Compositions comprising punica granatum whole spectrum extract selectively enriched with punicagalins, ellagic acid and urolethin a and process for preparation thereof

Abstract

The present invention discloses compositions of Punica granatum whole spectrum extract (PWSE) extract comprising Urolithin A in the range of 0.2% - 10%, for improving general health, overall wellbeing and longevity by preventing age related disorders. The invention further discloses compositions comprising Punica granatum whole spectrum extract enriched with Punicagalins, Ellagic acid and Urolethin A. The invention additionally discloses a process for preparation of Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins, Ellagic acid and Urolethin A.

Description

[0001] Compositions comprising Punica granatum whole spectrum extract selectively enriched with Punicagalins, Ellagic acid and Urolethin A and process for preparation thereof

[0002] Technical filed:

[0003] The present invention relates to compositions of Punica granatum whole spectrum extract (PWSE) extract comprising Urolithin A in the range of 0.2% - 10%, for improving general health, overall wellbeing and longevity by preventing age related disorders. The invention further relates to compositions comprising Punica granatum whole spectrum extract selectively enriched with Punicagalins, Ellagic acid and Urolethin A. The invention further relates to process for preparation of Punica granatum whole spectrum extract selectively enriched with Punicagalins, Ellagic acid and Urolethin A.

[0004] Background and Prior Art:

[0005] It is well reported that the extracts of Punica granatum also known as pomegranate, contain a number of biologically active polyphenolic compounds; among these polyphenols, ellagitannins are of outstanding biological significance in view of their antioxidant, antimicrobial and radical scavenging activity.

[0006] It is well documented that the phytochemicals such as punicalagins present in the extracts of Punica granatum capable of preventing cardiovascular diseases (US8221806) by virtue of their powerful antioxidant and anti-inflammatory affects. These punicalagins exhibit cardioprotective activity by various mechanisms for example by inhibiting the oxidation of LDL, decreasing activity of angiotensin converting enzyme, increasing the production of nitric oxide (US8372454) and reducing oxidative stress and inflammatory damage in blood vessels, etc.

[0007] Studies have shown that the extracts of Punica granatum contains many phytochemicals, such as anthocyanins, ellagitannins, gallic acid, lignans along with other phytochemicals such as Gallotannins, Ferulic acid, Catechins, Quercetin, fatty acids, organic and phenolic antioxidants together produce synergistic effect enhancing healthy benefits of the extracts. Punicalagin is an ellagitannin has anti oxi dative, anti-inflammatory, hepatoprotective, antibacterial, and anticancer effects. However, ellagitannins having low bioavailability will be hydrolysed in the stomach to ellagic acid which in turn metabolized by the intestinal microbiome to urolithins prior to absorption. Thus, the ellagitannins are metabolized via ellagic acid by the intestinal microbiome to more bioavailable compounds, the urolithins, A and B, which are the main metabolites found in plasma and urine (Cerda et al, Eur. J. Nutr., 43 (2004), pp. 205-220).

[0008] Individuals display large variances (J. Agric. Food Chem., 62 (2014), pp. 6535-6538) in urolithin production capacity due to differences in the intestinal microbiome that is responsible for ellagitannins metabolism.

[0009] According to a new study by A. Cortes-Martin, et al., (Food Funct., 9 (2018), pp. 4100- 4106) only around 40 per cent of elderly individuals may hold the gut microbes to transform ellagitannins from pomegranate into urolithin A. Further, according to A. Cortes-Martin, et al, a person’s ability to produce Urolithin A invariably depends on presence of required gut microbiome and further varies with age, health status, and dietary intake.

[0010] Therefore, European Journal of Clinical Nutrition 76, 297-308 (2022) reported that differences in gut microbiome and diet can be overcome via direct dietary Urolithin A supplementation which boosts mitochondrial and muscle function and suggested that the direct supplementation with Urolithin A compound significantly increased plasma levels and provided a >6-fold exposure to Urolithin A vs. pomegranate juice, and thus helps in overcoming these individual limitations.

[0011] Backed by growing interest, several research groups started to study the role and relevance of direct supplementation with Urolithin A instead of Urolithin A precursors such as pomegranate and other fruit juices.

[0012] There is ample patented literature available on use of Urolithin A in a composition and methods of using these compositions as discussed herein below.

[0013] US 11903922 provides a method of treating a disease or a condition selected from the group consisting of stress, a stress-induced disorder, a stress-related disorder, and an anxiety disorder, comprising: administering to a subject in need thereof a therapeutically effective amount of a urolithin or a salt thereof, wherein the stress-induced or stress-related disorder is stress-induced or stress-related cognitive deficit, stress-induced or stress-related cognitive dysfunction, stress-induced or stress-related mood disorder, or stress-induced or stress-related anxiety. US10695320 and US11,925,616 discloses compositions comprising a medium chain triglyceride and a urolithin, and a method of improving physical endurance; inhibiting or retarding physical fatigue; or reducing muscle fatigue; comprising administering to a human in need thereof an effective amount of a composition comprising: a) medium-chain triglyceride; wherein the medium-chain triglyceride constitutes 30-80% w / w of the composition; b) micronized urolithin A, having a D50 size in a range of 0.5 to 50 pm, and a D90 size in a range of 5 to 100 pm; and c) lecithin, wherein the lecithin constitutes 1-40% w / w of the composition; wherein the composition is formulated for oral administration.

[0014] US 11878964 teaches compositions comprising a source of protein and a urolithin and also a method of improving physical performance, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin and a source of protein; wherein the physical performance is walking or running.

[0015] A method of treating a neurodegenerative disease, comprising: administering to a subject in need thereof a therapeutically effective amount of urolithin A, thereby treating the neurodegenerative disease; wherein the effective amount of urolithin A is about 140 mg to about 8,400 mg; and the neurodegenerative disease is selected from the group consisting of AIDS dementia complex, Alzheimer's disease, dementia with Lewy bodies, Huntington's disease, and Parkinson's disease is claimed in US 11234960.

[0016] A composition comprising: nicotinamide riboside, and urolithin A or a salt thereof; wherein nicotinamide riboside is present in an amount of 20 mg to 5 g; and urolithin A is present in an amount of 10 mg to 5 g is reported in US 11166972. US’972 further discloses that these compositions are useful as medicaments, for example, for treating muscle-related pathological conditions, neurodegenerative diseases, and / or mitochondrial diseases, and as dietary supplements, functional foods and beverages, and as specialized nutrition or medical foods.

[0017] US11166937 discloses compositions comprising compounds or precursors to compounds which may be used for a variety of therapeutic applications including, for example, treating and / or preventing a disease or disorder related to reduced or inadequate mitochondrial activity, including aging or stress, diabetes, obesity, and neurodegenerative diseases.

[0018] A method of increasing autophagy, comprising the step of orally administering to a healthy human in need thereof an effective amount of urolithin A, or a pharmaceutically acceptable salt thereof, wherein the healthy human is not suffering from cancer or predisposed to developing cancer, autophagy in the healthy human is increased, and the effective amount is about 70 mg to about 1050 mg, is reported in US11020373.

[0019] US10442784 reports a composition comprising: a) a source of protein; and b) urolithin A. US10988453 discloses and claims a composition comprising: a) a source of protein and b) micronized urolithin A; wherein the micronized urolithin A has a D50 size in the range 0.5 to 50 pm, and a D90 size in the range 5 to 100 pm, wherein the protein is selected from intact pea protein, intact pea protein isolates, intact pea protein concentrates, milk protein isolates, milk protein concentrates, casein protein isolates, casein protein concentrates, whey protein concentrates, whey protein isolates, sodium or calcium caseinates, whole cow's milk, partially or completely defatted milk, soy protein isolates and soy protein concentrates, and combinations thereof.

[0020] A method of improving a cognitive function, comprising: administering to a subject in need thereof an effective amount of a urolithin thereby improving the cognitive function is disclosed in US10857126.

[0021] As is evident from the above, Urol ethin A is reported to have wider therapeutic indications. Further, in 2018, the US Food and Drug Administration listed urolithin A as a safe ingredient for food or dietary supplement products having content in the range of 250 mg to one gram per serving.

[0022] Further, active investigation for the production of extracts containing a wide range of ellagitannins and phytochemicals is still ongoing research to arrive at an efficient extraction process, albeit there were multiple extraction processes reported, as discussed below.

[0023] EP1967079 Bl discloses a process of producing water soluble punicalagins rather than solvent soluble ellagic acid having the broad spectrum of the different natural organic chemicals which are present in the whole pomegranate fruit, e.g. both ellagitannins and anthocyanins.

[0024] IN238970 discloses extraction process of powdering the dried peel followed by organic solvent extraction, concentration and vacuum distillation. The extract thus obtained from the process has 78 % of phenolic content. WO2018134848 discloses preparation of aqueous, alcoholic and hydroalcoholic extracts of pomegranate peel, wherein, aqueous extracts are obtained with 9% Punicalagins & 4% Proanthocyanidins.

[0025] W02015103760 discloses pomegranate skin extract which comprises at least 30%, preferably 40%, more preferably 50%, still more preferably 60% of punicalagin, obtained by extracting pomegranate skin with a water-alcohol solution, such as an aqueous solution of ethanol.

[0026] US20060211635 discloses compositions of purified and biologically active ellagitannins provided by separation from pomegranate husk using a method of extraction and purification using a solid polymeric adsorbent and the uses of the said compounds.

[0027] US2022 / 0362139 claims an extract of Punica granatum comprising: i. at least 10% by weight of the total weight of the dry extract, of punicalagins; and ii. at least 10% by weight of the total weight of the dry extract, of ellagic acid-; and wherein the weight ratio of ellagic acid / punicalagins is ranging from 0.5 to 2.

[0028] US20230030835 discloses a method of enhanced production of urolithin B from pomegranate polyphenols by administering a composition comprising a synergistic mixture of pomegranate polyphenols, lycopene and quercetin.

[0029] US8658220 discloses processes for producing an extract containing phytochemicals from pomegranates which process comprise providing pomegranate solids, such as the pericarp, inner membrane and seeds; creating a mixture comprising the pomegranate solids in an aqueous solution; adding enzymes to the mixture in an amount sufficient to at least partially degrade the pomegranate solids; heating the mixture to a temperature that permits the maximum rate of catalysis of the enzyme; maintaining the temperature of the heated mixture for a time sufficient to allow at least partial degradation of the pomegranate solids; and removing residual insoluble solid materials from the mixture.

