Method for increasing or improving nitric oxide signals or levels, and composition

Abstract

A method for increasing or improving nitric oxide signals or levels, the method comprising: administering a composition to a subject, the composition comprising: arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof. The method and the composition can be used to increase, improve, or enhance nitric oxide signals or levels, inhibit nitric oxide decomposition, and increase or enhance nitrous acid and / or cyclic guanosine monophosphate levels, and can improve fatigue resistance or enhance muscle strength, maintain or improve cardiovascular health, maintain or improve vascular function and structure, inhibit or mitigate endothelial cell damage, or mitigate insufficient blood flow.

Description

Methods and compositions for increasing or improving nitric oxide signal or level Technical Field

[0001] This invention belongs to the technical field of nutritional products or dietary supplements, and specifically relates to a method and composition for increasing or improving nitric oxide signal or level, and / or inhibiting nitric oxide decomposition, and / or increasing or improving the levels of nitrite and / or cyclic guanosine monophosphate. Background Technology

[0002] Nitric oxide (NO), also known as nitrogen oxides, is produced by almost every cell in the human body. It is considered a mediator of intercellular communication and plays a vital role in numerous bodily processes, including inflammation, vasodilation, and neurotransmission. First and foremost, it is considered one of the most important molecules when it comes to our vascular health. For instance, NO plays a crucial role in vasodilation and can also lower blood pressure. NO is a signaling molecule widely present in the cardiovascular system and various cell types, and it prevents the occurrence and development of atherosclerosis in multiple ways; simply put, NO can improve endothelial function. NO also has the function of scavenging oxygen free radicals, including preventing the oxidation of low-density lipoprotein cholesterol. NO can regulate the central nervous system and improve neurodegeneration. NO can also improve tolerance to both aerobic and anaerobic exercise. NO plays an important regulatory role in glucose uptake by skeletal muscle during contraction / exercise. Therefore, enhancing NO signaling is particularly important.

[0003] Currently, products on the market that enhance NO signaling can cause side effects such as nausea, vomiting, and palpitations. Therefore, it is crucial to find a product that can enhance NO signaling without side effects. Summary of the Invention

[0004] One aspect of the present invention relates to a method for increasing or improving nitric oxide signaling or levels, the method comprising: administering to a subject a composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

[0005] In some implementations, the method increases or enhances the levels of nitrite and / or cyclic guanosine monophosphate.

[0006] In some implementations, the method improves fatigue resistance or enhances muscle strength, maintains or improves cardiovascular health, maintains or improves vascular function and structure, inhibits or reduces endothelial cell damage, or reduces insufficient blood flow.

[0007] In some embodiments, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 10%, 20%, 30%, 40%, 50%, 55%, or 60% compared to subjects who did not receive the composition.

[0008] In some embodiments, compared with subjects who received arginine alone or a pharmaceutically acceptable salt, ester, or derivative thereof, the nitric oxide signal or level of the subject receiving the composition increased or improved by more than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%. In some embodiments, the dosage of the composition is 0.1-20 g or 0.1 mM to 1 M or 0.001-90% (w / w).

[0009] In some embodiments, the composition is administered by a single dose or multiple fractions, with the dosage and / or daily dose being 0.2-18 g, 0.3-16 g, 0.5-14 g, 0.8-12 g, 1-10 g, or 2-8 g. In some embodiments, the dosage and / or daily dose of the composition is 0.2 mM to 10 mM, 0.5 mM to 50 mM, 1 mM to 100 mM, 2 mM to 500 mM, 2 mM to 100 mM, 5 mM to 200 mM, 5 mM to 500 mM, 10 mM to 500 mM, or 10 mM to 1 M. In some embodiments, the dosage and / or daily dose of the composition is 0.001-50%, 0.005-85%, 0.01-80%, 0.05-75%, 0.5-70%, 1-50%, 2-45%, 50-95%, 40-85%, 30-80%, 20-75%, 10-70%, or 5-65% (w / w).

[0010] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:1.

[0011] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:25 to 250:1, 1:20 to 200:1, 1:15 to 150:1, 1:10 to 125:1, 1:10 to 50:1, 1:10 to 10:1, 1:8 to 100:1, 1:5 to 80:1, 1:3 to 50:1, 1:2 to 30:1, or 1:1 to 10:1.

[0012] In some implementations, the subjects are humans or mammals.

