Composition for stimulating nitric oxide production

Abstract

The present invention relates to a new composition for activating extraoral bitter taste receptors to produce nitric oxide, which comprises beetroot extract, hops, cinchona bark extract (which contains food-grade quinidine), L-arginine, phenolic extract of black pepper and oligomeric procyanidins of unripe apple, which combines multiple pathways for generating and preserving nitric oxide in the organism, improving the bioavailability thereof. The combination of the ingredients has a synergistic effect never before described in the prior art. The beetroot extract is the base for the nitrate substrate, and the oligomeric procyanidins of unripe apple are anti-inflammatory and promote endothelial health. The cinchona bark extract (which contains food-grade quinidine) and the hops are stimulants for nitric oxide production. The phenolic extract of black pepper helps the ingredients penetrate through the intestine or skin, and the L-arginine is a substrate for nitric oxide production. The composition can be formulated for oral or topical administration. The invention also describes a method for increasing nitric oxide production in an individual by using the composition.

Description

[0001] COMPOSITION ADMINISTERED TO STIMULATE NITRIC OXIDE PRODUCTION

[0002] TECHNICAL FIELD OF THE INVENTION

[0003] The present invention belongs to the technical field of medicinal preparations containing active organic ingredients, as it is a novel composition administered orally or topically to activate extraoral bitter receptors for nitric oxide production, comprising beetroot extract, hops, cinchona bark extract (containing food-grade quinidine), L-arginine, phenolic extract of black pepper, oligomeric procyanidins from unripe apple, which combines multiple pathways for the generation and preservation of nitric oxide in the body, improving its bioavailability.

[0004] BACKGROUND

[0005] Nitric oxide (NO) is a gaseous signaling molecule critical in multiple human physiological systems. It is synthesized endogenously from L-arginine by the action of nitric oxide synthases (NOS) [i]. It provides multiple benefits to the individual, for example, it supports blood vessel health, promotes the healing process and improves microcirculation to treat problems such as varicose veins and erectile dysfunction, increases the immune system's response to infections, regulates appetite, aids vasodilation, and provides anti-inflammatory benefits.

[0006] In the cardiovascular system, NO acts as a key vasodilator, relaxing vascular smooth muscle and modulating arterial tone [2]. It also inhibits platelet aggregation and reduces leukocyte adhesion, thus preventing thrombotic and inflammatory events [3].

[0007] In the immune system, NO generated by the inducible NOS isoform (NOS) in macrophages and dendritic cells plays direct antimicrobial and antiviral roles [4]. Furthermore, it modulates the inflammatory response, facilitating the resolution of the infectious process [5].

[0008] In the respiratory system, NO produced by the airway epithelium contributes to antimicrobial defense, regulation of ciliary motility, and adjustment of pulmonary vascular tone [6]. Nasal NO production, in particular, is a key mechanism for maintaining upper airway sterility.

[0009] In wound healing processes, NO is essential in the inflammation, proliferation, and remodeling phases, stimulating angiogenesis and collagen synthesis, for example. Reduced NO levels are associated with poor healing, as occurs in chronic wounds or in diabetic patients.

[0010] It is because of these multiple health benefits that nitric oxide (NO) has been extensively studied as a potential pharmaceutical agent. Typically, to induce nitric oxide production in individuals, they are provided with an amino acid substrate that includes arginine and ornithine. However, there is also a genomic pathway for producing NO in response to extraoral bitter receptors, which has not historically been utilized.

[0011] In the prior art, only one international patent application, published as WO2020245573A1, is known, which describes a combination, kit, or composition comprising: (I) one or more nitrite salts; (II) a proton source comprising one or more acids selected from organic carboxylic acids and organic non-carboxylic reducing acids; and (III) one or more organic polyols. When one or more nitrite salts react with the proton source in the presence of the one or more organic polyols, the combination, kit, or composition yields reaction products that include nitric oxide, optionally other nitrogen oxides, and / or optionally precursors thereof, and which are useful, for example, in the treatment of various disorders.

[0012] Therefore, it would be ideal to find a composition, preferably of natural ingredients, that can be administered orally or topically, to stimulate NO production in the individual.