[0030] The foregoing literature have shown that the extracts of Punica granatum contains many phytochemicals, such as anthocyanins, ellagitannins, gallic acid, lignans along with other phytochemicals such as Gallotannins, Ferulic acid, Catechins, Quercetin, fatty acids, organic and phenolic antioxidants. Also, a cursory review of the above prior arts conclusively confirms that Urolithin A is the important active metabolite of the ellagitannins rich foods such as pomegranate juice. However, Urolithin A is produced by gut microflora and not everyone has a right microbiome capable of producing Urolithin A. Also, Urolithin A producers were distinguished by a significantly higher gut microbiome diversity and ratio of Firmicutes to Bacteroides. Therefore, ingestion of pomegranate juice or any other sources such as strawberries, walnut kernels, and peanuts having ellagitannins and ellagic acid by a person which has no or insufficient gut microbiome cannot produce Urolithin A and hence unable to achieve the desired therapeutic requirements. Moreover, it would be a safe to provide an extract as a composition containing Urolithin A within the extract itself rather than opting for a synthetic supplement.

[0031] In the light of the foregoing, it is an objective of the present invention to provide compositions of Punica granatum whole spectrum extract (PWSE) comprising Urolithin A.

[0032] It is another objective of the present invention to provide compositions comprising Punica granatum whole spectrum extract (PWSE), which is selectively enriched with Punicagalins, Ellagic acid and Urolethin A, to provide the desired Urolithin A within the natural extract itself thereby overcoming the need for external supplementation.

[0033] It is another objective of the invention to provide process for preparation of Punica granatum whole spectrum extract (PWSE) selectively enriched with Punicagalins, Ellagic acid and Urolethin A.

[0034] Definitions:

[0035] Probiotic blend (PB): Probiotic blend as used in the present invention is a proprietary blend which consists of Lactobacillus acidophilus, Bifidobacterium lactis, Bifidobacterium infantis, Lactobacillus plantarum, L. salivarius and B. longum.

[0036] Punica granatum whole spectrum extract (PWSE): Punica granatum whole spectrum extract (PWSE) as used herein in the entire specification comprises Punicagalins, Ellagic acid, Urolethin A, gallic acid, tannins and polyphenols.

[0037] Punica granatum whole spectrum extract selectively ‘enriched with’ or “enriched to comprise”: The phrase “Punica granatum whole spectrum extract selectively ‘enriched with’ or “enriched to comprise” as used herein in the entire specification means and includes that the extract is particularly enriched with or enriched to comprise Punicagalins, Ellagic acid and Urolethin A, in addition to the presence of gallic acid, tannins and polyphenols.

[0038] Summary of the invention:

[0039] In line with the above objectives, the present invention provides compositions of Punica granatum whole spectrum extract (PWSE) comprising Urolithin A, for improving general health, overall wellbeing and longevity.

[0040] According to an aspect, the invention provides compositions of Punica granatum whole spectrum extract (PWSE) selectively enriched with Punicagalins, Ellagic acid and Urolethin A, for improving general health, overall wellbeing and longevity. The compositions of the invention provide the desired Urolithin A within the natural extract itself thereby overcoming the need for external supplementation. The compositions of Punica granatum whole spectrum extract provided according to the present invention not only comprises Punicagalins, Ellagic acid, Urolethin A, but also comprise gallic acid, tannins and other polyphenols.

[0041] The compositions of the present invention enable the ‘Non-Urolithin A Producers’ to get the Urolithin A within the natural extracts of the pomegranate thereby positively reduces the need for the external supplementation of Urolithin A.

[0042] In an additional aspect, the invention provides process for preparation of composition of Punica granatum whole spectrum extract selectively enriched to comprise Urolethin A, Punicagalins and Ellagic acid by extracting with a source of probiotic.

[0043] In a further aspect, the present invention provides compositions comprising a) Punica granatum whole spectrum extract (PWSE) selectively enriched to comprise Punicagalins, Ellagic acid and Urolethin A; b) at least one herb / herbal extracts selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera. Bacopa monnieri. Asparagus racemosus. Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda and c) optionally fortified with vitamins and minerals.

[0044] In yet another aspect, the present invention provides compositions comprising Punica granatum whole spectrum extract (PWSE) selectively enriched to comprise Punicagalins, Ellagic acid and Urolethin A, fortified with one or more of vitamins such as Vitamin A, Vitamin B5, Vitamin B6, Vitamin B12 and other B Vitamins, Vitamin C, Vitamin D and Vitamin K and minerals such as Na, K, Mg, Ca and other essential minerals.

[0045] In a further aspect, the invention provides a method of inhibiting CD38 levels and increasing NAD levels in a human subject, which method comprises administering a therapeutically effective amounts of a composition comprising Punica granatum whole spectrum extract (PWSE) selectively enriched to comprise Punicagalins, Ellagic acid and Urolethin A, to the subject in need thereof.

[0046] In yet another aspect, the invention provides a method of increasing NAD levels in a human subject, which method comprises administering a therapeutically effective amounts of a composition comprising Punica granatum whole spectrum extract (PWSE) selectively enriched to comprise Punicagalins, Ellagic acid and Urolethin A, to the subject in need thereof.

[0047] Detailed description of the present invention:

[0048] The invention will now be described in detail in connection with certain preferred and optional embodiments so that various aspects thereof may be more fully understood and appreciated.

[0049] Source of biological material:

[0050] The biological material, Punica granatum used in the present invention are Bhagwa as well as Ganesh varieties procured from Nasik, India. The Bhagwa variety of pomegranate is a soft-seeded fruit with a bright red rind and arils and sweet, tangy flavor that is popular in India. The Ganesh variety of pomegranate is a medium-sized fruit with a yellowish red rind, pink flesh, and soft seeds. It is a popular variety in Maharashtra and is the commercial cultivar of the state.

[0051] The present invention provides compositions of Punica granatum whole spectrum extract (PWSE) which comprises Urolithin A.

[0052] According to an embodiment, the invention provides compositions of Punica granatum whole spectrum extract (PWSE) selectively enriched with Punicagalins, Ellagic acid and Urolethin A, to provide the desired Urolithin A within the natural extract itself thereby overcoming the need for external supplementation. The compositions of the present invention useful for improving general health, overall wellbeing and longevity, by preventing age related disorders. NAD+ levels naturally decline with age, and this decline is linked to numerous age-related disorders and a decrease in lifespan. Therefore, boosting NAD+ levels in a subject is essential to prevent age related disorders and improves general health, overall wellbeing and longevity.

[0053] Thus, the compositions of the present invention prevents age related disorders, by increasing NAD+levels, such as impaired cognitive functions including neurodegenerative disorders with symptoms of memory loss, cognitive decline, motor dysfunction, neuroinflammation, musculoskeletal degeneration, mitochondrial diseases with symptoms of muscle fatigue, exercise intolerance, organ dysfunction and metabolic disorders with symptoms of insulin resistance, abnormal fat accumulation, fatigue as well as other age related common symptoms such as mood disturbances, muscle weakness, poor sleep, and slower recovery from illness, which are the hallmark signs of NAD+depletion and further improves general health and longevity.

[0054] In one of the preferred embodiments, the invention provides compositions of Punica granatum whole spectrum extract (PWSE) extract comprising Urolithin A in the range of 0.2% - 10%.

[0055] In one of the preferred embodiments, the invention provides compositions of Punica granatum whole spectrum extract (PWSE) extract selectively enriched to comprise Punicagalins in the range of 2% - 35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, which is naturally produced within the extract, as exemplified herein below through the examples 3 to 8.

[0056] Punica granatum whole spectrum extract (PWSE) composition provided according to the present invention comprises good amounts of polyphenols such as gallic acid; Tannins and other Polyphenols, in addition to the enriched amounts of Punicagalins, Ellagic acid and Urolethin A.

[0057] Accordingly, in another preferred embodiment, the invention provides compositions comprising Punica granatum whole spectrum extract (PWSE) which comprises Punicagalins in the range of 2.0% - 35.0%; Ellagic acid in the range of 0.5% - 14.0%; Urolethin A in the range of 0.2% - 10.0%; Galic acid in the range of 0.1% - 2.5%; Tannins in the range of 5.0% - 50.0%; and other Polyphenols in the range of 1.0% - 15.0%. In a further embodiment, the present invention provides a composition comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%; and optionally with at least one ingredient selected from the group consisting of a herb / herbal extract, vitamins, minerals, pharmaceutically or nutraceutically or dietically acceptable excipients, carriers, and diluents, for improving general health, overall wellbeing and longevity.

[0058] The ‘herb / herbal extract’, as referred to in the compositions of the invention are listed under “Rasayana” in Ayurveda, which is selected from the group consisting of Acorus calamus, Allium sativum, Aloe vera, Argyreia speciosa, Asparagus racemosus, Azadirachta indica, Bacopa monerri, Boerhavia diffusa, Cissampelos pareira, Commiphora mukul, Convolvulus phuricaulis Curculigo orchlodes, Curcuma longa, Despmodium gangeticum, Dioscora bulbifera, Emblica officinalis, Emblica ribes, Glycyrrhiza glabra, Gmelina arborea, Hemidesmus indicus, lopmoea digitata, Leptidenia reticulate, Piper longum, Plumbego zeylenica, Psorelea corylifolia, Perocarpus marsupium, Semecarpus anacardium, Sida spinosa, Solanum nigrum, Terminalia bellerica, Terminalia chebula, Tinospora cordifolia, Withania somnifera.

[0059] In yet another embodiment, the present invention provides compositions comprising a) Punica granatum whole spectrum extract (PWSE) selectively enriched to comprise Punicagalins, Ellagic acid and Urolethin A; b) at least one herb / herbal extracts selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri. Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda and c) optionally fortified with one or more of vitamins and minerals.

[0060] The vitamins may be selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin B12 and other B Vitamins, Vitamin C, Vitamin D and Vitamin K and the minerals are selected from the group consisting of Na, K, Mg, Ca and other essential minerals. In a preferred embodiment, the invention provides process for preparation of Punica granatum whole spectrum extract (PWSE), which process comprises; a step of extracting the Punica granatum whole fruit with a source of probiotic.

[0061] The Punica granatum whole fruit may be fresh fruit or dried fruit which includes fruit rind, seeds containing arils, albedo, membrane and other fruit parts of Punica granatum.

[0062] In a further embodiment, the source of probiotic is selected from the group consisting of milk, yogurt and a probiotic blend.

[0063] In one of the embodiments, the probiotic blend is a proprietary blend that consisting of Lactobacillus acidophilus, Bifidobacterium laclis. Bifidobacterium infanlis. Lactobacillus plantarum, L. salivarius and B. longum that can be used in the process in appropriate amounts to obtain Urolithin A in the range of 0.2% - 10.0% as exemplified in the specification.

