A composition for enhancing endurance derived from OKRA and a method of preparation and application

Abstract

The present invention relates to a palatable, free-flowing powdered composition derived from okra (Abelmoschus esculentus) for enhancing endurance. The composition comprises polysaccharides (5-50%), including Rhamnogalacturonan and Arabinogalactan; flavonoid glycosides (2-10%), including Quercetin 3-O-gentiobioside and Isoquercetin (Okraendurosides); and total carbohydrates (30-70%), with optional excipients for stability and bioavailability. The composition on administration to subjects in the appropriate dosage improves running endurance, enhances swimming endurance, improves rate of perceived Exertion (RPE), improves fatigue severity, improves fatigue score, increases peak power and enhances endurance and muscle performance. The invention also discloses a method of preparation involving hydroalcoholic extraction, concentration, and purification, resulting in a water-soluble powder. The composition can be formulated into capsules, tablets, powders, and beverages and is administered orally in doses of 100-2000 mg per day. Animal and human studies confirm its efficacy in enhancing endurance, making it applicable in nutraceutical, sports nutrition, and pharmaceutical industries.

Description

A COMPOSITION FOR ENHANCING ENDURANCE DERIVED FROM OKRA AND A METHOD OF PREPARATION AND APPLICATIONFIELD OF INVENTION

[0001] The present invention is related to a composition derived from Okra plant for enhancing endurance and reducing fatigue in subjects. The present invention also discloses a method of preparation of the composition and its application for endurance.BACKGROUND

[0002] Okra, scientifically known as Abelmoschus esculentus, belonging to the family Malvaceae, is commonly known as Lady’s finger, as well as by several vernacular names, including okra, bhindi, okura, quimgombo, bamia, gombo, and lai long ma, in the different geographical regions of its cultivation. The immature green pods of okra are widely consumed as vegetables, while the extract of the pods also serves as a thickening agent in numerous recipes for soups, as well as sauces, to augment their viscosity. Generally, okra is a high-value crop because it represents a source of nutrients that are important to human health, e.g., vitamins, potassium, calcium, carbohydrates, dietary fiber, and unsaturated fatty acids such as linolenic and oleic acids. Durazzo et al discloses that extracts of young okra pods have also been reported to display moisturizing and diuretic properties, whereas the seeds of this plant have been reported to possess anticancer and fungicidal properties. Durazzo et al evaluated the bioactive components and beneficial properties of Abelmoschus esculentus (Okra) like antidiabetic activity. Dantas et al discloses the extraction of Okra mucilage and its application as thickeners and food stabilizers. Xia Fangbo et al discloses the antioxidant and anti-fatigue constituents from Okra seeds and skins. Yan Li discloses a composition and antifatigue activity of water-soluble okra from Okra stem pectins.PRIOR ART

[0003] A review of prior art indicates that while okra (Abelmoschus esculentus) has been studied for anti-fatigue effects, there is limited evidence of patents specifically claiming a palatable powder composition derived from okra for endurance enhancement.

[0004] Notably, existing studies identify antioxidant and anti-fatigue properties in okra seeds and skins, suggesting their potential to reduce fatigue and improve endurance. However, they do not disclose a specific palatable powder formulation or a detailed method of preparationaimed at enhancing endurance. Patent application No. CN104223271A discloses an okra lactic acid bacteria fermented beverage, but it does not focus on endurance enhancement.

[0005] The present invention discloses a novel palatable powder composition derived from Okra specifically designed for enhancing endurance and reducing fatigue. This composition, along with its method of preparation and application, differentiates it from existing literature and patents.SUMMARY OF THE INVENTION

[0006] The present invention relates to a novel, palatable, free-flowing powdered composition derived from okra (Abelmoschus esculentus) for enhancing endurance and reducing fatigue in subjects. The invention also provides a method of preparation and application of this composition. The composition comprises polysaccharides, flavonoid glycosides, and carbohydrates and is formulated to improve endurance and reduce fatigue when administered in an effective amount.

[0007] The key bioactive components in the composition include Quercetin 3-O- gentiobioside and Isoquercetin (Okraendurosides) as flavonoid glycosides, and Rhamnogalacturonan and Arabinogalactan as polysaccharides. These components play a crucial role in modulating adenosine receptors (Al and A2A), AMP-activated protein kinase (AMPK), Peroxisome Proliferator-Activated Receptor Delta (PPAR5), and Nuclear factor erythroid 2-related factor 2 (Nrf2), thereby optimizing energy metabolism, reducing oxidative stress, and promoting sustained performance.

