Use of natural derivative in preparation of drug for treating cardiomyopathy and / or myocardial fibrosis

Abstract

The present invention relates to the technical field of biomedicine and discloses use of a hypoxanthine derivative in the preparation of a drug for treating cardiomyopathy and / or myocardial fibrosis. By assaying pharmacological activities of structurally modified and altered hypoxanthine derivatives in treating cardiomyopathy and / or myocardial fibrosis, the pharmacological activities of such compounds were tested in various animal disease models, and data on activities for preventing and treating different types of cardiomyopathy and / or myocardial fibrosis were provided. It is proven that the hypoxanthine derivatives all have good activities, the effects are significantly superior to those of a clinically commonly used drug, nintedanib, and the effects are also significantly superior to those of hypoxanthine derivatives A, B, and C and other hypoxanthine derivatives in the prior art. The hypoxanthine derivative of the present invention can provide a novel skeleton for the screening of new compounds for the preparation of a drug for treating cardiomyopathy and / or myocardial fibrosis.

Description

Application of a natural derivative in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the use of a hypoxanthine derivative in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis. Background Technology

[0002] Cardiomyopathy is a group of diseases affecting the myocardium (heart muscle), characterized by abnormalities in cardiac structure and function, typically impacting the heart's pumping ability. Cardiovascular disease is currently the leading cause of chronic diseases worldwide, and most cardiovascular diseases, including cardiomyopathy, hypertension, coronary artery disease, atrial fibrillation, and arteriosclerosis, undergo a common pathological stage—myocardial fibrosis. Myocardial fibrosis is primarily characterized by extracellular matrix remodeling and excessive collagen accumulation, which can promote sudden cardiac death, ventricular tachycardia, left ventricular dysfunction, and heart failure, and is a prognostic factor for adverse outcomes in various cardiac pathologies. Although initially myocardial fibrosis is a compensatory mechanism to maintain the functional and structural integrity of a damaged heart, if pathological factors persist, it can lead to severely adverse prognoses and heart failure.

[0003] Heart failure (HF) includes idiopathic heart failure and secondary heart failure. It is the end stage of various cardiovascular diseases. Myocardial fibrosis is an inevitable process in the progression of heart diseases such as hypertension, coronary heart disease, and cardiomyopathy to heart failure. It is an extremely important pathophysiological process of heart failure. At present, there is no treatment that has a significant effect on heart failure and myocardial fibrosis, which leads to the continuous development of cardiomyopathy and a still high mortality rate.

[0004] Currently, only two anti-fibrotic drugs are used clinically: nintedanib and pirfenidone. Because the pathogenesis of fibrosis differs across organs, nintedanib and pirfenidone are not very effective against cardiomyopathy and / or myocardial fibrosis. The FDA has only approved their clinical indication for pulmonary fibrosis. Unlike myocardial fibrosis, pulmonary fibrosis is a diffuse fibrotic disease of unknown cause. It is mainly characterized by multiple pathways, including activated inflammatory responses, oxidative stress, and epithelial-mesenchymal transition, after the body is stimulated, leading to the destruction of epithelial cells and further aggravation of the production of fibrotic mediators and extracellular matrix, gradually leading to pulmonary fibrosis.

[0005] Myocardial fibrosis is a crucial intervention target for the clinical prevention and treatment of cardiomyopathy. However, currently, there is a lack of effective treatments to inhibit or reverse it. Existing treatment strategies mainly involve myocardial fibrosis signaling pathways such as the renin-angiotensin-aldosterone system and the TGFβ signaling pathway, but their efficacy is extremely limited. Currently, there are no therapeutic drugs for myocardial fibrosis, and there is an urgent need to develop effective drugs to inhibit or delay its progression, improve heart failure, and increase cardiac function. This is vital for the prevention and treatment of cardiomyopathy and / or myocardial fibrosis.

