DESC:

FORM 2

THE PATENTS ACT, 1970
(39 of 1970)
AND
THE PATENT RULES, 2003

COMPLETE SPECIFICATION
(See section 10; rule 13)

“The integrated active composition for osteoarthritis and joint wellness”

OMNIACTIVE HEALTH TECHNOLOGIES LIMITED

An Indian Company, registered under the Indian Companies Act, 1956 having its registered office located at OmniActive Health Technologies Limited, T-8b, 5th Floor, Phoenix House,
A wing, Phoenix Mills Compound, 462, Senapati Bapat Marg, Lower Parel, Mumbai – 400013 Maharashtra, India.

The following specification particularly describes the invention and the manner in which it is to be performed.

FIELD OF THE PRESENT INVENTION

The present invention is related to the integrated active composition for prevention, improvement and maintenance of arthritis. More particularly, the invention is related to a composition comprising the integrated active extract and at least one pharmaceutically and/or nutraceutically accepted excipient. More specifically the invention is related to the integrated actives selected from one or more of withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia or combination thereof for improvement, prevention and maintenance of arthritis and for use in arthritis more specifically, for osteoarthritis.

BACKGROUND OF THE PRESENT INVENTION
Arthritis is a more specific term that implies damage or inflammation in one or more joints. The condition is often manifested by pain, swelling, heat, redness and limitation of movement. The most common type of arthritis is osteoarthritis or 'wear and tear arthritis'. The condition results from the wearing down of cartilage. Since the cartilage cannot be properly replaced by the body, it may make new bone at the edge of the joint to compensate for the loss of cartilage. This in turn produces bony swellings which are painful because the new bone is stretching the sensitive lining of the pre-existing bone.
Knee osteoarthritis is a type of disease that results from the disintegration of articular cartilage and underlying bones. More than 10% of the world population is affected by this disease. For example, about 14 million Americans are exposed to knee osteoarthritis. It is estimated that the knee osteoarthritis ratio of the U.S. population comprising of African Americans and Hispanics would be projected to rise from 38% in 2014 to 56% of the total population by 2060. It caused pain, stiffness, and deformation in osteoarthritis patients that severely affected quality of life. Patients and their physicians often turn to surgical treatments to address progressive pain and disability, including arthroscopy and total knee arthroplasty. However, in 6 months after total knee arthroplasty, 16–39% of patients still sustain knee pain.
In general, numerous cytokines are expressed and are functionally active in synovial tissues, and multiple factors, including NF-?B, IL-1ß and TNF-a, are associated with the pathogenesis of osteoarthritis. Increasing evidence suggests that activation of the NF-?B pathway induces the expression of inflammatory cytokines that induce the destruction of the joint, leading to onset and progression of osteoarthritis. Matrix metalloproteinases (MMPs) such as MMP3 is a major enzyme that targets cartilage for degradation. It is not only known to play crucial roles in degrading collage type II, which comprises over 90% of cartilage matrix proteins, but also degrades proteoglycan, collage type IV and type IX.
Recently, research has been focused on natural products from plants that have multifunctional anti-inflammatory effects that may decrease the toxicity and opposing effects of the drug the treatment of osteoarthritis. For instance, curcumin, a polyphenol extracted from rhizomes of turmeric and Curcuma longa, has many biological activities, such as anti-inflammatory, antioxidant, anti-cell proliferation, and antiapoptotic; and has wide sources, low toxicity, and few adverse reactions. It has been widely used in the treatment of many diseases, such as diabetes, cardiovascular diseases, autoimmune diseases, tumors, and so forth. Curcumin, the major curcuminoid (50 to 60%) is associated to two other major components, i.e., demethoxycurcumin (DMC, 2) (20 to 30%), and bisdemethoxycurcumin (BDMC, 3) (7 to 20%). DMC and BDMC play important roles in contributing to efficacy and stability of the curcumin. Natural turmeric extracts should contain curcumin, DMC and BDMC. Nevertheless, the use of curcumin has been limited due to its poor aqueous solubility, chemical instability, photodegradation, rapid metabolism, and short half-life. Therefore, it is useful to develop a new formulation containing enriched levels of BDMC.
Recently, the gum resin extracted from the ancient herb, Boswellia serrata has gained considerable attention as a potent anti-inflammatory, anti-arthritic and analgesic agent. 3-O-acetyl-11-keto-beta-boswellic acid (AKBA) is the most active compound of boswellia extract and is a potent inhibitor of 5-lipoxygenase (5-LOX), a key enzyme in the biosynthesis of leukotrienes from arachidonic acid in the cellular inflammatory cascade.
US20170112823A1 relates to compositions and methods for treating a condition associated with inflammation. In some embodiments the condition may include neuropathy or arthritis. In various embodiments, the invention teaches kits for treating a condition associated with inflammation. In some embodiments, the invention teaches kits for treating neuropathy or arthritis.
US7070814B2 relates to a bio enhancing/bioavailability-facilitating composition comprising: an effective amount of an extract and/or at least one bioactive fraction from Cuminum cyminum; one or more additive selected from drugs, nutrients, vitamins, nutraceuticals, herbal drugs/products, micro nutrients, antioxidants along with pharmaceutically acceptable additives/excipient, and optionally, an effective amount of piperine or extract/fraction of piper nigrum or piper longum; and a process for the preparation of such extracts and active fractions from plant Cuminum cyminum.

EP3417846A1 relates to a food and/or nutraceutical composition, which comprises an endocannabinoid, preferably but not limited to, Palmitoyl ethanol amide (PEA).