[0013] OBJECT OF THE INVENTION The first object of the present invention is a novel composition for activating extraoral bitter receptors for nitric oxide production, comprising beetroot extract, hops, cinchona bark extract (containing food-grade quinidine), L-arginine, phenolic extract of black pepper, and oligomeric procyanidins from unripe apple. This composition combines multiple pathways for NO generation and preservation in the body, enhancing its bioavailability. The composition can be formulated for oral or topical administration. A second object of the invention is a method for stimulating nitric oxide production in an individual using this composition.

[0014] DESCRIPTION OF THE INVENTION

[0015] The characteristic details, examples, and embodiments of the invention are clearly shown in the following description and accompanying figures, which are numbered to indicate the parts shown. However, it is clear and apparent to a person skilled in the art that the invention is not limited to these embodiments and can be adapted to various applications.

[0016] The composition of the present invention can be used to stimulate NO production in an individual and may comprise the following ingredients, which are provided as a reference and do not limit the composition to additional ingredients. A person skilled in the art should understand that there are equivalent elements that can be substituted without changing the essential function or operation of the composition.

[0017] The composition comprises beetroot extract, hops, cinchona bark extract (containing food-grade quinidine), L-arginine, phenolic extracts of black pepper, and oligomeric procyanidins from unripe apple. These are all-natural ingredients with no side effects, unlike the international patent application published as WO2020245573A1, whose composition consists primarily of salts, acids, and polyols. The composition can be administered orally as an encapsulated freeze-dried powder or topically as a cream or gel.

[0018] L-arginine is a direct precursor for NO synthesis via endothelial and neuronal NOS [8]. When administered orally, it improves endothelial function and circulation [8], when administered topically it promotes healing by stimulating NOS [9], and when administered intranasally it enhances local NO production in the respiratory mucosa

[10] .

[0019] Beetroot (Beta vulgaris) extract is rich in nitrates, which are converted to NO via the nitratonithto-NO pathway [n]. When administered orally, it improves muscle perfusion and reduces blood pressure

[0012] . When applied topically, it stimulates microcirculation and wound healing

[0013] . When administered intranasally, it promotes local NO, strengthening mucociliary defenses

[0014] .

[0020] Oligomeric procyanidins from unripe apples are potent antioxidants that protect endogenous NO and activate eNOS

[0015] . When administered orally, they increase the bioavailability of endothelial NO

[0015] , topically they protect against oxidative stress, promoting regeneration

[0016] , and intranasally they enhance TAS2R agonism, increasing local defenses

[0017] .

[0021] Hops (Humulus lupulus) contains bitter acids that activate TAS2R receptors and promote NO production [is]. When administered orally, it improves endothelial function and reduces oxidative stress [is], when applied topically it stimulates healing and has anti-inflammatory effects

[0019] , and when administered intranasally it activates the production of antimicrobial NO in the mucosa

[0020] .

[0022] Cinchona bark extract (containing food-grade quinidine) (bitter taste) is a potent TAS2R receptor agonist, inducing rapid NO release

[0021] . When administered orally, it activates vagal reflexes related to vascular relaxation

[0022] ; topically, it stimulates a local neurogenic response with potential NO release.

[0023] Intranasal administration induces NO release for respiratory defense

[0021] .

[0023] Black pepper extract (Piper nigrum or Piper longum) or pipehna improves the bioavailability of polyphenols and nitrates, also activates TRPV1

[0024] . When administered orally, it increases the absorption of L-arginine and polyphenolic compounds

[0024] , when administered topically it stimulates neurogenic vasodilation, improving microcirculation

[0025] Intranasal administration induces cleansing reflexes and potential NO release

[0026] .

[0024] The combination of ingredients has a synergistic effect never before described in the prior art. Beetroot extract is the base for the nitrate substrate, while oligomeric procyanidins from unripe apple are anti-inflammatory and promote endothelial health. Cinchona bark extract (containing food-grade quinidine) and hops stimulate nitric oxide production. The phenolic extract of black pepper aids in the penetration of the ingredients through the intestine or skin, and L-arginine is a substrate for nitric oxide production. The novel aspect of the composition is the effect of stimulating bitter taste receptors to accelerate the nitric oxide production immune response, through bitter ingredients such as hops and cinchona bark extract (containing food-grade quinidine).

