DESC:FIELD OF INVENTION
The present invention is related to herbal composition of withanolides and process for preparation thereof. In particular the present disclosure solubility improvisation of high concentration of withanolides i.e. 25% from Withania somnifera (Ashwagandha). The invention further relates to phyto complex by unique OEM solubilization technique. Another invention discloses that standard withanolides are converted into complex form & further effectiveness of solubility and absorption is developed by adding a synergy form of natural celery saponin. The invention also relates to the process of solubilization and granulation to improve solubility, micromeritics property, and particle size uniformity for good therapeutical action and dosage strength.

BACKGROUND OF INVENTION
Ashwagandha is one of the rasayana herbs in Ayurveda—one of the herbs that purportedly promotes youth and longevity and alleviates suffering. It is thought to be especially rejuvenating for men; to strengthen bone marrow, muscle, and semen; and to imbue the user with the intellectual facility, in addition to long life and youthful vitality. However, it also is believed to be quite helpful to the elderly by providing energy and relieving pain, inflammation, and nervous debility.

Withania somnifera (Ashwagandha) which is also known as Indian ginseng is an evergreen herb under the traditional science of India. Withania somnifera is historically proven for a sleep disorder, rheumatism, immunity, vitality, and vigor. Its root is especially beneficial for loss of memory, mass gain, endurance, strength, and overall health. The key constituent of Withanolides glycoside has a potential role in different chronic diseases either mono or synergy form. Every botanical highly concentrated active chemical (Phytochemical) contains poor characteristics. Ashwagandha extract comprises enriched withanolide glycosides and Saponin with a negligible amount of alkaloid, and oligosaccharides. Accordingly, it is an object of this invention to provide a new and improved Withania Somnifera extract composition, having an enhanced level of withanolide glycosides and oligosaccharides, and minimum amounts of polysaccharides and free withaferin A (aglycone), and an improved extraction process for obtaining such compositions. The leaves are used to remove worms and also help to reduce inflammation, and swelling. In countries where the Ayurvedic system of medicine is officially recognized and practiced such as in the USA, Europe, China, and Gulf countries, powdered dried root of Ashwagandha is available in its supplement form.

US20170066797A1 provides a novel class of withanolides that have been isolated from W. somnifera under aeroponic conditions or produced semi-synthetically from withanolide natural products. The invention also provides pharmaceutical compositions thereof and methods for using the same in proliferative diseases, neurodegenerative diseases, autoimmune, and inflammatory diseases.

CA2991530C discloses healthful supplements are described herein. For example, the supplements can include at least four of the following types of nutrients: an amino acid blend, a coconut extract, a vegetable glycerin extract, an alcohol blend, a cannabinoid blend, a hemp plant extract, a Boswellia serrata extract, a curcuma / turmeric blend, black cumin seed, an Artemisia ludoviciana extract, an Astragalus extract, a fenugreek extract, a mushroom extract blend, or any combination thereof.

US20210369804A1 provides to an enriched Withania somnifera (Ashwagandha) extract composition comprising withanolide glycosides, withanolide aglycones and reduced levels of withaferin-A. The invention also provides to process for the preparation of these compositions and further provides to methods of improving testosterone levels, energy levels, sustained energy, vigor, stamina, and muscle mass and muscle strength using these compositions.

Progress in the Chemistry of Organic Natural Products (Vol. 94 pp 127–229) Withanolides and Related Steroids (Misico, Rosana I., et al.) discloses isolation of the first withanolides in the mid-1960s, over 600 new members of this group of compounds have been described, with most from genera of the plant family Solanaceae. The basic structure of withaferin A, a C28 ergostane with a modified side chain forming a d-lactone between carbons 22 and 26, was considered for many years the basic template for the withanolides. Nowadays, a considerable number of related structures are also considered part of the withanolide class; among them are those containing ?-lactones in the side chain that have come to be at least as common as the d-lactones. The reduced versions (? and d-lactols) are also known.

Analytical chemistry and microbiology (Volume 101Issue 61 November 2018) Extraction Optimization for Phenolic- and Withanolide-Rich Fractions from Withania somnifera Roots (Kumar, Satyanshu, et al.) Both the roots and leaves of Withania somnifera are products of commerce. They contain active compounds of therapeutic value and mostly different withanolides. Several pharmacological activities of W. somnifera have links to one or more withanolides. The presence of phenolic compounds in extracts could play a vital role in the reduction of blood glucose levels in diabetic subjects.

