Pharmaceutical composition for preventing and / or treating heart system diseases of mammals

Abstract

The embodiment of the invention discloses a medicine composition for preventing and / or treating heart system diseases of mammals. The medicine composition comprises a mitochondrial function optimizing substance and a medicine for treating heart system related diseases. According to the embodiment of the invention, the pharmaceutical composition has the beneficial effects of synergistic interaction or reduction of side effects of drugs in the aspect of preventing and / or treating heart system diseases of mammals.

Description

Technical Field

[0001] This application relates to the field of pharmaceutical compositions, and more particularly to a pharmaceutical composition for the prevention and / or treatment of diseases of the mammalian cardiac system. Background Technology

[0002] The health of companion and ornamental animals is attracting increasing attention. Multiple factors, such as overnutrition and aging, are causing increasingly common cardiac diseases in these animals. Consequently, the treatment of cardiac diseases in animals is becoming a focus of industry attention.

[0003] Currently available medications mainly include: angiotensin-converting enzyme inhibitors (ACEIs), angiotensin receptor antagonists, phosphokinase inhibitors, dihydropyridine calcium channel blockers, platelet aggregation inhibitors, lipid-lowering drugs, beta-blockers, antiarrhythmics, vasodilators, 5-HT antagonists, and diuretics. It is difficult to find a single drug that can effectively and comprehensively treat cardiovascular diseases in companion animals and ornamental animals. Veterinarians often use multiple single-drug combinations, which involves not only the rational combination of drug types but also the appropriate dosage, the pet owner's correct understanding and administration, and even the issue of animal compliance with complex and repeated medication administration.

[0004] Since it is difficult to find single-component drugs specifically for the complex cardiovascular diseases in animals, exploring better drug combinations becomes essential. Addressing the complexities of diagnosing and treating animal cardiovascular diseases, as well as the challenges of medication adherence, finding a solution that effectively addresses both dosage and drug combination is of significant practical importance for improving the entire industry, both from the perspective of treating animal cardiovascular diseases and enhancing drug administration and compliance.

[0005] However, no animal-specific compound formulation currently offers a satisfactory solution to these challenges. For instance, patent application CN102727500A discloses a compound formulation combining pimoxandan and aminophylline, which utilizes aminophylline to alleviate asthma symptoms. However, the addition of aminophylline fails to overcome the risk of excessive cardiac stimulation from pimoxandan. Not all cardiovascular diseases lead to severe asthma; more critical aspects such as kidney damage and drug metabolism related to cardiovascular diseases are not considered. Patent CN112618505A discloses a compound drug combining benazepril and pimoxandan to form a "white + brown" bilayer tablet. The conclusion of this compound drug is that its efficacy is non-inferior to that of the single-ingredient combination, indicating that this compound drug does not have a synergistic effect or reduce side effects.

[0006] Some research data indicate that most cardiovascular diseases in mammals are related to mitochondrial function in tissue cells. Nicotinamide adenine dinucleotide (NMN) has been extensively studied over the past century, proven to be an important coenzyme (coenzyme I) involved in many biochemical reactions within mammalian cells, serving as a "network dode" in many biological processes. Consequently, researchers worldwide have conducted extensive research on many precursors of NMN, particularly its direct precursor, nicotinamide mononucleotide (NMN), in areas such as anti-aging, diabetes, obesity, ischemia-reperfusion injury, heart failure (HF), cardiomyopathy, vascular dysfunction, cerebral hemorrhage, neuroprotection and cognitive function, Alzheimer's disease, retinal degeneration and corneal damage, acute kidney injury (AKI), and alcoholic liver disease. (Hong et al. reviewed Nicotinamide Mononucleotide: A Promising Molecule for Therapy of Diverse Diseases by Targeting NAD+Metabolism. Frontiers in Cell and Developmental Biology (2020) 8: Review Article 246). Among them, the most research on nicotinamide adenine dinucleotide supplement NMN is in the field of anti-aging health care, but its exact efficacy and anti-aging value at the tissue and organ level have not been convincingly explained in depth.

[0007] In the prior art, research on the protective effects of nicotinamide adenine dinucleotide (NMN) supplements in the field of heart disease includes the following: Takanobu Y et al. showed that NMN protects the heart from damage during ischemia or ischemia-reperfusion (Plos One, June 2014, Volume 9, Issue 6, e98972). Ke C et al. showed that NMN can inhibit cardiac dysfunction caused by Echs1 deficiency and may prevent the development of cardiomyopathy (JACC: Basic to Translational Science (2022) 7: 348-362). Studies have shown that high-dose (1000 mg / time * 2 times / day) supplementation with nicotinamide adenine dinucleotide supplement nicotinamide riboside (NR) can increase the level of nicotinamide adenine dinucleotide in the body, thereby improving mitochondrial function (JACC: Basic to Translational Science (2022) 7: 1183-1196). Studies have also shown that the deficiency of SIRT3, a nicotinamide adenine dinucleotide-dependent mitochondrial deacetylase, can lead to vasodilatory dysfunction, inflammation, and impaired mitochondrial respiration, promoting the development of atherosclerosis. Increased nicotinamide adenine dinucleotide levels can reduce plaque formation in blood vessels, improve vascular function, and reduce vascular inflammation. Joseph H et al. showed that heart failure with preserved ejection fraction (HFpEF) is associated with mitochondrial dysfunction and decreased nicotinamide adenine dinucleotide biosynthesis (Circulation Research (2021) 128: 1629-1641). The Buck Institute showed that NMN can reduce cardiac and systemic dysfunction caused by anthracycline chemotherapy drugs such as doxorubicin, and reduce their toxic side effects (Cells (2022) 12(1): 108). The Mayo Clinic showed that supplementing with NMN to increase nicotinamide adenine dinucleotide can prevent neurological damage and cognitive impairment caused by the chemotherapy drug cisplatin, and improve the quality of life of cancer survivors (Toxicology Letters (2021) 349: 115-123). Patent application CN113143946A discloses the protective effect of NMN against myocardial damage induced by anthracycline chemotherapy drugs. CN11375242A discloses the use of NMN in combination with a CD38 inhibitor (78C) in the preparation of a drug for the prevention or treatment of cardiotoxicity caused by azithromycin (one of the anthracycline chemotherapy drugs). CN117018009A discloses the pharmaceutical use of NMN in the preparation of a drug for the prevention or treatment of acute high-intensity exercise-induced myocardial injury. Earlier patent applications WO2016 / 188092A1 and CN106659729A also claimed the health care effects and pharmaceutical applications of NMN in arteriosclerosis and cardiovascular and cerebrovascular diseases caused by arteriosclerosis, but did not provide examples of equivalent or better drugs compared to existing drugs.

[0008] Observing the current research data on nicotinamide adenine dinucleotide (NMN) supplements in various disease areas, especially in cardiovascular diseases, it can be seen that NMN basically only exerts a certain protective (alleviating, reducing) effect on body tissues by increasing the level of nicotinamide adenine dinucleotide in the body. However, it is difficult to be certain that simply increasing nicotinamide adenine dinucleotide can cure or eradicate a certain heart-related disease. In other words, there are currently no reports of developing NMN or other nicotinamide adenine dinucleotide precursors, or even other mitochondrial function-enhancing substances, into drugs for the treatment of heart diseases.

[0009] There are also studies on the combined use of NMN with similar protective substances (melatonin, SS-31, exosomes, PQQ): Reza B et al. showed that the combined use of NMN and melatonin, which also has mitochondrial protective effects, enhanced the cardioprotective effect in aged rats (Molecular Biology Reports (2021)). Jeremy A et al. showed that NMN combined with a 50-fold dose of another mitochondrial protective targeting tetrapeptide, SS-31 (Elamipretide), improved cardiac function and reduced age-related cardiac hypertrophy (GeroScience (2022) 44: 1621-1639). Patent application CN116440162A discloses the combined use of NMN and exosomes extracted from the blood of young organisms in the field of anti-aging. However, there are no reports on the combined use of NMN with drugs for the treatment of heart disease.

[0010] Therefore, there is an urgent need to provide a pharmaceutical composition to solve the above problems. Summary of the Invention

[0011] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, the present invention proposes a pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases, which may have synergistic effects or reduce drug side effects in the prevention and / or treatment of mammalian cardiac diseases.

[0012] A pharmaceutical composition for the prevention and / or treatment of diseases of the mammalian cardiac system, comprising a mitochondrial function-optimizing substance and a therapeutic agent for heart-related diseases.