[0025] In summary, the composition can have three main parts. First, there is the substrate for nitric oxide production, which may include beetroot extract, phenolic extract, and L-arginine. Next, there are the bittering agents, such as cinchona bark extract (containing food-grade quinidine) and hops, which can stimulate bitter taste receptors and accelerate the immune response, where nitric oxide plays an important role. Finally, there are the absorption components, such as black pepper, which can help promote and facilitate the absorption of the composition through the skin or intestines.

[0026] In different embodiments, the ingredients may be mixed as a dry powder, encapsulated for oral administration, or mixed into a topical base, such as a cream or gel for topical administration. The amount of the ingredients may vary depending on the route of administration. For example, some concentrations may be irritating to the skin but not to the digestive tract. The method for stimulating NO production in the individual comprises administering an effective therapeutic dose of a composition comprising beetroot extract, hops, cinchona bark extract (containing food-grade quinidine), L-arginine, phenolic extracts of black pepper, and oligomeric procyanidins from unripe apple, where the composition may be administered orally or topically.

[0027] Endogenous NO production can be enhanced through two main pathways: the classical pathway of NOS synthesis from L-arginine and the alternative nitrate-nitrite-NO pathway. Recently, activation of extraoral bitter taste receptors TAS2R has also been shown to induce local NO release.

[0028] PRACTICAL IMPLEMENTATION OF THE INVENTION

[0029] To demonstrate the efficacy of the formulation, a single-capsule formulation study was conducted. The specific objective of this study was to compare acute salivary NO responses between substrate-only formulations and TAS2-enhanced 2000 mg single-capsule formulations in healthy adults.

[0030] In summary, the methodology applied included twenty healthy participants who gave their verbal informed consent for this internal safety and efficacy evaluation. Subjects received either substrate-only capsules (L-arginine 750 mg + beetroot extract 1250 mg providing 25 mg of nitrate) or TAS2R-enriched capsules (L-arginine 245 mg + beetroot extract 1250 mg + oligomeric procyanidins 200 mg + hops 70 mg + cinchona bark extract (containing food-grade quinidine) 2.5 mg + pipehne 5 mg + cellulose 227.5 mg) in a randomized, double-blind design. Both formulations totaled exactly 2000 mg. Salivary NO was assessed using commercial colorimetric strips, with a standardized 6-point scale (from 0 = depleted to 5 = very high, and 3 = target representing optimal levels) at baseline and at 60 minutes. The following paragraphs provide a detailed description of the study.The enhancement of nitric oxide (NO) has traditionally relied on substrate supplementation via L-arginine and dietary nitrates. Recent evidence reveals that bitter taste receptors (TAS2R) in cardiovascular tissues trigger a rapid release of NO when activated by specific compounds. TAS2R agonists induce concentration-dependent vasorelaxation through NO signaling.

[0031] This study compares conventional substrate supplementation against an enhanced formulation with TAS2R to assess whether multi-pathway activation can overcome reduced substrate availability, a result that would be unexpected based on current knowledge.

[0032] Study design and participants

[0033] Randomized, double-blind, controlled pilot study in 20 healthy adults (18-55 years, BMI 19-29 kg / m²) 2All participants gave their verbal informed consent for this internal evaluation of the safety and efficacy of the ingredients, having been informed about the nature of the ingredients and the procedures involved. Both formulations contained a total amount of exactly 2000 mg.

[0034] The group that only received substrate received capsules with 750 mg of L-arginine and 1250 mg of beetroot extract that provided 25 mg of nitrate.

[0035] The group with enhanced TAS2R received capsules containing 245 mg of L-arginine, 1250 mg of beetroot extract providing 25 mg of nitrate and 200 mg of phenolic extract obtained from unripe apple peels, 70 mg of hop extract, 2.5 mg of food-grade cinchona bark extract (containing food-grade quinidine), 5 mg of black pepper extract with at least 95% piperine, and 227.5 mg of microcrystalline cellulose.