[0008] The composition for enhancing endurance and reducing fatigue derived from okra (Abelmoschus esculentus), according to the invention comprises of a) Polysaccharides in an amount ranging from 5% to 50%; b) Flavonoid glycosides in an amount ranging from 2% to 10%, and c) Carbohydrates in an amount ranging from 30% to 70%. In an embodiment under the invention, said composition contains: a) Polysaccharides in an amount ranging from 5% to 50%, wherein at least a portion including Rhamnogalacturonan and Arabinogalactan; b) Flavonoid glycosides in an amount ranging from 2% to 10%, wherein at least a portion including Quercetin 3-O-gentiobioside and Isoquercetin (Okraendurosides) and c) Carbohydrates in an amount ranging from 30% to 70%. In preferred embodiment under the invention, said composition contains: a) 36.7% total carbohydrates; b) 5% flavonoid glycosides and c) 11.5% polysaccharides. Excipients may be added to the composition to enhance stability and bioavailability. The composition is a palatable, free-flowing, lightbrown, water-soluble powder that can be formulated into various dosage forms, including capsules, tablets, powders, teas, syrups, dispersions, solutions, bars, chews, sports beverages, and dietary supplements. Additionally, the composition can be used as a compound ingredient in other food, medicine, sports beverages, meal replacement drinks, ointments, or water additives. It is intended for oral administration to subjects, with an effective dose ranging from 100 mg to 2000 mg per day to achieve the desired endurance-enhancing benefits.

[0009] Administering Okra extract composition as explained above to a mammal results in a) improved running endurance; b) enhanced swimming endurance; c) improved rate of perceived Exertion (RPE); d) improved fatigue severity e) improved fatigue score f) increased peak power and g) enhanced endurance and muscle performance.

[0010] The method of preparation involves hydroalcoholic 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 of preparing the composition for enhancing endurance derived from okra according to the invention comprises the steps of: a) powdering dried okra pods; b) extracting the powder with a hydroalcoholic solution at 90°C for 90 minutes; c) repeating the process two more times; d) combining all filtrates and concentrating at 70°C;e) adding ethyl acetate, stirring for 2 minutes at room temperature, and allowing to settle for 1 hour; f) separating the ethyl acetate layer and aqueous layer; g) concentrating the aqueous layer at 85°C; h) adding ethanol, cooling the solution to 4°C for 24 hours; i) clarifying the solution, centrifuging, and collecting the centrifugate; and j) concentrating the centrifugate at 95°C and drying to obtain a free-flowing powder.

[0012] The invention offers a novel nutraceutical and sports nutrition solution that provides sustained energy, improves performance, and accelerates recovery, making it beneficial for athletes, fitness enthusiasts, and individuals requiring enhanced stamina and endurance.DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention discloses a palatable, free flowing powder composition prepared from Okra (Abelmoschus esculentus). The powder composition comprises polysaccharides, flavonoid glycosides and carbohydrates. The invention also discloses the capability of the composition for enhancing endurance and reducing fatigue to the subjectwhen administered in an effective amount of the composition and method of preparation of said palatable powder composition.

[0014] The powder composition of the present invention comprises Quercetin 3-0- gentiobioside and isoquercetin (collectively known as Okraendurosides) as flavonoid glycosides. It also comprises Rhamnogalacturonan and Arabinogalactan as polysaccharides.

[0015] Okra, (Abelmoschus esculentus) 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 folate. One of the distinctive features of okra is the mucilage (slimy substance), which is released when the pods are cut or cooked. The mucilage is having thickening properties and is commonly used in some traditional dishes. However, the mucilaginous nature of okra mucilage may cause digestive discomfort in some individuals, leading to bloating or gas. In the present invention, a palatable, free- flowing, light brown powder composition, soluble in water, is obtained from okra and used to enhance the endurance.

[0016] Role of ATP in Endurance Enhancement: Endurance refers to the body's ability to sustain prolonged physical activity or resist fatigue over an extended period. It involves the cardiovascular, respiratory, musculoskeletal, and metabolic systems, which must work together to support prolonged or repetitive exercise. Endurance activities place significant demand on the Adenosine Triphosphate (ATP) energy system, requiring the body to efficiently produce and regenerate ATP, which is essential for sustained performance.

[0017] ATP is the primary energy currency in cells. The energy stored in ATP is used for various cellular processes, such as: Muscle contraction; Active transport of ions across cell membranes and Synthesis of macromolecules like 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. The enzyme ATPase (Adenosine Triphosphatase) catalyzes the hydrolysis of ATP to ADP (Adenosine Diphosphate) and inorganic phosphate (Pi), releasing energy that cells use for muscle contraction and other biological functions.

