Use of 1-(3-aminopropyl) substituted cyclic amine compound

Abstract

The present invention relates to pharmaceutical use of a 1-(3-aminopropyl) substituted cyclic amine compound. Specifically, the present invention relates to use of a 1-(3-aminopropyl) substituted cyclic amine compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, a solvate thereof, an optically pure isomer thereof, a stereoisomer thereof, or a mixture thereof in the preparation of a drug for treating and / or preventing multiple sclerosis and related diseases thereof.

Description

Uses of 1-(3-aminopropyl) substituted cyclic amine compounds Technical Field

[0001] The present invention relates to the field of medicine, and in particular to the use of 1-(3-aminopropyl) substituted cyclic amine compounds in the preparation of drugs for treating and / or preventing multiple sclerosis and related diseases. Background Art

[0002] Multiple sclerosis (MS) is an immune-mediated disease characterized by chronic inflammatory demyelination of the central nervous system, primarily affecting the spinal cord and cerebral white matter, accompanied by inflammatory infiltration, demyelination, and glial cell changes. Depending on the course of the disease, MS can be divided into relapsing-remitting MS (RRMS), primary progressive MS (PPMS), and secondary progressive MS (SPMS). Clinical symptoms of MS often include limb weakness, visual impairment, and dizziness. Symptoms can peak within a few days and, in severe cases, can lead to paralysis or blindness. In the late stages of MS, cognitive impairment and psychiatric disorders can also develop. Currently, there are approximately 2.3 million cases of MS worldwide, with an incidence rate of approximately 50 per 1 million people in my country, with a significant upward trend in recent years. The average survival time of multiple sclerosis patients is 40 years. They suffer from the profound impact of the disease and its treatment. They face a series of problems such as progressive disability and psychosocial adaptation in the long term. This not only brings great pain to patients, but also imposes an extremely heavy economic burden on society.

[0003] The pathogenesis of multiple sclerosis (MS) remains unclear, with contributing factors involving the central nervous system, immune system, nutrition, and environment. Demyelination, a primary pathological hallmark of MS, is likely caused by an immune response mediated by T and B cells. During the development of MS, the peripheral immune system, mediated by T and B cells, becomes activated, releasing a large number of inflammatory factors that attack the blood-brain barrier (BBB) ​​and infiltrate the central nervous system. Alterations in the inflammatory microenvironment of the central nervous system lead to glial proliferation and excessive secretion of inflammatory factors, which damage oligodendrocytes, causing them to undergo necrosis and apoptosis, triggering myelin shedding. Current drugs for the treatment of MS only have a restorative effect. However, over 10 MS-modifying drugs have been marketed internationally, primarily suppressing the peripheral immune response to reduce disease relapses and delay disability. Fingolimod, a sphingosine-1-phosphate receptor agonist, binds to S1P receptors on lymphocytes, preventing its release from lymphoid tissues and reducing its penetration into the central nervous system, where it can cause multiple sclerosis. However, due to the diverse distribution of S1P in the human body, fingolimod is prone to adverse reactions, such as lymphocytopenia, bradycardia, atrioventricular block, and macular edema. Currently, only interferon beta-1a, beta-1b, and teriflunomide are approved in China. Teriflunomide is the primary active metabolite of the antirheumatic drug leflunomide. It inhibits dihydroorotate dehydrogenase, a key mitochondrial enzyme in the de novo pyrimidine synthesis pathway, reducing activated T and B cell proliferation, central nervous system lymphocyte infiltration, and axonal loss, thereby protecting nervous system function. Due to the limited availability of drugs for preventing relapses of multiple sclerosis in the Chinese market and the lack of effective treatments to prevent axonal damage or myelin repair, multiple sclerosis patients face significant treatment challenges.

[0004] It is reported that CCR5 antagonists can be used to inhibit multiple sclerosis. However, the current CCR5 antagonists for the treatment of multiple sclerosis have problems such as long administration cycle and slow onset of effect, which makes it difficult to actually use CCR5 antagonists in the preparation of drugs for the treatment of multiple sclerosis.

[0005] Therefore, there is an urgent need in the art to develop new drugs that can effectively treat and / or prevent multiple sclerosis and related diseases. Summary of the Invention

[0006] The object of the present invention is to provide use of 1-(3-aminopropyl) substituted cyclic amine compounds in the preparation of drugs for treating and / or preventing multiple sclerosis and related diseases.

[0007] In a first aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, optically pure isomer, stereoisomer, or mixture thereof, for preparing a medicament for treating and / or preventing multiple sclerosis and related diseases;

[0008] The compound of formula (I) has the following structure:

[0009] in,

[0010] W does not exist or is -CH2CH2-; X is N or CR6;

[0011] R1 is selected from a 5-7 membered heteroaryl group which is unsubstituted or substituted by 1-3 substituents, wherein the heteroaryl group contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen, and the substituents are each independently selected from halogen, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched haloalkoxy, -NR 10 R 11 、-C(=O)R 12 , C1-C4 straight or branched chain alkanoyloxy, cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atom to which they are attached form a 5-7 membered ring;

[0012] R 10 and R 11 Each independently selected from H, C1-C4 linear or branched alkyl and -C(=O)R 13 ;

[0013] R 12 Selected from C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched alkoxy, hydroxy, amino (NH2) and C1-C4 straight chain or branched alkylamino;

[0014] R 13 Selected from H and C1-C4 straight or branched chain alkyl;

[0015] R2 is selected from the following groups which are unsubstituted or substituted with 1-3 substituents: C1-C6 straight or branched chain alkyl, C3-C7 cycloalkyl, 4-7 membered heterocyclyl, C6-C12 aryl or 5-7 membered heteroaryl; wherein the substituents are selected from halogen, hydroxyl, C1-C4 straight or branched chain alkyl, C1-C4 straight or branched chain haloalkyl, C1-C4 straight chain or branched chain alkoxy, C1-C4 straight chain or branched chain alkylcarbonyl, C1-C4 straight chain or branched chain haloalkoxy, C1-C4 straight chain or branched chain alkylsulfonyl, C1-C4 straight chain or branched chain alkylsulfonylcarbamoyl, tetrazolyl, cyano, nitro, amino, carboxyl, phenyl and phenoxy;

[0016] R3, R4 and R5 are each independently selected from hydrogen, C1-C6 straight or branched chain alkyl, C1-C6 straight or branched chain haloalkyl and C3-C7 cycloalkyl;

[0017] R6 is selected from hydrogen and C1-C6 straight or branched alkyl;

[0018] Alternatively, R5 and R6 can be connected with Formed together

[0019] R7 is selected from hydrogen, C(=O)R8, C(=O)OR8, C(=O)NR8R9, SO2R8, and the following groups substituted by 1-3 substituents: C1-C6 straight-chain or branched alkyl, C3-C7 cycloalkyl, 4-7 membered heterocyclyl, benzyl, C6-C12 aryl, and 5-7 membered heteroaryl; wherein the substituent is selected from halogen, hydroxy, C1-C4 straight-chain or branched alkoxy, C1-C4 straight-chain or branched alkyl, C1-C4 straight-chain or branched haloalkyl, C1-C4 straight-chain or branched haloalkoxy, cyano, nitro, amino, and carboxyl;

[0020] R8 and R9 are each independently selected from hydrogen, unsubstituted or substituted by 1-3 substituents from the following groups: C1-C6 straight chain or branched alkyl, C3-C7 cycloalkyl, 4-7 membered heterocyclyl, benzyl, C6-C12 aryl and 5-7 membered heteroaryl; wherein the substituents are selected from halogen, hydroxyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched haloalkoxy, cyano, nitro, amino, carboxyl.

