Composition derived from okra for enhancing stamina, process for its preparation and use thereof

Abstract

The present invention relates to a palatable, free-flowing powdered composition derived from Abelmoschus esculentus for enhancing endurance. The composition comprises polysaccharides (5-50%) including rhamnogalacturonans and arabinogalactans; flavonoid glycosides (2-10%) including quercetin 3-O-gentiobioside and isoquercitrin (Okraenduroside); and total carbohydrates (30-70%), along with optional excipients for stability and bioavailability. The composition, when administered to a subject in an appropriate dose, improves running endurance, enhances swimming endurance, improves rate of perceived exertion (RPE), ameliorates severity of fatigue, improves fatigue score, increases peak power, and enhances endurance and muscle performance. The invention also discloses a method of preparation involving aqueous-alcoholic extraction, concentration, and purification to obtain a water-soluble powder. The composition can be formulated into capsules, tablets, powders, and beverages and administered orally at a dose of 100-2000 mg / day. Animal and human studies confirm the efficacy of the composition for enhancing endurance, making it suitable for the nutraceutical, sports nutrition, and pharmaceutical industries.

Description

Technical Field

[0001] This invention relates to a composition derived from okra plants for enhancing endurance and reducing fatigue in test subjects. The invention also discloses a method for preparing the composition and its use for enhancing endurance. Background Technology

[0002] Okra, scientific name Abelmoschus esculentus Okra, belonging to the Malvaceae family, is commonly known as "beauty finger" okra and has several other common names in different geographical regions where it is cultivated, including yellow okra, six-cornered bean, yellow hibiscus, hairy eggplant, coffee okra, foreign pepper, and goat's horn pepper. The immature green pods of okra are widely consumed as a vegetable, and extracts from these pods are used as thickeners in many soup and sauce recipes to increase their viscosity. Generally, okra is a high-value crop because it represents a source of nutrients important for human health, such as vitamins, potassium, calcium, carbohydrates, dietary fiber, and unsaturated fatty acids such as linoleic acid and oleic acid. Durazzo et al. disclosed that extracts from young okra pods have also been reported to exhibit moisturizing and diuretic properties, while the seeds of this plant have been reported to have anticancer and antifungal properties. Durazzo et al. evaluated okra (… Abelmoschus esculentus The bioactive components and beneficial properties of okra, such as anti-diabetic activity, were disclosed. Dantas et al. disclosed the extraction of okra mucilage and its use as a thickener and food stabilizer. Xia Fangbo et al. disclosed antioxidant and anti-fatigue components from okra seeds and pericarps. Yan Li disclosed the composition and anti-fatigue activity of water-soluble okra derived from okra stem pectin. Existing technology

[0003] A review of existing technologies shows that, although okra ( Abelmoschus esculentus It has been studied for its anti-fatigue effects, but there is limited evidence for palatable powder compositions derived from okra for enhancing endurance, which are explicitly claimed in the patent.

[0004] It is worth noting that existing research has confirmed the antioxidant and anti-fatigue properties of okra seeds and pericarps, indicating their potential to reduce fatigue and improve endurance. However, they have not disclosed specific palatable powder formulations or detailed preparation methods aimed at enhancing endurance. Patent application number CN104223271A discloses an okra lactic acid bacteria fermented beverage, but it does not focus on enhancing endurance.

[0005] This invention discloses a novel palatable powder composition derived from okra, specifically designed to enhance endurance and reduce fatigue. This composition, its preparation method, and its uses differ from existing literature and patents. Summary of the Invention

[0006] This invention relates to a method derived from okra ( Abelmoschus esculentus This invention provides a novel, palatable, free-flowing powdered composition for enhancing endurance and reducing fatigue in subjects. The invention also provides methods for preparing and using this composition. The composition comprises polysaccharides, flavonoid glycosides, and carbohydrates, and is formulated to enhance endurance and reduce fatigue when administered in effective doses.

[0007] Key bioactive components in the composition include quercetin 3-O-gentiobiglycoside and isoquercetin (okraenduroside) as flavonoid glycosides, and rhamnose galacturonic acid polysaccharide and arabinogalactan as polysaccharides. These components play crucial roles in regulating adenosine receptors (A1 and A2A), AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor Delta (PPARδ), and nuclear factor erythroid 2-related factor 2 (Nrf2), thereby optimizing energy metabolism, reducing oxidative stress, and promoting sustained performance.

