dimethyl-3-oxopropyl)zinc (II) bromide is substituted for (R)-(3-methoxy-2-methyl-3-oxopropyl)zinc (II) bromide.

[0141] Preparation of tert-butyl (3-(5-hydroxy-6-methoxybenzo[b]thiophene-2-carboxamide)propyl)carbamate (Compound 64)

[0142] Step 1: Compound 25 (2 g, 6.3 mmol) was dissolved in ultra-dry DMF (20 mL), and tert-butyl (3-aminopropyl)carbamate (1.1 g, 6.3 mmol), HATU (2.8 g, 7.5 mmol) and DMAP (768 mg, 6.3 mmol) were added in sequence. The mixture was reacted at room temperature for 16 hours. After the reaction, water (100 mL) was added for dilution, and ethyl acetate (100 mL x 3) was used for extraction. The organic phases were combined, washed with water, saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. Compound 63 was purified by column chromatography as a white solid. MS-ESI (m / z): 471.2 [M+1] +

[0143] Compound 64 was prepared by hydrogen debenzylation method according to compound 29 to obtain compound 64 as a white solid. MS-ESI (m / z): 381.1 [M+1] +

[0144] Preparation of methyl 4-(5-hydroxy-6-methoxyisoindolin-2-yl)-4-oxobutanoate (Compound 69)

[0145]

[0146] Step 1: Dissolve 4-methoxy-4-oxobutanoic acid (2 g, 15.1 mmol) in DMF (30 mL), add TCFH (6.3 g, 22.6 mmol) and NMI (1.8 g, 22.6 mmol), stir at room temperature for 10 min, then add compound 65 (2.8 g, 15.1 mmol), stir at room temperature for 16 hours, after the reaction is completed, dilute with water (100 mL), extract with ethyl acetate 100 mL x 3, combine the organic phases, wash with water, wash with saturated sodium chloride, dry with anhydrous sodium sulfate, concentrate under reduced pressure to obtain a crude product, and purify by column to obtain compound 66. (2.1 g, yield: 53%), white solid. MS-ESI (m / z): 264.1 [M+1] +

[0147] Step 2: Compound 66 (2 g, 7.6 mmol) was dissolved in acetonitrile (15 mL) and tetrahydrofuran (15 mL), and NBS (2 g, 11.4 mmol) was added, followed by stirring at room temperature for 3 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the crude product was purified by column to obtain compound 67 (1.7 g, yield: 65%), a white solid. MS-ESI (m / z): 342.1 [M+1] +

[0148] Step 3: Compound 67 (1.5 g, 4.4 mmol) was dissolved in dioxane (30 mL), and diboric acid pinacol ester (1.7 g, 6.6 mmol), DPPF palladium dichloride (322 mg, 0.44 mmol) and potassium acetate (647 mg, 6.6 mmol) were added. The reaction solution was reacted at 100 ° C for 16 hours under nitrogen protection. After the reaction, it was concentrated under reduced pressure and the crude product was purified by column to obtain compound 68. (1.1 g, yield: 65%), white solid. MS-ESI (m / z): 390.1 [M+1] +

[0149] Step 4: Sodium perborate (398 mg, 3.9 mmol) was added to tetrahydrofuran (15 mL) and water (15 mL) of compound 68 (1 g, 2.6 mmol), and stirred at room temperature for 1 hour. After the reaction was completed, water (50 mL) was added for dilution, and 50 mL of ethyl acetate was used for extraction. The organic phases were combined, washed with water, saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 69 (0.55 g, yield: 77%), a white solid. MS-ESI (m / z): 280.1 [M+1] +

[0150] Compounds 70 and 71 were prepared following procedures analogous to those outlined for compound 69 using appropriate starting materials either as described in the preparation or obtained from commercial sources.

[0151] Table 1:

[0152]

[0153] Preparation of methyl 4-(4-fluoro-5-hydroxy-6-methoxybenzothiophen-2-yl)-4-oxobutanoate (Compound 73):

[0154]

[0155] The synthesis method of compound 73 refers to the preparation of compound 71. White solid, MS-ESI (m / z): 313.1 [M+1] +Using the corresponding intermediates instead of the synthetic raw materials of Example 5 and Example 6, Examples 9-25 (see Table 1) were synthesized according to the similar operations as Example 5 and Example 6 to obtain the desired products.

[0156] Table 2:

[0157]

[0158]

[0159]

[0160]

[0161] Example 25: (S)-4-(5-(3-(2-(4-aminobutyryl)-6-methoxybenzo[b]thiophen-5-yl)propyl)-6-ethoxyisoindol-2-yl)-2-methyl-4-oxobutanoic acid (H-25)

[0162] Preparation of tert-butyl (4-(5-bromo-6-methoxybenzo[b]thiophen-2-yl)-4-oxobutyl)carbamate (Compound 78)

[0163]

[0164] The synthesis method of compound 78 refers to the preparation of compound 28, white solid, MS-ESI (m / z): 428.1 [M+1] +

[0165] Preparation of (S)-4-(5-(3-(2-(4-aminobutyryl)-6-methoxybenzo[b]thiophen-5-yl)propyl)-6-ethoxyisoindol-2-yl)-2-methyl-4-oxobutanoic acid (H-25)

[0166]

[0167] The methods of step 1 and step 2 refer to the preparation of compound 28.