[0013] In some embodiments, the composition is formulated as a nutritional product, dietary supplement, food, beverage, or animal feed.

[0014] In some embodiments, the composition is in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, or syrups.

[0015] In some embodiments, the composition further comprises a component that enhances nitric oxide signaling or levels, such component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

[0016] In some implementations, administration is carried out via a route selected from: oral, intravenous, intramuscular, intraperitoneal, or sublingual administration.

[0017] Another aspect of the invention relates to a composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof, the composition being used to increase or improve nitric oxide signaling or levels.

[0018] In some embodiments, the composition increases or enhances the levels of nitrite and / or cyclic guanosine monophosphate.

[0019] In some embodiments, the composition enhances anti-fatigue capabilities or strengthens muscle strength, maintains or improves cardiovascular health, maintains or improves vascular function and structure, inhibits or reduces endothelial cell damage, or alleviates insufficient blood flow.

[0020] In some embodiments, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 10%, 20%, 30%, 40%, 50%, 55%, or 60% compared to subjects who did not receive the composition.

[0021] In some implementations, compared with subjects who received arginine alone or a pharmaceutically acceptable salt, ester, or derivative thereof, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

[0022] In some embodiments, the composition is applied in amounts of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

[0023] In some embodiments, the composition is administered by a single dose or multiple fractions, with the dosage and / or daily dose being 0.2-18 g, 0.3-16 g, 0.5-14 g, 0.8-12 g, 1-10 g, or 2-8 g. In some embodiments, the dosage and / or daily dose of the composition is 0.2 mM to 10 mM, 0.5 mM to 50 mM, 1 mM to 100 mM, 2 mM to 500 mM, 2 mM to 100 mM, 5 mM to 200 mM, 5 mM to 500 mM, 10 mM to 500 mM, or 10 mM to 1 M. In some embodiments, the dosage and / or daily dose of the composition is 0.001-50%, 0.005-85%, 0.01-80%, 0.05-75%, 0.5-70%, 1-50%, 2-45%, 50-95%, 40-85%, 30-80%, 20-75%, 10-70%, or 5-65% (w / w).

[0024] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:1.

[0025] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:25 to 250:1, 1:20 to 200:1, 1:15 to 150:1, 1:10 to 125:1, 1:10 to 50:1, 1:10 to 10:1, 1:8 to 100:1, 1:5 to 80:1, 1:3 to 50:1, 1:2 to 30:1, or 1:1 to 10:1.

[0026] In some embodiments, the composition is formulated as a nutritional product, dietary supplement, food, beverage, or animal feed.

[0027] In some embodiments, the composition is in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, or syrups.

[0028] In some embodiments, the composition further comprises a component that enhances nitric oxide signaling or levels, such component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

[0029] Another aspect of the invention relates to the use of a composition in the preparation of a nutrient, dietary supplement, food, beverage, or animal feed for increasing or improving nitric oxide signaling or levels, the composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

[0030] In some embodiments, the composition increases or enhances the levels of nitrite and / or cyclic guanosine monophosphate.

[0031] In some embodiments, the composition enhances anti-fatigue capabilities or strengthens muscle strength, maintains or improves cardiovascular health, maintains or improves vascular function and structure, inhibits or reduces endothelial cell damage, or alleviates insufficient blood flow.

[0032] In some embodiments, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 10%, 20%, 30%, 40%, 50%, 55%, or 60% compared to subjects who did not receive the composition.

[0033] In some implementations, compared with subjects who received arginine alone or a pharmaceutically acceptable salt, ester, or derivative thereof, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

[0034] In some embodiments, the composition is applied in amounts of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

[0035] In some embodiments, the composition is administered by a single dose or multiple fractions, with the dosage and / or daily dose being 0.2-18 g, 0.3-16 g, 0.5-14 g, 0.8-12 g, 1-10 g, or 2-8 g. In some embodiments, the dosage and / or daily dose of the composition is 0.2 mM to 10 mM, 0.5 mM to 50 mM, 1 mM to 100 mM, 2 mM to 500 mM, 2 mM to 100 mM, 5 mM to 200 mM, 5 mM to 500 mM, 10 mM to 500 mM, or 10 mM to 1 M. In some embodiments, the dosage and / or daily dose of the composition is 0.001-50%, 0.005-85%, 0.01-80%, 0.05-75%, 0.5-70%, 1-50%, 2-45%, 50-95%, 40-85%, 30-80%, 20-75%, 10-70%, or 5-65% (w / w).