[0021] In another preferred embodiment, the compound is selected from the following group:

[0022] In another preferred embodiment, the multiple sclerosis is selected from the group consisting of relapsing-remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, progressive relapsing multiple sclerosis, or a combination thereof.

[0023] In another preferred embodiment, the multiple sclerosis-related disease is a disease selected from the group consisting of neuromyelitis optica, acute disseminated encephalomyelitis, stroke, craniocerebral injury, epilepsy, Alzheimer's disease, Parkinson's disease, immune inflammatory response, and demyelinating injury.

[0024] In another preferred embodiment, multiple sclerosis and related diseases include relapsing-remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, stroke, craniocerebral injury, epilepsy, immune inflammatory response, and demyelinating injury.

[0025] In another preferred embodiment, the compound is used for:

[0026] (1) Repair demyelination in the spinal cord in multiple sclerosis;

[0027] (2) reduce the number of immune cells infiltrating the spinal cord in multiple sclerosis;

[0028] (3) Reduce myeloid deviation in the spinal cord of multiple sclerosis.

[0029] In another preferred embodiment, the pharmaceutically acceptable salt includes a conventional pharmaceutically acceptable salt formed by reacting the compound of the present invention with an inorganic acid or an organic acid. For example, conventional pharmaceutically acceptable salts can be prepared by reacting the compounds of the present invention with inorganic or organic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid and phosphoric acid, and organic acids such as citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid and hydroxy-1,4-ethanesulfonic acid; or sodium salts, potassium salts, calcium salts, aluminum salts or ammonium salts formed by the compounds of the present invention with inorganic bases; or methylamine salts, ethylamine salts or ethanolamine salts formed by the compounds of the present invention with organic bases.

[0030] In another preferred embodiment, the compound is administered by a route selected from the group consisting of oral, rectal, and parenteral.

[0031] In another preferred embodiment, the parenteral administration is selected from the group consisting of intravenous, intramuscular or subcutaneous administration.

[0032] In another preferred embodiment, the oral preparation is selected from the group consisting of capsules, tablets, pills, powders, granules, emulsions, solutions, suspensions, syrups and tinctures.

[0033] The second aspect of the present invention provides a use of a pharmaceutical composition for preparing a medicament for treating and / or preventing multiple sclerosis and related diseases;

[0034] And the pharmaceutical composition comprises:

[0035] (i) a first active ingredient selected from the group consisting of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, optically pure isomer, or stereoisomer thereof;

[0036] (ii) optionally a second active ingredient selected from the group consisting of interferon beta-1a, interferon beta-1b, teriflunomide, fingolimod, copaxone, ofatuzumab, or a combination thereof;

[0037] (iii) a pharmaceutically acceptable carrier;

[0038] The compound of formula (I) is as defined in the first aspect of the present invention.

[0039] In another preferred embodiment, the compound of formula (I) is selected from the following group:

[0040] In another preferred embodiment, in the pharmaceutical composition, the content of the first active ingredient is 0.01-99 wt%, preferably 0.1-90 wt%, based on the total weight of the pharmaceutical composition.

[0041] In another preferred embodiment, the compound of formula (I) is used as a single ingredient in combination with other effective multiple sclerosis treatment methods (surgical treatment, radiotherapy, etc.) in the comprehensive treatment of multiple sclerosis.

[0042] The third aspect of the present invention provides a method for treating and / or preventing multiple sclerosis and related diseases, comprising administering to a subject in need thereof a medically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof;

[0043] The compound of formula (I) is as defined in the first aspect of the present invention.

[0044] In another preferred embodiment, the subject is a primate mammal, such as a human.

[0045] The present invention relates to a drug for treating and / or preventing multiple sclerosis using a 1-(3-aminopropyl)-substituted cyclic amine compound represented by general formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, and mixture thereof as an active ingredient.

[0046] The drug can be introduced into the body by injection, spray, nasal drops, eye drops, permeation, absorption, physical or chemical methods such as intramuscular, intradermal, subcutaneous, intravenous, or mucosal tissues; or can be mixed with or encapsulated in other substances for introduction into the body. If necessary, one or more pharmaceutically acceptable carriers can be added to the drug.

[0047] The carrier includes conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field.

[0048] 1-(3-aminopropyl)-substituted cyclic amine compounds represented by general formula (I) or pharmaceutically acceptable salts, esters, solvates, stereoisomers, tautomers, and prodrugs thereof are used as active ingredients, alone or in combination, or formulated with other drugs, excipients, etc. into various dosage forms, including but not limited to tablets, powders, pills, injections, capsules, films, suppositories, ointments, and granules. The drugs in the above-mentioned various dosage forms can all be prepared according to conventional methods in the pharmaceutical field.

[0049] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form a new technical solution or another preferred embodiment. Each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equal, or similar purpose. Due to space limitations, they will not be detailed here. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 shows the neurological deficit scores and body weight changes of mice in the model group (EAE+DC521022) and the model group (EAE+Vehicle) after immunization; Figure 1A shows that 50 mg / kg of DC521022 and saline were intraperitoneally injected on the day of modeling; Figure 1B shows that 50 mg / kg of DC521022 and saline were intraperitoneally injected after the onset of symptoms; Figure 1C shows that 50 mg / kg of DC521022 and saline were gavage-administered on the day of modeling; Figure 1D shows that 50 mg / kg of DC521022 and saline were gavage-administered after the onset of symptoms.

[0051] FIG2 shows H&E and Luxol fast blue staining of spinal cord tissue sections of mice in the DC521022-treated group and the normal saline-treated group 14 days and 28 days after administration, respectively.

[0052] FIG3 shows the percentages of demyelination and inflammatory cell infiltration in the spinal cords of mice in the DC521022-treated and saline-treated groups.

