DESC:FIELD OF THE INVENTION:
The present invention relates to a topical formulation containing herbal composition for transdermal application. More particularly the present invention relates to an herbal composition for transdermal application is a cream formulation. The formulation is useful for a variety of cosmetic and/or therapeutic applications. The formulation is designed to deliver therapeutic levels of a drug which causes vasodilation. The present transdermal formulation is use for enhancing muscular definition, for the treatment of bruises on skin, in baldness, leg ulcers. More particularly the present topical formulation addresses the sexual dysfunction caused by impotency in males and vaginal dryness in females.

BACKGROUND OF THE INVENTION:
Approaches to improving blood flow to the skin have been many and consist of both systemic and topical approaches. Many beneficial effects can be experienced by a subject through an improvement in local blood flow, since impairment of local blood flow causes a variety of negative consequences. Among these are cold hands and feet, baldness, leg ulcers, certain forms of impotence, as well as a variety of other ailments.
While many of the currently available topical vasodilator formulations increase or enhance blood flow to some degree, there is nonetheless, a continued interest in identifying new formulations, which provide longer lasting vasodilator effects without undesirable side effects.
Accordingly, there is continued interest in the development new topical vasodilator formulation.
A bruise, also known as a contusion, is a common skin injury that results from the breakage of tiny blood vessels leaking under the skin. Blood from damaged blood vessels beneath the skin collects near the surface of the skin to appear as what we recognize as a black and blue mark. This mark is from skin discoloration by red blood cells and their contents. Generally, there are limited treatments for contusions to the skin. Vascular dilators may be administered in an amount sufficient to improve blood supply to the skin. Without wishing to be bound by theory, vascular dilators are also believed to strengthen blood vessels. Bruise creams generally are available to consumers, which typically include at least Arnica oil either alone or in combination with Witch Hazel or Menthol.
A need continues to exist for a treatment that reduces the duration of contusions of the skin. Human sexual responses in both men and women are due to a complex interaction of psychological, hormonal, and other physiological effects. One important aspect of the human sexual response common to both men and women is the erectile response that is itself due to interactions between the autonomic nervous system, the endocrine system, and the circulatory system.
Erectile response deficiencies are very common in men and have been referred to as impotence. Impotence refers to the inability of men to achieve or maintain penile erections sufficient for vaginal penetration and intercourse. Numerous approaches have been made as attempts to treat impotence.
The term sexual impotence, or inhibited sexual excitement, refers to the inability of a man to achieve a quality of erection sufficient to enable him to successfully experience coitus. The cause of this impotence may be either primary or secondary. In primary impotence, the man has never been able to achieve a satisfactory erection for coitus. In secondary impotence, the man has previously been potent and usually has been able to reach coitus but has subsequently developed his impotence.
However, with the advent of the so-called "sexual revolution" more men under this age group appear to be seeking treatment for this condition. The Kinsey statistics do reveal a gradual rise in the incidence of erectile dysfunction with age, particularly after 45, with a more rapid increase after 55. By age 75, nearly 55 percent of males reported problems, and by 80, 75 percent, However, research has shown (The Sexual Experience, (Sadock, Kaplan, and Freeman, eds.), Williams and Wilkins, Baltimore, 1976, Chapt. 15.4.) that aging is not an inevitable cause of impotence, even into the seventh and eighth decades of life.
Erectile dysfunction is almost always due to physical factors. Physical factors include systemic diseases (e.g. diabetes mellitus the most common!, syphilis, alcoholism, drug dependency, hypopituitarism, and hypothyroidism); local disorders (e.g. congenital abnormalities and inflammatory diseases of the genitalia); vascular disturbances such as aortic aneurysm and atherosclerosis (e.g. Leriche's syndrome): neurogenic disorders (e.g. multiple sclerosis, spinal cord lesions, pituitary microadenoma with hyperprolactinemia, and cardiovascular accident); drugs such as antihypertensives, sedatives, tranquilizers, and amphetamines; and surgical procedures such as sympathectomy. Prostatectomy and castration produce varying effects. Impotence is usually not induced by transurethral prostatectomy, whereas it almost always occurs after perineal prostatectomy. The Merck Manual, 16th edition, (Berkow, R., ed.), Merck Research Laboratories, Rahway, N.J., 1992, Chapt. 139.
Psychological factors, which include an abnormal fear of the vagina, sexual guilt, fear of intimacy, or depression, are the cause in about 20% of the cases of erectile dysfunction.
To date the pharmacological management of erectile dysfunction has been based mainly on the intracavernous injection of various smooth muscle relaxant drugs. Self-injection of smooth muscle relaxant drugs such as papaverine and/or prostaglandin E1 alone or in combination with phentolamine has been used successfully for a number of year in the treatment of impotence. However, numerous side effects have been reported and many patients have stopped using the self-injection procedure for various reasons mainly because of the lack of spontaneity and because of the unpleasant and sometimes pathological side effects caused by this technique. Consequently, a non-invasive therapeutic alternative seems attractive.
To this end, a number of investigators have begun to utilize preparations consisting of nitroglycerin in either ointment or patch form. In a study by Meyhoff, et al. (Meyhoff, H., Rosenkilde, P., and Bodker, Al., Brit. J. Urol., Vol. 68, pp. 89-90, (1992)) a nitroglycerin ointment was evaluated with good results and in a previous study by Claes and Baert (Owen, J. A., et al., J. Urol., Vol. 141, pp. 546548, (1989)) a nitroglycerin ointment was used with positive results. Both types of preparations suffer from one or more of the following disadvantages. The time necessary to achieve an erection varied widely with latent periods ranging from 1 to 2 hours. Consequently, any notion of spontaneity is invalidated. Both preparations primarily use lipophilic agents, such as petroleum jelly and/or lanolin as the matrix for the nitroglycerin. These agents lead to not only a very slow release of nitroglycerin into the penile tissue but also leave an oily residue on the penile shaft such that the nitroglycerin contained in this residue is easily transferred to the partner via the vaginal mucosa. A further drawback to the use of these preparations is the production of an intense burning sensation when the material is applied to the penis.
Concerning a different problem, no medications and/or viable treatment methods are currently available for the female who suffers from inhibited sexual excitement (i.e., anorgasmia or sexual arousal disorder), even though there are certain physiological and anatomical similarities between male and female external genitalia.
In the normal man or woman, a sequence of physiological sexual responses exists that has been described by Sadock et al., in Sadock, Kaplan, and Freeman, Eds., The Sexual Experience, Williams and Wilkins, Baltimore, (1976), Chapter 3. These levels of sexual arousal consist of four discrete phases, each accompanied by unique physiological changes. These phases can be understood physiologically as increasing levels of vasocongestion and myotonia (tumescence) and the subsequent rapid release of this vascular activity and muscle tone as a result of orgasm (detumescence).
If the clitoris does not become engorged with blood, for any number of reasons, orgasm is not attainable. Women who present this complaint are diagnosed as having a sexual arousal disorder. This condition is more precisely defined as the persistent or recurrent failure to attain or maintain the lubrication-swelling response of sexual excitement until completion of sexual activity. This inhibition occurs despite adequate sexual stimulation in focus, intensity, and duration. The disorder may be primary or, more frequently, secondary and restricted to the partner. Psychologically acquired factors cause most of the cases of secondary dysfunction; e.g., marital discord (about 80% of the cases), depression, and stressful life situations. Ignorance of genital anatomy and function is common, particularly of clitoral function and of effective arousal patterns and techniques. Association of sex with sinfulness, and sexual pleasure with guilt, may be lifelong. Fear of intimacy may also play a part.
The physical causes include localized disease (e.g., endometriosis, cystitis, vaginitis); systemic diseases (e.g., hypothyroidism, diabetes mellitus--though its impact is greater on men); peripheral of CNS disorders (e.g., multiple sclerosis); muscular disorders (e.g., muscular dystrophy); drugs (e.g. oral contraceptives, antihypertensives, tranquilizers have variable effects); and ablative surgery (e.g., hysterectomy, mastectomy, which may have a negative impact on the woman's sexual self-image).
Besides those women in the general population, recent publications have emphasized these problems in older female populations (Osburn, M., Brit Med. J., Vol. 296, pp. 959-962, (1988)), in post-menopausal women, and in those who have undergone a hysterectomy (The American College of Obstetricians, Gynecologists Office Practice and Practice Management: Sexuality and Sexual Dysfunction, American College of Obstetricians and Gynecologists, Chapter 10 (1986)). In addition, those women afflicted with Type II diabetes appear to suffer from an increased frequency of sexual problems, including anorgasmia (Schreiner, et al., "Diagnosing and Treating the Sexual Problems of Diabetic Women," in Clinical Diabetes, Vol 6, pp. 121-136 (1988)).
Although there are many reasons given for the inability of a woman to reach orgasm, one aspect of the present invention is directed toward the first phase of the sexual response cycle, the excitement phase. Since the female clitoris and its associated structures are both anatomically and embryologically similar to the male penis, it is medically correct to assume that the clitoris should (and does) respond in kind during the excitement phase of the response cycle.
Both the clitoris and the penis have corpora cavernosa, an anatomical shaft, and both are erectile. Consequently, since a satisfactory erection can be produced in impotent males by the introduction of vasoactive agents into the male members, either by injection or by transdermal methods, it follows that the same procedure(s) will be equally valid in treating women who complain of inhibited sexual excitement. However, the introduction of vasodilators or other vasoactive agents by hypodermic injection would be both impractical and dangerous.
Further, since the clitoris is much smaller than its male counterpart, has a much thinner striatum cornuium, and is more sensitive to tactile stimulation, any topical transdermal preparations containing a vasodilator must address these characteristics.
There continues to be a need in the art for effective means to modulate the sexual response of humans, particularly to enhance the erectile abilities of men suffering from impotence. Such means are ideally convenient and simple to use, do not need to be administered at a constant dose or administered multiple times to achieve the desired results, are not invasive, and are not required and will allow a rapid and predictable ability for the onset of erectile function in response to and in response to normal sexual stimulation.