[0013] Furthermore, the mitochondrial function-enhancing substance includes at least one of oxidized nicotinamide adenine dinucleotide and oxidized nicotinamide adenine dinucleotide supplement.

[0014] Furthermore, the oxidized nicotinamide adenine dinucleotide supplement includes nicotinamide adenine dinucleotide precursors.

[0015] Furthermore, the nicotinamide adenine dinucleotide precursor includes at least one of DL-β-(3-indolyl)-α-aminopropionic acid, pyridine-3-carboxylic acid, 3-pyridinecarboxamide, nicotinamide ribose, and β-nicotinamide mononucleotide.

[0016] Furthermore, the mitochondrial function-optimizing substances also include 3,3′,4v,7-tetrahydroxyflavone, 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid, Omega-3 fish oil, N,(3-aminopropyl)-1,4-diaminobutane, 3-N-acetyl-5-methoxytryptamine, N-[2-(2-iodo-5-methoxyindol-3-yl)ethyl]acetamide, N-[2-(6-fluoro-5-methoxy-1H- At least one of the following: indole-3-yl)ethyl]acetamide, N-[2-(6-chloro-5-methoxy-1H-indole-3-yl)ethyl]acetamide, N-[2-(5-methoxy-2-phenyl-1H-indole-3-yl)ethyl]acetamide, (E)-5-[2-(4-hydroxyphenyl)-vinyl]-1,3-benzenediol, H2N-DArg-Dmt-Lys-Phe-NH2, ATP-promoting coenzymes, chlorophyll, B vitamins, curcumin, creatine, etc.

[0017] Furthermore, the therapeutic drug for heart-related diseases includes a pharmaceutical active ingredient selected from at least one of the following: positive inotropic drugs, angiotensin II receptor antagonists, platelet aggregation inhibitors, lipid-lowering drugs, antiarrhythmic drugs, angiotensin-converting enzyme inhibitors, diuretics, arterial vasodilators, and 5-HT antagonists.

[0018] Furthermore, the positive inotropic drug includes a phosphodiesterase inhibitor.

[0019] Furthermore, the positive inotropic drugs include at least one of pimoxendan, milrinone, amrinone, icariin, quinazoline, cilostazol, caffeine, aminophylline, 3-isobutyl-1-methylxanthine, paraxanthine, cyproterone acetate, theobromine, theophylline, vinpocetine, hydroxyindole, mesamidol, cyclolipan, isobutyllast, luteolin, drotaverine, piracetam, roflumilast, aprexalast, cetazidine, sildenafil, vardenafil, tadalafil, avanafil, udenafil, dipyridamole, papaverine, ammonia, cardiac glycosides, levosimendan, digitalis glycoside, digoxin, and strophanthidin K.

[0020] Furthermore, the angiotensin-converting enzyme inhibitors include at least one of captopril, cilazapril, quinalazapril, imidapril, benazepril, lisinopril, perindopril, quinapril, ramipril, enalapril, zolfenpril, and benazepril.

[0021] Furthermore, the angiotensin II receptor antagonist includes at least one of eprosartan, valsartan, irbesartan, flumasartan, azisartan, ozogartan, telmisartan, candesartan, losartan, and sacubitril / valsartan.

[0022] And / or,

[0023] The platelet aggregation inhibitors include at least one of cilostazol, aspirin, clopidogrel, trifluoromethanesulfonic acid, prasugrek, ticagrelor, troplotitin, tirofiban, aptifibatide, abciximab, vorapasal, tretroban, dipyridamole, ADP receptor antagonists, and GpIIb / IIIa receptor antagonists.

[0024] And / or,

[0025] The lipid-lowering drugs include at least one of simvastatin, atorvastatin, lovastatin, rosuvastatin, pravastatin, fluvastatin, pitavastatin, bezafibrate, probucol, acilimus, niacin, Src inhibitor Saracatinib, PCSK9 inhibitor Alirocumab, PCSK9 inhibitor Evolocumab, and myosin inhibitor Aficamten.

[0026] And / or,

[0027] The antiarrhythmic drugs include at least one of the following: digitalis, quinidine, procaine, disopyramide, phenytoin sodium, mexiletine, encaine, flecainide, propranolol, atenolol, diltiazem, amiodarone, metoprolol, propafenone, verapamil, lidocaine, ivabradine, bisoprolol, phentolamine, procainamide, ethoxymethazine, cefotaxime, acebutolol, and dicyclohexoperidine.

[0028] And / or,

[0029] The diuretic includes at least one of the following: amphotericin B, lithium citrate, tolvaptan, conivatan, dopamine, dazosulfan, acetazolamide, bumetanide, ethynyl acetylcholine, furosemide, torasemide, glucose, mannitol, eplerenone, spironolactone, amiloride, caffeine, theophylline, theobromine, benzylfluorothiazide, hydrochlorothiazide, cannic acid ester, and triamterene.

[0030] And / or,

[0031] The arterial vasodilators include at least one of adenosine, adrenaline, bradykinin, histamine, nicotinic acid, nitric oxide, diltiazem, verapamil, nifedipine, captopril, enalapril, hydralazine, pyrazinidine, acetylglycerol-ether-phosphocholine, prostacyclin, and isosorbide dinitrate.

[0032] And / or,

[0033] The 5-HT antagonist includes sargrelate, and at least one of fluoxetine, paroxetine, and sertraline.

[0034] Furthermore, the pharmaceutical composition comprises the nicotinamide adenine dinucleotide precursor, the positive inotropic drug, and / or the angiotensin-converting enzyme inhibitor and / or angiotensin receptor antagonist.

[0035] Furthermore, the pharmaceutical composition comprises at least one of oxidized nicotinamide adenine dinucleotide, 3,3′,4′,7-tetrahydroxyflavone, 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid, (E)-5-[2-(4-hydroxyphenyl)-vinyl]-1,3-benzenediol, H2N-DArg-Dmt-Lys-Phe-NH2, nicotinamide ribose, β-nicotinamide mononucleotide, DL-β-(3-indolyl)-α-aminopropionic acid, pyridine-3-carboxylic acid, 3-pyridinecarboxamide, pimoxendan, benazepril, telmisartan, benazepril hydrochloride, hydrochlorothiazide, sargrelate hydrochloride, sacubitril / valsartan, quinapril hydrochloride, enalapril, azilsartan, and furosemide.

[0036] The above-described embodiments of the present invention have at least one or more of the following beneficial effects:

[0037] The pharmaceutical composition for the prevention and / or treatment of cardiac diseases in mammals provided by this invention includes mitochondrial function-optimizing substances and drugs for treating cardiac diseases. Experimental verification has shown that the combined use of mitochondrial function-optimizing substances and drugs for treating cardiac diseases can have synergistic effects or reduce drug side effects in the prevention and / or treatment of cardiac diseases in mammals.

[0038] Furthermore, mitochondrial function-enhancing substances and drugs for treating heart-related diseases have a synergistic effect. Preparing mitochondrial function-enhancing substances with synergistic effects and drugs for treating heart-related diseases into a drug composition can reduce the frequency and amount of medication (e.g., number of pills), which significantly facilitates medication administration for pets and pet owners and reduces the difficulty of medication compliance.

[0039] Furthermore, substances that can be exogenously supplemented with cellular oxidized nicotinamide adenine dinucleotide (NDR) have the effects of protecting the heart from ischemia-reperfusion, promoting mitochondrial function to prevent heart failure, improving cardiac function, increasing stem cell survival rate, and reducing cellular oxidative stress. Their mechanism of action differs from existing heart disease treatments, and when combined with other drugs, they can exert a synergistic therapeutic effect. This coordinated preventative and therapeutic effect can reduce drug dosage, thereby lowering the probability of drug side effects, reducing tissue and cell damage, preventing or alleviating organ failure such as liver and kidney failure, and thus reducing the risk of treatment failure in animal heart diseases. During the occurrence and development of heart diseases, cardiomyocytes are easily damaged, and their function declines rapidly. Supplementing sufficient amounts of NDR with exogenously supplemented NDR can maintain bodily function. On the one hand, it can significantly reduce the risk or degree of cardiomyocyte damage, delaying the progression of the disease, thus reducing the amount of active ingredients in therapeutic drugs. On the other hand, it can alleviate the negative impact of the toxic side effects of active ingredients in therapeutic drugs on cells, thereby achieving a synergistic therapeutic effect. Conversely, without the presence of exogenous supplements of cellular oxidized nicotinamide adenine dinucleotide (Nd:A), the effectiveness of treatments for cardiovascular diseases will be reduced, and in severe cases, treatment may be lost altogether. Therefore, maintaining or increasing Nd:A levels in the body by utilizing exogenous supplements of Nd:A can help alleviate the toxic side effects of therapeutic drugs.