[0053] Figure 4 shows the results of flow cytometric analysis of myeloid lineage deviation in mouse bone marrow on days 14 and 28 after DC521022 administration. Figure 4A shows the myeloid lineage deviation in mouse bone marrow analyzed by flow cytometer; Figure 4B shows the changes in the proportion and number of hematopoietic stem cells (HSCs) in the femur after 14 days; and Figure 4C shows the changes in the proportion and number of HSCs after 14 days.

[0054] FIG5 shows the clinical score results of the EAE model group administered with the CCR5 antagonist DAPTA. DETAILED DESCRIPTION

[0055] After extensive and intensive research and extensive screening, the inventors unexpectedly discovered for the first time a class of active ingredients that can effectively inhibit multiple sclerosis and related diseases: compounds represented by general formula (I) or pharmaceutically acceptable salts, enantiomers, diastereomers, or racemates thereof. Experiments have shown that the active ingredients of this invention can effectively repair demyelination in the spinal cord of patients with multiple sclerosis, reduce immune cell infiltration and myeloid deviance in the spinal cord, and thus treat and / or prevent multiple sclerosis. This is the basis for the completion of the present invention.

[0056] the term

[0057] "Alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group. For example, "C1-8 alkyl" refers to a straight-chain alkyl group and a branched alkyl group containing 1 to 8 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, etc.

[0058] "Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent. For example, "C3-8 cycloalkyl" refers to a cycloalkyl group comprising 3 to 8 carbon atoms, which is divided into monocyclic cycloalkyl and polycyclic cycloalkyl. Monocyclic cycloalkyl includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Polycyclic cycloalkyl includes spirocyclic, fused ring, and bridged ring cycloalkyl.

[0059] "Heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent in which one or more ring atoms are selected from nitrogen, oxygen or sulfur.

[0060] "Aryl" refers to an all-carbon monocyclic or fused polycyclic (ie, rings which share adjacent pairs of carbon atoms) group, a polycyclic (ie, rings which have adjacent pairs of carbon atoms) group having a conjugated pi electron system, including but not limited to phenyl and naphthyl.

[0061] "Heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms, including nitrogen, oxygen and S. For example, a 5-7 membered heteroaryl refers to a heteroaromatic system containing 5-7 ring atoms, and a 5-10 membered heteroaryl refers to a heteroaromatic system containing 5-10 ring atoms, including but not limited to furanyl, thienyl, pyridinyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc.

[0062] "Alkoxy" refers to -O-(alkyl), where alkyl is as defined above. For example, "C1-8 alkoxy" refers to an alkyloxy group containing 1 to 8 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, butoxy, etc. The present invention will be further described below with reference to specific examples.

[0063] "Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel 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 the compound of general formula (I) of the present invention, its pharmaceutically acceptable salt or solvate thereof, and each other without significantly reducing the efficacy of the active ingredient. Some examples of pharmaceutically acceptable 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, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0064] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0065] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances. In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings and spices.

[0066] In addition to the active ingredients, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0067] There is no particular limitation on the administration of the compound of formula (I) or the pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and the like.

[0068] 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.

[0069] Active ingredient

[0070] The compound of formula (I) of the present invention has the following structure:

[0071] wherein each group is as defined above.

[0072] In another preferred embodiment, the compound is in S configuration or R configuration, preferably, in S configuration.

[0073] In another preferred embodiment, R1 is selected from the following groups which are unsubstituted or substituted with 1-3 substituents: The substituent is selected from halogen, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched alkoxy, -NR 10 R 11 、-C(=O)R 12 , C1-C4 straight or branched alkylcarbonyloxy, C1-C4 straight or branched halogenated alkoxy, cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atom to which they are connected form a 5-7 membered ring; preferably, the substituent is selected from halogen, C1-C2 alkyl, C1-C2 haloalkyl, C1-C2 alkoxy, NR 10 R 11 、-C(=O)R 12 , C1~C2 alkylcarbonyloxy, C1~C2 haloalkoxy, cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atoms to which they are attached constitute a 5-7 membered carbocyclic ring, a 5-7 membered heteroaromatic ring or a 5-7 membered heterocyclic ring; preferably, the substituents are selected from halogen, methyl, methoxy, ethyl, amino, hydroxyl, cyano, nitro, acetyl, formamido, acetamido, carbamoyl, N-methylcarbamoyl, N,N-dimethylcarbamoyl, formyloxy, acetoxy, methoxycarbonyl, trifluoromethyl and trifluoromethoxy, or two adjacent substituents together with the carbon atoms to which they are attached constitute a benzene ring, a cyclopentene ring or a dioxole ring.

[0074] In another preferred embodiment, R 10 and R 11 Each independently selected from H, C1-C2 alkyl and -C(=O)R 13 .

[0075] In another preferred embodiment, R 12 Selected from C1-C2 alkyl, C1-C2 alkoxy, hydroxy, amino (NH2) and C1-C2 alkylamino.

[0076] In another preferred embodiment, R 13 Selected from H and C1-C2 straight or branched chain alkyl.

[0077] In another preferred embodiment, R2 is selected from the following groups which are unsubstituted or substituted by 1-3 substituents: C1~C4 straight chain or branched alkyl, C3~C7 cycloalkyl, 4~7 membered heterocyclic group, and phenyl, wherein the substituent is selected from halogen, hydroxyl, C1~C4 straight chain or branched alkyl, C1~C4 straight chain or branched haloalkyl, C1~C4 straight chain or branched alkoxy, C1~C4 straight chain or branched alkylcarbonyl, C1~C4 straight chain or branched haloalkoxy, C1~C4 straight chain or branched alkylsulfonyl, C1~C4 straight chain or branched alkylsulfonylcarbamoyl, tetrazolyl, cyano, nitro, amino, carboxyl, phenyl, halophenyl, phenoxy and halophenoxy.

[0078] In another preferred embodiment, R2 is selected from C1-C4 straight or branched alkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyran-4-yl, 1-methylpiperidin-4-yl, 1-acetylpiperidin-4-yl, 1-methylsulfonylpiperidin-4-yl, 4-fluorobenzyl, phenyl, difluoro-substituted cyclohexyl (such as 4,4-difluoro-substituted cyclohexyl), ethylcyclohexyl and phenoxymethyl.

[0079] In another preferred embodiment, R3, R4 and R5 are each independently selected from hydrogen, C1-C4 linear or branched alkyl and C3-C7 cycloalkyl.

[0080] In another preferred embodiment, R3, R4 and R5 are each independently selected from hydrogen, methyl, trifluoromethyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0081] In another preferred embodiment, R3, R4 and R5 are each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl and cyclopropyl.

