Application of MAT2A inhibitor in preparation of medicine for treating hyperhomocysteinemia

Abstract

The invention relates to the field of biological medicines, in particular to application of an MAT2A inhibitor in preparation of a medicine for treating hyperhomocysteinemia. The MAT2A inhibitor plays an important role in homocysteine regeneration in a metabolism remethylation pathway of homocysteine, by using the MAT2A inhibitor, the homocysteine regeneration can be blocked, the content of the homocysteine is effectively reduced, and methionine serving as essential amino acid can be metabolized through other pathways, so that the homocysteine metabolism remethylation pathway of the homocysteine can be promoted, and the homocysteine metabolism remethylation pathway of the homocysteine can be promoted. No toxic or side effect is caused on a using object.

Description

TECHNICAL FIELD

[0001] The present specification relates to the field of biological medicine, in particular to the application of MAT2A inhibitors in the preparation of drugs for treating hyperhomocysteinemia. BACKGROUND

[0002] Hyperhomocysteinemia is a disease characterized by elevated homocysteine in the blood, usually referring to homocysteine > 15 μmol / L. Hyperhomocysteinemia is an independent risk factor for cardiovascular disease, cerebrovascular disease, cognitive impairment, and osteoporotic fractures, with a global prevalence of 4.2% and a national prevalence of 27.5%.

[0003] Studies have found that unhealthy diets such as heavy drinking, smoking, and heavy coffee drinking can lead to elevated plasma homocysteine levels. In addition, plasma homocysteine concentration increases with age; the plasma homocysteine concentration in men is higher than that in women.

[0004] Homocystinuria is a rare autosomal recessive genetic disease, which is a congenital sulfur metabolism error caused by homocysteine metabolism disease. The classic homocystinuria is caused by the down-regulation of cystathionine-β-synthase (CBS) enzyme activity, which is the loss of function of the enzyme in the trans-sulfur pathway to convert homocysteine to cystathionine, resulting in elevated levels of methionine and homocysteine in the blood and urine. The natural course of homocystinuria caused by CBS deficiency is well characterized, indicating that the destructive complications of the disease are closely related to long-term exposure to elevated plasma total homocysteine (tHcy) levels. Previous studies have shown that reducing plasma tHcy has a consistent and beneficial effect on reducing disease-related complications. Therefore, the treatment guidelines defined by the patent emphasize that the primary treatment goal for homocystinuria caused by CBS deficiency is to reduce plasma tHcy.

[0005] It is known that patients with homocystinuria caused by CBS deficiency will suffer from a series of metabolic diseases, including eye complications (high myopia, retinal degeneration, secondary glaucoma, etc.), cardiovascular diseases (thromboembolism, atherosclerosis), skeletal system diseases (scoliosis, pectus excavatum, etc.), liver damage, central nervous system diseases (mental retardation), and developmental retardation. Therefore, for subjects with homocystinuria caused by CBS deficiency, management of adaptive behavior can be clinically relevant, and it is reasonable to assume that early treatment to reduce plasma tHcy can lead to improvement in adaptive behavior.

[0006] Methionine adenosyltransferase 2a (MAT2A) plays an important role in metabolism and epigenetics. It is currently known that MAT2A is involved in methionine metabolism, as a member of the methionine adenosyltransferase family, mainly responsible for catalyzing the reaction of methionine with ATP to generate S-adenosyl methionine (SAM) and AMP, as the first step of methionine metabolism, which is essential for maintaining the normal level of methionine in the body. Secondly, MAT2A affects DNA methylation, SAM is an important methyl donor in the process of DNA methylation, and the activity of MAT2A directly affects the methylation state of DNA, which may affect the expression and regulation of genes. MAT2A inhibitors can provide new therapies for cancer patients, including cancer patients with MTAP-deficient tumors, but no studies have found effects on homocysteine elevation and its complications.