SUMMARY OF THE INVENTION:

The principal aspect of the present invention to provide a herbal composition comprising extracts or fractions derived from Salvia tomentosa (Balsamic Sage), and/or Thymus vulgaris (Thyme), and/or Marigold and topical formulation thereof, for vasodilatation.
Another objective of the present invention is to provide a topical formulation containing herbal composition comprises Luteolin as an active ingredient in present formulation for transdermal application.
In still another embodiment of the present invention is an herbal formulation, wherein the said composition is in form of cream.
Yet another objective of the present formulation is directed to transdermal formulations for increasing or enhancing blood flow in a subject.
Another aspect of the present invention discloses aforesaid formulation for herbal composition containing Humectant, Emulsifier – Lubricant, Penetration Enhancer, Fragrance and Distilled Water.
Another object of the present invention is to disclose composition of cream formulation for herbal composition wherein Humectant is glycerin or sorbitol or mixture thereof.
Another object of the present invention is to disclose composition of cream formulation for herbal composition wherein Emulsifier – Lubricant is stearic acid or bees wax or carnauba wax or mixture thereof.
Another object of the present invention is to disclose composition of cream formulation for herbal composition wherein penetration enhancer is Jojoba oil or ethanol or mixture thereof.
Another object of the present invention is to disclose composition of cream formulation for herbal composition wherein emulsifying stabilizer is stearic acid or emulsifying wax or mixture thereof.
In another aspect of the present invention it contains present formulation vitamins molecule can be selected from various forms of tocopherol, shows activity of vitamin E.
In yet another aspect of the present invention relates to vasodilator formulations, for increasing, enhancing and/or stimulating blood flow to assist in the therapeutic treatment of diseases and/or conditions such as for example erectile dysfunction, bruise, aging, baldness, peripheral neuropathy diabetes and in such as for example the enhancement of muscle and vein definition in body builders, physique models, models, and/or aesthetically conscious individuals, thus enhancing the muscle definition.
In one embodiment, the disease, disorder or condition associated with a decreased or impaired blood flow and/or vasoconstriction is selected from the following non-limiting examples, including erectile dysfunction, aging, baldness, peripheral neuropathy, microangiopathy, female sexual dysfunction, male sexual dysfunction, diabetes, aging, restless leg syndrome, raynaud's phenomenon, Buerger's Disease, chilblains, numbness and tingling of extremities, varicose veins, haemorrhoids, hypothyroidism, immobility, cellulite, accumulation of subcutaneous adipose tissue, cosmetic applications such as poor quality hair, nail and skin, lymphedema, swelling of the hands and feet oedema, deep vein thrombosis, ischemia, chronic venous insufficiency, gangrene, vasoconstriction, thrombosis, embolism, paraesthesia, poikilothermia, cellulitis, tissue necrosis, ischaemic neuropathy, leg cramps either idiopathic, or related to either pregnancy, renal dialysis, or peripheral vascular disease (both venous and arterial), or to revitalize muscle, or to improve muscle strength and recovery after vigorous exercise and intense sport, or to improve muscle strength and performance during vigorous exercise and intense sport.

The present invention relates to the formation of cream formulation and methods for using said cream formulation more particularly in treating sexual dysfunction, including impotency or erectile dysfunction in males, and sexual dysfunction in females caused by vaginal dryness.

In one embodiment, the subject is a mammal preferably the subject is a human. Alternatively, the subject is a non-human mammal, non-limiting examples of which include a. horse, dog, cat, rabbit and the like.

DETAILED DESCRIPTION OF THE INVENTION:
The present invention in detailed provides a topical formulation containing herbal composition comprising Luteolin; acting as an active ingredient in present formulation for transdermal application.
In still another embodiment of the present invention is an herbal formulation, wherein the said composition is in form of cream.
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. However, any skilled person will appreciate the extent to which such embodiments could be extrapolated in practice.
The present formulation is useful for variety of cosmetic and/or therapeutic applications. Further the present formulation is designed to deliver therapeutic levels of a drug which causes vasodilation. The present transdermal formulation is use for enhancing muscular definition, for the treatment of bruises on skin, in baldness, leg ulcers. More particularly the formulation addresses the sexual dysfunction caused by impotency in males and vaginal dryness in females.