[0082] In another preferred embodiment, R6 is selected from hydrogen and C1-C4 linear or branched alkyl, more preferably selected from hydrogen, methyl and ethyl.

[0083] Or R5 and R6 can be connected with Formed together

[0084] In another preferred embodiment, R7 is selected from hydrogen, C(=O)R8, C(=O)OR8, C(=O)NR8R9, SO2R8 and the following groups substituted by 1-3 substituents: C1~C4 straight chain or branched alkyl, C3~C7 cycloalkyl, 4-7 membered heterocyclic group, benzyl and phenyl, wherein the substituent is selected from halogen, hydroxyl, C1~C4 straight chain or branched alkoxy, C1~C4 straight chain or branched alkyl, C1~C4 straight chain or branched haloalkyl, C1~C4 straight chain or branched haloalkoxy, cyano, nitro, amino and carboxyl.

[0085] In another preferred embodiment, R7 is selected from C(=O)R8, C(=O)OR8 and SO2R8;

[0086] R8 and R9 are each independently selected from hydrogen and the following groups which are unsubstituted or substituted by 1-3 substituents: C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C3-C7 cycloalkyl, 4-7 membered heterocyclyl, benzyl, phenyl and 5-7 membered heteroaryl, wherein the substituents are selected from halogen, hydroxyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched haloalkoxy, cyano, nitro, amino and carboxyl, preferably selected from halogen, hydroxyl, methoxy, ethoxy, methyl, ethyl, trifluoromethyl, trifluoromethoxy, cyano, nitro, amino and carboxyl.

[0087] In another preferred embodiment, R8 and R9 are each independently selected from hydrogen, C1-C4 linear or branched alkyl, C1-C4 linear or branched halogenated alkyl, C3-C7 cycloalkyl, benzyl and phenyl.

[0088] In another preferred embodiment, R8 and R9 are each independently selected from methyl, ethyl, n-propyl, cyclopropyl, isopropyl, n-butyl, sec-butyl and tert-butyl.

[0089] In another preferred embodiment, a 1-(3-aminopropyl)-substituted cyclic amine compound represented by general formula (II), a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or mixture thereof is provided:

[0090] Wherein, R1, R2, R3, R4, R5 and W are defined as described in the general formula (I).

[0091] In another preferred embodiment, in the general formula (II), R1 is selected from the following groups which are unsubstituted or substituted with 1-3 substituents: The substituent is selected from halogen, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched halogenated alkyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched alkylcarbonyloxy, C1-C4 straight chain or branched halogenated alkoxy, NR 10 R 11 、-C(=O)R 12 , cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atom to which they are connected form a 5-7 membered ring; preferably, the substituent is selected from halogen, C1-C2 alkyl, C1-C2 haloalkyl, C1-C2 alkylcarbonyloxy, C1-C2 alkoxy, C1-C2 haloalkoxy, NR 10 R11 、-C(=O)R 12 , cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atoms to which they are attached form a 5-7 membered carbocyclic ring, a 5-7 membered heteroaromatic ring or a 5-7 membered heterocyclic ring; preferably, the substituents are selected from halogen, methyl, trifluoromethyl, trifluoromethoxy, methoxy, ethyl, amino, cyano, nitro, acetyl, formamido, acetamido, carbamoyl, N-methylcarbamoyl, N,N-dimethylcarbamoyl, acetoxy, formyloxy and methoxycarbonyl, or two adjacent substituents together with the carbon atoms to which they are attached form a benzene ring, a cyclopentene ring or a dioxole ring;

[0092] R 10 and R 11 Each independently selected from H, C1-C4 linear or branched alkyl and -C(=O)R 13 Preferably, R 10 and R 11 Each independently selected from H, C1-C2 alkyl and -C(=O)R 13 ;

[0093] R 12 is selected from C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched alkoxy, hydroxy, amino (NH2) and C1-C4 straight chain or branched alkylamino; preferably, R 12 Selected from C1-C2 alkyl, C1-C2 alkoxy, hydroxy, amino (NH2) and C1-C2 alkylamino;

[0094] R 13 is selected from H and C1-C4 straight or branched alkyl; preferably, R 13 Selected from H and C1-C2 straight or branched chain alkyl;

[0095] R2 is selected from phenyl, C1~C4 straight or branched alkyl and C3~C7 cycloalkyl which are unsubstituted or substituted by 1-3 substituents, wherein the substituents are selected from halogen, hydroxyl, C1~C4 straight or branched alkyl, C1~C4 straight or branched haloalkyl, C1~C4 straight or branched alkoxy, C1~C4 straight or branched alkylcarbonyl, C1~C4 straight or branched haloalkoxy, C1~C4 straight or branched alkylsulfonyl, C1~C4 straight or branched alkylsulfonylcarbamoyl, tetrazolyl, amino, phenyl, halophenyl, phenoxy and halophenoxy.

[0096] In another preferred embodiment, R2 is selected from methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyran-4-yl, 1-methylpiperidin-4-yl, 1-acetylpiperidin-4-yl, 1-methylsulfonylpiperidin-4-yl, 4-fluorobenzyl, phenyl, ethylcyclohexyl, and difluoro-substituted cyclohexyl;

[0097] R3, R4 and R5 are each independently selected from hydrogen and C1-C4 linear or branched alkyl.

[0098] In another preferred embodiment, R3, R4 and R5 are each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl and tert-butyl.

[0099] In another preferred embodiment, R3, R4 and R5 are each independently selected from hydrogen, methyl, ethyl, n-propyl and isopropyl.

[0100] In another preferred embodiment, a 1-(3-aminopropyl)-substituted cyclic amine compound represented by general formula (III), a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or mixture thereof is provided:

[0101] Wherein, the definitions of R1, R2, R3, R4, R5, R6 and W are as described in the general formula (I).