[0007] Current disease management for subjects with hyperhomocysteinuria due to CBS deficiency includes restricting dietary protein and methionine with the goal of lowering plasma tHcy concentration to recommended levels while maintaining adequate nutrition. Frequent metabolic monitoring by plasma amino acid analysis is required to assess response to the diet. In addition, compliance is poor due to the difficulty in maintaining the degree of protein restriction in the diet for subjects with hyperhomocysteinuria due to CBS deficiency. Furthermore, this degree of dietary protein restriction can lead to long-term nutritional deficiencies, resulting in non-compliance and overall disease burden. It is challenging for many subjects with hyperhomocysteinuria to adhere to a strict amino acid showing diet. Therapeutic options that allow patients to relax dietary restrictions would improve patient quality of life. Therefore, there is still a need to seek methods that can regulate plasma tHcy concentration on the basis of not restricting the diet, minimize the level of toxic plasma tHcy, and reduce the serious consequences of hyperhomocysteinuria. SUMMARY

[0008] To solve the above technical problems, the purpose of the present application is to provide the use of MAT2A inhibitors in the preparation of a drug for treating hyperhomocysteinemia, the present application finds that the MAT2A target of the hyperhomocysteinemia model is significantly increased through transcriptomic data, based on this target, it is predicted that MAT2A inhibitors may be effective drugs for hyperhomocysteinemia.

[0009] To achieve the above object and other related objects, the present application provides the use of MAT2A inhibitors or their pharmaceutically acceptable salts, esters, isomers, prodrugs, polymorphs or solvates in the preparation of a drug for treating hyperhomocysteinemia, as a new possible effective drug for hyperhomocysteinemia patients, which can reduce the content of homocysteine in the blood of hyperhomocysteinemia patients.

[0010] The first aspect of the present application provides a use of a MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof in the preparation of a medicament for treating hyperhomocysteinemia or its complications.

[0011] The MAT2A inhibitor refers to a substance capable of reducing the expression of MAT2A and / or hindering the function thereof, which can be any substance capable of reducing the expression of MAT2A and / or hindering the function of MAT2A in the prior art.

[0012] In some embodiments of the present application, the MAT2A inhibitor includes AG-270, IDE-397. The AG-270 is a MAT2A inhibitor known to those skilled in the art, with a molecular formula of C30H27N5O2, a molecular weight of 489.6, a CAS number of 2201056-66-6, and a structural formula as shown in Formula I below:

[0013]

[0014] The IDE-397 is a MAT2A inhibitor known to those skilled in the art, with a molecular formula of C 14 H8ClF3N4O, a molecular weight of 340.69, a CAS number of 2439277-80-0, and a structural formula as shown in Formula II below:

[0015]

[0016] The second aspect of the present application provides a composition for treating hyperhomocysteinemia, which comprises the MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof as described in the use above, and a pharmaceutically acceptable carrier, medium.

[0017] In a preferred embodiment, the composition includes a pharmaceutical composition, a food composition or a health product composition.

[0018] In a preferred embodiment, the dosage form of the composition is selected from the group consisting of an injection, a sterile powder for injection, a tablet, a capsule, a liquor, a powder, a granule, a syrup, a solution, a tincture, an aerosol, a powder mist, or a suppository.

[0019] The third aspect of the present application provides a use of the above-mentioned pharmaceutical composition in the preparation of a medicament for treating hyperhomocysteinemia or its complications.

[0020] The beneficial effects brought by the present application include but are not limited to:

[0021] The MAT2A inhibitor of the present application plays an important role in the remethylation pathway of homocysteine metabolism, and by using the MAT2A inhibitor, the homocysteine regeneration can be blocked, the content of homocysteine can be effectively reduced, and methionine as an essential amino acid can be metabolized through other pathways, and has no toxic side effects on the use object. The present application expands the application range of the MAT2A inhibitor, and the MAT2A inhibitor can be further developed as a new drug for reducing the concentration of blood homocysteine. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 Effect of the MAT2A inhibitor of Example 1 on homocysteine of cbsb deletion zebrafish is shown;

[0023] Figure 2 Effect of the cbsb deletion zebrafish of Example 1 on eyes and lenses is shown;

[0024] Figure 3 Effect of the MAT2A inhibitor of Example 1 on eyes and lenses of cbsb deletion zebrafish is shown;

[0025] Figure 4 Detection chart of the case of intravenous embolism of the cbsb deletion zebrafish of Example 1 is shown;

[0026] Figure 5 Effect of the MAT2A inhibitor of Example 1 on the heart of cbsb deletion zebrafish is shown;

