Formulation and composition of a natural senolytic agent

Abstract

The present invention relates to a formulation and composition of a natural senolytic agent for the selective elimination of senescent cells and anti-aging applications. The composition comprises three synergistic groups of plant-derived active ingredients: (1) core senolytics in the proanthocyanidin group, (2) cellular sensitizers, and (3) anti-inflammatory and senescence- associated secretory phenotype (SASP) inhibitors. These components act in concert to induce senescent cell clearance with efficacy comparable to anticancer agents such as Navitoclax and the combination of Dasatinib and Quercetin. The formulation also demonstrates mitochondrial protective effects, reduces oxidative stress and melanin synthesis, and enhances skin hydration and rejuvenation. Composed entirely of safe, naturally derived compounds, the invention is suitable for integration into cosmetic products, dietary supplements, and other health care applications, and holds strong potential for development into pharmaceutical products targeting age-related pathologies.

Description

[0001] FORMULATION AND COMPOSITION OF A NATURAL SENOLYTIC AGENT Field of the Invention

[0002] The present invention relates to the fields of biotechnology and gerontology, and more particularly to the formulation and composition of a natural senolytic agent.

[0003] Background of the Invention

[0004] Cellular senescence is recognized as one of the twelve fundamental hallmarks of aging. It refers to a state in which cells in the body cease to divide and are termed 'senescent cells' (commonly referred to as "zombie cells"). These senescent cells can secrete factors associated with a senescence-associated secretory phenotype (SASP), which promote chronic inflammation, damage surrounding tissues, and accelerate the aging process. Consequently, the accumulation of senescent cells contributes to the development of various degenerative diseases, such as type 1 and type 2 diabetes, Parkinson's disease, dementia, heart failure, liver fibrosis, osteoarthritis, atherosclerosis, osteoporosis, cancer, and skin aging.

[0005] Senolytics are a class of substances, including both synthetic drugs and naturally derived compounds, that selectively eliminate senescent cells by inducing apoptosis or through other mechanisms such as disruption of survival pathways or oxidative stress overload. Substances effective in eliminating senescent cells are often repurposed anticancer agents or experimental compounds. For example, Navitoclax (ABT-263) induces apoptosis by inhibiting anti-apoptotic proteins BCL-2, BCL-XL, and BCL-w; Dasatinib, a tyrosine kinase inhibitor, disrupts multiple intracellular signaling pathways such as the Src family and PI3K / Akt; and UBX0101 interferes with protein-protein interactions, including the p53 / MDM2 axis.

[0006] Based on current scientific knowledge, researchers have developed treatments for age-related diseases using senolytic agents known as “senolytic therapeutics.” In studies with aged rhesus macaques, the administration of Navitoclax significantly reduced biomarkers of cellular senescence and inflammation — particularly the senescence-associated secretory phenotype (SASP) — in the central nervous system. Improvements in synaptic function and reductions in neuronal damage markers were also observed. Although thrombocytopenia (reduced platelet count) occurred as a side effect, no serious adverse events were reported. These findings highlight the potential of Navitoclax as a senolytic therapeutic for alleviating neurodegenerative decline.

[0007] A pilot clinical study evaluated the combination of Dasatinib (100 mg / day) and Quercetin (1250 mg / day), referred to as D+Q, in 14 patients diagnosed with Idiopathic Pulmonary Fibrosis (IPF). The agents were administered three consecutive days per week over a three-week period. The treatment led to improvements in physical performance, including six-minute walk distance, walking speed, and chair-stand time. Reported adverse effects were generally mild to moderateand included nausea, headache, coughing, and dyspnea. These findings suggest the potential of senolytic therapy to alleviate functional symptoms in patients with IPF.

[0008] Furthermore, a Phase 2 randomized, double-blind, placebo-controlled clinical study investigated the combination of Dasatinib and Quercetin (D+Q) in patients with Alzheimer’s disease. Preliminary findings in individuals with mild cognitive impairment (MCI) or early-stage Alzheimer’s revealed that D+Q penetrated the central nervous system, reduced the burden of senescent cells, and lowered levels of SASP-related inflammatory markers. The treatment also showed trends toward reduced accumulation of tau protein and amyloid-beta — hallmark proteins of Alzheimer’s pathology. Reported adverse effects were generally mild, including fatigue, nausea, and headache. These findings suggest that D+Q may hold promise as a novel senolytic approach for managing Alzheimer’s disease, although further validation in larger studies is needed.