[0019] The Adenosine Al receptor protein plays a significant role in regulating energy metabolism, fatigue, and endurance during physical activity. Activation of Al receptors lead to reduced cyclic AMP (cAMP) levels, which has a dampening effect on cellular activity,promoting fatigue and reducing exercise performance. Quercetin 3-O-gentiobioside and isoquercetin present in the Okra extract composition could potentially bind to the Al receptor, blocking adenosine’s fatigue -inducing effects or modulating receptor activity, thereby reducing the perception of fatigue and enhancing endurance. The binding affinity of Quercetin 3-O-gentiobioside is -10.2 kcal / mol and isoquercetin is -8.7 kcal / mol. This lower binding affinity suggests a strong interaction between Quercetin 3-O-gentiobioside and the Adenosine Al receptor, which could lead to enhanced endurance and reduced fatigue.

[0020] Adenosine A2A receptor also plays a key role in regulating energy metabolism, inflammation, and muscle function. Quercetin 3-O-gentiobioside and isoquercetin from Okra extract of the present invention binds and activate the Adenosine A2A receptor and thereby enhance the endurance and reduce fatigue during physical exertion. The binding affinity of Quercetin 3-O-gentiobioside is -8.40 kcal / mol and isoquercetin is -7.40 kcal / mol indicating that both compounds have strong binding interactions with the Adenosine A2A receptor. A more negative binding affinity value indicates a stronger and more stable interaction between a ligand (the compound) and its receptor. By activating the Adenosine A2A receptor, Quercetin 3-O-gentiobioside and isoquercetin may promote better energy balance and metabolism during exercise, helping the body conserve energy and delay the onset of fatigue.

[0021] Quercetin 3-O-gentiobioside and isoquercetin from Okra extract composition improve Peroxisome Proliferator-Activated Receptor Delta (PPAR5), a key regulator of lipid metabolism, fatty acid oxidation, and muscle endurance. The binding affinity of -9.0 kcal / mol for Quercetin 3-O-gentiobioside and -7.9 kcal / mol for isoquercetin suggest relatively strong interactions between both compounds and PPAR5. PPAR5 activation encourages the use of fatty acids as the primary energy source, preserving glycogen stores and providing a sustained energy supply during prolonged physical exertion.

[0022] AMP-activated protein kinase (AMPK) is a crucial enzyme in the regulation of energy balance, and its activation can enhance endurance, reduce fatigue, and improve overall physical performance. Quercetin 3-O-gentiobioside and isoquercetin present in the Okra extract composition activates AMPK through multiple mechanisms (increasing the AMP / ATP ratio, reducing oxidative stress). The binding affinity of -9.6 kcal / mol for Quercetin 3-O- gentiobioside and -9.2 kcal / mol for isoquercetin suggest that both compounds bind relatively strongly to AMPK. A negative binding energy value in this range indicates a stable and favourable interaction between the compounds and AMPK, meaning these compounds caneffectively activate the receptor. AMPK activation increases energy production, enhances fat oxidation, and promotes mitochondrial biogenesis, all of which support improved endurance during prolonged physical activity. By optimizing energy use, promoting efficient recovery, and reducing oxidative stress, quercetin and isoquercetin help delay fatigue and improve overall exercise performance.

[0023] Peroxisome proliferator-activated receptor gamma coactivator 1 -alpha (PGC-la ) plays a critical role in enhancing endurance and reducing fatigue by improving the body’s ability to generate and utilize energy efficiently. Quercetin 3-O-gentiobioside and isoquercetin present in the Okra extract composition significantly enhance endurance and reduce fatigue by activating PGC-la. The binding affinity of -8 kcal / mol for both Quercetin 3-O-gentiobioside and isoquercetin suggests that both compounds exhibit moderate to strong binding to PGC-la. PGC-la activation promotes more efficient energy production and better mitochondrial function, muscles are able to perform for longer periods before reaching fatigue. PGC-la activation contributes to reducing inflammation, which helps to mitigate the soreness and discomfort that typically accompany intense physical exertion, further reducing fatigue during exercise.

[0024] Nrf2 (Nuclear factor erythroid 2-related factor 2) is a transcription factor that regulates the expression of antioxidant proteins, enzymes, and other protective molecules in the body. Under normal conditions, Keapl (Kelch-like ECH-associated protein 1) binds to Nrf2, keeping it inactive in the cytoplasm. The active components in the Okra extract composition like Quercetin 3-O-gentiobioside (binding affinity-8.8) and isoquercetin (binding affinity-8.9) activate Nrf2 by interfering with the Keapl -Nrf2 interaction, leading to the release and subsequent nuclear translocation ofNrf2. By affecting this pathway, quercetin and isoquercetin can help reduce oxidative stress, which is one of the primary contributors to fatigue and muscle damage during and after physical activity.