[0006] Hypoxanthine, also known as 6-hydroxypurine, is a naturally occurring purine compound and a precursor for the synthesis of purine nucleotides in nucleic acids. Currently, there are no reports of hypoxanthine derivatives blocking the progression of cardiomyopathy and / or myocardial fibrosis or reversing pathological damage. We have developed a series of hypoxanthine derivatives that can effectively improve cardiomyopathy and / or myocardial fibrosis. Summary of the Invention

[0007] The purpose of this invention is to propose the application of a hypoxanthine derivative in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, thereby accelerating the development of new drugs for treating cardiomyopathy and / or myocardial fibrosis. The numerous technical effects of the preferred embodiment of this invention are detailed below.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] The first aspect of this invention relates to the use of hypoxanthin derivatives in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, wherein the hypoxanthin derivatives have activity for treating cardiomyopathy and / or myocardial fibrosis, and wherein the hypoxanthin derivatives have one of the following structures:

[0010] in:

[0011] R1 can be any of O, N, C, S or = O;

[0012] R2, R3, and R4 can be chosen as H and C1-C, respectively. 18 Alkyl or halogen-substituted C1-C 18 Alkyl, trifluoromethyl, sulfonyl, sulfonamide, sulfinyl, amino acid, 2-[bis(neopentyloxy)methoxy]phosphonomethoxyethyl, C1-C 18 Fatty acid group, C3-C 12 Heterocyclic groups, C1-C 18 Fatty acids; or C1-C atoms in which R2, R3, and R4 are optionally substituted with oxygen, sulfur, or nitrogen atoms. 18 The alkyl or fatty acid group; when R3 or R4 is substituted, the double bond is attached to the unsubstituted N position, and when all are substituted, there is no double bond.

[0013] The second aspect of this invention relates to compounds having the following structure:

[0014] in:

[0015] R1 can be any of O, N, C, S or = O;

[0016] R2, R3, and R4 can be chosen as H and C1-C, respectively. 18 Alkyl or halogen-substituted C1-C 18 Alkyl, trifluoromethyl, sulfonyl, sulfonamide, sulfinyl, amino acid, 2-[bis(neopentyloxy)methoxy]phosphonomethoxyethyl, C1-C 18 Fatty acid group, C3-C 12 Heterocyclic groups, C1-C 18 Fatty acids; or C1-C atoms in which R2, R3, and R4 are optionally substituted with oxygen, sulfur, or nitrogen atoms. 18 The alkyl or fatty acid group; when R3 or R4 is substituted, the double bond is attached to the unsubstituted N position, and when all are substituted, there is no double bond.

[0017] According to a preferred embodiment, the compound is selected from the group consisting of:

[0018] A third aspect of the present invention relates to pharmaceutical compositions comprising the compounds of the present invention or pharmaceutically acceptable salts thereof.

[0019] According to a preferred embodiment, the pharmaceutical composition further comprises pharmaceutically acceptable excipients or auxiliary ingredients.

[0020] According to a preferred embodiment, the pharmaceutical composition is an oral formulation, an injectable formulation, or a nasal mucosal administration formulation.

[0021] The fourth aspect of the present invention relates to a method for treating cardiomyopathy and / or myocardial fibrosis, by administering an effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention, to an individual in need.

[0022] According to a preferred embodiment, the medication for treating cardiomyopathy and / or myocardial fibrosis includes medications that have the efficacy of preventing and treating cardiomyopathy and / or myocardial fibrosis and their complications.

[0023] According to a preferred embodiment, the drug for treating cardiomyopathy and / or myocardial fibrosis is a formulation prepared by adding pharmaceutically acceptable excipients or auxiliary ingredients, with hypoxanthine derivatives or their salts as the active ingredient.

[0024] According to a preferred embodiment, the formulation is an oral formulation, an injectable formulation, or a nasal mucosal administration formulation.

[0025] According to a preferred embodiment, the myocardial fibrosis includes one or more of idiopathic myocardial fibrosis and secondary myocardial fibrosis.

[0026] Terminology definition:

[0027] The compounds and derivatives provided by this invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature systems.

[0028] The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group. Examples of C1 to C3 alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), and isopropyl (C3).

[0029] The term "pharmaceutically acceptable" means that a carrier, delivery substance, diluent, excipient, and / or the salt formed therefrom is generally chemically or physically compatible with other components constituting a drug dosage form and physiologically compatible with receptors.