[0009] However, the aforementioned pharmaceutical agents cannot be incorporated into cosmetic or dietary supplement products due to regulatory limitations and the potential for adverse side effects with long-term use. Consequently, research efforts have increasingly focused on identifying naturally derived senolytic compounds, which exhibit promising potential for use in consumer products intended for regular application, such as cosmetics and dietary supplements. These natural compounds are generally considered safe based on historical usage and are aligned with prevailing consumer preferences for naturally sourced ingredients.

[0010] Examples of plant-derived extracts that have been studied for their potential to eliminate senescent cells include Quercetin (found in apples, shallots, and green tea) and Fisetin (present in strawberries, apples, and cucumbers). However, the senolytic efficacy of these individual compounds remains limited compared to certain anticancer drugs. For example, Quercetin typically requires co-administration with Dasatinib to achieve meaningful senolytic activity.

[0011] Recognizing these limitations, the inventors developed a natural senolytic formulation comprising active and synergistic compounds derived from plant sources. This formulation effectively eliminates senescent cells, demonstrating efficacy comparable to pharmaceutical agents such as Navitoclax and the Dasatinib-Quercetin combination. It is suitable for use in cosmetic, dietary supplement, and other health care applications, and also holds potential for pharmaceutical development. Additionally, the formulation offers superior anti-aging and skinrejuvenating effects compared to commercially available cosmetic active ingredients.

[0012] A review of both Thai and international patent databases revealed prior disclosures related to inventions concerning the formulation and composition of a senolytic agent. The details of such inventions are summarized as follows.U.S. Patent Publication No. US 8974839B2, titled “Dietary Supplement System for Multifunctional Anti-Aging Management and Method of Use,” discloses an oral dietary supplement composed of three synergistic complexes: (a) a telomere maintenance complex comprising Purslane extract, turmeric root, quercetin dehydrate, cayenne pepper fruit, vanadyl sulfate (a form of vanadium), fenugreek seed, and omega fatty acids; (b) a calorie restriction and gene expression enhancement complex containing trans-resveratrol (from Polygonum cuspidatum root), pterostilbene, fisetin, alpha-lipoic acid, and coenzyme Q10; and (c) a free radical scavenging complex including green tea leaf extract, catechins and polyphenols, bilberry-derived anthocyanins, and grape skin extract. The formulation is intended to promote systemic anti -aging effects and support cellular function.

[0013] The Chinese patent publication No. CN117721161A, titled “Bio-based Multifunctional Anti-Aging Additive as well as Preparation Method and Application Thereof,” discloses that the bio-based anti-aging additive comprises polymers synthesized from phenolic compounds. The preparation involves dissolving a phenolic compound in a solvent, followed by the addition of radish peroxidase and hydrogen peroxide (H2O2) to initiate a polymerization reaction. The mass ratio used in the reaction ranges from 100:3:39 to 100:8:200 (phenolic compound: peroxidase: H2O2). The phenolic compounds of biological origin include phenol, flavonoids, phenolic acids, coumarins, and anthocyanins. The flavonoids can be selected from one or more of the following: quercetin, luteolin, rutin, myricetin, epigallocatechin, kaempferol, puerarin, daidzin, fisetin, hesperetin, and genistein. The resulting additive exhibits excellent ultraviolet (UV) protection, oxidative and thermal stability, and is compatible with high molecular weight materials, thereby enhancing anti-aging performance effectively.

[0014] The U.S. patent publication No. US20170216286A1, titled “Killing Senescent Cells and Treating Senescence-Associated Conditions,” discloses the use of Quercetin in combination with a Src protein inhibitor for the elimination of senescent cells and the treatment of conditions associated with cellular senescence, such as osteoarthritis and chronic inflammatory diseases. The invention particularly emphasizes the therapeutic application of quercetin in conjunction with an active pharmaceutical ingredient, rather than its use as a standalone treatment.

