MiR-124 inducer as well as preparation method and application thereof

Abstract

The invention relates to a compound as shown in a formula (1) and a preparation method thereof, a composition containing the compound as shown in the formula (1) and / or pharmaceutically acceptable salt of the composition, a preparation method of the composition and / or the pharmaceutically acceptable salt, and application of the composition as a miR-124 inducer in preparation of drugs for treating miR-124-mediated diseases.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and more specifically, to a class of miR-124 inducers with structures as shown in formula (1) and methods for their preparation, as well as the use of such compounds for the treatment, regulation and / or prevention of miR-124-mediated diseases. Background Technology

[0002] MicroRNAs (miRNAs) are a class of endogenous small RNAs, approximately 20-24 nucleotides in length, that play a variety of important regulatory roles within cells. Each miRNA can target multiple genes, and several miRNAs can regulate the same gene, forming a complex regulatory network. This complex regulatory network can regulate the expression of multiple genes through a single miRNA, or it can finely regulate the expression of a specific gene through a combination of several miRNAs. It is estimated that miRNAs regulate one-third of human genes. Although miRNAs account for only 2% of the total number of human genes, they regulate more than 30% of the genes in the human genome, making them a core component of the gene regulatory network [Stem Cells, 2013, 31:2205-2217]. miRNAs participate in regulating processes such as cell proliferation, differentiation, and apoptosis, playing a crucial regulatory role in individual growth and development, as well as the occurrence and development of various diseases, including inflammation and tumors. Increasing research indicates that miRNAs participate in regulating almost all life processes in organisms [Trends MolMed, 2014, 331:1-10].

[0003] MiR-124 is a highly conserved miRNA, expressed in organisms ranging from lower forms like *C. elegans* to higher forms like humans. Current research suggests that the three coding genes for human miR-124 are located at 8p23.1, 8q12.3, and 20q13.33, and all three contain CpG islands in their promoters. Methylation of these islands can silence the miR-124 coding genes [Oncogene, 2012, 31:1609-1622]. MiR-124 is the most abundant miRNA expressed in the central nervous system, and evidence suggests that it is closely related to the repair of damage to the central nervous system. Niina Vuokila et al. found that after brain injury, the hippocampal gene expression network is long-term regulated by miR-124, making it a chronic regulator of gene expression after brain injury [CellMolLife Sci, 2018, 75:4557-4581]. MiR-124 has shown great pharmaceutical potential in Alzheimer's disease. It is specifically highly expressed in the brain, and significantly downregulated in the brains of Alzheimer's patients. MiR-124 can regulate the expression of multiple genes (such as BACE1, Caveolin-1, GSK-3β, 9PTPN1, and DACT1), thereby affecting synaptic plasticity, neuroinflammation, Aβ production, and Tau protein phosphorylation. Furthermore, low expression of miR-124 has been found in tumors such as brain tumors, gastric cancer, breast cancer, cervical cancer, and prostate cancer, indicating its potential for treating various cancers.

[0004] In recent years, miR-124 has attracted much attention due to its anti-inflammatory activity. ABX464 is a small molecule drug whose RNA splicing regulation produces a potent anti-inflammatory and antiviral effect, and it also has the ability to reduce the HIV reservoir. The anti-inflammatory effect of ABX464 is triggered by the molecule binding to the target cap-binding complex, which is located at the 5' end of each intracellular non-coding RNA molecule. This binding leads to the splicing of a long non-coding RNA, thereby inducing the overexpression of the single microRNA product miR-124. miR-124 then initiates a cascade reaction that produces a potent anti-inflammatory effect [Retrovirology, 2015, 12:1-15]. Currently, ABX464 is being developed for the treatment of various inflammatory diseases and viral diseases. These include Crohn's disease, ulcerative colitis, rheumatoid arthritis, HIV / AIDS, and COVID-19 infection.

[0005]

[0006] However, ABX464 still suffers from poor physicochemical properties and poor oral absorption, leaving considerable room for improvement. Therefore, there is an urgent need in this field to develop new miR-124 inducers with high activity, metabolic stability, and / or high safety. Summary of the Invention

[0007] The purpose of this invention is to provide a novel and highly active miR-124 inducer, its preparation method, and its application.

[0008] In a first aspect of the invention, a class of compounds, isomers thereof, polymorphs, pharmaceutically acceptable salts, hydrates or solvates of the general structural formula as shown in formula (1) are provided:

[0009]

[0010] In the formula:

[0011] Indicates a single bond or a double bond;

[0012] n is 0, 1, 2, or 3;

[0013] X 1 X 2 X 3 X 4 and X 5 Each is independently selected from N or CR 4 And X 1 X 2 X 3 X 4 and X 5 It can contain at most 2 N;

[0014] Each R is independently selected from the following group: halogen, CN, nitro, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy groups;

[0015] R 1 and R 2 Each is selected independently from C 1-3 Alkyl and C 1-3 The alkoxy group, wherein the alkyl group and the alkoxy group are each independently substituted by one or more (e.g., 2, 3, 4 or 5) substituents selected from the group consisting of: halogen, hydroxyl, CN, C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy and C 1-3 Halogenated alkoxy groups;

[0016] Or R 1 and R2 The carbon atoms bonded to it together form C 3-6 cycloalkyl or 4-7 membered heterocyclic alkyl;

[0017] R 3 Selected from halogens, hydroxyl groups, CN, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy;

[0018] R 1 R 2 and R 3 Not both C 1-3 alkyl;

[0019] R 4 Selected from H or halogens.