WO2008120220A1 relates to novel dietary supplement compositions comprising enriched 3- O-acetyl-11-keto-ß-boswellic acid and enriched demethylated curcuminoids, wherein the composition exhibits anti-inflammatory, antiulcerogenic and antioxidant activities. Also disclosed are novel compositions comprising enriched 3-0-acetyl-ll-keto-ß- boswellic acid, enriched demethylated curcuminoids that have a synergistic effect on specific inhibition of- COX-2 and 5-LOX activity and optionally containing glucosamine, resveratrol, garlic extract, chondroitin, methyl sulphonyl methane, bromelain, serrati peptidase, quercetin, gallic acid, caffeic acid, green tea extract, aspirin and ibuprofen.

US5494668A relates to a method of treating degenerative musculoskeletal diseases such as rheumatoid arthritis and osteoarthritis in an animal, typically a human, comprises administering to the animal, typically enterally, in a convenient dosage form, a therapeutically effective amount of the beneficiated extracts of the plants ASHWAGANDHA (Withania somnifera), SALLAI GUGGUL (Boswellia serrata), TURMERIC (Curcuma longa), and GINGER (Zingiber officinal) in a predetermined proportion relative to each other with or without other biologically active inorganic ingredients, such as zinc sulphate.

By inspecting all the prior arts are completely silent over the composition of selective withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia or combination thereof for improvement, prevention and maintenance of arthritis more specifically osteoarthritis. So, there is long felt need to provide selective nutritional composition for prevention, improvement and maintenance of osteoarthritis.

The present invention relates to composition in the form of the single integrated active composition containing withanolide glycoside enriched ashwagandha, BDMC enriched curcumin and AKBA enriched boswellia used for arthritis more specifically for osteoarthritis.

OBJECTIVES OF THE PRESENT INVENTION

The primary objective of the present invention is to develop the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and at least one pharmaceutically and/or nutraceutically accepted excipients.

The primary objective of the present invention is to develop the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and at least one pharmaceutically and/or nutraceutically accepted excipients use for the prevention, improvement and maintenance of arthritis and more specifically for osteoarthritis and joint wellness.

Another objective of the present invention is to develop the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and at least one pharmaceutically and/or nutraceutically accepted excipients use for the prevention, improvement and maintenance of osteoarthritis.

The further objective of the present invention is to provide the integrated active composition which exhibits increased joint mobility, comfortable movements for ease of daily activities, improve walking performance, increased joint flexibility, reduction in joint stiffness, reduced joint discomfort/pain, reduced inflammation and reduced cartilage breakdown.

Another objective of the present invention is to develop the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and at least one pharmaceutically and/or nutraceutically accepted excipients use for the prevention, improvement and maintenance of rheumatoid arthritis.

The another objective of the present invention is to provide the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia in selective percentage which is formulated in different forms like powder, beadlets, granule, capsules, tablets, oil suspension, films or any other suitable oral or topical dosage forms using pharmaceutically and/or nutraceutically acceptable excipients.

Further objective of the present invention is to provide a process of preparation of the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and at least one pharmaceutically and/or nutraceutically accepted excipients.

SUMMARY OF THE PRESENT INVENTION:

According to an aspect of the present invention there is provided an integrated active composition comprising:
a) Withanolide glycoside enriched ashwagandha in the range of 10-50% w/w of the composition;
b) Bisdemethoxycurcumin(BDMC) enriched curcumin in the range of 20-60% w/w of the composition;
c) Acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia in the range of 5-30% w/w of the composition and
d) At least one or more pharmaceutically or neutraceutically accepted ingredient.

Further the present invention is to develop the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and at least one pharmaceutically and/or nutraceutically accepted excipients use for the prevention, improvement and maintenance of arthritis and more specifically for osteoarthritis and joint wellness.

BRIEF DESCRIPTION OF ACCOMPANYING FIGURES

Figure 1. Effects of integrated active composition (IAT) on IL-ß (A), IL-6 (B), TNF-a (C) and NF-?ß (D) levels in the knee joint of monosodium iodoacetate (MIA) induced osteoarthritis (OA) rats. The bars point out the standard deviation of the mean. Asterisks above the lines indicate statistical differences among the groups (ANOVA and Turkey's post-hoc test; *P<0.05; ***P<0.001; **** P<0.0001). IL-1ß, interleukin-1ß; IL-6, interleukin-6; TNF- a, tumor necrosis factor a; NF-?ß, nuclear factor kappa ß.

Figure 2. Effects of integrated active composition (IAT) on Collagen Type 2 (A), MMP-3 (B), COX-2 (C) and LOX-5 (D levels in the knee joint of monosodium iodoacetate (MIA) induced osteoarthritis (OA) rats. Asterisks above the lines indicate statistical differences among the groups (ANOVA and Turkey's post-hoc test; *P<0.05; **** P<0.0001); MMP-3, matrix metalloproteinase-3; COX-2, cyclooxygenase-2; LOX-5, 5-lipoxygenase.

Figure 3. Effects of integrated active composition (IAT) on the knee joint of monosodium iodoacetate (MIA) induced osteoarthritis (OA) rats. Representative radio graphic images (A) obtained at the end of the experiment are shown. Mean values of Kellgren- Lawrence scores are demonstrated (B) with ± standard deviations. Asterisks above the line indicate statistical differences among the groups represented (B) with bar (Kruskal-Wallis followed by Mann-Whitney U; *P<0.05; **P<0.01; compared as OA group).

Figure 4. Effects of integrated active composition (IAT) on histopathology of the knee joint of monosodium iodoacetate (MIA) induced osteoarthritis (OA) rats. Representative histopathologic images of hematoxylin-eosin (A) and toluidin blue (B) staining obtained at the end of the experiment are shown. Mean values of Mankin scores are demonstrated with ± standard deviations. Asterisks above the line indicate statistical differences among the groups represented (C) with bar (Kruskal-Wallis followed by Mann-Whitney U; *P<0.05; compared as OA group).