The present invention of cream formulation discloses herbal composition vasodilation effect comprising extracts or fractions derived from Salvia tomentosa (Balsamic Sage), and/or Thymus vulgaris (Thyme), and/or Marigold. The present formulation of cream is for increasing blood flow through vasodilation for enhancing muscular definition, treat bruises on skin and more particularly sexual dysfunction caused by impotency in males and vaginal dryness in females.
The present cream formulation contains active ingredient as a Luteolin; and excipients to form cream base along with some acceptable additives and fragrance to add refreshing effect in present cream formulation.
In yet another embodiment, herbal composition wherein the plant extracts are obtained from plant parts selected from leaf, seeds, floral parts rhizome and aerial parts.
Creams are semi-solid emulsions of oil and water. They are divided into two types: oil-in-water (O/W) creams which are composed of small droplets of oil dispersed in a continuous water phase, and water-in-oil (W/O) creams which are composed of small droplets of water dispersed in a continuous oily phase. Oil-in-water creams are more comfortable and cosmetically acceptable as they are less greasy and more easily washed off using water. Water-in-oil creams are more difficult to handle but many drugs which are incorporated into creams are hydrophobic and will be released more readily from a water-in-oil cream than an oil-in-water cream. Water-in-oil creams are also more moisturising as they provide an oily barrier which reduces water loss from the stratum corneum, the outermost layer of the skin.
Transdermal delivery systems (TDS) are self-contained, discrete dosage forms that, when applied to intact skin, are designed to deliver the drug(s) through the skin to the systemic circulation. Systems typically comprise an outer covering (barrier), a drug reservoir that may have a drug release–controlling membrane, a contact adhesive applied to some or all parts of the system and the system/skin interface, and a protective liner that is removed before the patient applies the system. The dose of these systems is defined in terms of the release rate of the drug(s) from the system and surface area of the patch and is expressed as mass per unit time for a given surface area. With these drug products, the skin typically is the rate-controlling membrane for the drug input into the body. The total duration of drug release from the system and system surface area may also be stated. TDS work by diffusion: The drug diffuses from the drug reservoir, directly or through the rate-controlling membrane and/or contact adhesive if present, and then through the skin into the general circulation. Typically, modified-release systems are designed to provide drug delivery at a constant rate so that a true steady-state blood concentration is achieved and maintained until the system is removed. Following removal of the system, blood concentration declines at a rate consistent with the pharmacokinetics of the drug.

LUTEOLIN
Natural resource:
Luteolin is a flavone, a type of flavonoid, with a yellow crystalline appearance. Luteolin is most often found in leaves, but it is also seen in rinds, barks, clover blossom, and ragweed pollen. It has also been isolated from the aromatic flowering plant, Salvia tomentosa in the mint family, Lamiaceae.
Dietary sources include celery, broccoli, green pepper, parsley, thyme, dandelion, perilla, chamomile tea, carrots, olive oil, peppermint, rosemary, navel oranges, and oregano. It can also be found in the seeds of the palm Aiphanes aculeata.
In the present formulation comprising Luteolin as an active ingredient that has been extracted or fraction derived from Salvia tomentosa (Balsamic Sage), and/or Thymus vulgaris (Thyme), and/or Marigold.
THYMUS VULGARIS (Thyme):
Thymus Vulgaris (common thyme, German thyme, garden thyme, or just thyme) is a species of flowering plant in the mint family Lamiaceae, native to southern Europe from the western Mediterranean to southern Italy. Growing to 15–30 cm (6–12 in) tall by 40 cm (16 in) wide, it is a bushy, woody-based evergreen subshrub with small, highly aromatic, grey-green leaves and clusters of purple or pink flowers in early summer.
It is useful in the garden as groundcover, where it can be short-lived, but is easily propagated from cuttings. It is also the main source of thyme as an ingredient in cooking and as an herbal medicine. It is slightly spicier than oregano and sweeter than sage.
Botanical Classification
Kingdom: Plantae
Sub kingdom Tracheophytes
Division Angiosperms
Class Eudicots / Magnoliopsida
Sub class Asteridae
Order: Lamiales
Family: Lamiaceae
Genus: Thymus
Species: T. vulgaris

SALVIA TOMENTOSA (Balsamic Sage)
Salvia is the largest genus of plants in the mint family, Lamiaceae, with nearly 1000 species of shrubs, herbaceous perennials, and annuals. Within the Lamiaceae, Salvia is part of the tribe Mentheae within the subfamily Nepetoideae. One of several genera commonly referred to as sage, it includes the widely produced herb used in cooking, Salvia officinalis (common sage, or just "sage").
The genus is distributed throughout the Old World and the Americas, with three distinct regions of diversity: Central and South America (approx. 500 species); Central Asia and Mediterranean (250 species); Eastern Asia (90 species).
Many interspecific hybrids occur naturally, with a relatively high degree of crossability, but some such as S. officinalis × S. lavandulifolia and S. fruticosa × S. tomentosa have been intentional. A natural hybrid, S. longispicata × S. farinacea has given rise to a series of popular ornamentals such as S. 'Indigo Spires' and S. 'Mystic Spires Blue'

Botanical Classification
Kingdom: Plantae
Sub kingdom Tracheophytes
Division Angiosperms
Class Eudicots / Magnoliopsida
Sub class Asteridae
Order: Lamiales
Family: Lamiaceae
Sub Family Nepetoideae
Genus: Salvia
Species: S. tomentosa

CALENDULA OFFICINALIS (The marigol flowers )
The results of the HPLC analysis indicated that marigold flowers extract contain nine different active compounds, including Vitexin 11.40%, Rutin 12.29%, Quercetin-3-3galactosid 12.64%, Luteolin-7-glucose9.27%, Quercetin-3- glucoside 7.38%, Quercitrin 9.83%, Myricetin 10%, Luteolin 10.72%, Apigenin7.08% and kampferol 9.37 %. The results of a clinical study showed the effect of marigold flowers extract cream as an antioxidant which protected the skin in particular from oxidative damage after sunburn and reduced the symptoms of skin aging. This effect was evident in both concentrations 10% and 15% when compared with control. Control response rate, 10% of marigold flowers cream and 15% of flowers extract cream were reached to 76% , 85% and 92% within two weeks of treatment respectively. The Conclusion of this study showed the importance of marigold flowers extract as a source of bioactive compounds such as rutin and quercetin derivatives, vitexin, luteolin, apigenin and kampferol which act as an antioxidant to restore skin health and aging resistance. This study approved that marigold flowers extract characteristics can make it the main ingredients in the preparation of topical agents for the treatment of various skin diseases.

Botanical Classification
Kingdom: Plantae
Sub kingdom Tracheophytes
Division Angiosperms
Class Eudicots / Magnoliopsida
Sub class Asteridae
Order: Asterales
Family: Asteraceae
Genus: Calendula
Species: Calendula officinalis
Chemical nature of Luteolin:
Luteolin is a naturally occurring flavonoid. The flavonoids are polyphenolic compounds found as integral components of the human diet. They are universally present as constituents of flowering plants, particularly of food plants. The present formulation contains Luteolin with formula I as an active ingredient for vasodilatation. The flavonoids are phenyl substituted chromones (benzopyran derivatives) consisting of a 15-carbon basic skeleton (C6-C3-C6), composed of a chroman (C6-C3) nucleus (the benzo ring A and the heterocyclic ring C), also shared by the tocopherols, with a phenyl (the aromatic ring B) substitution usually at the 2-position. Different substitutions can typically occur in the rings, A and B.