[0020] In another preferred embodiment, each R is independently selected from the group consisting of: F, Cl, Br, Me, Et, OMe, OEt, CF3, CN, OCF3, or NO2.

[0021] In another preferred embodiment, Selected from the following group:

[0022]

[0023] In another preferred embodiment, Selected from the following group:

[0024] In another preferred embodiment, R 3 Selected from the following groups: F, OH, Me, Et, OMe, OEt, CF3, CHF2, CH2CF3, CN, OCF3 or OCH2CF3.

[0025] In various embodiments, representative compounds of the present invention are selected from the following group:

[0026]

[0027]

[0028]

[0029] In a second aspect of the invention, a pharmaceutical composition is provided for treating, modulating, and / or preventing miR-124-mediated related diseases, characterized in that the pharmaceutical composition comprises:

[0030] As an active ingredient, a compound, or an isomer thereof, polymorph thereof, pharmaceutically acceptable salt, hydrate or solvate thereof, as described in the first aspect of the present invention; and

[0031] Optional pharmaceutically acceptable excipients or carriers.

[0032] In a third aspect of the invention, there is provided the use of the compound, or isomers thereof, polymorphs, pharmaceutically acceptable salts, hydrates or solvates thereof, or pharmaceutical compositions of the first aspect of the invention, characterized in that they are used to prepare pharmaceutical compositions for treating, modulating and / or preventing miR-124-mediated related diseases.

[0033] In another preferred embodiment, the miR-124-mediated associated diseases are selected from the group consisting of: Alzheimer's disease, Parkinson's disease, brain tumors, gastric cancer, liver cancer, lung cancer, colorectal cancer, pancreatic cancer, breast cancer, cervical cancer, endometrial cancer, prostate cancer, AIDS, Crohn's disease, ulcerative colitis, rheumatoid arthritis, allergic rhinitis, osteoarthritis, fibrotic diseases, atopic dermatitis, psoriasis, AIDS, and / or COVID-19 infection.

[0034] In a fourth aspect of the invention, a method for treating, modulating, and / or preventing miR-124-mediated related diseases is provided, comprising the steps of administering to an individual in need a compound described in the first aspect of the invention, or an isomer, crystal form, pharmaceutically acceptable salt, hydrate, or solvate thereof, or a pharmaceutical composition described in the second aspect of the invention.

[0035] In another preferred embodiment, the individuals include humans and non-human mammals.

[0036] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0037] Through extensive and in-depth research, and through numerous screenings and tests, the inventors have for the first time discovered a class of compounds of formula (1), which exhibit significant therapeutic effects on miR-124-mediated diseases. The compounds of this invention demonstrate a strong ability to induce miR-124 expression in PMA / IO-stimulated PBMC cells. Based on this, the present invention was completed.

[0038] Compounds of the present invention and their synthesis

[0039] The present invention provides inducers of miR-124, namely compounds of formula (1), or isomers thereof, polymorphs, pharmaceutically acceptable salts (inorganic or organic), hydrates or solvates. Preferably, the compounds of the present invention are as described in the first aspect.

[0040] The present invention also provides a method for preparing compounds of formula (1) of the present invention. The method for preparing compounds of formula (1) of the present invention is described in detail below, but these specific methods do not constitute any limitation on the present invention.

[0041] On the one hand, the compounds described herein are prepared according to methods known in the art. However, the conditions of the method, such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times, are not limited to those explained below. The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in this specification or known in the art, such combinations being readily performed by those skilled in the art. On the other hand, the present invention also provides a method for preparing the compounds represented by the general formula (1):

[0042] Reaction process 1:

[0043]

[0044] Compounds of general formula (1) can be prepared according to reaction process 1, wherein L is a halogen, Z is a halogen or OTf, and R, R 1 R 2 R 3 X 1 X 2 X 3 X 4 X 5 And n is as defined above.

[0045] Reaction process 2:

[0046]

[0047] Compounds of general formula (1) can be prepared according to reaction process 2, wherein R, R 1 R 2 R 3 X 1 X 2 X 3 X 4 X 5 And n is as defined above.

[0048] Reaction process 3:

[0049]

[0050] Compounds of general formula (1) can be prepared according to reaction process 3, wherein R, R 1 R 2 R 3 X 1 X 2 X 3 X 4 X 5 And n is as defined above.

[0051] Related definitions

[0052] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0053] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0054] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, trans-butenedioic acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; as well as salts of amino acids (such as arginine) and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0055] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0056] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 ( 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen atoms to form deuterated compounds. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs generally have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug half-life in vivo. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0057] "Optional" or "optionally" means that the event or condition described below may, but is not required, occur, and the description includes both the scenario in which the event or condition occurs and the scenario in which the event or condition does not occur.

[0058] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents can be arbitrary on a chemically feasible basis.

[0059] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and R has independent options in each case. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0060] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a single bond.

[0061] When one of the variables is selected as a chemical bond, it means that the two groups connected are directly linked. For example, when L in XLY represents a chemical bond, it means that the structure is actually XY.

[0062] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 C 11 and C 12 It also includes any range from n to n+m, such as C 1-12 Including C 1-3 C 1-6 C 1-9 C 3-6 C 3-9 C 3-12 C 6-9 C 6-12 and C 9-12 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-12-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings. They also include any range from n to n+m, such as 3-12-membered rings including 3-6-membered, 3-9-membered, 5-6-membered, 5-7-membered, 6-7-membered, 6-8-membered, and 6-10-membered rings.

