Botanical composition for prolonged duration of muscle pump

Abstract

The invention provides nutraceutical compositions for modulating the nitric oxide synthase (NOS) pathways comprising mango fruit powder; black ginger extract; pine bark extract; and ginkgo biloba extract powder. The modulation of the NOS pathway with the nutraceutical compositions supports anti-inflammatory activities and vasodilation to enhance muscle activity.

Description

BOTANICAL COMPOSITION FOR PROLONGED DURATION OF MUSCLE PUMPREFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit to and priority to US Provisional Patent Application Serial No. 63 / 634,269, filed on April 15, 2024, which is hereby incorporated by this reference in its entirety.FIELD

[0002] This invention relates generally to nutraceutical compositions for significantly prolonging the duration of a muscle pump by supporting enhanced vasodilation, blood flow, and antioxidant levels and methods of making the same.BACKGROUND

[0003] A goal of many athletes and other body-conscious individuals is to maximize workouts and attain peak physical condition. Although steroids and other drugs have been used for this purpose, they have been associated with potentially harmful side effects. As a result, use of steroids and other drugs is disfavored and, in most instances, is even considered illegal.

[0004] Within especially the bodybuilding community, the ‘'pump” or “muscle pump” are terms that describes a feeling bodybuilders may experience when working out. This feeling, which results from blood engorging the muscles being worked, maybe experienced during a workout, as well as for some period after the completion of a workout. The more strenuous the workout, the greater the resulting “pump” would be expected. The “pump” is perceived by some as leading to larger increases in muscle mass and strength from a workout. Thus, increased flow- of blood through the veins may be desired in connection with a workout.

[0005] Muscle pump is associated with an increased blood flow to the muscles, resulting in improved nutrient delivery, waste removal, and a visually enhanced appearance. Traditional supplements focus on the temporary- induction of this state. However, there remains a significant need for a formulation capable of not only- inducing but also extending the duration of a muscle pump to maximize these benefits over time.

[0006] Provided herein are nutraceutical compositions significantly prolonging the duration of a muscle pump by supporting enhanced vasodilation, blood flow-, and antioxidant levels and methods of making the same. These nutraceutical compositionsuniquely include components that support anti-inflammatory activities, notably through the modulation of nitric oxide synthase (NOS) pathways, thereby promoting an optimal environment for sustained muscle pump and performance enhancement., and methods of making the same.SUMMARY

[0007] In accordance with the purpose(s) of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to a nutraceutical composition for modulating the nitric oxide synthase (NOS) pathways comprising: mango fruit powder; black ginger extract; pine bark extract; and ginkgo biloba extract powder; wherein modulation of the NOS pathway supports anti-inflammatory activities and vasodilation to enhance muscle activity.

[0008] In one embodiment, the mango fruit powder is a powder of a Mangifera indica L fruit. In another embodiment, the black ginger extract is extracted from a Kaempferia parviflora root. In yet another embodiment, the pine bark extract is extracted from a Pinus massoniana Lamb or aPinus pinaster bark. In one other embodiment, the ginkgo biloba extract powder is a powder of a dried whole leaf of Ginkgo biloba.

[0009] Another embodiment provides that the nutraceutical composition is comprised of 32-57% mango fruit powder, 23-36% black ginger extract, 13-25% pine bark extract, and 10-21% gingko biloba extract powder.

[0010] In another aspect, the invention relates to a method of modulating the nitric oxide synthase (NOS) pathways of a subject comprising administering to the subject the nutraceutical composition described above, wherein modulating the NOS pathways enhances a subject’s physical performance by prolonging the duration of a muscle pump.

[0011] In one embodiment, modulating the nitric oxide synthase (NOS) pathways enhances the subject’s vasodilation, blood flow, and antioxidant levels. In an additional embodiment, modulating the nitric oxide synthase (NOS) pathways supports improved nutrient delivery and metabolic waste removal over an extended period.

[0012] Additional 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. The advantages of the invention will be realizedand attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.BRIEF DESCRIPTIONS OF THE DRAWINGS

[0013] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate (one) several embodiment(s) of the invention and together with the description, serve to explain the principles of the invention.

[0014] Figure 1 shows the process for making the nutraceutical compositions. Included are processes for Ginko Powder, Mangifera Indica, Black Ginger Root, and Dry Pine Bark. The Ginko Powder is cleaned, and then undergoes an ethanol extraction, after which it is concentrated and spray dried. After sifting and mixing it is analyzed and added to the final blend. Mangifera Indica is cleaned and subjected to pulping, and extraction before being mixed with carriers. After mixing, it is blended, spray dried, sieved, and analyzed before being added to the final blend. Black Ginger root is cleaned, cut into slices, dried, and pulverized before extraction. After the extraction, the solution and residue are separated. The extraction residue then undergoes a second extraction, and the resulting extraction solution is combined with the first extraction solution where it is then concentrated and made into an extraction paste. It is then vacuum dried, pulverized, sieved, and analyzed before being added to the final blend. The Dry Pine Bark undergoes extraction, evaporation, separation of impurities, and filtration before being added to an absorption column for proanthocyanidins and polyphenols. After the column it is subjected to an ethanol wash, evaporation, spray drying, sieving, and analysis before being added to the final blend. Once each of the ingredients is added to the final blend, the nutraceutical is packaged and analyzed.DETAILED DESCRIPTION

[0015] The present invention may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the Examples included therein and to the Figures and their previous and following description.I. Definitions

[0016] To facilitate an understanding of the principles and features of the various embodiments of the disclosure, various illustrative embodiments are explained herein. Although exemplary embodiments of the disclosure are explained in detail, it is to be understood that other embodiments are contemplated. Accordingly, it is not intended that the disclosure is limited in its scope to the details of construction and arrangement of components set forth in the description or examples. The disclosure is capable of other embodiments and of being practiced or carried out in various ways.

