DESC:Field of Invention:
The present invention relates to compositions comprising plant extract and process of preparation thereof. Particularly, the invention relates to the composition comprising Ashwagandha extract containing 0.5-10% of Withanolide glycoside encapsulated in a natural protective coat exhibiting enhanced bioactivity and bioavailability.

Background of the invention:
Ashwagandha or Withania Somnifera is an adaptogenic herb, which means that it helps the body to adapt to stress. Ashwagandha (Withania somnifera) belongs to the Solanaceae family and commonly also known as “Indian Winter cherry” or “Indian Ginseng”. Ashwagandha finds an important place in Ayurveda. It has been shown to be effective in reducing anxiety and stress, and it can also improve stamina. The bioactivity of ashwagandha is due to its active ingredients, which include withanolides and steroidal lactones. These compounds have been shown to have a variety of effects on the body, such as reducing cortisol levels. Cortisol is a stress hormone the increased levels of which have negative effects on the body, like increasing anxiety and fatigue. Ashwagandha reduces the cortisol levels, which can help to improve mood and reduce stress. Ashwagandha also increases GABA levels, thus it helps to reduce anxiety and promote relaxation. Ashwagandha improves cognitive function in people with anxiety and stress. Ashwagandha improves memory, focus, and concentration. Immunity of an individual is of utmost importance to fight illness in a stressful life, Ashwagandha also boosts the immune system.

Ashwagandha is commonly available as a churna, a fine sieved powder that can be mixed with water, ghee (clarified butter) or honey.

WO2017/068600 discloses compositions and processes of Ashwagandha extract enriched with Withanolide glycosides and saponins after removing alkaloids, withanolide aglycones and oligosaccharides.

WO2023/021434 describes a modified release formulation comprising 2 to 10% withanolides, obtained from root powder of Withania somnifera. The formulation is comprised of at least one pH independent release controlling agent or the combination thereof. The invention also provides a process for preparation of modified release composition; wherein Ashwagandha root or whole plant extract powder enriched with withanolides is uniformly dispersed and embedded throughout the matrix of at least one pH independent release controlling agent. The granules can be conveniently formulated in suitable compressible dosage forms, filled in sachets or capsules for oral administration. The formulation is designed to have less than 20% release in Stomach and Not less than 90% release in Intestine over a period of 8 to 12 hours.

Further there are many prior art documents available pertaining to compositions comprising Ashwagandha and/or extract of Ashwagandha, encapsulating the said composition by synthetic polymeric coating material.

However synthetic polymeric coating of Ashwagandha would face certain drawbacks such as reduced absorption, increased side effects, environmental impact and the added cost. The synthetic polymer coats can reduce the amount of active ingredients that are absorbed into the bloodstream. Synthetic polymer coats can sometimes irritate the stomach lining, which can lead to side effects such as nausea and vomiting. The Synthetic polymers are not biodegradable, thus would add to the environment pollution.

Synthetic polymer coats do not offer the same level of protection to ashwagandha. They could be degraded in the stomach, which may reduce the amount of active ingredients that are absorbed.

Ashwagandha gets hydrolysed in stomach acid and gets degraded thus lowering its bioactivity. Due to this a higher dose is required to have the desired bioactivity.
Considering the drawbacks of the prior art literature and the problems of the existing compositions comprising Ashwagandha the inventors of the present invention have formulated compositions comprising Ashwagandha extract which are protected in the stomach environment and more effectively absorbed in the small intestine by encapsulating in the protective coat of natural origin..

Objects of the invention
It is the primary objective of the present invention to provide compositions comprising plant extract encapsulated in the natural protective coat that exhibits enhanced bioactivity and bioavailability and to the process for preparation thereof.

Another object of the present invention is to provide composition comprising Ashwagandha extract encapsulated in the natural protective coat with enhanced bioactivity and sustained release of the active Withanolide glycoside.

Summary of the Invention:
In accordance with the above objects, the present invention provides a composition comprising plant extracts with enhanced bioactivity and a process of preparation thereof.

The plant extracts comprise of Ashwagandha, curcuminoid, quercetin, green tea extracts, sylemarin extract or Boswellia extract.

In a preferred aspect, the present invention provides a composition comprising Ashwagandha extracts with enhanced bioactivity and to the process for preparation thereof.

In an aspect, the Ashwagandha extract present in the composition is protected by a natural protective coat comprising of a lipid, polysaccharide and protein which thereby helps in creating acid resistant coating on Ashwagandha thus protecting it from stomach acid degradation and increased absorption in intestine thereby reducing the dose of active. The protective coat further provides a sustained release of the active Withanolide Glycoside and maintains the level of the active in the blood over an extended time period.

Accordingly, the present invention provides a composition comprising Ashwagandha extract containing 0.5-10% w/w of Withanolide glycoside in a natural protective coat selected from lipids, polysaccharides and protein.

In another aspect, the Ashwagandha extract comprises the water extract of Ashwagandha root or other plant parts containing 0.5 % to 3.5 % w/w of Withanolide glycoside as active and the hydroalcoholic extract of Ashwagandha powder containing 0.5 % to 15 % w/w of Withanolide glycoside as active.

In an aspect, the Hydroalcoholic extract of Ashwagandha and the water extract of Ashwagandha are in the ratio of 5:95 to 95:5.