[0063] Unless otherwise specified, "C 1-3 "alkyl" is used to denote a straight-chain or branched saturated aliphatic hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Non-limiting examples of alkyl groups include methyl, ethyl, propyl, n-propyl, isopropyl, etc. In this application, "alkyl" is also intended to include substituted alkyl groups, i.e., alkyl groups in which one or more positions are substituted, particularly 1-4 substituents, which may be substituted at any position. "Haloalkyl" refers to an alkyl group as defined herein in which one or more hydrogen atoms are replaced by the same or different halogens. Examples of haloalkyl groups include -CH2Cl, -CH2CF3, -CH2CCl3, perfluoroalkyl groups (e.g., -CF3), etc.

[0064] Unless otherwise specified, "C 3-6 "Cycloalkyl" refers to a saturated cyclic aliphatic hydrocarbon group consisting of 3 to 6 carbon atoms, including monocyclic and bicyclic systems. The C... 3-6 Cycloalkyl groups include C 3-5 C 4-5 and C 5-6Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. C 3-6 Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. A cycloalkyl group may optionally be substituted with one or more substituents, wherein each substituent is independently a hydroxyl, alkyl, alkoxy, halogen, haloalkyl, amino, monoalkylamino, or dialkylamino group.

[0065] Unless otherwise specified, "C 1-3 "Alkoxy" refers to alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule by an oxygen atom. The C 1-3 Alkoxy groups include C 1-2 C2 and C3 alkoxy groups, etc.; C 1-3 Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, etc.

[0066] Unless otherwise specified, "heterocyclic alkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or sulfur, wherein the sulfur atom as a ring atom is optionally oxidized, and the nitrogen atom is optionally quaternized. Preferably, it comprises 3 to 12 ring atoms, wherein 1 to 4 are heteroatoms; more preferably, it comprises 3 to 8 ring atoms, wherein 1 to 3 are heteroatoms; most preferably, it comprises 4 to 6 ring atoms, wherein 1 to 3 are heteroatoms. Unless otherwise specified, the ring system of "heterocyclic alkyl" can be a monocyclic, bicyclic, spirocyclic, or polycyclic ring system. "Heterocyclic alkyl" can be attached to the remainder of the molecule via one or more ring carbons or heteroatoms. Non-limiting examples include azirrobutyl, oxacyclobutyl, thioheterobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiophene (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuranyl-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperidinyl and 2-piperidinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl, hexahydropyridazinyl, homopiperidinyl, homopiperidinyl, etc.

[0067] "Halogen" or "halogenated" refers to fluorine, chlorine, bromine, or iodine.

[0068] “Cyano” refers to -CN.

[0069] Specific pharmaceutical and medical terms

[0070] The term “acceptable,” as used herein, means that a prescription component or active ingredient does not have an excessively harmful effect on health for general therapeutic purposes.

[0071] The terms “treatment,” “treatment process,” or “therapy” as used herein include alleviating, suppressing, or improving symptoms or conditions of a disease; suppressing the development of complications; improving or preventing underlying metabolic syndromes; suppressing the development of a disease or symptom, such as controlling the progression of a disease or condition; reducing a disease or symptom; alleviating a disease or symptom; reducing complications arising from a disease or symptom; or preventing or treating signs arising from a disease or symptom. As used herein, a compound or pharmaceutical composition, when administered, may improve a disease, symptom, or condition, particularly by improving its severity, delaying its onset, slowing its progression, or reducing its duration. Whether administered regularly or intermittently, continuously or intermittently, it may be attributable to or related to the administration.

[0072] "Active ingredient" refers to the compound represented by formula (1), and pharmaceutically acceptable inorganic or organic salts of compounds of formula (1). The compounds of the present invention may contain one or more asymmetric centers (axial chirality) and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers. The asymmetric centers that may exist depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of the present invention. The present invention means including all such isomeric forms of these compounds.

[0073] The terms “compound,” “composition,” “agent,” or “medicine or medicament” may be used interchangeably here, and all refer to a compound or composition that, when applied to an individual (human or animal), can induce a desired pharmaceutical and / or physiological response through local and / or systemic action.

[0074] The term “administered, administering, or administration” here refers to the direct application of the compound or composition described herein, or the application of a prodrug, derivative, or analog of the active compound.

[0075] While the numerical ranges and parameters used to define the broader scope of this invention are approximate values, the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "approximately" may mean that the actual value falls within the acceptable standard error of the mean, as determined by those skilled in the art. Except for experimental examples, or unless explicitly stated otherwise, it is understood that all ranges, quantities, values, and percentages used herein (e.g., to describe material usage, duration, temperature, operating conditions, quantity ratios, and others similar) are modified with "approximately". Therefore, unless otherwise stated, the numerical parameters disclosed in this specification and the accompanying claims are approximate values ​​and are subject to change as needed. At a minimum, these numerical parameters should be understood as the indicated significant digits and values ​​obtained using general rounding.

[0076] Unless otherwise defined in this specification, scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art. Furthermore, unless conflicting with the context, singular nouns used herein include their plural forms, and vice versa.