[0036] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:1.

[0037] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:25 to 250:1, 1:20 to 200:1, 1:15 to 150:1, 1:10 to 125:1, 1:10 to 50:1, 1:10 to 10:1, 1:8 to 100:1, 1:5 to 80:1, 1:3 to 50:1, 1:2 to 30:1, or 1:1 to 10:1.

[0038] In some embodiments, the composition is in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, or syrups.

[0039] In some embodiments, the composition further comprises a component that enhances nitric oxide signaling or levels, such component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

[0040] Another aspect of the present invention relates to a method for increasing or raising the levels of nitrite and / or cyclic guanosine monophosphate, comprising: administering to a subject a composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

[0041] In some implementations, the method improves fatigue resistance or enhances muscle strength, maintains or improves cardiovascular health, maintains or improves vascular function and structure, inhibits or reduces endothelial cell damage, or reduces insufficient blood flow.

[0042] In some embodiments, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 10%, 20%, 30%, 40%, 50%, 55%, or 60% compared to subjects who did not receive the composition.

[0043] In some implementations, compared with subjects who received arginine alone or a pharmaceutically acceptable salt, ester, or derivative thereof, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

[0044] In some embodiments, the composition is applied in amounts of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

[0045] In some embodiments, the composition is administered by a single dose or multiple fractions, with the dosage and / or daily dose being 0.2-18 g, 0.3-16 g, 0.5-14 g, 0.8-12 g, 1-10 g, or 2-8 g. In some embodiments, the dosage and / or daily dose of the composition is 0.2 mM to 10 mM, 0.5 mM to 50 mM, 1 mM to 100 mM, 2 mM to 500 mM, 2 mM to 100 mM, 5 mM to 200 mM, 5 mM to 500 mM, 10 mM to 500 mM, or 10 mM to 1 M. In some embodiments, the dosage and / or daily dose of the composition is 0.001-50%, 0.005-85%, 0.01-80%, 0.05-75%, 0.5-70%, 1-50%, 2-45%, 50-95%, 40-85%, 30-80%, 20-75%, 10-70%, or 5-65% (w / w).

[0046] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:1.

[0047] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:25 to 250:1, 1:20 to 200:1, 1:15 to 150:1, 1:10 to 125:1, 1:10 to 50:1, 1:10 to 10:1, 1:8 to 100:1, 1:5 to 80:1, 1:3 to 50:1, 1:2 to 30:1, or 1:1 to 10:1.

[0048] In some implementations, the subjects are humans or mammals.

[0049] In some embodiments, the composition is formulated as a nutritional product, dietary supplement, food, beverage, or animal feed.

[0050] In some embodiments, the composition is in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, or syrups.

[0051] In some embodiments, the composition further comprises a component that enhances nitric oxide signaling or levels, such component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

[0052] In some implementations, administration is carried out via a route selected from: oral, intravenous, intramuscular, intraperitoneal, or sublingual administration.

[0053] Another aspect of the invention relates to the use of a composition in the preparation of a nutritional product, dietary supplement, food, beverage, or animal feed for increasing or enhancing the levels of nitrite and / or cyclic guanosine monophosphate, the composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

[0054] In some implementations, the method improves fatigue resistance or enhances muscle strength, maintains or improves cardiovascular health, maintains or improves vascular function and structure, inhibits or reduces endothelial cell damage, or reduces insufficient blood flow.

[0055] In some embodiments, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 10%, 20%, 30%, 40%, 50%, 55%, or 60% compared to subjects who did not receive the composition.

[0056] In some implementations, compared with subjects who received arginine alone or a pharmaceutically acceptable salt, ester, or derivative thereof, the nitric oxide signal or level of subjects who received the composition increased or improved by more than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

[0057] In some embodiments, the composition is applied in amounts of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

[0058] In some embodiments, the composition is administered by a single dose or multiple fractions, with the dosage and / or daily dose being 0.2-18 g, 0.3-16 g, 0.5-14 g, 0.8-12 g, 1-10 g, or 2-8 g. In some embodiments, the dosage and / or daily dose of the composition is 0.2 mM to 10 mM, 0.5 mM to 50 mM, 1 mM to 100 mM, 2 mM to 500 mM, 2 mM to 100 mM, 5 mM to 200 mM, 5 mM to 500 mM, 10 mM to 500 mM, or 10 mM to 1 M. In some embodiments, the dosage and / or daily dose of the composition is 0.001-50%, 0.005-85%, 0.01-80%, 0.05-75%, 0.5-70%, 1-50%, 2-45%, 50-95%, 40-85%, 30-80%, 20-75%, 10-70%, or 5-65% (w / w).