[0102] In the general formula (III), preferably, R1 is selected from the following groups which are unsubstituted or substituted with 1 to 3 substituents: The substituent is selected from halogen, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched halogenated alkyl, C1-C4 straight chain or branched alkylcarbonyloxy, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched halogenated alkoxy, NR 10 R 11 、-C(=O)R 12 , cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atom to which the substituents are attached form a 5-7 membered ring; preferably, the substituents are selected from halogen, C1-C2 alkyl, C1-C2 haloalkyl, C1-C2 alkoxy, C1-C2 alkylcarbonyloxy, C1-C2 haloalkoxy, NR 10 R 11 、-C(=O)R 12 , cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atom to which the substituents are attached constitute a 5-7 membered carbocyclic ring, a 5-7 membered heteroaromatic ring or a 5-7 membered heterocyclic ring; preferably, the substituents are selected from halogen, methyl, trifluoromethyl, trifluoromethoxy, methoxy, ethyl, amino, cyano, nitro, acetyl, formamido, acetamido, carbamoyl, N-methylcarbamoyl, N,N-dimethylcarbamoyl, formyloxy, acetoxy and methoxycarbonyl, or two adjacent substituents together with the carbon atom to which the substituents are attached constitute a benzene ring, a cyclopentene ring or a dioxole ring;

[0103] R 10and R 11 Each independently selected from H, C1-C4 linear or branched alkyl and -C(=O)R 13 Preferably, R 10 and R 11 Each independently selected from H, C1-C2 alkyl and -C(=O)R 13 ;

[0104] R 12 is selected from C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched alkoxy, hydroxy, amino (NH2) and C1-C4 straight chain or branched alkylamino; preferably, R 12 Selected from C1-C2 alkyl, C1-C2 alkoxy, hydroxy, amino (NH2) and C1-C2 alkylamino;

[0105] R 13 is selected from H and C1-C4 straight or branched alkyl; preferably, R 13 Selected from H and C1-C2 straight or branched chain alkyl;

[0106] R2 is selected from C1~C4 straight or branched alkyl and C3~C7 cycloalkyl which are unsubstituted or substituted with 1-3 substituents, wherein the substituents are selected from halogen, hydroxyl, C1~C4 straight or branched alkyl, C1~C4 straight or branched haloalkyl, C1~C4 straight or branched alkoxy, C1~C4 straight or branched alkylcarbonyl, C1~C4 straight or branched haloalkoxy, C1~C4 straight or branched alkylsulfonyl, C1~C4 straight or branched alkylsulfonylcarbamoyl, tetrazolyl, cyano and amino; preferably, R2 is selected from methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyran-4-yl, 1-methylpiperidin-4-yl, 1-acetylpiperidin-4-yl, 1-methylsulfonylpiperidin-4-yl, and difluoro-substituted cyclohexyl;

[0107] R3 and R4 are each independently selected from hydrogen and C1-C4 straight or branched alkyl; preferably, R3 and R4 are each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl and tert-butyl; preferably, R3 and R4 are each independently selected from hydrogen, methyl and ethyl;

[0108] R5 and R6 can be connected with Fragments together form

[0109] R7 is selected from hydrogen, C(=O)R8, C(=O)OR8, C(=O)NR8R9 and SO2R8; preferably, R7 is selected from C(=O)R8, C(=O)OR8 and SO2R8;

[0110] R8 and R9 are each independently selected from hydrogen and the following groups which are unsubstituted or substituted by 1-3 substituents: C1~C4 straight chain or branched alkyl, C1~C4 straight chain or branched haloalkyl, C3~C7 cycloalkyl and benzyl, wherein the substituents are selected from halogen, hydroxyl, C1~C4 straight chain or branched alkoxy, C1~C4 straight chain or branched alkyl, C1~C4 straight chain or branched haloalkyl, C1~C4 straight chain or branched haloalkoxy and amino, preferably selected from halogen, hydroxyl, methoxy, ethoxy, methyl, ethyl, trifluoromethyl, trifluoromethoxy and amino; preferably, R8 and R9 are each independently selected from hydrogen, C1~C4 straight chain or branched alkyl, C1~C4 straight chain or branched haloalkyl and C3~C7 cycloalkyl; preferably, R8 and R9 are each independently selected from methyl, ethyl, n-propyl, cyclopropyl, isopropyl, n-butyl, sec-butyl and tert-butyl.

[0111] In another preferred embodiment, the active ingredient is a compound selected from the following group, or a pharmaceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof:

[0112] Indications

[0113] Multiple sclerosis (MS) is a chronic, disabling, immune-mediated inflammatory demyelinating disease of the central nervous system that often affects the periventricular, juxtacortical, optic nerves, spinal cord, brainstem, and cerebellum. Lesions are characterized by spatial and temporal multiplicity. Spatial multiplicity refers to the presence of multiple lesions, with the cerebrum, brainstem, cerebellum, and spinal cord affected simultaneously or sequentially. Temporal multiplicity refers to a relapsing-remitting course. Due to the widespread involvement, repeated attacks can ultimately lead to disability or even death. Clinical manifestations vary, with common symptoms including decreased vision, diplopia, limb sensory and motor impairments, ataxia, and bladder or rectal dysfunction.

[0114] Relapsing-remitting multiple sclerosis (RRMS)

[0115] Relapsing-remitting multiple sclerosis is characterized by relapses, e.g., defined as episodes of new neurologic deficits or neurologic worsening lasting more than 24 hours, in the absence of fever or infection. There is no overt disease progression during remissions. At different time points, RRMS can be further characterized as active (with evidence of relapses and / or new MRI activity) or inactive, and worsening (with demonstrated increased disability within a specified time period following a relapse) or non-worsening.

[0116] Primary progressive multiple sclerosis (PPMS)

[0117] The term RMS (relapsing-relapsing multiple sclerosis) encompasses RRMS, secondary progressive multiple sclerosis (SPMS), and clinically isolated syndrome (CIS).

[0118] PPMS is characterized by worsening neurologic function (accumulation of disability) from the onset of symptoms, without early relapses or remissions. PPMS can be further characterized at different time points as active (with evidence of occasional relapses and / or new MRI activity) or inactive, and as progressive (evidence of objectively measured disease worsening over time, with or without relapses or new MRI activity) or nonprogressive. PPMS can have brief periods of stable disease, with or without relapses or new MRI activity, and periods when increasing disability occurs, with or without new relapses or MRI lesions.

[0119] Secondary progressive multiple sclerosis (SPMS)

[0120] SPMS develops after an initial relapsing-remitting course. Most people diagnosed with RRMS will eventually transition to a secondary progressive course in which progressive worsening of neurological function (accumulation of disability) occurs over time. SPMS can be further characterized as active (evidence of relapses and / or new MRI activity) or inactive at different time points, and as progressive (evidence of objectively measured worsening of the disease over time, with or without relapses) or non-progressive. In SPMS, occasional relapses may occur, as well as periods of stability.

[0121] Clinically isolated syndrome (CIS):

[0122] Clinically isolated syndrome (CIS) can refer to a single clinical episode of central nervous system (CNS) inflammatory demyelinating symptoms suggestive of multiple sclerosis (MS). CIS manifestations can be unifocal or multifocal and may typically involve the optic nerves, brainstem, cerebellum, spinal cord, or cerebral hemispheres.

[0123] The main advantages of the present invention include:

[0124] (a) The compounds of the present invention have excellent repairing effects on spinal cord demyelination in multiple sclerosis.

[0125] (b) The compounds of the present invention can significantly reduce the number of immune cells infiltrating the spinal cord and myeloid deviation in multiple sclerosis.