[0077] route of administration

[0078] The compounds of this invention and their pharmaceutically acceptable salts can be formulated into various preparations, comprising, within a safe and effective range, the compounds of this invention or their pharmaceutically acceptable salts and pharmacologically acceptable excipients or carriers. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective range of the compound is determined based on the age, condition, and duration of treatment of the patient.

[0079] "Pharmaceutically acceptable excipients or carriers" refers to one or more compatible solid or liquid fillers or gelling substances that 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 and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmacologically acceptable excipients or carriers include 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, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0080] When applying the compounds of this invention, they can be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously), or topically.

[0081] 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: (a) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin wax; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in the dosage forms of capsules, tablets, and pills.

[0082] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0083] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0084] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0085] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0086] Compositions for parenteral injection may comprise 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.

[0087] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0088] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0089] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 20–1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

[0090] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0091] The following description will elaborate on the specific aspects, characteristics, and advantages of the aforementioned compounds, methods, and pharmaceutical compositions, making the content of this invention readily apparent. It should be understood that the detailed descriptions and examples described below are specific embodiments and are for reference only. After reading this description, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by this application.

[0092] The structures of the compounds of this invention can be confirmed using conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, that absolute configuration can be confirmed using conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) is used, where the grown single crystals are used to collect diffraction intensity data using a Bruker D8 venture diffractometer with CuKα phase emission as the light source. The scanning method is as follows: After scanning and collecting relevant data, the crystal structure can be further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0093] In all embodiments, 1 H-NMR was recorded using a Varian Mercury 400 NMR spectrometer, and chemical shifts are expressed as δ (ppm). Unless otherwise specified, the silica gel used for separation was 200-300 mesh, and all eluent ratios were by volume.

[0094] The following abbreviations are used in this invention: CHCl3 represents chloroform; Cu represents copper; Cs2CO3 represents cesium carbonate; EA (EtOAc) represents ethyl acetate; DCM represents dichloromethane; Dioxane represents 1,4-dioxane; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; EtOH represents ethanol; Fe represents iron; h represents hours; K2CO3 represents potassium carbonate; LC-MS represents liquid chromatography-mass spectrometry; MeI (CH3I) represents iodomethane; MeOH represents methanol; MS represents mass spectrometry; Na2CO3 represents sodium carbonate; NaH represents sodium hydride; NaOH represents sodium hydroxide; NMR represents nuclear magnetic resonance; Pd / C represents palladium on carbon; Pd(OAc)2 represents palladium acetate; PE represents petroleum ether; POCl3 represents phosphorus oxychloride; PtO2 represents platinum dioxide; rt represents room temperature; SOCl2 represents thionyl chloride; TEA(Et3N) represents triethylamine; TLC represents thin-layer chromatography; THF represents tetrahydrofuran; Tf2O represents trifluoromethanesulfonic anhydride; Tol represents toluene; TsCl represents p-toluenesulfonyl chloride; Xantphos represents 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene; AcOH(HOAc) represents acetic acid; CDI represents carbonyl diimidazole; DIEA(DIEPA) represents diisopropylethylamine; NH4Cl represents ammonium chloride.

[0095] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0096] Preparation Example 1: Synthesis of 6-chloroimidazolo[1,2-a]quinoxaline-4-yltrifluoromethanesulfonate (Intermediate 1)

[0097] Step 1: Synthesis of Intermediate 1-1

[0098] 2,6-Dichloronitrobenzene (2 g, 10.42 mmol) was dissolved in DMF (50 mL), and ethyl imidazole-2-carboxylate (1.46 g, 10.42 mmol) and Cs₂CO₃ (5.08 g, 15.63 mmol) were added. The mixture was stirred at 80 °C and monitored by LC-MS until the reaction was complete. Water (200 mL) was added, and the mixture was extracted with EA (50 mL * 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 30 / 1 to 10 / 1) to give a white solid intermediate 1-1 (2.7 g, yield 88%), ESI-MS m / z: 295.9 [M + H]. + .

[0099] Step 2: Synthesis of intermediates 1-2

[0100] Intermediate 1-1 (2.7 g, 9.15 mmol) was dissolved in HOAc (30 mL), and iron powder (2.56 g, 45.76 mmol) was added. The mixture was stirred at 100 °C, and the reaction was monitored by LC-MS until the starting material was completely reacted. The mixture was filtered, evaporated to dryness to remove HOAc, neutralized with saturated sodium bicarbonate solution, and extracted with EA (50 mL * 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 2 / 1) to give a yellow solid intermediate 1-2 (1.1 g, 55% yield). ESI-MS m / z: 219.8 [M + H] + .

[0101] Step 3: Synthesis of Intermediate 1

[0102] Intermediates 1-2 (1.1 g, 5.02 mmol) and TEA (1.01 g, 10.04 mmol) were dissolved in DCM (30 mL). Under argon protection, Tf₂O (2.12 g, 7.53 mmol) was added with stirring at 0 °C. After the addition was complete, the reaction was stirred at room temperature. LC-MS monitoring showed that the reaction of the starting material was complete. The solution was poured into water (100 mL), extracted with DCM (50 mL * 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 20 / 1) to give a white solid intermediate 1 (1.3 g, yield 74%), ESI-MS m / z: 351.8 [M + H]. + .

[0103] Using ethyl imidazole-2-carboxylate and correspondingly substituted nitrobenzene as raw materials, intermediate 2-10 can be synthesized in a similar manner to intermediate 1.