OBJECTIVE OF INVENTION
Phytochemistry is a wide branch and nowadays trend of nutraceuticals with plant-based botanical ingredients and the science behind converting modified dosage forms is developing. Instead of Raw herbs, recommendations of extracts from the manufacturer and even customers are increased due to quick action and its new trend. Some drawback is also there in botanical extract either phytoconstituents or phytochemicals. As the concentration of active constituent increases, solubility and bioavailability will be decreased. Different pharmaceutical excipients and solubilizers study is there to enhance the poor characteristics. But it will impact the size and strength of the overall active ingredient in the finished dose.

The present invention developed some new strategy that solves the poor characteristic without excipients/ chemicals and it also solves the other issues related to active phytoconstituent. In the present invention relates to represent withanolides isolated from Ashwagandha, a famous plant in India for various activities. High concentration of withanolides glycosides is poorly soluble in an aqueous medium due to its poor characteristic. Enteric-coated technology is there to target dosage form in an alkaline medium. But our main target is to avoid the use of chemicals and solvents used in enteric-coating technology. The activity of withanolide glycosides even at lower doses is amazing at alkaline PH or by the administration of an enteric coated formulation of extract of Ashwagandha to humans.

This protects the composition from hydrolysis in the acidic environment of the stomach to release the withanolide glycoside in neutral/alkaline pH in the gastrointestinal tract (GIT) thus absorption will be enhanced and the therapeutical effect will be high. Further, the process of preparation of the extract of Ashwagandha enriched with withanolide glycosides and saponins are disclosed, which is a modified withanolides causality-based target complex instead of a chemical enteric coated formulation.

SUMMARY OF THE INVENTION
The present invention is related to herbal composition of withanolides and process for preparation thereof.

In particular the present disclosure solubility improvisation of high concentration of withanolides i.e. 25% from Withania somnifera (Ashwagandha).

The invention further relates to phyto complex by unique OEM solubilization technique. Another invention discloses that standard withanolides are converted into complex form & further effectiveness of solubility and absorption is developed by adding a synergy form of natural celery saponin.

The invention also relates to the process of solubilization and granulation to improve solubility, micromeritics property, and particle size uniformity for good therapeutical action and dosage strength.

The causality of Phytochemical is dependent on the solubility and bioavailability of the same and it will directly connect with the therapeutically active compound. In the pharmaceutical industry, different solubilizers are employed to enhance the solubility but after solubilization, the active concentration of API dilutes, and due to this high concentration is required for the finished dose. If solubility and bioavailability are poor then it will affect the protein binding and therapeutical effect. On the other hand, excipients and chemical additives which are going to be used as solubilizers are not safe for long-term use. To overcome this drawback, Phyto complex by unique OEM solubilization technique is very important to abolish the use of excipients and avoid dose dilution. Most of the biologically active constituents of plants are polar or water-soluble molecules, but as increased with active constituent’s concentration, solubility and bioavailability can decrease.

However, water-soluble phytoconstituents (like flavonoids, tannins, glycosidic aglycones, etc. are poorly absorbed either due to their large molecular size which cannot be absorbed by passive diffusion or due to their poor lipid solubility; severely limiting their ability to pass across the lipid-rich biological membranes, resulting poor bioavailability. Standard Withanolides are converted into complex form & further effectiveness of solubility and absorption is developed by adding a synergy form of natural celery saponin. There is no interaction found between withanolides and celery mixed complex and it is confirmed by FTIR, and flowability is increased is confirmed by XRD, and micromeritics property.

DETAILED DESCRIPTION OF THE INVENTION
Detailed Description of the Invention While this specification concludes with claims particularly pointing out and distinctly claiming that, which is regarded as the invention, it is anticipated that the invention can be more readily understood through reading the following detailed description of the invention and study of the included examples.
The following is a detailed description of embodiments of the present disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.

Unless the context requires otherwise, throughout the specification which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense that is as “including, but not limited to.”

Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

OEM solubilization technique is very important to abolish the use of excipients and avoid dose dilution. Most of the biologically active constituents of plants are polar or water-soluble molecules, but as increased with active constituent’s concentration, solubility and bioavailability can decrease.

The headings and abstract of the invention provided herein are for convenience only and do not interpret the scope or meaning of the embodiments.