[0008] The invention is derived from okra ( Abelmoschus esculentus The composition for enhancing endurance and reducing fatigue comprises: a) a polysaccharide, said polysaccharide being in the range of 5% to 50%; b) flavonoid glycosides, said flavonoid glycosides being in the range of 2% to 10%; and c) carbohydrates, said carbohydrates being in the range of 30% to 70%. In an embodiment of the invention, the composition comprises: a) a polysaccharide, said polysaccharide being in the range of 5% to 50%, wherein at least a portion comprises rhamnose-galacturonic acid polysaccharide and arabinogalactan; b) flavonoid glycosides, said flavonoid glycosides being in the range of 2% to 10%, wherein at least a portion comprises quercetin 3-O-gentiobiglycoside and isoquercetin (okra flavonoid glycosides); and c) carbohydrates, said carbohydrates being in the range of 30% to 70%. In a preferred embodiment of the invention, the composition comprises: a) 36.7% total carbohydrates; b) 5% flavonoid glycosides; and c) 11.5% polysaccharide. Excipients may be added to the composition to enhance stability and bioavailability. The composition is a palatable, free-flowing, light brown, water-soluble powder that can be formulated into various dosage forms, including capsules, tablets, powders, teas, syrups, dispersions, solutions, sticks, chewable tablets, sports drinks, and dietary supplements. Furthermore, the composition can be used as a complex ingredient in other foods, pharmaceuticals, sports drinks, meal replacement drinks, ointments, or water additives. The composition is intended to be administered orally to subjects at an effective dose ranging from 100 mg / day to 2000 mg / day to achieve the desired endurance enhancement benefits.

[0009] As described above, administration of the okra extract composition to mammals results in: a) improved running endurance; b) enhanced swimming endurance; c) improved subjective physical performance (RPE); d) improved fatigue severity; e) improved fatigue score; f) increased peak power; and g) enhanced endurance and muscle performance.

[0010] The preparation method involves water-alcohol extraction of dried okra pods, followed by ethyl acetate purification, ethanol precipitation, and spray drying to obtain a water-soluble, palatable powder composition.

[0011] A method for preparing an okra-derived composition for enhancing endurance according to the invention, the method comprising the steps of: a) pulverizing dried okra pods; b) extracting the powder with a hydroethanol solution at 90°C for 90 minutes; c) repeating the process twice; d) combining all filtrates and concentrating at 70°C; e) adding ethyl acetate, stirring at room temperature for 2 minutes, and allowing to stand for 1 hour; f) separating the ethyl acetate layer and the aqueous layer; g) concentrating the aqueous layer at 85°C; h) adding ethanol and cooling the solution to 4°C for 24 hours; i) clarifying the solution, centrifuging, and collecting the centrifuged fraction; and j) concentrating the centrifuged fraction at 95°C and drying to obtain a free-flowing powder.

[0012] This invention provides a novel nutritional and sports nutrition solution that provides sustained energy, improves performance, and accelerates recovery, thereby benefiting athletes, fitness enthusiasts, and individuals who need to enhance endurance and stamina. Detailed Implementation

[0013] This invention discloses a method using okra ( Abelmoschus esculentus This invention discloses a palatable, free-flowing powder composition prepared from polysaccharides, flavonoid glycosides, and carbohydrates. The invention also discloses the ability of the composition, when administered in effective amounts, to enhance the endurance and reduce fatigue in subjects, and a method for preparing the palatable powder composition.

[0014] The powder composition of the present invention comprises quercetin 3-O-gentiobiglycoside and isoquercetin (collectively referred to as okra flavonoid glycosides) as flavonoid glycosides. The powder composition also comprises rhamnogalacturonic acid polysaccharide and arabinogalactan as polysaccharides.

[0015] okra ( Abelmoschus esculentus Okra is a flowering plant valued for its edible green pods. It is low in calories and a good source of dietary fiber. Okra also contains various vitamins and minerals, including vitamin C, vitamin K, and folic acid. One of the notable characteristics of okra is the mucilage (a sticky substance) released when the pods are cut or cooked. This mucilage has thickening properties and is often used in some traditional dishes. However, the viscous nature of okra mucilage can cause digestive discomfort in some individuals, leading to bloating or flatulence. In this invention, a palatable, free-flowing, light brown powder composition soluble in water is obtained from okra and used to enhance endurance.

[0016] The role of ATP in endurance enhancement: Endurance refers to the body's ability to sustain physical activity for extended periods or resist fatigue over a long time. It involves the cardiovascular, respiratory, musculoskeletal, and metabolic systems, which must work together to support prolonged or repetitive exercise. Endurance activities place high demands on the adenosine triphosphate (ATP) energy system, requiring the body to efficiently produce and regenerate ATP, which is crucial for sustained performance.

[0017] ATP is the primary energy currency within cells. The energy stored in ATP is used for various cellular processes, such as muscle contraction, active transport of ions across cell membranes, and the synthesis of macromolecules such as proteins and nucleic acids.

[0018] The energy in ATP is generated through cellular respiration, substrate-level phosphorylation, and the transfer of high-energy phosphate groups. ATPases (adenosine triphosphate enzymes) catalyze the hydrolysis of ATP into ADP (adenosine diphosphate) and inorganic phosphate (Pi), thereby releasing energy for cells to use for muscle contraction and other biological functions.