Molecular Formula: C15H10O6

SYNONYM
• 3', 4’, 5, 7-Tetrahydroxyflavone
• Digitoflavone
• Luteolol
• Flacitran
• 2-(3,4-Dihydroxyphenyl)-5,7-dihydroxy-4H-chromen-4-one

IUPAC NAME:
2-(3, 4-dihydroxyphenyl)-5, 7-dihydroxychromen-4-one
Pharmacology of Luteolin:
Luteolin have numerous beneficial medicinal properties such as anti-cancer, anti-oxidant, anti-inflammatory, and anti-allergic actions, anti-lipidermic action and increasing eNOS (endothelial Nitric Oxide Synthase) gene expression. Luteolin elicited relaxation of potassium contracted aortic rings which results in vasorelaxation effect attenuated by the eNOS inhibitor, N-nitro-L-arginine methyl ester, suggesting that this Luteolin action is at least partially mediated by activating eNOS activity and increased eNOS phosphorylation and NO production. Luteolin can directly act on vascular ECs (Endothelial Cells), leading to rapid eNOS activation and NO production, thereby causing relaxation of vascular tension which increases the blood flow at the muscle site and rate of Muscle pumping.
In present formulation Luteolin enhances exercise sprint performance, likely by improving brain oxygenation and allowing a higher muscle extraction of oxygen. It improves the muscle mass by improving vasodilation by acting as a nitric oxide donor to blood vessels. It is most efficient flavonoid for the muscle mass building and anti-stress agent.
Several plants and spices containing flavonoid derivatives have found application as disease preventive and therapeutic agents in traditional medicine in Asia for thousands of years. The selection of a particular food plant, plant tissue or herb for its potential health benefits appears to mirror its flavonoid composition. The much lower risk of colon, prostate and breast cancers in Asians, who consume more vegetables, fruits and tea than populations in the Western hemisphere do, raises the question of whether flavonoid components mediate the protective effects of diets rich in these foodstuffs by acting as natural chemopreventive and anticancer agents. An impressive body of information exists on the antitumoral action of plant flavonoids. In vitro work has concentrated on the direct and indirect actions of flavonoids on tumor cells, and has found a variety of anticancer effects such as cell growth and kinase activity inhibition, apoptosis induction, suppression of the secretion of matrix metallo-proteinases and of tumor invasive behavior.
Luteolin also acts on smooth muscle as relaxation of the smooth muscles of the body Luteolin is considered to be an endocrine disruptor with potent estrogen agonist activity and potent progesterone antagonist activity. Luteolin has effects on smooth muscle. Luteolin relaxed smooth muscle as it inhibits both phosphodiesterase and reduced intracellular Ca2+ entry and intracellular Ca2+ release. Luteolin also caused vasorelaxation of smooth muscle by inhibiting intracellular Ca2+ release, inhibition of sarcolemmal Ca2+ channels, and activation of K+ channels. Thus it acts on smooth muscle and found to be potent smooth muscle relaxant.
Furthermore, some studies have reported the impairment of in vivo angiogenesis by dietary flavonoids. Experimental animal studies indicate that certain dietary flavonoids possess antitumoral activity. The hydroxylation pattern of the B ring of the flavones and flavonols, such as luteolin seems to critically influence their activities, especially the inhibition of protein kinase activity and antiproliferation. The different mechanisms underlying the potential anticancer action of plant flavonoids await further elucidation. Certain dietary flavonols and flavones targeting cell surface signal transduction enzymes, such as protein tyrosine and focal adhesion kinases, and the processes of angiogenesis appear to be promising candidates as anticancer agents. Further in vivo studies of these bioactive constituents is deemed necessary in order to develop flavonoid-based anticancer strategies. In view of the increasing interest in the association between dietary flavonoids and cancer initiation and progression, this important field is likely to witness expanded effort and to attract and stimulate further vigorous investigations.
Luteolin is a naturally-occurring flavonoid, with potential anti-oxidant, anti-inflammatory, apoptosis-inducing and chemopreventive activities. Upon administration, Luteolin scavenges free radicals, protects cells from reactive oxygen species (ROS)-induced damage and induces direct cell cycle arrest and apoptosis in tumor cells. This inhibits tumor cell proliferation and suppresses metastasis.