Figure 5. Effects of integrated active composition (IAT) on knee swelling (A), left knee joint diameter (B) right knee joint diameter (C) and the ratio of right to left diameter values in monosodium iodoacetate (MIA) induced osteoarthritis (OA) rats. Asterisks above the lines indicate statistical differences among the groups (ANOVA and Turkey's post-hoc test; *P<0.05; **P<0.01; ***P<0.001; **** P<0.0001).

Figure 6. Effects of integrated active composition (IAT) on paw area (B) and stride length (D) in monosodium iodoacetate (MIA) induced osteoarthritis (OA) rats. Representative measures of paw area (A) and stride length (C) are shown. Asterisks above the lines indicate statistical differences among the groups (ANOVA and Turkey's post-hoc test; *P<0.05; **P<0.01; ***P<0.001; **** P<0.0001).

DETAILED DESCRIPTION OF THE PRESENT INVENTION

The invention described herein relates to the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and at least one pharmaceutically and/or nutraceutically accepted excipients use for prevention, improvement and maintenance of arthritis more specifically to osteoarthritis and joint wellness.

Within the context of this invention the terminology the “integrated active composition (IAT)” is commonly used in the specification to refer composition having selectively added active ingredients withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin, acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and one or more of pharmaceutically and/or nutraceutically accepted excipients which promote enhancement in result.

Withania somnifera is cultivated in many of the drier regions of India. Withania somnifera, known commonly as ashwagandha, Indian ginseng, poison gooseberry or winter cherryis a plant in the Solanaceae or nightshade family. The main phytochemical constituents are withanolides – which are triterpene lactones.

Within context of the composition “withanolide glycoside enriched ashwagandha extract” is further defined as the extract of ashwagandha with selectively to make rich or richer especially by the addition or increase of desirable withanolide glycosides like 1a,3ß,22R)-3-[(6-O-ß-D-Glucopyranosyl-ß-D-glucopyranosyl)oxy]-1,22,27-trihydroxy-ergosta-5,24-dien-26-oic acid d-lactone (Withanolide IV),
1a,3ß,22R)-3-[(6-O-ß-D-Glucopyranosyl-ß-D-glucopyranosyl)oxy]-1,22-dihydroxy-ergosta-5,24-dien-26-oic acid d-lactone (Withanolide V), 2R)-2-[(1R)-1-hydroxy-1-[(1S,3R,8S,9S,10R,13S,14S,17S)-1-hydroxy-10,13-dimethyl-3-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-[[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]oxan-2-yl]oxy-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl]ethyl]-4,5-dimethyl-2,3-dihydropyran-6-one (Withanolide VI- ) as per the composition.
Curcumin (1, 7 bis (4-hydroxy-3-methoxy phenyl)-l, 6 heptadiene-3, 5- dione), is a principal curcuminoid of the popular Indian spice Curcuma longa commonly known as turmeric, a perennial herb of Zingiberaceae (ginger) family. Curcumin is derived from the rhizome of Curcuma longa and has been traditionally used in the treatment of skin wounds, inflammation, tumors etc. Within context of the composition “bisdemethoxycurcumin (BDMC) enriched curcumin” is further defined as curcumin with selectively to make rich or richer especially by the addition or increase of desirable curcuminoids such as bisdemethoxycurcumin as per the composition.
Boswellia serrata is a plant that produces Indian frankincense. It is also known as Indian oli-banum, Salai guggul, and Sallaki in Sanskrit. The plant is native to much of India and the Punjab region that extends into Pakistan. Boswellia serrata contains chemical constituents, such as ß-boswellic acid, acetyl-ß-boswellic acid, 11-keto-ß-boswellic acid and acetyl-11-keto-ß-boswellic acid. Within context of the composition “acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia ” is further defined as boswellia with selectively to make rich or richer especially by the addition or increase of desirable acetyl-11-keto-ß-boswellic acid (AKBA) as per the composition.
According to embodiment of the present invention the integrated actives composition comprising ashwagandha, curcumin, boswellia and one or more of pharmaceutically and/or nutraceutically accepted excipients.

According to further embodiment of the present invention the integrated active composition comprising ashwagandha is enriched with withanolide glycoside.

According to further embodiment of the present invention the integrated active composition comprising ashwagandha enriched with withanolide glycoside in the range of 1-40%.

According to further embodiment ashwagandha enriched with withanolide glycoside is present in the range of 20-40% w/w of the composition.

According to further embodiment of the present invention the integrated active composition comprising curcumin enriched with bisdemethoxycurcumin (BDMC).

According to further embodiment of the present invention the integrated active composition comprising curcumin extract contains bisdemethoxycurcumin (BDMC) in the range of 5-50%.

According to further embodiment the bisdemethoxycurcumin (BDMC) enriched curcumin extract is present in the range of 30-60% w/w of the composition.

According to further embodiment of the present invention the integrated active composition comprising boswellia extract enriched with acetyl-11-keto-ß-boswellic acid (AKBA).

According to further embodiment of the present invention the integrated active composition comprising boswellia extract contains acetyl-11-keto-ß-boswellic acid (AKBA) in the range of 70-90 %.

According to further embodiment the acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia extract is present in the range of 5-20 % w/w of the composition.

As per one more embodiment, at least one more excipient employed in the composition may be selected from, but not limited to hydrophilic carrier, an antioxidant, pH modifier, solvents, solublizer, binder, bioavailability enhancer, fat and the like or the combinations thereof.

In one more embodiment, the hydrophilic carrier employed in preparation of the integrated active composition is selected from the group such as, but not limited to, cellulose derivatives, polyacrylates, polyethylene glycols, povidones, starch and starch derivatives, gums, sugars, pectin, plant polysaccharides, protein such as casein, whey protein, soy protein, pea protein and the like. The compositions may/may not be free of povidone as hydrophilic carrier.

In one more embodiment, the cellulosic polymer in preparation of the integrated active composition is selected from the group such as, but not limited to Hydroxypropyl methyl cellulose, microcrystalline cellulose and/or combination thereof.

