Pharmaceutical formulation with improved skin permeability

Abstract

The present invention discloses a formulation with improved skin permeability. Particularly, the invention is related to a formulation in the form of nanoemulsion comprising at least one lubricant, saponin, oil component, surfactant and at least one pharmaceutical or nutraceutical active ingredient with improved skin permeability.

Description

PHARMACEUTICAL FORMULATION WITH IMPROVED SKIN PERMEABILITYRELATED APPLICATIONThis application claims the priority of Indian Patent Application no. 202424050650 filed on 2ndJuly, 2024.FIELD OF INVENTION

[0001] The Present invention relates to a formulation with improved skin permeability. Particularly, the invention is related to a formulation in the form of nanoemulsion comprising at least one lubricant, saponin, oil component, surfactant and at least one pharmaceutical or nutraceutical active ingredient.BACKGROUND OF THE INVENTION

[0002] Skin is the largest organ of the body, protects humans from microbes, external pathogens, and other external environmental threats. The other role of skin includes regulation of body temperature, stores water and fat, prevents water loss, involved in sensory and motor responses through sensations of touch, heat and cold, and involved in the production of vitamin D. Human skin is made of 70% water, 27.5% protein. 2% fat, 0.5% mineral salts and trace elements.

[0003] Skin has three layers: a) the epidermis (the outermost layer of skin); b) the dermis (beneath the epidermis); and c) the hypodermis (the deeper subcutaneous dermis). The epidermis is the top layer of skin and the thinnest layer of the epidermis, and it contains three types of cells: squamous cells (stratum comeum). basel cells (second layer of epidermis), and melanocytes (base of epidermis). The dermis is the middle layer of the skin containing blood vessels, lymph vessels, hair, follicles, sweat glands, collagen bundles, fibroblasts, nerves, pain and touch receptors, and sebaceous glands. The dermis provides flexibility and strength to the skin and is held together by a protein called collagen. The hypodermis layer is the deepest layer of skin made of collagen and fat cells and acts as a shock absorber.

[0004] The mean thickness of epidermis was found in the range of 76.9 + 26.2 to 267.4 + 120.6 pm. Dermis is the thickest part of the skin. The mean dermal thickness ranged from 2115 + 946.4 to 5888 + 2422.2 pm.

[0005] One of the important functions of skin is to prevent the water loss and provide the barrier for the transport of the molecules. The body would rapidly dehydrate in the absence of skin. Although most substances cannot penetrate the skin, several strategies have been developed to increase the permeability of small molecules and small peptides. Several formulations and techniques are available to increase the permeability of molecules through the skin.

[0006] US5238933A disclose skin permeation enhancer compositions containing a lower aliphatic ester of a lower aliphatic carboxylic acid such as ethyl acetate and a lower alkanol such as propylene glycol which increase the permeability' of skin to transdermally administered pharmacologically active agents.

[0007] US5716638A disclose a liposomal composition for topical application to the skin, resulting in enhanced transdermal passage through or introduction of an active ingredient into the skin, which composition comprises from 0.5% to 10% phospholipids, from 5% to 35% of a C3 or C4 alcohol, 151 to 30% ethanol, yvherein the combined content of ethanol and C3 or C4 alcohol is at least 20 t. % and not more than 40 wt. %, up to 20 wt. % glycol, at least 20% water and at least one active ingredient.

[0008] EP3880173Aldisclsoes a composition for topical, transdermal, intranasal, mucosal or oral application or administration comprising an oil-in-water nanoemulsion, the nanoemulsion comprising: (a) an aqueous phase; (b) at least one oil; (c) at least one quaternary ammonium compound; and (d) at least one nonionic surfactant.

[0009] The permeability of large molecules such as protein through skin is usually difficult. In view of the same, the present inventors felt that there is a scope in the art to provide a composition that would impart significant skin permeability of one or more active agents including large molecules such as proteins. This remains the objective of the invention.SUMMARY OF THE INVENTION

[0010] In accordance with the above, the present invention provides a novel formulation with improved skin permeability comprising;A. An active Pharmaceutical or nutraceutical agent;B. Composition B; wherein the composition B comprises; i. Lubricant in an amount of 0.5 to 20% by weight based on the total weight of composition B; ii. Saponin or its salt thereof in an amount of 50 to 90% by weight based on the total weight of composition B; iii. Cs - C20 fatty acid or a salt thereof; mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof; squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof in an amount of 9.5 to 40 % by weight based on the total weight of composition B.

[0011] The pharmaceutical agent in the formulation is selected from the group consisting of analgesics, antibiotics, anti-infective, antibodies, antiepileptics, antigens from plants, antirheumatics, benzimidazole derivatives, beta-blocker, cardiovascular drugs, chemotherapeutics, CNS drugs, digitalis glycosides, gastrointestinal drugs, enzymes, hormones, oligonucleotides, peptidhormon proteins, therapeutical bacteria, peptides, proteins, urology drugs, vaccines or mixture thereof in suitable amounts.

[0012] The nutraceutical agent is selected from the group consisting of resveratrol from grape products as an antioxidant, soluble dietary fiber products such as psyllium seed husk for reducing hypercholesterolemia, broccoli (sulphane) as a cancer preservative, and soy or clover (isoflavonoids) to improve arterial health, flavonoids, antioxidants, alpha-linoleic acid from flax seed, beta-carotene from marigold petals or antocyanins from berries and such like in suitable amounts.

[0013] The lubricant in composition B is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, retinoids, alpha hydroxyl acid, beta hydroxy acid, resveratrol, stem cell, EGF (epidermal growth factors), KGF (keratinocyte growth factors). FGF (fibroblastgrowth factors), HGH (human growth hormones), niacinamide, aloe vera, allantoin, or a combination thereof.

[0014] The saponins used in the composition B having emulsifying and surfactant properties comprises of the compounds containing a steroid or triterpenoid aglycone linked to one or more sugar moieties.

[0015] The C5 to C20 fatty acid or a salt thereof present in composition B comprises mono-, di-, tri- glycerides of C5 to C20 fatty7acid or a salt thereof selected from a-linolenic acid, stearidonic acid, eicosapentaenoic acid, cervonic acid, linoleic acid, linolelaidic acid, linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid, mead acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, tricaprylin, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, nonadecylic acid, arachidic acid alone or a combination thereof; tall oil and fractions thereof derived from tallow, fish oil. soybean oil, rapeseed oil, sunflower oil. com oil, safflower oil. palm oil. palm kernel oil, and / or fatty acids derived from other vegetable or animal based triglycerides, and / or fatty acids resulting from hydrolysis of fractions of these blends, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof.

[0016] In another aspect, the formulation of the present invention is a topical or transdermal formulation with improved skin permeability.

[0017] In yet another aspect, the formulation of the present invention is in the form of microemulsion.

[0018] Accordingly, the present invention provides a nanoemulsion formulation with improved skin permeability comprising; i. An aqueous component in an amount of 50% to 97 % by weight, based on total weight of nanoemulsion; ii. A surfactant and / or an emulsifier in an amount of 0.5 % to 15 % by weight, based on total weight of nanoemulsion; iii. An oil component in an amount of 2.5 % to 35 % by weight, based on total weight of nanoemulsion;wherein the aqueous component comprises an active Pharmaceutical or nutraceutical agent, lubricant, saponin or its salt thereof; and wherein the oil component comprises C5 - C20 fatty acid or a salt thereof, mono-, di-, triglycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof.

[0019] In another aspect, the present invention provides a method for preparation of a nanoemulsion comprising; i. Preparing an aqueous solution by mixing lubricant, surfactant and / or emulsifier and saponin in water by continuous stirring; ii. Preparing an oil phase; iii. Mixing the oil phase of step (ii) and aqueous solution of step (i) under continuous stirring to obtain an emulsion; wherein the active pharmaceutical or nutraceutical agent is mixed in aqueous solution or oil phase or mixed to nanoemulsion of step (iii) as a solution in a solvent or buffer to obtain a nanoemulsion comprising active pharmaceutical or nutraceutical agent.DESCRIPTION OF THE FIGURES

[0020] FIG.l : Depict the BoNT gene and its associated protein together forming a polycistronic transcription unit. The Fig represents the locus of BoNT gene, NBP, the hemagglutinin gene and the loci of NAPs, and also the locus of transcription regulator for BoNT gene cluster. There are two types of gene cluster. Type I lacks for Orf- (negative) but contains hemagglutinin (HA+). Type II is complementary form of type I.

[0021] FIG. 2: Depicts DAS score of BoNT / A topical formulations. 10, 25, 50 and 100 units of BoNT / A applied topically to the gastrocnemius muscles of the hind leg of each mice. The control was untreated mice and vehicle control was only formulating agent applied topically to the gastrocnemius muscles of the hind leg of each mice. Standard deviation was taken for 5 mice for each experiment.

[0022] FIG. 3: Depicts Recovery' assay of topical formulations (Toe-spread assay). All the mice were applied topically with different units of BoNT / A toxin (25, 50 and 100 units).Formulations were applied to the gastrocnemius muscles of the hind leg of each mice and score were recorded by three independent observers for 10 days (every 24 hr interval).

[0023] FIG. 4: Depicts Recovery assay of topical formulations (Rotarod assay). All the mice were applied topically with different units of BoNT / A toxin (25, 50 and 100 units). Formulations were applied to the gastrocnemius muscles of the hind leg of each mice and score were recorded by three independent observers for 10 days (every 24 hr interval).