In yet another aspect, the lipids are selected from Medium- chain triglyceride (MCT) oil, Sunflower oil, Sea buckthorn oil, Almond oil, Olive oil, Moringa Oil, Withania Oil, Fenugreek Oil, Flaxseed oil, Moringa oil, Cumin oil, Grape seed oil, Evening prime rose oil, or mixtures thereof, and other oils with specification of stearic acid in the concentration of 0.1% to 60% w/w, Oleic acid in the concentration of 0.1 % to 90% w/w, Palmitic acid in the concentration of 0.1 % to 60% w/w, Linoleic acid in the concentration of 0.1% 60% w/w.

In another aspect, the polysaccharides are selected from natural starch such as Rice starch, Potato starch, Tapioca starch, Corn starch; Modified starch selected from Pregelatinized starch, Maltodextrin or mixtures thereof in a concentration ranging from 1% to 50% w/w.

In an aspect, the plant protein is selected from Rice protein, Pumpkin seed protein, Almond protein, Pea protein, oat protein, hemp seed protein, sunflower protein, mung protein, fenugreek protein or mixtures thereof in a concentration ranging from 1% to 75% w/w.

In another aspect, the composition of the present invention may comprise natural hydrocolloids, cellulose gum and emulsifiers. The natural hydrocolloids comprises plant pectin, Gum acacia, Guar gum or mixtures thereof in a concentration ranging from 1% to 50% w/w. The emulsifiers are selected from sunflower lecithin, soya lecithin, Datem and Plant Saponins or mixtures thereof in a concentration range of 0.5 % to 5% w/w.

The extract of Ashwagandha may be prepared from plant parts such as roots, stems, leaves, barks, seeds of Ashwagandha.

In an aspect, the present invention provides a process for preparing the composition of encapsulated Ashwagandha extract in a natural protective coat comprising;
i. Preparing the protective coat by adding lipids in water and mixing followed by addition of emulsifiers, polysaccharides, protein and mixing;
ii. Preparing the soluble matrix of Ashwagandha by mixing the hydroalcoholic extract of Ashwagandha powder with the water extract of Ashwagandha root or other plant parts with continuous stirring; and
iii. Mixing the protective coat of step (i) with the soluble matrix of Ashwagandha of step (ii) followed by drying to obtain the desired product.
OR
i. Homogenising Ashwagandha hydroalcoholic extract with water extract of Ashwagandha in liquid form for a specified time interval and further adding the components of the protective coat which comprise of oil, Lecithin, Polysaccharide and protein to form a slurry; and
ii. Drying the slurry.

The present invention provides the composition of Ashwagandha extract that exhibits enhanced bioactivity and bioavailablity of the active Withanolide glycoside.

Description of the Figures
Figure 1 depicts Gastric acid-resistant profile of the developed formulation (E18 and E38) against Ashwagandha extract in 1.2 pH. The gastric acid resistance property of formulation compared to the extract was checked i.e. the extract released from the formulation was observed very less (light color) compared to the extract standalone (dark color).

Figure 2 depicts the Release profile of Ashwagandha in different formulation and extract in 1.2 pH. -Unformulated Ashwagandha extract (dark blue), Formulation E34(light blue), Formulation E26(orange), Formulation E31(yellow) and Formulation E30 (grey).

Figure 3 depicts Pictorial illustration of wavelength maxima of Ashwagandha Extract. The optimal wavelength for ashwagandha extract was found to be 227 nm.

Figure 4 depicts Calibration curve of Ashwagandha extract at 227nm.

Figure 5 depicts release profile of Ashwagandha in different formulation and extract in 1.2 pH followed by 6.8 pH. Ashwagandha extract(dark blue), Formulation E18(orange), Formulation E33(grey).

Figure 6 depicts high-performance thin layer chromatography (HPTLC) finger printing of Ashwagandha extracts and formulations at UV 366 (A), UV 254 (B) prior to derivatization, and at UV 366 (C), White light (D) after derivatization.

Detailed Description of the invention:
Geographical origin:
Biological material
Geographical origin Source
Ashwagandha
(Withania somnifera)
It is an evergreen shrub that grows in Asia and Africa. Ashwagandha is a pantropic native medicinal plant growing all over north-western and central India. India
Sunflower Belongs to the family Asteraceae. The plant was first domesticated in the America. India
Coconut Is a member of the palm tree family (Arecaceae) and the only living species of the genus Cocos. Native to southeast Asia. India
Sea buckthorn It is a deciduous shrub in the family Elaeagnaceae. It extends from the Atlantic coasts of Europe across to northwestern Mongolia, northwestern China and Northern Pakistan. India
Almond It is a species of tree from the genus Prunus. Its origins were in Central Asia between Iran, Turkmenistan, Tajikistan, Kurdistan, Afghanistan, and Iraq,[26][27] or in an eastern Asian subregion between Mongolia and Uzbekistan. India
Soy Is a species of legume native to East Asia India
Evening -prime Rose Is a species of flowering plant in the family Onagraceae, native to eastern and central North America, from Newfoundland west to Alberta, southeast to Florida, and southwest to Texas. China
Olive It is a species of small tree or shrub in the family Oleaceae, found traditionally in the Mediterranean Basin, with wild subspecies found further afield in Africa and western Asia. USA
Moringa It belongs to the plant family Moringaceae. It occur in tropical and subtropical regions of Africa and Asia. India
Withania It is a genus of flowering plants in the nightshade family, Solanaceae, is native to parts of North Africa, western Asia, south Asia, southern Europe, the Mediterranean, and the Canary Islands India
Fenugreek It is an annual plant in the family Fabaceae. It originated in the Near East. India
Flaxseed It belongs to the family Linaceae. Originated in present day Georgia. India
Grape Is a fruit, botanically a berry, of the deciduous woody vines of the flowering plant genus Vitis. Originated in Middle east. Chile
Rice A cereal grain originated in Asia and Africa. India
Potato Is a starchy root vegetable native to America. India
Tapioca Is a starch extracted from the tubers of the cassava plant (Manihot esculenta, also known as manioc), a species native to the North and Northeast regions of Brazil. India
Corn Is a tall stout grass that produces cereal grain. It was domesticated by indigenous peoples in southern Mexico. India
Pumpkin Is a cultivated winter squash in the genus Cucurbita. Originated in New England in North America. India
Pea Is a pulse, vegetable or fodder crop, Pisum sativum. Originated in the Mediterranean Basin and the Near East. India
Oat Is a tall stout grass, a member of the family Poaceae. Originated in western Europe. India
Hemp Is a plant in the botanical class of Cannabis sativa cultivars, originated in china India
Mung Green gram (Vigna radiata) is a plant species in the legume family. Cultivated in East, Southeast and South Asia. India
Apple Belongs to the Malus spp. The tree originated in Central Asia. India
Citrus Belongs to the family Rutaceae. Is native to South Asia, East Asia, Southeast Asia, Melanesia, and Australia. India
Tamarind Is a leguminous tree bearing edible fruit that is indigenous to tropical Africa and naturalized in Asia. India
Gum acacia Is a gum of Acacia tree, originated in Africa. India
Guar gum Extracted from guar beans. Principally grown in India, Pakistan, the United States, Australia and Africa. India