[0036] Procedures: Initial saliva collection, consumption of a capsule with 250 ml of water after an overnight fast, second collection at 60 minutes. NO assessment was performed with commercial reagent strips using a standardized 6-point scale: 0 = Depleted, 1 = Low, 2 = Borderline, 3 = Target, 4 = High, 5 = Very High.

[0037] Statistical analysis

[0038] Mann-Whitney U test for changes in NO, chi-square for goal attainment, Cohen's d for effect size.

[0039] Results

[0040] Initial features

[0041] Groups matched by age (Substrate: 36.9 ± 4.8 years vs. TAS2R: 37.1 ± 4.8 years), sex (60% vs. 60% males) and baseline NO levels (1.3 ± 0.8 in both groups).

[0042] Main results

[0043] The substrate-only group showed an average improvement of +1.6 categories, and the target was achieved in 8 out of 10 (80%) of the participants, with a range of +1 to +2 categories.

[0044] The group with improved TAS2R showed an average improvement of +2.8 categories, and the target was achieved in 9 out of 10 (90%) of the participants, with a range of +1 to +3 categories.

[0045] Statistical analysis

[0046] The TAS2R formulation demonstrated superior efficacy (p<0.001). Surprisingly, despite a 67% reduction in L-arginine, the group with the enhanced treatment achieved a 75% improvement (more than 1.2 additional categories) with a very large effect size (Cohen's d = 2.76).

[0047] Table 1 below summarizes the results of the study. This result was unexpected and contradicts conventional understanding. Despite containing 67% less L-arginine (245 mg vs. 750 mg), the TAS2R-enhanced formula achieved a 75% greater increase in NO. This result would not have been predictable based on prior knowledge of L-arginine dose-response relationships, indicating an unprecedented synergistic effect.

[0048] Mechanistic Analysis Enhanced Bioavailability: Piperine (5 mg) increases the absorption of L-arginine by 100% to 200%, effectively converting 245 mg into 490-735 mg of bioavailable L-arginine, potentially equating the dose of substrate alone through increased absorption rather than a higher nominal content.

[0049] TAS2R-mediated NO release: Cinchona bark extract (containing food-grade quinidine) (2.5 mg, 5 mg maximum allowed according to supplement guidelines) and hop α-acids activate vascular TAS2R, triggering calcium-dependent eNOS activation, independent of substrate pathways.

[0050] This represents a previously unknown pathway for NO enhancement that works synodically with traditional substrate approaches, rather than simply additively to traditional substrate approaches.

[0051] Antioxidant protection of NO: The oligomeric procyanidins contained in the apple extract (200 mg) offer a double benefit: direct stimulation of endothelial NO synthase (eNOS) and protection of NO against oxidative degradation, thus prolonging the half-life of the NO generated.

[0052] Vascular optimization: Hop compounds enhance endothelium-dependent relaxation through multiple ion channels, creating optimal conditions for NO activity and bioavailability.

[0053] Unexpected results and not obvious

[0054] The superior performance of the formulation with reduced L-arginine demonstrates an effect that would not be obvious to a subject matter expert. Conventional belief predicts that reducing the primary substrate of nitric oxide (NO) by two-thirds would decrease, rather than improve, the biological response. The 75% improvement despite the 67% reduction in substrate indicates that the combination produces more than the sum of its parts—a classic definition of unexpected synergy.

[0055] Clinical Implications: The 90% response rate represents a significant improvement over conventional supplementation. Administering a single capsule enhances therapeutic adherence while maintaining superior efficacy within established safety parameters for supplements.

[0056] Limitations of the study

[0057] Given the small sample size (n = 10 per group), these findings should be considered exploratory; however, the magnitude of the observed effect (Cohen's d = 2.76) and the consistency of the response (90% vs 80% of responders) suggest a strong trend that warrants follow-up studies with larger samples.