[0025] Quercetin 3-O-gentiobioside and isoquercetin from Okra extract composition modulate sirtuin 1 (SIRT1) activity. SIRT1 is a NAD+-dependent deacetylase that plays a crucial role in regulating metabolism, cellular stress response, and longevity. The binding affinity of Quercetin 3-O-gentiobioside is -8.40 and isoquercetin from Okra extract is -6.40. These binding affinities suggest that both compounds can interact effectively with SIRT1, although Quercetin 3-O-gentiobioside may potentially activate SIRT1 more strongly than isoquercetin. Through improved mitochondrial function, enhanced fatty acid oxidation,muscle repair, and reduced oxidative stress, these flavonoids help the body adapt to physical exertion, sustain energy levels, and recover more quickly after exercise.

[0026] Tumor Necrosis Factor-alpha (TNF-a) is a key pro-inflammatory cytokine produced primarily by macrophages and immune cells in response to infection, injury, or exercise- induced stress. During exercise, TNF-a levels can rise, particularly after intense physical activity. This increase in TNF-a can contribute to muscle soreness, delayed onset muscle soreness (DOMS), and a general feeling of fatigue due to its effects on muscle repair, protein turnover, and inflammation. Excess TNF-a 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 composition modulate cytokine production, including TNF-a, by acting on various signaling pathways involved in inflammation. The binding affinities of -8.6 kcal / mol for Quercetin 3-O-gentiobioside and -7.9 kcal / mol for isoquercetin suggest strong binding interactions with TNF-a. The negative binding energies indicate that both compounds are likely to bind effectively to TNF-a, potentially modulating its activity and influencing inflammatory processes. The active components also reduce s muscle inflammation, oxidative stress, and systemic fatigue, thereby enhancing better exercise performance and faster recovery.Mechanism of Action of the Okra Composition:

[0027] The present invention describes a composition derived from okra that enhances endurance and reduce fatigue by:• Increasing ATP levels in blood, liver, and muscles• Enhancing ATPase activity, leading to higher energy production through dephosphorylation• Decreasing creatine kinase (CK) levels in blood• Increasing succinate dehydrogenase (SDH) enzyme levels• Reducing lactate dehydrogenase (LDH) levels, lowering lactic acid accumulation• Decreasing serum urea nitrogen (SUN) levels• Enhancing muscle glycogen levels, improving endurance• Inhibiting Adenosine Al receptor• Activating Adenosine A2A receptor• Activating Peroxisome Proliferator- Activated Receptor Delta (PPAR5)• Activating AMP-activated protein kinase (AMPK)• Inhibit Kelch-like ECH-associated protein 1 (Keapl) thereby activate Nuclear factor erythroid 2-related factor 2 (Nrf2)• Activating Peroxisome proliferator-activated receptor gamma coactivator 1 -alpha (PGC- la )• Enhance sirtuin 1 (SIRT1) activity• Inhibit Tumor Necrosis Factor-alpha (TNF-a)

[0028] The okra-derived composition is a palatable, free-flowing powder, administered in an effective dosage form. The composition contains polysaccharides, flavonoid glycosides, and carbohydrates to enhance endurance and reduce fatigue in subjects requiring improved stamina. As used herein, the term "subject" refers to the recipient of administration. This may include vertebrates, such as mammals, fish, birds, reptiles, or amphibians. The term specifically encompasses humans, non-human primates, horses, pigs, rabbits, dogs, sheep, goats, cows, cats, guinea pigs, and rodents. It does not indicate any specific age or sex; therefore, both adults and newborns, whether male or female, are included unless stated otherwise.

[0029] As used herein, the terms "effective amount" and "effective dose" refer to a quantity sufficient to produce the desired therapeutic effect or to alleviate undesirable symptoms, while remaining below the threshold for causing adverse side effects. A person skilled in the art understands that the effective amount may vary between individuals based on factors such as age, sex, disease state, genetic background, body weight, formulation of the extract, availability of other active ingredients, and environmental influences.

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

[0031] The composition of the present invention comprises 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.

[0032] Compositions of the present invention may also comprise other components, for example, one or more excipients.

[0033] In another aspect, the present invention provides use of the composition comprising extract of Okra that improves endurance or performance and reduce fatigue.

[0034] The composition of the present invention can be formulated into various dosage forms. Non-limiting examples include capsules, tablets, pills, dispersions, suspensions, solutions, powders, teas, syrup concentrates, bars, and chews.

[0035] The powder composition of the invention may also be used as a compound ingredient in other products. Non-limiting examples include food, medicine, sports beverages, meal replacement drinks, powders, ointments, dietary supplements, capsules, tablets, or a water additive.

[0036] In another embodiment, a method for enhancing endurance and reducing fatigue in a subject comprises administering an effective amount of the composition to a subject in need thereof.

[0037] In one embodiment, the composition disclosed herein is administered to a subject via the oral route.