[0019] Adenosine A1 receptor proteins play a crucial role in regulating energy metabolism, fatigue, and endurance during physical activity. Activation of the A1 receptor leads to a decrease in cyclic adenosine monophosphate (cAMP) levels, which has an inhibitory effect on cellular activity, thereby promoting fatigue and reducing athletic performance. Quercetin 3-O-gentiobiose and isoquercetin, present in okra extract compositions, can potentially bind to the A1 receptor, blocking the fatigue-inducing effect of adenosine or modulating receptor activity, thereby reducing fatigue and enhancing endurance. The binding affinity of quercetin 3-O-gentiobiose is -10.2 kcal / mol, and that of isoquercetin is -8.7 kcal / mol. This low binding affinity indicates a strong interaction between quercetin 3-O-gentiobiose and the adenosine A1 receptor, which can lead to enhanced endurance and reduced fatigue.

[0020] Adenosine A2A receptors also play a crucial role in regulating energy metabolism, inflammation, and muscle function. Quercetin 3-O-gentiobiose and isoquercetin, derived from okra extracts of this invention, bind to and activate the adenosine A2A receptor, thereby enhancing endurance and reducing fatigue during physical activity. The binding affinity of quercetin 3-O-gentiobiose is -8.40 kcal / mol, and that of isoquercetin is -7.40 kcal / mol, indicating a strong binding interaction between these two compounds and the adenosine A2A receptor. A more negative binding affinity indicates a stronger and more stable interaction between the ligand (compound) and its receptor. By activating the adenosine A2A receptor, quercetin 3-O-gentiobiose and isoquercetin can promote better energy balance and metabolism during exercise, thereby helping the body conserve energy and delaying the onset of fatigue.

[0021] Quercetin 3-O-gentiobiose and isoquercetin, derived from okra extract, improve peroxisome proliferator-activated receptor δ (PPARδ), a key regulator of lipid metabolism, fatty acid oxidation, and muscle endurance. The binding affinity of quercetin 3-O-gentiobiose (-9.0 kcal / mol) and isoquercetin (-7.9 kcal / mol) indicates a relatively strong interaction between these two compounds and PPARδ. PPARδ activation promotes the use of fatty acids as the primary energy source, thereby conserving glycogen reserves and providing sustained energy supply during prolonged physical activity.

[0022] AMP-activated protein kinase (AMPK) is a key enzyme regulating energy balance, and its activation can enhance endurance, reduce fatigue, and improve overall physical performance. Quercetin 3-O-gentiobiose and isoquercetin, present in the okra extract composition, activate AMPK through multiple mechanisms, including increasing the AMP / ATP ratio and reducing oxidative stress. The binding affinity of quercetin 3-O-gentiobiose at -9.6 kcal / mol and that of isoquercetin at -9.2 kcal / mol indicates that these two compounds bind relatively strongly to AMPK. The negative binding energy values ​​within this range indicate a stable and favorable interaction between the compounds and AMPK, meaning that these compounds can effectively activate the receptor. AMPK activation increases energy production, enhances lipid oxidation, and promotes mitochondrial biosynthesis, all of which support improved endurance during prolonged physical activity. By optimizing energy use, promoting effective recovery, and reducing oxidative stress, quercetin and isoquercetin help delay fatigue and improve overall athletic performance.

[0023] Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) plays a key role in enhancing endurance and reducing fatigue by improving the body's ability to efficiently produce and utilize energy. Quercetin 3-O-gentiobiose and isoquercetin, present in the okra extract composition, significantly enhance endurance and reduce fatigue by activating PGC-1α. The binding affinity of both quercetin 3-O-gentiobiose and isoquercetin is -8 kcal / mol, indicating that both compounds exhibit moderate to strong binding affinity to PGC-1α. PGC-1α activation promotes more efficient energy production and better mitochondrial function, enabling muscles to sustain longer periods before reaching fatigue. PGC-1α activation also helps reduce inflammation, which helps alleviate the soreness and discomfort often associated with strenuous physical activity, further reducing fatigue during exercise.

[0024] Nrf2 (nuclear factor erythrocyte-associated factor 2) is a transcription factor that regulates the expression of antioxidant proteins, enzymes, and other protective molecules in the body. Under normal conditions, Keap1 (Kelch-like ECH-associated protein 1) binds to Nrf2, rendering it inactive in the cytoplasm. The active components in the okra extract composition, such as quercetin 3-O-gentiobiglycoside (binding affinity -8.8) and isoquercetin (binding affinity -8.9), activate Nrf2 by interfering with the Keap1-Nrf2 interaction, leading to the release of Nrf2 and subsequent nuclear translocation. By influencing this pathway, quercetin and isoquercetin can help reduce oxidative stress, one of the main causes of fatigue and muscle damage during and after physical activity.