[0064] Accordingly, in one of the preferred embodiments, the invention provides process for preparation of Punica granatum whole spectrum extract (PWSE) selectively enriched with Punicagalins, Ellagic acid and Urolethin A, which comprises the steps of; a) extracting the Punica granatum whole fruit with a solution containing mixture of milk and water for 6-10 h with intermittent stirring at ambient temperature; b) filtering the liquid extract from the remaining fruit mass and evaporating the liquid extract to isolate the Punica granatum milk extract powder (PME); c) subj ecting the remaining fruit mass to repeated aqueous extractions at a temperature of 90-95°C; d) filtering the aqueous extract from the remaining fruit mass; e) subj ecting the remaining fruit mass to repeated alcohol extractions at a temperature of 60-65°C; f) filtering the alcohol extract from the remaining fruit mass, combining the aqueous and alcohol extracts and isolating the Punica granatum extract powder enriched with high polarity phytoconstituents-PEHPP by evaporating the combined extract; and g) combining the Punica granatum milk extract powder of step b) and Punica granatum extract powder enriched with high polarity phytoconstituents-PEHPP of step f) to obtain Punica granatum whole spectrum extract (PWSE). According to the above embodiment, the invention provides compositions comprising Punica granatum whole spectrum extract (PWSE) extract which may encompass the source of probiotic i.e., milk solids. However, in the light of prevalence of lactose intolerance in many consumers, the present invention provides another alternate process for preparation of Punica granatum whole spectrum extract (PWSE), extracted with a proprietary probiotic blend, herein after referred as PWSpE.

[0065] Accordingly, in another embodiment, the invention provides process for preparation of Punica granatum whole spectrum extract (PWSpE) selectively enriched with Punicagalins, Ellagic acid and Urolethin A, extracted with a proprietary probiotic blend which process comprises the steps of; a) extracting the dried Punica granatum powder with a solution containing mixture of probiotic blend and water for 16-20 h with intermittent stirring at ambient temperature; b) filtering the liquid extract from the remaining fruit mass and isolating the Punica granatum probiotic extract powder (PPE) by evaporating the liquid extract; c) subj ecting the remaining fruit mass to repeated aqueous extractions at a temperature of 90-95°C; d) filtering the aqueous extract from the remaining fruit mass; e) subj ecting the remaining fruit mass to repeated alcohol extractions at a temperature of 60-65°C; f) filtering the alcohol extract from the remaining fruit mass, combining the aqueous and alcohol extracts to isolate the Punica granatum extract powder post probiotic treatment- PEPPT by evaporation of the combined extract; and g) combining the Punica granatum probiotic extract powder (PPE) of step b) and Punica granatum extract powder post probiotic treatment - PEPPT of step f) to obtain Punica granatum whole spectrum extract (PWSpE).

[0066] The probiotic blend as used in the process is a proprietary blend that consisting of Lactobacillus acidophilus, Bifidobacterium laclis. Bifidobacterium infanlis. Lactobacillus plantarum, L. salivarius and B. longum. Thus, the process of the present invention achieves Punica granatum whole spectrum extract (PWSE) selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%.

[0067] The present invention also demonstrated under examples 1 and 2, that the conventional extraction methods of Punica granatum (hydroalcoholic extracts) do not yield an extract or a composition that contain Urolithin A, however, yield a composition that contains punicalagins, ellagic acid, gallic acid, tannins and other polyphenols.

[0068] Therefore, the composition of Punica granatum whole spectrum extract provided with Urolithin A, according to the present invention is hitherto unreported in the literature. The composition of Punica granatum whole spectrum extract containing Urolithin A or a composition of Punica granatum whole spectrum extract containing Urolithin A along with Punicagalins and Ellagic acid is presumed to be capable of further conversion of Punicagalins and Ellagic acid synergistically into Urolithins due to the traces of source of probiotic present in the composition and thus helps Non-producers of Urolithin A to successfully utilize benefits of the whole spectrum extract.

[0069] In one of the embodiments, the invention successfully demonstrated that the extracts prepared in accordance with the present invention boosts NAD levels & and inhibits CD38 levels, in comparison to conventional extracts (tables 1 and 2).

[0070] According to this embodiment, activation of Nicotinamide Adenine Dinucleotide (NAD+) for various extracts of Punica granatum containing punicalagins, ellagic acid, and urolithin A was evaluated by In vitro assay in C2C12 mouse skeletal myoblasts (ATCC Cat# CRL- 1772) using NMN as positive control. NAD measurement was performed using the Promega NAD / NADH-Glo™ kit (Cat# G9071).

[0071] The percent increase of NAD over vehicle control as a result of sample treatment was calculated by the following formula:

[0072] % Increase in NAD over vehicle control = [(NAD concn, in sample-NAD concn, in vehicle control)] / NAD concn, in vehicle control x 100.

[0073] The results obtained are presented in table 1. The percent increase of NAD over control clearly indicate that NAD activation is increased in all the extracts of Punica granatum containing punicalagins, ellagic acid, and urolithin A over control, as per the examples of 3 to 8 in comparison to example 1 and 2, wherein the extract does not contain Urolithin A. These results clearly indicate synergistic effect of Punica granatum extract containing punicalagins and ellagic acid with Urolithin A.

[0074] Similarly, CD38 inhibition activity for various extracts of Punica granatum containing punicalagins, ellagic acid, and urolithin A was also evaluated in L6 Rat skeletal myoblasts (ATCC Cat# CRL-1458). The % Inhibition of CD38 activity was calculated by the following formula:

[0075] %Inhibition of CD38 activity = [(Normalized RFU of Enzyme alone) - (Normalized RFU of Sample)] / Normalized RFU of Enzyme alone x 100

[0076] The results are presented in table 2. The percent increase of inhibition of CD38 clearly indicate that CD38 inhibition activity is increased in all the extracts of Punica granatum containing punicalagins, ellagic acid, and urolithin A as per the examples of 3 to 8 in comparison to example 1 and 2, wherein the extract does not contain Urolithin A. These results clearly indicate synergistic effect of Punica granatum extract containing punicalagins and ellagic acid with Urolithin A.

[0077] Therefore, the Punica granatum whole spectrum extract containing punicalagins and ellagic acid with Urolithin A exhibits anti-inflammatory effect, Testosterone boosting effect, endurance and muscle building effects Pre- and Post workout by significantly increasing NAD levels and by inhibiting CD38.

[0078] Accordingly, in yet another embodiment, the invention provides a method of increasing NAD levels in a human subject, which method comprises supplementing the human subject in need thereof, a therapeutically effective amounts of a composition comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%; and optionally with at least one ingredient selected from the group consisting of a herb / herbal extract, vitamins, minerals, pharmaceutically or nutraceutically or dietically acceptable excipients, carriers, and diluents.

[0079] In certain embodiments, the herb or herbal extract is selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera. Bacopa monnieri. Asparagus racemosus. Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda and optionally fortified with vitamins and minerals.

[0080] The herb / herbal extract, as referred to in the ‘methods of the invention’ are listed under “Rasayana” in Ayurveda, which is selected from the group consisting of Acorus calamus, Allium sativum, Aloe vera, Argyreia speciosa, Asparagus racemosus, Azadirachta indica, Bacopa monerri, Boerhavia diffusa, Cissampelos pareira, Commiphora mukul, Convolvulus phuricaulis Curculigo orchlodes, Curcuma longa, Despmodium gangeticum, Dioscora bulbifera, Emblica officinalis, Emblica ribes, Glycyrrhiza glabra, Gmelina arborea, Hemidesmus indicus, lopmoea digitata, Leptidenia reticulate, Piper longum, Plumbego zeylenica, Psorelea corylifolia, Perocarpus marsupium, Semecarpus anacardium, Sida spinosa, Solanum nigrum, Terminalia bellerica, Terminalia chebula, Tinospora cordifolia, Withania somnifera.

[0081] According to yet another embodiment, a method of inhibiting CD38 levels in a human subject, which method comprises supplementing the human subject in need thereof, a therapeutically effective amounts of a composition comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, and optionally with at least one ingredient selected from the group consisting of a herb / herbal extract, vitamins, minerals, pharmaceutically or nutraceutically or dietically acceptable excipients, carriers, and diluents.

[0082] In certain embodiments, the herb or herbal extract is selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri, Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda and optionally fortified with vitamins and minerals.

[0083] Thus according to one of the additional embodiments, the invention provides compositions comprising a) Punica granatum whole spectrum extract (PWSE) selectively enriched with Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, may be formulated into a dosage form along with b) at least one herb / herbal extract selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri, Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda and c) optionally fortified with vitamins and minerals.

[0084] The ‘herb / herbal extract’, as referred to “in the compositions” of the invention are listed under “Rasayana” in Ayurveda, which is selected from the group consisting of Acorus calamus, Allium sativum, Aloe vera, Argyreia speciosa, Asparagus racemosus, Azadirachta indica, Bacopa monerri, Boerhavia diffusa, Cissampelos pareira, Commiphora mukul, Convolvulus phuricaulis Curculigo orchlodes, Curcuma longa, Despmodium gangeticum, Dioscora bulbifera, Emblica officinalis, Emblica ribes, Glycyrrhiza glabra, Gmelina arborea, Hemidesmus indicus, lopmoea digitata, Leptidenia reticulate, Piper longum, Plumbego zeylenica, Psorelea corylifolia, Perocarpus marsupium, Semecarpus anacardium, Sida spinosa, Solanum nigrum, Terminalia bellerica, Terminalia chebula, Tinospora cordifolia, Withania somnifera.

[0085] The vitamins may be selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin B 12 and other B Vitamins, Vitamin C, Vitamin D and Vitamin K and minerals may be selected from the group consisting of Na, K, Mg, Ca and other essential minerals.

[0086] In one another embodiment of the invention, the invention provides compositions of Punica granatum whole spectrum extract (PWSE) which comprises Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, fortified with one or more vitamins selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin B12, and other B Vitamins, Vitamin C, Vitamin D and Vitamin K and one or more minerals selected from the group consisting of Na, K, Mg, Ca and other essential minerals. The other B vitamins include thiamine (Bl), riboflavin (B2), niacin (B3), biotin (B7), and folate (B9).

[0087] In another embodiment of the invention, the composition of Punica granatum whole spectrum extract (PWSE) comprising punicalagins and ellagic acid with Urolithin A, may be formulated into a dosage form selected from powder, a tablet, a capsule, a soft chewable tablet or liquid form, beverage, food product, dietary supplement, or any other suitable forms using suitable conventional pharmaceutical / nutraceutical excipients, as exemplified under example 11. In yet another embodiment, the invention provides use of compositions comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, for increasing NAD levels in a human subject.

[0088] In a further embodiment, the invention provides use of compositions comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, b) at least one herb / herbal extract selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri. Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda and c) optionally fortified with vitamins and minerals, for increasing NAD levels in a human subject.

[0089] In yet another embodiment, the invention provides use of compositions comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, for inhibiting CD38 levels in a human subject.