[0168] Step 3: Compound 80 (2 g, 5.61 mmol) was dissolved in 1,4-dioxane (30 mL), and tetrakistriphenylphosphine palladium (647 mg, 0.56 mmol) and allyl tributyltin (2.8 g, 8.41 mmol) were added. The mixture was protected by nitrogen and heated to 80 °C for 8 hours. After the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by column to obtain compound 81 as a white solid (1.2 g, yield: 67%). MS-ESI (m / z): 318.2 [M+1] +

[0169] Step 4: Compound 81 (1 g, 3.15 mmol) was dissolved in toluene (15 mL), and pinacol borane (806 mg, 6.3 mmol) and LHMDS (0.3 mL, 0.3 mmol, 1 M) were added. The mixture was heated to 100 °C under nitrogen protection and reacted overnight. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by column to obtain compound 82 as a white solid. (0.84 g, yield: 60%). MS-ESI (m / z): 446.2 [M+1] +

[0170] Step 5: Compound 82 (800 mg, 1.8 mmol) and compound 78 (770 mg, 1.8 mmol) were dissolved in 1,4-dioxane (10 mL) and water (1 mL), sodium carbonate (382 mg, 3.6 mmol) was added, nitrogen was protected, and the mixture was heated to 80°C for 4 hours. After the reaction, the mixture was cooled to room temperature, water (20 mL) was added, and ethyl acetate 50 mL x 2 was added for extraction. The organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column to obtain compound 83 (850 mg, yield: 71%), white solid, MS-ESI (m / z): 667.3 [M+1] +

[0171] The synthesis of step 6 and step 7 refers to the preparation of H-4. Compound H-8 was separated by reverse phase preparation to obtain a white solid, MS-ESI (m / z): 553.2 [M+1] +

[0172] 1 H NMR (400MHz, DMSO-d6) δ11.45(s,1H),7.89(d,J=2.2Hz,1H),7.57(d,J=2.1Hz,1H),7.32(s ,1H),7.07(p,J=1.0Hz,1H),6.84(t,J=1.0Hz,1H),4.63-4.49(m,4H),3.82(s,3H),3.81(s, 3H),3.07(t,J=7.6Hz,2H),2.94-2.84(m,3H),2.81-2.64(m,5H),2.59(dd,J=15.2,7.5Hz, 1H),1.89-1.77(m,4H),1.65(q,J=6.5Hz,1H),1.57(q,J=6.6Hz,1H),1.20(d,J=7.8Hz,3H).

[0173] Experimental Example 26: Activation of the STING-mediated interferon (IFN) signaling pathway by the compound

[0174] This experiment evaluates the stimulatory effect of the test compound on the STING-mediated IFN signaling pathway at the cellular level by monitoring the SEAP (secreted embryonic alkaline phosphastase) reporter gene activity regulated by the IFN regulatory factor in THP1-Blue-ISG cells. The test compounds are comparative examples (compounds MSA-1, Refcompound-1) and example compounds (H1-H24).

[0175] The control compound used in this experiment was synthesized according to the prior art method or obtained commercially, and its structural formula is as follows:

[0176]

[0177] In the experiment, THP1-Blue-ISG cells in the logarithmic growth phase were centrifuged, resuspended in cell culture medium to a density of 2×106 cells / mL, and inoculated into a 96-well cell culture plate at 50 μL / well. The mother solution of the test compound was gradiently diluted with cell culture medium to a -2× working concentration dilution (the control compound MSA-2 and the test compound were 10-fold gradient dilutions: 20, 10, 1, 0.1, 0.01 and 0.001 μM), and 50 μL / well of the compound × working concentration dilution was added to the 96-well plate (50 μL of culture medium containing 2% DMSO was added to the negative control well), and the culture plate was placed in a cell culture incubator and incubated for 16 hours. After the cell incubation was completed, 10 μL of the cell culture supernatant was transferred to a 96-well plate, 90 μL / well of QUANTI-Blue solution was added, and the plate was incubated at 37°C for 3 hours, and the absorbance value at 620 nm was read using an ELISA reader. EC 50 The experimental results are shown in Table 3.