[0038] In another embodiment, the composition is administered to a mammal at a dose ranging from 100 to 2000 mg per day.

[0039] In one embodiment administering 100 mg to 2000mg of Okra extract composition to a mammal result in the following improvement over 30 days.1. Improvement in the running endurance2. Enhancing swimming endurance3. Improvement in rate of perceived Exertion4. Improvement in fatigue severity5. Improvement in fatigue score6. Improvement in peak power7. Enhancement in endurance and muscle performanceMethod of Preparation

[0040] The preparation method for the free-flowing powder composition involves: a) Powdering dried okra pods; b) Extracting with hydroalcoholic solution at 90°C for 90 minutes (repeated three times); c) Collecting and concentrating fdtrates at 70°C;d) Adding ethyl acetate, stirring, and allowing separation; e) Removing ethyl acetate and concentrating the aqueous layer at 85°C; f) Adding ethanol and cooling at 4°C for 24 hours; and g) Clarifying, centrifuging, and drying at 95 °C to obtain a free-flowing powder.

[0041] An alternative method uses enzyme-assisted extraction, where: a) Dried okra pods are powdered and extracted with water at 90°C for 90 minutes (repeated three times); b) combining all fdtrates and heated at 60°C; c) Pectinase enzyme is added and stirred for 4 hours and d) The solution is concentrated under vacuum at 95°C and spray-dried.

[0042] The efficacy of the powdered composition was evaluated using the forced swim test (FST) in rats. In one embodiment, rats were administered Okra extract at a dose of 100 mg / kg, one hour before the swimming test. The immobility time for the treated rats was 78 seconds, compared to 121 seconds in the normal control group.

[0043] Rats administered enzyme-treated Okra extract also showed a reduced immobility time of 81 seconds, compared to 121 seconds in the control group. A shorter immobility time indicates better endurance and reduced fatigue. Additionally, Okra dried powder resulted in an immobility time of 115 seconds, which was comparable to the control group.

[0044] In another embodiment, the efficacy of the powdered composition was evaluated using the weight-loaded forced swim test (FST) in rats to assess endurance capacity. Rats administered Okra extract at a dosage of 100 mg / kg / day for 28 days showed an increased swimming time of 24 seconds, compared to 13 seconds in the normal control group.

[0045] Rats administered enzyme -treated Okra extract at the same dosage also demonstrated an extended swimming time of 22 seconds. In contrast, rats administered Okra water extract alone showed a swimming time of 16 seconds, while those receiving Okra dried powder had a swimming time of 14 seconds, which was comparable to the control group. A longer swimming time indicates better endurance and reduced fatigue.

[0046] In another embodiment, the effect of Okra extract on endurance was studied in recreational runners. Subjects were divided into three groups, receiving either 500 mg / day of Okra extract for 30 days, 1000 mg / day of Okra extract for 30 days, or a placebo for 30 days,respectively. The primary outcome measure was the mean change in total treadmill distance covered in a 15-minute timed test, from Day 1 to Day 30.

[0047] Based on the 15 -minute time trial data, participants administered 1000 mg / day of Okra extract for 30 days showed the greatest improvement (62%) in distance covered over the study period.

[0048] Participants in the 1000 mg / day Okra extract group also demonstrated the most significant improvement in Rate of Perceived Exertion (RPE), with a 19% decrease in perceived exertion during the steady-state 30-minute run and a 24% reduction during the 15- minute fast run, indicating better endurance and pacing strategies.

[0049] A notable improvement in fatigue severity was observed in subjects administered Okra extract, with the most significant reduction in fatigue scores occurring over the 30-day period.

[0050] After performing the bar lift test, subjects who received Okra extract for 30 days showed a decrease in fatigue scores. The fatigue score continued to decrease over the 30-day period in groups administered Okra extract.

[0051] Additionally, peak power increased in groups that received Okra extract, suggesting a general improvement in jump performance over the study period.

[0052] Participants also showed a positive change in their ability to hold the barbell overhead, demonstrating enhanced endurance and muscle performance after 30 days of Okra extract administration.ExamplesMethod of preparation of Okra extract.