[0030] The term "pharmaceutically acceptable salt" refers to acidic and / or basic salts formed by the compounds of this invention with inorganic and / or organic acids and bases, including zwitterionic salts (internal salts) and quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final separation and purification of the compounds. Alternatively, they can be obtained by mixing the aforementioned compounds with appropriate (e.g., equimolar) amounts of an acid or base. These salts may form precipitates in solution and be collected by filtration, or be recovered after solvent evaporation, or be prepared by freeze-drying after reaction in an aqueous medium. The salts described in this invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluoric acids, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates, or trifluoroacetates of the compounds.

[0031] There are no particular limitations on the administration of the compounds or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0032] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with: (a) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin wax; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0033] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0034] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0035] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0036] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0037] 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.

[0038] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers, such as nasal mucosal delivery formulations. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0039] The pharmaceutically acceptable excipients described in this invention refer to substances other than the active ingredient contained in the dosage form.

[0040] The pharmaceutically acceptable adjuvant component described in this invention possesses certain physiological activities. However, the addition of this component does not alter the dominant role of the aforementioned pharmaceutical composition in the disease treatment process; rather, it merely exerts an adjuvant effect. These adjuvant effects are simply the utilization of the known activity of the component, and are a commonly used adjuvant therapy method in the pharmaceutical field. If the aforementioned adjuvant component is used in combination with the pharmaceutical composition of this invention, it should still fall within the scope of protection of this invention.

[0041] Specifically, the hypoxanthin derivatives provided by this invention, when used in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, can significantly improve cardiomyopathy and / or myocardial fibrosis. More importantly, the hypoxanthin derivatives provided by this invention can improve one or more types of idiopathic and secondary cardiomyopathy and / or myocardial fibrosis.

[0042] The hypoxanthin derivatives provided by this invention have at least the following beneficial technical effects:

[0043] This invention relates to the application of hypoxanthine derivatives in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis. The pharmacological activities of these modified hypoxanthine derivatives in treating cardiomyopathy and / or myocardial fibrosis were tested in various animal disease models, including the classic aortic coarctation model and the doxorubicin-induced myocardial injury model. Data on their activity in preventing and treating different types of cardiomyopathy and / or myocardial fibrosis are provided, confirming their good activity and significantly superior efficacy compared to the clinically commonly used drug nintedanib. Their efficacy is also significantly superior to hypoxanthine derivatives A, B, and C, and other hypoxanthine derivatives in the prior art. This invention provides a novel framework for screening new compounds for the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, laying a theoretical foundation for the development of novel lead compounds.

[0044] Brief description of the attached figures

[0045] Figure 1. HE images of the hearts of each experimental group in the in vivo mouse model of heart failure and myocardial fibrosis.

[0046] Figure 2. Masson plots of the hearts in each experimental group in mouse models of heart failure and myocardial fibrosis in vivo. Detailed Implementation

[0047] To make the objectives, advantages, and technical solutions of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other implementations obtained by those skilled in the art without creative effort, including extended studies on the treatment of cardiomyopathy and / or myocardial fibrosis using hypoxanthine derivatives of this invention, are within the scope of protection of this invention.

[0048] The aortic coarctation model and the doxorubicin-induced myocardial injury model are classic models for studying cardiomyopathy, myocardial fibrosis, and heart failure. This application utilizes these two classic models to systematically evaluate the anti-cardiomyopathy, myocardial fibrosis, and heart failure activities of a designed and synthesized hypoxanthine derivative. Through the study of natural product medicinal chemistry and chemical research, many plant endogenous compounds and derivatives have been developed. Modification and alteration of the structure of natural products have yielded many derivatives with excellent pharmacological and chemical activities. The application of the hypoxanthine derivatives provided in this invention in the preparation of drugs for anti-cardiomyopathy and / or myocardial fibrosis demonstrates the effectiveness of these compounds in various animal disease models. Data on their activity in preventing and treating different types of cardiomyopathy and / or myocardial fibrosis are provided, confirming their good activity and significantly superior effects compared to the clinically commonly used positive control drug nintedanib, as well as hypoxanthine derivatives A, B, and C, and other hypoxanthine derivatives in the prior art. This invention provides a novel framework for screening new compounds for the preparation of drugs against cardiomyopathy and / or myocardial fibrosis, and lays a theoretical foundation for the development of novel lead compounds.