[0178] Table 3 Agonistic effect of test compounds on STING-mediated IFN signaling reporter gene

[0179] Compound No. <![CDATA[EC 50 (nM)]]> Compound No. <![CDATA[EC 50 (nM)]]> H-1 2215 H-14 3.4 H-2 2850 H-15 4.2 H-3 3213 H-16 4.0 H-4 2678 H-17 5.1 H-5 4.1 H-18 4.3 H-6 4.6 H-19 4.4 H-7 3.7 H-20 3.8 H-8 4.3 H-21 2.9 H-9 5.0 H-22 3.5 H-10 3.5 H-23 4.0 H-11 4.8 H-24 4.1 H-12 3.6 Comparative Example 1 5108 H-13 3.7 Comparative Example 2 8.2

[0180] Although the present invention has been described by the specific embodiments above, it should not be construed as being limited thereto; rather, the present invention encompasses the general aspects disclosed above. Various modifications and various embodiments are possible without departing from the spirit and scope of the present invention.

Claims

1. A 4-amino-1-butanone compound as a STING agonist or a pharmaceutically acceptable salt, ester, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof; the structure of the compound is shown in the general formula (I):

2. A 4-amino-1-butanone compound as a STING agonist according to claim 1, or a pharmaceutically acceptable salt, ester, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, characterized in that: The 4-amino-1-butanone compound of general formula (I) can undergo a cyclization reaction in vivo to obtain 3,4-dihydro-2H-pyrrole related compounds of general formula (II) with different biological activities; 3. The compound of general formula (I) and general formula (II) or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof as claimed in claim 1 or 2, characterized in that: W is selected from S and Se; Y is selected from OH, OMe or from the following general formula: (IIIa), (IIIb), (IIIc), (IIId) or (IIIe): wherein Z is selected from O, S or Se; Wherein, L is selected from -X1-(CH2)n1-X2-(CH2)n2-X3-: wherein X1, X2, X3 are independently selected from CR3R4, O, or NR5; n1, n2 are each independently 0, 1, 2, 3, 4 or 5; Each A-R1 is independently selected from C-R1 and N; Each R1 is independently selected from: H, halogen, OR5, N(R5)2, C1-C6 alkyl, C1-C6 haloalkyl alkyl, C1-C6 alkyl substituted by OR5, C1-C6 alkyl substituted by N(R5)2, COOR5 and C(O)N(R5)2; Each R2 is independently selected from: H, halogen, CN, OR5, N(R5)2, COOR5, C(O)N(R5)2, SO2R5, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl substituted with OR5, C2-C6 alkenyl, C2-C6 haloalkenyl, C2-C6 alkenyl substituted with OR5, C2-C6 alkynyl, C2-C6 halo substituted alkynyl, C2-C6 alkynyl substituted by OR5, C3-C6 cycloalkyl, and 1 to 2 selected from O, S and A 3- to 6-membered heterocyclic ring as a ring member of N(R5); Each R3, R4 is independently selected from: H, halogen, OR5, N(R5)2, C1-C6 alkyl, C1- C6 haloalkyl, C1-C6 alkyl substituted by OR5, C1-C6 alkyl substituted by N(R5)2, COOR5 and C(O)N(R5)2; Each R5 is independently selected from the group consisting of: H, C1-C6 alkyl and C1-C6 haloalkyl.

4. A compound of formula I or II as claimed in claim 1-3, or a pharmaceutically acceptable salt, ester, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, characterized in that The compound is selected from but not limited to:

5. Use of the compound having the structure represented by general formula (I) and (II) as described in any one of claims 1 to 4, or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, cis-trans isomer, metabolite, isotope-labeled substance, prodrug, solvate or hydrate and an antibody, polypeptide or aptamer, etc., through a cleavable linking bond to prepare a conjugate, and use of the antibody, polypeptide or aptamer conjugate for preventing and / or treating tumors, immune and / or infectious diseases.

6. A pharmaceutically acceptable salt of a compound of formula (I) and (II) according to claims 1-5, wherein the pharmaceutically acceptable salt is an inorganic salt or an organic salt, the inorganic salt comprising sodium salt, potassium salt, calcium salt, magnesium salt, iron salt, hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, acid phosphate; the organic salt is selected from acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, sulfonate, benzenesulfonate, salicylate.

7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, excipient or diluent.

8. Use of the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for preventing and / or treating tumors or infectious diseases.

9. Use of the compound having the structure represented by general formula (I) and (II) as described in any one of claims 1 to 8, or its pharmaceutically acceptable salts, esters, stereoisomers, tautomers, cis-trans isomers, metabolites, isotope-labeled substances, prodrugs, solvates, hydrates, antibody-drug conjugates, polypeptide-drug conjugates, aptamers and other conjugate drugs or pharmaceutical compositions in the preparation of vaccine adjuvants, wherein the vaccine adjuvant is a vaccine adjuvant for preventing and / or treating tumors, immune or infectious diseases.

10. Use of the compound having the structure represented by general formula (I) and (II) according to any one of claims 1 to 9, or its pharmaceutically acceptable salts, esters, stereoisomers, tautomers, cis-trans isomers, metabolites, isotope-labeled substances, prodrugs, solvates, hydrates, antibody-drug conjugates, polypeptide-drug conjugates, aptamers or other conjugate drugs or pharmaceutical compositions in the preparation of interferon gene stimulating protein agonists.

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