[0053] IKg powdered Okra dried pods were extracted with 15L of water and methanol in 80:20 ratio at 90°C for 90 minutes. After the extraction, first filtrate and first residue were separated. First residue was again extracted with 15L of water: methanol in 80:20 ratio at 90°C for 150 minutes to form second filtrate and second residue. Second residue was again extracted with 15L of water: methanol in 80:20 ratio at 90°C for 150 minutes to form third filtrate and third residue. First, second and third filtrates were combined and heated to 70°C to evaporate methanol part to form concentrated aqueous part. 20 Liters of ethyl acetate was added to the concentrated aqueous part, stirred for 2 minutes at room temperature and allowed to settle for 1 hour. After 1 hour, aqueous and ethyl acetate layers were separated.Washing process with ethyl acetate was repeated two more times. The aqueous layer was separated from ethyl acetate layer and the aqueous part was concentrated at 85°C until the TDS reached 20%. 100 liters of ethanol was added slowly to the concentrated aqueous solution and the mixture was cooled at 4°C for 24 hrs. After 24 hours, mixture was clarified and centrifugate and residue were separated. Centrifugate was concentered under vacuum at 95°C until it reached a TDS concentration of 20%. Concentrated wash was spray dried to obtain free flowing powder composition of Okra with 50% yield (sample 1).Evaluation of swimming endurance of Okra extracts by forced swim test (FST) in rats

[0054] Adult male albino rats (Sprague-Dawley) weighing 200- 250g were housed in polypropylene cages. The rats were kept at 22-24°C and 65% relative humidity with alternate light and dark periods of 12-h intervals each. Animals had free access to pellet food and filtered water. The animals were acclimatized for one week before the start of the experiment. Extracts were orally fed to rats at 100 mg / kg, 1 hour before swimming test. Animals were divided into 6 groups with 5 rats in each group. The test included exposure of the animals to a water tank (height, 70 cm; diameter, 40 cm, containing water up to 50 cm of height at 25°C) for 5 minutes duration. Each animal made vigorous attempts to get out of water during the first couple of minutes and thereafter surrendered to experimental conditions and assumed a typical immobile posture (which is defined as when no additional activity was observed other than that required to keep the head above the water) with occasional escape attempts. The total duration of immobility in the test session was recorded. In this test, less immobility time indicates better endurance / less fatigue.

[0055] Increased immobility time is often interpreted as a sign of depressive-like behavior in rodents. Conversely, a reduction in immobility time is considered a potential indicator of an antidepressant effect. Endurance is the ability to sustain physical activity over an extended period. Fatigue is the decline in physical and mental performance that occurs after prolongedexertion. In the context of the force swim test, a rat with better endurance would be able to swim and remain active for a longer duration before showing signs of fatigue. In the forced swim test, less immobility time can be associated with better endurance. This is because reduced immobility time implies that the animal is more actively attempting to escape or cope with the stressful situation of being placed in water. Increased activity, such as swimming and struggling, indicates a higher level of endurance or resilience in the face of a challenging environment.

[0055] Rats treated with Okra water extract showed an immobility time of 93 seconds compared to normal control group. Group 6 rats treated with sample 1 prepared as per example 1 showed less immobility time of 78 seconds compared to normal control group which showed an immobility time of 121 seconds. Administering Okra extract treated with enzyme at a dosage of lOOmg / Kg to rats (group 5)1 hour before swimming test showed less immobility time of 81 sec compared to normal control group which showed an immobility time of 121 seconds. Okra dried powder treated group showed an immobility time of 115 seconds which is comparable to normal control group. Less immobility time indicates better endurance and less fatigue.Evaluation of swimming endurance of Okra extracts by (weight-loaded) forced swim test (FST) in rats

[0056] Adult male albino rats (Sprague-Dawley) weighing 200- 250g were housed in polypropylene cages. The rats were kept at 22-24°C and 65% relative humidity with alternate light and dark periods of 12-h intervals each. Animals had free access to pellet food and fdtered water. The animals were acclimatized for one week before the start of the experiment. Rats were divided into 6 groups and each group contain 5 rats. Extracts were orally fed to rats at 100 mg / kg / day for 28 days. After 30 min of last dose, rats were taken out from each group for a weight-loaded swimming test. 30 min after the last intragastric administration, the rats were placed individually in a swimming pool (height, 70 cm; diameter, 40 cm, containing water up to 50 cm of height at 25°C), in which the rats could not support themselves by touching the bottom with their tails. The rats were loaded with a metal block (approximately 5% of body weight) attached to their tails. The swimming time to exhaustion was used as the index of the forced swimming capacity. More swimming time indicates better endurance / less fatigue.

[0058] The forced swim test with a weight load is designed to impose a greater physical challenge on the animals, simulating conditions that may induce fatigue more quickly. Rats that exhibit extended swimming times under these challenging conditions are considered to have better endurance and a higher threshold for fatigue. Group 6 animals treated with Okra extract prepared as per example 1 at a dosage of lOOmg / Kg / day for 28 days showed high swimming time of 24 seconds compared to normal control group which showed a swimming time of 13 secs. Administering Okra extract treated with enzyme at a dosage of lOOmg / Kg / day for 28 days to rats also showed high swimming time of 22seconds. After administering Okra water extract alone to rats showed a swimming time of 16 seconds. Okra dried powder treated group showed a swimming time of 14 seconds which is comparable to normal control group. A higher swimming time indicates better endurance and less fatigue.Determination the effect of Okra extract on Endurance in Recreational runners.