Various terms are used herein. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of.
Withanolides solubility is more in the solvent than aqueous medium. In hot water, solubility is more than in normal water. PH also plays an important role in the solubility of withanolides. Most of the ashwagandha extract-based products are available in capsule, powder and tablet dosage forms. Few of available in enteric-coated form because at 7.2 or 7.4 PH solubility of withanolides are more than Gastric PH. Withanolides glycoside concentration is used between 2.5- 75% i.e., isolated from Ashwagandha (Withania somnifera). As the concentration of active constituent increases, aqueous solubility will be less. Hence to replace enteric coated solvent and chemical-based process, here the designed a causality-based target complex with natural saponin i.e., Celery extract containing a high amount of saponin that helps to improve the solubility of withanolides without interaction. Solid dispersion granulation of withanolides and Celery saponin evaluated for solubility & micromeritics properties. The Withanolides complex is developed by optimized synergy complex of celery saponin by solid dispersion technology. Solubility study in different PH is analyzed compared with standard Withanolides. Standard withanolides compared with modified withanolides for micromeritics property, particle size, and flowability parameters. It is concluded, modified withanolides have better flowability than standard withanolides. Apart from it, it studied modified withanolides in various formulations and the effectiveness of the same in different clinical studies, case studies, and invitro analyses, and it seems more potent and contains good efficacy than standard withanolides.

Effect of PH on the solubility of 25% Withanolides
Table 1: Effect of PH on the solubility of 25% Withanolides
Active constituents PH Solubility (mg/ml)
Withanolides-25% 1.2 0.2 mg/ml
Withanolides-25% 4.5 0.22 mg/ml
Withanolides-25% 6 0.24 mg/ml
Withanolides-25% 7.2 0.36 mg/ml
Withanolides-25% Distilled water 0.30 mg/ml

• With PH modification in 25% Withanolides aqueous solution, solubility is slightly altered, but does not improve significantly. The highest solubility is found at PH-7.2.
• At intestinal PH, solubility is increased up to 0.06 mg/ml, which means it will be not impactful for further formulation study.

Method preparation of Withanolides Solid dispersion form
Table 2: Preparation ratio of 25% withanolides with different additives
Active constituents Additives Ratio (%)
Withanolides-25% Sodium lauryl sulfate (SLS) 1:01
Withanolides-25% Poloxamer 407 1:0.3
Withanolides-25% Beta Cyclodextrin 1:1
Withanolides-25% Celery extract (Saponin) 1:0.1

• 25% Withanolides mixed with Sodium lauryl sulfate (SLS), Poloxamer 407, Beta cyclodextrin, and Saponin of Celery extract by Solid dispersion technique for solubility enhancement. The ratio of individual additives with 25% withanolides is selected after optimization.

Solubility Enhancement in withanolides with different solubilizers
Table 3: Solubility enhancement in withanolides with different solubilizers
Active constituents
Of Synergy blend Ratio (%) Pre-Solubility
Modified solubility
Withanolides-25%: Sodium lauryl sulfate (SLS) 1:0.1 0.3 mg/ml 0.312 mg/ml
Withanolides-25%: Poloxamer 407 1:0.3 0.3 mg/ml 0.375 mg/ml
Withanolides-25%: Beta cyclodextrin 1:1 0.3 mg/ml 0.426 mg/ml
Withanolides-25%: Celery extract 1:0.1 0.3 mg/ml 0.597 mg/ml

• Pre-solubility in the aqueous phase is 0.30 mg/ml as mentioned in the effect of PH on withanolides table.
• Aqueous solubility of 25% withanolides is optimized with Sodium lauryl sulfate (SLS) (1:0.1), Poloxamer 407 (1:0.3), Beta cyclodextrin (1:1) and Celery extract (1:0.1). After solid dispersion of above additives with 25% withanolides, it is found 0.597mg/ml from 0.3 mg/ml almost double improvement in Withanolides-25%: Celery extract solid dispersion comparatively good and significant than other additives at very low concentration.
• It is concluded that at a very low concentration of celery saponin, the solubility of 25% withanolides improves double time and is far better than other additives and techniques.

Micromeritic Property Enhancement
Table 4: Micromeritic Properties of withanolides with different additives
Micromeritics properties Withanolides-25% Withanolides-25%: Sodium lauryl sulfate (SLS), Withanolides-25%: Poloxamer 407 Withanolides-25%: Beta cyclodextrin Withanolides-25%: Celery extract
Angle of Repose 41.25 36-33 34-36 33-35 31-33
Carr’s index 19.51 17-19 16-18 15-18 14-16
Hausner Ratio 1.39 1.20-1.22 1.31-1.34 1.25-1.28 1.16-1.18

• Micromeritics property of 25% Withanolides with Sodium lauryl sulfate (SLS), Poloxamer 407, Beta cyclodextrin and Celery extract contain active saponin analyzed. It is found excellent improvement in the Angle of repose, Carr’s index, and Hausner ratio in withanolides: celery extract as compared to other solubilizer complexes.
• It is also evaluated on capsule filling and tablet compression. Filling of modified withanolides is 20% improved during capsule filling. With overall excellent micromeritics properties, good flow from the hopper during tablet compression is found and improvement is found in physical evaluation parameters of the tablet like weight variation, friability, compaction of tablet and physical appearance (aesthetic look) of the finished tablet.

