DESC:A SYNERGISTIC HERBAL COMPOSITIONIMPROVING FOR MALE SEXUAL WELLNESS
FIELD OF THE INVENTION
The present invention relates to an oral synergistic herbal composition that improves sexual health and cellular functions in males. More specifically, the present invention relates to a synergistic herbal composition having the combination of herbs Tribulus terrestris and Panax ginseng.
BACKGROUND OF THE INVENTION
The sexual response cycle in both men and women is a complex interplay between the autonomic nervous system, endocrine system and circulatory system, that involves a plethora of biochemical factors like neurotransmitters, vasoactive agents and endocrine factors etc. It is well described in the scientific and medical literature that subjects having low levels of sex hormones are more likely to be depressed than those with normal hormonal levels. They also suffer from complex clinical outcomes likeloss of libido, erectile dysfunction, premature ejaculation, delayed or inhibited orgasm, physical abnormalities in penis etc. A multitude of pathways operates in a well-orchestrated manner in every stage of sexual responsive cycle such asdesire (libido), excitation (arousal), orgasm and resolution. To address the various problems of the sexual response cycle, the availability of drugs is scarce.Phosphodiesterase (PDE5) inhibitors for erectile dysfunction (ED), flibanserin(serotonin receptor modulator)for hypoactive sexual desire disorder (HSDD) are a few drugs available in the market todate. Furthermore, hundreds of productsexist in the market to address virility and erectile dysfunction (ED).
There are supplements available in the market that are purported to alleviate human sexual problems. But they are marketed with little scientific evidence and rely on popular belief and personal testimonials. Companies are focusing on investing towards the discovery of new ingredients that fall primarily into 3 main functional categories: testosterone boosters, PDE-5 inhibitors (e.g. sildenafil like activity), and stimulators of nitric oxide (NO) levels. Several types of researches have been carried out and are documented as patents of which few are listed here.
EP0714300 describes improved methods for modulating the human sexual response by administering a vasodilator by a route of administration that avoids the 'first-pass' effect.
CN102612370B relates to dietary supplements comprising Tribulus terrestris, glucosamine oligosaccharide and alga Ecklonia comprising fat-soluble polyphenols. The dietary supplements are useful in the prevention and/or the treatment of erectile dysfunction.
The lack of mainstream medication and various other reasons including side effects and ineffectiveness in certain populations has led to the growing popularity of Dietary supplements (DS) or Nutraceuticals for better management of sexual problems.Thus, the need for the present invention is to overcome the problems associated with the existing prior art.
OBJECTIVE OF THE INVENTION
The main objective of the present invention is to develop a synergistic herbal combination that up-regulates sex hormones thereby involve sexual health and cellular functions in males.
Yet another objective of the present invention is to provide a palatable hence easy to consume beverage despite being a herbal composition.
Yet another objective of the present invention is to provide a scientifically validated and functional nutraceutical composition.
Yet another objective of the present invention is to provide a synergistic herbal composition having an extended shelf life.
Yet another objective of the present invention is to provide a synergistic herbal composition that improves libido and helps maintain sexual balance.
Further objectives, advantages, and features of the present invention will become apparent from the detailed description provided hereinbelow, in which various embodiments of the disclosed invention are illustrated by way of example.