[0059] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:1.

[0060] In some embodiments, the ratio of arginine or its pharmaceutically acceptable salts, esters, or derivatives to tetrahydrocurcumin and / or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:25 to 250:1, 1:20 to 200:1, 1:15 to 150:1, 1:10 to 125:1, 1:10 to 50:1, 1:10 to 10:1, 1:8 to 100:1, 1:5 to 80:1, 1:3 to 50:1, 1:2 to 30:1, or 1:1 to 10:1.

[0061] In some embodiments, the composition is in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, or syrups.

[0062] In some embodiments, the composition further comprises a component that enhances nitric oxide signaling or levels, such component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

[0063] The present invention relates to compositions comprising arginine or pharmaceutically acceptable salts, esters, or derivatives thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or pharmaceutically acceptable salts or esters thereof, and their administration thereof, which can be used to increase or improve or enhance nitric oxide signaling or levels, inhibit nitric oxide decomposition, and increase or enhance nitrite and / or cyclic guanosine monophosphate levels, improve anti-fatigue capabilities or enhance muscle strength, maintain or improve cardiovascular health, maintain or improve vascular function and structure, inhibit or reduce endothelial cell damage, or alleviate insufficient blood flow. These compositions play a vital role in the normal function of the cardiovascular, nervous, pulmonary, gastrointestinal, renal, and immune systems. Attached Figure Description

[0064] Figure 1 shows the cell activity of different doses (1 mM, 10 mM) of Arg.

[0065] Figure 2 shows the cell activity of different doses (1 mM, 10 mM) of 4HC and BAIBA.

[0066] Figure 3 illustrates the role of Arg and Arg+4HC in promoting NO secretion.

[0067] Figure 4 illustrates the role of Arg and Arg + BAIBA in promoting NO secretion. Detailed Implementation

[0068] The present invention will now be further described with reference to preferred embodiments thereof. While the invention will be described in conjunction with preferred embodiments, it should be understood that they are not intended to limit the invention to these embodiments. Rather, the invention is intended to cover alternatives, modifications, and equivalents that may be included within the spirit and scope of the invention as defined in the claims.

[0069] As used herein, the term “or” is intended to include both “and” and “or”. In other words, the term “or” can also be replaced with “and / or”.

[0070] As used herein, unless the context clearly indicates otherwise, the singular forms “a / an” and “the” are intended to include the plural forms as well.

[0071] As used herein, the terms “comprising” or “including” or variations thereof refer to items that are included in the context of the term in their non-restrictive sense, but do not exclude items not specifically mentioned. It also includes the more restrictive verbs 'consistently composed of' and 'comprises of'.

[0072] As used herein, the terms “subject” and “individual” are used interchangeably to refer to any subject to whom the methods and compositions of this disclosure may be applied or administered. A subject may have a disease or ailment, but does not need to be ill to benefit from the methods and compositions of this disclosure. Any subject may take the disclosed compositions or become a recipient of the disclosed methods. In this document, the term “subject” refers to an animal (e.g., birds, reptiles, and mammals). In some embodiments, the subject may be a mammal including non-primates (e.g., camels, donkeys, zebras, cows, horses, cats, dogs, rats, and mice) and primates (e.g., monkeys, chimpanzees, and humans). In some embodiments, the subject may be a non-human mammal. In other embodiments, the subject may be a human.

[0073] As used herein, the term "administration" refers to the process of delivering the disclosed composition or active ingredient to a subject. The compositions of the present invention are preferably administered via oral, intravenous, intramuscular, intraperitoneal, or sublingual routes, but may also be administered via other conventional routes to achieve the desired effect.

[0074] As used herein, the term “pharmaceutical acceptable” means pharmaceutically, physiologically, or dietaryly acceptable, and refers to combinations of compositions or reagents, materials or compositions and / or dosage forms thereof that are suitable for contact with human and animal tissues, compatible with other components of the composition, without excessive toxicity, irritation, allergic reactions or other problems or complications, and commensurate with a reasonable benefit / risk ratio, within the bounds of reasonable medical judgment.