[0024] FIG. 5: Depicts DAS- score of toe-spread assay. Representative figure of each DAS score from the experiment, including control, after 48 hr of the topical formulation application on the gastrocnemius muscles.

[0025] FIG. 6: Depicts transport of IgG through human skin.DETAILED DESCRIPTION OF THE INVENTIONThe invention will now be described in its preferred and optional embodiments so that various aspects of the invention will be fully understood and appreciated w ithout limiting the scope of the invention.The formulation / composition as used interchangeably in the entire specification relates to the formulation / composition with improved skin permeability comprising an active pharmaceutical or nutraceutical agent and the composition B.

[0026] In an embodiment, the present invention relates to a formulation with improved skin permeability comprising;A. An active Pharmaceutical or nutraceutical agent;B. a Composition B; wherein the composition B comprises; i. Lubricant in an amount of 0.5 to 20% by weight based on the total weight of composition B; ii. Saponin or a salt or a derivative thereof in an amount of 50 to 90% by weight based on the total weight of composition B; iii. Cs - C20 fatty acid or a salt thereof; mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof; squalene, hydrogenated polyisobutene, microalgal oil, or amixture thereof in an amount of 9.5 to 40 % by weight based on the total weight of composition B.

[0027] In another embodiment, the present invention discloses a formulation with improved skin permeability comprising;A. at least one active pharmaceutical agent or nutraceutical agent; andB. a composition B; wherein the composition B. comprises i. Lubricant in an amount of 0.5 to 20 % by weight, based on total weight of composition B; ii. Saponin or a salt or a derivative thereof in an amount of 50 to 90% by weight, based on total weight composition B., at least saponin or a salt or a derivative thereof; and iii. Cs to C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof in an amount of 9.5 to 30 % by weight, based on total weight of composition B.

[0028] In yet another embodiment, the present invention discloses a composition with improved skin permeability comprising;A. at least one active pharmaceutical agent or nutraceutical agent; andB. a composition B; wherein the composition B. comprises i. Lubricant in an amount of 0.5 to 10 % by weight, based on total weight of composition B; ii. Saponin or a salt or a derivative thereof in an amount of 50 to 90% by weight, based on total weight composition B., at least saponin or a salt or a derivative thereof; and iii. Cs to C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of Cs to C20 fatty7acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof in an amount of 9.5 to 30 % by weight, based on total weight of composition B.

[0029] In another embodiment, the present invention discloses a composition with improved skin permeability comprising;A. at least one active pharmaceutical agent or nutraceutical agent; andB. a composition B; wherein the composition B. comprises i. Lubricant in an amount of 0.5 to 10 % by weight, based on total weight of composition B; ii. Saponin or a salt or a derivative thereof in an amount of 60 to 90% by weight, based on total weight composition B.. at least saponin or a salt or a derivative thereof; and iii. Cs to C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof in an amount of 10 to 30 % by weight, based on total weight of composition B.

[0030] In yet another embodiment, the present invention discloses a composition with improved skin permeability comprising;A. at least one active pharmaceutical agent or nutraceutical agent; andB. a composition B; wherein the composition B. comprises i. Lubricant in an amount of 0.5 to 8 % by weight, based on total weight of composition B; ii. Saponin or a salt or a derivative thereof in an amount of 65 to 85% by weight, based on total weight composition B., at least saponin or a salt or a derivative thereof; and iii. Cs to C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of Cs to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof in an amount of 10 to 30 % by weight, based on total weight of composition B.

[0031] In another embodiment, the present invention discloses a composition with improved skin permeability- comprising;A. at least one active pharmaceutical agent or nutraceutical agent; andB. a composition B; wherein the composition B. comprises i. Lubricant in an amount of 0.5 to 5 % by weight, based on total weight of composition B; ii. Saponin or a salt or a derivative thereof in an amount of 70 to 85% by weight, based on total weight composition B., at least saponin or a salt or a derivative thereof; and iii. C to C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof in an amount of 10 to 30 % by weight, based on total weight of composition B.

[0032] In an embodiment, the composition as described above is used as a topical formulation with improved skin permeability.

[0033] The pharmaceutical agent in the composition is selected from the group consisting of analgesics, antibiotics, anti -infective, antibodies, antiepileptics, antigens from plants, antirheumatics, benzimidazole derivatives, beta-blocker, cardiovascular drugs, chemotherapeutics, CNS drugs, digitalis glycosides, gastrointestinal drugs, enzy mes, hormones, oligonucleotides, peptidhormon proteins, therapeutical bacteria, peptides, proteins, urology drugs, vaccines or mixture thereof.

[0034] The composition of the present invention comprises 0.0001 to 99 % by weight at least one active pharmaceutical agent or nutraceutical agent, more preferably7the composition comprises 0.01 to 99 % by weight at least one active pharmaceutical agent or nutraceutical agent, even more preferably the composition comprises 0. 1 to 99 % by weight at least one active pharmaceutical agent or nutraceutical agent, most preferably the composition comprises 0.0001 to 60 % by weight at least one active pharmaceutical agent or nutraceutical agent, and in particular preferably the composition comprises 0.0001 to 40 %by weight at least one active pharmaceutical agent or nutraceutical agent, each based on the total weight of the composition.

[0035] In another preferred embodiment, the composition comprises 1 to 99.9999 % by weight of the composition B., more preferably the composition comprises 1 to 99.99 by weight of the composition B., even more preferably the composition comprises 1 to 99.9 % by weight of the composition B.. most preferably the composition comprises 40 to 99.9999 % by weight of the composition B., and in particular preferably of the composition comprises 60 to 99.9999 % by weight of the composition B., each based on the total weight of the composition.

[0036] Nutraceuticals can be defined as extracts of foods claimed to have medical effects on human health. The nutraceutical is usually contained in a medical format such as capsule, tablet or powder in a prescribed dose.

[0037] In another embodiment the nutraceutical agent is selected from resveratrol from grape products as an antioxidant, soluble dietary fiber products, such as psyllium seed husk for reducing hypercholesterolemia, broccoli (sulphane) as a cancer preservative, and soy or clover (isoflavonoids) to improve arterial health, flavonoids, antioxidants, alpha-linoleic acid from flax seed, beta-carotene from marigold petals or antocyanins from berries.

[0038] In another preferred embodiment the active pharmaceutical agent is botulinum toxin (BoNTs), or a fragment of BoNTs thereof. In another preferred embodiment the botulinum toxin or the fragment of BoNTs further comprises of other associated proteins of botulinum toxin complex, such as NBP, HA proteins. P-80, or any combination thereof.

[0039] In another preferred embodiment the active pharmaceutical agent is Immunoglobulin G (IgG).Composition B

[0040] In a preferred embodiment the composition B comprises i. 0.5 to 20 % by weight, based on total weight of composition B., at least one lubricant (sodium hyaluronate); ii. 50 to 90% by weight, based on total weight composition B., at least saponin or asalt or a derivative thereof; and iii. 9.5 to 40 % by weight, based on total weight of composition B., at least one Cs to C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof.

[0041] Component i. (Lubricant)In a preferred embodiment the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, retinoids, alpha hydroxyl acid, beta hydroxy acid, resveratrol, stem cell, EGF (epidermal growth factors), KGF (keratinocyte growth factors), FGF (fibroblast growth factors), HGH (human growth hormones), niacinamide, aloe vera, allantoin, or a combination thereof; more preferably the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, retinoids, alpha hydroxyl acid, beta hydroxy acid, resveratrol, EGF (epidermal growth factors), KGF (keratinocyte growth factors), FGF (fibroblast growth factors), HGH (human growth hormones), niacinamide, aloe vera, allantoin, or a combination thereof; even more preferably the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, retinoids, alpha hydroxyl acid, resveratrol, EGF (epidermal growth factors), KGF (keratinocyte growth factors), FGF (fibroblast growth factors), HGH (human growth hormones), niacinamide, aloe vera, allantoin, or a combination thereof; most preferably the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, EGF (epidermal growth factors), KGF (keratinocyte growth factors), FGF (fibroblast growth factors), HGH (human growth hormones), niacinamide, allantoin, or a combination thereof; particular preferably the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, or a combination thereof; even more particular preferably the lubricant is selected from hyaluronic acid or a salt thereof.

[0042] In another preferred embodiment the salt of hyaluronic acid, or salt of hydrolyzed hyaluronic acid or salt of acetylated hyaluronic acid or salt of hydrolyzed acetylated hyaluronicacid is selected form alkali metal salt or alkaline earth metal salt, more preferably the alkali metal salt, more preferably the salt of sodium and potassium, and most preferably the sodium salt of hyaluronic acid or hyaluronic acid derivative.

[0043] The amount of lubricant is present in the composition B. in the range of 0.5 to 20 % by weight, based on total weight of composition B., preferably, in the range of 0.5 to 15 % by weight, based on total weight of composition B., more preferably in the range of 0.5 to 12 % by weight, based on total weight of composition B., even more preferably in the range of 0.5 to 10 % by weight, based on total weight of composition B., and most preferably in the range of 1 to 5 % by weight, based on total weight of composition B.