SUMMARY OF THE INVENTION
The present invention describes as herbal sexual wellness formulation for males providing synergistic effects in-vitro. The herbal sexual wellness formulation comprising of an extract of Ingredient A and ingredient B used in the concentration range of 0.001 – 500 µg/ml. Herein, the ingredient A is an extract from the fruit of the plant but not limited to stem, leaf, inflorescence, or root extract, and the ingredient B is an extract from the root of the plant but not limited to fruit, leaf, stem, seed or inflorescence extract. Herein, the herbal extract is an aqueous extract but not limited to ethanol, methanol, acetone, ethyl acetate extracts and contains the bio-actives protodioscin, alkaloids, steroidal saponins, furostanol saponins, flavonoid glycosides, ginsenosides. In an embodiment, the ingredient A is obtained from Tribulus terrestris but not limited to T.terrestris orientalis, T.terrestris var robustus, T.terrestris var brachyceras,T.terrestris var inermis,T. terrestris var sericeus, T. terrestris var rajasthanensis, T.terrestris var macrocarpus, T.terrestris var desertorum, T.terrestris var biocornutus and the ingredient B is obtained from Panax ginseng but not limited to Panax quinquefolius var ginseng, P. notoginseng, Panax ginseng var. repens. In an embodiment of the present invention, the herbal formulation has been evaluated in vitro using cell lines, wherein the cell lines can be of mammalian, avian, Piscean, or arthropod origin. Herein, the cell type is MA-10 cells but not limited to other interstitial testicular cell lines. In an embodiment of the present invention, at least one pharmaceutically accepted excipient is selected from, but not limited to, coloring agents such as allura red and flavoring agents such as strawberry powder, acidity regulator such as sodium citrate and citric acid but not limited to acetic acid, malic acid, lactic acid, adipic acid and other salts, preservative such as sodium benzoate but not limited to other similar preservatives, emulsifier/stabilizer/gelling agent/ thickener such as xanthan gum but not limited to other similar agents, added in the range 0.05 to 0.5 w/v%. The present invention is an herbal formulation administered orally and is selected from, but not limited to, a pill, tablet, capsule, concentrated syrup, food additive, suspension, emulsion, powdered drink mix, carbonated/non-carbonated beverage and a chewable solid. In an aspect of the present invention, the in vitro evaluation is done using the specific combination of ingredient A and ingredient B taken in a concentration range of 0.0001–50 µg/ 104 MA-10 cells, has synergistic effects on upregulation of male sex hormone testosterone and influence on libido and sexual health in males. In another aspect of the present invention, the specific combination of Ingredient A and Ingredient B provide synergistic effects by modulating nitric oxide levels, which in turn influences intracellular calcium levels. This leads to phosphorylation/dephosphorylation of myosin light chain thereby influencing penile erection through smooth muscle relaxation.
The main advantage of the present invention is to develop a synergistic herbal combination that up-regulates sex hormones thereby improves sexual health and cellular functions in males.
Yet another advantage of the present invention is that the present invention provides a palatable and easy to consume herbal composition.
Yet another advantage of the present invention is that the present invention provides a scientifically validated functional nutraceutical composition.
Yet another advantage of the present invention is that the present invention provides a synergistic herbal composition having an extended shelf life.
Yet another advantage of the present invention is that the present invention provides a synergistic herbal composition that improves libido and helps maintain sexual balance.
Further objectives, advantages, and features of the present invention will become apparent from the detailed description provided herein below, in which various embodiments of the disclosed invention are illustrated by way of example..

DETAILED DESCRIPTION OF THE INVENTION
Definition

The term “a” or “an”, as used herein, is defined as one. The term “plurality”, as used herein, is defined as two as or more than one. The term “another”, as used herein, is defined as at least a second or more. The terms “including” and/or “having”, as used herein, are defined as comprising (i.e., open language).
The term “comprising” is not intended to limit the present invention with such terminology rather is used in a wider sense. Any invention using the term comprising could be separated into one or more claims using “consisting” or“consisting of”. The term “comprising” may be used interchangeably with the terms “having” or “containing”.
Reference in this document to “one embodiment”, “certain embodiments”, “an embodiment”, “another embodiment”, and “yet another embodiment” or similar terms, throughout the document means that a specific feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the appearances of such phrases in various places, this specification throughout are not necessarily all referring to the same embodiment. Furthermore, the specific features, structures, or characteristics are combined in any suitable manner in one or more embodiments without limitation.
The term “or” as used herein is to be interpreted as inclusive or meaning any one or more combinations. Therefore, “A, B or C” means any of the following: “A; B; C; A and B; A and C; B and C; A, B and C”. An exception to this definition will occur only when a combination of elements, functions, steps, or acts are in mutually exclusive, inherently.
As used herein, the term "one or more" generally refers to, but is not limited to, singular as well as the plural form of the term.
The drawings featured in the figures are to illustrate certain convenient embodiments of the present invention and are not to be considered as a limitation to that. Term "means" preceding a present participle of operation indicates the desired function for which there are one or more embodiments, i.e., one or more methods, devices, or apparatuses for achieving the desired function and that one skilled in the art could select from these or their equivalent in view of the disclosure herein and use of the term "means" is not intended to be limiting.
Fig.1 illustrates MTT assay. 1ng to 1000µg/ml of both T.terrestris and P.ginseng were incubated with 8 x 104 MA10 cells for 24 hrs and MTT performed at the end of 24hrs incubation