[0058] Conclusion

[0059] A single 2000 mg capsule incorporating TAS2R receptor agonists achieved a 75% greater improvement in nitric oxide (NO) than the substrate-only formulation, through an unexpected synergistic mechanism that compensates for the reduced L-arginine content. This result would not have been predictable based on existing knowledge, as it contradicts the assumption that NO improvement is primarily substrate-limited. Demonstrating superior performance with a substantially lower primary substrate indicates a non-obvious advance that warrants patent protection.

[0060] These findings validate multi-pathway NO enhancement strategies and support progression to larger controlled trials with cardiovascular endpoints.

[0061] The embodiments of the invention described above are for illustrative and non-limiting purposes; a person skilled in the art should recognize that the invention can be carried out in other specific forms without departing from its essence, which is not limited by the illustrative details of the description but by the scope of the claims.

[0062] References

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Claims

CLAIMS Having sufficiently described my invention, I consider it novel and therefore claim as my exclusive property the content of the following claims:

1. A composition that can be administered to an individual to stimulate the production of nitric oxide, characterized in that it comprises: a) A substrate; b) Bitterness stimulants for bitter taste receptors; and c) An absorption enhancer.

2. The composition according to claim 1, wherein the substrate comprises beetroot extract, a phenolic extract and L-arginine.

3. The composition according to claim 2, wherein the beetroot extract has a nitrate content of at least 2%.

4. The composition according to claim 2, wherein the phenolic extract is obtained from unripe apple peels.

5. The composition according to claim 2, wherein the phenolic extract is obtained from grape seeds or pine bark.

6. The composition according to claim 1, wherein the bitterness stimulants for bitter taste receptors comprise cinchona bark extract (containing food-grade quinidine) and hops, wherein the cinchona bark extract (containing food-grade quinidine) comprises food-grade cinchona bark extract (containing food-grade quinidine).

7. The composition according to claim 1, wherein the absorption enhancer comprises a phenolic extract of Piper nigrum.

8. The composition according to claim 7, wherein the Piper nigrum has a purity of 95%.

9. The composition according to claim 1, wherein the absorption enhancer comprises a phenolic extract of Piper longum.

10. The composition according to claim 7, wherein the Piper longum has a purity of 95%.

11. A composition, administerable to an individual, for stimulating nitric oxide production, characterized in that it comprises: a) Beetroot extract; b) Phenolic extract of unripe apple peels; c) Cinchona bark extract (containing food-grade quinidine); d) Hops extract; e) Phenolic extract of Piper nigrum or Piper longum; and f) L-arginine.

12. The composition of claim 11, administered orally in capsules, characterized in that it comprises: a) 1250 to 5000 mg of beetroot extract; b) 75 to 500 mg of phenolic extract of unripe apple peels; c) 0.25 to 2.5 mg of food-grade cinchona bark extract (containing quinidine); d) 25 to 75 mg of hop extract; e) 1.25 to 5 mg of phenolic extract of Piper nigrum or Piper longum; and f) 125 to 1500 mg of L-arginine.

13. The composition of claim 11, for topical administration, characterized in that it comprises: a) 1.0 to 3.0% by weight of beetroot extract; b) 0.2 to 1.5% by weight of phenolic extract of unripe apple peel; c) 0.01 to 0.1% by weight of food-grade cinchona bark extract (containing quinidine); d) 0.2 to 0.8% by weight of hop extract; e) 0.005 to 0.05% by weight of phenolic extract of Piper nigrum or Piper longum; and f) 5.0 to 20.0% by weight of L-arginine.

14. The composition of claim 11, administered by aerosol, characterized in that it comprises: a) 2.0 to 5.0% by weight of beetroot extract; b) 0.3 to 2.0% by weight of phenolic extract of unripe apple peel; c) 0.005 to 0.05% by weight of food-grade cinchona bark extract (containing quinidine); d) 0.3 to 1.2% by weight of hop extract; e) 0.002 to 0.02% by weight of phenolic extract of Piper nigrum or Piper longum; and f) 3.0 to 12.0% by weight of L-arginine.

15. A method for stimulating nitric oxide production in an individual, characterized in that it comprises supplying the composition of any of claims 11 to 14.

16. The use of the composition of any of claims 1 to 14 to be administered to an individual to stimulate nitric oxide production.

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