[0090] In a further embodiment, the invention provides use of compositions comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, b) at least one herb / herbal extract selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri, Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda and c) optionally fortified with vitamins and minerals, for inhibiting CD38 levels in a human subject.

[0091] The ‘herb / herbal extract’, as referred to in the ‘use of the compositions’ are listed under “Rasayana” in Ayurveda, which is selected from the group consisting of Acorus calamus, Allium sativum, Aloe vera, Argyreia speciosa, Asparagus racemosus, Azadirachta indica, Bacopa monerri, Boerhavia diffusa, Cissampelos pareira, Commiphora mukul, Convolvulus phuricaulis Curculigo orchlodes, Curcuma longa, Despmodium gangeticum, Dioscora bulbifera, Emblica officinalis, Emblica ribes, Glycyrrhiza glabra, Gmelina arborea, Hemidesmus indicus, lopmoea digitata, Leptidenia reticulate, Piper longum, Plumbego zeylenica, Psorelea corylifolia, Perocarpus marsupium, Semecarpus anacardium, Sida spinosa, Solanum nigrum, Terminalia bellerica, Terminalia chebula, Tinospora cordifolia, Withania somnifera.

[0092] In a further embodiment, the invention provides use of compositions comprising Punica granatum whole spectrum extract selectively enriched with Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, fortified with one or more vitamins selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin B12 and other B Vitamins, Vitamin C, Vitamin D and Vitamin K and one or more minerals selected from the group consisting of Na, K, Mg, Ca and other essential minerals, for increasing NAD levels or for inhibiting CD38 levels in a human subject.

[0093] Those of ordinary skilled in the art will appreciate that changes could be made to the embodiments described above without departing from the broad inventive concept thereof. It is understood, therefore, that this invention is not limited to the particular embodiments or examples disclosed herein but is intended to cover modifications within the objectives and scope of the present invention as defined in the specification. The examples are given solely for illustration and are not to be construed as limitations of the present disclosure, as many variations thereof possible without departing from the spirit of the disclosure.

[0094] Examples:

[0095] Example 1:

[0096] Preparation of Punica granatum 70% aqueous ethanol extract from dried mixture of fruit rind and seed including arils (POM-1)

[0097] Dried Punica granatum powder consisting of dried fruit rind and seeds containing arils (30 Kg) was added to 70% aq. ethanol (150 Litre) and stirred at 60-65°C for 3 h. The mixture was filtered through celite, and the extraction process was repeated with 70% aq. ethanol (2 x 150 Litre) at 60-65°C for 3 h. To the combined 70% aqueous ethanol extract was evaporated under reduced pressure and dried using spray dryer to give the product as pale brown to reddish brown color solid (8.0 Kg). The product was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urolithin A and other phytochemicals. As per the HPLC analysis, it was found that Punica granatum 70% aqueous ethanol extract obtained from dried mixture of fruit rind and seeds containing arils contains 18.16% of punicalagins, 3.08% of ellagic acid, and 0.61% of gallic acid. Tannin content of Punica granatum 70% aqueous ethanol extract obtained by example 1 is 37.15% w / w and other polyphenol content was found to be 16.36%, while Urolethin A was found to be absent.

[0098] Example 2:

[0099] Preparation of Punica granatum 90% aqueous ethanol extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts (POM-2)

[0100] Fresh Punica granatum fruit consisting of fruit rind, seeds containing arils, albedo, membrane and other fruit parts were cut in small pieces (30 Kg) and then added to 90% aq. ethanol (150 Litre) and stirred at 90-95°C for 3 h. The mixture was filtered through celite, and the extraction process was repeated with 90% aq. ethanol (2 x 150 Litre) at 90-95°C for 3 h. To the combined 90% aqueous ethanol extract was evaporated under reduced pressure and dried using spray dryer to give the product as brown to reddish brown color solid (1.9 Kg). The product was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urolethin A and other phytochemicals. As per the HPLC analysis, it was found that Punica granatum 90% aqueous ethanol extract obtained from Fresh Punica granatum fruit consisting of fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 14.30% of punicalagins, 2.98% of ellagic acid, and 0.58% of gallic acid. Tannin content of Punica granatum 90% aqueous ethanol extract obtained by example 2 is 16.32% w / w and other polyphenol content was found to be 3.66%, Urolethin A was found to be absent.

[0101] Example 3:

[0102] Preparation of Punica granatum whole spectrum extract from dried mixture of fruit rind and seeds including arils

[0103] Part I : Dried Punica granatum powder consisting of dried fruit rind and seeds containing arils (20 Kg) was added to 20Kg of solution containing milk and water in ratio of 1 :2 and kept for 6-8 h with intermittent stirring at ambient temperature. The liquid extract was filtered through celite, and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to brown colored Punica granatum milk extract powder - PME (1.8 Kg). As per the HPLC analysis, it was found that Punica granatum milk extract obtained from dried mixture of fruit rind and seeds containing arils contains 9.86% of punicalagins, 2.78% of ellagic acid, and 0.27% of gallic acid. Tannin content of Punica granatum milk extract obtained by example 3 part I is 10.05% w / w. Other polyphenol content was found to be 3.37% and Urolethin A was found to be 2.97%.

[0104] Part II: The remaining fruit mass was then extracted with water (100 Litre) at 90-95°C for 3 h. Post completion of water extraction, the liquid extract was filtered through celite, and the remaining fruit mass was again extracted with water (100 Litre) at 90-95°C for 3 h for second time. Post completion of water extraction, the liquid water extract was filtered through celite, and combined with first water extract derived in the process. Subsequently, the remaining fruit mass was extracted with alcohol (2 X 100 Litre) at 60-65°C for 3 h. Both alcohol extract derived were then combined with combined water extracts derived earlier and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to reddish brown colored Punica granatum enriched extract powder with high polarity phytoconstituents - PEHPP (5.5 Kg). As per the HPLC analysis, it was found that Punica granatum enriched extract obtained from dried mixture of fruit rind and seeds (PEHPP) containing arils contains 15.84% of punicalagins, 5.18% of ellagic acid, and 0.79% of gallic acid. Tannin content of Punica granatum enriched extract (PEHPP) obtained by example 3 part II is 27.54% w / w. Other polyphenol content was found to be 7.55% and Urolethin A was found to be 0.17%.

[0105] Part III: Punica granatum milk extract powder (PME) and Punica granatum enriched extract powder with high polarity phytoconstituents (PEHPP) were mixed in ratio of 1 :3 and named as Punica granatum whole spectrum extract (PWSE). The product - PWSE was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urolethin A and other phytochemicals. As per the HPLC analysis, it was found that PWSE contains 14.68% of punicalagins, 6.09% of ellagic acid, and 0.73% of gallic acid. Tannin content of Punica granatum enriched extract (PWSE) obtained by example 3 part III is 24.01% w / w. Other polyphenol content was found to be 6.17% and Urolethin A was found to be 0.84%. Example 4:

[0106] Preparation of Punica granatum whole spectrum extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts

[0107] Part I : Fresh Punica granatum fruit consisting of fruit rind, seeds containing arils, albedo, membrane and other fruit parts were cut in small pieces (20 Kg) and added to 20Kg of solution containing milk and water in ratio of 1 :2 and kept for 6-8 h with intermittent stirring at ambient temperature. The liquid extract was filtered through celite, and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to brown colored Punica granatum milk extract powder - PME (2.6 Kg). As per the HPLC analysis, it was found that Punica granatum milk extract obtained from fresh fruits of fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 11.92% of punicalagins, 4.01% of ellagic acid, and 1.00% of gallic acid. Tannin content of Punica granatum milk extract obtained by example 4 part I is 16.10% w / w. Other polyphenol content was found to be 2.87% and Urolethin A was found to be 2.18%.

[0108] Part II: The remaining fruit mass was then extracted with water (100 Litre) at 90-95°C for 3 h. Post completion of water extraction, the liquid extract was filtered through celite, and the remaining fruit mass was again extracted with water (100 Litre) at 90-95°C for 3 h for second time. Post completion of water extraction, the liquid water extract was filtered through celite, and combined with first water extract derived in the process. Subsequently, the remaining fruit mass was extracted with alcohol (2 X 100 Litre) at 60-65°C for 3 h. Both alcohol extract derived were then combined with combined water extracts derived earlier and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to reddish brown colored Punica granatum enriched extract powder with high polarity phytoconstituents - PEHPP (1.8 Kg). As per the HPLC analysis, it was found that Punica granatum enriched extract powder obtained from fresh fruit including fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 18.04% of punicalagins, 7.23% of ellagic acid, and 0.94% of gallic acid. Tannin content of Punica granatum enriched extract obtained by example 4 part II is 27.05% w / w. Other polyphenol content was found to be 9.87% and Urolethin A was found to be 0.59%. Part III: Punica granatum milk extract powder (PME) and Punica granatum enriched extract powder with high polarity phytoconstituents (PEHPP) were mixed in ratio of 2: 1 and named as Punica granatum whole spectrum extract (PWSE). The product - PWSE was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urol ethin A and other phytochemicals. As per the HPLC analysis, it was found that PWSE contains 15.03% of punicalagins, 5.48% of ellagic acid, and 0.63% of gallic acid. Tannin content of Punica granatum enriched extract (PWSE) obtained by example 4 part III is 21.32% w / w. Other polyphenol content was found to be 5.66% and Urolethin A was found to be 1.07%.

[0109] Example 5:

[0110] Preparation of Punica granatum whole spectrum extract from dried mixture of fruit rind and seeds including arils seeded with Probiotic blend

[0111] Part I : Dried Punica granatum powder consisting of dried fruit rind and seeds containing arils in ratio of 4: 1 (20 Kg) was added to 20Kg of solution containing water and probiotic blend (PB) consisting of Lactobacillus acidophilus, Bifidobacterium laclis. Bifidobacterium infanlis. Lactobacillus plantarum, L. salivarius and A longum and kept for 16-18 h with intermittent stirring at ambient temperature. The liquid extract was filtered through celite, and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to brown colored Punica granatum probiotic extract powder - PPE (1.24 Kg). As per the HPLC analysis, it was found that Punica granatum probiotic extract (PPE) obtained from dried mixture of fruit rind and seeds containing arils contains 11.71% of punicalagins, 3.07% of ellagic acid, and 0.50% of gallic acid. Tannin content of Punica granatum probiotic extract obtained by example 5 part I is 14.05% w / w. Other polyphenol content was found to be 5.07% and Urolethin A was found to be 3.90%.