[0025] Quercetin 3-O-gentiobiose and isoquercetin, derived from okra extract compositions, modulate the activity of Silent Information Regulator 1 (SIRT1). SIRT1 is an NAD+-dependent deacetylase that plays a crucial role in regulating metabolism, cellular stress responses, and lifespan. The binding affinity of quercetin 3-O-gentiobiose from okra extract is -8.40, and that of isoquercetin from okra extract is -6.40. These binding affinities indicate that both compounds can interact effectively with SIRT1, although quercetin 3-O-gentiobiose may potentially activate SIRT1 more strongly than isoquercetin. By improving mitochondrial function, enhancing fatty acid oxidation, repairing muscle, and reducing oxidative stress, these flavonoids help the body adapt to physical activity, maintain energy levels, and recover more quickly after exercise.

[0026] Tumor necrosis factor-alpha (TNF-α) is a key pro-inflammatory cytokine primarily produced by macrophages and immune cells in response to infection, injury, or exercise-induced stress. During exercise, TNF-α levels may rise, especially after strenuous physical activity. This increase in TNF-α can lead to muscle soreness, delayed onset muscle soreness (DOMS), and generalized fatigue due to its effects on muscle repair, protein turnover, and inflammation. Excessive TNF-α can also impair muscle function by promoting the production of reactive oxygen species (ROS), leading to oxidative stress and muscle damage. Flavonoid glycosides from okra extract compositions regulate the production of cytokines, including TNF-α, by acting on various signaling pathways involved in inflammation. The binding affinity of quercetin 3-O-gentiobiose to TNF-α is -8.6 kcal / mol, and that of isoquercetin to TNF-α is -7.9 kcal / mol, indicating a strong binding interaction. The negative binding energies suggest that these two compounds may effectively bind to TNF-α, potentially modulating its activity and influencing the inflammatory process. The active components also reduce muscle inflammation, oxidative stress, and systemic fatigue, thereby enhancing athletic performance and faster recovery.

[0027] Mechanism of action of okra composition: This invention describes a composition derived from okra that enhances endurance and reduces fatigue by: Increase ATP levels in the blood, liver, and muscles Enhanced ATPase activity leads to higher energy production through dephosphorylation. Lowering blood creatine kinase (CK) levels Increase succinate dehydrogenase (SDH) enzyme levels Lowering lactate dehydrogenase (LDH) levels reduces lactate accumulation. Lowering serum urea nitrogen (SUN) levels Increase muscle glycogen levels, thereby improving endurance. Inhibition of adenosine A1 receptor Activation of adenosine A2A receptor Activation of peroxisome proliferator-activated receptor δ (PPARδ) Activation of AMP-activated protein kinase (AMPK) Inhibition of Kelch-like ECH-associated protein 1 (Keap1) activates nuclear factor erythroid 2-associated factor 2 (Nrf2). Activation of peroxisome proliferator activates receptor gamma coactivator 1α (PGC-1α). Enhance the activity of Silent Information Regulator 1 (SIRT1) Inhibit tumor necrosis factor-α (TNF-α) The okra-derived composition is a palatable, free-flowing powder administered in an effective dose. The composition contains polysaccharides, flavonoid glycosides, and carbohydrates to enhance endurance and reduce fatigue in subjects requiring improved endurance. As used herein, the term "subject" refers to the recipient of the administration. This may include vertebrates such as mammals, fish, birds, reptiles, or amphibians. Specifically, the term covers humans, non-human primates, horses, pigs, rabbits, dogs, sheep, goats, cattle, cats, guinea pigs, and rodents. It does not indicate any specific age or sex; therefore, unless otherwise stated, it includes both adults and newborns, regardless of sex.

[0028] As used herein, the terms "effective amount" and "effective dose" refer to an amount sufficient to produce the desired therapeutic effect or relieve undesirable symptoms while remaining below the threshold that would cause adverse side effects. Those skilled in the art will understand that effective doses can vary between individuals based on factors such as age, sex, disease state, genetic background, weight, extract formulation, availability of other active ingredients, and environmental influences.

[0029] The compositions of the present invention comprise 5-50% polysaccharides and 30-70% total carbohydrates. In one embodiment, the powder composition comprises 36.7% total carbohydrates and 11.5% polysaccharides.

[0030] The compositions of the present invention comprise 5-50% polysaccharides, 2-10% flavonoid glycosides, and 30-70% total carbohydrates. In one embodiment, the powder composition comprises 36.7% total carbohydrates, 5% flavonoid glycosides, and 11.5% polysaccharides.

[0031] The compositions of the present invention may also contain other components, such as one or more excipients.

[0032] In another aspect, the present invention provides the use of compositions containing okra extract to improve endurance or performance and reduce fatigue.

[0033] The compositions of the present invention can be formulated into various dosage forms. Non-limiting examples include capsules, tablets, pills, dispersions, suspensions, solutions, powders, tea, syrup concentrates, sticks, and chewable tablets.