[0017] In describing the exemplary embodiments, specific terminology will be resorted to for the sake of clarity. As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural references unless the context clearly dictates otherwise. For example, reference to a component is intended also to include composition of a plurality' of components. References to a composition containing “a” constituent is intended to include other constituents in addition to the one named.

[0018] Ranges may be expressed herein as from “about” or “approximately” or “substantially” one particular value and / or to “about” or “approximately” or “substantially” another particular value. When such a range is expressed, other exemplary embodiments include from the one particular value and / or to the other particular value.

[0019] The term “effective amount” as used herein is intended to mean the amount of the composition administered to achieve and maintain a sustained muscle pump in an individual. The effective amounts of the agent in the composition of the invention are effective over a wide dosage range and are generally administered in a nutraceutically effective amount. It will be understood, however, that the effective amount of the compound actually administered will be determined in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the age, weight, and response of the individual, and the like.

[0020] The term “bioactive ingredient” as used herein is intended to mean naturally occurring compounds which in an effective amount provide sustained muscle pump in an individual.. It will be understood that other bioactive ingredientscan be used that are capable of inducing a desired response or treating a particular condition.

[0021] By “comprising’’ or “containing” or “including” is meant that at least the named compound, element, particle, or method step is present in the composition or article or method, but does not exclude the presence of other compounds, materials, particles, method steps, even if the other such compounds, material, particles, method steps have the same function as what is named.

[0022] As used herein a “concentrate” refers to dried powder, slurry or suspension derived from a component that does not include the use of any solvents during the concentration process.

[0023] The term “dosage unit” is understood to mean a unitary', i.e. a single dose which is capable of being administered to a subject or individual, and that may be readily handled and packed, remaining as a physically and chemically stable unit dose comprising either the active ingredient as such or a mixture of it with solid or liquid nutraceutical vehicle materials. Dosages can be, but are not limited to oral, nasal, enteral, parenteral, transdermal, transmucosal, rectal, opthalmic, vaginal, etc.

[0024] The term “extract” refers to those substances prepared using a solvent, e.g., ethanol, water, steam, superheated water, methanol, hexane, chloroform liquid, liquid CO2, liquid N2, propane, supercritical CO2, or any combination thereof.Extracts, as used herein, can refer to an extract in a liquid form, or can refer to a product obtained from further processing of the liquid form, such as a dried powder or other solid form. Extracts may take many forms including but not limited to: solid, liquid, particulate, chopped, distillate, etc. and may be performed by any number of procedures or protocols, such as chopping, grinding, pulverizing, boiling, steaming, soaking, steeping, infusing, applying a gas, etc., and may employ any suitable reagents, such as water, alcohol, steam, or other organic materials. Extracts ty pically have a given purity percentage and can be relatively to highly pure. In some embodiments, extracts can be phytoextracts made from specific parts of a source, such as the skin, pulp, leaves, flowers, fruits of a plant etc., or can be made from the whole source. In some aspects an extract may include one or more active fractions or active agents. In some extracts, maltodextrin can be added as a carrier. In some aspects, thepurity of an extract can be controlled by, or be a function of the extraction process or protocol.

[0025] As used herein, '‘formulation” and ‘'composition” can be used interchangeably and refer to a combination of at least two ingredients. In some embodiments, at least one ingredient may be an active agent or otherwise have properties that exert physiologic activity when administered to a subject.

[0026] As used herein, “nutraceutically acceptable” refers generally to materials which are suitable for administration to a subject in connection with an active agent or ingredient. For example, a “nutraceutically acceptable carrier” can be any substance or material that can be suitably combined with an active agent to provide a composition or formulation suitable for administration to a subject. Excipients, diluents, and other ingredients used in or used to prepare a formulation or composition for administration to a subject can be used with such term.

[0027] As used herein the term “primary therapeutic agent” designates the presence of a therapeutic agent in a composition at an amount greater than the total combined amount of the extracts providing a combined or synergistic effect in the composition.

[0028] The term, “subject,” “subjects,” “subjects in need thereof,” and “individuals” includes humans as well as non-human subjects. It will be understood that the subject to which a compound of the invention is administered need not suffer from a specific traumatic state. Indeed, the compounds of the invention may be administered prophy lactically. The terms “subject,” “subjects,” “subjects in need thereof,” and “individuals” can be used interchangeably herein.

[0029] As used herein, “substantial” or “substantially” when used in reference to a quantity or amount of a material, or a specific characteristic thereof, refers to an amount that is sufficient to provide an effect that the material or characteristic was intended to provide. The exact degree of deviation allowable may in some cases depend on the specific context. Similarly, “substantially free of’ or the like refers to the tack of an identified element or agent in a composition. Particularly, elements that are identified as being “substantially free of’ are either completely absent from the composition, or are included only in amounts which are small enough so as to have no measurable effect on the composition.

[0030] As used herein, the term “solvent” refers to a liquid of gaseous, aqueous or organic nature possessing the necessary characteristics to extract solid material from a plant product. Examples of solvents would include, but not limited to, water, steam, superheated water, methanol, ethanol, ethyl acetate, hexane, chloroform, liquid CO2, liquid N2, propane, or any combinations of such materials.