[0104]

[0105]

[0106] Preparation Example 2: Synthesis of 4,6-dichloro-1,2-dihydroimidazo[1,2-a]quinoxaline (intermediate 11) and 4,9-dichloro-1,2-dihydroimidazo[1,2-a]quinoxaline (intermediate 12)

[0107]

[0108] Step 1: Synthesis of intermediate 11-1

[0109] 3-Chloro-o-phenylenediamine (2 g, 14.03 mmol) was dissolved in diethyl oxalate (20 mL), and the reaction was carried out at 120 °C. The reaction was monitored by LC-MS until the starting material was completely reacted. The solvent was directly evaporated by oil pump, and column chromatography (PE / EA = 10 / 1 to 1 / 1) was performed to give a pale yellow solid intermediate 11-1 (2.2 g, yield 80%), ESI-MS m / z: 196.9 [M+H]. + .

[0110] Step 2: Synthesis of intermediate 11-2

[0111] Intermediate 11-1 (2.2 g, 11.22 mmol) was dissolved in POCl3 (20 mL) and reacted at 100 °C. LC-MS monitoring showed the reaction was complete. The solution was directly concentrated, neutralized with water (50 mL) and saturated sodium bicarbonate solution, and extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 3 / 1) to give a yellow solid intermediate 11-2 (1.6 g, 62% yield). ESI-MS m / z: 232.8 [M+H] + .

[0112] Step 3: Synthesis of intermediate 11-3 and intermediate 11-3-1

[0113] Intermediate 11-2 (1 g, 4.31 mmol) was dissolved in EtOH (20 mL), and ethanolamine (526 mg, 8.62 mmol) was added. The mixture was heated to 70 °C and stirred. LC-MS monitoring showed that the reaction was complete. A large amount of EtOH was directly evaporated, and the solution was added to water (50 mL). Extraction was performed using EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 1 / 1) to give a red solid intermediate 11-3 (400 mg, yield 36%). ESI-MS m / z: 257.9 [M+H] +And red solid intermediate 11-3-1 (520 mg, yield 47%), ESI-MS m / z: 257.9 [M+H] + .

[0114] Step 4: Synthesis of Intermediate 11

[0115] Intermediate 11-3 (400 mg, 1.56 mmol) was dissolved in CHCl3 (20 mL) and SOCl2 (10 mL), and the mixture was stirred at 80 °C. The reaction was monitored by LC-MS until the starting material was completely reacted. The solution was poured into water (50 mL), neutralized with saturated sodium bicarbonate solution, and extracted with CHCl3 (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 5 / 1 to 0 / 1) to give a yellow solid intermediate 11 (280 mg, yield 74%). ESI-MS m / z: 239.9 [M+H] + .

[0116] Step 5: Synthesis of Intermediate 12

[0117] Intermediate 11-3-1 (520 mg, 2.00 mmol) was dissolved in CHCl3 (20 mL) and SOCl2 (10 mL), and the reaction was stirred at 80 °C. The reaction was monitored by LC-MS until the starting material was completely reacted. The solution was poured into water (50 mL), neutralized with saturated sodium bicarbonate solution, and extracted with CHCl3 (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 5 / 1 to 0 / 1) to give a yellow solid intermediate 12 (410 mg, yield 86%). ESI-MS m / z: 239.9 [M+H]. + .

[0118] Using o-phenylenediamines with different substitutions as raw materials, intermediates 13-17 can be synthesized in a similar manner to intermediates 11 and 12.

[0119]

[0120] Preparation Example 3: Synthesis of 6-bromo-1,2-dihydroimidazo[1,2-a]quinoxaline-4-yltrifluoromethanesulfonate (Intermediate 18)

[0121] Using 3-bromo-o-phenylenediamine as a raw material, intermediate 18-1 can be synthesized in a similar manner to intermediate 11.

[0122] Step 1: Synthesis of intermediate 18-2

[0123] Intermediate 18-1 (1 g, 3.51 mmol) was dissolved in THF / H₂O (20 mL / 4 mL), and LiOH (281 mg, 7.02 mmol) was added. The mixture was heated to 60 °C and stirred. LC-MS monitoring showed that the reaction was complete. A large amount of THF was directly evaporated, and the solution was added to water (50 mL). The solution was neutralized to weakly acidic with 1 M HCl, and extracted with EA (30 mL * 3). The organic phases were combined, dried, and concentrated to give a yellow solid intermediate 18-2 (910 mg, 97% yield). ESI-MS m / z: 266.0 [M+H] + .

[0124] Step 2: Synthesis of Intermediate 18

[0125] Intermediate 18-2 (910 mg, 3.42 mmol) and TEA (691 mg, 6.84 mmol) were dissolved in DCM (20 mL). Under argon protection, Tf₂O (1.45 g, 5.13 mmol) was added with stirring at 0 °C. After the addition was complete, the reaction was stirred at room temperature. LC-MS monitoring showed that the reaction of the starting material was complete. The solution was poured into water (100 mL), extracted with DCM (50 mL * 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 3 / 1) to give a white solid intermediate 18 (830 mg, yield 61%). ESI-MS m / z: 397.9 [M + H]. + .

[0126] Intermediate 19 can be synthesized in a similar manner to intermediate 18.