Figure 2 .illustrates Dose dependent increase in testosterone with herbal combination. A. Standard curve obtained with the testosterone standard provided with Cayman kit. X axis indicates the concentrations of standards used. Y-axis indicates the Logit (B/B0) values obtained with the excel worksheet provided by Cayman for calculation of Testosterone. B. Dose dependent increase in concentration of testoserone estimated from culture supernatant after incubation of herbs with MA10 cells for 24hrs. Herb A-P.ginseng. Herb B-T.terrestris. Herb A at 500µg/ml produces 822pg/ml testosterone, which increases to 860, 874, 917, 922 and 999pg/ml with additive effect of 0.001, 0.01, 10, 100 and 500µg/ml of Herb B respectively.

Figure 3. illustrates Dose dependent increase in Nitrate with herbal combination. Nitrate/Nitrites are byproducts of Nitric oxide that is generated intracellularly during the upregulation of NO/cGMP signalling. A. Standard curve obtained with the Nitrate standard. X axis indicates the concentrations (uM) of standards used. Y-axis indicates the OD at 540nm. B. Dose dependent increase in concentration of Nitrate/nitrite estimated from culture supernatant after incubation of herbs with MA10 cells for 24hrs. Herb A-P.ginseng. Herb B- T.terrestris. T.terrestris at 500µg/ml produces 12uM nitrite/nitrate, which increases to 13.7, 14.9 and 15.6 uM/ml with additive effect of 10, 100 and 500µg of P.ginseng respectively

Figure 4. illustrates Box & Whisker Plot for % improvement in the Score of MSQ from Visit 1 to Visit 3 by Two Treatment Arms – Full Analysis Set. Total score of Subject Investigational product Feedback Questionnaire at the end of the 2 month study compared between 2 treatment arms: (1) Test arm (2) placebo- control arms

Figure 5. illustrates Improvement in MSQ score comprising of 5 questions over 2 month study period. Te 5 questions for which, the score are shown individually are: 1. Desire to Initiate Sexual Intercourse (i.e., Desire) 2. Foreplay enjoyable (i.e., Foreplay) 3. Maintain Erection (i.e., Maintain Erection) 4. Control Ejaculation (i.e., Control Ejaculation) 5. Sexual Performance (i.e., Sexual Performance).

Figure 6 illustrates Schematic representation of Clinical trial- Study design

Figure 7 illustrates Composition of drinks used for Clinical trial. Clinical efficacy of Male sexual wellness was evaluated against placebo control drink that had all other ingredients except the herbs that act synergistically.