CREAM FORMULATION:
Creams are multiphase preparations consisting of a lipophilic phase (oil phase) and an aqueous phase.
Lipophilic creams: Lipophilic creams have as the continuous phase (larger component) the lipophilic phase. They usually contain water-in-oil emulsifying agents such as lanolin, sorbitan esters and monoglycerides.
Hydrophilic creams: Hydrophilic creams have as the continuous phase (larger component) the aqueous phase. They contain oil-in-water emulsifying agents such as sodium or trolamine soaps, sulfated fatty alcohols, polysorbates and polyoxyl fatty acid and fatty alcohol esters combined, if necessary, with water-in-oil emulsifying agents.
Creams are semisolid dosage forms containing one or more drug substances dissolved or dispersed in a suitable base. This term has traditionally been applied to semisolids that possess a relatively fluid consistency formulated as either water-in-oil or oil in water emulsions. However, more recently the term has been restricted to products consisting of oil-in-water emulsions or aqueous microcrystalline dispersions of long-chain fatty acids or alcohols that are water washable and more cosmetically and aesthetically acceptable.
The choice of cream base depends on the drug, base compatibility, absorption: skin properties, blood flow and type of wound. The main consideration is the consistency of the expected dosage, the properties of the substance used and the basic requirements include: non-irritant, easy to clean, not left on the skin, stable not dependent on pH, homogenous with various drugs.
The cream base consists of a base type W / O and O / W emulsion:
- W / O type emulsion base. Example: lanolin, cold cream. The properties of the W / O type emulsion base are emollient, occlusive, contain water, some absorb added water, and oily.
- O / W type emulsion base. Example: hydrophilic ointment. The properties of the O / W type emulsion base are easily washed with water, not greasy, can be diluted with water, not occlusive.
During use, a continuous phase will evaporate, and increase the concentration of water soluble substances in the attached layer. To prevent the deposition of drugs, and to increase absorption through the skin, added substances that are mixed with water but do not evaporate, namely humectant. A better formulation is a cream that can deposit fat and other moisturizing compounds to help hydrate the skin.
The emulsion base consists of three components, namely the oil phase, water phase and emulsifier.
The oil phase:
Oils make up, anywhere between 11 and 24% of the bulk of a cream. The heavier purely medicinal creams contain a higher proportion of oil but water is still their major ingredient. Oil-soluble herbal ingredients like resins dissolve and become incorporated into this phase which gives creams a richer and heavier feel. Many oils are susceptible to oxidation or rancidification over a period of time. This process is hindered by the addition of antioxidants like vitamin E, to all our creams and lotions. The oil phase is usually formed from petrolatum or liquid petrolatum with one or more high molecular weight alcohols such as cetyl or stearyl alcohol. Stearil alcohol and petrolatum form the oil phase which has the purpose of softening and making the skin comfortable. Stearyl alcohol also acts as an emulsifying adjuvant.
In the present invention ingredients stearic acid, jojoba oil, coco butter, olive oil comprises the oil phase for the cream formulation.
The aqueous (water) phase:
Water constitutes the major ingredient (61-77%) of most creams. The lighter and more cosmetic-type creams contain more water and less oil. This phase contains the water-soluble herbal ingredients of a cream. In our creams, the water phase is never tap or deionised water as is the case with most creams, except in our base cream. We always incorporate beneficial high quality active ingredients into the water phase in the form of distilled aromatic waters, strong infusions and decoctions and cold percolates of organic herbs.
The water phase contains preservatives, emulsifiers or parts of emulsifiers and humectants. Humectants are usually in the form of glycerine, propylene glycol or polyethylene glycol. The water phase can also contain water-soluble components from the emulsion system, along with other additives such as stabilizers, antioxidants, buffers etc.
In the present invention ingredients Glycerine, aloe vera extract or gel, demineralised water, sorbitol, and perfume comprises the aqueous phase for the cream formulation.
Emulsifier:
The ideal emulsifiers must stable, inert, free of toxic and irritant ingredients, odourless, tasteless and colourless, produces a stable emulsion of the desired type. The emulsifying agent consists of anionic emulsifiers, cationic and non-ionic emulsifiers. The main consideration in choosing an emulsifiers is the comparison of hydrophilic and lipophilic groups. Hydrophilic Lipophilic Balance (HLB) is a measure of the balance of the state of lipophilic and hydrophilic which is a characteristic of the surfactant group emulsifiers.
The most important step in producing an emulsion is the selection of a suitable emulsifier. The most commonly used emulsifiers are anionic and nonionic systems, or combinations of the two. The primary selection is based on the emulsion type (oil/water or water/oil) and the emulsifier must be compatible with the oil phase, water phase and the actives used in the formulation. The primary emulsifier is the main determinant factor of the emulsion type, while the secondary emulsifier serves to strengthen the interphase thereby improving its stability. The use of sodium soaps is often rejected, as they are too alkaline. When neutralized they produce a hard soap with limited solubility. Potassium Stearate is one of the most widely used emulsifiers as they are cheap and readily available. But it is irritating to the skin the skin and has a tendency to gel with time. A number of other factors like mildness on skin, color, odor, emollient effect and solubility in both the phases would also be considered while selecting the emulsifier system. The emulsifiers to be incorporated in the herbal compositions are selected from stearic acid, sodium stearate, potassium stearate, PEG 400 stearate, Glyceryl monostearate, Arlacel 165, bees wax, carnauba wax, emulsifying wax or mixture thereof.
Stearic acid:
Stearic Acid may be used to form the base of other ingredients that are intended to be incorporated into formulations as lubricants, emollients, and emulsifiers. In emulsions, Stearic Acid is an effective stabilizer, thickener, and softener that contributes a cooling sensation on the skin. It is also known to contribute a pearly finish to lubricants.
Bees wax or carnauba wax:
Used to thicken formulations, with emollient and protective qualities, waxes can provide stability to cream and topical applications and boost their viscosity and consistency.
Beeswax is naturally produced in the bee hive or honeycomb. Beeswaxes are BP/USP certified and come in white or yellow colour. Carnauba wax is obtained from the leaves of the Carnauba palm of Brazil. It is used in many polishes and personal care products as it imparts excellent shine and hardness.
Emulsifying wax:
Emulsifying Wax is the chemical uses to bind oil and water together to form a smooth lotion, cream and emulsion.
Emulsifying wax is a cosmetic emulsifying ingredient. The ingredient name is often followed by the initials NF, indicating that it conforms to the specifications of the National Formulary.
Emulsifying wax is created when a wax material (either a vegetable wax of some kind or a petroleum-based wax) is treated with a detergent (typically sodium dodecyl sulfate or polysorbates) to cause it to make oil and water bind together into a smooth emulsion. It is a white waxy solid with a low fatty alcohol odor.
The ingredients for emulsifying wax NF are cetearyl alcohol and a polyoxyethylene derivative of a fatty acid ester of sorbitan (a polysorbate)
In a cream product, if the emulsifying wax used meets the standards for the National Formulary, it may be listed in the ingredient declaration by the term "emulsifying wax NF". Otherwise, the emulsifier is considered a blended ingredient and the individual components must be listed individually in the ingredient declaration, placed appropriately in descending order of predominance in the whole. In present cream formulation also it acts as an emulsifier and stabilizer.
Humectant:
Humectants (or moisturizers) are important cream ingredients allowing to prevent loss of moisture thereby retaining the skin's natural moisture. Some compounds also have the ability to actively attract moisture. Humectants are key ingredients in most skin care products but are also often used in hair care products to volumize the hair by attracting moisture which expands the hair shaft. There is a large variety of very different compounds providing moisturizing effects including proteins, acids, polysaccharides, and various small molecules.
In present invention humectant is selected from sorbitol or urea or aloe vera or urea or glycolsor essential oils like eucalyptus oil, jojoba oil, peppermint oil, fennel oil, almond oil or mixture thereof.
Sorbitol:
Natural polyhydric alcohol derived from a sugar solution (dextrose) from wheat (gluten-free), consists of the alcohols D-glucitol & D-mannitol. White, crystalline powder, slightly sweet odor. Soluble in water or alcohols. Effective moisturizing properties (avoids moisture loss). It acts as a stabilizes gels and provides good clarity, has skin softening and conditioning effects. In present formulation it support as a moisturizer and preservative.
Sorbitol is also used as a humectant moisturizer in as creams formulation. A humectant is a hygroscopic substance that often has a molecular structure with several hydrophilic groups. This structure allows humectants to attract and retain the moisture in the air nearby via absorption, drawing the water vapor into or beneath the surface.
Glycerin:
In present cream formulation glycerin acts as a neutralizing agent and as a moisturizer. Glycerin is a humectant, a type of moisturizing agent that pulls water into the outer layer of your skin from deeper levels of your skin and the air. In skin care products, glycerin is commonly used with occlusive, another type of moisturizing agent, to trap the moisture that it draws into the skin.
Permeation Enhancer:
The human skin serves as an impediment, a thermo regulator and prevents excessive loss of water from the internal organs. Various ways of transferring the drugs have been developed by modifying the barrier properties of the skin. Enhancement in skin penetration by hydration of the stratum corneum, or by use of chemical enhancers acting on the lipids and keratinized structures in the stratum corneum, partitioning and solubility effects is a promising tool in potential clinical applications. Penetration enhancement is a new emerging technology which has the potential to increase the number of drugs taken trans-dermally. Also the drugs with short biological half-life could be easily administered. Among many advantages over other routes the three crucial ones are avoiding metabolism in liver, minimal negative effects and increased bioavailability. Also, the stratum corneum prevents the loss of physiologically essential substances and as a result provides penetration resistance by acting as a protective barrier. This is the rate limiting step in the absorption of the drug percutaneously.
Permeation enhancers are those substances which promote the absorption of drug through the skin temporarily by transiently enhancing the skin permeability. They are employed to transfer the delivery of drugs which are ionizable and impermeable; to maintain drug levels in blood, to provide higher dose of less potentially active drugs, to deliver high molecular weight hormones and peptides and to lessen the lag time of transdermal drug delivery system.
For the present invention the permeation enhancers are selected from Ethanol, dimethyl sulfoxide, dimethyl isosorbide, Isopropyl myristate and propylene glycol, eucalyptus oil, jojoba oil, peppermint oil, fennel oil, almond oil or mixture thereof.
Preservatives:
Preservatives are used to retard microbial (bacterial and fungal) contamination of cosmetics. The high water content of many cosmetics makes them suitable places for microbial growth. Combinations of preservatives are frequently used. The preservatives would include a combination of any 2-5 preservatives from the list - methyl paraben, propyl paraben, butyl paraben, Quaternium-8, -14 or -15, imidazolidinyl urea, diazolidinyl urea, phenoxy ethanol, citric acid and ethanol.
For the present invention preservative for cream formulation is selected from methyl paraben, propyl paraben, butyl paraben,phenoxy ethanol, citric acid,Sorbic Acid/Potassium sorbate, Levulinic Acid, Anisic Acid and ethanol.
Ethanol:
Ethanol in cream formulation or topical applications acts as a skin penetration enhancer and facilitate the transdermal absorption of active ingredients also. In the present cream formulation it also acts as a solvent vehicle. In cream formulation it acts as a preservative for increasing the shelf life of all the ingredients in the composition.
Jojoba oil:
In the present formulation it acts as moisturizer and skin penetration for getting better result as a transdermal formulation. In most of the transdermal application like cream, lotion, ointments it acts a preservative also.
Fragrance:
The fragrance(s) that may be included in the composition are not relevant to the inventive concepts disclosed herein, and those skilled in the art are familiar with the wide range of fragrances available. Therefore, any suitable fragrance or combination of fragrance natural and/or artificial, is within the contemplated scope of the present disclosure.
It is understood, therefore, that this invention is not limited to the particular embodiments or examples disclosed, but is intended to cover modifications within the objectives and scope of the present invention as defined in the specification. The presented examples illustrate the invention, but they should not be considered to limit the scope of the invention in any way.
The following example describe the invention and are in no way limiting the scope of the invention.