[0040] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0041] The disclosure of this invention will become more readily understood with reference to the accompanying drawings. It will be readily understood by those skilled in the art that these drawings are for illustrative purposes only and are not intended to limit the scope of protection of this invention.

[0042] Figure 1 This is a statistical chart of the in vitro test results of the synergistic effects of the pharmaceutical compositions in Examples 21-30 (F21-F30) of this application;

[0043] Figure 2 This is a statistical chart of the in vitro test results of the synergistic effects of the pharmaceutical compositions in Examples 31-40 (F31-F40) of this application;

[0044] Figure 3 This is a statistical chart of the in vitro test results of the pharmaceutical compositions of Examples 41-50 (F41-50) of this application synergistically reducing hepatotoxicity;

[0045] Figure 4This is a statistical chart showing the in vitro test results of the pharmaceutical compositions of Examples 51-60 (F51-60) of this application synergistically reducing hepatotoxicity;

[0046] Figure 5 This is a statistical chart of CK-MB detection data results from the in vivo synergistic study of the drug compositions in Examples 61-71 (F61-F71) of this application;

[0047] Figure 6 This is a statistical chart of LDH detection data results from the in vivo synergistic study of the pharmaceutical compositions in Examples 61-71 (F61-F71) of this application;

[0048] Figure 7 This is a statistical chart of SOD detection data results from the in vivo synergistic study of the pharmaceutical compositions in Examples 61-71 (F61-F71) of this application;

[0049] Figure 8 This is a statistical chart of the GSH-Px detection data results of the in vivo synergistic study of the drug compositions in Examples 61-71 (F61-F71) of this application. Detailed Implementation

[0050] Some embodiments of the present invention will now be described with reference to the accompanying drawings. Those skilled in the art should understand that these embodiments are merely illustrative of the technical principles of the present invention and are not intended to limit the scope of protection of the present invention.

[0051] To make the objectives and technical solutions of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings. Unless otherwise specified, the experimental methods described in the following embodiments are conventional methods; where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques and conditions described in the literature in the field or according to the product instructions; unless otherwise specified, the reagents and materials are commercially available. Any product identical or similar to this invention, derived by any person under the guidance of this invention or by combining features of this invention with other prior art, falls within the protection scope of this invention.

[0052] Of course, the embodiments used in this application are merely examples selected to illustrate the inventive concept and are not intended to limit the invention. Although the invention has been described in detail with reference to these embodiments, those skilled in the art can still modify the technical solutions described in the embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this invention should be included within the protection scope of this invention. The technical scope of this invention is not limited to the contents of the specification, but must be determined by the scope of the claims.

[0053] In this embodiment of the invention, the term "acceptable" means that the substance should possess characteristics that, taking into account the disease or condition to be treated and the respective route of administration, would prevent a rational and prudent medical practitioner from administering a substance to a patient that does not possess the "acceptable" characteristic. For example, for injectable substances, it is generally required that such substances be substantially sterile.

[0054] In embodiments of the present invention, the terms "therapeutic effective amount" and "effective amount" indicate that the substance and the amount of the substance are effective in preventing, alleviating or improving one or more symptoms of a disease or condition, and / or prolonging the survival of the subject receiving treatment.

[0055] The effective amount of each component to be applied, the biological half-life of the compound, the age, size, and weight of the subject, and any related medical conditions can be determined through standard procedures. The importance of these and other factors is well known to those skilled in the art.

[0056] It should be understood that combined use or co-administration includes use with other therapies, drugs, medical procedures, etc., wherein such other therapies or procedures may be administered at a time different from that of the compositions or components thereof of the present invention (e.g., within a short period of time (e.g., several hours, such as 1, 2, 3, 4-24 hours) or over a longer period of time (e.g., 1-2 days, 2-4 days, 4-7 days, 1-4 weeks) or at the same time as the compositions or components thereof of the present invention. Combined use also includes use with a therapy or medical procedure (e.g., surgery) that is administered once or infrequently, and accompanying administration of the compositions or components thereof of the present invention for a short or longer period of time before or after such other therapy or procedure. In some embodiments, the present invention is used to deliver the compositions or components thereof of the present invention and one or more other pharmaceutical therapeutic agents, delivered via the same or different routes of administration.

[0057] As the background art indicates, currently there is no single drug that can effectively and comprehensively treat cardiovascular diseases in companion animals and ornamental animals. Veterinarians often use multiple single drugs in combination, which involves not only the rational combination of drug types but also the proper control of dosage, as well as the pet owner's correct understanding and administration, and even the issue of animal compliance with complex and repeated medications. Furthermore, no existing drug combination can effectively address the treatment of cardiovascular diseases in companion animals and ornamental animals.

[0058] In this regard, the present invention creatively proposes a pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases, the pharmaceutical composition comprising mitochondrial function-optimizing substances and drugs for treating heart-related diseases, the pharmaceutical composition having synergistic or beneficial effects in the prevention and / or treatment of mammalian cardiac diseases.

[0059] Mitochondria are semi-autonomous organelles whose main functions include: synthesizing ATP, the organism's most basic energy source; assisting in ATP energy conversion; regulating calcium ion storage and balance, which are related to energy control; controlling apoptosis; and participating in cell signal transduction. Damage to or dysfunction of mitochondria can lead to a range of diseases, and normal mitochondrial function is crucial for human health.

[0060] Mitochondrial function-enhancing substances are substances that can increase the number or content of mitochondria. Although mitochondrial function-enhancing substances cannot directly treat heart-related diseases themselves, or completely eliminate or alleviate the symptoms caused by heart-related diseases, this application has found through research that mitochondrial function-enhancing substances can be formulated into pharmaceutical compositions with drugs for treating heart-related diseases for the prevention and / or treatment of mammalian heart diseases. Mitochondrial function-enhancing substances have a synergistic effect with drugs for treating heart-related diseases, and their efficacy is superior to that of existing combination formulations or single-agent formulations of the same drug at the same dosage. Furthermore, they can reduce the severity or probability of side effects to a level lower than that of existing combination formulations or single-agent formulations of the same drug at the same dosage.

[0061] Furthermore, the mitochondrial function-optimizing substance includes oxidized nicotinamide adenine dinucleotide (NAD). + At least one of the following: oxidized nicotinamide adenine dinucleotide supplement.

[0062] Oxidized nicotinamide adenine dinucleotide (NAD) + It is closely related to the energy production process of mitochondria, and oxidized nicotinamide adenine dinucleotide (NAD) + A decrease in mitochondrial levels can adversely affect mitochondrial function, leading to metabolic dysfunction.

[0063] Oxidized nicotinamide adenine dinucleotide (NAD) supplements are classified into direct and indirect supplements based on their mechanism of action. Among them, DL-β-(3-indolyl)-α-aminopropionic acid (Trp), pyridine-3-carboxylic acid (NA), 3-pyridinecarboxamide (NAM), nicotinamide ribose (NR), and β-nicotinamide mononucleotide (NMN) are examples of direct supplements of oxidized nicotinamide adenine dinucleotide. Studies have shown that the PARP inhibitor AZD2281 (olaparib) can promote the production of nicotinamide adenine dinucleotide (NAD) in nematodes. + ) levels are increased, similar to AZD2281, which can promote the increase of nicotinamide adenine dinucleotide (NAD) levels in the body. +The substance at the ) level is nicotinamide adenine dinucleotide (NAD). + Indirect supplement.

[0064] The oxidized nicotinamide adenine dinucleotide supplement includes a nicotinamide adenine dinucleotide precursor. The nicotinamide adenine dinucleotide precursor is a substance that can be directly or indirectly converted into oxidized nicotinamide adenine dinucleotide in mammals, or a substance that can directly or indirectly increase the level of nicotinamide adenine dinucleotide in mammals.