[0031] The phrase “effective amount,” “therapeutically effective amount,” or “therapeutically effective rate(s)” of an active ingredient refers to a non-toxic, but sufficient amount or deliver}' rates of the active ingredient, to achieve therapeutic results in treating a disease or condition for which the ingredient is being delivered. It is understood that various biological factors may affect the ability of a substance to perform its intended task. Therefore, an “effective amount,” “therapeutically effective amount, “or “therapeutically effective rate(s)” may be dependent in some instances on such biological factors. Further, while the achievement of therapeutic effects may be measured by a physician or other qualified medical personnel using evaluations known in the art, it is recognized that individual variation and response to treatments may make the achievement of therapeutic effects a subjective decision. The determination of a therapeutically effective amount or delivery rate is well within the ordinary skill in the art of nutraceutical sciences and medicine.

[0032] The term “therapeutic,” “therapeutically,” and the like are used to encompass therapeutic, palliative, and / or prophylactic uses.

[0033] The terms “treat,” “treating,” or “treatment” as used herein and as well understood in the art, mean an approach for obtaining beneficial or desired results, including without limitation clinical results in a subject being treated. Beneficial or desired results can include, but are not limited to, alleviation or amelioration of one or more signs or symptoms of a condition, diminishment of extent of disease, stabilizing (i.e. not worsening) the state of a disease or condition, delaying or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission (whether partial or total), whether detectable or undetectable. “Treat,” “treating.” and “treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment and can be prophylactic. Such prophylactic treatment can also be referred to as prevention or prophylaxis of a disease or condition. The prophylaxis may be partial or complete. Partial prophylaxis may result in the delayed onset of a physiological condition. Theperson skilled in the art will recognize that treatment may, but need not always, include remission or cure.

[0034] As use herein, “administering” a compound or agent to a subject refers to imparting the compound or agent to the subject in a way that provides the subject with a positive physiologic or health benefit as a result of the administration. A number of specific administration routes are known, such as oral, enteral, transdermal, transmucosal, parenteral, intravenous, and injectable administration.

[0035] As used herein, “co-administer,” “co-administration,” and “coadministering,” refer to administering two or more active agents, compounds, extracts, supplements, etc. in a way that allows the active agents, compounds, extracts, supplements, etc. to have a concomitant or overlapping physiological effect on a subject. As such, co-administration includes both administration of multiple agents to a subject at the same time or within a time relative to one another that allows the physiologic or in-vivo effect or result of each agent to take place in a simultaneous or overlapping manner.

[0036] As used herein, “compounds” may be identified either by their chemical structure, chemical name, or common name. When the chemical structure, chemical name, or common name conflict, the chemical structure is determinative of the identity of the compound. The compounds described herein may contain one or more chiral centers and / or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i.e., geometric isomers), enantiomers or diastereomers. Accordingly, the chemical structures depicted herein encompass all possible enantiomers and stereoisomers of the illustrated or identified compounds including the stereoisomerically pure form (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures. Enantiomeric and stereoisomeric mixtures can be resolved into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the skilled artisan. The compounds may also exist in several tautomeric forms including the enol form, the keto form, and mixtures thereof. Accordingly, the chemical structures encompass all possible tautomeric forms of the illustrated or identified compounds. The compounds described also encompass isotopically labeled compounds where one or more atoms have an atomic mass different from the atomic mass conventionally found in nature. Examples of isotopes that may be incorporatedinto the compounds of the invention include, but are not limited to,2H,3H,13C,14C,15N,18O.17O, etc. Compounds may exist in un-solvated forms as well as solvated forms, including hydrated forms and as N-oxides. In general, compounds may be hydrated, solvated or N-oxides. Certain compounds may exist in multiple cry stalline or amorphous forms. Also contemplated are congeners, analogs, hydrolysis products, metabolites, and precursor or prodrugs of the compound. In general, all physical forms are equivalent for the uses contemplated herein and are intended to be within the scope of the present disclosure.

[0037] Comparative terms such as “more effectively,” “greater than,” “improved,” “enhanced,” “increased,” “decreased,” and like terms can be used to state a result achieved or property present in a formulation or process that has a measurably better or more positive outcome than the thing to which comparison is made. For example, when referring to an improved health of a subject the comparison may be made to the health of the subject prior to engaging in the activity7which improved the health of the subject. Moreover, such terms can be used to qualify a property of a component in a composition, a biological response, or a result that is measurably different from the same component when administered individually, a biological response that is markedly different from the expected biological response without the administration of a component or a composition, or a result that is measurably different from the results that occur using known methods. In one example, “increased or decreased concentration, secretion, or biosynthesis,” means an appreciable or measurable increase or decrease in amount (e.g. by at least 3%), concentration, rate of secretion or amount of biosynthesis of the referent compound.

[0038] IL Compositions

[0039] Athletes who perform rigorous endurance exercises must train to cope with the varying levels of fatigue for improving their physical activity7abilities. Overloading the body with increased training can cause muscle soreness and stiffness, increased oxidative stress, muscle cramps, and even immunosuppression (Suzuki K, et al. Med Set Sports Exerc. 2003, 35: 348-355; Suzuki K, et al. J Appl Physiol 1 85. 1999, 87: 1360-1367 ). While these problems can be prevented by establishing an appropriate training and reconditioning program, it has been reported that incorporating dietary supplements containing antioxidants improves performance andrecovery, endurance and recovery of post-exercise muscle performance (Crum EM, et al. J Int Soc Sports Nutr, 2017. 14: 14; Torregrosa-Garcla A, et al. Nutrients, 2019, 1 1: 721; Castell LM, Newsholme EA. Nutrition, 1997, 13: 738-742).