[0015] The international patent publication No. WO2018215795A3, titled “Senolytic Compounds,” discloses the use of senolytic compounds for the elimination of senescent cells. The compounds described in the patent are predominantly synthetic or semi-synthetic pharmaceuticals, such as cardiac glycosides, focal adhesion kinase (FAK) inhibitors, 3 -hydroxy-3 -methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, as well as other agents including cyclosporine, rotenone, vincristine, and temsirolimus (an mTOR inhibitor). These compounds are primarilydesigned for pharmaceutical applications in the treatment of cardiovascular diseases, cancer, and aging-related conditions. The patent emphasizes the use of structurally specific drugs for senescent cell clearance, with no particular focus on naturally derived substances.

[0016] The European patent publication No. EP3643305A1, titled "Compositions for the Elimination of Senescent Cells," discloses compositions for the targeted removal of senescent cells through the use of inhibitors of cyclooxygenase- 1 (COX-1), cyclooxygenase-2 (COX-2), lipoxygenase, and acyl-CoA synthetase long-chain family member (ACSL). The compounds employed include aspirin, celecoxib, zileuton (a leukotriene synthesis inhibitor), and nordihydroguaiaretic acid (NDGA). These compounds act by modulating eicosanoid metabolism, including increasing intracellular arachidonic acid levels in senescent cells, thereby promoting their selective elimination. The patent primarily addresses therapeutic applications for diseases associated with cellular senescence, such as cardiovascular disorders, neurodegenerative diseases, and chronic inflammation. Notably, the claimed compositions focus on pharmaceutical agents, except for NDGA, which is a naturally derived compound.

[0017] The Thai petty patent application No. 1803002202, titled “Nanostructured Lipid Carrier Composition for Encapsulating Flavonoids Comprising Bile Salts to Enhance Intestinal Absorption and a Method for Preparing Such Particles” discloses a nanostructured lipid carrier (NLC) composition that demonstrates stability and enhanced intestinal absorption of active ingredients. The composition comprises 0.1-1% by weight of a flavonoid compound, 0.1-2.5% by weight of a solid lipid, 1-10% by weight of a liquid lipid, 6.5-30% by weight of a surfactant, and a pharmaceutically acceptable carrier adjusted to 100% by weight. A key innovation of this invention is the incorporation of bile salts into the surfactant system, which enhances the intestinal absorption of the flavonoid compound. The flavonoid used in the composition is Quercetin. The preparation method is designed to yield nanostructured lipid particles with high stability and significantly enhanced bioavailability of the encapsulated flavonoid, primarily through improved intestinal absorption.

[0018] It is evident that the prior art utilizes various classes of natural substances — such as flavonoids and polyphenols — as primary active agents or alternative compounds for anti-aging purposes, owing to their antioxidant and anti-inflammatory properties. These substances are commonly incorporated as key ingredients or additives in cosmetic or dietary supplement products to promote anti-aging effects and enhance antioxidant performance. Furthermore, there are instances where such compounds are combined with lipid-based nanoparticles to improve delivery efficiency in product formulations. In inventions specifically targeting senolytic properties for theelimination of senescent cells, the prior art predominantly involves the use of pharmaceutical drugs, synthetic compounds, or a combination of natural substances with pharmaceutical agents.

[0019] However, no prior art has been found that fully discloses the same technical features as the present invention in every aspect. The invention is distinct in terms of the formulation and composition of the senolytic agent, which is derived entirely from natural sources. This agent is suitable for use in cosmetic products, dietary supplements, other health care products, and even pharmaceuticals. It demonstrates senolytic efficacy comparable to that of Navitoclax and the combination of Dasatinib with Quercetin, in effectively eliminating senescent cells. Furthermore, the composition promotes cellular and skin rejuvenation. The invention comprises three synergistic groups of plant-derived substances: core senolytics in the proanthocyanidin group, cellular sensitizers, and anti-inflammatory and SASP inhibitors.

[0020] Summary of the Invention

[0021] The formulation and composition of a natural senolytic agent according to the present invention consist of three key components that synergistically enhance senolytic efficacy: (1) core senolytics in the proanthocyanidin group, (2) cellular sensitizers, and (3) anti-inflammatory and SASP (senescence-associated secretory phenotype) inhibitors. These components are derived from plant-based sources such as grape seeds, maritime pine bark, apples, strawberries, blueberries, onions, and other botanical materials. Each component is incorporated in optimized proportions to ensure maximum senolytic and anti -aging activity.