[0075] Improving fatigue resistance or enhancing muscle strength refers to promoting the body's synthesis of nitric oxide. Nitric oxide effectively promotes blood circulation and increases blood flow, thereby carrying more oxygen and nutrients to the target muscle groups, enhancing the muscle pump, and improving training effectiveness. At the same time, nitric oxide inhibits the production of lactic acid and urea nitrogen during exercise, reducing muscle soreness and having an anti-muscle fatigue effect.

[0076] Maintaining or improving cardiovascular health means that nitric oxide can reduce the risk of cardiovascular diseases such as heart disease and stroke by improving blood circulation and lowering blood pressure. Furthermore, nitric oxide is also crucial for maintaining the health of the vascular endothelium, helping to prevent diseases such as atherosclerosis.

[0077] Maintaining or improving vascular function and structure means that nitric oxide can relax the smooth muscle in blood vessels, making the blood vessels larger. As a result, the blood flow in the body can be greater, and more oxygen and nutrients can be carried.

[0078] Inhibiting or mitigating endothelial cell damage refers to increasing nitric oxide production by increasing the activity of nitric oxide synthase in endothelial cells, and also to enhancing the activity of superoxide dismutase in cells, thereby reducing oxidative damage to endothelial cells caused by oxidized low-density lipoprotein.

[0079] Nitric oxide can alleviate insufficient blood flow because it relaxes vascular smooth muscle, increases blood circulation, and thus relieves insufficient blood supply.

[0080] The present invention comprises arginine or its pharmaceutically acceptable salts, esters, or derivatives; and tetrahydrocurcumin and / or β-aminoisobutyric acid or their pharmaceutically acceptable salts or esters, which can be used with dietaryly or pharmaceutically acceptable carriers. The aforementioned carriers include non-toxic compatible substances commonly used in health foods, dietary supplements, and pharmaceutical preparations, such as sugars, starches, cellulose and their derivatives, powdered tragacanth gum, malt, gelatin, talc, oils, glycols, polyols, esters, agar, alginic acid, pyrogen-free water, isotonic saline, etc.

[0081] In some embodiments, the compositions of the present invention comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof may be administered with other supplements, such as vitamins, minerals, nootropics, and other supplements known in the art.

[0082] The compositions of the present invention and various formulations thereof are considered; the compositions will be formulated as nutritional supplements or dietary supplements, (medical) foods, beverages, animal feeds, in liquid or solid form. For oral administration, the compositions of the present invention may be in any suitable form, including solutions, tablets, gel capsules, capsules, or alternative nutritional foods or supplements.

[0083] The intended method involves administering daily an appropriate amount of a combination containing arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof, depending on the specific formulation and form. The amount to be administered may also vary depending on factors such as the subject's sensitivity, age, sex, weight, and specific response. One or more doses may be administered once or multiple times daily at any time period or at a suitable frequency. For example, an effective dose may be administered daily for one day, several days, multiple days, or indefinitely.

[0084] Unless otherwise specified, the materials and reagents described in the following examples are all commercially available products that can be purchased from the market. Example Nitric Oxide (NO) Secretion Promotion Assay – This assay tests the effect of the composition of the present invention on promoting NO secretion in human umbilical vein endothelial cells.

[0085] The reagents used included: endothelial cell basal culture medium, fetal bovine serum (FBS), antibiotics, endothelial cell culture additives, CCK-8 cell viability assay kit, serum-free cryopreservation solution, 0.25% trypsin, NO assay kit, Arg (arginine), 4HC (tetrahydrocurcumin), and BAIBA (β-aminoisobutyric acid). The instruments used included: cell culture incubator, cell counter, and microplate reader.

[0086] Cell Culture: Human umbilical vein endothelial cells (HUVECs) were purchased from Wuhan Saiss Biotechnology Co., Ltd. Cells were maintained in a humid environment (95% air and 5% carbon dioxide, 37°C) and cultured in endothelial cell basal medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 1% endothelial cell culture additive, and 1% penicillin / streptomycin. Cultures were harvested, and cell morphology was examined under a microscope. Cell counts were performed on the cell suspension using a hemocytometer (Marienfeld, Germany) to monitor cell number.