[0065] Nicotinamide adenine dinucleotide (NADP) precursors have many advantages, such as virtually unlimited tolerable dose limits (substances like 3-pyridinecarboxamide, nicotinamide riboside, and β-nicotinamide mononucleotide are widely present in and outside organisms, posing no biocompatibility issues), and virtually no adverse effects on absorption, distribution, or metabolism (the related substances involved in their metabolism are also commonly found in the body). Besides synergistically enhancing the effects of medications for cardiovascular diseases, supplementing with NADP can also provide many other beneficial protective and health-promoting effects. The aforementioned NADP and its multiple precursors are all widely present in nature (including in various organisms). Among them, β-nicotinamide mononucleotide, the most direct precursor to NADP, is the preferred exogenous form of cellular oxidized NADP, essentially a nucleotide with a β-N-glycosidic bond at the C-1 position of the nitrogenous base nicotinamide and D-ribose. Oral administration is currently the most common and safe method. The WHO recommends an upper limit of 900 mg per day for β-nicotinamide mononucleotide. Further animal studies in rats suggest that the upper limit of β-nicotinamide mononucleotide can be increased to 1500 mg / kg / day, which also demonstrates that nicotinamide adenine dinucleotide and its precursors have adequate safety.

[0066] Specifically, the nicotinamide adenine dinucleotide precursor includes at least one of DL-β-(3-indolyl)-α-aminopropionic acid (Trp), pyridine-3-carboxylic acid (NA), 3-pyridinecarboxamide (NAM), nicotinamide ribose (NR), and β-nicotinamide mononucleotide (NMN). Among them, 3-pyridinecarboxamide (NAM) has the chemical formula C6H6N2O, a molecular weight of 122.125, and CAS: 98-92-0; nicotinamide ribose (NR) has the chemical formula C... 11 H 15 N2O5, molecular weight 255.247, CAS: 1341-23-7; β-nicotinamide mononucleotide (NMN) chemical formula is C 11 H 15 N 2O8P, with a molecular weight of 334.219, CAS: 1094-61-7.

[0067] In addition to the oxidized nicotinamide adenine dinucleotide (NAD) mentioned above + In addition to oxidized nicotinamide adenine dinucleotide supplements, mitochondrial function-optimizing substances include 3,3′,4′,7-tetrahydroxyflavone, 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid, Omega-3 fish oil, N,(3-aminopropyl)-1,4-diaminobutane, 3-N-acetyl-5-methoxytryptamine, N-[2-(2-iodo-5-methoxyindol-3-yl)ethyl]acetamide, N-[2-(6-fluoro-5-] [Methoxy-1H-indol-3-yl)ethyl]acetamide, N-[2-(6-chloro-5-methoxy-1H-indol-3-yl)ethyl]acetamide, N-[2-(5-methoxy-2-phenyl-1H-indol-3-yl)ethyl]acetamide, (E)-5-[2-(4-hydroxyphenyl)-vinyl]-1,3-benzenediol, H2N-DArg-Dmt-Lys-Phe-NH2, ATP-promoting coenzymes, chlorophyll, B vitamins, curcumin, creatine, etc.

[0068] Preferably, the mitochondrial function-enhancing substance includes H2N-DArg-Dmt-Lys-Phe-NH2 and 3-N-acetyl-5-methoxytryptamine, wherein the chemical formula of H2N-DArg-Dmt-Lys-Phe-NH2 is C 32 H 49 N 9O5 With a molecular weight of 639.39 and CAS number 736992-21-5, the chemical formula of 3-N-acetyl-5-methoxytryptamine is C2. 13 H 16 N 2O2 The molecular weight is 232.278, CAS: 73-31-4. The chemical formula of 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid is C. 14 H6N 2O8 The molecular weight is 330.206, and the CAS number is 72909-34-3.

[0069] The medication for treating heart-related diseases includes a pharmaceutically active ingredient selected from at least one of the following: positive inotropic drugs, angiotensin II receptor antagonists, platelet aggregation inhibitors, lipid-lowering drugs, antiarrhythmic drugs, angiotensin-converting enzyme inhibitors, diuretics, arterial vasodilators, and 5-HT antagonists.

[0070] The positive inotropic drugs include phosphodiesterase inhibitors. Further, positive inotropic drugs such as phosphodiesterase inhibitors include at least one of pimoxendan, milrinone, amrinone, icariin, quinazoline, cilostazol, caffeine, aminophylline, 3-isobutyl-1-methylxanthine, paraxanthine, pentoxyfilline, theobromine, theophylline, vinpocetine, hydroxyindole, mesamidol, cyclolipan, isobutyllast, luteolin, drotavirin, piramylast, roflumilast, apralast, cisaborole, sildenafil, vardenafil, tadalafil, avanafil, udenafil, dipyridamole, papaverine, ammonia, cardiac glycosides, and levosimendan.

[0071] Preferably, the positive inotropic drug is selected from at least one of pimoxendan, cilostazol, roflumilast, and levosimendan. Roflumilast has the chemical formula C0. 17 H 14 C 12 F2N 2O3 The molecular weight is 403.21, and the CAS number is 162401-32-3.

[0072] Furthermore, the angiotensin II receptor antagonist includes at least one of eprosartan, valsartan, irbesartan, flumasartan, azisartan, olmesartan, telmisartan, candesartan, losartan, and sacubitril / valsartan (LCZ696).

[0073] Preferably, the angiotensin II receptor antagonist is telmisartan. The chemical formula of telmisartan is C2. 33 H 30 N 4O2 The molecular weight is 514.237, and the CAS number is 144701-48-4.

[0074] Furthermore, the platelet aggregation inhibitors include at least one of cilostazol, aspirin, clopidogrel, trifluoromethanesulfonic acid, prasugrek, ticagrelor, toclopidine, tirofiban, aptifibatide, abciximab, vorapasal, trutroban, dipyridamole, ADP receptor antagonists, and GpIIb / IIIa receptor antagonists.

[0075] Preferably, the platelet aggregation inhibitor is cilostazol or tirofiban.

[0076] Furthermore, the lipid-lowering drugs include at least one of simvastatin, atorvastatin, lovastatin, rosuvastatin, pravastatin, fluvastatin, pitavastatin, bezafibrate, probucol, acilimus, niacin, Src inhibitor Saracatinib (AZD0530), PCSK9 inhibitor Alirocumab, PCSK9 inhibitor Evolocumab, and myosin inhibitor Aficamten (CK-274).

[0077] Preferably, the lipid-lowering drug includes niacin and probucol. Probucol has the chemical formula C0. 31 H 48O2 S2, molecular weight 516.84, CAS: 23288-49-5.

[0078] Furthermore, the antiarrhythmic drugs include at least one of digitalis preparations, quinidine, procaine, disopyramide, phenytoin sodium, mexiletine, encaine, flecainide, propranolol, atenolol, diltiazem, amiodarone, metoprolol, propafenone, verapamil, lidocaine, ivabradine, bisoprolol, phentolamine, procainamide, ethoxymethazine, cefotaxime, acebutolol, and dicyclohexoperidine.

[0079] Preferably, the antiarrhythmic drug includes propranolol, lidocaine, phenytoin sodium, and acebutolol. Among them, the chemical formula of acebutolol is C1. 18 H 28 N 2O4 The molecular weight is 336.43, and the CAS number is 37517-30-9.

[0080] Furthermore, the angiotensin-converting enzyme inhibitors include at least one of captopril, cilazapril, quinalazapril, imidapril, benazepril, lisinopril, perindopril, quinapril, ramipril, enalapril, zolfenpril, and benazepril.

[0081] Preferably, the angiotensin-converting enzyme inhibitor includes benazepril, enalapril, and quinapril. Among them, the chemical formula of quinapril is C2. 25 H 30 N 2O5 The molecular weight is 438.516, and the CAS number is 85441-61-8.

[0082] Furthermore, the diuretic includes at least one of amphotericin B, lithium citrate, tolvaptan, conivovatan, dopamine, dazosulfan, acetazolamide, bumetanide, ethnocyanidin, furosemide, torasemide, glucose, mannitol, eplerenone, spironolactone, amiloride, caffeine, theophylline, theobromine, benzylfluorothiazide, hydrochlorothiazide, and canonical esters. The chemical formula of hydrochlorothiazide is C7H8C1N. 3O4 S2, with a molecular weight of 297.739, CAS 58-93-5.

[0083] Furthermore, the arterial vasodilator includes at least one of adenosine, adrenaline, bradykinin, histamine, nicotinic acid, nitric oxide, diltiazem, verapamil, nifedipine, captopril, enalapril, hydralazine, pyrazinidine, acetylglycerol-ether-phosphocholine (platelet-activating factor), prostacyclin, and isosorbide dinitrate.

[0084] Preferably, the arterial vasodilator includes nicotinic acid, enalapril, and captopril. Captopril has the molecular formula C9H10. 15 NO3S, molecular weight 217.285, CAS: 62571-86-2.

[0085] Furthermore, the 5-HT antagonist includes sargrelate, and at least one of fluoxetine, paroxetine, and sertraline. Sargrelate has the molecular formula C5. 24 H 31 NO6, molecular weight 429.506, CAS 125926-17-2.