[0049] The structures of hypoxanthine derivatives A (control 1), B (control 2), and C (control 3) are shown below:

[0050] Hypoxanthine derivative A (control 1) was commercially available. Compounds B (control 2) and C (control 3) were prepared according to the method used to prepare compounds 3 and 4 as described below.

[0051] The application of the hypoxanthine derivatives provided by the present invention in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis is described in detail below with reference to Examples 1 and 2.

[0052] Example 1: Preparation method of compounds 1 to 18

[0053] According to a preferred embodiment, compounds 1 to 18 are prepared by alkylation of hypoxanthin.

[0054] This embodiment provides preparation methods for 18 compounds, and the structures of all obtained compounds were determined by nuclear magnetic resonance spectroscopy and mass spectrometry.

[0055] Preparation of Compound 1 and Compound 5

[0056] The synthetic routes for compounds 1 and 5 are shown below:

[0057] Reaction: (1) At 0℃, 3.45g NaH (4.5eq) was added to a 250mL four-necked flask. After evacuating the gas 3 times, 40mL (8V) of anhydrous THF was slowly added under N2 atmosphere; (2) 12.3mL of isopropanol (4.5eq) was slowly added dropwise to the system and the reaction was allowed to proceed for 30min; (3) Subsequently, 5g of compound 1 (1.0eq) was slowly added dropwise to a 100mL (20V) isopropanol mixture to the reaction system; (4) The temperature was raised to 80℃ and the system was allowed to react for 10h.

[0058] Post-processing: (1) After the reaction was complete, water was added to quench the reaction and acetic acid was added to neutralize the pH to 8-10; (2) Ethyl acetate was extracted 5 times and the organic phases were combined; (3) The organic phase was dried with anhydrous sodium sulfate, concentrated and then separated by rapid silica gel column chromatography to obtain a pale yellow solid; (4) TLC monitoring: developing solvent: dichloromethane / methanol = 10:1 Rf(compound 1) = 0.4.

[0059] The relevant spectral data for compounds 1 and 5 are as follows:

[0060] Compound 1: ¹H NMR (400MHz, DMSO-d6) δ 13.33 (s, ¹H), 8.44 (s, ¹H), 8.32 (s, ¹H), 5.55 (hept, J = 6.1Hz, ¹H), 1.37 (d, J = 6.2Hz, 6H). HRMS (ESI-TOF) calculated for C8H10N4OH+[M+H+]: 179.09; found 179.10.

[0061] Compound 5: 11H NMR (400MHz, DMSO-d6) δ 13.37 (s, 1H), 8.47 (s, 1H), 8.35 (s, 1H), 5.57 (hept, J = 6.2Hz, 1H), 1.39 (d, J = 6.2Hz, 6H). HRMS (ESI-TOF) [M+H+]: 179.09; found 179.00.

[0062] Preparation of compounds 2, 4 and 6

[0063] The synthetic routes for compounds 2, 4, and 6 are shown below:

[0064] The relevant spectral data for compounds 2, 4, and 6 are as follows:

[0065] Compound 2: 1 found 179.30.

[0066] Compound 4: 1 H NMR(400MHz,Chloroform-d)δ8.58(s,1H),8.07(s,1H),5.63(hept,J=6.2Hz,1H),4.90(hept,J =6.7Hz,1H),1.59(d,J=6.8Hz,6H),1.44(d,J=6.2Hz,6H).HRMS(ESI-TOF)[M+H+]:221.27; found 221.20.

[0067] Compound 6: 1 found 179.30.

[0068] Preparation of compound 3

[0069] The synthetic route for compound 3 is shown below:

[0070] The relevant spectral data for compound 3 are as follows:

[0071] Compound 3: 1 H NMR (400MHz, DMSO-d6) δ8.43 (s, 1H), 8.19 (s, 1H), 5.08 (hept, J = 6.9 Hz, 1H), 4.71 (hept, J = 6. 8Hz,1H),1.52(d,J=6.8Hz,6H),1.41(d,J=6.9Hz,6H).HRMS(ESI-TOF)[M+H+]:221.27; found 221.30.