[0027] Figure 6 Detection chart of the case of calcium absorption of the cbsb deletion zebrafish of Example 1 is shown;

[0028] Figure 7 Effect of the MAT2A inhibitor of Example 1 on the calcium absorption capacity of cbsb deletion zebrafish is shown;

[0029] Figure 8 Treatment effect of the MAT2A inhibitor of Example 2 on cbs deletion mice is shown;

[0030] Figure 9 Treatment effect of the MAT2A inhibitor of Example 3 on human hyperhomocysteinemia is shown;

[0031] Figure 10 Treatment effect of the MAT2A inhibitor of Example 3 on human liver homocysteinemia is shown;

[0032] Figure 11 Treatment effect of the MAT2A inhibitor of Example 4 on methionine-induced hyperhomocysteinemia is shown. DETAILED DESCRIPTION

[0033] As shown in the specification and claims herein, unless the context clearly indicates otherwise, the words "comprise", "comprising", "consisting of" and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense that may be preferred by some, and the articles "a", "an", and "one" are not intended to be construed as excluding the presence of zero or more of the referenced elements.

[0034] The present application first discovers that the MAT2A inhibitor can effectively reduce the content of homocysteine in blood by inhibiting the generation of methionine adenosyltransferase and further blocking the regeneration of homocysteine, and has excellent effect on treating hyperhomocysteinemia.

[0035] The present application first provides the use of the MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof in the preparation of a drug for treating hyperhomocysteinemia or its complications.

[0036] In the present application, the hyperhomocysteinemia includes diseases in which the content of homocysteine in blood is significantly increased.

[0037] In the present application, the complications are selected from one or more of the following: eye complications, cardiovascular diseases, skeletal system diseases, liver damage, central nervous system diseases, and developmental retardation.

[0038] In the specific embodiments of the present application, the eye complications are manifested as high myopia, retinal degeneration, and secondary glaucoma.

[0039] In the specific embodiments of the present application, the cardiovascular diseases are manifested as thromboembolism and atherosclerosis.

[0040] In the specific embodiments of the present application, the skeletal system diseases are manifested as osteoporosis, scoliosis, and pectus excavatum.

[0041] In the specific embodiments of the present application, the central nervous system diseases are manifested as mental retardation and cognitive impairment.

[0042] In some embodiments of the present application, the content of homocysteine in a patient with hyperhomocysteinemia is higher than that in a healthy population, and after the MAT2A inhibitor is used in the patient, the content of homocysteine in the patient is lower than that when the MAT2A inhibitor is not used.

[0043] In the present application, the MAT2A inhibitor refers to a substance that can reduce the expression of MAT2A and / or hinder the function thereof. It can be any substance in the prior art that can reduce the expression of MAT2A and / or hinder the function of MAT2A.

[0044] In some embodiments, the MAT2A inhibitor has at least one of the following effects:

[0045] (1) alleviating liver inflammation;

[0046] (2) alleviating lens shrinkage;

[0047] (3) improving heart function;

[0048] (4) improving vascular wall thickening;

[0049] (5) improving calcium absorption capacity.

[0050] In certain embodiments of the present application, the MAT2A inhibitor includes AG-270, IDE-397. The AG-270 is a MAT2A inhibitor known to those skilled in the art, with a molecular formula of C30H27N5O2, a molecular weight of 489.6, a CAS number of: 2201056-66-6, and a structural formula as shown in Formula I below:

[0051]

[0052] The IDE-397 is a MAT2A inhibitor known to those skilled in the art, with a molecular formula of C 14 H8ClF3N4O, a molecular weight of 340.69, a CAS number of: 2439277-80-0, and a structural formula as shown in Formula II below:

[0053]