[0040] This disclosure provides nutraceutical compositions for promoting improvements in muscle performance including muscle strength and endurance.A. Compositions for enhancing muscle pump

[0041] Provided herein are nutraceutical compositions comprising mango fruit powder, black ginger extract, pine bark extract, and gingko biloba extract powder. The nutraceutical compositions support anti-inflammatory activities, notably through the modulation of nitric oxide synthase (NOS) pathways, thereby promoting an optimal environment for sustained muscle pump and performance enhancement.1. Mango fruit powder

[0042] Mango (Mangifera indicci L.) is one of the most important fruits worldwide due to its nutritional value and content of diverse phytochemicals, with diverse functions (Brecht and Yahia, CAB International, Wallingford, UK, Second Edition, pp. 484-528 (2009); Gupta et al., Bioscience. 48: 101783 (2022); Postharvest Biology and Technology of Tropical and Subtropical Fruits. Volume 1. Fundamental Issues, Woodhead Publishing, England (2011), pp. 21-78).

[0043] Some of the main bioactive compounds identified in mango fruit include phenolic acids (coumaric acid, ferulic acid and hydroxybenzoic acid), polyphenols (quercetin, mangiferin, catechins, tannins, kaempferol, anthocyanins, gallic acid, ellagic acid), carotenoids, which are the most abundant, and the vitamins ascorbic acid, thiamine, riboflavin, and niacin (Burton-Freeman et al., Food Funct., 8 (2017), pp. 3010-3032; Wall-Medrano et al., Nutr. Hosp., 31 (2015), pp. 67-75 ). These compounds have been reported to exhibit antioxidant activity (Zapata-Londono et al., Vitae, 27 (1) (2020)), and contribute to prevention of inflammatory processes (Xu et al., Eur. J. Cancer Prev., 22 (6) (2013)). These effects have been attributed to activation by antioxidants of signaling pathways able to regulate the expression of genes and activate molecules leading to antioxidant, immunoregulatory, antiproliferative, and antidiabetic effects (Yahia et al. Arabian Journal of Chemistry, 16(7): 104860 (2023).

[0044] The mechanisms of action explaining the bioactivity of several compounds present in mango fruit are based on in vitro, in vivo and clinical trials, butoften at very high concentrations that cannot be achieved by dietary means because these bioactive compounds are found in low levels in plant-based diets, thus unrealistically high intake would be necessary to achieve the levels found to be active using in vivo experiments. However, an active component can be overlooked in an extract or the interaction between several compounds that may be necessary' for a strong effect. In addition, it was often observ ed that the mixture (e.g.. an extract) shows a stronger response than adding the single active components together, reason for which the regular consumption of fruits, vegetables and whole grains is associated with reduced risk of developing chronic diseases in spite of the low abundance of some of their bioactive components (Pezzuto and Vang, Chemo prevent! on, Springer, Switzerland (2020), pp. 613-629 , Yahia et al., Postharvest physiology and biochemistry of fruits and vegetables, Woodhead Publishing, United Kingdom (2019), pp. 19-45 ).2. Black ginger extract

[0045] Black ginger is called Kaempferia parviflora and has the alias of black turmeric or Krachaidam. The plant belongs to the genus Zinguberaceae Kaempferia distributed in Southeast Asia. It is known as a health food in traditional medicine such as Thailand and Laos, and is said to have effects such as energy' enhancement and nutrition and tonic. Black ginger has been used as a dietary supplement in various preparations such as medicinal liquor or liquor plus honey, pills (powdered rhizome with honey), capsules, and tablets.

[0046] A large number of recent studies have demonstrated the biological activities of black ginger extract (Kaempferia parviflora extract: KPE) and polymethoxyflavones (PMFs) including anti-oxidative activity, etc (Toda K, et al. Heliyon. 2016 May 24;2(5):e001 15.). Clinical studies have shown KPE to improve physical fitness performance (Promthep K., et al. Med. Sci. Monit. Basic Res. 2015;21: 100-108; Wattanathom J., et al. Evid. Based Complement. Alternat. Med. 2012:2012:732816). Toda K et al. previously reported that PMFs in KPE increased energy production through AMP-activated protein kinase activation induced improvements of metabolism in myocytes (Toda K, et al. Heliyon. 2016 May 24;2(5):e00115). A powdered black ginger extract can be found in the market as a healthy food ingredient that has been standardized to contain not less than 2.5% of 5,7-dimethoxyflavone and 10% of total PMFs.

[0047] Toda et al. reported the beneficial effects of black ginger on fatigue or muscular endurance (Toda K, et al. Heliyon. 2016 May 24;2(5):e00115). The study- showed that KPE increases physical fitness performance and muscular endurance. Th results of the study7agreed with linical studies on black ginger reporting improved physical fitness performance in athletes, (Promthep K., et al. Med. Sci. Monit. Basic Res. 2015;21: 100-108; Wattanathom J., et al. Evid. Based Complement. Alternat. Med. 2012;2012:732816) and the enhanced antioxidant enzyme activities (increased activities of superoxide dismutase, glutathione peroxidase and catalase in serum) and decreased Malondialdehyde (MDA) level had been suggested to be one of the mechanisms for physical improvement effect of KPE.3. Pine bark extract

[0048] Pine bark extract has been shown to offer several natural and healthy ways to enhance sport endurance (Blazso G, et al. Phytother Res. 2004;18:579- 581; Peng QL, et al. Mol Brain Res. 2002;104:55-65; Rohdewald P. et al. IntJ Clin Pharmacol Ther. 2002;40: 158-168) and improve post-exercise recovery (Packer L, et al. Free Rad Biol Med. 1999;27:704-724) not only in professional athletes but also in people who participate in sports recreationally. Pine bark extract in sports nutrition is used to improve endurance, recovery7, cramps, and enhance blood flow.

[0049] Pine bark extract has been reported to have multiple physiological benefits for the athlete, including cardio-vascular and cholesterol lowering benefits, the ability to enhance microcirculation by increasing capillary- permeability, significant free radical scavenging activity- against reactive oxygen and nitrogen species, the potential to regenerate the ascorbyl radical and to protect endogenous vitamin E and glutathione from oxidative stress, and the potential to protect erythrocytes in G6PD deficiency (Gulati OP. et al. Biom Rev. 2005;16:49-57)..