[0127]

[0128] Preparation Example 4: Synthesis of 4-(1-(trifluoromethyl)cyclopropyl)aniline (intermediate 20)

[0129]

[0130] Step 1: Synthesis of intermediate 20-1

[0131] 1-Bromo-4-(1-(trifluoromethyl)cyclopropyl)benzene (2.0 g, 7.55 mmol) and NH₂Boc (3.5 g, 30.18 mmol) were dissolved in Dioxane (50 mL). Pd₂(dba)₃ (690 mg, 0.76 mmol), Xantphos (1.3 g, 2.26 mmol), and cesium carbonate (12.3 g, 37.73 mmol) were added. The mixture was heated to 110 °C and stirred. LC-MS was used to monitor the reaction until the starting material was completely reacted. The solution was poured into water (100 mL), extracted with EA (50 mL x 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 3 / 1) to give a pale yellow solid intermediate 20-1 (2.1 g, 93% yield). ESI-MS m / z: 302.1 [M+H] + .

[0132] Step 2: Synthesis of Intermediate 20

[0133] Intermediate 20-1 (2.1 g, 6.98 mmol) was dissolved in DCM (20 mL), and TFA (5 mL) was added. After the addition was complete, the mixture was stirred at room temperature. LC-MS monitoring showed that the reaction of the starting material was complete. The solution was poured into water (50 mL), and extracted with DCM (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 1 / 1) to give a white solid intermediate 20 (1.2 g, yield 86%). ESI-MS m / z: 202.1 [M+H] + .

[0134] Using bromobenzene with different substitutions as raw materials, intermediates 21-24 can be synthesized in a similar manner to intermediate 20.

[0135]

[0136]

[0137] Preparation Example 5: Synthesis of 4-(1-(difluoromethyl)cyclopropyl)aniline (intermediate 25)

[0138]

[0139] Step 1: Synthesis of Intermediate 25-1

[0140] 1-(4-bromophenyl)cyclopropane-1-onitrile (5 g, 22.50 mmol) was dissolved in DCM (100 mL), and DIBAL-H (1 M, 27 mL, 27.0 mmol) was added dropwise at -78 °C. After the addition was complete, the mixture was slowly brought to room temperature with stirring. The reaction was monitored by LC-MS until the starting material was completely reacted. The reaction was quenched with 1 M hydrochloric acid aqueous solution under ice bath conditions. The mixture was extracted with DCM (50 mL * 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 5 / 1) to give a yellow solid intermediate 25-1 (2.6 g, yield 51%).

[0141] Step 2: Synthesis of intermediate 25-2

[0142] Intermediate 25-1 (1 g, 4.44 mmol) was dissolved in DCM (20 mL), and DAST (716 mg, 4.44 mmol) was added with stirring at room temperature. After the addition was complete, the reaction was stirred at room temperature, and the reaction was monitored by LC-MS until the starting material was completely reacted. The solution was poured into water (50 mL), extracted with DCM (30 mL x 3), the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 20 / 1 to 5 / 1) to give intermediate 25-2 (450 mg, yield 41%) as a yellow solid.

[0143] The subsequent synthesis can be carried out in a similar manner to intermediate 20.

[0144]

[0145]

[0146] Preparation Example 6: Synthesis of 3-fluoro-4-(1-(trifluoromethyl)cyclopropyl)aniline (intermediate 26)

[0147]

[0148] Step 1: Synthesis of intermediate 26-1

[0149] 4-Bromo-2-fluorophenylboronic acid (2 g, 9.14 mmol) and 2-bromo-3,3,3-trifluoropropene (2.40 g, 13.71 mmol) were dissolved in Dioxane / H₂O (30 / 5 mL). Pd(dppf)Cl₂ (666 mg, 0.91 mmol) and potassium phosphate (3.88 g, 18.28 mmol) were added. The mixture was purged with argon and stirred at 90 °C. The reaction was monitored by LC-MS until the reactants were completely reacted. The solution was poured into water (100 mL), extracted with EA (50 mL * 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1) to give a pale yellow solid intermediate 26-1 (1.8 g, 73% yield).

[0150] Step 2: Synthesis of intermediate 26-2

[0151] Intermediate 26-1 (1.8 g, 6.69 mmol) and methyldiphenylsulfonium tetrafluoroborate (2.31 g, 8.03 mmol) were dissolved in THF (30 mL). NaHMDS (1 M, 12 mL, 12.04 mmol) was slowly added in an ice bath, and the mixture was slowly brought to room temperature with stirring. The reaction was monitored by LC-MS until the starting material was completely reacted. The solution was poured into water (100 mL), extracted with EA (50 mL * 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1) to give intermediate 26-2 (1.1 g, 58% yield), which was a pale yellow solid.

[0152] The subsequent synthesis can be carried out in a similar manner to intermediate 20.

[0153]

[0154] Using (6-chloropyridin-3-yl)boronic acid as a raw material, intermediate 27 can be synthesized in a similar manner to intermediate 26.

[0155]

[0156]

[0157] Preparation Example 7: Synthesis of 6-(1-(trifluoromethyl)cyclopropyl)pyridine-3-amine (intermediate 28)

[0158]

[0159] Step 1: Synthesis of intermediate 28-1

[0160] 2,5-Dibromopyridine (5 g, 21.10 mmol) and 4,4,5,5-tetramethyl-2-(3,3,3-trifluoroprop-1-en-2-yl)-1,3,2-dioxoboronoxane (4.41 g, 21.10 mmol) were dissolved in Dioxane / H₂O (50 / 10 mL). Pd(dppf)Cl₂ (1.54 g, 2.11 mmol) and K₂CO₃ (8.75 g, 63.30 mmol) were added, purged with argon, and stirred at 90 °C. The reaction mixture was monitored by LC-MS until the starting material was completely reacted. The solution was poured into water (100 mL), extracted with EA (50 mL x 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 30 / 1) to give a pale yellow solid intermediate 28-1 (3.2 g, 60% yield). ESI-MS m / z: 252.0 [M+H] + .