According to preferred embodiment the hydrophilic carrier is Hydroxypropyl methyl cellulose.
According to further embodiment, the hydrophilic carrier is present in the range of 5-20 % w/w of the composition.

In one more embodiment, the bioavailability enhancer in preparation of the integrated active composition is selected from the group such as, but not limited to d-limonene, monoglycerides, phospholipids such as lecithin/phosphatidyl choline, botanical extracts, and/or combination thereof. The bioavailability enhancers also act as a solubility enhancer.

According to main embodiment of the present invention the bioavailability enhancing agent is lecithin and the source for the lecithin preferably from sunflower.

According to further embodiment the bioavailability enhancer is present in the range of 1- 10% w/w of the composition.

In one more embodiment, the solvent in preparation of the integrated active composition is selected from the group such as, but not limited to, isopropyl alcohol, acetone, methanol, alcohol, ethyl acetate, ethanol, methylene dichloride, water.

In one more embodiment, the solubilizer in preparation of the integrated active composition is selected from the group such as, but not limited to propylene glycol alginate, sugar alcohols, sugar esters, phospholipid, Vitamin E TPGS (d-a-Tocopheryl polyethylene glycol 1000 succinate), beta cyclodextrin and/or combination thereof.

In one more embodiment, the binder in preparation of the integrated active composition is selected from the group such as, but not limited to hydroxy propyl cellulose, pregelatinized starch and/or combination thereof.

In one more embodiment, the pH modifier in preparation of the integrated active composition is selected from the group such as, but not limited to Citric acid, trisodium citrate, lactic acid, L-arginine, Calcium carbonate, magnesium carbonate, and/or combination thereof.

In one more embodiment, the antioxidant in preparation of the integrated active composition is selected from the group such as, but not limited to natural tocopherols, ascorbyl palmitate, rosemary extract, epigallocatechin gallate, catechins, ascorbic acid and/or combination thereof.

According to further embodiment, the integrated active composition described herein exhibit enhanced bioavailability and the compositions can be available in orally administrable solid, semisolid, liquid forms, selected from, but not limited to dosages such as, powders, granules, pellets, beadlets, caplets, tablets, capsules, soft gel capsules, solution, emulsions, suspensions, oil suspensions, dispersions and the like.

According to one more embodiment of the present invention the integrated active composition is in the form of granule.

According to one more embodiment of the present invention the integrated active composition is in the form of powder.

According to embodiment of the present invention, a process for the preparation of the integrated active composition comprises:
a) Dissolve one or more excipients in water /other solvent(s) or mixture thereof which is
previously heated with continuous stirring till it completely dissolves;
b) Add curcumin extract, ashwagandha extract and Boswellia extract with continuous
stirring followed by homogenization;
c) Shift the dispersion obtained in step (b) through mesh to obtain particles of desired size;
and
d) Further spray drying of dispersion obtained from step (c).

The primary objective of the present invention is to develop the integrated active composition comprising withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia use for the prevention, improvement and maintenance of arthritis more specifically osteoarthritis and joint wellness.

According to further embodiment of the present invention the integrated active composition comprising of withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia for the prevention, improvement and maintenance of osteoarthritis.

According to further embodiment of the present invention the integrated active composition comprising of withanolide glycoside enriched ashwagandha, bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia for the prevention, improvement and maintenance of joint wellness.

According to further embodiment of the present invention the integrated active composition comprising of ashwagandha , bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia for the prevention, improvement and maintenance of rheumatoid arthritis.

According to further embodiment of the present invention the integrated active composition which exhibits increased joint mobility, comfortable movements for ease of daily activities, improve walking performance, increased joint flexibility, reduction in joint stiffness, reduced joint discomfort/pain, reduced inflammation and reduced cartilage breakdown.

According to further embodiment of the present invention the integrated active composition comprising of ashwagandha extract, bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia for the improvement in muscle strength and muscle performance, reduction in muscle soreness and muscle damage and promote faster muscle recovery.

Example 01:
Ingredient Quantity of Ingredients (mg) Quantity of Ingredients (%)
withanolide glycoside enriched Ashwagandha 138.89 30.19
BDMC enriched Curcumin 210.53 45.77
AKBA enriched Boswellia 58.83 12.79
HPMC (hydroxypropyl methyl cellulose) 42.55 9.25
Lecithin 9.20 2.00
Water 3780.00 821.74
460.00 100.00

The process for preparation of composition has defined in the stepwise manner as follows :
Step 1: Weigh batch quantity of water and add to vessel
Step 2: Heat water to 80 °C
Step 3: Add batch quantity of Lecithin into step 2 under stirring at 500-800 RPM until lecithin is completely dissolved
Step 4: Add batch quantity of HPMC into step 3 under stirring at 800-1200 RPM until HPMC is completely dissolved
Step 5: Add batch quantity of BDMC enriched curcumin extract, withanolide glycoside enriched ashwagandha extract, AKBA enriched boswellia extract to step 4 under stirring at 1000-1500 RPM until a uniform dispersion is obtained
Step 6: Sift the step 5 dispersion through # 20 mesh (ASTM) before spray drying
Step 7: Spray drying of dispersion obtained from step 5.

At the outset of the description that follows, it is to be understood that the ensuing description only illustrates a particular form of this invention. However, such a particular form is only an exemplary embodiment and is not intended to be taken restrictively to imply any limitation on the scope of the present invention.

By careful inspection of the prior arts available in public domain, all the arts are completely silent over on the claimed composition wherein the some of the arts are with singly ingredient merely speaks about osteoarthritis without any experimentation support like laundry basket selection. But none of the prior art teaches or suggest about the composition of this instant invention.