[0034] The powder compositions of the present invention can also be used as compound ingredients in other products. Non-limiting examples include foods, pharmaceuticals, sports drinks, meal replacement drinks, powders, ointments, dietary supplements, capsules, tablets, or water additives.

[0035] In another embodiment, a method for enhancing the endurance of a subject and reducing the subject's fatigue includes administering an effective amount of the composition to a subject in need.

[0036] In one embodiment, the compositions disclosed herein are administered to a subject via oral administration.

[0037] In another embodiment, the composition is administered to mammals at a dose ranging from 100 mg / day to 2000 mg / day.

[0038] In one embodiment, administration of a composition of okra extract of 100 mg to 2000 mg to mammals over 30 days results in the following improvements.

[0039] 1. Improve running endurance 2. Enhance swimming endurance 3. Improve the subjective level of physical exertion. 4. Improves the severity of fatigue 5. Improve fatigue score 6. Increase peak power 7. Enhances endurance and muscle performance Preparation method The method for preparing the free-flowing powder composition involves: a) Powder the dried okra pods; b) Extract with an alcoholic solution at 90°C for 90 minutes (repeat three times); c) Collect and concentrate the filtrate at 70°C; d) Add ethyl acetate, stir, and allow to stand for separation; e) Remove ethyl acetate and concentrate the aqueous layer at 85°C; f) Add ethanol and cool at 4°C for 24 hours; and g) Clarify, centrifuge, and dry at 95°C to obtain a free-flowing powder.

[0040] Alternative methods use enzyme-assisted extraction, in which: a) Powder the dried okra pods and extract them with water at 90°C for 90 minutes (repeat three times). b) Combine all filtrates and heat at 60°C; c) Add pectinase and stir for 4 hours, and d) The solution was concentrated under vacuum at 95°C and then spray-dried.

[0041] The efficacy of the powdered composition was evaluated using a rat forced swim test (FST). In one embodiment, rats were administered a dose of okra extract at 100 mg / kg one hour prior to the swim test. The treated rats had an immobility time of 78 seconds, compared to 121 seconds in the normal control group.

[0042] Compared to the control group's immobility time of 121 seconds, rats treated with enzyme-treated okra extract showed an 81-second shorter immobility time. A shorter immobility time indicates better endurance and less fatigue. Furthermore, dried okra powder resulted in an immobility time of 115 seconds, comparable to the control group.

[0043] In another embodiment, the rat endurance test (FST) was used to assess the efficacy of the powdered composition. Rats administered okra extract at a dose of 100 mg / kg / day for 28 days showed a swimming time extension of 24 seconds compared to a swimming time of 13 seconds in the normal control group.

[0044] Rats administered the same dose of enzyme-treated okra extract also showed a 22-second longer swimming time. In contrast, rats administered only okra aqueous extract showed a swimming time of 16 seconds, while rats receiving dried okra powder swam for 14 seconds, comparable to the control group. Longer swimming time indicates better endurance and reduced fatigue.

[0045] In another implementation, the effect of okra extract on endurance in recreational runners was investigated. Subjects were divided into three groups: receiving 500 mg / day of okra extract for 30 days, receiving 1000 mg / day of okra extract for 30 days, or receiving a placebo for 30 days. The primary outcome measure was the mean change in total treadmill distance run in a 15-minute timed test from day 1 to day 30.

[0046] Based on data from a 15-minute timed test, participants who received 1000 mg / day of okra extract for 30 days showed the greatest improvement (62%) in running distance during the study period.

[0047] Participants who received 1000 mg / day of okra extract also showed the most significant improvement in perceived physical performance (RPE), with a 19% reduction during a 30-minute steady-state run and a 24% reduction during a 15-minute fast run, indicating better endurance and pacing strategies.

[0048] Significant improvements in fatigue severity were observed in subjects who received okra extract, with the most significant reduction in fatigue scores occurring over a 30-day period.

[0049] Following barbell lift tests, subjects who received okra extract for 30 days showed a decrease in fatigue scores. In the okra extract group, fatigue scores continued to decline over the 30-day period.

[0050] In addition, the peak power of the group receiving okra extract increased, indicating an overall improvement in jump performance over the study period.

[0051] Participants also showed positive changes in their ability to lift barbells overhead, demonstrating enhanced endurance and muscle performance after 30 days of okra extract administration.

[0052] Example A method for preparing okra extract.