[0044] Component ii. (Saponin)Saponins are compounds containing a steroid or triterpenoid aglycone linked to one or more sugar moieties. The saponin having steroid part is acidic nature where saponin with triterpenoid part is neutral in nature. The sugar moieties of the saponins are glucose, arabinose, xylose and glucuronic acid. Saponins can be extracted from plants or marine plants. Saponin has emulsifying and surfactant properties.

[0045] The amount of saponin or a salt or a derivative thereof is present in the composition B. in the range of 50 to 90% by weight, based on total weight composition B., more preferably 60 to 90% by weight, based on total weight composition B., even more preferably 65 to 85 % by weight, based on total weight composition B.. most preferably 70 to 85% by weight, based on total weight composition B., and in particular preferably 70 to 80 % by weight, based on total weight composition B.

[0046] Component iii.In another preferred embodiment the composition B. comprises at least one Cs to C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of Cs to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof, more preferably at least one C5 to C20 fatty acid or a salt thereof, or mono-, di-, tri- glycerides of Cs to C20 fatty acid or a salt thereof, or a mixture thereof.

[0047] In another preferred embodiment the Cs to C20 Patty acid or a salt thereof, mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof is selected from a-linolenic acid, stearidonic acid, eicosapentaenoic acid, cervonic acid, linoleic acid, linolelaidic acid, linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid, mead acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, tricaprylin, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, nonadecylic acid, arachidic acid, or salts thereof, mono-, di-, tri- glycerides of Cs to C20 fatty acid or a salt thereof, or a combination thereof, tall oil and fractions thereof, derived from tallow, fish oil, soybean oil, rapeseed oil, sunflower oil , com oil, safflower oil, palm oil, palm kernel oil, and / or fatty acids derived from other vegetable or animal based triglycerides, and / or fatty acids resulting from hydrolysis of fractions of these blends.

[0048] In another preferred embodiment the mono-, di-, tri- glycerides of C5 to C20 fatty’ acid are glycerol esters of at least one fatty’ acid selected from linolenic acid, stearidonic acid, eicosapentaenoic acid, cervonic acid, linoleic acid, linolelaidic acid, linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid, mead acid, valeric acid, caproic acid, enanthic acid, capry lic acid, pelargonic acid, capric acid, tricaprylin, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, nonadecylic acid, arachidic acid, or a combination thereof. Preferably the mono-, di-, tri- glycerides of C5 to C20 fatty' acid are glycerol esters of at least one fatty acid selected from linolenic acid, linoleic acid, linolelaidic acid, linolenic acid, palmitoleic acid, oleic acid, elaidic acid, valeric acid, caproic acid, caprylic acid, capnc acid, tri caprylin. lauric acid, myristic acid, palmitic acid, stearic acid, or a combination thereof, more preferably the mono-, di-, tri- glycerides of C5 to C20 fatty' acid are glycerol esters of at least one fatty’ acid selected from linolenic acid, linoleic acid, linolelaidic acid, linolenic acid, palmitoleic acid, oleic acid, caproic acid, capry lic acid, capric acid, tricaprylin, lauric acid, myristic acid, palmitic acid, stearic acid, or a combination thereof, even more preferably the mono-, di-, tri- glycerides of C5 to C20 fatty acid are glycerol esters of at least one fatty7acid selected from palmitoleic acid, oleic acid, caproic acid, caprylic acid, capric acid, tricaprylin, lauric acid, myristic acid, palmitic acid, stearic acid, or a combination thereof, and most preferably the mono-, di-, tri- glycerides of Cs to C20 fatty acid are glycerolesters of at least one fatty acid selected from caproic acid, caprylic acid, capric acid, tri caprylin, or a combination thereof

[0049] In another preferred embodiment, the fatty acids are obtained from vegetable or animal fats and oils.

[0050] The amount of component (iii) is present in the composition B. in the range of 9.5 to 40% by weight, based on total weight composition B., more preferably 10 to 35% by weight, based on total weight composition B, even more preferably 10 to 30 % by weight, based on total weight composition B, most preferably 10 to 55% by weight, based on total weight composition B, and in particular preferably 15 to 25 % by weight, based on total weight composition B.

[0051] In another preferred embodiment the ratio of the component (i) to the component (ii) in the composition B is in the range of 1 :20 to 1 :50, more preferably the ratio of the component(i) to the component (ii) in the composition B is in the range of 1:25 to 1 :50, even more preferably the ratio of the component (i) to the component (ii) in the composition B is in the range of 1 :25 to 1 :40, most preferably the ratio of the component (ii) to the component (i) in the composition B is in the range of 1 :30 to 1 :40, and in particular the ratio of the component(ii) to the component (i) in the composition B is in the range of 1 :32 to 1 :40. In an embodiment for 1 part of component (i) preferably 20 to 50 parts of component (ii), more preferably for 1 part of component (i) 20 to 45 part of component (ii), even more preferably for 1 part of component (i) 25 to 45 part of component (ii), most preferably for 1 part of component (i) 30 to 45 part of component (ii), and particular preferably for 1 part of component (i) 30 to 40 part of component (ii) each in composition B, or the pharmaceutical composition, or even in the nanoemulsion as described in the present invention.

[0052] It has been identified that the above ratio is particularly beneficial for improved skin permeability of the active pharmaceutical and / or nutraceutical agent.

[0053] Further components:In another preferred embodiment the composition further comprises at least one surfactant,emulsifier, preservative, humectants, emollients, stabilizer, pH modifier or a combination thereof.

[0054] Surfactant / emulsifierSurfactants are amphiphilic, meaning that they contain hydrophilic (water-loving) head groups and hydrophobic (water-hating, or oil-loving) tails. An emulsifier is a surfactant that stabilizes emulsions. Thus, in another preferred embodiment the surfactant and emulsifier may be the same. In another preferred embodiment the surfactant and / or the emulsifier in the composition B is different from component a. and component b.

[0055] The at least one emulsifier is preferably selected from polyglycosides, alcohols, sugar and sugar derivatives, polyethers, amines, polyethylene derivatives, alkyl sulfates (e.g. sodium dodecyl sulfate), alky l ether sulfates, dioctyl sodium sulfosuccinate, polysorbates (e.g. polyoxyethylene (20) sorbitan monooleate), nonylphenol ethoxylates (nonoxynol-9) and mixtures thereof. The at least one emulsifier is preferably selected from alkyl polyglycosides, decyl glucoside, decyl polyglucose, lauryl glucoside, octyl glucoside, N-octyl beta-D- thioglucopyranoside, cetostearyl alcohol, cetyl alcohol, stearyl alcohol, polyoxyethylene cetostearyl alcohol, cetylstearyl alcohol, oleyl alcohol, polyglyceryl-6-dioleate, glyceryl stearate citrate, polyglyceryl-3 caprate, polyglyceryl-3 diisostearate, glyceryl isostearate, polyglyceryl-4-isostearate, glyceryl monolinoleate, dicaprylyl carbonate, alcohol polyglycol ether, polyethylenglycolether of cetearylalcohols (n=20), polyethylene glycol-6 stearate, glycol stearate, polyethylene glycol-32 stearate, polyethylene glycol-20 stearate, fatty alcohol polyglycol ether, polyethylene glycol-4 laurate, polyethylene glycol isocetyl ether (n=20), poly ethylenegly col-32 (Mw 1500 g / mol) mono- and diesters of lauric acid (C12), nonaethylene glycol, polyethylene glycol nonylphenyl ether, octaethylene glycol monododecyl ether, pentaethylene glycol monododecyl ether, polyethylene glycol macrocetyl ether, polyethylene glycol esters of palmitic (C16) or stearic (C18) or caprylic acids, polyoxyethylene fatty ether derived from stearyl alcohols like BRU S2, polyoxyethylene oxypropylene stearate, macrogol stearyl ether (20), diethylaminoethyl stearate, polyethylene glycol stearate, sucrose distearate, sucrose tristearate, sorbitan monostearate, sorbitan tristearate, mannide monooleate, octaglycerol monooleate, sorbitan dioleate, polyricinoleate, polysorbate like polysorbate 20 and Polyoxyethylene (20) sorbitan monooleate (polysorbate 80). sorbitan, sorbitanmonolaurate, sucrose cocoate, glycereth-2 cocoate, ethylhexyl cocoate, polypropylene glycol - 3 benzyl ether myristate, sodium myristate, gold sodium thiomalate, polyethylene glycol 8 laurate, polyethylene-4 dilaurate, from a- Hexadecyl-cj-hydroxypoly(oxyethylene), cocamide diethanolamine, N-(2- hydroxyethyl)dodecanamide, octylphenoxypolyethoxyethanol. maltoside, 2,3-Dihydroxypropyl dodecanoate, 3-[(3R,6R,9R,12R,15S,22S,25S,30aS)- 6,9,15,22-Tetrakis(2-anhno-2-oxoethyl)-3-(4-hydroxybenzyl)-I2-(hydroxymethyl)-18-(l l- methyltridecyl)-! 4, 7. 10. 13. 16, 20,23,26- nonaoxotriacontahydropyrrolofl .2-g] [ 1 4, 7. 10. 13, 16, 19, 22,25] nonaazacyclooctacosin-25-yljpropenamide, 2-{2-[2-(2-{2-[2-(2-{2-[2-(4- nony 1 phenoxy )ethoxy] ethoxy } ethoxy )ethoxy] ethoxy } ethoxy)ethoxy ] ethoxy } ethanol, oxypolyethoxy dodecane, poloxamers like pol oxamer 188 (Pluronic F-68) and poloxamer 407, propylene glycol monocaprylate, type I (Capryol PGMC). polyethoxylated tallow amine, polyglycerol, polyoxyl 40 hydrogenated castor oil, surfactin, 2-[4-(2,4,4-trimethylpentan-2- yl)phenoxy] ethanol, carbomer, sodium carbomer, carboxymethylcellulose calcium, carrageenan, cholesterol, deoxycholic acid, phospholipids like egg phospholipids, gellan gum, lanolin, capric acid, waxes like Polawax NF, Polawax A31 or Ceral PW. ester gum, dea-cetyl phosphate, soya lecithin, sphingomyelins, sodium phosphate, sodium lauroyl lactylate, lanolin. Oxirane methyl- polymer with oxirane monobutyl ether, 1 ,2-dierucoylphosphatidylcholine, dimethicone end-blocked with an average of 14 moles of propylene oxide, laurylmethicone copolyol, lauroglycol 90, white mineral oil like Amphocerine KS, dispersion of acrylamide / sodium acryloyldimethyl taurate copolymer in isohexadecane, and sodium polyacrylate or mixtures thereof.