[0022] The objective of the present invention is to develop a formulation comprising active ingredients capable of effectively delaying aging and selectively eliminating senescent cells. Specifically, the invention employs the synergistic action of the three plant-derived compound groups, collectively referred to as “Active Z.” This formulation demonstrates selective senescent cell clearance efficacy comparable to potent anticancer agents such as Navitoclax and Dasatinib, and superior to previously reported natural compounds in the scientific literature. In addition, the formulation provides mitochondrial protection by mitigating oxidative stress, reduces reactive oxygen species (ROS) production, inhibits melanin synthesis, enhances skin hydration, and improves skin radiance. Crucially, the invention relies on safe and efficacious natural compounds or extracts, making it suitable for integration into a broad range of cosmetic products, dietary supplements, and health care applications.

[0023] Brief Description of the Drawings

[0024] Figure 1. Senolytic efficacy of Navitoclax, a commercially available cosmetic-grade senolytic active (Active AR), and Active Z. The test was conducted using primary human dermalfibroblasts (HDFs), comparing young (P3) and senescent (P9) cell populations to assess selective cell clearance.

[0025] Figure 2. Senolytic efficacy of Navitoclax, plant-derived compounds (Quercetin, Fisetin, and grape seed extract), two commercial dietary supplements (Supplements C and L), and Active Z, evaluated using human skin explants ex vivo skin model) under systemic treatment conditions. Test substances were introduced via the culture medium to simulate oral administration.

[0026] Figure 3. Comparative senolytic activity of Active Z, Retinol, and seven commercially available cosmetic active ingredients (M3000, MM, DM, PM, RN, SA, and RP), assessed in human skin explants ex vivo model) under topical application conditions to simulate cosmetic use.

[0027] Figure 4. Mitochondrial restoration activity of Active Z compared to Resveratrol in human dermal fibroblasts exposed to oxidative damage induced by hydrogen peroxide. Mitochondrial membrane potential (MMP) was assessed via TMRE fluorescence staining.

[0028] Figure 5. Skin rejuvenation efficacy of Active Z, Retinol, and seven commercial cosmetic actives (M3000, MM, DM, PM, RN, SA, and RP), evaluated by stimulation of collagen and hyaluronic acid production in human skin explants (ex vivo skin) under topical treatment conditions.

[0029] Detailed Description of the Invention

[0030] The formulation and composition of a natural senolytic agent for senescent cell elimination and anti-aging purposes comprises the following components:

[0031] A) core senolytics in the proanthocyanidin group in an amount of 42% to 92% by weight; B) cellular sensitizers in an amount of 4.0% to 33% by weight; and

[0032] C) anti-inflammatory and SASP inhibitors in an amount of 2.0% to 30% by weight.

[0033] The proanthocyanidins serve as the principal senolytic agent for inducing apoptosis in senescent cells. The proanthocyanidin can be selected from procyanidin Cl, procyanidin Bl, procyanidin B2, oligomeric proanthocyanidins (OPCs), grape seed extract, and maritime pine bark extract, either alone or in combination.

[0034] The proanthocyanidins are extracted from plant sources selected from the group consisting of grape seeds, maritime pine bark, cocoa seeds, apple peel, blueberry, cranberry, and strawberry, either alone or in combination.

[0035] The cellular sensitizers are adjuvant agents that enhance the susceptibility of senescent cells to apoptotic cell death, functioning by modulating the cellular survival pathways or by increasing the cells’ sensitivity to oxidative stress. The cellular sensitizers can be selected from the group consisting of piperlongumine, curcumin, gingerenone A, and quercetin, either alone or in combination.The cellular sensitizers are extracted from plant sources selected from the group consisting of long pepper (Piper longum), black pepper (Piper nigrum), turmeric Curcuma longa), zedoary (Curcuma zedoaria), shallot (Allium ascalonicum), onion (Allium cepa), ginger (Zingiber officinale), Sophora japonica, and apple (Malus domestica), either alone or in combination.