[0086] Optionally, the medication used to treat veterinary heart diseases can be at least one cardiotonic drug with positive inotropic effects, such as pimoxendan, digitoxin, digoxin, strophanthidin K, etc.; or at least one angiotensin inhibitor, such as benazepril, enalapril, captopril, etc.; or one or more of cardiotonic drugs and angiotensin inhibitors.

[0087] Alternatively, the medication used to treat veterinary heart disease may be at least one of the above-mentioned medications combined with a diuretic substance, such as furosemide, hydrochlorothiazide, or triamterene.

[0088] Optionally, the pharmaceutical composition includes the nicotinamide adenine dinucleotide precursor, the positive inotropic drug, and / or the angiotensin-converting enzyme inhibitor and / or angiotensin receptor antagonist.

[0089] Optionally, the pharmaceutical composition comprises at least one of oxidized nicotinamide adenine dinucleotide, 3,3′,4′,7-tetrahydroxyflavone, 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid, (E)-5-[2-(4-hydroxyphenyl)-vinyl]-1,3-benzenediol, H2N-DArg-Dmt-Lys-Phe-NH2, nicotinamide ribose, β-nicotinamide mononucleotide, DL-β-(3-indolyl)-α-aminopropionic acid, pyridine-3-carboxylic acid, 3-pyridinecarboxamide, pimoxendan, benazepril, telmisartan, benazepril hydrochloride, hydrochlorothiazide, sargrelate hydrochloride, sacubitril / valsartan, quinapril hydrochloride, enalapril, azilsartan, and furosemide.

[0090] Scientifically selecting drugs with different metabolic and elimination pathways to form drug combinations helps prevent the accumulation of side effects and reduces the risk of adverse reactions to organs such as the liver and kidneys. For example, pimoxemandil, with its positive inotropic and calcium ion sensitizing mechanism of action, is mainly excreted through feces, while benazepril, with its mechanism of action of inhibiting angiotensin-converting enzyme and kininase, is excreted through the liver, gallbladder, and kidneys. Furthermore, the combined use of pimoxemandil and benazepril provides a broader spectrum of treatment for various types of veterinary heart diseases, making medication administration more convenient for veterinarians and pet owners.

[0091] It should be noted that the therapeutic drugs for heart-related diseases in the embodiments of the present invention include active pharmaceutical ingredients. The active pharmaceutical ingredients are selected from various possible derivatives of the following without altering their active properties, such as phosphodiesterase inhibitors and other positive inotropic drugs, and / or angiotensin II receptor antagonists, and / or platelet aggregation inhibitors, and / or lipid-lowering drugs, and / or angiotensin-converting enzyme inhibitors, and / or diuretics, and / or arterial vasodilators, and / or 5-HT antagonists, or their various salt-forming forms. The active pharmaceutical ingredients are also included in various possible crystalline forms and existing forms.

[0092] All of the above-mentioned optional technical solutions can be combined in any way to form the optional embodiments of this application, and will not be described in detail here.

[0093] The beneficial effects of this application will be further illustrated below with reference to the embodiments.

[0094] The raw materials used in the embodiments and comparative examples are described below:

[0095] In Tables 1, 2, 3, and 4, A refers to mitochondrial function-optimizing substances, and B refers to drugs for treating heart-related diseases. In Table 1, F01-F20 refer to Examples 1 to 20; in Table 2, F21-F40 refer to Examples 21 to 40; in Table 3, F41-F60 refer to Examples 41 to 60; and in Table 4, F61-F71 refer to Examples 61 to 71. F01 refers to a pharmaceutical composition composed of substances A07 and B01, and F11 refers to a pharmaceutical composition composed of substances A05, A07, B01, B07, and B08.

[0096] In Table 1, A01: oxidized nicotinamide adenine dinucleotide, A02: 3,3′,4′,7-tetrahydroxyflavone, A03: 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid, A04: (E)-5-[2-(4-hydroxyphenyl)-vinyl]-1,3-benzenediol, A05: H2N-DArg-Dmt-Lys-Phe-NH2, A06: nicotinamide ribose, A07: β-nicotinamide mononucleotide, A08: D L-β-(3-indolyl)-α-aminopropionic acid, A09: pyridine-3-carboxylic acid, A10: 3-pyridinecarboxamide, B01: pimoxendan, B02: levosimendan, B03: telmisartan, B04: clopidogrel, B05: probucol, B06: acebutolol hydrochloride, B07: benazepril hydrochloride, B08: hydrochlorothiazide, B09: captopril, B10: sarpogrelate hydrochloride, B11: cilostazol, B12: tirofiban hydrochloride, B13: lidocaine, B14: diltiazem hydrochloride. √ indicates that the corresponding material participated in the experimental implementation of this group of drug compositions.

[0097] In Table 2, the substances corresponding to A01-A10 are the same as those corresponding to A01-A10 in Table 1. B01: Pimobendan, 1.25 mg / L; B11: Cilostazol, 0.5 μmol / L; B02: Levosimendan, 0.1 μmol / L; B15: Verapamil, 1 mg / L; B16: Eprosartan Mesylate; B03: Telmisartan, 1 mg / L; B17: Valsartan, 50 mM; B18: Amlodipine Besylate; B19: Sacubitril; B20: Atorvastatin Calcium; B21: Cicatinib AZD0530; B22: CK-274; B23: Propranolol; B24: Ivabradine; B04: Clopidogrel Bisulfate; B10: Sarpogrelate; B07: Benazepril; B25: Enalapril Maleate; B08: Hydrochlorothiazide. √ indicates that the corresponding material participated in the experimental implementation of this group of drug compositions.

[0098] In Table 3, the substances corresponding to A01-A10 are the same as those corresponding to A01-A10 in Table 1. B11: Cilostazol 0.5 μmol / L, B01: Pimobendan 25 mg / L, B02: Levosimendan >0.3 μmol / L, B26: Cardiac glycoside 125 mg / L, B27: Irbesartan 20 μmol / L, B03: Telmisartan 10 mg / L, B28: Sacubitril / Valsartan, LCZ696, 500 mM / L, B29: Quinidine, B30: Atenolol, B31: Bumetanide, B32: Hydralazine, B33: Diltiazem Hydrochloride, B34: Paroxetine Hydrochloride, B35: Alirocumab, B10: Sarpogrelate, B36: Saracatinib, AZD0530, B37: Clopidogrel Sulfate, B38: Quinapril Hydrochloride. √ indicates that the corresponding material participated in the experimental implementation of this group of drug compositions.

[0099] In Table 4, the substances corresponding to A01-A10 are the same as those corresponding to A01-A10 in Table 1. B01: Pimobendan, B03: Telmisartan, B07: Benazepril Hydrochloride, B08: Hydrochlorothiazide, B10: Sarpogrelate Hydrochloride, B28: Sacubitril / Valsartan, LCZ 696, 500 mM / L, B38: Quinapril Hydrochloride, B39: Enalapril, B40: Azilsartan, B41: Furosemide. In Examples 61-71, the dosages of each Class A and Class B substance in each pharmaceutical composition were half (1 / 2) of the dosages of each Class A and Class B substance in the corresponding Class A and Class B substance treatment groups, respectively. √ indicates that the corresponding material participated in the experimental implementation of that group of pharmaceutical compositions.

[0100] Performance testing methods:

[0101] 1. Feasibility and stability testing of drug compositions

[0102] Feasibility studies and assessments of relevant drug compositions were conducted using conventional pharmaceutical methods. Based on the physicochemical properties of various mitochondrial function-enhancing substances and cardiovascular disease treatment drugs involved in the experiments, such as solubility, melting point, and susceptibility to oxidation, appropriate and acceptable pharmaceutical excipients and formulations were adopted.

[0103] Accelerated stability studies were conducted on the pharmaceutical compositions prepared in Examples 1-20 according to the methods in ICH Guidelines Q1: Stability (Q1A(R): Stability Testing of New Drugs and Products). The test conditions were: temperature 30±2℃, relative humidity 60%±5%, and sampling and testing at 0, 1, 2, 3, and 6 months.