Formulation of cream comprising herbal composition for addressing sexual dysfunction:

Sr. No. INGREDIENTS CONCENTRATION
1 Luteolin (1 to 98%) – from

Salvia tormentosa extract
or
Apium graveolens(Celery) extract
or Capsicum annum (green pepper)extract
or Thymus vulgaris(Thyme) extract
or Matricaca Chamomilla extract
or Chrysanthemum morifolium extract
or Loricena japonica extract
1 to 75 %
2 Lemongrass oil 0.1 to 20%
3 Orange Oil 3 to 15 %
4 Cypress Oil 0.5 to 15%
5 Ginger Oil 0.1 to 8%
6 Eucalyptus Oil 1 to 25%
7 Tocopherol 0.1 to 5%
8 Stearic acid 2 to 20%
9 Emulsifying Wax 2 to 18%
10 Carnauba Wax 2 to 12%
11 Jojoba Oil 2 to 18%
12 Aloe-vera L/E 2.5 to 10%
13 Ethanol 5 to 15%
14 Sorbitol 2 to 8%
15 Glycerine 5 to 15%
16 Na-Methyl Paraben 0.1 to .15%
17 Phenoxy-ethanol 0.1 to 0.3%
18 Distilled Water q.s

METHOD OF PREPARATION:
1. In one container, Dissolve Aloe Vera Extract/Gel and Luteolin in same with Ethanol and half quantity of water.
2. In another Container, Dissolve selected Emulsifier with continuous temperature and stir until no lumps observed.
3. Mix Step 1 and Step 2 with addition of stearic acid and Tocopherol (Vitamin-E) until it becomes liquid with smooth texture.
4. Mix all other essential oils, carrier oil and preservative in above step 3.
5. Dissolve and make proper liquid phase in emulsifier tank by mixing water phase and remaining quantity of water.
6. Mix Step 4 in Step 5.
7. Continue to mix until it become proper blend and consistent Cream.

EXAMPLE: 1

Formulation of cream comprising herbal composition for addressing sexual dysfunction:

Sr. No. INGREDIENTS CONCENTRATION
1 Luteolin (1 to 98%) – from
Salvia tormentosa extract
or
Apium graveolens(Celery) extract
or Capsicum annum (green pepper)extract
or Thymus vulgaris(Thyme) extract
or Matricaca Chamomilla extract
or Chrysanthemum morifolium extract
or Loricena japonica extract 2%
2 Lemongrass oil - 3%
3 Orange Oil - 3%
4 Cypress Oil - 1.5%
5 Ginger Oil - 0.5%
6 Eucalyptus Oil - 19%
7 Tocopherol - 0.450%
8 Stearic acid - 18%
9 Emulsifying Wax - 7%
10 Carnauba Wax - 7%
11 Jojoba Oil - 9%
12 Aloe-vera L/E - 2%
13 Ethanol - 2.5%
14 Sorbitol - 3%
15 Glycerine - 6%
16 Na-Methyl Paraben - Quantity S.
17 Phenoxy-ethanol - Quantity S.
18 Distilled Water - Quantity S.

METHOD OF PREPARATION:
1. In one container, Dissolve Aloe Vera Extract/Gel and Luteolin in same with Ethanol and half quantity of water.
2. In another Container, Dissolve selected Emulsifier with continuous temperature and stir until no lumps observed.
3. Mix Step 1 and Step 2 with addition of stearic acid and Tocopherol (Vitamin-E) until it becomes liquid with smooth texture.
4. Mix all other essential oils, carrier oil and preservative in above step 3.
5. Dissolve and make proper liquid phase in emulsifier tank by mixing water phase and remaining quantity of water.
6. Mix Step 4 in Step 5.
7. Continue to mix until it become proper blend and consistent Cream.

EXAMPLE: 2

Formulation of cream comprising herbal composition for addressing sexual dysfunction:

Sr. No. INGREDIENTS CONCENTRATION
1 Luteolin (1 to 98%) – from
Salvia tormentosa extract
or
Apium graveolens(Celery) extract
or Capsicum annum (green pepper)extract
or Thymus vulgaris(Thyme) extract
or Matricaca Chamomilla extract
or Chrysanthemum morifolium extract
or Loricena japonica extract - 10%
2 Lemongrass oil 4%
3 Orange Oil 2%
4 Cypress Oil 2%
5 Ginger Oil 1%
6 Eucalyptus Oil 12%
7 Tocopherol 0.40%
8 Stearic acid 16%
9 Emulsifying Wax 5%
10 Carnauba Wax 4%
11 Jojoba Oil 8%
12 Aloe-vera L/E 2.5%
13 Ethanol 3%
14 Sorbitol 3%
15 Glycerine 6%
16 Na-Methyl Paraben Quantity S.
17 Phenoxy-ethanol Quantity S.
18 Distilled Water Quantity S.

METHOD OF PREPARATION:
1. In one container, Dissolve Aloe Vera Extract/Gel and Luteolin in same with Ethanol and half quantity of water.
2. In another Container, Dissolve selected Emulsifier with continuous temperature and stir until no lumps observed.
3. Mix Step 1 and Step 2 with addition of stearic acid and Tocopherol (Vitamin-E) until it becomes liquid with smooth texture.
4. Mix all other essential oils, carrier oil and preservative in above step 3.
5. Dissolve and make proper liquid phase in emulsifier tank by mixing water phase and remaining quantity of water.
6. Mix Step 4 in Step 5.
7. Continue to mix until it become proper blend and consistent Cream.

EXAMPLE: 3

Formulation of cream comprising herbal composition for addressing sexual dysfunction:

Sr. No. INGREDIENTS CONCENTRATION
1 Luteolin (1 to 98%) – from
Salvia tormentosa extract
or
Apium graveolens(Celery) extract
or Capsicum annum (green pepper)extract
or Thymus vulgaris(Thyme) extract
or Matricaca Chamomilla extract
or Chrysanthemum morifolium extract
or Loricena japonica extract - 25%
2 Lemongrass oil 3.2%
3 Orange Oil 6%
4 Cypress Oil 3%
5 Ginger Oil 0.2%
6 Eucalyptus Oil 10%
7 Tocopherol 0.20%
8 Stearic acid 16%
9 Emulsifying Wax 7.8%
10 Carnauba Wax 6.8%
11 Jojoba Oil 8%
12 Aloe-vera L/E 5%
13 Ethanol 3%
14 Sorbitol 3%
15 Glycerin 6%
16 Na-Methyl Paraben Quantity S.
17 Phenoxy-ethanol Quantity S.
18 Distilled Water Quantity S.

METHOD OF PREPARATION:
1. In one container, Dissolve Aloe Vera Extract/Gel and Luteolin in same with Ethanol and half quantity of water.
2. In another Container, Dissolve selected Emulsifier with continuous temperature and stir until no lumps observed.
3. Mix Step 1 and Step 2 with addition of stearic acid and Tocopherol (Vitamin-E) until it becomes liquid with smooth texture.
4. Mix all other essential oils, carrier oil and preservative in above step 3.
5. Dissolve and make proper liquid phase in emulsifier tank by mixing water phase and remaining quantity of water.
6. Mix Step 4 in Step 5.
7. Continue to mix until it become proper blend and consistent Cream.
Cream base formulations are easy to spread and remove. Cream base drug delivery system with proper excipients and manufacturing process is elegant drug delivery system. Cream base drug delivery system is pleasing both in appearance and feel post application also. Creams are not greasy and easy to rinse. They are very good for the most of the topical purposes and are considered particularly suited for application on skin portion and inflamed sites also. The present invention is formulated in cream base and hence most preferred drug delivery system for aforesaid herbal composition.
Any methods of delivery and/or administration of present application is considered within the scope of this invention, including for example and not by way of limitation, tablet, capsule, powder, granule, microgranule, pellet, soft gel, controlled release form, liquid, Solution, elixir, syrup, Suspension, emul Sion, magma, gel, cream ointment, lotion, transdermal. Sub lingual, ophthalmic, nasal, otic, aerosol, inhalation, spray, parenteral, Suppository and the like. In Suitable cases, Luteolin/Ashwagandha (Withanolides or Withaferin) may be administered by intravenous or intra-arterial infusion. Compositions of the present invention may also be administered in nutraceutical or functional foods. In addition the effective amount of present application may be combined with amino acids, botanicals, functional foods, herbals, nucleotides, nutraceuticals, pharmaceuticals, proteins, and/or vitamins in an effort to enhance the targeted activity.