[0054] In certain embodiments of the present application, the salts, esters of the MAT2A inhibitors can be used in the form of pharmaceutically or physiologically acceptable salts or esters. Reference to "pharmaceutically acceptable salts" typically refers to any salt (typically meaning nontoxic, particularly as a result of the counterion) that is physiologically tolerable when properly administered to treat (particularly in humans and / or mammals). These physiologically tolerable salts can be formed with cations or bases, and in the context of the present application, particularly when administered in humans and / or mammals, they should be understood to be salts formed from at least one compound provided according to the present application, typically the acid (deprotonated), such as an anion, and at least one physiologically tolerable cation (preferably an inorganic cation). In the context of the present application, this can specifically include salts with alkali metals and alkaline earth metals, as well as with ammonium cations (NH4+), specifically can include but are not limited to salts with (mono) or (di)sodium, (mono) or (di)potassium, magnesium, or calcium. These physiologically tolerable salts can also be formed with anions or acids, and in the context of the present application, particularly when administered in humans and / or mammals, they should be understood to be salts formed from at least one compound provided according to the present application, typically the protonated, such as a cation, and at least one physiologically tolerable anion. The salts, esters of the MAT2A inhibitors include but are not limited to salts or esters with hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, succinic acid, oxalic acid, fumaric acid, maleic acid, oxalacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or isethionic acid. Salts of halides are also applicable. Other salts include those with alkali metals or alkaline earth metals (such as sodium, potassium, calcium, or magnesium).

[0055] In certain embodiments of the present application, the "prodrug" refers to a "prodrug" that, when administered in an appropriate manner, is metabolized or chemically reacts in the human body to transform into the active MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph, or solvate thereof.

[0056] In certain embodiments of the present application, the MAT2A inhibitors can also be used in conjunction with other therapeutic agents. When the active MAT2A inhibitors are used in conjunction with other therapeutic agents, the MAT2A inhibitors and other therapeutic agents are co-administered. "Co-administered" means administration simultaneously in the same formulation or in two different formulations via the same or different routes, or sequentially via the same or different routes. "Sequentially" administered means that there is a time difference between the administration of two or more different compounds in seconds, minutes, hours, or days.

[0057] The present application also provides a composition for treating hyperhomocysteinemia, which comprises the MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof as described above in the use, and optionally one or more pharmaceutically acceptable adjuvants. The pharmaceutically acceptable adjuvants are selected from stabilizers, excipients, antioxidants (ascorbic acid, etc.), buffers (phosphoric acid, citric acid, other organic acids, etc.), preservatives, surfactants (PEG, Tween, etc.), chelating agents (EDTA, etc.), binders, etc. Furthermore, other low-molecular-weight polypeptides; serum albumin, gelatin or immunoglobulin, etc. proteins; glycine, glutamine, asparagine, arginine and lysine, etc. amino acids; polysaccharides and monosaccharides, etc. sugars or carbohydrates; mannitol or sorbitol, etc. sugar alcohols can also be contained. When an aqueous solution for injection is prepared, for example, a physiological saline, an isotonic solution containing glucose or other auxiliary drugs, such as D-sorbitol, D-mannose, D-mannitol, sodium chloride, can be used with a suitable solubilizing agent such as an alcohol (ethanol, etc.), a polyol (propylene glycol, PEG, etc.), a nonionic surfactant (Tween 80, HCO 50), etc.

[0058] The composition of the present application includes, but is not limited to, a pharmaceutical composition, a food composition, and a health product composition.

[0059] In the pharmaceutical composition of the present application, the MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof can be a single active ingredient or can be combined with other active ingredients to form a combined preparation.

[0060] In some embodiments of the present application, the content of the active ingredient (MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof) is generally a safe and effective amount, which should be adjustable for those skilled in the art, for example, the amount of the active ingredient administered is generally dependent on the body weight of the patient, the type of application, the condition and severity of the disease, for example, the amount of the active ingredient administered can be generally 10-200 mg / Kg of body weight.

[0061] The effective amount administered can be considered by those skilled in the art according to the severity of the condition and the health and age of the recipient. The effective amount can generally vary between 0.01 ng / Kg of body weight and about 100 mg / Kg of body weight.

[0062] The active ingredient or pharmaceutical composition containing the active ingredient provided by the present application can be adapted to any form of administration, and can be administered orally or parenterally, for example, can be administered by the lung, nose, rectum and / or intravenous injection, more specifically can be administered intradermally, subcutaneously, intramuscularly, intraarticularly, intraperitoneally, by the lung, orally, sublingually, nasally, transdermally, vaginally, orally or parenterally; injection administration includes intravenous injection, intramuscular injection and subcutaneous injection, etc., transdermal administration, etc.