[0087] Cytotoxicity assay: The effect of the composition of the present invention on the viability of normal human umbilical vein endothelial cells was evaluated using the CCK-8 assay. Briefly, cells were seeded in 96-well plates and treated with 1 mM and 10 mM of the composition of the present invention for 24 h, followed by incubation with CCK-8 solution (100 μL / well) for 2 h. Absorbance at 450 nm was measured using a microplate reader.

[0088] Reagent Preparation: Arg Test Solution Preparation: Weigh 7.0 mg of Arg, dissolve in 4 mL of complete endothelial cell culture medium, and filter through a 0.22 μm filter to obtain a 10 mM sample solution. 4HC Test Solution Preparation: Weigh 14.8 mg of 4HC, dissolve in 4 mL of complete endothelial cell culture medium, and filter through a 0.22 μm filter to obtain a 10 mM sample solution. BAIBA Test Solution Preparation: Weigh 4.2 mg of BAIBA, dissolve in 4 mL of complete endothelial cell culture medium, and filter through a 0.22 μm filter to obtain a 10 mM sample solution. Arg + 4HC Test Solution Preparation: Mix 10 mM Arg solution and 10 mM 4HC solution 1:1 thoroughly. Arg + BAIBA Test Solution Preparation: Mix 10 mM Arg solution and 10 mM 4HC solution 1:1 thoroughly.

[0089] Composition to stimulate NO secretion: HUVECs were seeded in 24-well plates (1×10⁻⁶). 4Cells were cultured at ~80% confluence (cells / mL). The supernatant was discarded, and the cells were washed once with phosphate-buffered saline (PBS) at room temperature. Endothelial cell basal culture medium was added to the cells and incubated in an incubator for 1 h. The basal culture medium was then replaced with medium containing Arg (non-cytotoxic dose, 10 mM), 4HC (non-cytotoxic dose, 10 mM), Arg + 4HC (non-cytotoxic dose, 5 mM + 5 mM), BAIBA (non-cytotoxic dose, 10 mM), Arg + BAIBA (non-cytotoxic dose, 5 mM + 5 mM), or a control (DMSO). The cells were then returned to the incubator and incubated for 24 h. The cell culture supernatant was collected and used after overnight incubation at 4°C.

[0090] NO detection: The NO content is directly detected using a kit. NO is readily oxidized to NO2- in an aqueous solution in vivo. Under acidic conditions, NO2- reacts with diazonium sulfonate amine to form a diazonium compound, which further couples with naphthylvinyldiamine. The product has a characteristic peak at 550 nm. The absorbance is measured to calculate the NO content.

[0091] As shown in the table below, the sample solution, reagent 1, and reagent 2 were added sequentially to the 96-well plate, mixed well, and allowed to stand at room temperature for 15 minutes. The absorbance was then measured at 550 nm.

[0092]

[0093] The absorbance value of the blank tube is A (blank), and the absorbance value of the test tube is A (test). The NO content is calculated using the following formula: NO content (μmol / L) = (△A + 0.0103) ÷ 0.008 × Vtotalreaction ÷ Vsample = 250 × (△A + 0.0103), where Vtotalreaction: total reaction volume, 0.2 mL; Vsample: sample volume in the reaction, 0.1 mL; Vtotalsample: volume of extract added, 1 mL.

[0094] Collect and analyze the data.

[0095] Figures 1 and 2 show the cell viability of different doses (1 mM, 10 mM) of Arg, 4HC, and BAIBA, respectively. As shown in Figures 1 and 2, all doses of the substances were non-toxic to the cells, and cell viability was approximately 100%. * indicates a statistically significant difference compared to the control group, and ** indicates p < 0.01.

[0096] Figure 3 illustrates the effects of Arg, 4HC, and Arg + 4HC on promoting NO secretion in umbilical vein endothelial cells. As shown in Figure 3, combined use with 4HC promotes arginine NO production. The NO secretion in the Arg + 4HC group was 29% higher than that of 4HC alone, 26% higher than that of Arg alone, and approximately 37% higher than the control group. The Arg group showed approximately 16% higher NO secretion than the control group, and the 4HC group showed approximately 11% higher NO secretion than the control group. # indicates a statistically significant difference compared to the control group, * indicates a statistically significant difference between the two groups, ## indicates p < 0.01, ### indicates p < 0.001, and ** indicates p < 0.01.