[0050] Pinus pinaster is a French coastal pine bark extract rich in antioxidants (Igawa T, et al. J Phys Ther Sci. 2021 Apr;33(4):339-344). Recent studies demonstrate that Pinus pinaster was used by amateur athletes for reducing muscle pain and stiffness and improving running performance (Vinciguerra G, et al. Angiology1, 2006, 57: 331-339; Vinciguerra G, et al. Panminerva Med, 2019, 61 : 457-463; Vinciguerra G, et al. J Sports Med Phys Fitness, 2013, 53: 644-654). Igawa T et al. reported that endurance professional athletes using Pinus pinastersupplements demonstrated improvements in muscle pain, local muscle performance including muscle strength and endurance, and the amount of training.

[0051] Pinus massoniana Lamb is another specie of pine native to south and southwest of China. It’s bark, pollen, turpentine, and needles have been used in traditional Chinese medicine for the treatment of rheumatic arthralgia, hypertension, neurasthenia and chilblain (H.Y. Zhang, Key Records of Chinese Traditional Medicine in China, Science Press. Peking (1994); Y. Cui et al.. Phytother Res, 19 (1) (2005), pp. 34-38). In preclinical studies, Pinus massoniana bark extract (PMBE) has been reported reduce oxidative stress (M. Wang, et al. Am J Chin Med, 38 (5) (2010), pp. 909-919).

[0052] Pinus massoniana L. contains complex plant compounds known as proanthocyanidins. Proanthocyanidins are condensed tannins that are responsible for astringent character of fruits, berries, beans, and tea. These bioflavonoids are powerful antioxidants, giving benefits to the cardiovascular system and capillary health. They are potent, free radical scavengers that help the body reduce oxidative damage. Pinus massoniana has also been shown to be beneficial to sports nutrition for providing a source of energy, recovery, cramps, and better blood flow.4. Ginkgo biloba extract powder

[0053] Folium Ginkgo consists of the dried whole leaf of Ginkgo biloba.Ginkgo biloba is thought to have neuroprotective properties; however, the exact neuroprotective mechanism of Ginkgo biloba is not known, but Ginkgo biloba herbal extract components include flavone glycosides (which is made up of quercetin, kaempferol, rutin and myricetin) as well as terpene lactones (ginkgolides A and B), all of which decrease free radical release. In addition, terpene lactones have been shown to improve blood flow and reduce thrombus formation by inhibiting plateletactivating factor. Without being bound by any one theory, one mechanism of action by which compounds of the disclosure functions is by removing free radicals which cause oxidative damage to tissue. For example, Ginkgo biloba may reduce or prevent cell membrane lipid peroxidation, decrease oxidative damage to red blood cells and protect nerve cells. Thus, Ginkgo biloba herbal extract may benefit any cell or tissue that can be damaged by free radicals.

[0054] B. Mechanism of action1. Nitric oxide synthesis (NOS) pathway

[0055] The nutraceutical compositions provided herein include components that support anti-inflammatory activities, notably through the modulation of nitric oxide synthase (NOS) pathways, thereby promoting an optimal environment for sustained muscle pump and performance enhancement.

[0056] Certain nutritional compositions include nitric oxide or substances that increase or sustain the concentration of nitric oxide in the body. Nitric oxide is believed to have various functionalities in tissues and systems within the body including vascular tone, blood pressure, muscle contraction, muscle growth and response to contractile activity. (Sessa, W. C.. et al.. ‘"Regulation of endothelial derived nitric oxide in health and disease,’’ Mem. Inst. Oswaldo Cruz, 2005, 100: p. 15-18; Smith, L. W., et al., “Involvement of nitric oxide synthase in skeletal muscle adaptation to chronic overload,” J. Appl. Physiol., 2002, 92: p. 2005-2011.) Increasing nitric oxide production within the endothelium of vascular tissue may cause vasodilation and thereby may enhance muscle growth.

[0057] In the area of sports nutrition, nutritional compositions may include a source of the amino acid L-arginine or similar substances. It is believed that L- arginine is directly converted to nitric oxide in the body via the nitric oxide synthase (“NOS”) family of enzymes. (Boger, R. H., et al., “Clinical Pharmacology of L- Arginine,” Ann. Rev. Pharmacol. Toxicol., 2001 , 41 : p. 79-99.) Many nitric oxide sports nutrition products thus include an arginine ingredient, typically arginine alphaketogluterate. The nitric oxide produced from arginine may stimulate vasodilation.

[0058] Nitric oxide is an endogenously produced gaseous signaling molecule in the body. It is unique in its ability to be produced by one cell, then penetrate cellular membranes and regulate the functioning of other cells. Nitric oxide is typically formed from the amino acid precursor L-arginine by the enz me system that is known as nitric oxide synthase (“NOS”).

[0059] Nitric oxide possesses many functions within the body's tissues and systems including a role in vascular tone, blood pressure management, muscle contraction, muscle grow th and cell kinetics (response to contractile activity). Endothelial cells also contribute in the maintenance of vascular tone and structure. Nitric oxide, one of the major vaso-active mediators in endothelial tissue, typicallyincreases nitric oxide production within the endothelium of vascular tissue causing vasodilation (or vaso-relaxation) thereby increasing the "Pump" effect. Within the sports nutrition community, in particular bodybuilding, greater blood flow equals greater “Pump.” Through a synergistic combination of ingredients, the nutraceutical compositions described herein promote prolonged vasodilation through reducing the enzymes available responsible for reducing the muscle pump, allowing for extended muscle pump duration beyond what traditional supplements offer.2. Anti-inflammatory activity

[0060] Provided herein are nutraceutical compositions that inhibit inducibile NOS (iNOS), thereby reducing inflammation and the overproduction of NO in unwanted areas while upregulating endothelial NOS (eNOS) to increase NO production where it is beneficial, supporting sustained muscle pump.