[0053] 1 kg of powdered dried okra pods were extracted at 90°C with 15 L of water and methanol at a ratio of 80:20 for 90 minutes. After extraction, the first filtrate and the first residue were separated. The first residue was extracted again at 90°C with 15 L of water and methanol at a ratio of 80:20 for 150 minutes to form a second filtrate and a second residue. The second residue was extracted again at 90°C with 15 L of water and methanol at a ratio of 80:20 for 150 minutes to form a third filtrate and a third residue. The first, second, and third filtrates were combined and heated to 70°C to evaporate the methanol fraction, thus forming a concentrated aqueous fraction. 20 L of ethyl acetate was added to the concentrated aqueous fraction, stirred at room temperature for 2 minutes, and allowed to stand for 1 hour. After 1 hour, the aqueous layer and the ethyl acetate layer were separated. The washing process with ethyl acetate was repeated twice. The aqueous layer was separated from the ethyl acetate layer, and the aqueous layer was concentrated at 85°C until the TDS reached 20%. 100 liters of ethanol were slowly added to the concentrated aqueous solution, and the mixture was cooled at 4°C for 24 hours. After 24 hours, the mixture clarified, and the centrifuged fraction and residue were separated. The centrifuged fraction was concentrated under vacuum at 95°C until it reached a TDS concentration of 20%. The concentrated wash solution was spray-dried to obtain a free-flowing okra powder composition in 50% yield (Sample 1).

[0054] The effect of okra extract on swimming endurance was assessed using the forced swimming test (FST) in rats. Adult male Sprague-Dawley rats weighing 200-250 g were housed in polypropylene cages. The rats were housed at 22-24°C and 65% relative humidity, with alternating light and dark cycles every 12 hours. Animals were allowed free access to pelleted feed and filtered water. Animals were allowed to acclimatize to the environment for one week prior to the experiment. One hour before the swimming test, the rats were orally administered 100 mg / kg of the extract. The animals were divided into 6 groups of 5 rats each. The test consisted of exposing the animals to a water tank (70 cm high; 40 cm in diameter, containing water at 25°C and 50 cm high) for 5 minutes. During the initial minutes, each animal attempted to leave the water, then succumbed to the experimental conditions and assumed a typical immobile posture (defined as when no additional activity was observed other than the activity required to keep the head above water), occasionally attempting to escape. The total immobility duration during the test was recorded. In this test, shorter immobility time indicated better endurance / less fatigue.

[0055] Prolonged immobility is often interpreted as a marker of depressive-like behavior in rodents. Conversely, shortened immobility is considered a potential indicator of antidepressant effects. Endurance refers to the ability to sustain physical activity for an extended period. Fatigue refers to a decline in physical and mental performance following prolonged exertion. In forced swimming tests, rats with better endurance will be able to swim and remain active for longer periods before showing signs of fatigue. In forced swimming tests, shorter immobility times can be associated with better endurance. This is because shorter immobility suggests that the animal is more actively attempting to escape or cope with the stress of being placed in the water. Increased activity such as swimming and struggling indicates a higher level of endurance or adaptability in the face of challenging environments.

[0056] Compared to the normal control group, rats treated with okra aqueous extract exhibited a stillness time of 93 seconds. Group 6 rats treated with Sample 1 prepared according to Example 1 showed a stillness time of 78 seconds, compared to the normal control group which showed a stillness time of 121 seconds. Administration of 100 mg / kg of enzyme-treated okra extract to rats (Group 5) one hour before the swimming test resulted in a shortened stillness time of 81 seconds, compared to the normal control group which showed a stillness time of 121 seconds. The group treated with dried okra powder showed a stillness time of 115 seconds, comparable to the normal control group. Shorter stillness times indicate better endurance and less fatigue.

[0057] The effect of okra extract on swimming endurance was assessed using a rat (loaded) forced swimming test (FST). Adult male Sprague-Dawley rats weighing 200-250 g were confined in polypropylene cages. The rats were housed at 22-24°C and 65% relative humidity, with alternating light and dark cycles every 12 hours. Animals were allowed free access to pelleted feed and filtered water. The animals were allowed to acclimatize to the environment for one week before the experiment. The rats were divided into 6 groups of 5 rats each. The extract was orally administered to the rats at 100 mg / kg / day for 28 days. Thirty minutes after the last dose, the rats were removed from their groups for a weighted swimming test. Thirty minutes after the last gavage administration, the rats were placed individually in a swimming pool (70 cm deep; 40 cm in diameter, containing water up to 50 cm high at 25°C) in which the rats could not support themselves by touching the bottom with their tails. The rats were then loaded with a metal weight attached to their tails (approximately 5% of their body weight). The time to exhaustion was used as an indicator of forced swimming ability. A longer swimming time indicated better endurance / less fatigue.

[0058] The weighted forced swimming test was designed to impose a greater physical challenge on the animals, simulating conditions that could induce fatigue more quickly. Rats exhibiting prolonged swimming times under these challenging conditions were considered to have better endurance and a higher fatigue threshold. Group 6 animals treated for 28 days with okra extract prepared according to Example 1 at a dose of 100 mg / kg / day showed a longer swimming time of 24 seconds, compared to the normal control group which showed a swimming time of 13 seconds. Administration of enzyme-treated okra extract to rats at a dose of 100 mg / kg / day for 28 days also showed a longer swimming time of 22 seconds. After administration of okra aqueous extract alone, rats showed a swimming time of 16 seconds. The group treated with dried okra powder showed a swimming time of 14 seconds, with immobility times comparable to the normal control group. Longer swimming times indicate better endurance and less fatigue.