[0057] For this endurance study, 25-40 years old recreationally active runners of both genders were selected. Subjects were divided in to 3 groups with 15 subjects in each group (1: 1 male: female). Subjects were administered with study samples in the following manner.Group 1: Subjects administered with 500 mg of Okra extract (sample 1) per day for 30 days.Group 2: Subjects administered with 1000 mg of Okra extract (sample 1) per day for 30 daysGroup 3: Subjects administered with 1000 mg placebo per day for 30 days.

[0058] The primary outcome measure is the mean change in the total distance covered on treadmill in the 15 min timed teat from Day 1 to Day 30.

[0059] The secondary outcome measures areMean change in peak power output in the vertical jump test from Day 1 to Day 30.Mean change in time to exhaustion in the Barbell lift and hold test from Day 1 to Day 30.• Mean change in Borg 6-20 RPE scale from Day 1 to Day 30.• Mean change in the mean score of FSS & VAS-F from Day 1 to Day 30.• Mean change in biomarkers from Day 1 to Day 30.

[0060] Before administering the dosage to subjects, the baseline primary and secondary outcome measures were done. After baseline measurements, subjects in each group were administered with extract or placebo and continued for 30 days. At day 30, all primary and secondary outcome measures were done.15 min Time trial

[0061] The participant ran as fast as he could but without overexertion for 15 minutes. The total distance covered in 15 minutes was recorded. The speed was self-selected by the participant.

[0064] Based on the data from the 15-minute time trial, the participants' performance showed variation across different groups. On Day 1, Group 1 covered 1.92 km, increasing to 2.81 km after 30 days of treatment (46% improvement). Group 2 had a starting distance of 2. 18 km on Day 1, reaching 3.53 km by the end (D30) (62% improvement). While Group 3 began with 2.02 km on Day 1 and at Day 30, they finished with 2.30 km. The results suggested that participants in Group 2 achieved the greatest improvement in distance over the 30-day period.Rate of perceived Exertion (BORG)

[0062] The BORG 6-20 Rate of Perceived Exertion (RPE) scale is a 15 -point scale that ranged from 6 to 20, with 6 representing no exertion and 20 representing maximum exertion. The RPE was captured every 5 minutes during the exercise. The lower the RPE, while maintaining a running heart rate of 80% of max heart rate, the better the performance.

[0066] The data revealed that Group 2 showed the most significant improvement in RPE, in the steady-state 30-minute run (19% decrease in perceived exertion) and 15 minutes fat run (24% reduction in perceived exertion), which suggested better endurance or pacing strategies. Group 1 demonstrated modest improvements in both steady-state and fast runs, indicating good performance but less drastic changes compared to Group 2.

[0063] In summary, lower RPE scores were associated with better performance, as they indicated that participants were able to maintain the target heart rate while perceiving less effort. Group 2 displayed the best overall performance in terms of reduced perceived exertion.Fatigue severity scale (FSS)

[0064] The FSS questionnaire contained nine statements that rated the severity of your fatigue symptoms.

[0065] At DI, the average FSS scores for all three groups are relatively high, ranging from 36.1 to 35.4. This suggests that participants in all groups started with moderate to severe fatigue symptoms. After 30 days (D30), the FSS scores have decreased across all groups, indicating a reduction in fatigue severity. The data suggests that, over the 30-day period, there was a general improvement in fatigue severity across all groups, with Group 2 showing the most significant reduction (35% reduction). Group 1 also showed reduction in fatigue score by 20%. The findings indicate that the intervention or treatment may have had a positive impact on reducing fatigue symptoms. However, Group 3 showed a relatively small change (4%) in fatigue severity.VAS- Fatigue (VAS-F)

[0066] The Visual Analogue Scale for fatigue is a 10 cm scale with one end as No Fatigue and the other end as Extreme Fatigue.

[0071] At DI and D30, all groups (Group 1, Group 2, Group 3) had a fatigue rating of 0 before exercise, indicating no fatigue at the start of each assessment period. After performing the bar lift, all groups at DI showed similar fatigue score. After administering Okra extract for 30 days, the fatigue score decreased for Group 1 (22% decrease) and Group 2 (37% decrease). Fatigue levels after the 15 minutes time trial on Day 1 were fairly similar across groups. After 30 days, the fatigue score decreased for Group 1 (41% decrease) and group 2 (55% reduction).