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. As used herein, the following terms and phrases shall have the meaning set forth below.

Unless specified otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art, to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described.

Unless stated to the contrary, any of the words “contains”, “containing”, "including," "includes," "comprising," and "comprises" mean "including without limitation" and shall not be construed to limit any general statement that it follows to the specific or similar items or matters immediately following it. Embodiments of the invention are not mutually exclusive, but may be implemented in various combinations. The described embodiments of the invention and the disclosed examples are given for the purpose of illustration rather than limitation of the invention.

Further, words like “a”, “an”, “at least” and “the” should be construed to not only cover singular quantities but also plural quantities of the elements immediately following them.

The term “Natural protective coat” used in the entire specification refers to the protective coat comprising plant based protective coats.

The present invention relates to the compositions with enhanced bioactivity and bioavailability comprising the plant extracts encapsulated in the natural protective coat.

The plant extracts are selected from Ashwagandha, curcuminoid, quercetin, green tea extracts, sylemarin extract or Boswellia extract.

In a preferred embodiment, the composition of the present invention comprises the Ashwagandha extract.

The water extract of Ashwagandha and the Hydroalcoholic extract of Ashwagandha used for the present invention were procured from Indian sources. These extracts of Ashwagandha comprise the active Withanolide Glycoside in defined percentages.

The water extract of Ashwagandha would mean an extract, water extract of Ashwagandha root or plant with total solid content in the range of 5% to 95%w/w with Withanolides 0.5% to 3.5% w/w.

The Hydroalcoholic extract of Ashwagandha would mean an extract, water/methanol or water /ethanol extract of Ashwagandha root or plant part with Withanolides 0.5% to 15% w/w.

Compositions comprising Ashwagandha extract are been provided which exhibit increased Bioactivity and with reduced doses.

The present invention provides compositions which exhibit sustained release effect over an extended time period.

The invention also provides a process for preparation of the compositions comprising Ashwagandha extract which exhibit increased Bioactivity and with reduced doses.

In an embodiment, an Ashwagandha extract composition for enhancing the bioactivity of Ashwagandha extracts comprises of water extract of Ashwagandha root or plant parts and hydroalcoholic extract of Ashwagandha root or plant parts along with the protective coating.

In an embodiment, the water extract of Ashwagandha root or plant parts contains Total Dissolved Solids (TDS) in the range of 5% to 95% w/w.

In an embodiment, the water extract of Ashwagandha root or plant parts comprises of 0.5 % to 3.5 % w/w of Withanolide glycoside.

In another embodiment, the Hydroalcoholic extract comprises of 0.5 % to 15 % w/w of Withanolide glycoside.

In a further embodiment, the Hydroalcoholic extract of Ashwagandha and the water extract of Ashwagandha are mixed in the ratio of 5:95 to 95:5.

The extract of Ashwagandha may be prepared from plant parts such as roots, stems, leaves, barks, seeds of Ashwagandha.

In yet another embodiment, the protective coating of Ashwagandha and Ashwagandha extract comprises of lipid, polysaccharides and protein.

In an embodiment, the present invention relates to a composition comprising the encapsulated Ashwagandha extract containing 0.5-10% w/w of Withanolide glycoside in a protective coat selected from lipids, polysaccharides, protein or mixtures thereof.

In yet further embodiment, the compositions of the present invention provides a sustained release effect in which the active released in a sustained manner over a time period.

The Oils that provide the source of lipid is selected from a natural source and comprises Medium- chain triglyceride (MCT) oil, Sunflower oil, Sea buckthorn oil, Almond oil, Olive oil, Moringa Oil, Withania Oil, Fenugreek Oil, Flaxseed oil, Moringa oil, Cumin oil, Grape seed oil, Evening prime rose oil or mixtures thereof.

The lipid source may be oils specified with Stearic acid in a concentration ranging from 0.1% to 60% w/w, Oleic acid in a concentration ranging from of 0.1 % to 90% w/w, Palmitic acid in a concentration ranging from 0.1 % to 60% w/w, Caprylic acid in a concentration ranging from 0.1 % to 60% w/w and Linoleic acid in a concentration ranging from 0.1% 60% w/w or mixtures thereof.