The present invention describes as herbal sexual wellness formulation for males providing synergistic effects in-vitro. The herbal sexual wellness formulation comprising of an extract of Ingredient A and ingredient B used in the concentration range of 0.001 – 500 µg/ml. Herein, the ingredient A is an extract from the fruit of the plant but not limited to stem, leaf, inflorescence, or root extract, and the ingredient B is an extract from the root of the plant but not limited to fruit, leaf, stem, seed or inflorescence extract. Herein, the herbal extract is an aqueous extract but not limited to ethanol, methanol, acetone, ethyl acetate extracts and contains the bio-actives protodioscin, alkaloids, steroidal saponins, furostanol saponins, flavonoid glycosides, ginsenosides. In an embodiment, the ingredient A is obtained from Tribulus terrestris but not limited to T.terrestris orientalis, T.terrestris var robustus, T.terrestris var brachyceras,T.terrestris var inermis,T. terrestris var sericeus, T. terrestris var rajasthanensis, T.terrestris var macrocarpus, T.terrestris var desertorum, T.terrestris var biocornutus and the ingredient B is obtained from Panax ginseng but not limited to Panax quinquefolius var ginseng, P. notoginseng, Panax ginseng var. repens. In an embodiment of the present invention, the herbal formulation has been evaluated in vitro using cell lines, wherein the cell lines can be of mammalian, avian, Piscean, or arthropod origin. Herein, the cell type is MA-10 cells but not limited to other interstitial testicular cell lines. In an embodiment of the present invention, at least one pharmaceutically accepted excipient is selected from, but not limited to, coloring agents such as allura red and flavoring agents such as strawberry powder, acidity regulator such as sodium citrate and citric acid but not limited to acetic acid, malic acid, lactic acid, adipic acid and other salts, preservative such as sodium benzoate but not limited to other similar preservatives, emulsifier/stabilizer/gelling agent/ thickener such as xanthan gum but not limited to other similar agents, added in the range 0.05 to 0.5 w/v%. The present invention is an herbal formulation administered orally and is selected from, but not limited to, a pill, tablet, capsule, concentrated syrup, food additive, suspension, emulsion, powdered drink mix, carbonated/non-carbonated beverage and a chewable solid. In an aspect of the present invention, the in vitro evaluation is done using the specific combination of ingredient A and ingredient B taken in a concentration range of 0.0001–50 µg/ 104 MA-10 cells, has synergistic effects on upregulation of male sex hormone testosterone and influence on libido and sexual health in males. In another aspect of the present invention, the specific combination of Ingredient A and Ingredient B provide synergistic effects by modulating nitric oxide levels, which in turn influences intracellular calcium levels. This leads to phosphorylation/dephosphorylation of myosin light chain thereby influencing penile erection through smooth muscle relaxation.

Tribulus terrestrisis from the caltrop family. It is known as puncture vine or Gokharu. The fruit is used as traditional Chinese medicine. It is known to be effective in increasing testosterone levels. In addition, Protodioscin is a steroidal saponin from Tribulus terrestis plant that has been postulated to mimic or increase the levels of DHEA (Di-Hydro-Epi-Androsterone). A study conducted by Do et.al., proved the efficacy of T.terrestris extract on phenylephrine (contractile agent) pretreated corpora cavernosa tissue isolated from rabbits. The results showed an increase in relaxation of the tissue in a dose-dependent fashion. cAMP levels were also found to be elevated at all doses of T.terrestris administration. This study shows that T.terrestris helps in mediating NO signaling pathway and its constituent regulators such as cAMP.
Panax ginseng has been used as a traditional Chinese medicine to enhance sexual performance and satisfaction. The potential effect of P.ginseng is mainly dependent on ginsenosides. Among the ginsenosides, Rg1 has been found to influence NO production in endothelial cells. Wang et.al., studied the effect of Rg1 on the copulatory behavior in male mice and NO/cGMP pathway in erectile tissue both in vivo and invitro. However, no studies exist till date to study the combinatorial effects of the herbs. The present invention addresses how the herbal combinations act synergistically in this aspect. The present invention also helps to improve and promote strength, vigor and maintain healthy sexual life in human males.
The present invention claims an increase in both NOS and testosterone activity using the synergistic combination of extracts of Tribulus terrestris and Panax Ginseng. The effect of these extracts on male sexual wellness has been validated using leydig tumor cell line. The scientific efficacy evaluated in cell lines were substantiated with a 2 month double blinded clinical trial conducted in healthy men volunteers and outcome measured at the end of study period. The clinical trial outcomes have shown promising results and the current thus strongly prove the efficacy of the herbal combination in effectively improving sexual functions in men.
In an embodiment of the present invention, the herbal sexual wellness formulation for providing synergistic combination of aqueous extracts of Panax ginseng and Tribulus terrestris in the ratio 1:1 to 1:5x105 effects in vitro cell based study comprise of confer health benefits related to sexual and other illness in men. The synergistic herbal combination as mentioned, wherein T.terrestris and Panax ginseng in the ratio of 1:1 to 1:5x105 .The synergistic herbal combination as mentioned , wherein Panax ginseng and Tribulus terrestris upregulated the synthesis of male sex hormone Testosterone as estimated from the culture supernatant after 24hrs of incubation. The synergistic herbal combination as mentioned, wherein Panax ginseng and Tribulus terrestris upregulated the production of cellular Nitric oxide as estimated from the culture supernatant after 24hrs of incubation.