[0097] Figure 4 illustrates the effects of Arg, BAIBA, and Arg + BAIBA on promoting NO secretion in umbilical vein endothelial cells. As shown in Figure 4, combined use with BAIBA promotes the production of arginine NO. The NO secretion in the Arg + BAIBA combination group was 23% higher than that of BAIBA alone, 18% higher than that of Arg alone, and 32% higher than that of the control group. The Arg group was approximately 16% higher than the control group, and the BAIBA group was approximately 12% higher than the control group. # indicates a statistically significant difference compared to the control group, * indicates a statistically significant difference between the two groups, ## indicates p < 0.01, ** indicates p < 0.01, and *** indicates p < 0.001.

[0098] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any person skilled in the art can make various changes, modifications, substitutions and variations to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the claims and their equivalents.

Claims

1. A method for increasing or improving nitric oxide signal or level, characterized in that, The method includes administering a composition to a subject, the composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

2. The method according to claim 1, characterized in that, The method increases or enhances the levels of nitrite and / or cyclic guanosine monophosphate.

3. The method according to claim 1 or 2, characterized in that, The methods described improve fatigue resistance or enhance muscle strength, maintain or improve cardiovascular health, maintain or improve vascular function and structure, inhibit or reduce endothelial cell damage, or reduce insufficient blood flow.

4. The method according to any one of claims 1-3, characterized in that, Compared with subjects who received arginine alone or its pharmaceutically acceptable salts, esters, or derivatives, subjects who received the composition showed an increase or improvement in nitric oxide signal or level of greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

5. The method according to any one of claims 1-4, characterized in that, The composition is applied at a rate of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

6. The method according to any one of claims 1-5, characterized in that, The ratio of the arginine or its pharmaceutically acceptable salts, esters, or derivatives to the tetrahydrocurcumin or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:

1.

7. The method according to any one of claims 1-6, characterized in that, The subjects are humans or mammals.

8. The method according to any one of claims 1-7, characterized in that, The composition is formulated into nutritional products, dietary supplements, food, beverages, or animal feed.

9. The method according to any one of claims 1-8, characterized in that, The composition is available in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, and syrups.

10. The method according to any one of claims 1-9, characterized in that, The composition further comprises a component that enhances nitric oxide signaling or levels, said component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

11. The method according to any one of claims 1-10, characterized in that, The administration is carried out via a route selected from the following: oral, intravenous, intramuscular, intraperitoneal, or sublingual administration.

12. A composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof, characterized in that, The composition is used to increase or improve nitric oxide signal or level.

13. The composition according to claim 12, characterized in that, The composition increases or enhances the levels of nitrite and / or cyclic guanosine monophosphate.

14. The composition according to claim 12 or 13, characterized in that, The composition enhances anti-fatigue ability or strengthens muscle strength, maintains or improves cardiovascular health, maintains or improves vascular function and structure, inhibits or reduces endothelial cell damage, or reduces insufficient blood flow.

15. The composition according to any one of claims 12-14, characterized in that, Compared with subjects who received arginine alone or its pharmaceutically acceptable salts, esters, or derivatives, subjects who received the composition showed an increase or improvement in nitric oxide signal or level of greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

16. The composition according to any one of claims 12-15, characterized in that, The composition is applied at a rate of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

17. The composition according to any one of claims 12-16, characterized in that, The ratio of the arginine or its pharmaceutically acceptable salts, esters, or derivatives to the tetrahydrocurcumin or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:

1.

18. The composition according to any one of claims 12-17, characterized in that, The composition is formulated into nutritional products, dietary supplements, food, beverages, or animal feed.

19. The composition according to any one of claims 12-18, characterized in that, The composition is available in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, and syrups.

20. The composition according to any one of claims 12-19, characterized in that, The composition further comprises a component that enhances nitric oxide signaling or levels, said component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

21. Use of a composition in the preparation of a nutrient, dietary supplement, food, beverage, or animal feed for increasing or improving nitric oxide signaling or levels, characterized in that, The composition comprises arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

22. The use according to claim 21, characterized in that, The composition increases or enhances the levels of nitrite and / or cyclic guanosine monophosphate.