[0036] The anti-inflammatory and SASP inhibitors serve as adjuvant agents that mitigate inflammation associated with aging and reduce cellular and tissue damage caused by senescent cells. The anti-inflammatory and SASP inhibitors can be selected from the group consisting of apigenin, fisetin, kaempferol, ellagic acid, and ginsenosides, either alone or in combination.

[0037] The anti-inflammatory and SASP inhibitors are preferably extracted from plant sources selected from the group consisting of parsley, chamomile, strawberry, apple, smoke tree (Cotinus coggygria), broccoli, kale, pomegranate, raspberry, and ginseng, either alone or in combination.

[0038] Experimental Example 1: Efficacy Test of Navitoclax, Commercially Available Senolytic Actives, and Active Z in Eliminating Senescent Cells Using Primary Human Dermal Fibroblasts (HDFs) According to the Present Invention

[0039] As shown in Figure 1, the efficacy of the active compound in eliminating senescent cells was tested using human dermal fibroblast cells, specifically comparing young cells (HDF P3) with senescent cells (HDF P9), to assess its senolytic potential. The test groups included treatments with Dimethyl Sulfoxide (DMSO) as a negative control (serving as the solvent for the active compounds), Navitoclax at 1.2 pg / mL as a positive control, a commercially available cosmetic senolytic active referred to as Active AR at 6,250 pg / mL, and the disclosed invention’s active composition designated as Active Z at a concentration of 7 pg / mL. The cells were treated with each compound for a duration of three days, after which cell viability was measured using a standard cell viability assay. The results demonstrated that Active Z successfully eliminated up to 46% of the senescent cells, achieving efficacy comparable to Navitoclax and superior to Active AR, despite being applied at a concentration approximately 893 times lower than Active AR. These findings indicate that Active Z, derived from natural sources, possesses strong senolytic activity with high potency even at low concentrations.

[0040] Experimental Example 2: Efficacy Test of Navitoclax, Natural Active Compounds, Commercial Dietary Supplements, and Active Z in the Elimination of Senescent Cells Using an Ex Vivo Human Skin Model Under Systemic Treatment Conditions

[0041] As shown in Figure 2, the efficacy of Navitoclax, natural active compounds, commercially available dietary supplements, and Active Z in eliminating senescent cells was tested using ex vivo human skin tissue obtained from a 41 -year-old volunteer. The study was designed as a systemic treatment model by incorporating each test substance into the tissue culture medium, therebysimulating the systemic effect of oral administration of drugs or dietary supplements on skin tissue via the circulatory system. The objective was to assess the ability of each substance to induce apoptosis in senescent cells. The skin tissue was divided into several treatment groups: a negative control group treated with dimethyl sulfoxide (DMSO), used as the solvent for test compounds; a group treated with Navitoclax at a concentration of 2.43 pg / mL; a group treated with Dasatinib at 97.6 ng / mL in combination with Quercetin at 7.5 pg / mL; a group treated with Quercetin alone at 15 pg / mL; a group treated with Fisetin at 15 pg / mL; a group treated with grape seed extract at 15 pg / mL; groups treated with two commercial dietary supplements, referred to as Supplement C and Supplement L, each at 15 pg / mL; and a group treated with Active Z, a proprietary senolytic formulation disclosed in the present invention, also at 15 pg / mL. The treatment period lasted five days, after which the culture medium was analyzed for levels of the senolytic biomarker 15-deoxy-A12,14-prostaglandin J2. The results showed that Active Z significantly increased the secretion of this biomarker by up to 7429% compared to the DMSO control group, with no statistically significant difference from the Navitoclax group or the Dasatinib + Quercetin group. Moreover, Active Z induced a greater expression of the senolytic marker than other natural senolytic agents reported in the literature, including Quercetin, Fisetin, and grape seed extract. It also outperformed the tested commercial anti-aging supplements C and L, which are marketed for their claimed senolytic properties.