[0112] Part II: The remaining fruit mass was then extracted with water (100 Litre) at 90-95°C for 3 h. Post completion of water extraction, the liquid extract was filtered through celite, and the remaining fruit mass was again extracted with water (100 Litre) at 90-95°C for 3 h for second time. Post completion of water extraction, the liquid water extract was filtered through celite, and combined with first water extract derived in the process. Subsequently, the remaining fruit mass was extracted with alcohol (2 X 100 Litre) at 60-65°C for 3 h. Both alcohol extract derived were then combined with combined water extracts derived earlier and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to reddish brown colored Punica granatum enriched extract post probiotic treatment - PEPPT (5.10 Kg). As per the HPLC analysis, it was found that Punica granatum enriched extract obtained from dried mixture of fruit rind and seeds (PEHPP) containing arils contains 16.84% of punicalagins, 7.23% of ellagic acid, and 0.87% of gallic acid. Tannin content of Punica granatum enriched extract (PEPPT) obtained by example 5 part II is 29.04% w / w. Othar polyphenol content was found to be 7.95% and Urolethin A was found to be 0.84%.

[0113] Part III: Punica granatum probiotic extract powder (PPE) and Punica granatum enriched extract post probiotic treatment - PEPPT were mixed and named as Punica granatum whole spectrum probiotic extract (PWSpE). The product - PWSpE was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urolethin A and other phytochemicals. As per the HPLC analysis, it was found that PWSE contains 16.01% of punicalagins, 7.09% of ellagic acid, and 0.78% of gallic acid. Tannin content of Punica granatum whole spectrum probiotic extract (PWSpE) obtained by example 5 part III is 26.01% w / w. Other polyphenol content was found to be 7.17% and Urolethin A was found to be 1.39%.

[0114] Example 6:

[0115] Preparation of Punica granatum whole spectrum extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts

[0116] Part I : Fresh Punica granatum fruit consisting of consisting of fruit rind, seeds containing arils, albedo, membrane and other fruit parts were cut in small pieces (20 Kg) and was added to 20Kg of solution containing water and probiotic blend (PB) consisting of Lactobacillus acidophilus, Bifidobacterium lactis, Bifidobacterium infantis, Lactobacillus plantarum, L. salivarius, B. longum, L. in ratio of 1 :0.3 and kept for 16-18 h with intermittent stirring at ambient temperature. The liquid extract was filtered through celite, and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to brown colored Punica granatum probiotic extract powder - PPE (1.85 Kg). As per the HPLC analysis, it was found that Punica granatum probiotic extract obtained from fresh fruits of fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 14.12% of punicalagins, 4.53% of ellagic acid, and 1.34% of gallic acid. Tannin content of Punica granatum probiotic extract obtained by example 6 part I is 19.56% w / w. Other polyphenol content was found to be 2.87% and Urolethin A was found to be 2.18%.

[0117] Part II: The remaining fruit mass was then extracted with water (100 Litre) at 90-95°C for 3 h. Post completion of water extraction, the liquid extract was filtered through celite, and the remaining fruit mass was again extracted with water (100 Litre) at 90-95°C for 3 h for second time. Post completion of water extraction, the liquid water extract was filtered through celite, and combined with first water extract derived in the process. Subsequently, the remaining fruit mass was extracted with alcohol (2 X 100 Litre) at 60-65°C for 3 h. Both alcohol extract derived were then combined with combined water extracts derived earlier and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to reddish brown colored Punica granatum enriched extract post probiotic treatment - PEPPT (1.50 Kg). As perthe HPLC analysis, it was found that Punica granatum enriched extract powder obtained from fresh fruit including fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 19.20% of punicalagins, 8.64% of ellagic acid, and 0.99% of gallic acid. Tannin content of Punica granatum enriched extract obtained by example 6 part II is 30.27% w / w. Other polyphenol content was found to be 9.87% and Urolethin A was found to be 0.59%.

[0118] Part III: Punica granatum probiotic extract powder (PPE) and Punica granatum enriched extract post probiotic treatment - PEPPT were mixed and named as Punica granatum whole spectrum probiotic extract (PWSpE). The product - PWSpE was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urolethin A and other phytochemicals. As per the HPLC analysis, it was found that PWSpE contains 17.33% of punicalagins, 6.48% of ellagic acid, and 1.14% of gallic acid. Tannin content of Punica granatum whole spectrum probiotic extract (PWSpE) obtained by example 6 part III is 24.20% w / w. Other polyphenol content was found to be 5.66% and Urolethin A was found to be 1.65%.

[0119] Example 7:

[0120] Preparation of Punica granatum whole spectrum extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts Part I: Fresh Punica granatum fruit consisting of fruit rind, seeds containing arils, albedo, membrane and other fruit parts were cut in small pieces (20 Kg) and added to 20Kg of solution containing milk and water in ratio of 1 :4 and kept for 6-8 h with intermittent stirring at ambient temperature. The liquid extract was filtered through celite, and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to brown colored Punica granatum milk extract powder - PME (2.9 Kg). As per the HPLC analysis, it was found that Punica granatum milk extract obtained from fresh fruits of fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 16.18% of punicalagins, 7.15% of ellagic acid, and 1.40% of gallic acid. Tannin content of Punica granatum milk extract obtained by example 7 part I is 23.55% w / w. Polyphenol content was found to be 3.70% and Urol ethin A was found to be 4.01%.

[0121] Part II: The remaining fruit mass was then extracted with water (100 Litre) at 90-95°C for 3 h. Post completion of water extraction, the liquid extract was filtered through celite, and the remaining fruit mass was again extracted with water (100 Litre) at 90-95°C for 3 h for second time. Post completion of water extraction, the liquid water extract was filtered through celite, and combined with first water extract derived in the process. Subsequently, the remaining fruit mass was extracted with alcohol (2 X 100 Litre) at 60-65°C for 3 h. Both alcohol extract derived were then combined with combined water extracts derived earlier and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to reddish brown colored Punica granatum enriched extract powder with high polarity phytoconstituents - PEHPP (2.4 Kg). As per the HPLC analysis, it was found that Punica granatum enriched extract powder obtained from fresh fruit including fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 26.43% of punicalagins, 9.10% of ellagic acid, and 1.67% of gallic acid. Tannin content of Punica granatum enriched extract obtained by example 7 part II is 30.11% w / w. Polyphenol content was found to be 11.90% and Urolethin A was found to be 1.95%.

[0122] Part III: Punica granatum milk extract powder (PME) and Punica granatum enriched extract powder with high polarity phytoconstituents (PEHPP) were mixed in ratio of 1 : 1 and named as Punica granatum whole spectrum extract (PW SE). The product - PW SE was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urolethin A and other phytochemicals. As per the HPLC analysis, it was found that PWSE contains 20.90% of punicalagins, 8.07% of ellagic acid, and 1.57% of gallic acid. Tannin content of Punica granatum enriched extract (PWSE) obtained by example 7 part III is 26.52% w / w. Polyphenol content was found to be 7.61% and Urol ethin A was found to be 3.08%.

[0123] Example 8:

[0124] Preparation of Punica granatum whole spectrum extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts

[0125] Part I: Fresh Punica granatum fruit consisting of consisting of fruit rind, seeds containing arils, albedo, membrane and other fruit parts were cut in small pieces (20 Kg) and was added to 20Kg of solution containing water and probiotic blend (PB) consisting of Lactobacillus acidophilus, Bifidobacterium laclis. Lactobacillus plantarum and Lactobacillus fermentum in ratio of 1 :0.2 and kept for 16-18 h with intermittent stirring at ambient temperature. The liquid extract was filtered through celite, and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to brown colored Punica granatum probiotic extract powder - PPE (2.15 Kg). As per the HPLC analysis, it was found that Punica granatum probiotic extract obtained from fresh fruits of fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 19.02% of punicalagins, 6.03% of ellagic acid, and 1.48% of gallic acid. Tannin content of Punica granatum probiotic extract obtained by example 8 part I is 30.16% w / w. Polyphenol content was found to be 5.12% and Urol ethin A was found to be 5.88%.

[0126] Part IE The remaining fruit mass was then extracted with water (100 Litre) at 90-95°C for 3 h. Post completion of water extraction, the liquid extract was filtered through celite, and the remaining fruit mass was again extracted with water (100 Litre) at 90-95°C for 3 h for second time. Post completion of water extraction, the liquid water extract was filtered through celite, and combined with first water extract derived in the process. Subsequently, the remaining fruit mass was extracted with alcohol (2 X 100 Litre) at 60-65°C for 3 h. Both alcohol extract derived were then combined with combined water extracts derived earlier and then evaporated under reduced pressure and dried using spray dryer to give the product as light brown to reddish brown colored Punica granatum enriched extract post probiotic treatment - PEPPT (3.50 Kg). As per the HPLC analysis, it was found that Punica granatum enriched extract powder obtained from fresh fruit including fruit rind, seeds containing arils, albedo, membrane and other fruit parts contains 22.70% of punicalagins, 8.04% of ellagic acid, and 1.25% of gallic acid. Tannin content of Punica granatum enriched extract obtained by example 8 part II is 32.64% w / w. Polyphenol content was found to be 7.97% and Urolethin A was found to be 1.39%.

[0127] Part III: Punica granatum probiotic extract powder (PPE) and Punica granatum enriched extract post probiotic treatment - PEPPT were mixed and named as Punica granatum whole spectrum probiotic extract (PWSpE). The product - PWSpE was analyzed for Tannins, Punicalagins, Ellagic acid, Gallic acid, Urolethin A and other phytochemicals. As per the HPLC analysis, it was found that PWSpE contains 22.71% of punicalagins, 7.76% of ellagic acid, and 1.43% of gallic acid. Tannin content of Punica granatum whole spectrum probiotic extract (PWSpE) obtained by example 8 part III is 33.97% w / w. Polyphenol content was found to be 7.34% and Urolethin A was found to be 3.30%.

[0128] Example 9:

[0129] In vitro assay to evaluate the activation of Nicotinamide Adenine Dinucleotide (NAD+) for various extracts of Punica granatum containing punicalagins, ellagic acid, and urolithin A

[0130] Cell culture and Treatment: In a 48-well cell culture plate, an equal number of C2C12 mouse skeletal myoblasts (ATCC Cat# CRL-1772) were seeded. At 80-95% confluence, cells were induced to differentiate in DMEM medium (Himedia; Mumbai, India) supplemented with 1.9% Horse Serum (ATCC Cat# 30-2040) for an additional 7 days to form myotubes. After differentiation, the C2C12 myotubes were treated with or without different concentrations of test samples or positive control (Nicotinamide Mononucleotide (NMN), Merck Cat# N3501) in MEM medium (Himedia Cat# AL047S) supplemented with 1.9% Horse serum for 48 hrs. C2C 12 myotubes with 0.2% DMSO served as vehicle control. After the incubation, the medium was replaced with 50 pl of IX PBS and 50 pl of base solution (0.2N NaOH) with 1% Cetyltrimethylammonium bromide (CTAB; Sigma Cat# H5882) (Base treated sample). The plate was kept on the shaker briefly to ensure homogeneity and cell lysis. Protein was estimated using the Pierce BCA protein assay kit (Thermo Fisher Scientific Cat# 23225). After normalization, 50 pg protein lysate was added to all the wells of a 96-well flat- bottom plate for further steps.