[0063] As used herein, the dosage form of the pharmaceutical composition is selected from the group consisting of injection, sterile powder for injection, tablet, pill, capsule, lozenge, elixir, powder, granule, syrup, solution, tincture, aerosol, powder mist, or suppository. Those skilled in the art can select a suitable preparation form according to the administration mode, for example, the preparation form suitable for oral administration can be, but is not limited to, pill, tablet, chewable agent, capsule, granule, solution, drop, syrup, aerosol or powder mist, etc., and further, for example, the preparation form suitable for parenteral administration can be, but is not limited to, solution, suspension, rehydratable dry preparation or spray, etc., and further, for example, the preparation form suitable for rectal administration can be suppository, and further, for example, the preparation form suitable for injection administration can be injection, sterile powder for injection, etc.

[0064] Among them, tablets, lozenges, pills, capsules, etc. can also contain the following components: binding agents, such as gum, acacia, corn starch or gelatin; excipients, such as dicalcium phosphate; disintegrating agents, such as corn starch, potato starch, alginic acid, etc.; lubricants, such as magnesium stearate; sweeteners, such as sucrose, lactose or saccharin, or flavoring agents, such as peppermint, oil of wintergreen or cherry flavoring, can be added. When the unit dosage form is a capsule, in addition to the above-mentioned substances, it can contain a liquid carrier. Various other substances can be present in the form of coatings or to improve the physical form of the unit dosage form. For example, tablets, pills or capsules can be coated with shellac, sugar or both. Syrups or elixirs can contain the active compound, sucrose as a sweetener, methyl and propyl parabens as preservatives, colorants and flavoring agents, such as cherry or orange flavoring agents. Any substance used to prepare any unit dosage form should be pharmaceutically pure and essentially non-toxic in the amount used. In addition, the active compound can be incorporated into a sustained release preparation or formulation.

[0065] The present application also provides the use of the aforementioned composition in the preparation of a product for treating hyperhomocysteinemia or its complications.

[0066] The present application is illustrated by the following non-limiting examples. Other advantages and benefits of the present application will become apparent to those skilled in the art, upon all the attached claims and appended drawings, and upon reading the foregoing description of the application. The present application can be implemented or carried out in other ways than those specifically described herein without departing from the spirit of the present application. Many variations and modifications of the embodiments disclosed herein can be made which fall within the scope of the application. Any feature or subcombination of features disclosed herein can be provided with or without its combination with other features or subcombinations thereof.

[0067] Before further describing the present application, it is to be understood that the application is not limited to the particular specific embodiments described herein; it is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting as to the scope of the present application. In this specification and in the claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

[0068] When numerical ranges are given, it should be understood that every numerical range encompasses any number falling within the range. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains and any method and material similar or equivalent in function, in so far as relevant to the present application, are also intended to be encompassed by the present application.

[0069] The experimental methods in the following examples are routine unless otherwise specified. The experimental materials used in the following examples are purchased from routine biochemical reagent companies unless otherwise specified. The quantitative experiments in the following examples are set up in triplicate, and the results are averaged.

[0070] Example 1 - Therapeutic effect of MAT2A inhibitors on zebrafish experimental model of hyperhomocysteinemia

[0071] 1. Zebrafish breeding

[0072] AB strain zebrafish were purchased from China Zebrafish Resource Center (Wuhan, China). Zebrafish were bred in an aquarium system (28 ± 1 °C; 14 hours of light and 10 hours of dark light). These fish were fed with three to four times a day of newly hatched Artemia (Artemia direct sale, USA). All fish experiments were performed according to the regulations of the Animal Experiment Ethics Committee of Northwest Normal University.

[0073] 2. Zebrafish gene editing

[0074] sgRNAs were designed based on zebrafish cbsa and cbsb gene sequences. Sequences of sequencing primers are summarized in Table 1. sgRNAs were synthesized by Genscript (China). Cas9 protein was purchased from Takara (Japan). To generate the founder population (F0), fertilized eggs were collected within 5 minutes after wild type female and male fish naturally laid eggs. Newly fertilized zebrafish embryos were microinjected at the single cell stage. Under a stereomicroscope, sgRNA and Cas9 protein were co-injected into embryos using a glass micropipette connected to a micromanipulator. To compare and evaluate mutagenesis efficiency, embryos without microinjection were designated as wild type (WT) and used as controls. Mutations were detected by PCR genotyping and sequencing. To identify germline transmitted mutations, the remaining F0 founder embryos were raised to adulthood and mated with wild type fish to generate a heterozygous F1 population (+ / -). Adult F1 mutants carrying the same mutant genotype were mated to generate F2 populations, which included some individuals carrying homozygous (- / -) genotype.