The Plant protein are in a concentration ranging from 1% to 75% selected from Rice protein, Pumpkin seed protein, Almond protein, Pea protein, oat protein, hemp seed protein, sunflower protein, mung protein, fenugreek protein or mixtures thereof.

The Natural starch which serves as a source of polysaccharide are present in a concentration ranging from 1% to 50% w/w and are selected from Rice starch, Potato starch, Tapioca starch, Corn starch; Modified starch selected from Pregelatinized starch, Maltodextrin and the natural gum like; xanthan gum, gum acacia, Cellulose gum selected from methyl cellulose; Hydroxypropyl Methyl Cellulose, ethyl cellulose or mixtures thereof.

The composition of the present invention may further comprise natural hydrocolloids, cellulose gum, and emulsifier in defined range of concentration.

The natural hydrocolloids comprise plant pectin, Gum acacia, Guar gum or mixtures thereof in a concentration ranging from 1% to 50% w/w. The pectin is sourced from Apple, citrus, or Tamarind.

The cellulose gum is selected from methyl cellulose, Hydroxypropyl Methyl Cellulose, ethyl cellulose or mixtures thereof.

The emulsifiers are selected from sunflower lecithin, soya lecithin, Datem (diacetyl tartaric acid ester of mono- and diglycerides) and Plant Saponins or mixtures thereof in a concentration range of 0.5 % to 5% w/w.

In another embodiment of the invention, the present invention discloses the process for preparation of Ashwagandha extract composition encapsulated by a protective natural coat wherein the water extract of Ashwagandha root or other plant parts with Total dissolved Solids (TDS) present in a concentration of 5% to 95% w/w and Hydroalcoholic extract of Ashwagandha powder present at a concentration of 0.5% to 15% w/w Withanolide glycoside.

In yet another embodiment of the invention, the invention discloses the process for preparation of an Ashwagandha extract composition encapsulated by a protective natural coat wherein the water extract of Ashwagandha having high amount of the polysaccharide helps in entrapment of oil soluble Withanolide.

The water extract of Ashwagandha root or plant contains a significant amount of water-soluble polysaccharides. These polysaccharides possess the ability to form hydrocolloids, which effectively stabilize emulsions and create a gel-like matrix in aqueous solutions. This property is particularly advantageous for the incorporation of oil-soluble withanolides into water-based systems.
In an embodiment, the polysaccharides in the water extract are not more than 35% w/w.
Yet another embodiment the invention provides a single oral administration of an Ashwagandha extract composition in dosage range between 250mg to 300mg per day.

In an embodiment of the invention, the present invention discloses protective coating composition for Ashwagandha extract in powder form wherein lipid, polysaccharides and protein coating material help in creating acid resistance coating on the Ashwagandha and helps in reduction of degradation of actives in stomach pH.

In another embodiment, the invention relates to a process for preparation of the composition comprising Ashwagandha extract encapsulated by a natural protective coat comprising the steps of:
i. Preparing the protective coat by adding lipids in water and mixing followed by addition of emulsifiers, polysaccharides, protein and mixing;
ii. Preparing the soluble matrix of Ashwagandha by mixing the hydroalcoholic extract of Ashwagandha powder with the water extract of Ashwagandha root or plant part with continuous stirring; and
iii. Mixing the protective coat of step (i) with the soluble matrix of Ashwagandha of step (ii) followed by drying to obtain the desired product.
OR
i. Homogenising Ashwagandha hydroalcoholic extract with water extract of Ashwagandha in liquid form for a specified time interval and further adding the components of the protective coat which comprise of oil, Lecithin, Polysaccharide and protein to form a slurry; and
ii. Drying the slurry.

Accordingly, the invention discloses the process for preparation of Ashwagandha extract composition having a protective coat wherein the process for preparation of soluble matrix of Ashwagandha extract comprises the step of mixing hydroalcoholic extract of Ashwagandha powder comprising 0.5% to 15% w/w Withanolide glycoside with water extract of Ashwagandha root or other plant parts comprising 0.5 % to 3.5 % w/w of Withanolide glycoside and with Total Dissolved Solids (TDS) of 5% to 95% w/w in 5: 95 to 95:5 ratio with continuous stirring to make slurry.

In another embodiment, the invention discloses a process for preparation of an Ashwagandha extract composition encapsulated by a protective natural coat, wherein the process for preparation of a composition comprising Ashwagandha extract comprises the step of mixing the protective blend with soluble matrix of Ashwagandha extract for a time interval at a defined temperature. The mixture is dried using tray dryer or vacuum drier or Fluid bed dryer or the mixture will be spray dried using a spray drier to obtain the final powder product.

In yet another embodiment, the present invention discloses a process for preparation of composition comprising the Ashwagandha extract wherein the process involves homogenising Ashwagandha hydroalcoholic extract with water extract of Ashwagandha in liquid form for a specified time interval and further adding the components of the protective coat which comprise of oil, Lecithin, Polysaccharide and protein to form a slurry and thereafter drying the slurry either by tray drying, Vacuum drying or Spray drying.

In an embodiment, the composition of the present invention may be formulated in the form of capsule, tablets, granules, sachet, powder, paste, infusion, ampoule, solution, suspension, emulsion, pills and cream.

In an embodiment, the present invention discloses the use of the composition comprising the Ashwagandha extract containing 0.5-10% of Withanolide glycoside as active encapsulated in a natural protective coat for sustained release.