[0061] The inflammatory response to exercise-induced muscle damage has been extensively described (Liao P., et al. Am. J. Physiol. Regul. Integr. Comp.Physiol. 2009;298:R599-R607). Exercise has been shown to have important modulatory effects on immunocyte dynamics, and possibly on immune function mediated by diverse factors, including exercise-induced release of pro-inflammatory cytokines, classical stress hormones, and hemodynamic effects leading to cell redistribution (Pedersen B.K., et al. Physiol. Rev. 2000;80: 1055-1081). Scientific literature has reports on the implication of several cytokines in the pathogenesis of muscle wasting and sarcopenia, most notably TNF-a, but also IL- 1 , IL-6, interferon (IFN)-y, and transforming grow th factor (TFG)- (Cai D., et al. Cell. 2004 ;119:285- 298).

[0062] Muscle fiber damage due to strenuous exercise, especially lengthening contraction, can trigger the release of inflammatory cytokines from immune cells and / or damaged muscle tissues. Gomez-Cabrera et al. reported that blood-bome polymorphonuclear (PMN) plays a critical role in defending the cell from viral and bacterial invasion by activating reduced nicotinamide adenine phosphate (NAD(P)H) oxidase to produce reactive oxygen species (ROS) via a respiratory burst (Gomez- Cabrera M.C., et al. Phys. Sportsmed. 2009;37: 116-123). During the early phase of muscle injury, inflammatory cytokines promote the gene expression of adhesion molecules such as VCAM-1. cytokine-induced neutrophil chemoattractant- 1 (CINC- 1), monocyte chemoattractant protein-1 (MCP-1). and NO. Liao P.. et al. reported thatsome cytokines can bind with membrane receptors and activate specific ROS- generating enzymes, such as NAD(P)H oxidase, and xanthine oxidase (XO) (Liao P., et al. Am. J. Physiol. Regul. Integr. Comp. Physiol. 2009;298:R599-R607).Endothelial cells from injured muscle are known to secrete TNF-a, IL-1, IL-6, and IL- 8, providing a positive feed-forward cycle (Pedersen B.K., et al. Can. J. Physiol.Pharmacol. 1998;76:505-511). Thus, oxidative stress and inflammation are closely linked.

[0063] Interactions between oxidative stress and inflammation been shown, indicating that the use of antioxidants can modulate inflammation in skeletal muscle. In vivo studies have shown that a high vitamin E diet prevents the exercise-induce increase in CINC-1 and MCP-1 levels and nuclear p65 content in rat gastrocnemius muscle (Powers S.K., et al. Compr. Physiol. 2011 ;1 :941-969). More recently the use of vitamin C (Thompson D., et al. Eur. J. Appl. Physiol. 2003;89:393-400), flavonoids and anthocyanins (Howatson G., et al. Scand. J. Med. Sci.Sports. 2009;20:843-852; Seeram N.P., etal. Phytomedicine. 2001;8:362-369) have been tested as potential treatments to reduce the oxidative stress and inflammatory responses after exhaustive exercise to accelerate recovery (Bell P.G., et A. Nutrients. 2014;6:829-843).III. Methods for prolonging muscle pump

[0064] Provided herein are methods of prolonging the duration of a muscle pump through natural vasodilation and blood flow enhancement. Muscle pump is a desirable condition for athletes and fitness enthusiasts, associated with an increased blood flow to the muscles, resulting in improved nutrient delivery, waste removal, and a visually enhanced appearance. Traditional supplements focus on the temporary induction of this state. However, there remains a significant need for a formulation capable of not only inducing but also extending the duration of a muscle pump to maximize these benefits over time.A. Formulations

[0065] This nutraceutical composition described herein may be administered in a wide variety of product forms including non-enteric nutraceutical dosage forms such as compressed and molded tablets, hard gelatin capsules, soft elastic gelatin capsules, and microcapsules that dissolve in the stomach, emulsions, and suspensions, or as partof a beverage or solid food product. The latter may be used as a meal supplement or replacement.

[0066] The nutraceutical composition formulations provided herein comprise 32-57% mango fruit powder, 23-36% black ginger extract, 13-25% pine bark extract, and 10-21% gingko biloba extract powder.

[0067] The composition according to the present invention may be formulated for administration by any suitable route such as the oral, rectal, nasal, topical (dermal) or parenteral administration route. Thus, the composition may be in the form of tablets, capsules, suspensions, emulsions, solutions, injectables, suppositories, sprays, aerosols and in other suitable form.

[0068] Formulations for oral use include tablets which contain the composition in a mixture with non-toxic nutraceutically acceptable excipients. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium chloride, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, potato starch or alginic acid; binding agents, for example, starch, gelatin or acacia; and lubricating agents, for example, magnesium stearate, stearic acid or talc. Other nutraceutically acceptable excipients can be colorants, flavoring agents, plasticizers, humectants etc. The tablets may be uncoated or they may be coated by known techniques, optionally to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed.

[0069] Formulations for oral use may also be presented as chewing tablets or chewing gum, or as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil.

[0070] Powders, dispersible powders or granules suitable for preparation of an aqueous suspension by addition of water are also convenient dosage forms of the present invention. Formulation as a suspension provides the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents are, for example, naturally- occurring phosphatides, as e.g. lecithin, or condensation products of ethylene oxidewith e.g. a fatty acid, a long chain aliphatic alcohol or a partial ester derived from fatty acids and a hexitol or a hexitol anhydrides, for example, polyoxyethylene stearate, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitan monooleate etc. Suitable suspending agents are, for example, sodium carboxymethylcellulose, methylcellulose, sodium alginate etc.