[0075] Table 1 Sequencing primer sequences

[0076]

[0077] 3. Homocysteine level determination

[0078] From each group, 30 larvae were collected from wild type (WT) and cbsb mutant zebrafish and transferred into pre-chilled EP tubes containing PBS. Samples were homogenized on ice using a cell disrupter to ensure minimal protein degradation. The homogenate was then centrifuged at 5000 x g for 10 minutes at 4°C to obtain the supernatant for further analysis. The total protein content and homocysteine (Fankew) level in the supernatant were quantified using commercially available spectrophotometric diagnostic kits following the manufacturer’s instructions. Each sample was analyzed in triplicate to ensure the repeatability and accuracy of the measurements. The results are shown in Figure 1 As shown, by drug treatment of cbsb deleted zebrafish, the homocysteine level of AG-270 and IDE-397 treatment groups showed a decreasing trend compared to the cbsb mutant zebrafish group, in which the AG-270 treatment group significantly reduced the homocysteine level, showing that MAT2A inhibitors can effectively reduce the content of homocysteine.

[0079] 4. Drug treatment of zebrafish embryos

[0080] Mutant zebrafish embryos at 6-somite stage (post-fertilization) were chosen for drug screening. The chorion of each embryo was carefully removed under a stereomicroscope using fine forceps. The denuded embryos were then transferred into 24-well plates with 10 embryos per well and 6 replicates per experimental group. Wild-type and cbsb mutant zebrafish larvae were raised in distilled water as control groups. For experimental groups, different concentrations of test drugs (AG-270, IDE-397) were added to the wells. The drug solutions were refreshed daily to ensure continuous exposure. On the third day post-fertilization, the lenses of zebrafish larvae were examined using a Leica microscope (Leica, Germany) and imaged. The lens diameter was quantified using ImageJ software to assess morphological changes. Additionally, zebrafish larvae from each concentration group were collected in EP tubes for biochemical analysis. The homocysteine content in larvae was measured using a commercially available homocysteine detection kit (Fankew) following the manufacturer's instructions.

[0081] Results, as shown in Figure 2 , the cbsb mutant zebrafish eyes, lenses significantly reduced during development, by drug treatment of cbsb mutant zebrafish, as shown in Figure 3 , it was found that AG-270, IDE-397 can alleviate the phenotype of lens reduction.

[0082] 5. O-dianisidine staining method to determine hemoglobin activity

[0083] The hemoglobin (Hb) activity of zebrafish larvae was evaluated using o-dianisidine staining method at 3 days post-fertilization (dpf). Thirty larvae were collected and transferred into 1.5 mL EP tubes. Freshly prepared o-dianisidine staining solution (600 μL per tube) was added, consisting of 40% ethanol, 0.01 M sodium acetate, 0.65% H2O2, and 0.6 mg / mL o-dianisidine. The staining reaction was carried out at room temperature for 15 minutes and protected from light to prevent photodegradation. After staining, the larvae were rinsed three times with PBST (phosphate-buffered saline with Tween 20) to remove excess dye. The o-dianisidine staining method relies on the catalytic activity of Hb, which mediates the H2O2-driven oxidation of o-dianisidine, resulting in deep red coloration of Hb-positive cells (19-21). After staining, the tail vein and heart thrombus regions of the larvae were examined and photographed using a Leica microscope (Leica, Germany). The staining intensity of red blood cells was quantified using ImageJ software to assess Hb activity.

[0084] Results, as shown in Figure 4 , the cbsb mutant zebrafish tail vein embolism, heart back blood reduction, by drug treatment of cbsb mutant zebrafish, it was found that AG-270, IDE-397 can alleviate the phenotype of heart back blood reduction Figure 5 .