[0056] In another preferred embodiment the amount of surfactant and / or emulsifier in composition B. is present in an amount in the range of 0.5 to 50% by weight, based on total weight of thee composition B, more preferably in an amount in the range of 1 to 50% by weight, based on total weight of thee composition B, even more preferably in an amount in the range of 1 to 40% by weight, based on total weight of thee composition B. most preferably in an amount in the range of 1 to 30% by weight, based on total weight of thee composition B, and in particular in an amount in the range of 5 to 20% by weight, based on total weight of thee composition B.

[0057] PreservativeIn another preferred embodiment the preservative is selected from the group consisting ofcetylpyridinium chloride, benzalkonium chloride, benzyl alcohol, chlorhexidine, imidazolidinyl urea, phenol, potassium sorbate, benzoic acid, bronopol, chlorocresol, paraben esters, phenoxyethanol, sorbic acid, alpha-tocophemol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, sodium ascorbate, sodium metabisulphite, citric acid, edetic acid, semi-synthetic derivatives thereof, Other suitable preservatives include, but are not limited to, benzyd alcohol, chlorhexidine (bis (p- chlorophenyldiguanido) hexane), chlorphenesin (3-(-4-chloropheoxy)-propane-1.2-diol). Kathon CG (methyl and methylchloroisothiazolinone), parabens (methyl, ethyl, propyl, butyl hydrobenzoates), phenoxyethanol (2- phenoxyethanol), sorbic acid (potassium sorbate, sorbic acid), Phenonip (phenoxyethanol, methyl, ethyl, buty l, propyl parabens), Phenoroc (phenoxyethanol 0.73%, methyl paraben 0.2%, propyl paraben 0.07%). Liquipar Oil (isopropyl, isobuty l, butylparabens), Liquipar PE (70% phenoxyethanol, 30% liquipar oil), Nipaguard MPA (benzyl alcohol (70%), methyl & propyl parabens), Nipaguard MPS (propylene glycol, methyl & propyl parabens), Nipasept (methyl, ethyl and propyl parabens), Nipastat (methyl, butyl, ethyl and propyel parabens), Elestab 388 (phenoxyethanol in propylene glycol plus chlorphenesin and methylparaben), and Killitol (7.5% chlorphenesin and 7.5% methyl parabens), and combinations thereof;

[0058] In another preferred embodiment the amount of preservative in composition B. is present in an amount in the range of 0.5 to 30% by weight, based on total weight of thee composition B, more preferably in an amount in the range of 1 to 20 % by weight, based on total weight of thee composition B, even more preferably in an amount in the range of 1 to 15 % by weight, based on total weight of thee composition B, most preferably in an amount in the range of 1 to 10 % by weight, based on total weight of thee composition B, and in particular in an amount in the range of 2 to 8 % by weight, based on total weight of thee composition B.

[0059] In another aspect, the formulation of the present invention is a topical or transdermal formulation with improved skin permeability.

[0060] In yet another aspect, the formulation of the present invention is in the form of nanoemulsion.

[0061] Accordingly, the present invention discloses a nanoemulsion formulation with improved skin permeability comprising;i. An aqueous component in an amount of 50% to 97 % by weight, based on total weight of nanoemulsion; ii. A surfactant and / or an emulsifier in an amount of 0.5 % to 15 % by weight, based on total weight of nanoemulsion; iii. An oil component in an amount of 2.5 % to 35 % by weight, based on total weight of nanoemulsion; wherein the aqueous component comprises an active Pharmaceutical or nutraceutical agent, lubricant, saponin or its salt or derivative thereof; and wherein the oil component comprises C5 - C20 fatty acid or a salt thereof, mono-, di-, triglycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof.

[0062] In another preferred embodiment the aqueous component comprises at least one lubricant, at least saponin or a salt or a derivative thereof. In another preferred embodiment the at least one active pharmaceutical agent or nutraceutical agent, surfactant and / or emulsifier may be mixed with the aqueous component.

[0063] The aqueous component may further comprise a cosolvent, preferably an organic solvent, more preferably water miscible organic solvent.

[0064] In another preferred embodiment the nanoemulsion comprises 65 to 95 % by weight of solvent based on total weight of the nano emulsion, preferably 70 to 95 % by weight of solvent based on total weight of the nano emulsion, even more preferably 75 to 95 % by weight of solvent based on total weight of the nano emulsion, most preferably 80 to 95 % by weight of solvent based on total weight of the nano emulsion, and in particular 85 to 95 % by weight of solvent based on total weight of the nano emulsion. In each case the preferred solvent is water.

[0065] In another preferred embodiment the oil component comprises at least one C; to C20 fatty acid or a salt thereof, or mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof.

[0066] In another preferred embodiment, the at least one active pharmaceutical agent or nutraceutical agent, may be mixed with the oil component.

[0067] In another preferred embodiment the nanoparticles comprises in the nanoemulsion have an average particle size distribution of 0.5 to 500 nm, more preferably the nanoparticles comprises in the nanoemulsion have an average particle size distribution of 0.5 to 100 nm, even more preferably the nanoparticles comprises in the nanoemulsion have an average particle size distribution of 0.5 to 50 nm, most preferably the nanoparticles comprises in the nanoemulsion have an average particle size distribution of 1 to 25 nm. and in particular preferably the nanoparticles comprises in the nanoemulsion have an average particle size distribution of 1 to 50 nm.

[0068] In another embodiment, the nanoparticles in the nanoemulsion have a zeta potential of 20 to 38.6 mV.

[0069] In another embodiment, the hydrophilic-lipophilic ratio in the nanoemulsion is in the range of 10:0.5 to 40: 1.

[0070] In another embodiment, the nanoemulsion is an oil-in-water-type emulsion.

[0071] In another embodiment, the pharmaceutical agent in the aqueous component of said nanoemulsion is selected from the group consisting of analgesics, antibiotics, anti-infective, antibodies, antiepileptics, antigens from plants, antirheumatics, benzimidazole derivatives, beta-blocker, cardiovascular drugs, chemotherapeutics, CNS drugs, digitalis glycosides, gastrointestinal drugs, enzymes, hormones, oligonucleotides, peptidhormon proteins, therapeutical bacteria, peptides, proteins, urology drugs, vaccines or mixture thereof. The amount of the pharmaceutical agent will vary depending on the active, its target, its site of action and its solubility and bioavailability.

[0072] The nutraceutical agent in the aqueous component of said nanoemulsion is selected from the group consisting of resveratrol from grape products as an antioxidant, soluble dietary fiber products such as psyllium seed husk for reducing hypercholesterolemia, broccoli (sulphane) as a cancer preservative, and soy or clover (isoflavonoids) to improve arterial health, flavonoids, antioxidants, alpha-linoleic acid from flax seed, beta-carotene from marigold petals or antocyanins from berries and such like in suitable amount.

[0073] In an embodiment, the pharmaceutical agent in the nanoemulsion composition contains pure botulinum toxin A (150 kDa; BoNT / A).

[0074] Botulinum neurotoxin is a large protein toxin (approximately 150 kDa) that is able to bind and internalize to motor neurons very specifically. BoNTs are produced by Clostridium botulinum along with several neurotoxins associated with polypeptides (proteins) (NAPs). The toxin with NAPs is referred herein as a “complex toxin” as disclosed in Fig 1. The BoNT gene and its associated protein together form a polycistronic transcription unit. The Fig 1 represents the locus of BoNT gene, NBP, the hemagglutinin gene and the loci of NAPs, and also the locus of transcription regulator for BoNT gene cluster. There are two types of gene cluster. Type I lacks for Orf- (negative) but contains hemagglutinin (HA+). Type II is a complementary form of type I.

[0075] In an embodiment, the Botulinum neurotoxin may be used alone or in combination with another pharmaceutical agent or nutraceutical agent as described above.

[0076] In another embodiment, the pharmaceutical agent in the nanoemulsion composition contains antibody selected from Immunoglobulin G (IgG).

[0077] IgG antibodies are large globular proteins made of four peptide chains, two identical y (gamma) heavy chains of about 50 kDa and two identical light chains of about 25 kDa. Thus, the total molecular weight of the resulting tetrameric quaternary structure is about 150 kDa.