[0071] The formulation and preparation of the above-mentioned compositions is well-known to those skilled in the art of nutraceutical formulation. Specific formulation can be found in “Remington's Pharmaceutical Sciences.” (Remington's Pharmaceutical Sciences, 1980, 16thEd. Mack Publishing Company, Eason, USA)

[0072] Preferably, the nutraceutical composition of the present invention comprises a combination containing the bioactive ingredients: mango fruit powder, black ginger extract, pine bark extract, and gingko biloba extract powder. The ingredients may be distributed among a variety of dosage forms administered so as to deliver an effective amount of the composition of the invention. In the methods of the invention, the ingredients of the composition may be taken in a single dosage form. In other embodiments of the methods, the ingredients comprising an effective amount may be distributed in multiple dosage forms provided that the dosage forms are simultaneously or near-simultaneously administered.V. Administration

[0073] The nutraceutical compositions provided herein may be administered to a subject to enhance physical performance, specifically by prolonging the duration of a muscle pump through natural vasodilation and blood flow enhancement.

[0074] The nutraceutical formulation provided herein may also be administered parenterally (intravenous, intramuscular, subcutaneous or the like) in dosage forms or formulations containing conventional, non-toxic nutraceutically acceptable carriers and adjuvants. The formulation and preparation of such compositions is well-known to those skilled in the art of pharmaceutical and nutraceutical formulation. (Remington's Pharmaceutical Sciences, 1980, 16thEd. Mack Publishing Company, Eason, USA.)

[0075] For parenteral use, the nutraceutical compositions according to the invention may comprise the compounds in the form of a sterile inject on. To prepare such a composition, the compounds are dissolved or suspended in a parenterally acceptable liquid vehicle. Among acceptable vehicles and solvents that may beemployed are water, water adjusted to a suitable pH by addition of an appropriate amount of hydrochloric acid, sodium hydroxide or a suitable buffer, 1,3-butanediol, Ringer's solution and isotonic sodium chloride solution. The aqueous formulation may also contain one or more preservatives, for example, methyl, ethyl or n-propyl p- hydroxy benzoate.

[0076] For the rectal application, suitable dosage forms for a composition according to the present invention include suppositories (emulsion or suspension type), and rectal gelatin capsules (solutions or suspensions). In a typical suppository formulation, the thermogenic compounds are combined with an appropriate nutraceutically acceptable suppository base such as cocoa butter, esterified fatty' acids, glycerinated gelatin, and various water-soluble or dispersible bases like polyethylene glycols and polyoxyethylene sorbitan fatty acid esters. Various additives like e.g. enhancers or surfactants may be incorporated.

[0077] For the nasal application typical dosage forms for a composition according to the present invention include nasal sprays and aerosols for inhalation. In a typically nasal formulation, the active ingredients are dissolved or dispersed in a suitable vehicle. The nutraceutically acceptable vehicles and excipients and optionally other nutraceutically acceptable materials present in the composition such as diluents, enhances, flavoring agents, preservatives etc. are all selected in accordance with conventional nutraceutical practice in a manner understood by the persons skilled in the art of formulating nutraceuticals.

[0078] The nutraceutical compositions according to the invention may also be administered topically on the skin for percutaneous absorption in dosage forms or formulations containing conventionally non-toxic nutraceutically acceptable carriers and excipients including microspheres and liposomes. The formulations include creams, ointments, lotions, liniments, gels, hydrogels, solutions, suspensions, pastes, plasters and other kinds of transdermal drug delivery' systems. The nutraceutically acceptable carriers or excipients may include emulsifying agents, antioxidants, buffering agents, preservatives, humectants, penetration enhancers, chelating agents, gel forming agents, ointment bases, perfumes and skin protective agents.

[0079] Examples of emulsify ing agents are naturally occurring gums, e.g. gum acacia or gum tragacanth, naturally occurring phosphatides, e.g. soybean lecithin and sorbitan monooleate derivatives. Examples of antioxidants are butylated hydroxyanisole (BHA), ascorbic acid and derivatives thereof, tocopherol and derivatives thereof and cysteine. Examples of preservatives are parabens and benzalkonium chloride. Examples of humectants are glycerin, propylene glycol, sorbitol and urea. Examples of penetration enhancers are propylene glycol, DMSO, triethanolamine, N,N-dimethylacetamide, N,N-dimethylformamide, 2-pyrrolidone and derivatives thereof, tetrahydrofurfuryl alcohol and Azone™. Examples of chelating agents are sodium EDTA. citric acid and phosporic acid.

[0080] Examples of gel forming agents are Carbopol, cellulose derivatives, bentonit, alginates, gelatin and PVP.

[0081] Examples of ointment bases are beeswax, paraffin, cety l palmitate, vegetable oil. sorbitan esters of fatty acids (Span), polyethyleneglycols, and condensation products between sorbitan esters of fatty acids and ethylene oxide, e.g. polyoxyethylene sorbitan monooleate (Tween).EXAMPLESExample 1: Compositions

[0082] Materials and Methods

[0083] Provided herein are compositions and methods for making a proprietary^ blend of nutraceutical compositions for the product ENDOFLO™. Table 1 shows the specifications of the compositions. The compositions are not manufactured using any ingredient / component of animal origin. Nor do the compositions contain alcohol (<5000 ppm). Therefore, the compositions are suitable for vegan and vegetarian diets.

[0084] The compositions are not genetically modified and there is no contact with genetically modified organisms (GMOs) through the process, or through processing aids or additives.