[0104] Among them, Table 1 ■This embodiment of the experiment includes consideration of the expected route of administration, release mode / rate, and targeting trend of the drug composition, the use of acceptable excipients and formulations, and consideration of necessary incompatibilities. It includes the use of pharmaceutical processing methods such as micronization, solubilization, and emulsification when necessary; the use of pharmaceutical technologies such as encapsulation, nanocapsules, sustained release, water-in-oil / oil-in-water / oil-in-oil; and the use of acceptable excipients appropriate to each relevant dosage form. S refers to solid forms such as powders, tablets, and ointments; L refers to liquid or semi-liquid forms such as solutions, suspensions, and emulsions; and I refers to the form used for injection. ● indicates the form of the drug composition implemented in the corresponding group, and signifies that the form of the drug composition is operable and can achieve acceptable drug administration; ★ indicates that the accelerated stability study results of the drug composition implemented in the corresponding group meet the stability standards and principles described in the ICH guidelines.

[0105] 2. In vitro synergistic effect test of drug composition

[0106] Establishment of a modeling substance for a cardiac disease state: Raw264.7 macrophage culture medium was stimulated with LPS to obtain Raw264.7 macrophage supernatant containing pro-inflammatory factors such as NO, TNF-α, IL-1β, and IL-6. This modeling substance, referred to as the modeling substance, was prepared to simulate the "cardiac disease" state environment in which the cells were located. The effectiveness of the modeling substance was assessed by culturing H9c2 cells (with and without the modeling substance) and detecting inflammatory factors in the cell cultures.

[0107] Establishing an in vitro cell model of cardiac disease under cardiac system disease conditions: using H9c2 cells at 1-5*10 4 Cells / well were cultured in the same adherent culture medium and starved for 12 hours at the same time point. An in vitro cell model of cardiac disease was established by adding the above-mentioned modeling material, and the success of the model was confirmed by detecting cell viability.

[0108] Using the CTG method (Promega) The Luminescent Cell Viability Assay (CTG) detects cell viability using an ATP-based method to determine the number of viable cells in a culture. It employs a highly efficient "sample loading-mixing-measurement" system. The detection reagent has a long signal duration, making this method widely used for cell viability detection and internationally recognized for its high accuracy. CellTiter-Glo was added to a 96-well plate. TMThe reagent solution was left to stand at room temperature for 10 minutes before the fluorescence value was detected using an ELISA reader. A standard curve was first constructed using the same conditions as the healthy control group: culture medium with a cell concentration 10 times that of the healthy control group was serially diluted to single-digit cell concentration levels, and cell counts were measured. A standard curve was constructed based on the correlation between cell concentration data and measured fluorescence values, and the recovery rate was calculated to validate the detection curve.

[0109] Synergy testing of the drug composition: Batch experiments were conducted using 96-well plates, with wells divided into three groups (F21-F40): "healthy control group without modeling material", "positive disease control group with modeling material", and "treatment test group with modeling material and drug". Each treatment test group was a synergy testing experiment involving a drug composition, including the drug composition "treatment", the mitochondrial optimization substance "treatment" in the group, and the cardiac system treatment drug "treatment" in the group. Healthy H9c2 cells in each group were treated according to the above-described in vitro cell modeling method until step "starvation for 12 h". Then, the effective modeling material for cardiac disease states obtained by the above method was added in equal amounts or proportions (the healthy control group received an equal amount of PBS buffer instead of the modeling material, and the positive disease control group received PBS buffer instead of the "drug" for sham treatment). Simultaneously, according to the experimental design, the corresponding "conventional treatment dose" of treatment material (drug composition, drug composition A-type substance, drug composition B-type substance) was added to the corresponding wells. The cells were cultured in the same environment for another 24 h, and cell viability was detected and compared among the groups.

[0110] Data processing and synergy analysis: One-way ANOVA was used to compare means among multiple groups, and two-tailed t-tests were used to compare means between two groups. cvAB represents the cell viability value after "treatment" of each composition, cvA represents the cell viability value after "treatment" of substance A in each drug composition, and cvB represents the cell viability value after "treatment" of substance B in each drug composition. P < 0.05 indicates statistical difference, and P < 0.01 indicates significant statistical difference. Experimental results were categorized into three types: ★ (synergistic effect, i.e., cvAB > cvA, cvAB > cvB, and P < 0.05 or 0.01), ☆ ("not inferior", i.e., cvAB > cvA, cvAB > cvB, and P ≥ 0.05), and ↓ (ineffective or "inferior", i.e., cvAB > cvA, cvAB > cvB, and P ≥ 0.05). <cvA or cvB)。

[0111] 3. Test on the synergistic reduction of drug toxicity by drug compositions

[0112] A modeling substance for cardiac disease states was established (using the same method as "2. In vitro test of synergistic effect of drug composition" for "establishing a modeling substance for cardiac disease states"). The effectiveness of the modeling substance was tested by culturing human renal cells HEK293 (culture group with modeling substance added and blank control group without modeling substance added) and detecting inflammatory factors in the cell culture.

[0113] Establishing an in vitro kidney cell model under cardiac disease conditions: using HEK293 cells at 1-5*102 6 Cells / well were cultured in the same adherent culture medium and starved for 12 hours at the same time point. An in vitro cell model of cardiac disease was established by adding the above-mentioned modeling material, and the success of the model was confirmed by detecting cell viability.

[0114] Cell viability in each experimental group was detected using the CTG method (the same method used in "2. In vitro test of synergistic effect of drug composition" for "detecting cell viability using the CTG method").

[0115] Synergistic Reduction of Drug Toxicity Test by Composition: Batch experiments were conducted using 96-well plates, with wells divided into three groups (F41-F60): "Health Control Group without Modeling Material", "Positive Disease Control Group with Modeling Material", and "Treatment Experiment Group with Modeling Material and Drug". The treatment experiment group (e.g., F41) was further divided into "treatment" comparison experiments for various compositions as designed. Each composition "treatment" comparison experiment specifically included "composition treatment" and "treatment with therapeutic drug". Healthy H9c2 cells of equal quantity or proportion in each group were treated according to the above-described in vitro cell modeling method until step "starvation for 12 hours". Then, the effective modeling material for cardiac disease states obtained by the above method was added in equal quantity or proportion (the healthy control group received an equal volume of PBS buffer instead of the modeling material, and the positive disease control group received PBS buffer instead of the "drug" for sham treatment). Simultaneously, according to the experimental design, the corresponding "toxic dose" of treatment material (single component or composition of the composition) was added to the corresponding wells. The cells were cultured in the same environment for another 24 hours, and cell viability was detected and compared among the groups.

[0116] Data processing and toxicity reduction analysis: Two-tailed t-tests were used to compare the means between the two groups. P < 0.05 indicated a statistically significant difference, and P < 0.01 indicated a statistically significant difference. sAB represents the cell viability (Survival) value after "treatment" with each drug composition, sA represents the cell survival rate after "treatment" with substance A in each composition, and sB represents the cell viability value after "treatment" with substance B in each composition. Experimental results were categorized into three types: (It has a synergistic effect in reducing toxic side effects, i.e., sAB>sB, and P<0.05 or 0.01). ("No increase in toxicity" effect, i.e., sAB>sB, and P≥0.05), ↓ (ineffective or "increase in toxicity" effect, i.e., sAB≤sB).

[0117] 4. In vivo synergistic testing of drug compositions

[0118] Experimental grouping: Male mice aged 7-8 weeks and weighing 20±5g were divided into experimental groups, including a negative control group (i.e., normal group N), a positive control group (i.e., model group M), and treatment comparison experimental groups F61-F71 for each example. The treatment comparison experimental groups for each example further included a drug composition treatment group (A+B), a mitochondrial function optimization substance treatment group (A), and a heart-related disease treatment drug treatment group (B).

[0119] A mouse model of cardiac disease was established: Mice in the positive control group (M) and the treatment experimental group were intraperitoneally injected with isoproterenol hydrochloride (150 mg / kg) on ​​the first day, followed by a second injection (150 mg / kg) 24 hours later on the second day. Experiments began on the third day after another 24 hours. Mice in the negative control group (N) were intraperitoneally injected twice with an equal volume of physiological saline.

[0120] Experimental treatments for each group: Mice in groups N and M were simultaneously administered an equal volume of physiological saline as a drug replacement by gavage daily for 7 consecutive days (days 3 to 9). On day 10, mice were fasted for 12 hours before sampling, but were allowed free access to food and water during the remaining days. In each example, the treatment experimental groups were administered the corresponding therapeutic drug composition (A+B), mitochondrial function optimizing substance (A), and heart-related disease treatment drug (B) by gavage for 7 consecutive days. On day 10, mice were fasted for 12 hours before sampling, but were allowed free access to food and water during the remaining days. For example, in the F71 treatment comparison experimental group, the drug composition treatment group was administered the composition (A03+A05+A07+B01+B40+B41) by gavage, the heart-related disease treatment drug treatment group was administered the corresponding substance A (A03+A05+A07) by gavage, and the mitochondrial function optimizing substance treatment group was administered the corresponding substance B (B01+B40+B41) by gavage.