In yet another embodiment, the present invention discloses the composition comprising the Ashwagandha extract containing 0.5-10% w/w of Withanolide glycoside as active encapsulated in a natural protective coat with enhanced bioactivity and bioavailability.

In another embodiment, the present invention discloses a method of enhancing bioactivity of the present composition comprising administering said coated composition of the extract of Ashwagandha.

In an embodiment, the present invention discloses a method of enhancing bioavailability of Withanolide glycosides from an extract of Ashwagandha, comprising administering the encapsulated composition of Ashwagandha extract to a subject in need thereof.

In another embodiment, the present invention relates to a method of sustained release of the Ashwagandha extract in gastrointestinal medium without degradation comprising administering said composition of the Ashwagandha extract, whereby the active Withanolide glycoside in the extract of Ashwagandha is released from the protective coat at the pH of the gastrointestinal medium.

Yet another embodiment of the present invention provides an Ashwagandha extract composition that does not degrade at acidic pH and releases the active content in intestinal pH thereby enhancing the bioactivity.

Further details of the present invention will be apparent from the examples presented below. Examples presented are purely illustrative and should not be construed as limiting the scope of the invention in any manner.

Example 1: Composition
Example No. E1 E2 E3 E4 E5
Ingredients % w/w % w/w % w/w % w/w % w/w
Sunflower Lecithin 5 5 5 5 5
MCT Oil 5 5 5 5 5
Gum Acacia 12 12 12 12 12
Apple Pectin 8 8 8 8 8
Rice Protein 20 - - - -
Pea Protein - 20 - - -
Mung Protein - - 20 - -
Sunflower protein - - - 20 -
Pumpkin protein - - - - 20
Hydroalcoholic extract of Ashwagandha root or other plant parts. 5 to 50 5 to 50 5 to 50 5 to 50 5 to 50
Ashwagandha water extract with total solid content 5 to 95% 5-45 5-45 5-45 5-45 5-45

Example No. E6 E7 E8 E9 E10
Ingredients % w/w % w/w % w/w % w/w % w/w
Sunflower Lecithin - - 5 5 5
Datem 5 5 - - -
MCT Oil 5 5 - - -
Seabuckthorn oil - - 5 - -
Olive oil - - - - 5
Almond oil - - - 5 -
Sunflower oil - 5 - - -
Gum Acacia 12 12 12 12 12
Apple Pectin 8 8 8 8 8
Sunflower protein 20 20 20 20 20
Hydroalcoholic extract of Ashwagandha root or otherplant parts 5 to 50 5 to 50 5 to 50 5 to 50 5 to 50
Ashwagandha water extract l) with total solid content 5 to 95% 5-45 5-45 5-45 5-45 5-45

Example No. E11 E12 E13 E14 E15 E16
Ingredients % w/w % w/w % w/w % w/w % w/w % w/w
Sunflower lecithin 5 4 4 3.5 5 4
MCT oil 7 7 7 7 7 7
Gum acacia 15 8 5 8 5 8
Tamrind Pectin 3 10 10 3 3 10
Pea Protein 20 20 20 20 20 20
Potato starch - - - - - 5
Xanthan gum - - 5 3 2 -
Hydroalcoholic extract of Ashwagandha root or other plant parts 45 5 to 50 5 to 50 5 to 50 5 to 50 5 to 50
Ashwagandha water extract with total solid content 5 to 95% 5 5-45 5-45 5-45 5-45 5-45

Example No. E17 E18 E19 E20 E21
Ingredients % w/w % w/w % w/w % w/w % w/w
Sunflower lecithin 5 5 4 3.5 5
Ashwagandha oil 7 7 7 7 7
Gum acacia 15 5 5 8 5
Apple Pectin 3 8 - - -
Rice Protein 20 - - - -
Pea Protein - 20 20 20 20
Hydroxy propyl methyl cellulose - - 5 3 2
Hydroalcoholic extract of Ashwagandha root or other plant parts 45 45 45 5 to 50 51.5
Ashwagandha water extract with total solid content 5 to 95% 5 5 5 5-45 5-45

Example No. E22 E23 E24 E25 E26 E27
Ingredients % w/w % w/w % w/w % w/w % w/w % w/w
Sunflower lecithin 5 4 4 3.5 3.5 3.5
MCT oil 7 7 7 7 7 7
Gum acacia 15 8 5 8 5 5
Xanthan gum - - 2 5 7 7
Tamrind Pectin 3 10 10 3 3 3
Sunflower Protein - - - - - 20
Pea Protein 20 20 20 20 20 -
Hydroxy propyl methyl cellulose - - - - - -
Hydroalcoholic extract of Ashwagandha root or other plant parts 45 49 45 45 45 45
Ashwagandha water extract with total solid content 5 to 95% 5 1 5 5 5 5

Example No. E28 E29 E30 E31 E32 E33 E34
Ingredients % % % % % % %
Sunflower lecithin 5 5 5 5 5 5 3.5
MCT oil 7 7 8 7 7 7 7
Gum acacia 5 5 5 5 6 5 8
Xanthan gum 2 2 3 2 2 2 -
Tamrind Pectin 8 8 9 8 8 8 3
Sunflower Protein - - - - - - -
Pea Protein 20 20 20 20 20 20 20
Hydroxy propyl methyl cellulose - - - - - - 5
Hydroalcoholic extract of Ashwagandha root or other plant parts 45 45 50 - 45 48 50
Ashwagandha water extract with total solid content 5 to 95% 5 5 - 53 5 2