Here we are discussing one by one in comparison with this instant invention.In M. Z. Siddiqui et al. Indian J Pharm Sci. 2011 May-Jun; 73(3): 255–261), this published article describes resinous part of Boswellia serrata possesses monoterpenes, diterpenes, triterpenes, tetracyclic triterpenic acids and four major pentacyclic triterpenic acids i.e. ß-boswellic acid, acetyl-ß-boswellic acid, 11-keto-ß-boswellic acid and acetyl-11-keto-ß-boswellic acid, responsible for inhibition of pro-inflammatory enzymes. Out of these four boswellic acids, acetyl-11-keto-ß-boswellic acid is the most potent inhibitor of 5-lipoxygenase, an enzyme responsible for inflammation .

But the literature does not provide any supporting data showing effect of selectively enriched acetyl-11-keto-ß-boswellic acid obtained from boswellia for use of osteoarthritis.

In International Immunopharmacology (Volume 7, Issue 4, April 2007, Pages 473-482) explains pure compound from Boswellia serrata extract exhibits anti-inflammatory property in human PBMCs and mouse macrophages through inhibition of TNFa, IL-1ß, NO and MAP kinases.

This art is completely silent on the experimental data showing the effect of the acetyl-11-keto-ß-boswellic acid on various symptoms of the osteoarthritis.

In K .N. Pushpakumari International journal of pharmaceutical sciences and research a projected impact factor (2019): 1.230 cite score (2017): 0.27), The comparative data of bio-absorption of two turmeric formulations containing curcuminoids in a varied composition compared to regular turmeric extract.

The document is completely silent over the effect of bisdemethoxy curcumin on osteoarthritis.
A pilot prospective study (Indian J Med Res. 2015 Jan; 141(1): 100–106) explains Efficacy & safety evaluation of Ayurvedic treatment (Ashwagandha powder & Sidh Makardhwaj) in rheumatoid arthritis patients but does not explain the effect of specific ashwagandha extract on osteoarthritis.

Clinical experimentation study
Objective: The present study has been assessed for the efficacy of integrated active composition (test product) in a monosodium iodoacetate (MIA) induced osteoarthritis.

Test product: The composition containing BDMC enriched curcumin, AKBA enriched boswellia and withanolide glycoside enriched Ashwagandha is manufactured by OmniActive Health Technologies Limited, India.

Dose 1: 100 mg/kg of formulation
Dose 2: 200 mg/kg of formulation.
Experimental Design: Female wistar rats (8 weeks) randomly allocated into groups as defined below (n=7 in each):
I. Normal Control,
II. Osteoarthritis group,
III. Osteoarthritis, BDMC enriched curcumin, AKBA enriched boswellia and withanolide glycoside enriched ashwagandha dose1 (100 mg/kg of formulation) group, and
IV. Osteoarthritis, BDMC enriched curcumin, AKBA enriched boswellia and withanolide glycoside enriched ashwagandha dose2 (200 mg/kg of formulation).

In order to induce osteoarthritis rat model, the right knee of the rats shaved and disinfected with 70% alcohol following anaesthetization using with xylazine (10 mg/kg) and ketamine hydrochloride (50 mg/kg). 3 mg of MIA dissolved in 50 µL saline and injected into right knee joints through the infrapatellar ligament using a 0.3 ml insulin syringe fitted with a 29-G needle. Control group received an injection of 50 µL saline. Two weeks after injection with MIA, the products dissolved in 1 mL saline orally given for 4 weeks. All rats observed every other alternate day to assess knee joint swelling.

Four weeks after given product, the rats sacrificed, and blood and the specimens of the knee joint collected for the follow-up experiment. The blood samples centrifuged at 3,000 rpm for 10 min, and the harvested sera kept at -20 °C until the day of analysis.

Integrated active sample preparation:
Integrated active compositions according to the present invention are as follows:
The test sample for 100 mg/kg & 200 mg/kg has prepared as per the example 1 with the process parameters provided.

Biochemical Analysis
Serum of MDA (malondialdehyde) analyzed by HPLC (high performance liquid chromatography). Antioxidant enzymes (SOD, CAT, GSHPx), IL-1ß, IL-6, IL-10, and TNF-a measured using the relevant commercial kits according to the enzyme-linked immunosorbent assay (ELISA) method.

Western Blot Analysis
Articular cartilage samples analyzed for the expression of TNF-a, IL-6, IL-1ß, nuclear factor kappa B, cyclooxygenase-2 (COX-2), collagen type2, IL-1B, CRP, 5-LOX, MMP3, Cartilage oligomeric matrix protein (COMP) using the Western blot technique.

Histological Analyses

Histological changes assessed to confirm the effects of product on cartilage degeneration in the knee joints of MIA-induced osteoarthritis rats. Following the rat sacrifice, each knee joint respected, fixed in 10% formalin (Sigma-Aldrich; Merck KGaA) for 24 h at 4°C, and decalcified with 5% hydrochloric acid (Sigma-Aldrich; Merck KGaA) for 4 days at 4°C. Following decalcification specimens dehydrated in graded acetone and embedded in paraffin. Sections (thickness, 2–3 µm)stained with 0.2% hematoxylin and 1% eosin (H&E; Sigma-Aldrich; Merck KGaA) for 5 min and 3 min, respectively. The histological preparations were analyzed and photographed with a microscope using a digital image capture camera.

Example 1
The objective of the study has demonstrated the synergistic effect of integrated active composition as per this invention on serum inflammation markers.

Method: Animals were sacrificed, and serum samples collected and assessed for serum biomarkers measured using the relevant commercial kits according to the enzyme-linked immunosorbent assay (ELISA) method as shown in the table 1 and illustratively provided in fig 1.