[0067] These findings suggest that Okra extract may be effective in reducing fatigue levels over a 30-day period, with a greater reduction in fatigue observed as time progressed, particularly in Group 2.Vertical Jump

[0068] The vertical jump test is a jump test wherein the participant jumped as high as possible from a squatting position and touched a vertical jump tester with one hand to determine jump height. Three chances were given, and the best of the three was taken as the maximum height reached. Peak power could then be calculated from the jump height.Peak Power (W) = 60.7 x (jump height [cm]) + 45.3 x (body mass [kg]) - 2055.

[0074] On Day 1 (DI), peak power values were fairly similar across all groups. On Day 30, peak power increased in groups 1 and 2, indicating a general improvement in jump performance over the 30-day period of Okra extract administration.

[0069] The data shows an overall improvement in both maximum jump height and peak power over the 30-day period in group 1 and 2. While Group 2 showed the most significant improvement (7%) in peak power, Group 1 also showed notable improvements (4%).Barbell lift and hold

[0070] The participant lifted the barbell (without the weights) overhead and held the position as long as possible. The time to exhaustion was measured in minutes.

[0077] Group 1 and Group 2 showed a clear improvement in endurance, with Group 2 demonstrating the largest increase in hold time (19% increase in holding time). Group 1 also showed 12% improvement in holding time. Overall, the data suggests that the participants experienced a positive change in their ability to hold the barbell overhead over the 30-day period of Okra extract administration.Conclusion

[0071] Through extensive research and experimentation, the present invention establishes that a palatable, free-flowing powdered composition derived from okra (Abelmoschus esculentus) effectively enhances endurance and reduces fatigue. The composition's unique bioactive components, including Quercetin 3-O-gentiobioside, Isoquercetin, Rhamnogalacturonan, and Arabinogalactan, have been scientifically validated to modulate key metabolic pathways, such as AMPK activation, ATP regulation, Adenosine receptor modulation, and oxidative stress reduction, leading to improved energy metabolism, muscle glycogen preservation, and delayed fatigue onset. Experimental data from animal models and human trials confirm significant improvements in physical endurance, perceived exertion, fatigue reduction, and muscle recovery, demonstrating the efficacy of this formulation. The invention thus presents a novel, scientifically-backed, and nutraceutical-based approach to endurance enhancement, offering a natural, safe, and effective solution for individuals requiring improved stamina, athletic performance, and sustained energy levels.

Claims

We Claim:

1. A composition for enhancing endurance and reducing fatigue derived from okra (Abelmoschus esculentus), comprising: a) Polysaccharides in an amount ranging from 5% to 50%; b) Flavonoid glycosides in an amount ranging from 2% to 10%, and c) Carbohydrates in an amount ranging from 30% to 70%.

2. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1 wherein said composition contains: a) Polysaccharides in an amount ranging from 5% to 50%, wherein at least a portion including Rhamnogalacturonan and Arabinogalactan; b) Flavonoid glycosides in an amount ranging from 2% to 10%, wherein at least a portion including Quercetin 3-O-gentiobioside and Isoquercetin (Okraendurosides) and c) Carbohydrates in an amount ranging from 30% to 70%.

3. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1 wherein said composition contains: a) 36.7% total carbohydrates; b) 5% flavonoid glycosides and c) 11.5% polysaccharides.

4. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1 wherein said composition contains excipients to enhance stability and bioavailability.

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

6. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1 wherein said composition is formulated into one or more dosage forms, selected from Capsules, tablets, powders, teas, syrups, dispersions, solutions, bars, chews, sports beverages, or dietary supplements.

7. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1 wherein said composition is used as a compound ingredient in other food, medicine, sport beverages, meal replacement drinks, ointments, or water additives.

8. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1, wherein the composition is administered orally to subjects.

9. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1, wherein the effective dose of the composition ranges from 100 mg to 2000 mg per day.

10. The composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1, wherein said composition a) improves running endurance; b) enhances swimming endurance; cjimproves rate of perceived Exertion (RPE); d) improves fatigue severity; e) improves fatigue score; f) increases peak power and g) enhances endurance and muscle performance.

11. A method of preparing the composition for enhancing endurance and reducing fatigue derived from okra as claimed in claim 1, comprising the steps of: a) powdering dried okra pods; b) extracting the powder with a hydroalcoholic solution at 90°C for 90 minutes; cjrcpcating the process two more times; d) combining all filtrates and concentrating at 70°C; e) adding ethyl acetate, stirring for 2 minutes at room temperature, and allowing to settle for 1 hour; f) separating the ethyl acetate layer and aqueous layer; g) concentrating the aqueous layer at 85°C; h) adding ethanol, cooling the solution to 4°C for 24 hours; i) clarifying the solution, centrifuging, and collecting the centrifugate; and j) concentrating the centrifugate at 95°C and drying to obtain a free-flowing powder.

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