23. The use according to claim 21 or 22, characterized in that, The composition enhances anti-fatigue ability or strengthens muscle strength, maintains or improves cardiovascular health, maintains or improves vascular function and structure, inhibits or reduces endothelial cell damage, or reduces insufficient blood flow.

24. The use according to any one of claims 21-23, characterized in that, Compared with subjects who received arginine alone or its pharmaceutically acceptable salts, esters, or derivatives, subjects who received the composition showed an increase or improvement in nitric oxide signal or level of greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

25. The use according to any one of claims 21-24, characterized in that, The composition is applied at a rate of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

26. The use according to any one of claims 21-25, characterized in that, The ratio of the arginine or its pharmaceutically acceptable salts, esters, or derivatives to the tetrahydrocurcumin or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:

1.

27. The use according to any one of claims 21-26, characterized in that, The composition is available in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, and syrups.

28. The use according to any one of claims 21-27, characterized in that, The composition further comprises a component that enhances nitric oxide signaling or levels, said component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

29. A method for increasing or raising the levels of nitrite and / or cyclic guanosine monophosphate, characterized in that, The method includes administering a composition to a subject, the composition comprising arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

30. The method according to claim 29, characterized in that, The methods described improve fatigue resistance or enhance muscle strength, maintain or improve cardiovascular health, maintain or improve vascular function and structure, inhibit or reduce endothelial cell damage, or reduce insufficient blood flow.

31. The method according to claim 29 or 30, characterized in that, Compared with subjects who received arginine alone or its pharmaceutically acceptable salts, esters, or derivatives, subjects who received the composition showed an increase or improvement in nitrite and / or cyclic guanosine monophosphate levels of greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

32. The method according to any one of claims 29-31, characterized in that, The composition is applied at a rate of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

33. The method according to any one of claims 29-32, characterized in that, The ratio of the arginine or its pharmaceutically acceptable salts, esters, or derivatives to the tetrahydrocurcumin or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:

1.

34. The method according to any one of claims 29-33, characterized in that, The subjects are humans or mammals.

35. The method according to any one of claims 29-34, characterized in that, The composition is formulated into nutritional products, dietary supplements, food, beverages, or animal feed.

36. The method according to any one of claims 29-35, characterized in that, The composition is available in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, and syrups.

37. The method according to any one of claims 29-36, characterized in that, The composition further comprises a component that enhances nitric oxide signaling or levels, said component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

38. The method according to any one of claims 29-37, characterized in that, The administration is carried out via a route selected from the following: oral, intravenous, intramuscular, intraperitoneal, or sublingual administration.

39. Use of a composition in the preparation of nutritional products, dietary supplements, foods, beverages, or animal feeds for increasing or enhancing levels of nitrite and / or cyclic guanosine monophosphate, characterized in that, The composition comprises arginine or a pharmaceutically acceptable salt, ester, or derivative thereof; and tetrahydrocurcumin and / or β-aminoisobutyric acid or a pharmaceutically acceptable salt or ester thereof.

40. The use according to claim 39, characterized in that, The methods described improve fatigue resistance or enhance muscle strength, maintain or improve cardiovascular health, maintain or improve vascular function and structure, inhibit or reduce endothelial cell damage, or reduce insufficient blood flow.

41. The use according to claim 39 or 40, characterized in that, Compared with subjects who received arginine alone or its pharmaceutically acceptable salts, esters, or derivatives, subjects who received the composition showed an increase or improvement in nitrite and / or cyclic guanosine monophosphate levels of greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

42. The use according to any one of claims 39-41, characterized in that, The composition is applied at a rate of 0.1-20 g, or 0.1 mM to 1 M, or 0.001-90% (w / w).

43. The use according to any one of claims 39-42, characterized in that, The ratio of the arginine or its pharmaceutically acceptable salts, esters, or derivatives to the tetrahydrocurcumin or β-aminoisobutyric acid or its pharmaceutically acceptable salts or esters is 1:30 to 300:

1.

44. The use according to any one of claims 39-43, characterized in that, The composition is available in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, liniments, tinctures, tonics, liquid suspensions, and syrups.

45. The use according to any one of claims 39-44, characterized in that, The composition further comprises a component that enhances nitric oxide signaling or levels, said component being coenzyme Q10, vitamin E, vitamin C, carnosine, lycopene, creatine, cysteine, resveratrol, taurine, ergothioneine, glutathione, and / or citrulline and its salts.

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