[0078] The lubricant in the aqueous component of said nanoemulsion is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, retinoids, alpha hydroxyl acid, beta hydroxy acid, resveratrol, stem cell. EGF (epidermal growth factors), KGF (keratinocyte growth factors), FGF (fibroblast growth factors), HGH (human growth hormones), niacinamide, aloe vera, allantoin, or a combination thereof.

[0079] In an embodiment, the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, or a mixture thereof. Preferably the lubricant is sodium hyaluronate in an amount of 0.5 to 10% by weight.

[0080] The saponins in the aqueous component of said nanoemulsion having emulsifying and surfactant properties comprises of the compounds containing a steroid or triterpenoid aglycone linked to one or more sugar moieties.

[0081] In an embodiment, the pharmaceutical agents, nutraceutical agents, lubricants, saponins in the aqueous component of the nanoemulsion together are present in an amount of 50% to 97% by weight, based on total weight of nanoemulsion.

[0082] The C5 to C20 fatty7acid or a salt thereof in the aqueous component of said nanoemulsion comprises mono-, di-, tri- glycerides of C5 to C20 fatty' acid or a salt thereof selected from a-linolenic acid, stearidonic acid, eicosapentaenoic acid, cervonic acid, linoleic acid, linolelaidic acid, linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid, mead acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, tricaprylin, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, nonadecylic acid, arachidic acid alone or a combination thereof; tall oil and fractions thereof derived from tallow, fish oil, soybean oil, rapeseed oil, sunflower oil, com oil, safflower oil, palm oil, palm kernel oil, and / or fatty' acids derived from other vegetable or animal based triglycerides, and / or fatty acids resulting from hydrolysis of fractions of these blends, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof.

[0083] In an embodiment, the nanoemulsion may further contain surfactant, emulsifier, preservative, humectants, emollients, stabilizer and such like alone or combination thereof in suitable amounts.

[0084] The surfactant is selected from sorbitan fatty acid esters such as polyoxyethylene sorbitan and its monolaurate and monooleates (e.g.. Tween 20, Tween 60, or Tween 80). sorbitan palmitate, oleate, and stearates (e g., Span 40, Span 60, Span 65, or Span 80); polyoxyethylene esters; polyethylene glycol fatty7acid esters such as Cremophor™; diethylene glycol monolaurate, triethanolamine oleate, ethyl laurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 188 alone or combination thereof.

[0085] The emulsifier is selected from polyglycosides, alcohols, sugar and sugar derivatives, polyethers, amines, polyethylene derivatives, alkyl sulfates (e.g. sodium dodecyl sulfate), alkyl ether sulfates, dioctyl sodium sulfosuccinate, polysorbates (e.g. polyoxyethylene (20) sorbitanmonooleate), nonylphenol ethoxylates (nonoxynol-9), alkyl polyglycosides, decyl glucoside, decyl polyglucose, lauryl glucoside, octyl glucoside, N-octyl beta-D-thioglucopyranoside, cetostearyl alcohol, cetyl alcohol, stearyl alcohol, polyoxyethylene cetostearyl alcohol, cetylstearyl alcohol, oleyl alcohol, poly gly ceryl -6-dioleate, glyceryl stearate citrate, polyglyceryl-3 caprate, polyglyceryl-3 diisostearate, glyceryl isostearate, polyglyceryl-4- isostearate. glyceryl monolinoleate, dicaprylyl carbonate, alcohol polyglycol ether, polyethylenglycolether of cetearylalcohols (n=20), polyethylene glycol-6 stearate, glycol stearate, polyethylene glycol-32 stearate, polyethylene glycol-20 stearate, fatty alcohol polyglycol ether, polyethylene glycol-4 laurate, polyethylene glycol isocetyl ether (n=20), polyethyleneglycol-32 (Mw 1500 g / mol) mono- and diesters of lauric acid (C12), nonaethylene glycol, polyethylene glycol nonylphenyl ether, octaethylene glycol monododecyl ether, pentaethylene glycol monododecyl ether, polyethylene glycol macrocetyl ether, polyethylene glycol esters of palmitic (Cl 6) or stearic (Cl 8) or caprylic acids, polyoxyethylene fatty ether derived from steary l alcohols like BRIJ S2, polyoxyethylene oxypropylene stearate, macrogol stearyl ether (20), diethylaminoethyl stearate, polyethylene glycol stearate, sucrose distearate, sucrose tristearate, sorbitan monostearate, sorbitan tristearate, mannide monooleate, octaglycerol monooleate, sorbitan dioleate, polyricinoleate, polysorbate like polysorbate 20 and Polyoxyethylene (20) sorbitan monooleate (polysorbate 80), sorbitan, sorbitan monolaurate, sucrose cocoate, glycereth-2 cocoate, ethylhexyl cocoate, polypropylene glycol - 3 benzyl ether myristate, sodium myristate, gold sodium thiomalate, polyethylene glycol 8 laurate, polyethylene-4 dilaurate, from a- Hexadecyl-cj-hydroxypoly(oxyethylene), cocamide diethanolamine, N-(2- hydroxyethyl)dodecanamide, octylphenoxy polyethoxyethanol, maltoside, 2,3-Dihydroxypropyl dodecanoate, 3-[(3R,6R,9R,12R,15S,22S,25S,30aS)- 6,9,15,22-Tetrakis(2-amino-2-oxoethyl)-3-(4-hydroxybenzyl)-12-(hydroxyl methyl)- 18-(l 1- methyltridecyl)-! ,4,7,10,13, 16,20,23,26- nona oxo triaconta hydropyrrolo[l,2-g][l ,4,7,10,13,16,19,22,25]nonaazacyclooctacosin-25- yl jpropenamide, 2-{2-[2-(2-{2-[2-(2-{2- [2-(4-nonylphenoxy)ethoxy] ethoxy} ethoxy) ethoxy] ethoxy} ethoxy )ethoxy] ethoxy} ethanol, oxy poly ethoxy dodecane, poloxamers like poloxamer 188 (Pluronic F-68) and poloxamer407, propylene glycol monocaprylate type I (Capryol PGMC). polyethoxylated tallow amine, polyglycerol, polyoxyl 40 hydrogenated castor oil, surfactin, 2-[4-(2,4,4-trimethylpentan-2- yl)phenoxy] ethanol, carbomer, sodium carbomer, carboxymethylcellulose calcium, carrageenan, cholesterol, deoxycholic acid, phospholipids like egg phospholipids, gellan gum, lanolin, capric acid, waxes like Polawax NF, Polawax A31 or Ceral PW. ester gum, dea-cetyl phosphate, soya lecithin, sphingomyelins, sodium phosphate, sodium lauroyl lactylate, lanolin,Oxirane methyl- polymer with oxirane monobutyl ether, 1,2-dierucoylphosphatidylcholine, dimethicone end-blocked with an average of 14 moles of propylene oxide, laurylmethicone copolyol, lauroglycol 90, white mineral oil like Amphocerine KS, dispersion of acrylamide / sodium acryloyldimelhyl taurate copolymer in isohexadecane, and sodium polyacrylate or mixtures thereof. Preferred are macrogol stcaryl ether (20) and polysorbate 80, or a mixture thereof.

[0086] The preservatives are selected from the group consisting of cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, phenoxyethanol, ethylhexylglycerol, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate calcium propionate alone or mixtures thereof.

[0087] In an embodiment, the amount of fatty acids present in the oil component is in the range of 9.5 to 40% by weight, based on total weight of the nanoemulsion, more preferably 10 to 35% by weight, even more preferably 10 to 30 % by weight, most preferably 10 to 55% by weight, and in particular preferably 15 to 25 % by weight.

[0088] In another embodiment, the ratio of lubricants to saponins in the aqueous phase is in the range of 1:20 to 1:50, more preferably in the ratio of 1 :25 to 1:50, even more preferably in the ratio of 1 :25 to 1 :40, most preferably in the ratio of 1 :30 to 1:40, and in particular in the ratio of 1:32 to 1:40.

[0089] In another embodiment for 1 part of lubricant preferably 20 to 50 parts of saponin, more preferably for 1 part of lubricant 20 to 45 part of saponin, even more preferably for 1 part of lubricant 25 to 45 part of saponin; most preferably for 1 part of lubricant 30 to 45 part of saponin and particular preferably for 1 part of lubricant 30 to 40 part of saponin are present each in the aqueous phase of the nanoemulsion composition of the present invention.

[0090] In an embodiment, the said ratio of lubricants to saponins in the pharmaceutical composition and / or in nanoemulsion provides improved skin permeability of the active pharmaceutical and / or nutraceutical agent.

[0091] In yet another embodiment, the surfactant and / or emulsifier in the pharmaceutical composition and / or nanoemulsion is present in an amount in the range of 0.5 to 50% by weight,based on total weight of the composition B or the nanoemulsion, more preferably in an amount in the range of 1 to 50% by weight, even more preferably in an amount in the range of 1 to 40% by weight, most preferably in an amount in the range of 1 to 30% by weight, and in particular in an amount in the range of 5 to 20% by weight.