Materials and methods:
Cell culture
MA10 is a Murine Leydig cell line that is obtained from ATCC and was cultured in MEM:F12 (1:1) supplemented with 15% horse serum (HS). Cell lines were cultivated in the presence of benzyl penicillin and streptomycin and were maintained at 37o C in a humidified atmosphere of 5% CO2
Herbal extracts
T.terrestris aqueous dry extracts of whole fruit (extract of 10:1) and P.ginseng aqueous root extract (10% Ginsenosides) were purchased from Herbo Nutra, India.
Cell viability assessment – MTT assay
MTT (Dimethylthiazol-diphenyltetrazolium bromide) is a colorimetric assay that assess cell viability by determining these mitochondrial functional state. Mitochondria in a living cell secretes NAD(P)H-oxidoreductases which reduces yellow coloured MTT taken up by cells due to its net positive charge to form violet coloured formazan crystals. For this study, MA10 cells were seeded in 96 well plate at a concentration of 8 x 104 cells/well (100ul cell suspension) and incubated for 24 hours. Herbal extracts ranging from 0.5mg/ml to 10ng/ml concentration were added after the incubation. MTT assays were performed for 2hr, 4hr, 8hr and 24 hrs (only 24hrs read out depiced in this patent), by adding 10µl of 5mg/ml MTT reagent and was incubated at 37o C in dark for 2hrs. After the incubation time, 100µl of crystal dissolving solution (solution containing 20% SDS and 50% DMF) was added and then incubated in dark until the crystals were dissolved. OD was measured at 560nm, cell viability was calculated as follows:

Nitric oxide estimation using Griess reagent:
MA10 cells were seeded at a density of 8 X 104 cells /well. At the end of 24-hour incubation, the cells are treated with various concentration of herbs in specific combinations. After 24hrs or incubation, the total nitrate/nitrite in the cell supernatant is estimated using colorimetric assay kit (cat no: 760871) Cayman chemicals (USA). Average OD values from 3 separate assays are subtracted from the blank values. Standard curve is plotted using Nitrate/Nitrite standards (1-25 µM). Concentration of nitrate/nitrite in the samples was determined using the standard curve.
Competitive ELISA for estimation of Testosterone:
Cells were seeded at a density of 8 X 104 cells /well. At the end of 24-hour incubation, the cells are treated with various concentrations of herbs. After 24hrs of incubation, Testosterone in the culture supernatant of the MA10 cells were estimated with competitive ELISA kits obtained from Cayman chemicals (USA) (cat no 58271). B/B0 values are calculated for all the samples (B indicates ‘% bound’ i.e., percentage of analyte that competes with the given tracer in the sample. B0 indicates ‘maximum bound’ i.e., the value obtained with the maximum binding of tracer in the absence of analyte). With the B/B0 values obtained for standards, the 4-parameter logistic fit standard curve is plotted as mentioned in the kit. Samples concentrations were estimated using the equation obtained from the standard curve.
Statistical analysis:
All data are represented as mean ±standard deviation (SD). Differences between the groups were determined with Analysis of variance (ANOVA) followed by the LSD multiple comparison tests, using SPSS version 25. The data is considered statistically significant if P=0.05
Double blinded, prospective, randomized, comparative, placebo controlled clinical trial:
Design:
This clinical study was designed a double blinded, prospective, randomized, comparative, placebo controlled clinical study to evaluate the effectiveness, safety and tolerability of Male Sexual Wellness Drink formulated with unique formula with scientifically tested ingredients in improving Sexual Wellness in Healthy Adult Male Subjects. Healthy adult men between 21 and 65 years of age who were in a heterosexual, monogamous relationship with an active sexual life were screened for the clinical study. Apart from the inclusion exclusion criteria, the subjects were also screened using the Male Sexual Quotient Questionnaire (MSQ).
Treatment arms:
Subjects who were eligible were enrolled into the study and randomized into two treatment arms:
Treatment Arm I : 33 Subjects : Male Sexual Wellness Drink
Treatment Arm II : 11 Subjects : Placebo
Subjects consumed 2 drinks/day (1-ounce liquid shots) for a period of 2 months and MSQ analysis done (a) at the time of induction (b) end of 1 month and (c) end of 2 months study period