[0085] 6. Alizarin red staining

[0086] After fertilization, wild type (WT) and cbs mutant zebrafish embryos were incubated to 8 days post fertilization (dpf), respectively. Then the larvae were fixed in 4% paraformaldehyde for 1 hour at room temperature. To remove pigmentation, the fixed larvae were bleached in a solution containing 2% KOH and 3% H2O2. Subsequently, the samples were stained with alizarin red solution (0.05% alizarin red, 1% KOH and 2% Triton X-100) overnight to observe the mineralization in the skull. After staining, the larvae were washed with a solution of 1% KOH and 2% Triton X-100 to remove excess dye. Then the stained specimens were imaged using a stereomicroscope to capture detailed morphological features of the skeletal structure.

[0087] The results are shown in Figure 6 As shown in Table 1, the cbs mutant zebrafish has a weakened calcium absorption capacity and a tendency of osteoporosis. After drug treatment of the cbs mutant zebrafish, it is found that AG-270 and IDE-397 can alleviate the phenotype of weakened calcium absorption capacity. Figure 7

[0088] Example 2 - Therapeutic effect of MAT2A inhibitor on experimental model of homocysteinemia in mice

[0089] 1. Preparation of experimental model of homocysteinemia in mice

[0090] Hybrid CBS-deficient mice (CBS+ / -) in C57BL / 6 background were purchased from Charles River and then bred with C57BL / 6 mice. CBS+ / - mice were bred to produce homozygous CBS- / - mice. Tail biopsy was performed at 1 week of age, and polymerase chain reaction was performed to genotype the targeted Cbs allele. Wild type (CBS+ / -), hybrid and homozygous mice were used at 2-3 weeks of age. Age-matched mice were compared in each experiment. All mouse experiments were performed in accordance with the regulations of the Animal Experiment Ethics Committee of Northwest Normal University.

[0091] 2. Drug treatment of mouse model

[0092] The control group was 3 WT wild type mice and 3 cbs deletion mice; the experimental group was divided into AG-270 group and IDE-397 group, and the control group was given the same dose of DMSO, and the samples were taken after 3 days of administration.

[0093] 3. Determination of homocysteine and alanine aminotransferase (ALT) activity

[0094] ​To measure homocysteine in mice, blood was collected from the eyeball. The collected blood was kept at room temperature for 2 hours, and centrifuged at 3000 rpm for 10 minutes at 4°C. The supernatant was collected; the liver tissue was taken, lysed using PBS, and centrifuged at 5000 x g for 10 minutes, and the supernatant was taken to measure the Hcy concentration of the mice using a homocysteine detection kit. Liver tissue was isolated from wild-type and mutant group mice. The liver tissue of each group was transferred to a pre-cooled EP tube containing extraction buffer. The tissue was homogenized in an ice bath to ensure minimal protein degradation. The homogenate was then centrifuged at 3500 x g for 10 minutes at 4°C to separate the supernatant, which was collected for further analysis. The total protein content and ALT activity in the supernatant were quantified using commercially available spectrophotometric diagnostic kits according to the manufacturer's protocol. Each sample was analyzed three times to ensure the repeatability and accuracy of the measurements.

[0095] The results, as shown in Figure 8 , by treating the screened homozygous CBS mice, it was found that AG-270, IDE-397 can effectively reduce the level of homocysteine, and can alleviate the increase of liver inflammation index ALT. And its survival rate has also improved.

[0096] Example 3 - Therapeutic effect of MAT2A inhibitors on human liver and blood homocysteinemia

[0097] 1. Experimental method

[0098] Liver tissue was taken from patients who underwent surgery for cancer at the Second Hospital of Lanzhou University, and after 2h induction with Met, the homocysteine content was detected. After 2h treatment with AG270 and IDE397, the homocysteine content after treatment was further detected. Two CBS mutant patients were found at the Second Hospital of Lanzhou University, and the blood of the two CBS mutant patients was incubated with AG270 and IDE397 for 2h, and the homocysteine content after incubation was detected. Before using the donated liver and blood samples, written informed consent was obtained from each patient. This study has been approved by the Institutional Review Board of the Second Hospital of Lanzhou University.

[0099] 2. Determination of therapeutic effect

[0100] Each patient was given AG-270, IDE-397 treatment, and the blood sample before drug administration was used as a control, and the results are shown in Figure 9 , using AG-270, IDE-397 drug treatment of CBS deletion patients, the content of homocysteine has a significant downward trend, among which IDE-397 drug treatment significantly reduces the content of homocysteine.