[0161] It can be synthesized in a similar manner to intermediate 26.

[0162]

[0163] Using the corresponding bromopyrimidine and bromopyrazine as raw materials, intermediates 29-30 can be synthesized in a similar manner to intermediate 28.

[0164]

[0165] Preparation Example 8: Synthesis of 4-(1-(trifluoromethyl)cyclobutyl)aniline (intermediate 31)

[0166]

[0167] Step 1: Synthesis of intermediate 31-1

[0168] 1-(4-bromophenyl)cyclobutanecarboxylic acid (2 g, 7.84 mmol) and 3,3-dimethyl-1-(trifluoromethyl)-1,2-benzoiodoxapentane (3.24 g, 9.80 mmol) were dissolved in EA (250 mL). Ir[dF(CF3)ppy]2(4,4'-dCF3bpy)PF6 (91 mg, 0.08 mmol), CuCN (140 mg, 1.56 mmol), and 4,7-diphenyl-1,10-phenanthroline (781 mg, 2.36 mmol) were added, followed by 2-tert-butyl-1,1,3,3-tetramethylguanidine (670 mg, 3.92 mmol) and water (4.24 g, 235.30 mmol). The mixture was purged with argon, irradiated with a 40 W blue LED, stirred at room temperature, and monitored by LC-MS until the reaction was complete. Pour into water (100 mL), extract with EA (50 mL * 3), combine organic phases, dry, concentrate, and column chromatography (PE / EA = 10 / 1) to give a pale yellow solid intermediate 31-1 (1.18 g, yield 54%).

[0169] The subsequent synthesis can be carried out in a similar manner to intermediate 20.

[0170]

[0171] Using carboxylic acids with different substitutions as raw materials, intermediates 32-33 can be synthesized in a similar manner to intermediate 31.

[0172]

[0173]

[0174] Example 1: Synthesis of 6-chloro-N-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)imidazo[1,2-a]quinoxaline-4-amine (Compound 1)

[0175]

[0176] Intermediate 1 (100 mg, 0.28 mmol), Intermediate 20 (57 mg, 0.28 mmol), Pd(OAc)2 (6 mg, 0.028 mmol), Xantphos (32 mg, 0.056 mmol), and Cs2CO3 (182 mg, 0.56 mmol) were dissolved in Dioxane (5 mL). The reaction was carried out under argon protection at 90 °C. LC-MS monitoring showed that the reaction was complete. The solution was poured into water (20 mL), extracted with EA (20 mL x 3), and the organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 1 / 1) to give a yellow solid compound 1 (65 mg, yield 57%).

[0177] 1 H NMR (400MHz, DMSO-d6): δ10.15(s,1H),8.77(s,1H),8.44(d,J=8.6Hz,2H),8.21(d,J=8.1Hz,1H),7.79(s, 1H),7.68(d,J=7.8Hz,1H),7.50-7.39(m,3H),1.33(t,J=5.6Hz,2H),1.14(s,2H); ESI-MSm / z:403.1[M+H] + .

[0178] Examples 2-33: Synthesis of compounds 2-10, 14-36

[0179] Using intermediates 1-19 and 20-33, as well as other readily available aromatic amines, target compounds 2-10 and 14-36 were obtained through a synthesis method similar to that described in Example 1.

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186] Example 34: Synthesis of 6,8-dichloro-N-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)imidazo[1,2-a]quinoxaline-4-amine (compound 11)

[0187]

[0188] Compound 34 (100 mg, 0.23 mmol) was dissolved in acetone (5 mL), and activated manganese dioxide (199 mg, 2.28 mmol) was added. The mixture was heated to 80 °C. LC-MS monitoring showed that the reaction was complete. The mixture was filtered, concentrated, and subjected to column chromatography (PE / EA = 10 / 1 to 1 / 1) to give a yellow solid, compound 11 (72 mg, yield 72%).

[0189] 1 H NMR (400MHz, DMSO-d6): δ10.23 (s, 1H), 8.80 (d, J = 1.2Hz, 1H), 8.46-8.38 (m, 3H), 7.82-7.7 6(m,2H),7.46(d,J=8.6Hz,2H),1.33(q,J=5.1Hz,2H),1.14(s,2H); ESI-MSm / z:437.1[M+H] + .

[0190] Examples 35-36: Synthesis of compounds 12-13

[0191] Using compounds 35 and 36 as raw materials, target compounds 12-13 were obtained by a similar synthesis method as in Example 34.

[0192]

[0193]

[0194] Example 37: Compound induces miR-124 expression in PBMC cells

[0195] This study evaluated the ability of the compounds of this invention to affect the transcriptional activity of miR-124 in vitro. The expression of miR-124 in Human PBMCs (Sailybio) was analyzed using qPCR.