Outcome measure:
Efficacy was analysed using Male Sexual Quotient Questionnaire (MSQ) which is a standardized questionnaire to evaluate the improvement in sexual wellness in adult males.
There are five questions considered un this outcome measure MSQ and those questions are related to the following five attributes:
1. Desire to Initiate Sexual Intercourse (i.e., Desire)
2. Foreplay enjoyable (i.e., Foreplay)
3. Maintain Erection (i.e., Maintain Erection)
4. Control Ejaculation (i.e., Control Ejaculation)
5. Sexual Performance (i.e., Sexual Performance)
Null hypothesis, H0: Mean % improvement in MSQ-5Q by Treatment Arm-I is not significantly different from that of Treatment Arm-II (i.e., µ1 = µ2) Alternate hypothesis, H1: Mean % improvement in MSQ-5Q by Treatment Arm-I is significantly greater than that of Treatment Arm-II (i.e., µ1 > µ2) Statistical Test to be applied: Two Mean Test or Independent Samples t-test
Statistical analysis:
Per the statistical analysis plan, 44 subjects were enrolled so as achieve an evaluable total of 40subjects. All statistical analysis is performed in accordance with the ICH E9 guideline for Statistical Principles for Clinical Trials, using SAS® (Version 9.4 or higher). The efficacy of each individual treatment arm was determined by Single Proportion Test and comparative efficacy for proportion of subjects was determined by Two Proportion Test. For absolute scores, the efficacy of each treatment arm was determined by Single Mean Test and the comparative efficacy was determined using Two Mean Test. All point estimates, such as mean and standard deviation and interval estimates such as 95% confidence intervals will be provided for the estimated parameters.
Detailed discussion of results and observations:
MTT assay was performed to test the cytotoxicity levels of the herbs used in the assay. A wide range of concentration series from 1ng to 1000µg/ml was found to be non-toxic to the cells. The viability was more than 90% even in the highest concentration tested. Fig 1 depicts the cell viability at the concentrations tested at 24 hrs post incubation with herbs.
Testosterone, the male sex hormone is endogenously expressed by the MA10 cells and were secreted into the cell supernatant at the concentration of 604pg/ml. i.e., 60.4pg testosterone was secreted by 8 X 104 MA10 cells as shown in Fig 2. This concentration increases to 781pg/ml when induced with Human chorionic gonadotrophin (HCG). This HCG induced 781pg/ml is further increased with 500µg/ml with P.ginseng (Herb A) to 822pg/ml. This P.ginseng induced testosterone synthesis is further increased in a dose dependent manner with increasing amounts of Herb B i.e., especially the maximum induction seen at 1:1 ratio of both the herbs.
This synergistic increase in testosterone production in leydig cancer cell line like MA10 very obviously gives strong evidence for the positive outcome of this herbal combination in improving male sexual health and general health.
NO-cGMP signaling is believed to play a major role in smooth muscle relaxation during sexual arousal and ejaculation, which is mostly attributed to the regulation of endothelial Nitric oxide production. Studies have indicated the post-translational regulation of nitric oxide synthase by testosterone. As we already are aware of the increase in testosterone levels in Leydig cell line (shown in Fig 3), we further wanted to check if there is a relevant increase in Nitric oxide levels in MA10 incubated with the herbs P.ginseng (A) and T.terrestris (B) aqueous extracts at ratios 1:1 to 1:50 (A:B) as depicted in Fig 3. There was a dose dependant increase in the Nitric oxide levels as evident from the increase in Nitrate/Nitrite levels (by products of Nitric oxide). Sodium nitroprusside were is a NO donor and is used as positive control. Endogenous Nitrate/Nitrite levels from 8 X 104 MA10 cells were 3.2 µM, which was improved to 12µM with 500µg/ml of herb B. This increase further increases in a dose dependant manner up to 15.5µM with both herbs A and B at equal ratios. This Synergistic effect of the herbal combination in in increasing the Nitric oxide levels in testicular leydig cell line is a strong evidence of the potential roles of this herbal combination in improvement of male sexual functions.
Double blinded clinical trial were conducted on healthy male volunteers for 2 months and have produced promising results on the efficacy of the drink in improvement of overall sexual functions. Two mean test was done to test whether Treatment Arm-I (male sexual wellness drink) is better than Treatment Arm-II (Placebo drink) in terms of whether the mean total Score of SIPFBQ is greater than 20. As shown in box and whisker plot in Fig 4, the average improvement from baseline to end of study was 45.19% in subjects who consumed Amore Male Sexual Wellness Drink in comparison with Placebo Drink. Most notably, the clinical trial indicated that 93% of the subjects who consumed Amore Male Sexual Wellness Drink showed =20% improvement in comparison with Placebo Drink.