[0042] Experimental Example 3: Efficacy Test of Active Z in the Elimination of Senescent Cells Compared to Cosmetic Active Ingredients Using an Ex Vivo Human Skin Model Under Topical Treatment Conditions

[0043] As shown in Figure 3, the efficacy of Active Z in eliminating senescent cells was tested in comparison to various commercially available cosmetic active ingredients, using ex vivo human skin tissue obtained from a 45-year-old volunteer. The test was conducted via topical application, wherein each active substance was directly applied to the surface of the skin tissue to simulate conditions akin to actual use in cosmetic or topical pharmaceutical products. The skin tissue samples were divided into multiple groups: a negative control group treated with water and ethanol, a positive control group treated with retinol — an established anti-aging compound — and several groups treated with cosmetic active ingredients from international brands (identified as M3000, MM, DM, PM, RN, SA, and RP) at concentrations ranging from 2 to 10 mg / mL. Another group was treated with Active Z, the novel active substance disclosed in this invention, at a concentration of 2 mg / mL. The treatments were applied for 5 consecutive days, after which the culture medium was analyzed for the presence of the senolytic marker 15-deoxy-A12,14-Prostaglandin J2 (15-d-PGJ2). The results revealed that Active Z significantly enhanced thesecretion of 15-d-PGJ2 by up to 255-fold compared to the negative control group (water and ethanol), indicating a potent ability to induce senescent cell clearance. In contrast, none of the other commercial active ingredients tested induced an increase in 15-d-PGJ2 levels relative to the negative control. These findings demonstrate that Active Z exhibits a unique and superior ability to promote the elimination of senescent cells, a property not observed in conventional market-available anti-aging actives.

[0044] Experimental Example 4: Efficacy Test of Active Z in Mitochondrial Restoration Compared to Resveratrol Using Free Radical-Damaged Human Fibroblast Skin Cells

[0045] As shown in Figure 4, the efficacy of Active Z in mitochondrial recovery was tested and compared to that of resveratrol in primary human dermal fibroblasts subjected to oxidative stress. The fibroblast cells were treated with either 400 pM hydrogen peroxide (H2O2), representing oxidative damage, or with phosphate-buffered saline (PBS) as a control vehicle. The experimental groups included a negative control treated with dimethyl sulfoxide (DMSO), a positive control treated with 2.28 pg / mL resveratrol (a known mitochondrial protective anti-aging agent), and a test group treated with 7 pg / mL of Active Z, the active composition disclosed in this invention. All treatments were conducted over a period of 48 hours. Following the incubation, mitochondrial membrane potential (MMP) was assessed using tetramethylrhodamine ethyl ester (TMRE) dye staining. TMRE accumulates in functional mitochondria, producing red fluorescence; loss of membrane potential results in diminished red signal intensity. The results demonstrated that hydrogen peroxide caused oxidative damage leading to a significant reduction in mitochondrial membrane potential and TMRE fluorescence intensity. Treatment with Active Z was found to attenuate the damage and restore mitochondrial membrane potential up to 60%, a level comparable to that achieved with resveratrol. These findings indicate that Active Z exhibits substantial potential for protecting mitochondria against oxidative damage, with efficacy similar to that of resveratrol.

[0046] Experimental Example 5: Efficacy Test of Active Z in Skin Regeneration by Stimulating Collagen and Hyaluronic Acid Production Using an Ex Vivo Human Skin Model Under Topical Treatment Conditions

[0047] As shown in Figure 5, the skin rejuvenation efficacy of Active Z was tested through its ability to stimulate collagen and hyaluronic acid production using ex vivo human skin tissue obtained from a 45-year-old volunteer. The study employed a topical treatment model in which the active substances were applied directly to the surface of the skin explants, simulating cosmetic or dermatological product application for localized effects. The test groups included negative controls treated with water and ethanol, and a positive control treated with retinol — an established anti-aging agent — at a concentration of 2.5 mg / ml. Additionally, commercially available cosmetic actives from various international brands (designated as M3000, MM, DM, PM, RN, SA, and RP) were tested at concentrations of 2 mg / ml, 2.5 mg / ml, or 10 mg / ml. Active Z, the subject active agent of this invention, was tested at a concentration of 2 mg / ml. Following a 5-day treatment period, collagen content in the skin tissue was assessed via Masson-Trichrome staining, while hyaluronic acid levels in the tissue culture medium were quantified using an enzyme-linked immunosorbent assay (ELISA). The results demonstrated that Active Z stimulated collagen production in the dermal layer by 94%, comparable to leading market actives. Notably, hyaluronic acid synthesis increased by up to 484%, surpassing the efficacy of both retinol and all tested foreign anti-aging actives. These findings indicate that Active Z possesses superior anti-aging potential for rejuvenating aged skin.