Example No. E35 E36 E37 E38 E39
Ingredients % % % % %
Sunflower lecithin 5 5 5 5 5
MCT oil 7 7 7 7 7
Gum acacia 5 5 5 5 5
Tamrind Pectin 8 8 8 8 8
Sunflower Protein 20 20 20 20 20
Xanthan Gum 2 2 2 2 2
Curcuminoid extract 53 - - - -
Boswellia extract - 53 - - -
Sylemarin extract - - 53 - -
Green tea extract 53
Quercetin extract 53

Example 2: General process for preparation of the present composition
Step 1: Preparation of the Protective Coat
1. Added lipid from natural source (MCT oil, Sunflower oil, Sea buckthorn oil, Almond oil, Olive oil, Moringa Oil, Hemp seed oil, Evening prime rose oil,other oils with Specification of stearic acid NLT 0.1% to 60%, Oleic acid 0.1 % to 90%, Palmitic acid with 0.1 % to 60%, Linoleic acid with 0.1% 60%) in 50ml of water and mixed for 5-30 minutes;
2. Added emulsifier (like sunflower lecithin, soya lecithin, Datem Plant Saponins eg quillaia extract) to above mixture;
3. Added natural hydrocolloid (like Pectin from Apple, citrus, Tamarind, Gum acacia, xanthan gum) to above mixture and mixed for 5-30 minutes;
4. Added natural starch (Potato starch, Tapioca starch, Corn starch) or Modified starch Pregelatinized starch or Cellulose gum, methyl cellulose to the above mixture and mixed for 5-45 minutes;
5. Added plant protein (like rice protein, Pumpkin seed protein, Almond protein, Pea protein, hemp seed protein, potato protein, Guar protein) to the above mix and further mixed for 5-45 minutes.
Step 2: Preparation of soluble matrix of Ashwagandha containing 0.5% to 10% of Withanolide
1. Prepared Water extract of Ashwagandha root or whole plant with total dissolved solids (TDS) of 5% to 95% comprising 0.5 % to 3.5 % of Withanolide glycoside; and
2. Prepared Hydroalcoholic extract of Ashwagandha powder comprising 0.5% to 15% of Withanolide glycoside;
3. The hydroalcoholic extract was mixed with the water extract in the ratio of 5:95 to 95:5 with continuous stirring for 10-45 minutes.
Step 3:
1. The protective coat of step 1 was mixed with soluble matrix of Ashwagandha prepared in step 2 for 10 minutes to 45 minutes at 37?
The mixture was dried using tray dryer or vacuum drier or Fluid bed dryer or spray dried using a spray drier.

Example-3: Estimation of Withanolide content in the formulation
A gravimetric method was employed to determine glyco withanolides and aglycon withanolides in various formulations. Approximately 3 grams of dry extract powder was accurately weighed and placed in a 250 mL beaker. To this, 50 mL of a methanol-water solution (50:50) was added and mixed using a magnetic stirrer. The mixture was then transferred to a separating funnel and defatted with hexane.
The hydroalcoholic layer was re-extracted using diethyl ether, and the diethyl ether layer was collected and washed with distilled water. The washed ether layer was evaporated to dryness to obtain the free withanolides, representing the aglycon component.
The remaining hydroalcoholic layer was then washed with a chloroform-methanol mixture (2:1), and evaporated to dryness, yielding the glyco withanolide fraction

Table 1.
Formulation code Aglycon withanoloide (%) Glyco withanoloide (%)
E18 1.30 1.30
E26 3.00 3.03
E30 5.90 2.03
E31 1.60 3.13
E33 2.06 0.90
E34 2.30 0.75

Example-4 Physicochemical characteristics
Different physicochemical parameters like loss on drying, pH , particle size, bulk density alcohol soluble extractive value and water soluble extractive value were carried out for the formulation.
Table 2:
E18 E26 E30 E31 E33 E34
Description Dark green granular powder Dark green granular powder Dark green granular powder Dark brown granular powder Greenish brown granular powder Dark green granular powder
Odor Characteristic Characteristic Characteristic Characteristic Characteristic Characteristic
Taste bitter bitter bitter bitter bitter bitter
Moisture Content (LOD) 6% 5.38% 4.98% 5.87% 6% 4.54%
pH (1% solution) 6.3 6.14 6.1 5.58 5.44 6.21
Particle size % (passed through 30#) 100% 97% 100% 100% 100% 100%
Bulk Density 0.41 g/ml 0.44 g/ml 0.40 g/ml 0.47 g/ml 0.44 g/ml 0.79 g/ml
Hygroscopicity not hygroscopic not hygroscopic not hygroscopic not hygroscopic not hygroscopic not hygroscopic
Alcohol soluble extractive 47.84% 45.65% 42.31% 23.53% 41.7% 40.98%
Water soluble extractive 46.98% 40.35% 42.65% 47.84% 41.05% 42.98%