[0059] To determine the effect of okra extract on the endurance of recreational runners.

[0060] For this endurance study, male and female recreational active runners aged 25-40 years were selected. Participants were divided into three groups of 15 participants each (male:female ratio 1:1). The study sample was administered to the participants as follows.

[0061] Group 1: Subjects were given 500 mg of okra extract daily for 30 days (Sample 1).

[0062] Group 2: Subjects were given 1000 mg of okra extract (Sample 1) daily for 30 days.

[0063] Group 3: Subjects were given 1000 mg of placebo daily for 30 days.

[0064] The primary outcome measure was the average change in total distance traveled on the treadmill during a 15-minute timed test from day 1 to day 30.

[0065] Secondary outcome measures are • Average variation in peak power output during vertical jump testing from day 1 to day 30.

[0066] • The average change in time taken to reach exhaustion in barbell lifts and hold tests from day 1 to day 30.

[0067] • Mean change in the Borg 6-20 RPE scale from day 1 to day 30.

[0068] • Average change in FSS & VAS-F average scores from day 1 to day 30.

[0069] • Average change in biomarkers from day 1 to day 30.

[0070] Baseline primary and secondary outcome measures were performed before administering the stated dose to subjects. Following baseline measurements, subjects in each group were administered either the extract or placebo for 30 days. By day 30, all primary and secondary outcome measures had been completed.

[0071] 15-minute timed test Participants run for 15 minutes at their fastest pace that they can manage without becoming overly fatigued. The total distance run within 15 minutes is recorded. Participants choose their own pace.

[0072] Based on data from the 15-minute timed trial, participants' performance varied across groups. On Day 1, Group 1 ran 1.92 km, increasing to 2.81 km after 30 days of treatment (a 46% improvement). Group 2 started at 2.18 km on Day 1 and reached 3.53 km by the end (Day 30) (a 62% improvement). Although Group 3 started at 2.02 km on Day 1, it finished at 2.30 km by Day 30. The results indicate that participants in Group 2 achieved the greatest distance improvement over the 30-day period.

[0073] Subjective Physical Performance Rating (BORG) The BORG 6-20 Subjective Feeling Physical Exertion (RPE) scale is a 15-point scale ranging from 6 to 20, where 6 represents no exertion and 20 represents maximum exertion. RPE is captured every 5 minutes during exercise. The lower the RPE while maintaining a running heart rate of 80% of maximum heart rate, the better the performance.

[0074] Data showed that Group 2 exhibited the most significant improvement in RPE during steady-state 30-minute runs (19% decrease in perceived strength) and 15-minute fast runs (24% decrease in perceived strength), indicating a better endurance or pacing strategy. Group 1 showed moderate improvement in both steady-state runs and fast runs, indicating good performance, but the changes were less dramatic compared to Group 2.

[0075] In summary, lower RPE scores were associated with better performance because they indicated that participants were able to maintain their target heart rate with less effort. Group 2 performed best overall in terms of reducing perceived physical exertion.

[0076] Fatigue severity scale (FSS) The FSS questionnaire contains nine statements that assess the severity of your fatigue symptoms.

[0077] On day 1, the mean FSS scores for all three groups were relatively high, ranging from 36.1 to 35.4. This indicates that participants in all groups began experiencing moderate to severe fatigue symptoms. After 30 days (day 30), FSS scores decreased in all groups, indicating a reduction in fatigue severity. The data show a general improvement in fatigue severity across all groups over the 30-day period, with group 2 showing the most significant reduction (35%). Group 1 also showed a 20% reduction in fatigue scores. These findings suggest that intervention or treatment may have a positive effect on reducing fatigue symptoms. However, group 3 showed a relatively small change in fatigue severity (4%).

[0078] VAS-Fatigue (VAS-F) The Visual Analogue Scale for Fatigue uses a 10 cm scale, with one end representing no fatigue and the other end representing extreme fatigue.

[0079] On days 1 and 30, all groups (Groups 1, 2, and 3) had a pre-exercise fatigue score of 0, indicating no fatigue at the start of each assessment period. After performing barbell lifts, all groups showed similar fatigue scores on day 1. After 30 days of okra extract administration, fatigue scores decreased in both Group 1 (22% decrease) and Group 2 (37% decrease). Fatigue levels were quite similar across groups after the 15-minute timed test on day 1. After 30 days, fatigue scores decreased in both Group 1 (41% decrease) and Group 2 (55% decrease).

[0080] These findings suggest that okra extract can effectively reduce fatigue levels over a 30-day period, with greater fatigue reduction observed over time, particularly in group 2.