[0126] (c) The compounds of the present invention have a rapid onset of effect in treating multiple sclerosis and a significant effect in alleviating the symptoms of the disease.

[0127] (d) The compounds of the present invention can delay the progression of multiple sclerosis.

[0128] (e) The compounds of the present invention have good drugability and are suitable for long-term administration.

[0129] It should be understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, were generally performed under conventional conditions (e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989)) or as recommended by the manufacturer. Unless otherwise indicated, percentages and parts are by weight.

[0130] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0131] Example 1. Preparation of compound DC521022

[0132] The synthetic route of compound DC521022 is as follows:

[0133] The above route was used to synthesize compound 1-9. Compound 1-9 (35.6 mg, 0.1 mmol) was dissolved in 5 mL of dichloromethane, and triethylamine (28 μL, 0.2 mmol), 4,4-difluorocyclohexanecarboxylic acid (18 mg, 0.11 mmol), HOBt (14.8 mg, 0.11 mmol), and EDCI (21.1 mg, 0.11 mmol) were added in sequence. The mixture was stirred at room temperature and reacted for 12 hours. Water was added, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography (DCM:CH3OH=10:1) to obtain the final product, compound DC521022.

[0134] 1 HNMR (500MHz, CDCl3): δ7.31(dd,J=5.0,2.9Hz,1H),7.13(d,J=2.9Hz,1H),7.01(dd,J=4.9,1.4Hz,1H),4.29(tt,J=12.0,5.5 Hz,1H),3.41(s,2H),2.98(p,J=6.8Hz,1H),2.51(s,5H),2.37-1.97(m,10H),1.97-1.55(m,11H),1.38(dd,J=6.8,2.1Hz,6H). 13C NMR (125MHz, CDCl3) δ173.25,159.11,150.61,142.95,126.53,126.12,121.04,77.23,59.02,58.40,47.99 ,47.63,47.18,42.93,35.52,35.34,34.51,33.02,32.82,26.72,26.03,25.96,25.86,21.67,13.21.ESI-MS m / z:520.3[M+H] + .HRMS(ESI)calcd for C 27 H 39 F2N5OS([M+H] + ):520.2916;found:520.2917.

[0135] Example 2. In vivo multiple sclerosis onset test

[0136] Autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis. In this experiment, behavioral scores and body weight assessments were performed on two groups of mice every day until the end of the experiment to investigate the effect of compound DC521022 on the pathogenesis of multiple sclerosis.

[0137] Experimental animals: 10-week-old female C57 / BL6 mice were randomly divided into experimental and control groups: model control group (EAE+Vehicle): administered with saline after model establishment; model drug group (EAE+DC521022): administered with DC521022 suspension after model establishment. Based on the timing and method of administration, the model control group and the model drug group were divided into four groups: intraperitoneal injection of 50 mg / kg DC521022 after immunization, oral administration of 50 mg / kg DC521022 after immunization, intraperitoneal injection of 50 mg / kg DC521022 at the onset of symptoms, and oral administration of 50 mg / kg DC521022 at the onset of symptoms. The control group received the same dose of saline at the same time.

[0138] Preparation method of DC521022: Weigh an appropriate amount of DC521022 according to the preparation plan, first add an appropriate amount of solvent and place it in a beaker marked with a certain volume, then add an appropriate amount of hydrochloric acid (hydrochloric acid dosage calculation formula: hydrochloric acid: test sample = 1:1 (molar ratio)) into the beaker, slowly add the test sample, and then shake rapidly and continuously at room temperature for about 16 hours until the test sample is completely dissolved and the solution is clear, and then add solvent to the corresponding scale line.

[0139] Animal Experimental Methods: Mouse polypeptide antigen MOG35-55 (purity >95%) and complete Freund's adjuvant containing inactivated Mycobacterium tuberculosis were repeatedly mixed in a syringe to emulsify the mixture into a water-in-oil solution. The emulsion (200 μg MOG35-55 (CAS No. 149635-73-4) and 500 μg inactivated Mycobacterium tuberculosis) was injected subcutaneously into the skin at the midpoint of the line connecting the upper and lower limbs of the mice, with 100 μL injected into each subcutaneous injection for a total of 200 μL. Pertussis toxin (200 ng) was injected intraperitoneally on the day of immunization and two days later. Following the completion of the animal experiment, the mice were observed daily, and clinical scores were recorded. The scale ranged from 0 to 5: 0: no symptoms; 1: tail weakness; 2: hindlimb weakness or unsteady gait; 3: complete hindlimb paralysis; 4: complete hindlimb paralysis with forelimb weakness or paralysis; 5: death.

[0140] Explanation of successful modeling: After the above modeling, the mice showed tail drooping symptoms, indicating that the mice were successfully immunized with MOG35-55 to induce autoimmune encephalomyelitis (EAE).

[0141] Administration method: Model group (EAE+DC521022): EAE was induced in C57 / BL6 mice by MOG35-55 immunization, and 50 mg / kg of DC521022 was injected intraperitoneally and perfused on the day of modeling and the first day of onset, respectively; Model group (EAE+Vehicle): EAE was induced in C57 / BL6 mice by MOG35-55 immunization, and the same dose of normal saline was injected intraperitoneally and perfused on the day of modeling and the first day of onset, respectively.

[0142] Experimental Methods: Following model establishment, DC521022 suspension was administered immediately via intraperitoneal injection and perfusion on the day of immunization and the first day of disease onset, respectively. A control group of mice received normal saline. Dosing was repeated daily, and the mice were observed and clinically scored until days 14 and 28 after model establishment. At the peak of EAE and two weeks after treatment, bone marrow analysis was performed for myeloid lineage deviation, spinal cord demyelination, and inflammatory cell infiltration in the spinal cord.

[0143] Experimental results showed that mice treated with DC521022 developed disease later than the control group and had lower scores. DC521022 treatment also improved the mice's weight, with treated mice showing increased weight compared to the control group (Figure 1). This class of compounds holds great promise for the treatment of multiple sclerosis and therefore has significant commercial value.

[0144] Example 3. In vivo testing of spinal cord demyelination levels in multiple sclerosis

[0145] In this study, immunohistochemistry was used to perform fast blue staining of the mouse spinal cord to compare the levels of spinal cord demyelination in the DC521022-treated group and the saline-treated group 14 and 28 days after the successful EAE model.