[0121] Treatment efficacy detection and statistical analysis: In clinical practice, plasma creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) are important diagnostic indicators of cardiac function (myocardial ischemia), while plasma or serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) are important evaluation indicators of the body's antioxidant defense system. Blood samples were collected from mice in each group, and these biochemical indicators were detected and compared to evaluate the differences in efficacy between the treatment control groups. GraphPadPrism 8 (GraphPad Software, USA) was used for statistical analysis of the data. All data are presented as mean ± standard mean error (mean ± SD), and at least three independent experiments were ensured for each group. One-way ANOVA was used for comparison of means among multiple groups, and two-tailed t-tests were used for pairwise comparison of means between groups. p < 0.05 indicated statistical difference (*), and p < 0.01 or 0.005 indicated significant statistical difference (** / ***).

[0122] Synergy Analysis and Judgment: In this set of examples, synergistic effect refers to the combined effect of mitochondrial function-optimizing substances and drugs for treating heart-related diseases being greater than the sum of the effects of using these two substances individually. The Model-Free Test (MFT) method was used to evaluate the synergistic effect. The drug interaction index γ = a / A + b / B, where A and B are the doses that produce the same effect when the two substances are used individually and in combination, and a and b are the individual doses of the two substances when used in combination. γ > 1 indicates that the two substances are antagonistic, γ = 1 indicates that the interaction between the two substances is additive, and γ < 1 indicates that the two substances are synergistic.

[0123] The test results of the pharmaceutical compositions prepared in Examples 1-20 are shown in Table 1; the test results of the pharmaceutical compositions prepared in Examples 21-40 are shown in Table 2; the test results of the pharmaceutical compositions prepared in Examples 41-60 are shown in Table 3; and the test results of the pharmaceutical compositions prepared in Examples 61-71 are shown in Table 4.

[0124] Table 1 Results of feasibility and stability tests of the drug composition

[0125]

[0126]

[0127] Table 2. Results of in vitro tests for synergistic effects of drug compositions.

[0128]

[0129]

[0130] Table 3 Results of the test on the synergistic reduction of drug toxicity by drug compositions

[0131]

[0132] Table 4 Results of in vivo synergistic testing of drug compositions

[0133]

[0134]

[0135] Test results:

[0136] 1. As can be seen from the test results in Table 1, the pharmaceutical compositions prepared using mitochondrial function-optimizing substances and drugs for treating heart-related diseases provided in this application can generally be processed by conventional pharmaceutical methods to form pharmaceutically acceptable compound drugs or compositions, and can generally meet the acceptable stability standards for pharmaceutical applications.

[0137] 2. As shown in Table 2, the combined use of Class A mitochondrial function-enhancing substances and Class B cardiovascular disease treatment drugs synergistically increases cell viability in damaged cell models (shown as ★ in most examples), indicating that the combination of mitochondrial function-enhancing substances and Class B cardiovascular disease treatment drugs to form a drug composition has superiority.

[0138] 3. As shown in Table 3, the combined application of mitochondrial function-optimizing substances and drugs for treating heart-related diseases synergistically improves the cell survival rate of the HEK293 damaged human kidney cell model (all showed synergistic effects). This indicates that the combined use of Class A mitochondrial function-enhancing substances and Class B therapeutic drugs has a synergistic effect in reducing hepatotoxicity and nephrotoxicity.

[0139] 4. As shown in Table 4, the combined use of the mitochondrial function-optimizing substance and the drug for treating heart-related diseases has a synergistic effect (mostly significant differences P<0.01 or 0.005, a few examples with statistically significant differences P<0.05) in reducing CK-MB and LDH levels and increasing SOD and GSH-Px levels in vivo. It also has a synergistic (all Y<1) effect in protecting the heart and kidneys and treating heart-related diseases. Therefore, from an in vivo perspective, this invention's pharmaceutical composition demonstrates that it is non-inferior to or has a synergistic effect compared to using the mitochondrial function-optimizing substance and the drug for treating heart-related diseases separately.

[0140] In this specification, "pharmaceutical composition," "compound drug," or other similar terms or names refer to compositions comprising medicaments for the treatment and / or diagnosis and / or cure and / or prevention of disease. Furthermore, a drug means any substance or combination of substances (composition) that exhibits properties of treating or preventing disease in humans or animals; or any substance or combination of substances that may be used or administered to humans for the purpose of restoring, correcting, or altering physiological function or for medical diagnosis by exerting pharmacological, immunological, or metabolic effects. According to the FDA glossary, in the context of this invention, "compound drug," "pharmaceutical composition," "drug mixture," or "composition" also means "pharmaceutical product," which is a final dosage form that typically contains a pharmaceutical substance but not necessarily combined with other active or inactive ingredients. A drug is defined as a substance recognized by an official pharmacopoeia or prescription, a substance intended for the diagnosis, cure, relief, treatment, or prevention of disease, a substance intended to affect the structure or any function of the body (excluding food), or a substance intended to be used as a component of a drug rather than a device or a component, part, or accessory of a device (biological products are included in this definition and are generally covered by the same laws and regulations, but differ regarding their manufacturing process—chemical processes versus biological processes). According to the present invention, the term "veterinary drug" has the same definition as "medicine," but applies to animals (meaning non-humans): "animal" means any living member of the animal kingdom other than humans. More precisely, "veterinary drug" (or medicine or composition) means any substance or mixture of substances used, manufactured, sold, or manifested as suitable for the diagnosis, treatment, control, elimination, relief, or prevention of disease or abnormal physiological or mental state or symptoms thereof in animals; or for the restoration, correction, control, or alteration of any physiological, mental, or organic function in animals.

[0141] In embodiments of the present invention, the pharmaceutical composition comprises preparing the pharmaceutical composition referred to in the present invention into any acceptable and suitable dosage form, depending in part on the use or route of administration, such as oral, transdermal, transmucosal, inhalation, or injection (parenteral). The composition or components thereof can be administered via various routes, including intravenous, intraperitoneal, subcutaneous, intramuscular, oral, transmucosal, rectal, transdermal, or inhalation. In some embodiments, oral administration and associated dosage forms are preferred; for example, the compound can be formulated into conventional oral dosage forms, such as capsules, tablets, and liquid formulations, such as syrups, elixirs, and concentrated drops.

[0142] Pharmaceutical formulations for oral use can be obtained, for example, by combining a pharmaceutical composition or its components with a solid excipient, optionally by grinding a mixture, and then processing the mixture into granules (if necessary) after adding suitable excipients, thereby obtaining tablets or sugar-coated pills. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose formulations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose (CMC), and / or polyvinylpyrrolidone (PVP: povidone)). If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginate or their salts, such as sodium alginate, can be added.

[0143] Alternatively, injection (parenteral administration) can be used, such as intramuscular, intravenous, intraperitoneal, and / or subcutaneous administration. For injection, the compositions or components thereof of the present invention are formulated as sterile liquid solutions, preferably in physiologically compatible buffers or solutions, such as saline solutions, Hank solutions, or Ringer solutions. Alternatively, the compositions or components thereof can be formulated as solids and resuspended shortly before use. Lyophilized powder forms are also possible.

[0144] Administration can also be performed via mucosal, topical, or transdermal routes. For mucosal, topical, or transdermal administration, a penetrant suitable for the barrier to be penetrated is used in the formulation. Such penetrants are generally known in the art and include, for example, bile salts and clostridial acid derivatives for mucosal administration. Additionally, detergents can be used to facilitate penetration. Mucosal administration can also be achieved, for example, via nasal sprays or suppositories (rectally or vaginally).

[0145] For inhalation formulations, the compositions or components thereof of the present invention can be formulated as dry powders or suitable solutions, suspensions, or aerosols. Powders and solutions can be formulated with suitable additives known in the art. For example, powders may include suitable powder bases such as lactose or starch, and solutions may include propylene glycol, sterile water, ethanol, sodium chloride, and other additives such as acids, bases, and buffer salts. Such solutions or suspensions can be administered by inhalation via spray, pump, nebulizer, or nebulizer.