[0072] Preparation of compounds 7 to 18

[0073] Compounds 7 through 18 were prepared using the same method as compound 3 described above.

[0074] The relevant spectral data for compounds 7 through 18 are as follows:

[0075] Compound 7: 1 found 193.10.

[0076] Compound 8: 1 H NMR(500MHz,Chloroform-d)δ8.55(s,1H),7.95(d,J=0.9Hz,1H),4.85(heptd,J=5.2,0.5Hz,1H) ,4.62(q,J=6.5Hz,2H),1.57(s,3H),1.50(t,J=6.5Hz,3H).HRMS(ESI-TOF)[M+H+]:207.12; found 207.10.

[0077] Compound 9: 1H NMR (500MHz, Chloroform-d) δ8.51 (s, 1H), 7.57 (d, J = 0.3Hz, 1H), 4.57 (heptd, J = 5.0, 0.7Hz, 1H), 4.61 (t, J = 5. 4Hz,2H),1.90(qt,J=7.3,5.9Hz,2H),1.63(s,6H),1.15(t,J=7.4Hz,3H).HRMS(ESI-TOF)[M+H+]:221.13; found 221.10.

[0078] Compound 10: 1 H NMR(500MHz,Chloroform-d)δ8.37(s,1H),5.33–5.27(m,3H),4.74–4.64(m,2H),1.25 (d,J=6.2Hz,6H),1.17(dd,J=6.7,5.2Hz,12H).HRMS(ESI-TOF)[M+H+]:265.20; found 265.20.

[0079] Compound 11: 1 H NMR (500MHz, Chloroform-d) δ7.99–7.88 (m, 2H), 4.83 (pd, J=5.6, 0.8Hz, 1H), 2.32 (s, 3H), 1.53 (s, 6H). HRMS (ESI-TOF) [M+H+]: 221.10; found 221.10.

[0080] Compound 12: 1 H NMR(500MHz,Chloroform-d)δ7.99–7.90(m,2H),4.85(heptd,J=5.2,0.5Hz,1H),2.54( q,J=7.7Hz,2H),1.55(s,3H),1.19–1.11(m,3H).HRMS(ESI-TOF)[M+H+]:235.10; found 235.10.

[0081] Compound 13: 1 H NMR(500MHz,Chloroform-d)δ8.05(s,1H),7.87(s,1H),4.35(s,1H),3.92(s,3H),1.37(s,6H).HRMS(ESI-TOF)[M+H+]:193.11; found193.11.

[0082] Compound 14: 1H NMR(500MHz,Chloroform-d)δ7.98(s,1H),7.87(s,1H),4.37(s,1H),4.29(d,J=12.2Hz,1H ),4.23(d,J=12.3Hz,1H),1.47(s,3H),1.43(s,6H).HRMS(ESI-TOF)[M+H+]:207.12; found 207.12.

[0083] Compound 15: 1 H NMR(500MHz,Chloroform-d)δ7.87(s,1H),7.83(s,1H),4.73(s,1H),4.39(s,1H),1.57(s,6H),1.42(s,6H).HRMS(ESI-TOF)[M+H+]:221.14; found 221.14.

[0084] Compound 16: 1 H NMR(500MHz,Chloroform-d)δ8.32(s,1H),7.87(s,1H),4.72(s,1H),4.13(s,3H),1.33(s,6H).HRMS(ESI-TOF)[M+H+]:193.11; found193.11.

[0085] Compound 17: 1 H NMR(500MHz,Chloroform-d)δ8.32(s,1H),7.92(s,1H),4.72(s,1H),4.53(s,2H),1.64(s,3H),1.34(s,6H).HRMS(ESI-TOF)[M+H+]:207.12; found 207.12.

[0086] Compound 18: 1 H NMR(500MHz,Chloroform-d)δ8.32(s,1H),7.87(s,1H),4.77(s,1H),4.65(s,1H),1.62(s,6H),1.33(s,6H).HRMS(ESI-TOF)[M+H+]:221.28; found 221.28.

[0087] Example 2: The activity of compounds 1 to 18 obtained in Example 1 against aortic stenosis-induced cardiomyopathy, heart failure and myocardial fibrosis.