[0131] NAD Measurement: NAD measurement was performed using the Promega NAD / NADH- Glo™ kit (Cat# G9071). Twenty -five microliters (containing 50 pg protein lysate) of each sample were added to empty wells in a 96-well flat-bottom plate for acid treatment. To these samples, 12.0pl of 0.4N HC1 was added (Acid treated sample). These wells contain acid-treated samples. The original sample wells are base-treated samples. The plate containing all samples was covered and incubated for 15 minutes at 60°C. After the incubation, the plate was equilibrated for 10 minutes at room temperature, and then 12.0 pl of 0.5M Trizma® base was added to each well of acid-treated cells to neutralize the acid. Twenty-five microliters of each sample were transferred into a 96-well, flat- bottom white plate (Corning Cat# 3912) for NAD measurement. Finally, 25 pl of NAD / NADH-Glo™ detection reagent was added and luminescence intensity was measured in a kinetic mode for upto 30 minutes. The percent increase of NAD over vehicle control as a result of sample treatment was calculated by the following formula:

[0132] % Increase in NAD over vehicle control = [(NAD concn, in sample-NAD concn, in vehicle control)] / NAD concn, in vehicle control x 100.

[0133] The results are presented in table 1.

[0134] Table 1: In vitro evaluation for NAD activation activity for various extracts of Punica granatum containing punicalagins, ellagic acid, and urolithin A

[0135] Example 10:

[0136] In vitro evaluation for CD38 inhibition activity for various extracts of Punica granatum containing punicalagins, ellagic acid, and urolithin A The L6 Rat skeletal myoblasts (ATCC Cat# CRL-1458) were maintained in 75cm2 cell culture flasks in DMEM medium (Himedia Cat# AL007S) supplemented with 10% FBS. Upon reaching 85% confluence, cells were washed with 1XPBS, trypsinized, and subjected to centrifugation at 250 x g for 10 minutes. The resultant cell pellet was washed with 1XPBS and was dissolved in 0.25 M Sucrose (Sigma Cat# S0389) buffer (containing 40mM of Tris base, pH 7.4) and stored at -80°C. The following day, the cell lysate was thawed, 1% TritonXIOO (Sigma Cat# T8787) was added and sonicated for 3 minutes with 1 minute ON: OFF cycle and centrifuged at 13500rpm for 10 minutes at 4°C. The resultant supernatant containing the CD38 enzyme was stored on ice until use or at -80°C for future use. Protein concentration was estimated using the Pierce BCA protein assay kit (Thermo Fisher Scientific Cat# 23225), and 25 pg / well protein was utilized for each reaction. In a 96-well Black round bottom polystyrene plate (Sigma Cat# CLS3792), 50 pl enzyme (equivalent to 25 pg protein), 50 pl test samples or positive control (Quercetin, Sigma Cat# Q4951) at different concentrations, and lOOpl (50pM) of Nicotinamide 1, N6- ethenoadenine dinucleotide (ENAD, Sigma Cat# N2630) were added. In parallel, enzyme control (only enzyme and ENAD) and buffer control wells (only buffer and ENAD) were also maintained. Relative Fluorescence Units (RFU) at endpoint was measured after Ihour of incubation in the dark at Ex / Em: 290 / 400nm in a Perkin Elmer EnSpire multimode plate reader (Waltham, MA). The %Inhibition of CD38 activity was calculated by the following formula:

[0137] %Inhibition of CD38 activity = [(Normalized RFU of Enzyme alone) - (Normalized RFU of Sample)] / Normalized RFU of Enzyme alone x 100

[0138] The results are presented in tables: 2.

[0139] Table 2: Percent inhibition of CD38 activity by various Punica granatum extracts containing punicalagins, ellagic acid, and Urolithin A

[0140] Example 11:

[0141] Preparation of Capsules:

[0142] General procedure: Ingredients 01 to 02 in a specified quantity were weighed and transferred into a Suitable mixer. The contents were mixed well and to this blend ingredients 03, 04 & 05 were added and the mass was mixed for 30-45 minutes. The blend was passed through 40 mesh sieve, and was used for filling in capsules.

[0143] TABLE 3

[0144] Capsule prepared from Punica granatum whole spectrum extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts as obtained from example 7.

[0145] Each capsule contains:

[0146] 01. Pomegranate extract of Example 7. 500.00mg

[0147] 02. Microcrystalline Cellulose . 50.00mg

[0148] 03. Sodium benzoate . 2.50mg

[0149] 04. Talcum . 2.50mg

[0150] 05. Magnesium stearate . 2.50mg

[0151] TABLE 4

[0152] Capsule prepared from Punica granatum whole spectrum extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts as obtained from example 8 along with other ingredients.

[0153] Each capsule contains: 01. Pomegranate extract of Example 8 ,250.00mg

[0154] 02. Curcuma longa rhizome extract ,100.00mg

[0155] 03. Bacopa monnieri whole plant extract lOO.OOmg

[0156] 04. Vitamin C 70.00mg

[0157] 05. Microcrystalline Cellulose 50.00mg

[0158] 06. Sodium benzoate 2.50mg

[0159] 07. Talcum 2.50mg

[0160] 08. Magnesium stearate 2.50mg

[0161] TABLE 5

[0162] Capsule prepared from Punica granatum whole spectrum extract from fresh fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts as obtained from example 7 along with other ingredients.

[0163] Each capsule contains:

[0164] 01. Pomegranate extract of Example 7 250.00mg

[0165] 02. Withania somnifera root extract lOO.OOmg

[0166] 03. Magnesium lOO.OOmg

[0167] 04. Vitamin C 50.00mg

[0168] 05. Vitamin D 25.00mcg

[0169] 06. Zinc 8.00mg

[0170] 07. Iron 5.00mg

[0171] 08. Microcrystalline Cellulose 50.00mg

[0172] 09. Sodium benzoate 2.50mg

[0173] 10. Talcum 2.50mg

[0174] 11. Magnesium stearate 2.50mg

[0175] TABLE 6

[0176] Capsule prepared from Punica granatum whole spectrum extract from fresh fruit containing dried rind, seeds containing arils, albedo, membrane and other fruit parts as obtained from example 3 along with other ingredients. Each capsule contains:

[0177] 01. Pomegranate extract of Example 3 250.00mg

[0178] 02. Terminalia arjuna bark extract lOO.OOmg

[0179] 03. Piper longum fruit extract 25.00mg

[0180] 04. Magnesium 30.00mg

[0181] 05. Vitamin K2 lOO.OOmcg

[0182] 06. Zinc 8.00mg

[0183] 07. Microcrystalline Cellulose 50.00mg

[0184] 08. Sodium benzoate 2.50mg

[0185] 09. Talcum 2.50mg

[0186] 10. Magnesium stearate 2.50mg

[0187] Example 12:

[0188] Compositions of Punica granatum whole spectrum extract from fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts along with other ingredients:

[0189] Encouraged by the increased efficacy in NAD boosting and CD38 inhibition activities of Punica granatum fruit as well as dried fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts as obtained from example 3, 4 and 7; and in combination with at least one ingredient selected from the extracts, fractions, phytochemicals or mixtures thereof derived from Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri, Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, , Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda.

[0190] Several compositions were prepared comprising, a Punica granatum whole spectrum extract obtained from fruit enriched with punicalagins, ellagic acid and urolithin A or mixtures thereof; and in combination with at least one ingredient selected from the extracts, fractions, phytochemicals or mixtures thereof derived from Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri, Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, , Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda. Further, Several compositions comprising, a Punica granatum whole spectrum extract obtained from fruit containing fruit rind, seeds containing arils, albedo, membrane and other fruit parts and in combination with at least one ingredient selected from vitamins are selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin B12, and other B vitamins, Vitamins C, Vitamin D and Vitamin K and the minerals are selected from the group consisting of Na, K, Mg, Ca and other essential minerals.

[0191] Thus, various solvent extracts of Curcuma longa and other Curcuma species, Withania somnifera, Bacopa monnieri, Asparagus racemosus, Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda were also prepared with different solvents.

[0192] Curcuma longa (rhizome), Withania somnifera (roots), Bacopa monnieri (whole plant), Asparagus racemosus (roots and / or leaves), Trigonella foenum granatum (seeds), Piper longum (fruits), Asparagus racemosus (roots) and Terminalia arjuna (bark) extracts: were pulverized and the powders were extracted separately with various solvents to obtain the corresponding extracts, and the extracts were standardized with their corresponding phytochemical markers using analytical HPLC method.

[0193] For study purpose, four compositions (comp # 1A, IB, 1C and ID) were prepared as follows:

[0194] (1 iC) Punica granatum whole spectrum extract as obtained from example 7, Curcuma longa rhizome extract and Piper longum fruit extract in the ratio of 5: 1 :4;

[0195] (IB) Punica granatum whole spectrum extract as obtained from example 7, Withania somnifera root extract and Piper longum fruits part extract in the ratio of 5:2:3;

[0196] (IC) Punica granatum whole spectrum extract as obtained from example 7, Asparagus racemosus (roots) extract and Bacopa monnieri whole plant part extract in the ratio of 4:4:2 and

[0197] (ID) Punica granatum whole spectrum extract as obtained from example 7, Asparagus racemosus roots extract and Terminalia arjuna bark extract in the ratio of 6:3 : 1.

[0198] These compositions were tested for their efficacy in boosting NAD levels and inhibiting CD38 in vitro cellular models, and the results are summarized in Table 2. Table 7: Efficacy of compositions comprising Punica granatum whole spectrum extract obtained from fruit containing fruit rind, seeds containing arils, albedo, membrane and in combination of other herbal extracts as well as essential vitamins & minerals

[0199] As per results obtained from above table, the composition-1 A comprising Punica granatum whole spectrum extract as obtained from example 7, Curcuma longa rhizome extract and Piper longum fruit extract in the ratio of 5: 1 :4 showed 38.78% increase in NAD level at 1 pg / mL, while 31.57% inhibition observed in CD38 levels. While the composition IB comprising Punica granatum whole spectrum extract as obtained from example 7, Withania somnifera root extract and Piper longum fruits part extract in the ratio of 5:2:3 at 1 pg / mL showed only 34.95% increase in NAD levels, while 23.79% inhibition was noted in CD38 levels. This increase in cellular NAD levels as well as CD38 levels of the inventive composition (composition- 1) is a surprising and synergistic result. Similarly, the composition- ID showed significantly better activation of NAD activity and composition- 1C showed significantly better inhibition of CD38 activity.