[0146] The topical formulations of the present invention can preferably be formulated as oils, creams, lotions, ointments, etc., by selecting suitable carriers known in the art. Emulsifiers, stabilizers, humectants, and antioxidants, as well as coloring or fragrance-importing agents, may also be included if desired. Topically applied creams are preferably formulated from a mixture of mineral oil, self-emulsifying beeswax, and water. Furthermore, transdermal application includes transdermal patches or dressings, such as bandages containing the active ingredient and optionally one or more carriers or diluents known in the art. For application via a transdermal delivery system, the dosage during the administration regimen will naturally be continuous, not intermittent.

[0147] The pharmaceutical compositions of the present invention can also be used in combination with other therapeutic agents for treating the same disease. Such combination includes administering, at different times, a composition comprising a direct or indirect supplement of oxidized nicotinamide adenine dinucleotide and / or other mitochondrial function-optimizing substances and a medicament for treating heart-related diseases, along with one or more other therapeutic agents, or a composition simultaneously comprising a direct or indirect supplement of oxidized nicotinamide adenine dinucleotide and / or other mitochondrial function-optimizing substances and a medicament for treating heart-related diseases, along with one or more other therapeutic agents. In some embodiments, the dosage of the compositions of the present invention comprising a direct or indirect supplement of oxidized nicotinamide adenine dinucleotide and / or other mitochondrial function-optimizing substances and a medicament for treating heart-related diseases, or other therapeutic agents used in combination, may be modified, for example, by reducing the dosage relative to the compound or therapeutic agent used alone using methods known to those skilled in the art.

[0148] Combined administration via any route of administration includes delivering the composition or components thereof of the present invention together with one or more other pharmaceutical therapeutic agents in any formulation via the same route of administration, including formulations in which two compounds are chemically linked and retain their respective therapeutic activities upon administration. In one aspect, other pharmaceutical therapies may be co-administered with the composition or components thereof of the present invention. Combined use by co-administration includes administering a formulation of co-formulation or chemically linked compounds, or administering two or more independent formulations of compounds over a short period (e.g., within one hour, two hours, three hours, up to 24 hours) via the same or different routes of administration.

[0149] In the description of this specification, references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0150] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0151] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A pharmaceutical composition for the prevention and / or treatment of diseases of the mammalian cardiac system, comprising a mitochondrial function-optimizing substance and a therapeutic agent for heart-related diseases.

2. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 1, characterized in that, The mitochondrial function-enhancing substance includes at least one of oxidized nicotinamide adenine dinucleotide and oxidized nicotinamide adenine dinucleotide supplement.

3. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 2, characterized in that, The oxidized nicotinamide adenine dinucleotide supplement includes nicotinamide adenine dinucleotide precursors.

4. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 3, characterized in that, The nicotinamide adenine dinucleotide precursor includes at least one of DL-β-(3-indolyl)-α-aminopropionic acid, pyridine-3-carboxylic acid, 3-pyridinecarboxamide, nicotinamide ribose, and β-nicotinamide mononucleotide.

5. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 1, characterized in that, The mitochondrial function-enhancing substances also include 3,3′,4′,7-tetrahydroxyflavone, 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid, Omega-3 fish oil, N,(3-aminopropyl)-1,4-diaminobutane, 3-N-acetyl-5-methoxytryptamine, N-[2-(2-iodo-5-methoxyindole-3-yl)ethyl]acetamide, N-[2-(6-fluoro-5-methoxy-1H-indole- At least one of the following: (3-yl)ethyl]acetamide, N-[2-(6-chloro-5-methoxy-1H-indol-3-yl)ethyl]acetamide, N-[2-(5-methoxy-2-phenyl-1H-indol-3-yl)ethyl]acetamide, (E)-5-[2-(4-hydroxyphenyl)-vinyl]-1,3-benzenediol, H2N-DArg-Dmt-Lys-Phe-NH2, ATP-promoting coenzymes, chlorophyll, B vitamins, curcumin, creatine, etc.

6. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to any one of claims 1 to 5, characterized in that, The medication for treating heart-related diseases includes a pharmaceutically active ingredient selected from at least one of the following: positive inotropic drugs, angiotensin II receptor antagonists, platelet aggregation inhibitors, lipid-lowering drugs, antiarrhythmic drugs, angiotensin-converting enzyme inhibitors, diuretics, arterial vasodilators, and 5-HT antagonists.

7. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 6, characterized in that, The positive inotropic drugs include phosphodiesterase inhibitors.

8. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 6, characterized in that, The positive inotropic drugs include at least one of pimoxendan, milrinone, amrinone, icariin, quinazoline, cilostazol, caffeine, aminophylline, 3-isobutyl-1-methylxanthine, paraxanthine, cyproterone acetate, theobromine, theophylline, vinpocetine, hydroxyindole, mesamidol, cyclolipan, isobutyllast, luteolin, drotavirin, piramylast, roflumilast, apralast, cetazidine, sildenafil, vardenafil, tadalafil, avanafil, udenafil, dipyridamole, papaverine, ammonia, cardiac glycosides, levosimendan, digitalis glycoside, digoxin, and strophanthidin K.

9. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 6, characterized in that, The angiotensin-converting enzyme inhibitors include at least one of captopril, cilazapril, quindopril, imidapril, benazepril, lisinopril, perindopril, quinapril, ramipril, enalapril, zolfenpril, and benazepril.

10. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 6, characterized in that, The angiotensin II receptor antagonist includes at least one of eprosartan, valsartan, irbesartan, flumasartan, azisartan, olmesartan, telmisartan, candesartan, losartan, and sacubitril / valsartan. And / or, The platelet aggregation inhibitors include at least one of cilostazol, aspirin, clopidogrel, trifluoromethanesulfonic acid, prasugrek, ticagrelor, troplotitin, tirofiban, aptifibatide, abciximab, vorapasal, tretroban, dipyridamole, ADP receptor antagonists, and GpIIb / IIIa receptor antagonists. And / or, The lipid-lowering drugs include at least one of simvastatin, atorvastatin, lovastatin, rosuvastatin, pravastatin, fluvastatin, pitavastatin, bezafibrate, probucol, acilimus, niacin, Src inhibitor Saracatinib, PCSK9 inhibitor Alirocumab, PCSK9 inhibitor Evolocumab, and myosin inhibitor Aficamten. And / or, The antiarrhythmic drugs include at least one of the following: digitalis, quinidine, procaine, disopyramide, phenytoin sodium, mexiletine, encaine, flecainide, propranolol, atenolol, diltiazem, amiodarone, metoprolol, propafenone, verapamil, lidocaine, ivabradine, bisoprolol, phentolamine, procainamide, ethoxymethazine, cefotaxime, acebutolol, and dicyclohexoperidine. And / or, The diuretic includes at least one of the following: amphotericin B, lithium citrate, tolvaptan, conivatan, dopamine, dazosulfan, acetazolamide, bumetanide, ethynyl acetylcholine, furosemide, torasemide, glucose, mannitol, eplerenone, spironolactone, amiloride, caffeine, theophylline, theobromine, benzylfluorothiazide, hydrochlorothiazide, cannic acid ester, and triamterene. And / or, The arterial vasodilators include at least one of adenosine, adrenaline, bradykinin, histamine, nicotinic acid, nitric oxide, diltiazem, verapamil, nifedipine, captopril, enalapril, hydralazine, pyrazinidine, acetylglycerol-ether-phosphocholine, prostacyclin, and isosorbide dinitrate. And / or, The 5-HT antagonist includes sargrelate, and at least one of fluoxetine, paroxetine, and sertraline.

11. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 6, characterized in that, The pharmaceutical composition comprises the nicotinamide adenine dinucleotide precursor, the positive inotropic drug, and / or the angiotensin-converting enzyme inhibitor and / or angiotensin receptor antagonist.

12. The pharmaceutical composition for the prevention and / or treatment of mammalian cardiac diseases according to claim 1, characterized in that, The pharmaceutical composition comprises at least one of oxidized nicotinamide adenine dinucleotide, 3,3′,4′,7-tetrahydroxyflavone, 4,5-dihydro-4,5-dioxo-1-hydropyrrolo(2,3-f)quinoline-2,7,9-tricarboxylic acid, (E)-5-[2-(4-hydroxyphenyl)-vinyl]-1,3-benzenediol, H2N-DArg-Dmt-Lys-Phe-NH2, nicotinamide ribose, β-nicotinamide mononucleotide, DL-β-(3-indolyl)-α-aminopropionic acid, pyridine-3-carboxylic acid, 3-pyridinecarboxamide, pimoxendan, benazepril, telmisartan, benazepril hydrochloride, hydrochlorothiazide, sargrelate hydrochloride, sacubitril / valsartan, quinapril hydrochloride, enalapril, azilsartan, and furosemide.

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