[0085] Table 1: Composition specifications.

[0086] The compositions are provided in formulations as shown in Table 2.

[0087] Table 2: ENDOFLO

[0088] The process for making the nutraceutical compositions, also know n as ENDOFLO™ compositions, is shown in Figure 1. These compositions aid in extending "muscle pump” following exercise.

[0089] Example 2: Effect of ENDOFLO™ on exercise- induced hemodynamics.

[0090] This study aimed to elucidate the impact of ENDOFLO™ on skeletal muscle vasodilation and the temporal dynamics of muscle pump response following resistance exercise. The primary endpoints included alterations in post-exercise brachial circumference, hemodynamic fluctuations, and subject perception of muscle engorgement.

[0091] Materials and Methods:

[0092] Subjects adhered to a 72-hour washout period, abstaining from all dietary supplements except the investigational product. Standardized hydration and dietary intake were maintained, with a mandatory exclusion of caffeiene and stimulants on the day of testing. Participants refrained from structured exercise for 24 hours prior to assessment to minimize confounding variables related to muscular fatigue. A crossover methodology was employed, wherein each subject completed both a control session, and an experimental session, following a pharmaceutically effective amount of ENDOFLO™ administration. ENDOFLO™ was ingested 45 minutes pre-exercise, and basal brachial circumference and resting heart rate were recorded. Participants executed a structured bicep curl protocol with precisely controlled repetition schemes and inter-set rest intervals which included a warm-up phase of three sets, and a primary exercise phase including four sets. In the warm-up phase, the first set consisted of 12 repetitions at 30% 1 rep max weight (1RM), with 30-60 seconds of inter-set rest. The second set was 8 repetitions at 50% 1RM with 30-60 seconds of inter-set rest. The third set was of 4 repetitions at 70% 1RM with 60 seconds of interset rest. The primary exercise phase included 4 sets of 8 repetitions at 75-80% 1RM, with 2-minute inter-set intervals. Measurements of brachial circumference and heart rate were again obtained at predefined points. Subjective assessment of muscular engorgement, recorded as subjective pump ratings, and vascular reactivity' were collected at 5-minute intervals over a 60-minute post-exercise windoyv. Values for the measured variables are included in Table 3.

[0093] Table 3: Measured effect of ENDOFLO™ on brachial circumference, heart rate and subjective pump rating.

[0094] Independent-sample t-tests were conducted to evaluate inter-group differences and are included in Table 4.

[0095] Table 4: Statistical analysis of inter-group differences (* denotes statistical significance at a< 0.05)

[0096] Results:

[0097] The experimental group treated with ENDOFLO™ yielded a marginally greater increase in post-exercise brachial circumference compared to the control; however, this difference did not reach statistical significance (p=0.472), suggesting that acute hemodynamics shifts were comparable between conditions. Cardiovascular response was measured and evaluated through heart rate fluctuations. There was no significant difference with the administration of ENDOFLO™ (p = 0.797), indicating a neutral impact on systemic autonomic regulation. The subjective perception of muscle pump intensity was significantly enhanced in the group that received ENDOFLO™ (p = 0.0009), supporting its potential efficacy in augmenting muscle fullness and vascular reactivity post-exercise. Overall, while ENDOFLO™ did not elicit statistically significant changes in brachial circumference or cardiovascular parameters, it significantly enhanced perceived muscle pump duration and fullness.

[0098] The complete disclosure of all patents, patent applications, and publications, and electronically available material (including, for instance, nucleotide sequence submissions in, e.g., GenBank and RefSeq, and amino acid sequencesubmissions in. e.g., SwissProt, PIR. PRF, PDB, and translations from annotated coding regions in GenBank and RefSeq) cited herein are incorporated by reference. In the event that any inconsistency exists between the disclosure of the present application and the disclosure(s) of any document incorporated herein by reference, the disclosure of the present application shall govern. The foregoing detailed description and examples have been given for clarity of understanding only. No unnecessary limitations are to be understood therefrom. The invention is not limited to the exact details shown and described, for variations obvious to one skilled in the art will be included within the invention defined by the claims.

Claims

What is claimed is:

1. A nutraceutical composition for modulating the nitric oxide synthase (NOS) pathways comprising: a) mango fruit powder; b) black ginger extract; c) pine bark extract; and d) ginkgo biloba extract powder; wherein modulation of the NOS pathway supports anti-inflammatory activities and vasodilation to enhance muscle activity.

2. The nutraceutical composition of claim 1, wherein the mango fruit powder is a powder of aMangifera indica L fruit.

3. The nutraceutical composition of claim 1, wherein the black ginger extract is extracted from a Kaempferia parviflora root.

4. The nutraceutical composition of claim 1, wherein the pine bark extract is extracted from a I’ inns massoniana Lamb or a Pinus pinaster bark.

5. The nutraceutical composition of claim 1, wherein the ginkgo biloba extract powder is a powder of a dried whole leaf of Ginkgo biloba.

6. The nutraceutical composition of claim 1, wherein the composition is comprised of 32-57% mango fruit powder, 23-36% black ginger extract, 13-25% pine bark extract, and 10-21% gingko biloba extract powder.

7. A method of modulating the nitric oxide synthase (NOS) pathways of a subject comprising administering to the subject the nutraceutical composition of claim 1. wherein modulating the NOS pathways enhances a subject’s physical performance by prolonging the duration of a muscle pump.

8. The method of claim 7. wherein modulating the nitric oxide synthase (NOS) pathways enhances the subject’s vasodilation, blood flow, and antioxidant levels.

9. The method of claim 7. wherein modulating the nitric oxide synthase (NOS) pathways supports improved nutrient delivery and metabolic waste removal over an extended period.

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