As shown in Fig 5, there a very significant improvement of in MSQ score in the subjects who consumed the test drink for 2 months. The table in Figure 5 clearly shows the improvement in the score obtained for all the individual questions of the MSQ questionnaire. The mean percentage score for test Vs placebo arm for all the questions were strikingly significant numbers.
In the current invention, the scientific studies conducted both in cell based in vitro assays and clinical trials have strongly proven the efficacy of the unique herbal combination and the formulation in overall improvement of sexual and general wellbeing in men.
In an embodiment of the present invention, the formulation of Male sexual drink used in clinical trial is disclosed. The formulation comprising of Step 1: Disperse weighed quantity of xanthan gum in about 30 mL of warm, purified water to make a uniform dispersion of gum and allow to rest for about 15 minutes. Step 2: Dissolved the herbal extracts in about 20 mL of purified water under stirring. Step 3: To the above herbal extract solution add weighed quantities of sodium citrate, sucralose and sodium benzoate one after the other and mix for about 5 minutes. Add the solution of herbs slowly to xanthan gum under stirring. Step 4: To 30 mL of purified water add spray dried fruit powder and mix well without formation of lumps. Add fruit powder solution to the xanthan gum mixture and mix well. Step 5: To this mixture add colour and flavour one after the other and mix well. Step 6: To about 10 mL of water add citric acid, dissolve and add to the above mixture. Step 7: Transfer the mixture to a 100 mL measuring cylinder and make up the volume with purified water. Step 8 : Transfer again to the mixing vessel, mix for about 5 minutes. Step 9: Transfer the drink to the primary packing container, label and store in room temperature
Although this invention has been described by examples and with reference to possible embodiments thereof, it is to be understood that modifications or improvements may be made thereto without deviating from the scope of the invention. Further objectives, advantages, and features of the present invention will become apparent from the detailed description provided herein, in which various embodiments of the disclosed present invention are illustrated by way of example and appropriate reference to accompanying drawings. Those skilled in the art to which the present invention pertains may make modifications resulting in other embodiments employing principles of the present invention without deviating from its spirit or characteristics, particularly upon considering the foregoing teachings. Accordingly, the described embodiments are to be considered in all respects only as illustrative, and not restrictive, and the scope of the present invention is, therefore, indicated by the appended claims rather than by the foregoing description or drawings. Consequently, while the present invention has been described with reference to particular embodiments, modifications of structure, sequence, materials and the like apparent to those skilled in the art still fall within the scope of the invention as claimed by the applicant ,CLAIMS:1.An herbal sexual wellness formulation for males providing synergistic effects in vitro comprising of.
a.An extract of Ingredient A used in the concentration range of 100 – 700 µg/ml.
b.An extract of Ingredient B used in the range of 100 – 700 µg/ml.
Wherein, Ingredient A is aqueous extract from the root of the plant but not limited to fruit, leaf, stem, or inflorescence extract.
Wherein, Ingredient B is an aqueous extract from the whole dried fruit of the plant but not limited to stem, leaf, inflorescence, or root extract.
Wherein, the herbal extract is an aqueous extract but not limited to ethanol, methanol, acetone, ethyl acetate extracts and contains the bio-actives protodioscin, alkaloids, steroidal saponins, furostanol saponins, flavonoid glycosides, coumarins, alkaloids, and polyphenol compounds, and other bioactives.
2.The herbal hangover formulation as claimed in claim 1, wherein the herbal extract used as Ingredient B is obtained from Tribulus terrestris but not limited to T.terrestris orientalis, T.terrestris var robustus, T.terrestris var brachyceras, T.terrestris var inermis, T. terrestris var sericeus, T. terrestris var rajasthanensis, T.terrestris var macrocarpus, T.terrestris var desertorum, T.terrestris var biocornutus.