[0088] Experimental Methods: An in vivo model of cardiomyopathy and myocardial fibrosis was constructed. The coronary artery ligation model can fully simulate the acute and compensatory phases of myocardial fibrosis after myocardial infarction, and is a typical model for studying cardiomyopathy, heart failure, and myocardial fibrosis.

[0089] SPF-grade C57BL / 6 mice (weighing approximately 22–25 g) were randomly and evenly divided into several groups, including a blank control group, a model group, a compound 1-18 intervention model group, and a control group 1-4 drug intervention model group (compound AC intervention and control group 4 nintedanib), with 15 mice in each group. They were housed in an SPF-grade animal center for 7 days. The mice were fasted for 12 hours the day before the experiment, anesthetized with isoflurane, and maintained on a ventilator. The thorax of the mice was opened, and the left anterior descending coronary artery was ligated. After ligation, the thoracic cavity was sutured. The blank control group underwent only thorax opening without ligation of the blood vessels. Three days after model establishment, mice in each group were given intraperitoneal injections of the drug. The blank control group and the model group were given the same volume of physiological saline as the drug group. The model group and the drug intervention group were given an appropriate volume of 60 mg / kg / day of drug compound 1-18, control group 1-3 (compounds A, B, and C, respectively), and control group 4 nintedanib, twice a day for 17 days until the model was established 21 days later. Then, the cardiac function of the mice was detected by ultrasound. The mice were anesthetized with 0.8% sodium pentobarbital solution (10 mL / kg), the heart was removed and weighed, the left ventricle was homogenized and the myocardial enzyme LDH content was measured to evaluate the cardiac function of the mice. The paraffin sections of the heart tissue were stained with HE and Masson staining to evaluate the severity of myocardial fibrosis and to calculate the area of ​​myocardial fibrosis.

[0090] The cardiac HE results (Figure 1) and cardiac HE pathological scores (Table 1) of each group showed that, compared with the control group, the model group had enlarged and degenerated cells, most cells had no obvious gaps, and myocardial fibers were broken and fused in a wavy pattern. Compared with the model group, the cardiac tissue of the compound 1-18 intervention group had reduced myocardial cell swelling and reduced myocardial fiber breakage. It was also observed that the cardiac inflammatory cell infiltration was significantly reduced in the compound 1-18 drug intervention group, indicating that the hypoxanthine derivative 1-18 has a significant effect on anti-heart failure and myocardial fibrosis, and is superior to the control group 1-3 (hypoxanthine analog AC) and the control group 4 nintedanib.

[0091] Table 1. Cardiac HE pathology score

[0092] The results of Masson staining (Figure 2) and Masson staining collagen volume fraction (Table 2) showed that, compared with the control group, the model group had hypertrophic cardiomyocytes, irregular cell morphology, disordered intercellular arrangement, and a large amount of purple collagen fibers aggregated between tissues. In the intervention group of compounds 1-18, the cardiac tissue showed slightly more regular cardiomyocyte morphology and slightly disordered intercellular arrangement compared with the model group, and a significant reduction in purple collagen fibers (P < 0.05), with significantly better activity than the control group (nintedanib).

[0093] Table 2. Semi-quantitative analysis results of Masson staining collagen volume fraction in heart tissue of mice in each group.

[0094] Table 3 shows that compared with the control group, the ejection fraction (EF) and fractional shortening (FS) of mice in the model group were significantly decreased (P < 0.05), while the decrease in EF and FS in the compound 1-18 intervention group was lower than that in the model group, and also lower than that in control groups 1-3 and 4 (nintedanib group). Compared with the control group, the ventricular end-diastolic diameter (LVIDd) of mice in the model group was significantly increased (P < 0.05), while the increase in LVIDd in the compound 1-18 intervention group was lower than that in the model group, and also lower than that in control groups 1-3 and 4 (nintedanib group). These results indicate that compound 1-18 intervention can significantly improve cardiac function associated with heart failure and myocardial fibrosis.