Example: 5
Gastric acid-resistant profile of the developed formulation compared to Ashwagandha extract, Study-1
To evaluate the gastric acid resistance profile of the formulations in relation to the extract, a solubility study was conducted. Specifically, 1 g of the extract and formulations E18 and E33 were introduced into a freshly prepared gastric buffer (pH 1.2) and incubated in a water bath at 37°C for 1 hour, with occasional stirring. Following this incubation period, the mixture was filtered, and the release of the components was assessed visually and documented (Figure 1).
Gastric acid-resistant profile of the developed formulation compared to Ashwagandha extract, Study-2
The gastric acid resistance profile of the formulations, in comparison to the extract, was evaluated using in vitro dissolution testing in simulated gastric fluid (pH 1.2) (Electrolab Dissolution Tester, USP, TDT-06P). The dissolution medium, consisting of simulated gastric fluid without enzymes and degassed to remove dissolved air, was added to the vessel of the dissolution apparatus. The apparatus was then assembled, and the dissolution medium was heated to maintain a temperature of 36.5°C to 37.5°C. At 30-minute intervals, over a two-hour period, aliquots of the dissolution medium were withdrawn, and the release was measured by recording the absorbance at 227 nm using a UV spectrophotometer. As depicted in Figure 2, the formulations demonstrated a significantly lower absorbance value compared to the extract.
Example-6
In vitro release study
1. Calibration Curve preparation of pure Ashwagandha Extract:
For quantification of Withanolides from Ashwagandha extract by UV spectrophotometer ICH guidelines are followed. In the quantification method methanol: water (50:50) mixer used as solvent.
Method Development:
i. 50 mg of Extract was dissolved in 50 mL of prepared solvent methanol: water (50:50) to prepare a stock solution 1mg/mL (1000 µg/mL).
ii. Then a secondary stock solution was prepared from the primary stock i.e., 1mL of primary stock solution was diluted in 10mL of solvent having a concentration of (100 µg/mL).
iii. Then a scan was carried out in UV spectrophotometer to in the range of 200-800nm to obtain the wavelength maxima. (Figure-3)

Preparation of Test Solution:
i. From the secondary stock solution further dilution were carried out to prepare the calibration curve.
ii. Concentrations were prepared at a range of 5, 10, 15, 20, 25, and 30 µg/mL.
iii. The absorbance of different dilutions was carried out at 227 nm wavelength to determine the regression equation.
iv. The calibration curve was prepared in excel after putting the absorbance. (Figure-4)
Table-3: Concentration and absorbance obtained after scanning of sample in 227nm.
Concentration (µg/ml) Absorbance (nm)
0 0
5 0.298
10 0.536
15 0.772
20 0.999
25 1.262

2. Release Study:
The release pattern of formulation (E18, E33) was compared with the Ashwagandha extract by testing in simulated gastric fluid (pH 1.2) and simulated intestinal fluid (pH 6.8) by in vitro USP dissolution apparatus (Electrolab, dissolution tester,USP,TDT-06P).The dissolution medium (simulated gastric fluid without enzyme, pH 1.2), free from dissolved air, was filled into the vessel of the dissolution apparatus. Apparatus was assembled and dissolution medium was heated to 36.5° to 37.5°. After 120 minutes of operation, the dissolution medium was changed to simulated intestinal fluid with pH 6.8 and the analysis was carried out for 5 hours. Samples were collected in every 30 mins for 5 hours and estimated for the % Ashwagandha extract release by UV using the above-described method. The results are illustrated in Figure 5. The formulations demonstrated a lower release profile in gastric conditions at pH 1.2 compared to extract indicating the gastric resistant property. However, a significantly enhanced sustained release profile was observed when the medium's pH was adjusted to 6.8.

Example - 7
HPTLC fingerprinting profile:
A high-performance thin-layer chromatography (HPTLC) method was employed for the phytochemical profiling of different extracts of Ashwagandha plant part and the developed formulation. For the analysis, 1 g of the sample was soaked in 10 ml of methanol. The solution was then sonicated for 15 minutes and then centrifuged, The supernatant was used as test solution. The test sample solutions were applied at 0.2 µl in below mentioned order (Table-2).
Merck aluminium plate precoated with Silica gel 60F254 was used, and the samples were applied using a CAMAG Linomat V applicator. The plate was developed using a solvent system composed of toluene, ethyl acetate, and glacial acetic acid in a ratio of 55:45:3 (v/v/v), followed by drying.
The plate was observed using a CAMAG TLC Visualizer 3 under ultraviolet (UV) light at wavelengths of 254 nm and 366 nm, and photographs were documented at these wavelengths (figure 4). Thereafter the plate was derivatized by dipping it in sulfuric acid reagent and heated at 105°C in a hot air oven until spots appeared. The images were then documented under white light and ultraviolet (UV) light at wavelengths of 366 nm (figure 4). The Rf values and fingerprint data were recorded using the WINCATS software version 3.2 SP2.

Table-4 HPTLC sample detail
Track number Sample detail
1 Hydroalcoholic extract of ashwagandha whole plant
2 Hydroalcoholic extract of ashwagandha root
3 Hydroalcoholic extract of ashwagandha root
4 E 31
5 E 18
6 E 32
7 E 33

A significant variation was observed between Ashwagandha whole plant and root extracts on tracks 1, 2, and 3 under UV light at 254 nm. Specifically, the spots with Rf values of 0.05 and 0.51 were absent in tracks 1 and 3, but present in track 2. A similar pattern was observed in the respective formulations on tracks 5, 2, and 6. Additionally, the spots with Rf values of 0.8 and 0.9, specific to the root extract, were missing in tracks 2, 3, 4, and 6, but present in tracks 1 and 5, which correspond to the whole plant extract. Track 7, representing formulation E33, which contains both the whole plant and root extracts, exhibited bands corresponding to both the whole plant (track 1) and root (track 2) extracts. The study further indicates that the formulations displayed a similar HPTLC fingerprinting pattern when compared to the respective extracts.