[0092] In another embodiment, the amount of preservative in the pharmaceutical composition and / or nanoemulsion is in an amount in the range of 0.5 to 30% by weight, more preferably in an amount in the range of 1 to 20 % by weight, even more preferably in an amount in the range of 1 to 15 % by weight, most preferably in an amount in the range of 1 to 10 % by weight, and in particular in an amount in the range of 2 to 8 % by weight, based on total weight of thee composition B in the pharmaceutical composition and / or the total weight of the nanoemulsion.

[0093] In another embodiment, the aqueous component in the nano emulsion may further comprise a co-solvent, preferably an organic solvent, more preferably water miscible organic solvent.

[0094] The nanoemulsion comprises 65 to 95 % by weight of solvent based on total weight of the nanoemulsion, preferably 70 to 95 % by weight; more preferably 75 to 95 % by weight of solvent, most preferably 80 to 95 % by weight of solvent, and in particular 85 to 95 % by weight of solvent based on total weight of the nanoemulsion. In each case the preferred solvent is water.

[0095] In an embodiment, the present invention relates to a process for preparation of the formulation / nanoemulsion, comprising the steps: i. Preparing an aqueous solution by mixing lubricant, surfactant and / or emulsifier and saponin in water by continuous stirring; ii. Preparing an oil phase; and iii. Mixing oil phase into aqueous solution under continuous stirring to obtain an emulsion; wherein the active pharmaceutical or nutraceutical agent dissolved in a solvent or buffer is mixed in aqueous solution or oil phase or mixed to nanoemulsion of step (iii) obtain the desired product.

[0096] In an alternate embodiment of the present process, the aqueous phase comprising lubricant, surfactant and / or emulsifier and saponin may be lyophilized to obtain a powderwhich was then mixed in the oil phase and the active ingredients to obtain the product in gel form.

[0097] In yet another embodiment, the nanoemulsion formulation of the present invention is stable at a pH in the range of 5.5 to 8.0.

[0098] In an embodiment, the present invention relates to a pharmaceutical formulation / nanoemulsion comprising BoNT / A as active ingredient as disclosed in table 1 below.

[0099] The BoNT / A pure toxin was dissolved into the formulation according to present invention and was applied on gastrocnemius muscle of mice. Three concentrations were used; 10, 25, 50 and 100 units (1 unit = 1 LD50 = 25 pg for Balb c mice) of pure BoNT / A. The said formulation was capable of delivering 150 kDa molecules through the skin without any aid of chimerical, mechanical or electrical enhancement technique as shown in the figs 2 to 5.

[0100] The present formulation / nanoemulsion is the first of its kind which allows permeation of 150 kDa protein BoNT through the skin without any stimulation (chemical, mechanical or electrical or using ultrasound). Skin is not permeable to large molecules even small molecules are difficult to penetrate. The proposed formulation provides the skin permeation of such large protein molecule without any stimulation.

[0101] In an embodiment, the formulation / nanoemulsion with improved skin permeability comprises BoNT / A pure toxin as pharmaceutical agent in the concentration range of 25- 100LJnits / 20mcl with no toxicity and with the DAS score of 0-3. [DAS score is based on Toespread assay. It is not about lethality assay. It is about active toxin going to the nerve of toes to paralyze when administered topically],

[0102] In an embodiment, the present invention relates to a pharmaceutical formulation / nanoemulsion comprising Immunoglobulin G (IgG) as active ingredient as disclosed in table 2 below.

[0103] In another embodiment, the present invention relates to the method of treating, preventing, inhibiting, delaying onset of, or causing regression of one or more neuro-muscular diseases and conditions or treatment of wrinkle or used in cosmetic treatment, diseases orconditions which include neuronal regeneration / sprouting, disease involving muscle movement, various wounds, scars and gastrointestinal symptoms comprising applying said pharmaceutical composition / nanoemulsion of the present invention.

[0104] In another embodiment, the present invention relates to the formulation / nanoemulsions with improved skin permeability for the treatment, prevention, inhibition, delaying onset of, or causing regression of one or more neuro-muscular diseases and conditions or treatment of wrinkle or used in cosmetic treatments. In some variations the diseases or conditions include neuronal regeneration / sprouting, disease involving muscle movement, various wounds, scars and gastrointestinal symptoms.

[0105] Experimental

[0106] Example 1: Table 1: Nano emulsion Composition*protein used is botulinum toxinTable 2: Nano emulsion Composition*antibody used is IgG

[0107] Example 2: Preparation of NanoemulsionThe nano emulsion was prepared by the general steps as follows: a) Mixing sodium hyaluronate, propylene glycol, Tween-80 and saponin in water by continuous stirring to obtain the aqueous phase containing surfactant; b) Mixing phenoxy ethanol, and caprylic acid / capric triglyceride optionally with hydrogenated castor oil to obtain an oil phase; andc) Adding oil phase into aqueous phase under continuous stirring for 15 min to obtain the nano emulsion without active ingredient; and d) Charging Ing to 10 ng BoNT / A in 10 mM Sodium Phosphate buffer at a pH 7.1 to step c) nanoemulsion at room temperature under continuous stirring to obtain the formulation with active ingredient.Alternately the aqueous phase comprising surfactant may be lyophilized to obtain a powder composition which is then mixed in the oil phase with the active ingredient to obtain the product in gel formulation.

[0108] Example 3: Study

[0109] Example 3A: Preparation of nano-emulsion formulation of pure botulinum toxinA (150 kDa; BoNT / A)

[0110] Nano emulsion was prepared by: a) mixing sodium hyaluronate, propylene glycol, Tween-80 and saponin in water by continuous stirring (solution A); b) either mixed phenoxy ethanol, and carylic acid / capric triglyceride together (without hydrogenated castor oil) or in hydrogenated castor oil (solution B); and c) mixed solution B into solution A by continuous stirring for 15 min (solution C). Prepared the desired concentration of pure BoNT / A in 10 mM Sodium Phosphate buffer, pH = 7.1. Added BoNT / A solution in solution C and stirred the solution at room temperature for 15-20 min (solution D).

[0111] Example 3B: Application of the Formulation / CompositionThe BoNT / A in phosphate buffer formulation was applied topically on gastrocnemius muscle of mice. Three concentrations were used: 25, 50 and 100 units (1 unit = 1 LD50 = 25 pg of Balb C mice) of pure BoNT / A in the above formulation (solution D). For this, Balb C mice were used and housed 5 mice per cage, under 12: 12-h light: dark cycle in paper chip bedding. Water and rodent chow were used for food supplement. Room temperature and humidity w ere maintained between 22 - 26 °C and 50 - 70%, respectively. The toe-spread reflex was observed by lifting the mouse by the base of the tail, with free hind limb, and allowing the forelimbs to rest on the w orkbench. Toe spread was observed for 24 hr to 48 hr. Toe-spread data were analyzed using DAS (digital abduction score) score. Zero DAS score indicates complete spread of toe and DAS score four indicated complete paralysis. Formulations were applied on lefthind limb of the mice and vehicle control (solution C as a control) was applied to the right hind limb of the mice.

[0112] The BoNT / A pure toxin was dissolved into the formulation obtained above according to present invention is applied on gastrocnemius muscle of mice.

[0113] Three concentrations were used; 10. 25. 50 and 100 units (1 unit = 1 LD50 = 25 pg of Balb c mice) of pure BoNT / A.

[0114] For this Balb C mice were used and housed 5 mice per cage, under 12: 12-h light dark cycle in paper chip bedding. Water and rodent chow were used for food supplement. Room temperature and humidity were maintained between 22 - 26 °C and 50 - 70%, respectively. The toe-spread reflex was observed by lifting the mouse by the base of the tail, with free hind limb, and allowing the forelimbs to rest on the workbench. Efficacy of the formulation for skin permeation were analyzed by two assays; toe-spread assay and rota-rod assay. Toe spread and rotarod assay were observed for every 24 hr for 10 days.

[0115] Assay

[0116] Efficacy of the formulation for skin permeation was analyzed by two assay; toe-spread assay and rota-rod assay. Toe spread and rotarod assay were observed for every 24 hr for 10 days. Formulations were applied on left hind limb of the mice and vehicle control (solution C as a control) w as applied to the right hind limb of the mice. Toe-spread data were analyzed using DAS (digital abduction score) as follows: Zero DAS score indicates complete spread of toe and DAS score four indicated complete paralysis. BoNT / A formulations in solution C was applied in both the hind limbs and monitored for 10 days. DAS score was measured as before. For rota rod experiment, each group of trained mice underwent on rotarod and number of rotation mice w ere able to remain on the rotarod w as recorded. Mice were analyzed every day for 10 days for DAS score and number of rotation.

[0117] Result:

[0118] The present formulation was capable to deliver 150 kDa molecule through the skin without any aid of chemical, mechanical or electrical enhancement technique. Moreover, the improved skin permeation is achieved by the present formulation in lower units.