[0095] Table 3. Results of Ultrasound Examination of Cardiac Function

[0096] As shown in Table 4, compared with the control group, the model group showed a significant increase in cardiac index and lactate dehydrogenase levels. The increases in cardiac index and lactate dehydrogenase levels in compounds 1-18 were lower than those in the model group, and also lower than those in control groups 1-3 and 4 (nintedanib group). These results indicate that intervention with compounds 1-18 can significantly improve cardiac damage caused by heart failure and myocardial fibrosis.

[0097] Table 4. Cardiac Index and Myocardial Enzyme Content in the Heart

[0098] Example 3: The activity of compounds 1 to 18 obtained in Example 1 against doxorubicin-induced cardiomyopathy, heart failure and myocardial fibrosis.

[0099] Experimental methods: An in vivo model of doxorubicin-induced cardiomyopathy and myocardial fibrosis was established. Male SPF-grade C57BL / 6 mice (22g±3g) were injected with DOX (5mg / kg) via the tail vein once a week for 4 weeks, with a cumulative dose of 20mg / kg. Twenty-eight days after the first injection of doxorubicin, mice in each group were administered medication by gavage. The blank control group and the model group were given the same volume of physiological saline as the drug group. The model group and the drug intervention group were given an appropriate volume of 60 mg / kg / day of drug compound 1-18, control group 1-3 (compounds A, B, and C, respectively), and control group 4 nintedanib, twice a day for 14 days until the model was established. Then, the cardiac function of the mice was detected by ultrasound. The mice were anesthetized with 0.8% sodium pentobarbital solution (10 mL / kg), the heart was removed and weighed, the left ventricle was homogenized and the myocardial enzyme LDH content was measured to evaluate the cardiac function of the mice. The paraffin sections of the heart tissue were stained with HE and Masson staining to evaluate the severity of myocardial fibrosis and to calculate the area of ​​myocardial fibrosis.

[0100] The results of Masson staining collagen volume fractions in each group (Table 5) show that the collagen volume fraction in the model group was significantly increased compared with that in the control group, while the collagen volume fraction in the intervention group of compound 1-18 was significantly decreased (P<0.05), and its activity was significantly better than that of control group 1-3 and control group 4 (nintedanib).

[0101] Table 5. Semi-quantitative analysis results of Masson staining collagen volume fraction in mouse heart tissue of each group.

[0102] Table 6 shows that, compared with the control group, the ejection fraction (EF) and fractional shortening (FS) of the model group mice were significantly reduced (P < 0.05). However, in the mice treated with compounds 1-18, the decrease in EF and FS was smaller than that in the model group, and also lower than that in control groups 1-3 and 4 (i.e., the nintedanib group). On the other hand, compared with the control group, the ventricular end-diastolic diameter (LVIDd) of the model group mice was significantly increased (P < 0.05). However, the increase in LVIDd in the compound 1-18 intervention groups was smaller than that in the model group, and also lower than that in control groups 1-3 and 4 (nintedanib group). These findings suggest that intervention with compounds 1-18 can significantly improve the doxorubicin-induced cardiomyopathy model, heart failure, and cardiac function indicators related to myocardial fibrosis.

[0103] Table 6. Results of Ultrasound Examination of Cardiac Function

[0104] As shown in Table 4, compared with the control group, the model group showed a significant increase in cardiac index and lactate dehydrogenase levels. The increases in cardiac index and lactate dehydrogenase levels in compounds 1-18 were lower than those in the model group, and also lower than those in control groups 1-3 and 4 (nintedanib group). These results indicate that intervention with compounds 1-18 can significantly improve cardiac damage caused by heart failure and myocardial fibrosis.

[0105] Table 7. Cardiac Index and Myocardial Enzyme Content in the Heart

[0106] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

[0107] References

[0108] 1. Li Xue, Zhao Yajing, Zhang Qin, Zheng Shaoying, Zhou Mingshuang, Zhang Wen. Research progress on the regulatory mechanism of myocardial fibrosis [J]. China Medicine, 2024, 19(02):285-288.

[0109] 2. Huang Bin, Zheng Jinxu, Zhang Jun. Non-coding RNA and stem cell abnormalities in the progression of idiopathic pulmonary fibrosis [J]. Chinese Journal of Tissue Engineering Research, 2023, 27(1):130-137.

[0110] 3. Zeng Min, Wang Hua. Current status and prospects of diagnosis and treatment of myocardial fibrosis [J]. Chinese Journal of Medical Frontiers (Electronic Edition), 2023, 15(03):64-71.