Table 1:
Items Groups --P--*
Control OA OA+JHF1 OA+JHF2
TNF-a, pg/mL 17.19±3.06d 43.17±4.21a 33.96±3.87b 26.61±3.46c 0.0001
IL-1ß, pg/mL 9.65±1.61d 40.83±3.99a 29.44±3.85b 22.22±4.22c 0.0001
IL-6, pg/mL 65.26±6.97a 28.08±5.99c 36.13±4.60c 47.03±6.42b 0.0001
IL-10, pg/mL 29.61±5.73d 82.51±8.79a 55.12±7.21b 43.12±6.83c 0.0001
COMP, ng/mL 9.26±0.98d 31.97±3.60a 21.93±1.83b 14.94±1.89c 0.0001
CRP, mg/L 1.94±0.11c 10.13±1.74a 4.72±0.38b 3.61±0.75b 0.0001

Abbreviations:
OA: Osteoarthritis group
OA+ JHF1: Osteoarthritis, BDMC enriched curcumin, AKBA enriched boswellia
and withanolide glycoside enriched ashwagandha dose1 (100 mg/kg
of formulation) group
OA+ JHF2 : Osteoarthritis, BDMC enriched curcumin, AKBA enriched boswellia
and withanolide glycoside enriched ashwagandha dose1 (200 mg/kg
of formulation) group

TNF-a: tumor necrosis factor a; IL-1ß: interleukin-1ß; IL-6: interleukin-6; IL-10: interleukin-10; COMP: cartilage oligometrix matrix protein; CRP: c-reactive protein. Statistical comparisons are indicated with different superscript (a-d) in the same row (P < 0.05; *ANOVA and Turkey's post-hoc test). Mean values of items are demonstrated with ± standard deviations.

Conclusion:
From the table 1 and fig 1, it was observed that serum inflammatory markers such as TNFa, IL1ß, IL10, COMP which are responsible for induction of inflammation and associated pathology were increased during OA and treatment with JHF 1 and JHF 2 reduced these levels in a statistically significant manner. Levels of cytokine IL-6 were reduced during OA were moderately restored by treatment. CRP levels increased during inflammatory conditions were also reduced to normal levels during treatment.

Example 2
The objective of the study was to demonstrate synergistic effect of integrated active composition on serum antioxidant markers.

Method: Animals were sacrificed, and serum samples collected and assessed for serum antioxidant markers. Serum of MDA was analyzed by HPLC. Antioxidant enzymes (SOD, CAT, GSHPx) were measured using the relevant commercial kits according to the enzyme-linked immunosorbent assay (ELISA) method and data shown in the table below.

Table 2:
Items Groups --P--*
Control OA OA+JHF 1 OA+ JHF 2
MDA, nmol/mL 0.58±0.09c 2.08±0.35a 1.31±0.17b 0.89±0.11c 0.0001
SOD, U/mL 60.27±9.82a 23.51±3.10c 34.33±2.76b 42.27±4.71b 0.0001
CAT, U/mL 152.40±5.58a 92.77±5.39d 114.88±7.53c 130.04±4.69b 0.0001
GSH-Px, U/mL 96.38±5.53a 38.36±3.03d 58.34±5.12c 74.65±5.70b 0.0001
MDA: malondialdehyde; SOD: superoxide dismutase; GSH-Px: glutathione peroxidase; CAT: catalase.

Statistical comparisons are indicated with different superscript (a-d) in the same row (P < 0.05; *ANOVA and Turkey's post-hoc test). Mean values of markers are demonstrated with ± standard deviations

Conclusion:
MDA which is a marker of oxidative stress that play a key role in induction of pathology of OA was significantly increased during OA and were significantly reduced during treatment. Similarly, treatment significantly improved the levels of antioxidant enzymes, SOD, CAT and GSH-Px as shown in the table 2.

Example 3
The objective of the study was to demonstrate synergistic effect of integrated active composition on inflammation markers in synovial joint tissue.

Method: Animals were sacrificed, and synovial tissue samples collected and assessed for inflammatory markers by western blot as shown in figure 01.

Conclusion:
It was observed that protein levels of inflammatory markers as shown below; IL-ß (A), IL-6 (B), TNF-a (C) and NF-?ß (D) were increased in OA rats as measured by western blot followed by densitometric analysis of the relative intensity according to the control group after ß-actin normalization to ensure equal protein loading. Blots were repeated at least three times (n?=?3) and a representative blot (E) is shown. It was observed that treatment with IAT at two doses (100 & 200 mg/kg) reduced the levels of inflammatory markers in the joint tissue in a statistically significant manner.

Example 4
The objective of the study was to demonstrate synergistic effect of integrated active composition on inflammation markers in synovial joint tissue.

Method: Animals were sacrificed, and synovial tissue samples collected and assessed for markers of cartilage degradation and inflammation by western blot as shown in figure 02.

Conclusion:
It was observed that protein levels of markers of collagen degradation such as Collagen Type 2 (A), MMP-3 (B) and inflammatory markers COX-2 (C) and LOX-5 (D) shown below were increased in OA rats as measured by western blot followed by densitometric analysis of the relative intensity according to the control group after ß-actin normalization to ensure equal protein loading. Blots were repeated at least three times (n?=?3) and a representative blot (E) is shown. It was observed that treatment with IAT at two doses reduced the levels of markers of cartilage degradation and inflammation in the joint tissue in a statistically significant manner

Example 5
The objective of the study was to demonstrate synergistic effect of joint health formula ingredients as a combination on joint architecture of synovial joint as assessed by X-ray image analysis

Method: Animals were evaluated for structural integrity of synovial joint by radiography analysis as shown in figure 03 and table 04.

Table 4. Kellgren- Lawrence scoring system (Kellgren and Lawrence, 1957).