[0101] As Figure 10As shown, after AG-270 and IDE-397 treatment, the contents of methionine and homocysteine were significantly reduced compared with the liver homocysteine content before drug administration.

[0102] Example 4 - Drug treatment of methionine diet-induced high homocysteine zebrafish model

[0103] Drug treatment was performed on the zebrafish model with elevated homocysteine levels induced by high methionine diet. After 8 days of treatment with AG-270 and IDE-397, the results were as follows Figure 11 As shown, AG-270 and IDE-397 can reduce the contents of homocysteine and methionine, and the effect of AG-270 on reducing homocysteine and methionine is more significant.

[0104] As can be seen from the above, by using MAT2A inhibitors, homocysteine regeneration can be blocked, and the content of homocysteine can be effectively reduced. Methionine, as an essential amino acid, can be metabolized by other pathways, and methionine adenosyltransferase inhibitors have good therapeutic effect as a new type of high homocysteine treatment drug, significantly alleviate liver inflammation indicators, and have good therapeutic effect for high homocysteine patients. The present application believes that MAT2A can be used as an effective target for high homocysteine treatment, and provides a new possibility for the treatment of high homocysteine.

[0105] The above has described the basic concepts, and it is obvious that the above detailed disclosure is only as an example for those skilled in the art, and does not constitute a limitation on the present specification. Although it is not explicitly stated here, those skilled in the art can make various modifications, improvements and corrections to the present specification. Such modifications, improvements and corrections are suggested in the present specification, so such modifications, improvements and corrections still belong to the spirit and scope of the exemplary embodiments of the present specification.

[0106] At the same time, specific words are used in the present specification to describe the embodiments of the present specification. As "one embodiment", "an embodiment", and / or "some embodiments" means a certain feature, structure or characteristic related to at least one embodiment of the present specification. Therefore, it should be emphasized and noted that the "an embodiment" or "one embodiment" or "one alternative embodiment" mentioned in different places in the present specification does not necessarily refer to the same embodiment. In addition, some features, structures or characteristics in one or more embodiments of the present specification can be properly combined.

[0107] Finally, it should be understood that the embodiments described herein are only given by way of example and that other modifications can occur to persons skilled in the art. Therefore, the scope of the present description is not intended to be limited to the embodiments described herein but is only limited by the claims that follow.

Claims

1. Use of a MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof in the manufacture of a medicament for treating hyperhomocysteinemia or its complications.

2. Use according to claim 1, wherein The MAT2A inhibitor is a substance that reduces the expression of MAT2A and / or hinders the function of MAT2A.

3. The use according to claim 1, wherein The MAT2A inhibitor is selected from AG-270 or IDE-397.

4. Use according to claim 1, characterized in that, The complications are selected from any one or more of ocular complications, cardiovascular diseases, skeletal system diseases, liver damage, central nervous system diseases, developmental retardation.

5. The use according to claim 1, wherein the compound is ###0002### The MAT2A inhibitor has at least one of the following effects: (1) alleviating liver inflammation; (2) alleviating lens shrinkage; (3) improving heart function; (4) improving vascular wall thickening; (5) improving calcium absorption capacity.

6. The use according to claim 1, wherein The MAT2A inhibitor is used alone or in combination with other drugs.

7. A composition for treating hyperhomocysteinemia, the composition comprising a MAT2A inhibitor or a pharmaceutically acceptable salt, ester, isomer, prodrug, polymorph or solvate thereof as claimed in any one of claims 1 to 5, and a pharmaceutically acceptable excipient.

8. The composition of claim 7, wherein The pharmaceutically acceptable excipient is selected from one or more of stabilizers, excipients, antioxidants, buffers, preservatives, surfactants, chelating agents, binders.

9. The composition of claim 7, wherein The composition comprises one or more of injections, sterile powders for injection, tablets, pills, capsules, lozenges, elixirs, powders, granules, syrups, solutions, tinctures, aerosols, powder sprays, or suppositories.

10. Use of the composition as claimed in any one of claims 7 to 9 in the manufacture of a product for treating hyperhomocysteinemia or its complications.

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