[0196] Add 1 mL of thawed human PBMCs to 19 mL of culture medium (RPMI 1640 + 10% hFBS + 1‰ 2-mercaptoethanol) and culture for 30 minutes in a cell culture incubator. Collect PBMCs and resuspend them (3.0 x 10⁻⁶ cells) in medium containing 10 ng / mL PMA (Invivogen) and 200 ng / mL iomycin (Invivogen). 6 Cells were incubated in 6-well plates (3 mL / well) for 48 hours. The diluted compound was added to the cells to a final concentration of 0.1 μM, and the cells were collected by centrifugation after 72 hours.

[0197] miRNA was extracted from cells and purified according to the instructions of the MiPure Cell / Tissue miRNA kit (Vazyme). Genomic DNA elimination reaction solution was added to the purified sample, followed by reverse transcription on a GeneAmp PCR instrument. The reaction products were then tested by qPCR using a real-time quantitative PCR instrument (QuantStudio™ 7Flex). Data analysis was performed using QuantStudio 7 software with default settings to automatically calculate Ct values ​​and export the data. The relative expression level of miR-124 was calculated using the following formula: ΔCt = Ct(target gene) - Ct(internal reference gene); ΔΔCt = ΔCt(compound group) - ΔCt(control group); relative mRNA expression = 2^(-ΔΔCt); In this test, miR-191 was used as the internal reference gene, and the control group was the DMSO treatment group.

[0198] Table 1: Results of miR-124 expression induced by compound (0.1 μM) in PBMC cells

[0199]

[0200]

[0201] * indicates ≤10

[0202] ** indicates >10, but ≤30

[0203] *** indicates >30

[0204] As can be seen from the data in the table above, compared with the positive control ABX464, the compounds of the present invention have a strong ability to induce miR-124 expression in PBMC cells. Compared with the DMSO group, most compounds induced miR-124 expression by more than 30 times at a concentration of 0.1 μM.

[0205] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound, or an isomer, polymorph, pharmaceutically acceptable salt, hydrate, or solvate thereof, having a structure as shown in formula (1): In the formula: Indicates a single bond or a double bond; n is 0, 1, 2, or 3; X 1 X 2 X 3 X 4 and X 5 Each is independently selected from N or CR 4 And X 1 X 2 X 3 X 4 and X 5 It can contain at most 2 N; Each R is independently selected from the following group: halogen, CN, nitro, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy groups; R 1 and R 2 Each is selected independently from C 1-3 Alkyl and C 1-3 Alkoxy group, wherein the alkyl group and the alkoxy group are each independently substituted by one or more substituents selected from the group consisting of: halogen, hydroxyl, CN, C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy and C 1-3 Halogenated alkoxy groups; Or R 1 and R 2 The carbon atoms bonded to it together form C 3-6 cycloalkyl or 4-7 membered heterocyclic alkyl; R 3 Selected from halogens, hydroxyl groups, CN, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy; R 4 Selected from H or halogens.

2. The compound, or isomers, polymorphs, pharmaceutically acceptable salts, hydrates or solvates thereof, as claimed in claim 1, wherein each R in formula (1) is independently selected from the group consisting of F, Cl, Br, Me, Et, OMe, OEt, CF3, CN, OCF3 or NO2.

3. The compound, or its isomers, polymorphs, pharmaceutically acceptable salts, hydrates, or solvates as claimed in claim 1, wherein in formula (1), Selected from the following group:

4. The compound, or its isomers, polymorphs, pharmaceutically acceptable salts, hydrates, or solvates as claimed in claim 1, wherein in formula (1), Selected from the following group: Where R 3 The definition is as described in claim 1.

5. The compound, or isomer, polymorph, pharmaceutically acceptable salt, hydrate, or solvate of claim 1, wherein in formula (1), R 3 Selected from the following groups: F, OH, Me, Et, OMe, OEt, CF3, CHF2, CH2CF3, CN, OCF3 or OCH2CF3.

6. The compound, or its isomers, polymorphs, pharmaceutically acceptable salts, hydrates, or solvates according to claim 1, wherein, The compounds are selected from the group consisting of:

7. A pharmaceutical composition for treating, modulating, and / or preventing miR-124-mediated related diseases, characterized in that, The pharmaceutical composition comprises: The active ingredient being any one of claims 1-6, or an isomer, polymorph, pharmaceutically acceptable salt, hydrate, or solvate thereof; and Optional pharmaceutically acceptable excipients or carriers.

8. Use of a compound, isomer, polymorph, pharmaceutically acceptable salt, hydrate, or solvate as claimed in any one of claims 1-6, characterized in that, Used to prepare pharmaceutical compositions for treating, modulating, and / or preventing miR-124-mediated related diseases.

9. The use as described in claim 8, characterized in that, The miR-124-mediated associated diseases are selected from the following group: Alzheimer's disease, Parkinson's disease, brain tumors, gastric cancer, liver cancer, lung cancer, colorectal cancer, pancreatic cancer, breast cancer, cervical cancer, endometrial cancer, prostate cancer, AIDS, Crohn's disease, ulcerative colitis, rheumatoid arthritis, allergic rhinitis, osteoarthritis, fibrotic diseases, atopic dermatitis, psoriasis, AIDS and / or COVID-19 infection.

10. A method for treating, modulating, and / or preventing miR-124-mediated related diseases, characterized in that, The steps include: administering to a desired individual the compound, or any isomer thereof, polymorph thereof, pharmaceutically acceptable salt, hydrate or solvate thereof, or the pharmaceutical composition as described in claim 7.

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