Stage Radiologic Findings
0 None
1 Doubtful: Suspicious narrowing of the joint space and possible osteophyte formation.
2 Minimal: Definite osteophyte and possible narrowing of the joint space.
3 Moderate: Numerous moderate osteophytes, definite narrowing of the joint space, some sclerosis and possible deformity of the bone ends.
4 Severe: Large osteophytes, marked narrowing of the joint space, sclerosis and deformity of the bone ends

Conclusion:
It was observed that induction of OA is associated with loss of joint structural integrity as observed by radiographic images (A) obtained at the end of the experiment. Significant improvement in the joint architecture was observed after treatment with JHF 1 & JHF 2. The above results were further validated by measuring mean values of Kellgren- Lawrence scores (B) with ± standard deviations.

Example 6
The objective of the study was to demonstrate synergistic effect of joint health formula ingredients as a combination on morphological analysis of synovial joint tissue.

Method: Animals were sacrificed, and synovial tissue samples collected, fixed in 10% formalin for 24 h at 4°C, decalcified with 5% hydrochloric acid for 4 days at 4°C, dehydrated in graded acetone and embedded in paraffin. Sections of 2–3 µm thickness were stained with 0.2% hematoxylin and 1% eosin for 5 min and 3 min, respectively and histological preparations were analyzed and photographed with a microscope using a digital image capture camera as shown in figure 04.

Conclusion:
It was observed that there was a significant loss of joint structure and inflammatory infiltration of joint in OA condition as observed after histopathologic images of hematoxylin-eosin (A) and toluidine blue (B) staining obtained at the end of the experiment. This was further validated by mean values of Mankin scores (C).

Example 7
The objective of the study was to demonstrate synergistic effect of integrated active composition on morphological analysis of synovial joint tissue.

Method: Knee was visually assessed for morphological changes of joint pathology such as knee swelling, knee joint diameter, and ratio of right to left diameter values as shown in figure 05.

Conclusion:
Both right and left knee joint swelling was observed when OA was induced in rats and this was significantly reduced by treatment with JHF 1 & JHF 2 (A), This was further validated by measuring left knee joint diameter (B) right knee joint diameter (C) and the ratio of right to left diameter values and a significant treatment effect was observed with JHF 1 & JHF 2.

Example 8
The objective of the study was to demonstrate synergistic effect of joint health formula ingredients as a combination on morphological analysis of synovial joint tissue.

Method: Knee was visually assessed for morphological changes of joint pathology such as paw area and stride length as shown in figure 06.

Conclusion:
Effects of integrated active composition on paw area (B) and stride length (D) in monosodium iodoacetate (MIA) induced osteoarthritis (OA) rats. Representative measures of paw area (A) and stride length (C) are shown.

Based on the results and conclusions provided on osteoarthritis model, we here by presume that the same integrated active composition will have a similar effect on muscle health. The cell line study model with further evidenced by the study protocol has been illustrated below to study the efficacy on muscle health.

Further study to demonstrate the effect of combination of the integrated active selected from one or more of withanolide glycoside Ashwagandha , bisdemethoxycurcumin (BDMC) enriched curcumin and acetyl-11-keto-ß-boswellic acid (AKBA) boswellia on muscle health will be done as following.

In-Vitro Study:

Objective: Demonstration of ergogenic potential integrated active composition (test product) for improving physiological or metabolic responses that may help in muscle development (prevent loss of muscle protein as in age-related sarcopenia), enhance exercise performance, ameliorate muscle injury and muscle inflammation (DOMS)
Cell model: C2C12 murine myoblasts

Experimental approaches: We are in process with the basic assays and generate more information after we identify selected ingredients for development C2C12 myoblasts was cultured at 37°C in humidified 5% CO2 were plated in 24 well-plate and incubated for 24h before adding treatment.
1. Cytotoxicity assay
2. Mitochondrial density assay: Effect of ingredients on mitochondrial biogenesis is analyzed by measuring cellular respirations accompanied by increase in mitochondrial mass is calculated.
3. Antioxidant Capacity: After 24h of treatment, the total antioxidant capacity was determined in C2C12 homogenates
4. Lactate dehydrogenase and Creatine Kinase activity: LDH and CK activities are used as indicator of the occurrence of muscle cell damage and is assessed by determining the release of the cytosolic enzyme lactate dehydrogenase (LDH) and creatine kinase (CK) activity
5. IGF-1 Elisa assay: IGF-I exerts acute anabolic actions on protein and carbohydrate metabolism by increasing the cellular uptake of amino acids in cells and plays a major role in the regulation of skeletal muscle growth.

In-Vivo Study:

Objective: To evaluate the effect of integrated active composition (test product) formulation on endurance, grip strength and muscle metabolism in rats.

Animals and Experimental Design:
21 Wistar Albino or Sprague Dawley male rats (20% extra animals), age: 8 week, weight: 180 ± 20 g will be housed in a controlled environment with a 12:12-h light-dark cycle at 22°C and will be provided with rat chow and water ad libitum.
Rats will be randomly divided into 3 treatment groups, each containing 7 animals.
1. Control (Normal and exercised)
4. JHF 1 (100 mg/kg of formulation)
5. JHF 2 (200 mg/kg of formulation)
At screening period, the grip strength will be performed on day -3. On day -2, Treadmill Test will be performed.

Grip Strength Test:
The grip strength of the rats will be evaluated using a force measurement system. Combined forelimb and hindlimb grip strength and forelimb grip strength will be measured at the end of the oral administration period. The system has an electronic digital force gauge that determines the peak force. Each rat will be held by the tail until it released the pull bar. Five consecutive tests will be performed on each rat to obtain the peak value.
Treadmill Test:
An animal treadmill will be used to measure the running endurance of the rats. The rats will be ran on the treadmill at a speed of 12 m/min on 0° incline to become acclimated to the test in advance. In the actual test, the rats will be ran at the speed of 12 m/min on 0° incline followed by an increase of 3m/min every 20 min thereafter. After 60 min, the incline will be increased by 5° every 20min. The shock grid will be set to deliver 0.2 mA of electricity, which did not physically injure the animals. The time at which the rats will be unable to run after 10 s of electric shock will be defined as exhaustion.