[0111] 4. Zhou Zimo, Zhou Chi, Yang Yayuan, et al. Research progress on the pathogenesis and treatment of myocardial fibrosis [J]. Journal of Hubei University of Science and Technology (Medical Edition), 2022, 36(05):452-456. DOI:10.16751 / j.cnki.2095-4646.2022.05.0452.

Claims

1. The use of a hypoxanthine derivative in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, characterized in that, The hypoxanthin derivative has therapeutic activity for cardiomyopathy and / or myocardial fibrosis, and the hypoxanthin derivative has one of the following structures: in: R1 can be any of O, N, C, S or = O; R2, R3, and R4 can be chosen as H and C1-C, respectively. 18 Alkyl or halogen-substituted C1-C 18 Alkyl, trifluoromethyl, sulfonyl, sulfonamide, sulfinyl, amino acid, 2-[bis(neopentyloxy)methoxy]phosphonomethoxyethyl, C1-C 18 Fatty acid group, C3-C 12 Heterocyclic groups, C1-C 18 Fatty acids; or C1-C atoms in which R2, R3, and R4 are optionally substituted with oxygen, sulfur, or nitrogen atoms. 18 The alkyl or fatty acid group; when R3 or R4 is substituted, the double bond is attached to the unsubstituted N position, and when all are substituted, there is no double bond.

2. The use of the hypoxanthine derivative according to claim 1 in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, characterized in that, The hypoxanthine derivatives mentioned are one or more of the following compounds:

3. The use of the hypoxanthine derivative according to claim 1 in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, characterized in that, The drugs mentioned for treating cardiomyopathy and / or myocardial fibrosis include those that have the efficacy of preventing and treating cardiomyopathy and / or myocardial fibrosis and their complications.

4. The use of the hypoxanthine derivative according to claim 1 in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, characterized in that, The aforementioned drug for treating cardiomyopathy and / or myocardial fibrosis is a preparation made by adding pharmaceutically acceptable excipients or auxiliary ingredients, with hypoxanthine derivatives or their salts as the active ingredient.

5. The use of the hypoxanthine derivative according to claim 4 in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, characterized in that, The preparation is an oral preparation, an injectable preparation, or a nasal mucosal delivery preparation.

6. The use of the hypoxanthine derivative according to claim 1 in the preparation of drugs for treating cardiomyopathy and / or myocardial fibrosis, characterized in that, The myocardial fibrosis includes one or more of idiopathic myocardial fibrosis and secondary myocardial fibrosis.

7. Compounds having the following structure: in: R1 can be any of O, N, C, S or = O; R2, R3, and R4 can be chosen as H and C1-C, respectively. 18 Alkyl or halogen-substituted C1-C 18 Alkyl, trifluoromethyl, sulfonyl, sulfonamide, sulfinyl, amino acid, 2-[bis(neopentyloxy)methoxy]phosphonomethoxyethyl, C1-C 18 Fatty acid group, C3-C 12 Heterocyclic groups, C1-C 18 Fatty acids; or C1-C atoms in which R2, R3, and R4 are optionally substituted with oxygen, sulfur, or nitrogen atoms. 18 The alkyl or fatty acid group; when R3 or R4 is substituted, the double bond is attached to the unsubstituted N position, and when all are substituted, there is no double bond.

8. The compound according to claim 7, selected from the group consisting of:

9. A pharmaceutical composition comprising the compound of claim 7 or 8 or a pharmaceutically acceptable salt thereof.

10. The pharmaceutical composition according to claim 9, further comprising a pharmaceutically acceptable excipient or auxiliary ingredient.

11. The pharmaceutical composition according to claim 9 or 10, wherein it is an oral formulation, an injectable formulation, or a nasal mucosal administration formulation.

12. A method for treating cardiomyopathy and / or myocardial fibrosis, characterized in that, Administer to an individual in need an effective amount of the compound of claim 7 or 8 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claims 9-11.

13. The method according to claim 12, characterized in that, The myocardial fibrosis includes one or more of idiopathic myocardial fibrosis and secondary myocardial fibrosis.

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