[0119] Example 4:Transport of IgG (~ 150 kDa) through human skinHuman skin permeation studies were conducted using vertical glass Franz diffusion cells (Syncbio Research Pvt. Ltd., Ahmedabad. India). Human IgG was biotinylated using the EZ- Link™ sulfo- manufacturer’s protocol. Human skin (donated by Science Care, Phoenix, AZ, USA) was mounted in the diffusion cells and skin surface measured using a calibrated infrared thermometer. Skin surface temperatures were within 32°C + 1°C. All the experiments were performed in environmentally controlled temperature at 21°C+ 2°C and 50±20% relative humidity to simulate in vivo conditions. Trans-epidermal water loss (TEWL) and skin temperature were monitored, with the skin thickness measured using a calibrated micrometer screw gauge. For each experiment, 30 pL was applied to the donor compartment, and receptor medium (PBS, pH 7.4) was replenished at designated time points (0, 1, 2, 3, 4, 6, 8, 12, 16, 20, 24, 28. 32. and 36 h). Biotinylated human IgG in the receptor compartment was quantified using ELISA with streptavidin-HRP. NHS-LC-Biotinylation Kit (Thermo Scientific, Waltham, MA, USA) according to the manufacturer protocol.Table 3Results:Finite dose human skin permeation studies were conducted over a 36 h period to evaluate the transdermal transport of formulated human IgG. The results are shown in Figure 6. The cumulative amount of IgG passing through the human skin was measured over time. The data clearly indicate the passage of IgG, with the highest transdermal transport observed in the formulation containing 375 pg of IgG. Transport was observed to occur predominantly between2 and 36 h after application. The maximum flux was observed 12 - 24 h, reaching approximately 10 + 5 pg / h.The present formulation was capable of delivering IgG molecule (~ 150 kDa) through the skin without any aid of chemical, mechanical or electrical enhancement technique. Moreover, the improved skin permeation is achieved by the present formulation in lower units.Although the invention has been described in detail in the foregoing for the purpose of illustration, it is to be understood that such detail is solely for that purpose and that variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention except as it may be limited by the claims.

Claims

Claims1. A formulation with improved skin permeability comprising:A. an active pharmaceutical or nutraceutical agent;B. composition B; wherein the composition B comprises: i. lubricant in an amount of 0.5 to 20% by weight based on the total weight of composition B; ii. saponin or its salt thereof in an amount of 50 to 90% by weight based on the total weight of composition B; iii. Cs - C20 fatty acid or a salt thereof; mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof; squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof in an amount of 9.5 to 40 % by weight based on the total weight of composition B.

2. The formulation as claimed in claim 1, wherein the pharmaceutical agent selected from the group consisting of analgesics, antibiotics, anti-infective, antibodies, antiepileptics, antigens from plants, antirheumatics, benzimidazole derivatives, betablocker, cardiovascular drugs, chemotherapeutics, CNS drugs, digitalis glycosides, gastrointestinal drugs, enzymes, hormones, oligonucleotides, peptidhormon proteins, therapeutical bacteria, peptides, proteins, urology drugs, vaccines or mixture thereof.

3. The formulation as claimed in claim 1, wherein the pharmaceutical agent is a protein selected from BoNT / A.

4. The formulation as claimed in claim 1, wherein the pharmaceutical agent is an antibody selected from Immunoglobulin G (IgG).

5. The formulation as claimed in claim 1, wherein the molecular weight of the pharmaceutical agent is in the range of 100 Da to 150 kDa.

6. The formulation as claimed in claim 1, wherein the nutraceutical agent selected from the group consisting of resveratrol from grape products as an antioxidant, soluble dietary fiber products such as psyllium seed husk for reducing hypercholesterolemia, broccoli (sulphane) as a cancer preservative, and soy or clover (isoflavonoids) toimprove arterial health, flavonoids, antioxidants, alpha-linoleic acid from flax seed, beta-carotene from marigold petals or antocyanins from berries or mixtures thereof.

7. The formulation as claimed in claim 1, wherein the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, retinoids, alpha hydroxyl acid, beta hydroxy acid, resveratrol, stem cell, EGF (epidermal growth factors), KGF (keratinocyte growth factors), FGF (fibroblast growth factors), HGH (human growth hormones), niacinamide, aloe vera, allantoin, or a combination thereof.

8. The formulation as claimed in claim 7, wherein the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, or a mixture thereof.

9. The formulation as claimed in claim 8. wherein the lubricant is sodium hyaluronate.

10. The formulation as claimed in claim 1, wherein the lubricant in composition B is present in an amount of 0.5 to 10 % by weight, based on total weight of composition B.

11. The formulation as claimed in claim 1, wherein the saponins comprises of the compounds containing a steroid or triterpenoid aglycone linked to one or more sugar moieties, wherein the saponin in composition B. is present in an amount of 65 to 85 % by weight, based on total weight of composition B.

12. The formulation as claimed in claim 1, wherein the ratio of lubricants to saponins is in the range of 1:20 to 1:50.

13. The formulation as claimed in claim 1, wherein the Cs to C20 fatty acid or a salt thereof, or mono-, di-, tri- glycerides of Cs to C20 fatty acid or a salt thereof is selected from a-linolenic acid, stearidonic acid, eicosapentaenoic acid, cervonic acid, linoleic acid, linolelaidic acid, linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid,mead acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, tricaprylin, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, nonadecylic acid, arachidic acid, or salts thereof, mono-, di-, tri- glycerides of Cs to C20 fatty acid or a salt thereof, or a combination thereof in an amount of 10 to 30 % by weight, preferably 15 to 25 % by weight, each based on total weight of composition B.

14. The formulation as claimed in claim 1, wherein the composition may contain surfactant, emulsifier, preservative, humectants, emollients, stabilizer or a combination thereof.

15. The formulation as claimed in claim 14, wherein the surfactant is selected from sorbitan fatty acid esters such as polyoxyethylene sorbitan and its monolaurate and monooleates (e.g., Tween 20. Tween 60, or Tween 80), sorbitan palmitate, oleate, and stearates (e.g., Span 40, Span 60, Span 65, or Span 80); polyoxyethylene esters; polyethylene glycol fatty acid esters such as Cremophor™; diethylene glycol monolaurate, triethanolamine oleate, ethyl laurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 1 88; and the preservatives may be selected from the group comprising cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, and / or mixtures thereof.

16. The formulation as claimed in claim 14, wherein the preservatives are selected from the group consisting of cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, phenoxyethanol, ethylhexylglycerol, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate calcium propionate alone or mixtures thereof.

17. The formulation as claimed in any one of the claims 1 to 16, wherein the composition is a nanoemulsion.

18. A nanoemulsion with improved skin permeability comprising: i. an aqueous component in an amount of 50% to 97 % by weight, based on total weight of nanoemulsion;ii. a surfactant and / or an emulsifier in an amount of 0.5 % to 15 % by weight, based on total weight of nanoemulsion; iii. an oil component in an amount of 2.5 % to 35 % by weight, based on total weight of nanoemulsion; wherein the aqueous component comprises an active Pharmaceutical or nutraceutical agent, lubricant, saponin or its salt thereof; and wherein the oil component comprises Cs - C20 fatty acid or a salt thereof, mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof, squalene, hydrogenated polyisobutene, microalgal oil, or a mixture thereof.

19. The nanoemulsion according to any one of the embodiments 17 to 18, wherein the nanoemulsion comprises 85 to 97 % by weight of the aqueous component, 2.5 to 10 % by weight of the oil component, and 0.5 to 5 % by weight of the surfactant, based on the total weight of the nanoemulsion.

20. The nanoemulsion as claimed in claim 17, wherein the nanoparticles present in the nanoemulsion have an average particle size distribution of 0.5 to 500 nm, more preferably 0.5 to 100 nm, even more preferably 0.5 to 50 nm, most preferably of 1 to 50 nm, and in particular 1 to 25 nm.

21. The nanoemulsion as claimed in claim 17, wherein the nanoparticles in the nanoemulsion have a zeta potential of 20 to 38.6 mV.

22. The nanoemulsion as claimed in claim 17. wherein the hydrophilic-lipophilic ratio is in the range of 10:0.5 to 40: 1.

23. The nanoemulsion as claimed in claim 17 and 18, wherein the nanoemulsion is an oilin-water-type emulsion.

24. The nanoemulsion as claimed in claim 18, wherein the pharmaceutical agent in the aqueous component of said nanoemulsion is selected from the group consisting of analgesics, antibiotics, anti-infective, antibodies, antiepileptics, antigens from plants, antirheumatics, benzimidazole derivatives, beta-blocker, cardiovascular drugs, chemotherapeutics, CNS drugs, digitalis glycosides, gastrointestinal drugs, enzymes,hormones, oligonucleotides, peptidhormon proteins, therapeutical bacteria, peptides, proteins, urology drugs, vaccines or mixture thereof.

25. The nanoemulsion as claimed in claim 24, wherein the pharmaceutical agent is a protein selected from BoNT / A.

26. The nanoemulsion as claimed in claim 24, wherein the pharmaceutical agent is an antibody selected from Immunoglobulin G (IgG).

27. The nanoemulsion as claimed in claim 18, wherein the nutraceutical agent in the aqueous component of said nanoemulsion is selected from the group consisting of resveratrol from grape products as an antioxidant, soluble dietary fiber products such as psyllium seed husk for reducing hypercholesterolemia, broccoli (sulphane) as a cancer preservative, and soy or clover (isoflavonoids) to improve arterial health, flavonoids, antioxidants, alpha-linoleic acid from flax seed, beta-carotene from marigold petals or antocyanins from berries or mixtures thereof.

28. The nanoemulsion as claimed in claim 18, wherein the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, retinoids, alpha hydroxyl acid, beta hydroxy acid, resveratrol, stem cell, EGF (epidermal growth factors), KGF (keratinocyte grow th factors), FGF (fibroblast growth factors), HGH (human growth hormones), niacinamide, aloe vera, allantoin, or a combination thereof.