STABILITY STUDY OF CREAM FORMULATION:
Storage stability study has been conducted for present cream formulation comprising aforesaid composition. The results are generated for exemplified combinations are provided merely as illustrations of the invention and are not intended to be construed as a limitation thereof.
Table 1: Storage stability Study of the cream formulation:


Table 2:
Formulation code Composition of the active ingredients
C1 Luteolin 2%
C2 Luteolin 15%
C3 Luteolin 25%
C4 Luteolin 30%

EFFICACY STUDY OF CREAM FORMULATION:
EXAMPLE: 4

Experimental Design:
Subject Number: 7
Days of application: 3
Applied Quantity: 0.5 to 1 g of each Cream
Method:
The bio-efficacy study of the present cream formulation is done with comparison to marketed cream formulation; and placebo or control with no active ingredients or none of the formulation applied on subject.
Observation method/Conditions:
The study was done based on applying formulation on biceps, triceps and chest region of 7 different age and weight group of human subjects has been done. The purpose of this study was to assess Vasodilation during a single visit using mercury sphygmomanometer this study examined blood pressure at rest, after Chest-Biceps exercise , after applying Marketed Cream during Exercise and Applying Luteolin based Cream during Exercise with interval of 15mins and 30mins.
Table 3:
Formulation Code Composition
F1 (Cream_Present application) No application of cream or cream with no active ingredients.
F2 (marketed preparation) Various ingredient for the same application in market.
F3 (controlled/ Placebo) 25% of Luteolin (1-98%) + other ingredients of formula disclosed in present patent application.

Measurement of Blood Pressure without Exercise and with no application of either Marketed as well as Luteolin Cream of the present invention.
Table 4:
SUBJECT BLOOD PRESSURE

Average Systolic BP
Average Diastolic BP

Sub#1 125 82
Sub#2 120 80
Sub#3 128 85
Sub#4 118 80
Sub#5 124 84
Sub#6 130 85
Sub#7 135 90
880 586
Average Blood Pressure 125.7142857 83.71428571
Table 5:
DAY#1
Average Blood Pressure of subject #1 to #7
Time
Interval After Exercise (F1) Exercise followed by Application of marketed cream (F2) Exercise followed by Application of present patented cream (F3)
Systolic BP Diastolic BP Systolic BP Diastolic BP Systolic BP Diastolic BP
O min 182 88 165 82 157 74
15 min 145 82 148 82 137 76
30 min 137 82 137 76 130 81

DAY#2
Average Blood Pressure of subject #1 to #7
After Exercise (F1) Exercise followed by Application of marketed cream (F2) Exercise followed by Application of present patented cream (F3)
Time
Interval Systolic BP Diastolic BP Systolic BP Diastolic BP Systolic BP Diastolic BP
O min 180 90 160 89 155 82
15 min 150 86 144 83 135 81
30 min 139 83 138 77 129 80

DAY#3
Average Blood Pressure of subject #1 to #7
After Exercise (F1) Exercise followed by Application of marketed cream (F2) Exercise followed by Application of present patented cream (F3)
Time
Interval Systolic BP Diastolic BP Systolic BP Diastolic BP Systolic BP Diastolic BP
O min 186 88 166 82 153 83
15 min 148 82 146 80 137 79
30 min 138 82 134 80 130 80

Note:
Blood pressure was measured by mercury sphygmomanometer with an appropriately sized cuff by a single trained observer who had no knowledge of the biochemical data at the time of the exercise study.
CONCLUSION:
As on the Theory, Arterial blood flow is directly proportional to the Blood Pressure and Vasodilation is the process which increase the blood flow by relaxing the inner walls of arterial blood vessel. From the Theory, previously we conclude that Luteolin have effect on the Vasodilation through supporting NO production and above result we can now conclude that:
Luteolin based Cream when applied during the Heavy Exercise of Biceps, Triceps, Chest- having support of and reduction in Blood pressure with (+ )or (-) 15 mmhg in systolic and Diastolic blood pressure with compare to Blood Pressure after Exercise and (+) or (-) 10mmhg and (+) or (-) 8mmhg Systolic Pressure and Diastolic Pressure respectively from Marketed Cream.

STUDY 2:
EXAMPLE: 5

Experimental Design:
Subject Number: 7
Days of application: 3
Applied Quantity: 0.5 to 1 g of each Cream
Method:
The purpose of this study was to assess vascular function during a single visit using a variety of stimuli to potentially learn more about overall vascular health. Specifically, this study examined blood flow responses at rest and single bout of exercise. Of particular concern were the stability and reliability of the vascular flow responses as assessed with gauge plethysmography. Whereas this study does not offer any hypotheses in terms of the stability or reproducibility of the responses, it is hypothesized that the flow responses will be greater following exercise when compared to rest and by using the Luteolin based cream we can increase the Vasodilation by increasing the Arterial blood flow.
Experimental conditions:
This visit consisted of taking blood flow measurements of the right leg during varying stimuli using strain-gauge plethysmography, with each participant being asked to refrain from exercise and alcohol for twenty-four hours and from food, medication, and caffeine consumption for ten hours. The participant was also asked to bring a pair of shorts to wear during the testing. Prior to each of the vascular stimuli, resting blood flow measures were obtained. Throughout the vascular assessment, the participant remained in a supine position. Prior to any measure, the participant was allowed to relax for 15 minutes. Throughout the experiment, blood pressure and heart rate were monitored. Dynamic exercise, participants were instructed to perform right plantar flexion against a loaded pedal (60% of their maximal voluntary contraction) for fifteen contractions using a one-to-one second cadence.
Protocol:
1. Dynamic plantar flex the foot for 15 contractions (1/1 second flex/rest cadence)
2. During 13th contraction, inflate ankle cuff to 240mmHg.
3. During 15th contraction, inflate rapid thigh cuff to 7mmHg below diastolic, 1% balance plethysmograph, and measure arterial inflow for 10 seconds.
4. Relax thigh cuff for 10 seconds and repeat arterial inflow measurement for 10 seconds. (Repeat for 3rd measurement).
5. Deflate all cuffs.
Measurement of Arterial Blood Flow without Exercise and with no application of either Marketed as well as Luteolin Cream of the present invention.