[0146] Preparation of paraffin sections: On the first day after the successful induction of the EAE model, the mice were divided into experimental and control groups. Two of the experimental groups were injected with DC521022 intraperitoneally or orally on the day of modeling, and the other two groups were injected with DC521022 intraperitoneally or orally at the onset of the disease in EAE mice. The control group was injected with the same dose of normal saline in the same way as the control group. The mice were observed for 2 weeks for those who received the drug on the day of immunization, and the mice were observed for 2 weeks for those who received the drug at the onset of symptoms. After 2 weeks of treatment, the mice were anesthetized and the spinal cords were fixed after perfusion with phosphate buffer and 4% paraformaldehyde. First, the mouse was deeply anesthetized with 20 μg / g tribromoethanol and properly fixed. The skin was cut longitudinally along the chest-abdomen line to expose the chest and abdominal cavities, and then the heart was freed and the liver was cut. The venous access was opened, and 20 mL of PBS and 20 mL of 4% paraformaldehyde were perfused through the left ventricle in sequence. The standard of successful perfusion was that the liver and other abdominal organs turned white. After the perfusion was completed, the back was cut open along the midline, the spine and spinal cord were peeled off, from the thoracic vertebrae to the caudal vertebrae, and then cut open along one side of the spine to expose the spinal cord. The lumbar cord segment was taken and placed in 4% paraformaldehyde and immersed at 4°C for more than 16 hours. After fixation, the brain tissue was removed from the formaldehyde, placed in a labeled tissue embedding cassette, and then placed in 50% ethanol, 75% ethanol, 85% ethanol, 95% ethanol, and anhydrous ethanol (twice) for 1 hour each, xylene for 20 minutes (twice), soft wax (52-54°C), and hard wax (56-58°C) for 1.5 hours each. The specimens were dehydrated, transparentized, and immersed in wax in the above solutions before embedding. After embedding, the spinal cord tissue of the experimental and control groups was removed and cut into 5-μm thick sections using a paraffin microtome. After complete expansion in 40°C distilled water, the sections were removed and dried in a 60°C slide dryer for 2 hours before use. The paraffin sections were stored at room temperature.

[0147] Immunohistochemical staining procedures: Spinal cord sections were sequentially exposed to xylene for 5 min (twice), anhydrous ethanol for 5 min (twice), 95% ethanol, 85% ethanol, 75% ethanol, 50% ethanol, and ultra-light water (DDW) for 2 min each. The sections were then placed in a staining box containing Luxol Fast Blue Solution and incubated at 60°C for 2 h. The sections were then rinsed thoroughly with distilled water for 5 min and immersed in lithium carbonate multiple times, each for no more than 20 seconds. The sections were then repeatedly exposed to alcohol reagent and observed under a microscope while rinsing until the gray matter was colorless and the white matter remained blue. The sections were then rinsed again with distilled water and placed in Cresyl Echt Violet reagent for 2–5 min. After a quick rinse in distilled water, the sections were rapidly dehydrated in anhydrous ethanol. Finally, the sections were mounted with synthetic resin and photographed as quickly as possible under a standard microscope. Images were analyzed using ImageJ software. The demyelinated and white matter areas were averaged across six sections of each lumbar spinal cord segment to calculate the percentage of demyelinated area to total area.

[0148] The results showed that after 14 and 28 days of administration, the demyelination of mice treated with DC521022 was improved compared with the control group, and it was able to have a significant repair effect on spinal cord demyelination (as shown in Figure 2).

[0149] Example 4. In vivo testing of immune cell levels in the spinal cord of multiple sclerosis

[0150] In this experiment, immunohistochemistry was used to perform HE staining of the mouse spinal cord to compare the levels of infiltrating immune cells in the spinal cord of the DC521022-treated group and the saline-treated group 14 and 28 days after the successful establishment of the EAE model.

[0151] The steps for preparing paraffin sections were the same as those for fast blue staining.

[0152] Immunohistochemical staining procedures: Spinal cord sections were sequentially placed in xylene for 5 min (twice), anhydrous ethanol for 5 min (twice), 95% ethanol, 85% ethanol, 75% ethanol, 50% ethanol, and DDW for 2 min each. Stain with hematoxylin solution for 5 min and rinse with tap water for 5 s. Differentiation solution was used for 3 s, followed by a 20 s rinse with tap water to remove the differentiation solution. Bluing solution was used for 30 s, followed by a 20 s rinse with tap water to remove the blueing solution. Stain with eosin for 30 s, rinse with tap water for 5 s, dehydrate, and mount.

[0153] The images were analyzed using imageJ software. The inflammatory cell infiltration and white matter area at six levels of each lumbar spinal cord segment were averaged to calculate the percentage of inflammatory cell infiltration area to total area.

[0154] The experimental results showed that after long-term treatment with DC521022, the number of immune cells infiltrating the spinal cord of mice decreased, with statistical significance (as shown in Figure 3).

[0155] Example 5. In vivo testing of myeloid bias in multiple sclerosis

[0156] This study used flow cytometry to investigate the myeloid deviation of the compound DC521022 in the bone marrow of each group of mice in this model on the 14th and 28th days of the experiment.

[0157] To prepare a single-cell suspension of brain tissue: deeply anesthetize mice with intraperitoneal injection of tribromoethanol and properly fix them. Remove the left femur of the mouse and soak it in ice-cold PBS for later use. Aspirate the bone marrow from the femur into a 15 mL centrifuge tube and centrifuge at 350g for 5 minutes. Discard the supernatant and add 1 mL of red blood cell lysis buffer. After lysis for 6 minutes, add 5 mL of PBS to terminate lysis and centrifuge at 350g for 5 minutes. Resuspend in 1 mL of PBS and count the cells.

[0158] Flow cytometry antibody staining The following fluorescent dye channels were selected for flow cytometry antibody staining: fluorescein isothiocyanate (FITC), phycoerythrin (PE), perdinoflagellates chlorophyll protein (PerCP-Cy5.5), allophycocyanin (APC), allophycocyanin-anthocyanin 7 (APC-Cy7), and phycoerythrin-anthocyanin 7 (PE-Cy7).

[0159] Specific flow cytometry antibodies against mouse target antigens: anti-mouse lin- antibody, anti-mouse sca-1 antibody, anti-mouse ckit antibody, anti-mouse CD48 antibody, anti-mouse CD150 antibody, anti-mouse CD127 antibody, anti-mouse CD135 antibody, anti-mouse CD34 antibody, anti-mouse 16 / 32 antibody, anti-mouse CD115 antibody, anti-mouse 7CD135 antibody.

[0160] Surface antibody staining: Add surface antibody to a 100 μL single-cell suspension in a flow cytometry tube and vortex to mix thoroughly (add in the correct dosage according to the instructions). Incubate at 4°C in the dark for 30 minutes. Wash with 1 mL of PBS, centrifuge at 1500 rpm for 5 minutes, discard the supernatant, resuspend in 400 μL of PBS, vortex to mix thoroughly, and filter.