[0200] Example 13: Anecdotal Reports Subject 1:

[0201] Subject: 58- Year-Old Female Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0202] Duration: 3 Months

[0203] Background:

[0204] The subject, a 58-year-old female with a history of mild to moderate muscle stiffness and decreased mobility, began supplementation with pomegranate extract (500mg) capsules taken twice daily. Prior to starting supplementation, she reported experiencing daily stiffness, especially in the mornings and after periods of inactivity, leading to a reduction in overall mobility and physical activity.

[0205] Observations:

[0206] After 3 months of consistent supplementation, the subject reported noticeable improvements in muscle stiffness and overall mobility. She described feeling significantly "lighter" and more flexible, with a marked reduction in the stiffness she previously experienced. The subject also noted a consistent increase in her daily energy levels, which contributed to greater motivation and ability to engage in routine physical activities, such as walking and household chores, without the previous sense of fatigue or restriction.

[0207] Self-Reported Outcomes:

[0208] • Muscle Stiffness: Subject reported a substantial decrease in morning and postactivity stiffness.

[0209] • Mobility: Increased ease in walking, bending, and performing daily activities.

[0210] • Energy Levels: Noticeably higher, especially during the day, contributing to improved physical engagement and mood.

[0211] • Overall Wellbeing: Subject felt a sense of vitality and an improved outlook on physical activity.

[0212] Conclusion:

[0213] Over the course of three months, supplementation with pomegranate extract (500mg twice daily) appeared to positively impact the subject’s muscle stiffness, mobility, and energy levels. The subject attributes these improvements to the supplementation. Subject 2:

[0214] Subject: 69- Year-Old Male

[0215] Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0216] Duration: 3 Months

[0217] Background:

[0218] The subject, a 69-y ear-old male, reported experiencing frequent tiredness, reduced energy levels, and occasional lapses in cognitive sharpness, which had begun to affect his daily routines and overall quality of life. Seeking a natural approach to support his vitality, he commenced a regimen of pomegranate extract supplementation, taking 500mg twice daily.

[0219] Observations:

[0220] Following 3 months of consistent use, the subject noted significant improvements across several areas. He described feeling noticeably less tired throughout the day and reported enhanced stamina for completing daily tasks. His daily routines, including activities such as household management, personal exercise, and social engagement, became easier to maintain without the need for extended rest periods. In addition, he experienced improvements in cognitive functions, particularly in memory, focus, and mental clarity, which he had found to be gradually declining prior to supplementation.

[0221] Self-Reported Outcomes:

[0222] • Energy Levels: Marked increase in daily energy and reduced feelings of fatigue.

[0223] • Daily Routines: Improved ability to maintain consistent and active daily habits without exhaustion.

[0224] • Cognitive Function: Notable improvements in memory retention, mental clarity, and focus.

[0225] • Overall Wellbeing: Subject felt more engaged, motivated, and capable throughout the day.

[0226] Conclusion:

[0227] After three months of supplementation with pomegranate extract (500mg twice daily), the subject reported meaningful improvements in energy, endurance, daily functioning, and cognitive performance. These self-reported benefits suggest a positive role of pomegranate extract containing Urolithin A according to the present invention in supporting vitality and cognitive health in older adults.

[0228] Subject 3:

[0229] Subject: 71 -Year-Old Male

[0230] Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0231] Duration: 5 Months

[0232] Background:

[0233] The subject, a 71 -year-old male, initially reported persistent feelings of fatigue, gradual decline in hearing clarity, and weakened nail and hair health. To support his overall vitality, he began a supplementation regimen of pomegranate extract, taking 500mg twice daily.

[0234] Observations:

[0235] After 5 months of consistent supplementation, the subject reported significant positive changes. He experienced a notable reduction in feelings of fatigue, allowing him to maintain higher energy levels throughout the day. Additionally, he observed subtle but meaningful improvements in hearing clarity, including better recognition of conversations in moderate background noise. Furthermore, the subject reported visible enhancements in his hair and nail health, including stronger nails and thicker, healthier-looking hair.

[0236] Self-Reported Outcomes:

[0237] • Fatigue: Marked decrease in daily fatigue; greater endurance for physical and mental activities.

[0238] • Hearing: Improved clarity in hearing, particularly in social settings and environments with background noise.

[0239] • Nail and Hair Health: Stronger, faster-growing nails and visibly healthier, thicker hair.

[0240] • Overall Wellbeing: Subject expressed a renewed sense of vitality and confidence in daily life.

[0241] Conclusion:

[0242] Following five months of pomegranate extract supplementation (500mg twice daily), the subject reported notable improvements in fatigue levels, hearing clarity, and nail and hair health. These anecdotal results suggest a potential benefit of pomegranate extract containing Urolithin A in supporting aging-related health concerns.

[0243] Subject 4:

[0244] Subject: 51 -Year-Old Male

[0245] Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0246] Duration: 3 Months

[0247] Background:

[0248] The subject, a 51 -year-old male, reported experiencing fluctuations in mood and a decline in motivation for physical exercise, leading to reduced overall activity levels. Seeking a natural way to support his mental and physical wellbeing, he began supplementation with pomegranate extract, taking 500mg twice daily.

[0249] Observations:

[0250] After 3 months of consistent use, the subject noted significant improvements in both mood and physical performance. He reported feeling more emotionally balanced, with reduced periods of irritability and low motivation. Alongside these mental health improvements, he experienced greater enthusiasm and endurance for physical exercise, leading to more regular workout routines and improved physical stamina.

[0251] Self-Reported Outcomes:

[0252] • Mood: Improved emotional stability, reduced irritability, and enhanced overall sense of wellbeing.

[0253] • Exercise Performance: Increased motivation and stamina for regular physical exercise.

[0254] • Energy and Motivation: Higher energy levels and a greater willingness to engage in daily activities.

[0255] • Overall Wellbeing: Subject felt mentally and physically rejuvenated.

[0256] Conclusion:

[0257] Over three months of pomegranate extract supplementation (500mg twice daily), the subject experienced notable improvements in mood stability and exercise performance. These self-reported outcomes suggest potential benefits of pomegranate extract containing Urolithin A according to the present invention for supporting mental and physical health.

[0258] Subject 5:

[0259] Subject: 57-Year-Old Female

[0260] Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0261] Duration: 6 Months

[0262] Background:

[0263] The subject, a 57-y ear-old female, had been experiencing mood fluctuations, persistent fatigue, and a noticeable decline in muscle strength and endurance. In an effort to support her overall wellness, she began taking pomegranate extract at a dose of 500mg twice daily.

[0264] Observations:

[0265] After 6 months of consistent supplementation, the subject reported clear improvements. She described a more stable and positive mood, with fewer periods of emotional low and increased mental resilience. Additionally, she experienced a significant reduction in daily fatigue, allowing her to be more active and engaged in her daily routines. Improvements in muscle strength and endurance were also noted, with the subject reporting greater ease in performing physical tasks and longer-lasting energy during exercise and daily activities.

[0266] Self-Reported Outcomes:

[0267] • Mood: Increased emotional stability and positivity.

[0268] • Fatigue: Reduced feelings of tiredness, greater daily energy levels.

[0269] • Muscle Strength and Endurance: Enhanced physical strength and longer endurance during physical activities.

[0270] • Overall Wellbeing: Subject expressed a sense of rejuvenation and increased confidence in her physical capabilities.

[0271] Conclusion:

[0272] Following six months of pomegranate extract supplementation (500mg twice daily), with noticeable improvements beginning after three months, the subject reported better mood, reduced fatigue, and improved muscle strength and endurance. These self-reported benefits highlight the potential of pomegranate extract containing Urolithin A according to the present invention as a supportive supplement for physical and emotional health.

[0273] Subject 6:

[0274] Subject: 60-Year-Old Male

[0275] Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0276] Duration: 3 Months

[0277] Background:

[0278] The subject, a 60-y ear-old male, had been experiencing ongoing muscle pain and discomfort, which affected his daily activities and overall quality of life. Looking for a natural way to manage these symptoms, he began a supplementation routine with pomegranate extract, taking 500mg twice daily.

[0279] Observations:

[0280] After 3 months of regular supplementation, the subject reported a noticeable reduction in muscle pain and general discomfort. He described feeling less stiffness and soreness, particularly after periods of physical activity. This improvement allowed him to move more freely and engage more comfortably in his everyday tasks without the usual level of muscular discomfort he had been experiencing.

[0281] Self-Reported Outcomes:

[0282] • Muscle Pain: Significant reduction in muscle soreness and tenderness.

[0283] • Discomfort: Decreased general body discomfort, especially after activity.

[0284] • Mobility and Activity: Improved ease of movement and willingness to stay physically active.

[0285] • Overall Wellbeing: Increased physical comfort contributed to a better sense of daily wellbeing.

[0286] Conclusion:

[0287] After three months of consistent pomegranate extract supplementation (500mg twice daily), the subject reported meaningful relief from muscle pain and discomfort. These selfreported improvements suggest that pomegranate extract containing Urolithin A according to the present invention may offer supportive benefits for muscle health and comfort. Subject 7:

[0288] Subject: 55-Year-Old Male

[0289] Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0290] Duration: 5 Months

[0291] Background:

[0292] The subject, a 55 -year-old male, reported experiencing a gradual decline in physical energy and performance, making it more difficult to maintain his usual level of physical activity. To address this, he began a supplementation regimen with pomegranate extract, taking 500mg twice daily.

[0293] Observations:

[0294] After 5 months of consistent supplementation, the subject noted significant improvements in his physical energy and ability to engage in physical activities. He reported feeling more energized throughout the day, with greater stamina during exercise and daily tasks. Activities that previously felt tiring became easier to manage, and his overall physical endurance noticeably improved.

[0295] Self-Reported Outcomes:

[0296] • Physical Activities: Increased ability to perform and sustain regular physical activities.

[0297] • Energy Levels: Marked improvement in daily energy and reduced fatigue.

[0298] • Endurance: Enhanced stamina during both moderate and more strenuous activities.

[0299] • Overall Wellbeing: Improved sense of vitality and physical confidence.

[0300] Conclusion:

[0301] Following five months of pomegranate extract supplementation (500mg twice daily), the subject experienced substantial benefits in physical energy and activity levels. These selfreported outcomes suggest that pomegranate extract containing Urolithin A according to the present invention may be supportive of energy enhancement and physical performance.

[0302] Subject 8:

[0303] Subject: 73 -Year-Old Male Supplement: 500mg Pomegranate extract of Example 7, Twice Daily

[0304] Duration: 5 Months

[0305] Background:

[0306] The subject, a 73 -year-old male, had been experiencing reduced physical stamina, along with noticeable changes in skin firmness and hair health commonly associated with aging. In an effort to support his overall vitality and appearance, he began taking pomegranate extract, 500mg twice daily.