Table 6:
SUBJECT Arterial Blood Flow (ml/100ml/min)
Sub#1 2
Sub#2 2.5
Sub#3 3.2
Sub#4 1.8
Sub#5 2.8
Sub#6 2.5
Sub#7 2
Total 16.8
Average Arterial Blood flow 2.4

Table 7:
DAY#1
After Exercise (F1) Exercise followed by Application of marketed cream (F2) Exercise followed by Application of present patented cream (F3)
Arterial Blood Flow (ml/100ml/min) of subject #1 to #7 31.4285 35.428 42.142

DAY#2
After Exercise (F1) Exercise followed by Application of marketed cream (F2) Exercise followed by Application of present patented cream (F3)
Arterial Blood Flow (ml/100ml/min) of subject #1 to #7 32.2867 36.569 43.280

DAY#3
After Exercise (F1) Exercise followed by Application of marketed cream (F2) Exercise followed by Application of present patented cream (F3)
Arterial Blood Flow (ml/100ml/min) of subject #1 to #7 32.3280 37.763 43.960

CONCLUSION:
From the above result we can conclude that Luteolin based cream increase the Arterial blood flow (±)10 ml/100ml/min and (±)6 ml/100ml/min with compare to after Exercise only and After applying the Marketed Cream respectively which promote the Vasodilation of Arterial blood Vessel and decreasing the Blood Pressure.
STUDY 3:
EXAMPLE: 6

Experimental Design:
Subject Number: 7
Days of application: 5
Applied Quantity: 0.5 to 1 g of each Cream
Method:
Peripheral arterial tonometry (PAT) measurements were taken of three volunteers according to the following study. This is a non-invasive technique for examining peripheral microvascular endothelial function. Microvascular endothelial function is assessed by the direct measurement of changes in digital (finger) pulse volume following reactive hyperemia. Here we have going to demonstrate the Reactive hyperemia during Exercise and by application of marketed as well as Luteolin based cream.
Observation and Conditions
This visit consisted of taking blood flow measurements of the right leg during varying stimuli using strain-gauge plethysmography, with each participant being asked to refrain from exercise and alcohol for twenty-four hours and from food, medication, and caffeine consumption for ten hours. Six volunteers were chosen from a group prescreened by blood draw and a health questionnaire. Measurements of height, weight, resting heart rate, and two readings of blood pressure were taken. Volunteers with an average resting blood pressure greater than 160/90 were omitted. Blood samples were analyzed for information regarding complete blood count (CBC), glucose level, general liver function, and lipid pane. On study days, the subjects reported in a fasted state between 7:30 and 9:00 a.m. Body weight, resting heart rate, and blood pressure were all re-measured. The subjects that had an average resting blood pressure of greater than 160/90 were excluded from the study. Subjects then lied down on their backs to acclimatize to ambient conditions. After 30 minutes, a non invasive, sterile, single-use finger probe was fitted to a finger and a baseline PAT reading was taken and recorded for 10 minutes. The subjects were then subjected to a reactive hyperemia procedure ("RH") which consisted of a period of blood flow occlusion done by exercising 30 Kg work load then to the measurement arm using an upper arm blood pressure cuff, pressurized to achieve a supra-systolic level (equivalent to about 60mm mercury above systolic blood pressure) for 5 minutes. Then the cuff was released and the data recorded for another five minutes. Following this first reading, subjects applied marketed cream. Over the next six hours, the PAT response was monitored at two, four and six hours. And same thing reschedule on other day to measure the by application of Luteolin based cream.
Measurement of Reactive Hyperemia without Exercise and with no application of either Marketed as well as Luteolin Cream of the present invention.
Table 8:
SUBJECT Average Reactive Hypermia (RH) of last 5 days without exercise with period of 6 hrs observation. Baseline
Sub#1 1.6 1420
Sub#2 1.4 1324
Sub#3 1.1 1533
Sub#4 1.5 1240
Sub#5 1.7 1520
Sub#6 1.4 1322
Sub#7 1.7 1256
Total 10.4 9615
Average 1.485714286 1373.571

Table 9:

Observations Parameters
RH Baseline
Avg. Reactive Hyperemia of Subjects 1.48 1373
Mean reactive Hyperemia of Subjects after Exercise (F1) 1.54 1385
Mean reactive Hyperemia of Subjects after Exercise and soon applying Marketed Cream. (F2) 2.2 1138
Mean reactive Hyperemia of Subjects after Exercise and soon applying Luteolin Cream. (F3) 2.4 1286

CONCLUSION:
From the above result we can conclude that Luteolin based cream increase Reactive hyperemia by 56% and 10% with compare to after Exercise only and after applying the Marketed Cream respectively which promote the Vasodilation of Arterial blood Vessel and decreasing the Blood Pressure.
While the present invention is described above in connection with preferred or illustrative embodiments are not intended to be exhaustive or limiting of the invention, rather, the invention is intended to cover all alternatives, modifications and equivalents included within its scope, as defined by the appended claim.

,CLAIMS:CLAIMS:
We Claim:

1. A Transdermal cream formulation for vasodilation comprising:
i. Luteolin present in an amount of 1-75%;
ii. One or more anti-inflammatory agent;
iii. One or more vasodilator;
iv. One or more antioxidant;
v. One or more antispasmodic agent;
vi. One or more inactive excipients ;
vii. oil phase present in the range of 5-50%;
viii. Aqueous phase present in the range of 61-77%;
wherein Luteolin is extracted from salvia tomentosa (Balsamic Saga) or Apium graveolens (Celery) or Capsicum annum (green pepper) or Thymus vulgaris (Thyme) or Matricaca Chamomilla, Chrysanthemum morifolium or Loricena japonica or mixture thereof.

2. The transdermal cream formulation for vasodilation as claimed in claim 1, wherein anti-inflammatory agent is selected from cypress oil with concentration of 0.5-15% or ginger oil with concentration of 0.1- 8% or lemongrass oil with concentration of 0.1-20% or eucalyptus oil with concentration of 1-25% or mixture thereof.

3. The transdermal cream formulation for vasodilation as claimed in claim 1, wherein antioxidant agent is selected from an extract from plant source including Aloe Vera with concentration of 2.5-10% or tocopherol with concentration of 0.1-5% or mixture thereof.

4. The transdermal cream formulation for vasodilation as claimed in claim 1, wherein antispasmodic agent is an orange oil with concentration of 3-15%.

5. The transdermal cream formulation for vasodilation as claimed in claim 1, wherein the aqueous (water) phase includes glycerin, sorbitol, demineralized water, perfume.

6. The transdermal cream formulation for vasodilation as claimed in claim 1, wherein the oil phase includes stearic acid and jojoba oil.

7. The transdermal cream formulation for vasodilation as claimed in claim 1, wherein the formulation in addition includes one or more inactive excipients selected from the category of emulsifier, permeation enhancer, preservative, humectant, fragrance.

8. The transdermal cream formulation for vasodilation as claimed in claim 7, wherein Emulsifier is selected from stearic acid, potassium stearate, PEG 400 stearate, Glyceryl monostearate, Arlacel 165, sodium stearate, bees wax, carnauba wax, emulsifying wax or mixture thereof.

9. The transdermal cream formulation for vasodilation as claimed in claim 7, wherein permeation enhancer is selected from Ethanol, ginger extract/oil, dimethyl sulfoxide, dimethyl isosorbide, Isopropyl myristate and propylene glycol, eucalyptus oil, jojoba oil, peppermint oil, fennel oil, almond oil or mixture thereof.

10. The transdermal cream formulation for vasodilation as claimed in claim 7, wherein Humectant is selected from Sorbitol or aloe Vera gel or glycerin or xylitol, maltitol, propylene glycol, butylene glycol or mixture thereof.

11. The transdermal cream formulation for vasodilation as claimed in claim 7, wherein preservative is selected from Methylparben, Propyl paraben, Butyl paraben, ethanol, phenoxy ethanol, Benzoic Acid/Sodium Benzoate, citric acid, Sorbic Acid/Potassium sorbate, Levulinic Acid, Anisic Acid or mixture thereof.

Dated this 21st day of June, 2021.