29. The nanoemulsion as claimed in claim 28, wherein the lubricant is selected from hyaluronic acid or its salt, hydrolyzed hyaluronic acid or its salt, acetylated hyaluronic acid or its salt, hydrolyzed acetylated hyaluronic acid or its salt, or a mixture thereof.

30. The nanoemulsion as claimed in claim 29, wherein the lubricant is sodium hyaluronate.

31. The nanoemulsion as claimed in claim 18, wherein the saponins comprises of the compounds containing a steroid or triterpenoid aglycone linked to one or more sugar moieties.

32. The nanoemulsion as claimed in claim 18, wherein the surfactant is selected from sorbitan fatty acid esters such as polyoxyethylene sorbitan and its monolaurate and monooleates (e.g., Tween 20. Tween 60, or Tween 80), sorbitan palmitate, oleate, and stearates (e.g., Span 40, Span 60, Span 65, or Span 80); polyoxyethylene esters; polyethylene glycol fatty acid esters such as Cremophor™; diethylene glycol monolaurate, triethanolamine oleate, ethyl laurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 1 88; and the preservatives may be selected from the group comprising cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, and / or mixtures thereof.

33. The nanoemulsion as claimed in claim 18. wherein the emulsifier is selected from is selected from polyglycosides, alcohols, sugar and sugar derivatives, polyethers, amines, polyethylene derivatives, alky l sulfates (e.g. sodium dodecyl sulfate), alkyl ether sulfates, dioctyd sodium sulfosuccinate, polysorbates (e.g. polyoxyethylene (20) sorbitan monooleate), nonylphenol ethoxylates (nonoxynol-9), alkyl polyglycosides, decyl glucoside, decyl polyglucose, lauryl glucoside, octyl glucoside, N-octyl beta- D-thioglucopyranoside, cetostearyl alcohol, cetyl alcohol, stearyl alcohol, polyoxyethylene cetostearyl alcohol, cetylstearyl alcohol, oleyl alcohol, polyglyceryl-6-dioleate, glyceryl stearate citrate, polyglyceryl-3 caprate, polyglyceryl-3 diisostearate, glyceryl isostearate, polyglyceryl-4-isostearate, glyceryl monolinoleate, dicaprylyl carbonate, alcohol polyglycol ether, polyethylenglycolether of cetearyl alcohols (n=20), polyethylene glycol-6 stearate, glycol stearate, polyethylene gly col-32 stearate, polyethylene gly col-20 stearate, fatty alcohol polyglycol ether, polyethylene glycol-4 laurate, polyethylene glycol isocetyl ether (n=20). poly ethylenegly col-32 (Mw 1500 g / mol) mono- and diesters of lauric acid (C12), nonaethylene glycol, polyethylene glycol nonylphenyl ether, octaethylene glycol monododecyl ether, pentaethylene glycol monododecyl ether, polyethylene glycol macrocetyl ether, polyethylene glycol esters of palmitic (Cie) or stearic (Cis) or caprylic acids, polyoxyethylene fatty ether derived from stearyl alcohols like BRIJ S2, polyoxyethylene oxypropylene stearate, macrogol stearyl ether (20),diethylaminoethyl stearate, polyethylene glycol stearate, sucrose distearate, sucrose tristearate, sorbitan monostearate, sorbitan tristearate, mannide monooleate, octaglycerol monooleate, sorbitan dioleate, polyricinoleate, polysorbate like polysorbate 20 and Polyoxyethylene (20) sorbitan monooleate (polysorbate 80), sorbitan, sorbitan monolaurate, sucrose cocoate, glycereth-2 cocoate, ethylhexyl cocoate, polypropylene glycol -3 benzyl ether myristate, sodium myristate, gold sodium thiomalate, polyethylene glycol 8 laurate, polyethylene-4 dilaurate, from a- Hexadecyl-cj-hydroxypoly(oxyethylene), cocamide diethanolamine, N-(2- hydroxyethyl)dodecanamide, octylphenoxy polyethoxyethanol, maltoside, 2,3- Dihydroxypropyl dodecanoate, 3-[(3R,6R,9R,12R.15S,22S,25S.30aS)-6,9, 15,22- Tetrakis(2-amino-2-oxoethyl)-3-(4-hydroxybenzyl)-12-(hydroxyl methyl)-18-(l 1- methyltridecyl)-! ,4,7,10,13, 16,20,23,26- nona oxo triaconta hydropyrrolo[l,2-g][l ,4,7, 10,13, 16, 19,22,25]nonaazacyclooctacosin-25-yl jpropenamide, 2- {2-[2-(2- {2- [2-(2-{2-[2-(4-nonylphenoxy)ethoxy] ethoxy} ethoxy) ethoxy] ethoxy} ethoxy)ethoxy] ethoxy} ethanol, oxy poly ethoxy dodecane, poloxamers like poloxamer 188 (Pluronic F-68) and poloxamer407, propylene glycol monocaprylate type I (Capryol PGMC), polyethoxylated tallow amine, polyglycerol, polyoxyl 40 hydrogenated castor oil, surfactin, 2-[4-(2,4,4-trimethylpentan-2- yl)phenoxy]ethanol, carbomer, sodium carbomer. carboxymethylcellulose calcium, carrageenan, cholesterol, deoxycholic acid, phospholipids like egg phospholipids, gellan gum, lanolin, capric acid, waxes like Polawax NF, Polawax A31 or Ceral PW, ester gum, dea-cetyl phosphate, soya lecithin, sphingomyelins, sodium phosphate, sodium lauroyl lactylate, lanolin, Oxirane methyl- polymer with oxirane monobutyl ether. 1,2-dierucoylphosphatidylcholine, dimethicone end-blocked with an average of 14 moles of propylene oxide, laurylmethicone copolyol, lauroglycol 90, white mineral oil like Amphocerine KS, dispersion of acrylamide / sodium acryloyldimethyl taurate copolymer in isohexadecane, and sodium polyacrylate or mixtures thereof. Preferred are macrogol stearyl ether (20) and polysorbate 80, or a mixture thereof.

34. The nanoemulsion as claimed in claim 18, wherein the Cs to C20 fatty acid or a salt thereof, or mono-, di-, tri- glycerides of Cs to C20 fatty acid or a salt thereof is selected from a-linolenic acid, stearidonic acid, eicosapentaenoic acid, cervonic acid, linoleic acid, linolelaidic acid, linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid,mead acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, tricaprylin, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, nonadecylic acid, arachidic acid, or salts thereof, mono-, di-, tri- glycerides of C5 to C20 fatty acid or a salt thereof, or a combination thereof.

35. The nanoemulsion as claimed in claim 18, wherein the nanoemulsion may contain preservative, humectants, emollients, stabilizer or a combination thereof.

36. The nanoemulsion as claimed in claim 18, wherein the amount of fatty acids present in the oil component is in the range of 9.5 to 40% by weight, based on total weight of the nanoemulsion, more preferably 10 to 35% by weight, even more preferably 10 to 30 % by weight, most preferably 10 to 55% by weight, and in particular preferably 15 to 25 % by weight.

37. The nanoemulsion as claimed in claim 18. wherein the ratio of lubricants to saponins in the aqueous phase is in the range of 1:20 to 1:50.

38. The nanoemulsion as claimed in claim 18. wherein surfactant and / or emulsifier is in an amount of 0.5 to 50% by weight of the total composition of the nanoemulsion.

39. The nanoemulsion as claimed in claim 18, wherein the aqueous component in the nano emulsion may further comprise a co-solvent, preferably an organic solvent, more preferably water miscible organic solvent in an amount of 65 to 95 % by weight of solvent based on total weight of the nanoemulsion.

40. The nanoemulsion as claimed in claim 18, wherein the nanoemulsion is stable at a pH in the range of 5.5 to 8.0.

41. A process for preparation of the formulation or the nanoemulsion as claimed in any one of the claims 1 to 40 comprising; i. preparing an aqueous solution by mixing lubricant, surfactant and / or emulsifier and saponin in water by continuous stirring; ii. preparing an oil phase; andiii. mixing oil phase into aqueous solution under continuous stirring to obtain an emulsion; wherein the active pharmaceutical or nutraceutical agent dissolved in a solvent or buffer is mixed in aqueous solution or oil phase or mixed to nanoemulsion of step (iii) obtain the desired nanoemulsion.

42. The process as claimed in claim 41, wherein the aqueous phase comprising lubricant, surfactant and / or emulsifier and saponin may be lyophilized to obtain a powder which was then mixed in the oil phase and the active ingredients to obtain the product.

43. The formulation / nanoemulsion as claimed in any one of the claims 1 to 42 for use in the treatment, prevention, inhibition, delaying onset of, or causing regression of one or more neuro-muscular diseases and conditions or treatment of wrinkle or used in cosmetic treatments, diseases or conditions which include neuronal regeneration / sprouting. disease involving muscle movement, various wounds, scars and gastrointestinal symptoms.

44. The method of treating, preventing, inhibiting, delaying onset of, or causing regression of one or more neuro-muscular diseases and conditions or treatment of wrinkle or used in cosmetic treatment, diseases or conditions which include neuronal regeneration / sprouting, disease involving muscle movement, various wounds, scars and gastrointestinal symptoms comprising applying said formulation / nanoemulsion as claimed in any one of the claims 1 to 42.

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