[0307] Observations:

[0308] After 5 months of consistent supplementation, the subject reported significant improvements across several areas. He experienced enhanced physical activity levels, with greater ease and endurance during exercise and daily movements. In addition, he noticed improved skin firmness, with his skin feeling more resilient and looking healthier. He also observed better hair health, including improved texture and thickness, and reduced hair thinning.

[0309] Self-Reported Outcomes:

[0310] • Physical Activities: Increased stamina and ease during exercise and daily tasks.

[0311] • Skin Firmness: Improved skin texture, firmness, and overall appearance.

[0312] • Hair Health: Healthier, thicker hair with less noticeable thinning.

[0313] • Overall Wellbeing: A greater sense of vitality, physical confidence, and wellbeing.

[0314] Conclusion:

[0315] Following five months of pomegranate extract supplementation (500mg twice daily), the subject reported significant improvements in physical activity levels, skin firmness, and hair health. These positive self-reported effects suggest that pomegranate extract containing Urolithin A according to the present invention may contribute to enhanced physical vitality and support healthy aging.

[0316] Subject 9:

[0317] Subject: 70-Year-Old Female

[0318] Supplement: Pomegranate Extract 500mg, Twice Daily Duration: 3 Months

[0319] Background:

[0320] The subject, a 70-year-old female, had been experiencing frequent episodes of brain fog, reduced mental clarity, and muscle stiffness, which impacted her daily activities and overall quality of life. To support her cognitive and physical wellbeing, she began a supplementation regimen with pomegranate extract, taking 500mg twice daily.

[0321] Observations:

[0322] After 3 months of consistent supplementation, the subject reported notable improvements. She experienced a significant reduction in brain fog, with clearer thinking, better focus, and improved memory during daily tasks. Alongside the cognitive benefits, she also observed a decrease in muscle stiffness, allowing her greater flexibility and ease of movement throughout the day.

[0323] Self-Reported Outcomes:

[0324] • Mental Clarity: Improved focus, sharper thinking, and reduced episodes of brain fog.

[0325] • Muscle Stiffness: Noticeable reduction in stiffness, leading to improved mobility.

[0326] • Daily Activities: Increased ease in managing both mental and physical tasks.

[0327] • Overall Wellbeing: Enhanced sense of mental alertness and physical comfort.

[0328] Conclusion:

[0329] Following three months of pomegranate extract supplementation (500mg twice daily), the subject reported improvements in both cognitive function and muscle flexibility. These selfreported benefits suggest that pomegranate extract containing Urolithin A according to the present invention may play a supportive role in promoting mental clarity and physical comfort, particularly in older adults.

Claims

im,1. Compositions of Punica granatum whole spectrum extract (PWSE) extract comprising Urolithin A in the range of 0.2% - 10%, for improving general health, overall wellbeing and longevity, by preventing age related disorders.

2. The compositions of Punica granatum whole spectrum extract (PWSE) as claimed in claim 1, wherein the extract is selectively enriched to comprise Punicagalins; Ellagic acid; and Urolethin A.

3. The compositions as claimed in claim 2, wherein, the Punica granatum whole spectrum extract (PWSE) selectively enriched to comprises a) Punicagalins in the range of 2.0% - 35.0%; b) Ellagic acid in the range of 0.5% - 14.0%; and c) Urolethin A in the range of 0.2% - 10.0%.

4. The compositions as claimed in any one of the claims 1 to 3 wherein, the Punica granatum whole spectrum extract (PWSE) comprises a) Punicagalins in the range of 2.0% - 35.0%; b) Ellagic acid in the range of 0.5% - 14.0%; c) Urolethin Ain the range of 0.2% - 10.0%; d) Galic acid in the range of 0.1% - 2.5%; e) Tannins in the range of 5.0% - 50.0%; and f) other Polyphenols in the range of 1.0% - 15.0%.

5. The compositions as claimed in any one of the claims 1 to 4, wherein, the compositions comprise; a) Punica granatum whole spectrum extract (PWSE) selectively enriched with Punicagalins, Ellagic acid and Urolethin A; b) at least one herb / herbal extracts selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera. Bacopa monnieri. Asparagus racemosus. Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda; and c) optionally fortified with one or more vitamins and minerals.

6. The composition as claimed in claim 5, wherein, the vitamins are selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin Bl 2, and other B vitamins, Vitamins C, Vitamin D and Vitamin K and the minerals are selected from the group consisting of Na, K, Mg, Ca and other essential minerals.

7. The composition as claimed in any one of the claims 1 to 6, wherein, the composition may be formulated into a dosage form selected from powder, tablet, acapsule, a soft chewable tablet or liquid form, beverage, food product, dietary supplement, or any other suitable forms using suitable conventional pharmaceuti cal / nutraceuti cal excipi ents .

8. The composition as claimed in claim 1, wherein, the age related disorders are selected from the group consisting of impaired cognitive functions including neurodegenerative disorders with symptoms of memory loss, cognitive decline, motor dysfunction, neuroinflammation, musculoskeletal degeneration; mitochondrial diseases with symptoms of muscle fatigue, exercise intolerance, organ dysfunction and metabolic disorders with symptoms of insulin resistance, abnormal fat accumulation, fatigue as well as other age related common symptoms such as mood disturbances, muscle weakness, poor sleep, and slower recovery from illness.

9. A process for preparation of Punica granatum whole spectrum extract (PWSE / or PWSpE) selectively enriched to comprise Punicagalins; Ellagic acid; and Urol ethin A, comprising a step of extracting the Punica granatum whole fruit with a source of probiotic.

10. The process as claimed in claim 9, wherein, the source of probiotic is selected from the group consisting of milk, yogurt and a probiotic blend.

11. The process as claimed in claim 9, wherein, the Punica granatum whole fruit include the fruit parts such as fruit rind, seeds containing arils, albedo, membrane and other fruit parts of Punica granatum.

12. The process as claimed in claim 9, wherein, the process for preparation of Punica granatum whole spectrum extract (PWSE) comprising the steps of; a) extracting the Punica granatum whole fruit with a solution containing mixture of milk and water for 6-10 h with intermittent stirring at ambient temperature; b) filtering the liquid extract from the remaining fruit mass and isolating the Punica granatum milk extract powder (PME) by evaporating the liquid extract; c) subjecting the remaining fruit mass to repeated aqueous extractions at a temperature of 90-95°C; d) filtering the aqueous extract from the remaining fruit mass;e) subjecting the remaining fruit mass to repeated alcohol extractions at a temperature of 60-65°C; f) filtering the alcohol extract from the remaining fruit mass, combining the aqueous and alcohol extracts and evaporating the extract to isolate the Punica granatum extract powder enriched with high polarity phytoconstituents-PEHPP; and g) combining the Punica granatum milk extract powder of step b) and Punica granatum extract powder enriched with high polarity phytoconstituents- PEHPP of step f) to obtain Punica granatum whole spectrum extract (PWSE).

13. The process as claimed in claim 9, wherein, the process for preparation of Punica granatum whole spectrum extract (PWSpE) extracted with a proprietary probiotic blend, comprising the steps of; a) extracting the Punica granatum whole fruit with a solution containing a mixture of the probiotic blend and water for 16-20 h with intermittent stirring at ambient temperature; b) filtering the liquid extract from the remaining fruit mass and isolating the Punica granatum probiotic extract powder (PPE) by evaporating the liquid extract; c) subjecting the remaining fruit mass to repeated aqueous extractions at a temperature of 90-95°C; d) filtering the aqueous extract from the remaining fruit mass; e) subjecting the remaining fruit mass to repeated alcohol extractions at a temperature of 60-65°C; f) filtering the alcohol extract from the remaining fruit mass, combining the aqueous and alcohol extracts, evaporating the combined extract to isolate the Punica granatum extract powder post probiotic treatment- PEPPT; and g) combining the Punica granatum probiotic extract powder (PPE) of step b) and Punica granatum extract powder post probiotic treatment - PEPPT of step f) to obtain Punica granatum whole spectrum extract (PWSpE).

14. The process as claimed in claim 9 or claim 13, wherein, the probiotic blend is a proprietary blend that consisting of Lactobacillus acidophilus, Bifidobacteriumlactis, Bifidobacterium infantis, Lactobacillus plantarum, L. salivarius and B. longum.

15. A method of increasing NAD levels in a human subject, which method comprises supplementing the human subject in need thereof, a therapeutically effective amounts of a composition comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10% as claimed in claims 1 to 5, and optionally at least one ingredient selected from a herb / herbal extract, vitamins, minerals, pharmaceutically or nutraceutically or dietically acceptable excipients, carriers, and diluents.

16. A method of inhibiting CD38 levels in a human subject, which method comprises supplementing the human subject in need thereof, a therapeutically effective amounts of a composition comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, as claimed in claims 1 to 5 and optionally at least one ingredient selected from the group consisting of a herb / herbal extract, vitamins, minerals, pharmaceutically or nutraceutically or dietically acceptable excipients, carriers, and diluents.

17. The method as claimed in claim 15 or 16, wherein the herb / herbal extract is selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera. Bacopa monnieri. Asparagus racemosus. Trigonella foenum granatum, Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda.

18. The method as claimed in claim 15 or 16, wherein the vitamins are selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin B 12, and other B vitamins, Vitamins C, Vitamin D and Vitamin K and the minerals are selected from the group consisting of Na, K, Mg, Ca and other essential minerals.

19. Use of compositions comprising Punica granatum whole spectrum extract selectively enriched to comprise Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, and optionally at least one ingredient selected from an herb / herbal extract, vitamins, minerals, pharmaceutically or nutraceutically or dietically acceptable excipients, carriers, and diluents, for increasing NAD levels in a human subject.

20. Use of compositions comprising Punica granatum whole spectrum extract selectively enriched to contain Punicagalins in the range of 2% -35%; Ellagic acid in the range of 0.5% - 14% along with Urolethin A in the range of 0.2% - 10%, and optionally at least one ingredient selected from the group consisting of an herb / herbal extract, vitamins, minerals, pharmaceutically or nutraceutically or dietically acceptable excipients, carriers, and diluents, for inhibiting CD38 levels in a human subject.

21. The use as claimed in claim 19 or 20, wherein the herb / herbal extract is selected from the group consisting of Curcuma longa and other Curcuma species, Withania somnifera. Bacopa monnieri. Asparagus racemosus. Trigonella foenum granatum,Piper longum and other Piper species, Terminalia arjuna and other Terminalia species and other herbs described as “Rasayana” in Ayurveda.

22. The use as claimed in claim 19 or 20, wherein the vitamins are selected from the group consisting of Vitamin A, Vitamin B5, Vitamin B6, Vitamin Bl 2, and other B vitamins, Vitamins C, Vitamin D and Vitamin K and the minerals are selected from the group consisting of Na, K, Mg, Ca and other essential minerals.

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