Advantages of the invention:
1. Provides acid resistance coating on the Ashwagandha and thus helps in reduction of degradation of actives in stomach pH.
2. Soluble matrix of Ashwagandha in a defined concentration range and ratio of the water extract and the hydroalcoholic extract of Ashwagandha wherein the water extract has more polysaccharide which enables in entrapment of oil soluble Withanolide.
3. The protective coating comprises of lipid, polysaccharides and protein which is obtained from a natural source and hence does not make use of any synthetic polymeric material for the protective coating.
4. Composition comprising Ashwagandha extract of the present invention does not degrade at acidic pH and releases in intestinal pH.
5. The composition of the present invention provides sustained release of the active in gastrointestinal system.
6. The composition of the present invention provides Enhanced Bioavailability of Ashwagandha.
7. The composition comprising Ashwagandha extract of the present invention can be administered at lower dose like 250mg to 300mg per day.

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:
1. A composition comprising the plant extract of Ashwagandha, curcuminoid, quercetin, green tea extracts, sylemarin extract or Boswellia extract encapsulated in a natural protective coat selected from lipids, polysaccharides and protein.

2. The composition as claimed in claim 1 comprising the Ashwagandha extract containing 0.5-10%w/w of Withanolide glycoside encapsulated in a natural protective coat selected from lipids, polysaccharides and protein.

3. The composition as claimed in claim 2, wherein the Ashwagandha extract comprises the water extract of Ashwagandha root or other plant parts containing 0.5 % to 3.5 % w/w of Withanolide glycoside.

4. The composition as claimed in claim 2, wherein the Ashwagandha extract comprises the hydroalcoholic extract containing 0.5 % to 15 % w/w of Withanolide glycoside.

5. The composition as claimed in claim 2, wherein the Hydroalcoholic extract of Ashwagandha and the water extract of Ashwagandha are in the ratio of 5:95 to 95:5.

6. The composition as claimed in claim 2, wherein the lipids are selected from Medium- chain triglyceride (MCT) oil, Sunflower oil, Sea buckthorn oil, Almond oil, Olive oil, Moringa Oil, Withania Oil, Fenugreek Oil, Flaxseed oil, Cumin oil, Grape seed oil, evening prime-rose oil, Ashwagandha oil or mixtures thereof in a concentration ranging from 1 to 15% w/w.

7. The composition as claimed in claim 2, wherein the lipids contain other oils specified with Stearic acid in a concentration ranging from 0.1% to 60% w/w, Oleic acid in a concentration ranging from of 0.1 % to 90% w/w, Palmitic acid in a concentration ranging from 0.1 % to 60% w/w, Caprylic acid in a concentration ranging from 0.1 % to 60% w/w and Linoleic acid in a concentration ranging from 0.1% 60% w/w.

8. The composition as claimed in claim 2, wherein the polysaccharides are selected from natural starch in a concentration ranging from 1% to 50% w/w.

9. The composition as claimed in claim 8, wherein the natural starch is selected from Rice starch, Potato starch, Tapioca starch, Corn starch; Modified starch selected from Pregelatinized starch, Maltodextrin or mixtures thereof.

10. The composition as claimed in claim 2, wherein the Plant protein is in a concentration ranging from 1% to 75% w/w.

11. The composition as claimed in claim 10, wherein the Plant protein is selected from Rice protein, Pumpkin seed protein, Almond protein, Pea protein, oat protein, hemp seed protein, sunflower protein, mung protein, fenugreek protein or mixtures thereof.

12. The composition as claimed in claim 2, wherein the composition may comprise natural hydrocolloids, cellulose gum or emulsifier.

13. The composition as claimed in claim 12, wherein the natural hydrocolloids comprises plant pectin, Gum acacia; Guar gum, xanthan gum or mixtures thereof in a concentration ranging from 1% to 50% w/w.

14. The composition as claimed in claim 12, wherein the cellulose gum is selected from methyl cellulose; Hydroxypropyl Methyl Cellulose, ethyl cellulose or mixtures thereof.

15. The composition as claimed in claim 12, wherein the emulsifiers are selected from sunflower lecithin, soya lecithin, Datem and Plant Saponins or mixtures thereof in a concentration range of 0.5 % to 5% w/w.

16. The composition as claimed in any one of the preceding claims 1-15, wherein said composition exhibits enhanced bio-activity and bioavailabilty without degradation at gastrointestinal pH.

17. A process for preparation of the composition of encapsulated Ashwagandha extract in a natural protective coat comprising;
i. Preparing the protective coat by adding lipids in water and mixing followed by addition of emulsifiers, polysaccharides and protein and further mixing;
ii. Preparing the soluble matrix of Ashwagandha by mixing the hydroalcoholic extract of Ashwagandha powder with the water extract of Ashwagandha root or other plant part with continuous stirring; and
iii. Mixing the protective coat of step (i) with the soluble matrix of Ashwagandha of step (ii) followed by drying to obtain the desired product.
OR
iii. Homogenising Ashwagandha hydroalcoholic extract with water extract of Ashwagandha in liquid form for a specified time interval and further adding the components of the protective coat which comprise of oil, Lecithin, Polysaccharide and protein to form a slurry; and
iv. Drying the slurry.

18. The process as claimed in claim 17, wherein the active Withanolide glycoside in the extract is in the range of 0.5 to 10% w/w.

19. Use of the composition comprising the encapsulated Ashwagandha extract containing 0.5-10% w/w of Withanolide glycoside as active in a protective coat as claimed in any one of the preceding claims for sustained release of the active.

20. A method of enhancing the bioactivity and bioavailability of the Ashwagandha extract comprising administering said composition claimed in any one of the preceding claims.

21. A method of sustained release of the Ashwagandha extract in gastrointestinal medium without degradation comprising administering said composition of the Ashwagandha extract, whereby the active Withanolide glycoside in the extract of Ashwagandha is released from the protective coat at the pH of the gastrointestinal medium.