Formulation development of withanolides tablet
Table 5: Different formulations of withanolides tablet
Ingredients F1 (mg) F2(mg) F3(mg) F4(mg) F5(mg)
Withanolides-25% 500 500 500 500 500
Celery Saponin extract 5 - - - -
Sodium lauryl sulfate (SLS) - 5 - 5 -
Poloxamer 407 - - 15 10 -
Beta-cyclodextrin - - - - 500
Lactose - - - - 40
Sodium Starch Glycolate - 10 - - 20
Starch Paste - - - - 20
Tack 20 10 10 10 50
Total weight of Tablet 525 525 525 525 1150

• 25% Withanolides formulation is optimized with different solubilizers, super disintegrants, modifying diluents, lubricants and binders. The active weight of 25% withanolides maintains the same in all formulations and the ratio of solubilizer keeps maintained as per the previous solubility study. Additional other excipients like diluent, binder, lubricant and super disintegrant are evaluated for solubility enhancement, moisture content, dosage release, friability, hardness, disintegration time and aesthetic appearance of the tablet.

Physical evaluation of Optimized batch
Table 6: Physical evaluation of Optimized batch
Physical evaluation F1 F2 F3 F4 F5
% Release in 45 min 98.13 68.12 93.98 56.18 79.98
Moisture content Less than 1 % Less than 1 % Less than 1 % Less than 1 % Less than 1 %
Disintegration time % 90-120 sec 290-320 sec 490-620 sec 220-320 sec 240-360 sec
Friability 0.810 0.986 1.105 0.939 1.012
Enhanced Solubility 49.6 % Less than 5% Less than 10% Less than 20% Less than 10%

• Optimized formulation F1 proven best as per Physical evaluation of finished tablet. As per solubility analysis, 25% Withanolides: Celery (Saponin extract) gives the highest solubility as compared to other solubilizers. Same concentration ratio is selected in F1 formulation without other excipients. Improvement is found throughout the evaluation parameters.
• In F1 optimized batch- the % Release in 45 min is more than 90% i.e., 98.13 comparable good to other formulations. As with disintegration time, friability, and aesthetic appearance are comparable good than other formulations. It is concluded, the solubility of 25% withanolides also affects the finished formulation.

Comparative Evaluation with Marketed formulation
Table 7: Comparative Evaluation with Marketed formulation
Physical evaluation Optimized batch Marketed -1 Marketed-2
% Release in 45 min 98.13 68.12 72.98
Disintegration time (Sec) 90-120 sec 290-320 sec 490-620 sec
Hardness 3-4 4-5 5-5.5
Friability 0.810 0.989 1.201
Finished 520 mg 500 mg 600 mg

• Finished optimized Physical evaluation parameters are compared with the marketed product of Ashwagandha extract containing withanolides. The disintegration time is 1.5-2 minutes, almost comparatively very less than the marketed formulation. Friability with adequate hardness is also relatively better than marked formulation. % Release of withanolides in 45 min is almost 30% more than the marketed formulation. It is concluded that this research is therapeutically and efficacy-wise more impactful as per market need.

,CLAIMS:1. An herbal composition of withanolides and process for preparation thereof.
2. The herbal composition of withanolides as claimed in claim 1, wherein solubility improvisation of high concentration of withanolides i.e. 25% from Withania somnifera (Ashwagandha).
3. The herbal composition of withanolides as claimed in claim 1, wherein withanolides are converted into complex form & further effectiveness of solubility and absorption is developed by adding a synergy form of natural celery saponin.
4. The herbal composition of withanolides as claimed in claim 1, wherein preparation of withanolides phyto complex by unique OEM solubilization technique.
5. The herbal composition of withanolides as claimed in claim 1, wherein process of solubilization and granulation to improve solubility, micromeritics property, and particle size uniformity for good therapeutical action and dosage strength.
6. The herbal composition of withanolides as claimed in claim 1, wherein 25% Withanolides mixed with Sodium lauryl sulfate (SLS), Poloxamer 407, Beta cyclodextrin, and Saponin of Celery extract by Solid dispersion technique for solubility enhancement
7. The herbal composition of withanolides as claimed in claim 1, wherein formulation development of withanolides tablet comprising of Withanolides-25%, Celery Saponin extract, Sodium lauryl sulfate (SLS), Poloxamer 407, Beta-cyclodextrin, Lactose, Sodium Starch Glycolate, Starch Paste, Tack.