At the end of the study, all rats will be subjected to overnight fasting and sacrificed by cervical dislocation and blood and gastrocnemius muscle were collected. Serum samples will be obtained by taking blood samples to gel biochemical tubes after centrifugation (5000 rpm at 4°C for 10 minutes). Samples of the liver and from the gastrocnemius muscle (taken from approximately the same location each time) will be quickly removed, placed on ice, and kept at -80°C until analyses. For biochemical assays, tissues samples will be homogenized within 10 min in 10 volumes of cold Tris 10 mM (pH 7.4). Muscle homogenates will be centrifuged at 4000 ×g at 4 °C for 10 min to yield the low-speed supernatant fraction that was used for the lipid peroxidation analyses.

Biochemical Analysis:
Serum glucose, lipid profile, aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine levels will be analyzed with a portable automated chemistry analyzer (Samsung LABGEO PT10V, Samsung Electronics Co., Suwon, Korea). Rat Lactate Assay Kit (Cayman Chemical Co., Ann Arbor, MI, USA) will be used to measure the serum lactate concentrations through enzyme-linked immunosorbent assays (ELISA, Elx-800, Bio-Tek Instruments Inc, Vermont, USA). ELISA (MyBioSource, San Diego, CA, USA) will also be used in measuring serum myoglobin concentration.

The malondialdehyde (MDA) level in muscle tissue will be measured by high-performance liquid chromatography (Shimadzu, Tokyo, Japan) using a Shimadzu UV-vis SPD-10 AVP detector and C18 ODS-3, 5 µm, 4.6 mm ×250 mm column. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) will be determined using commercially available kits (Cayman Chemical, Ann Arbor, MI, USA) according to the manufacturer’s procedure.

It has to evident to those skilled in the art that the invention is not limited to the details of the illustrative examples and that the present invention may be embodied in other specific forms without departing from the essential attributes thereof. It is therefore desired that the present embodiments and examples be considered in all respects as illustrative and not restrictive, reference being made to the appended claims, rather than to the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

,CLAIMS:We claim:

1. An integrated active composition comprising;
a) Withanolide glycoside enriched ashwagandha;
b) Bisdemethoxycurcumin (BDMC) enriched curcumin;
c) Acetyl-11-keto-ß-boswellic acid (AKBA) enriched boswellia and
d) at least one or more pharmaceutically or nutraceutically accepted ingredient.

2. The integrated active composition as claimed in claim 1, wherein the pharmaceutically or nutraceutically accepted ingredient is selected from hydrophilic carrier, bioavailability enhancer, an antioxidant, pH modifier, solvents, solublizer, binder, fat or the combinations thereof.

3. The integrated active composition as claimed in claim 1, wherein the hydrophilic carrier is selected from Hydroxypropyl methyl cellulose, microcrystalline cellulose and/or combination thereof, polyacrylates, polyethylene glycols, povidones, starch and starch derivatives, gums, sugars, pectin, casein, whey protein, soy protein, pea protein in the range of 5 to 20% by weight of total composition.

4. The integrated active composition as claimed in claim 1, wherein the bioavailability enhancer and/or solubility enhancer is selected from d-limonene, monoglycerides, phospholipids such as lecithin/phosphatidyl choline, or botanical extracts in the range of 1 to 10% by weight of total composition.

5. The integrated active composition as claimed in claim 1, wherein the composition is in the form of powders, granules, pellets, beadlets, caplets, tablets, capsules, soft gel capsules, solution, emulsions, suspensions, oil suspensions, dispersions.

6. The integrated active composition as claimed in claim 1, wherein the composition is in the form of powders, granules and/or the combinations thereof.

7. The process for preparing the integrated active composition as claimed in claims 1 to 7 comprising steps of:

(i) dissolving one or more excipients in water /other solvent(s) or mixture thereof
which is previously heated with continuous stirring till it completely dissolves;
(ii) adding curcumin extract, ashwagandha extract and Boswellia extract with
continuous stirring followed by homogenization;
(iii) sifting the dispersion obtained in step (ii) through mesh to obtain particles of
desired size; and
(iv) further spray drying of dispersion obtained from step (iii) to obtain integrated
active composition.

8. The integrated active composition as claimed in claim 1, wherein the composition is use for prevention, improvement and maintenance of pre-osteoarthritis and/or osteoarthritis and joint wellness.

9. The integrated active composition as claimed in claim 8, wherein the composition is use for prevention, improvement and maintenance of rheumatoid arthritis.

10. The integrated active composition as claimed in claim 8, wherein the composition increases joint mobility, comfortable movements for ease of daily activities, improve walking performance, increased joint flexibility, reduction in joint stiffness, reduced inflammation and reduced cartilage breakdown.

11. The integrated active composition as claimed in claim 8, wherein composition decrease tumor necrosis factor (TNF), interleukin1ß (IL-1ß), interleukin-10 (IL-10), Collagen Type 2, matrix metalloproteinase-3 (MMP3), cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX), cartilage oligometrix matrix protein (COMP); c-reactive protein(CRP), malondialdehyde (MDA).

12. The integrated active composition as claimed in claim 8, wherein composition increases interleukin-6 (IL-6),superoxide dismutase (SOD), glutathione peroxidase(GSH-Px), catalase (CAT).

13. The integrated active composition as claimed in claim 1, wherein the composition is use for prevention, improvement and maintenance of muscle strength, muscle performance, muscle soreness, muscle recovery and muscle damage.

14. The integrated active composition as claimed in claim 1, wherein the subject is human or animal.

15. The integrated active composition as claimed in claim 1, wherein dose is 10-500 mg/kg.

Dated this 30th November 2020.

Applicant
OmniActive health Technologies Ltd
Dr Pravin Shejul