[0161] In this experiment, flow cytometry antibody staining used blank controls, single positive compensation, isotype controls, and fluorescence minus one (FMO) controls. Flow cytometry data were collected using a FACS Aria II and analyzed using FlowJo V10 (Version 10, FlowJo, LLC).

[0162] The experimental results showed that on days 14 and 28, the proportion and number of hematopoietic stem cells in the bone marrow of EAE mice treated with DC521022 were significantly reduced, and the deviation toward the myeloid lineage was significantly reduced (as shown in Figure 4).

[0163] In summary, long-term administration of the test substance DC521022 can delay the onset of multiple sclerosis in mice in the EAE model, has a significant repair effect on demyelination in mice, and reduces the number of immune cells infiltrating the spinal cord and myeloid deviation.

[0164] Compared with the reported effects of the selective CCR5 antagonist DAPTA (see Figure 5 or Sheikh et al., Cellular Immunology. 2022, 379, 104580, Figure 1), administration of the drug after the onset of disease in mice on the 6th day did not delay the onset of the disease in mice, and even aggravated the progression of the disease. The therapeutic effect was not gradually manifested until the 22nd day of administration.

[0165] The therapeutic efficacy of the present invention in EAE mice was significantly superior to that of the studies conducted by Sheikh et al. Two drug-administered models were used. When administered on the day of modeling, EAE mice developed significantly later than the control group, with lower behavioral scores and better body weight (Figures 1A and 1C). When administered after the onset of symptoms, disease progression was significantly delayed. Furthermore, behavioral scores were lower and body weight was significantly improved (Figures 1B and 1D).

[0166] All documents mentioned herein are incorporated herein by reference as if each document were individually incorporated by reference. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the appended claims.

Claims

1. A use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof, characterized in that: For preparing a drug for treating and / or preventing multiple sclerosis and related diseases; The compound of formula (I) has the following structure: in, W does not exist or is -CH2CH2-; X is N or CR6; R1 is selected from a 5-7 membered heteroaryl group which is unsubstituted or substituted by 1-3 substituents, wherein the heteroaryl group contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen, and the substituents are each independently selected from halogen, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched haloalkoxy, -NR 10 R 11 、-C(=O)R 12 , C1-C4 straight or branched chain alkanoyloxy, cyano, nitro and hydroxyl, or two adjacent substituents together with the carbon atom to which they are attached form a 5-7 membered ring; R 10 and R 11 Each independently selected from H, C1-C4 straight or branched alkyl and -C(=O)R 13 ; R 12 Selected from C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched alkoxy, hydroxy, amino (NH2) and C1-C4 straight chain or branched alkylamino; R 13 Selected from H and C1-C4 straight or branched chain alkyl; R2 is selected from the following groups which are unsubstituted or substituted by 1-3 substituents: C1-C6 straight chain or branched alkyl, C3-C7 cycloalkyl, 4-7 membered heterocyclyl, C6-C12 aryl or 5-7 membered heteroaryl; wherein the substituent is selected from halogen, hydroxyl, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched alkylcarbonyl, C1-C4 straight chain or branched haloalkoxy, C1-C4 straight chain or branched alkylsulfonyl, C1-C4 straight chain or branched alkylsulfonylcarbamoyl, tetrazolyl, cyano, nitro, amino, carboxyl, phenyl and phenoxy; R3, R4 and R5 are each independently selected from hydrogen, C1-C6 straight or branched alkyl, C1-C6 straight or branched halogenated alkyl and C3-C7 cycloalkyl; R6 is selected from hydrogen and C1-C6 straight or branched alkyl; Alternatively, R5 and R6 can be connected with Together we form R7 is selected from hydrogen, C(=O)R8, C(=O)OR8, C(=O)NR8R9, SO2R8 and the following groups substituted by 1-3 substituents: C1-C6 straight chain or branched alkyl, C3-C7 cycloalkyl, 4-7 membered heterocyclyl, benzyl, C6-C12 aryl and 5-7 membered heteroaryl; wherein the substituent is selected from halogen, hydroxyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched haloalkoxy, cyano, nitro, amino, carboxyl; R8 and R9 are each independently selected from the following groups which are hydrogen, unsubstituted or substituted by 1-3 substituents: C1-C6 straight chain or branched alkyl, C3-C7 cycloalkyl, 4-7 membered heterocyclyl, benzyl, C6-C12 aryl and 5-7 membered heteroaryl; wherein the substituents are selected from halogen, hydroxyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched haloalkyl, C1-C4 straight chain or branched haloalkoxy, cyano, nitro, amino, and carboxyl.

2. The use according to claim 1, characterized in that The multiple sclerosis is selected from the group consisting of relapsing-remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, progressive relapsing multiple sclerosis, or a combination thereof.

3. The use according to claim 1, characterized in that The related diseases are selected from the following group: neuromyelitis optica, acute disseminated encephalomyelitis, stroke, craniocerebral injury, epilepsy, Alzheimer's disease, Parkinson's disease, immune inflammatory response, and demyelinating injury.

4. The use according to claim 1, characterized in that The multiple sclerosis and related diseases are selected from the following group: relapsing-remitting multiple sclerosis, secondary progressive multiple sclerosis, primary progressive multiple sclerosis, stroke, craniocerebral injury, epilepsy, immune inflammatory response, and demyelinating injury.

5. The use according to claim 1, characterized in that The compounds are used for: (1) Repair demyelination in the spinal cord in multiple sclerosis; (2) reduce the number of immune cells infiltrating the spinal cord in multiple sclerosis; (3) Reduce myeloid deviation in the spinal cord in multiple sclerosis.

6. The use according to claim 1, characterized in that The compound of formula (I) is selected from the following group:

7. The use according to claims 1 to 6, characterized in that The administration route of the compound of formula (I) is selected from the group consisting of oral, rectal, and parenteral.

8. The use according to claim 7, characterized in that The parenteral administration method is selected from the group consisting of intravenous, intramuscular or subcutaneous administration.

9. The use according to claim 7, characterized in that The oral preparation is selected from the group consisting of capsules, tablets, pills, powders, granules, emulsions, solutions, suspensions, syrups and tinctures.

10. A use of a pharmaceutical composition, characterized in that: The pharmaceutical composition is used to prepare drugs for treating and / or preventing multiple sclerosis and related diseases; And the pharmaceutical composition comprises: (i) a first active ingredient, wherein the first active ingredient is selected from the following group: a compound of formula (I), or a pharmaceutically acceptable salt, solvate, optically pure isomer, or stereoisomer thereof; (ii) optionally, a second active ingredient selected from the group consisting of interferon beta-1a, interferon beta-1b, teriflunomide, fingolimod, copaxone, ofatuzumab, or a combination thereof; (iii) a pharmaceutically acceptable carrier; The compound of formula (I) is as defined in claim 1.

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