[0048] Experimental Example 6: Skin Irritation Test of Active Z in Human Volunteers

[0049] Based on the results of a 24-hour occlusive human patch test conducted on 23 volunteers (7 males and 16 females), Active Z at a concentration of 0.2% exhibited a skin irritation score of 0. This indicates that Active Z at this concentration is safe for use and can be considered a nonirritating ingredient.

[0050] Example 7: Skin Irritation Test of Lipid-Based Nanoparticles Encapsulating Active Z in Human Volunteers

[0051] In a 24-hour occlusive human patch test conducted on 23 volunteers (comprising 4 males and 19 females), the lipid-based nanoparticles encapsulating Active Z at a concentration of 3% exhibited a skin irritation score of 0. This result indicates that the formulation is non-irritating to human skin. Therefore, the lipid-based nanoparticles encapsulating Active Z at a 3% concentration can be considered safe and non-irritating for topical application.

[0052] Experimental Example 8: Hypoallergenic Test of Active Z in Human Volunteers

[0053] Based on the Human Repeated Insult Patch Test (HRIPT) conducted on 61 human volunteers, including 17 males and 44 females, it was found that Active Z at a concentration of 0.2% did not induce any allergic reactions. Therefore, it can be considered hypoallergenic.

[0054] Experimental Example 9: Efficacy Test of Active Z for Skin Rejuvenation in Oral Supplement Form in Human Volunteers

[0055] A study was conducted in which 13 volunteers consumed 200 milligrams of Active Z daily for a period of 11 days. The results demonstrated a statistically significant improvement in skin hydration, as well as a significant increase in the Individual Typology Angle (ITA), indicating a reduction in skin pigmentation. No adverse events were reported during the study.It should be understood that modifications or variations of the invention can be readily apparent to those skilled in the art without departing from the scope and spirit of the invention, as defined by the appended claims.

[0056] Best Mode of the Invention

[0057] As described in the Detailed Description of the Invention.

Claims

Claims1. A formulation and composition of natural senolytic agent, comprising:• a core senolytic component in the proanthocyanidin group, in an amount of 42% to 92% by weight;• a cellular sensitizer component, in an amount of 4.0% to 33% by weight; and• an anti-inflammatory and senescence-associated secretory phenotype (SASP) inhibitor component, in an amount of 2.0% to 30% by weight.

2. The formulation according to claim 1, wherein the proanthocyanidins are selected from the group consisting of procyanidin Cl, procyanidin Bl, procyanidin B2, oligomeric proanthocyanidins (OPCs), grape seed extract, and maritime pine bark extract, either alone or in combination.

3. The formulation according to any one of claims 1 or 2, wherein the proanthocyanidins are extracted from plant sources selected from the group consisting of grape seeds, maritime pine bark, cocoa seeds, apple peel, blueberry, cranberry, and strawberry, either alone or in combination.

4. The formulation according to claim 1, wherein the cellular sensitizers are selected from the group consisting of piperlongumine, curcumin, gingerenone A, and quercetin, either alone or in combination.

5. The formulation according to any one of claims 1 or 4, wherein the cellular sensitizers are extracted from plant sources selected from the group consisting of long pepper, black pepper, turmeric, mango ginger, shallot, onion, ginger, Sophora japonica. and apple, either alone or in combination.

6. The formulation according to claim 1, wherein the anti-inflammatory and SASP inhibitors are selected from the group consisting of apigenin, fisetin, kaempferol, ellagic acid, and ginsenosides, either alone or in combination.

7. The formulation according to any one of claims 1 or 6, wherein the anti-inflammatory and SASP inhibitors are extracted from plant sources selected from the group consisting of parsley, chamomile, strawberry, apple, smoke tree (Cotinus coggygria), broccoli, kale, pomegranate, raspberry, and ginseng, either alone or in combination.

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