[0081] Vertical jump The vertical jump test is a jump test in which participants jump as high as possible from a squatting position, touching a vertical jump tester with one hand to determine the jump height. Three attempts are given, and the best of the three is recorded as the maximum height achieved. Peak power can then be calculated based on the jump height.

[0082] Peak power (W) = 60.7 × (jump height [cm]) + 45.3 × (body weight [kg]) - 2055.

[0083] On day 1 (D1), the peak power values ​​were quite similar across all groups. On day 30, the peak power values ​​of groups 1 and 2 increased, indicating an overall improvement in jump performance over the 30-day okra extract application period.

[0084] Data shows that, over a 30-day period, both Group 1 and Group 2 showed overall improvement in maximum jump height and peak power. While Group 2 exhibited the most significant improvement in peak power (7%), Group 1 also showed a significant improvement in peak power (4%).

[0085] Barbell lift and hold Participants lift a barbell (without weights) overhead and hold the position for as long as possible. Time to exhaustion is measured in minutes.

[0086] Both Group 1 and Group 2 showed significant improvements in endurance, with Group 2 exhibiting the greatest increase in hold time (19%). Group 1 also showed a 12% improvement in hold time. Overall, the data indicate that participants experienced positive changes in their ability to hold a barbell overhead during the 30-day period of okra extract administration.

[0087] in conclusion Extensive research and experiments have proven that this invention establishes the source of okra ( Abelmoschus esculentus This palatable, free-flowing powder composition effectively enhances endurance and reduces fatigue. The unique bioactive components of this composition, including quercetin 3-O-gentiobiglycoside, isoquercetin, rhamnose galacturon-glycan, and arabinogalactan, have been scientifically validated to regulate key metabolic pathways such as AMPK activation, ATP regulation, adenosine receptor regulation, and oxidative stress reduction, resulting in improved energy metabolism, muscle glycogen preservation, and delayed fatigue onset. Experimental data from animal models and human trials confirm significant improvements in physical endurance, sense of exertion, fatigue reduction, and muscle recovery, demonstrating the efficacy of this formulation. Therefore, this invention presents a novel, scientifically based, nutritional supplement-based method for endurance enhancement, providing a natural, safe, and effective solution for individuals seeking to improve endurance, athletic performance, and sustained energy levels.

Claims

1. A type derived from okra ( Abelmoschus esculentus A composition for enhancing endurance and reducing fatigue, the composition comprising: a) Polysaccharides, wherein the amount of said polysaccharides is in the range of 5% to 50%; b) Flavonoid glycosides, wherein the amount of said flavonoid glycosides is in the range of 2% to 10%, and c) Carbohydrates, wherein the amount of carbohydrates is in the range of 30% to 70%.

2. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition comprises: a) A polysaccharide, wherein the amount of the polysaccharide is in the range of 5% to 50%, wherein at least a portion comprises rhamnose-galacturonic acid polysaccharide and arabinogalactan; b) Flavonoid glycosides, wherein the amount of said flavonoid glycosides is in the range of 2% to 10%, wherein at least a portion comprises quercetin 3-O-gentiobiglycoside and isoquercetin (okra flavonoid glycosides), and c) Carbohydrates, wherein the amount of carbohydrates is in the range of 30% to 70%.

3. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition comprises: a) 36.7% total carbohydrates; b) 5% flavonoids, and c) 11.5% polysaccharides.

4. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition comprises excipients to enhance stability and bioavailability.

5. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition is in the form of a palatable, free-flowing, light brown, water-soluble powder.

6. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition is formulated into one or more dosage forms selected from: capsules, tablets, powders, tea, syrups, dispersions, solutions, sticks, chewable tablets, sports drinks, or dietary supplements.

7. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition is used as a compound ingredient in other foods, medicines, sports drinks, meal replacement drinks, ointments or water additives.

8. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition is administered orally to a subject.

9. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the effective dose of the composition is in the range of 100 mg / day to 2000 mg / day.

10. The okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, wherein the composition... a) Improve running endurance; b) Improve swimming endurance; c) Increase the subjective physical perception level (RPE); d) Improve the severity of fatigue; e) Improve fatigue score; f) Increase peak power; and g) Enhance endurance and muscle performance.

11. A method for preparing the okra-derived composition for enhancing endurance and reducing fatigue according to claim 1, the method comprising the following steps: a) Powder the dried okra pods; b) Extract the powder with a hydroethanol solution at 90°C for 90 minutes; c) Repeat the above process twice more; d) Combine all filtrates and concentrate at 70°C; e) Add ethyl acetate, stir for 2 minutes at room temperature, and let stand for 1 hour; f) Separate the ethyl acetate layer and the aqueous layer; g) Concentrate the water layer at 85°C; h) Add ethanol and cool the solution to 4°C for 24 hours; i) Clarify the solution, centrifuge, and collect the centrifuged mixture; as well as j) The centrifuged fraction is concentrated at 95°C and dried to obtain a free-flowing powder.

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