3.The herbal hangover formulation as claimed in claim 1, wherein the herbal extract used as Ingredient A is obtained from Panax ginseng but not limited to Panax quinquefolius var ginseng, P. notoginseng, Panax ginseng var. repens.
4.The herbal formulation as claimed in claim 1, has been evaluated invitro using cell lines, wherein the cell lines can be of mammalian, avian, Piscean, or arthropod origin, wherein, the cell type is Leydig cells but not limited to other carcinoma cell lines of testicular origin.
5.The herbal hangover formulation as claimed in claim 1, wherein at least one pharmaceutically accepted excipient is selected from, but not limited to, coloring agents , flavoring agents, acid stabilizers, preservatives, emulsifier/stabilizer/gelling agent/ thickener wherein, coloring agents is allura red, fruit powder as natural anti-oxidant but not limited to strawberry powder, , acidity regulator is sodium citrate but not limited to citric acid, acetic acid, malic acid, lactic acid, adipic acid and other salts, preservative is sodium benzoate but not limited to other similar preservatives, emulsifier/stabilizer/gelling agent/ thickener is xanthan gum but not limited to other similar agents..
6.The herbal hangover formulation as claimed in claim 1, wherein the formulation is an oral administration selected from, but not limited to, a pill, tablet, capsule, concentrated syrup, food additive, suspension, emulsion, powdered drink mix, carbonated/non-carbonated beverage and a chewable solid.
7.The herbal hangover formulation as claimed in claim 1, wherein the synergistic herbal formulation comprising of Panax ginseng and Tribulus terrestris along with other food grade ingredients improved the overall sexual function in healthy male volunteers
8.The clinical efficiency of the sexual wellness drink as mentioned in claim 7, wherein result of significant positive outcome was measured by MSQ, but not limited to other methods to measure the outcome of improvement of male sexual function
9.The clinical efficiency of the sexual wellness drink as mentioned in claim 7, wherein there was a sexual function improvement of 45.19% in test compared to 0% in placebo group.
The clinical efficiency of the sexual wellness drink as mentioned in claim 7, wherein 93% of the subjects showed =20% improvement from baseline to end of study. (Z = 1.68, p = 0.046 < 0.05) against none from the other arm that consumed placebo drink over the period of 2 months.