# US 11,304,927 B2 · Bioactive extract, fraction of Cassia occidentalis and formulation thereof for bone regeneration

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- Number: US 11,304,927 B2
- Document ID: US11304927B2
- Status: Granted
- Kind: B2
- Application number: 16/434,323
- Filed: 2019-06-07
- Published: 2019-12-12
- Granted: 2022-04-19
- Earliest claimed: 2018-06-08
- Assignees: Council of Scientific and Industrial Research
- Applicants: Council of Scientific and Industrial Research
- Inventors: Naibedya Chattopadhayay; Subhashis Pal; Sudhir Kumar; Ramakrishna Eppalapally; Padam Kumar; Sapana; Jiaur Rahaman Gayen; Riyazuddin Mohammed; Sabyasachi Sanyal; Anagha Gurjar; Prabhat Ranjan Mishra; Naresh Mittapelly; Kamal Ram Arya; Brijesh Kumar; Srikanta Rath; Arun Kumar Trivedi; Maurya Rakesh
- Web page: https://patentgrep.com/patent/US11304927B2

## CPC

- A61K 31/353: 3,4-Dihydrobenzopyrans, e.g. chroman, catechin
- A61K 9/0053: {Mouth and digestive tract, i.e. intraoral and peroral administration}
- A61K 9/08: Solutions {(composition of solutions A61K47/00)}
- A61K 9/1075: {Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers (A61K9/0026 takes precedence)}
- A61K 31/7048: having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin {, digitoxin or digoxin}
- A61K 36/482: Cassia, e.g. golden shower tree
- A61K 47/10: Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
- A61K 47/44: Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin (synthetic glycerides, e.g. medium-chain triglycerides, A61K47/14)
- A61K 47/46: Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
- A61P 19/08: for bone diseases, e.g. rachitism, Paget's disease
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present invention relates to a bioactive extract, fraction of Cassia occidentalis and formulation thereof for bone regeneration and treatment of glucocorticoid induced musculo-skeletal diseases and associated electrolyte imbalances.

## Claims

### Claim 1

A formulation for fracture healing and protection against corticosteroid-induced musculo-skeletal diseases, wherein the formulation comprises Cassia occidentalis extract and/or bioactive fraction optionally along with one or more pharmaceutically acceptable excipients, wherein the formulation comprises alcoholic extract and/or bioactive fraction in an amount ranging from 2-10% w/w, oil in an amount ranging from 20-60% w/w, surfactant in an amount ranging from 20-40% w/w, co-surfactant in an amount ranging from 12-28% w/w, solubilizer in an amount ranging from 5-10% w/w, and excipients in an amount ranging from 10-60% w/w.

### Claim 2 (depends on 1)

The formulation as claimed in claim 1 wherein the oil is oleic acid, peanut oil, linoleic acid, soya bean oil, or a combination thereof.

### Claim 3 (depends on 1)

The formulation as claimed in claim 1 wherein the surfactant and cosurfactant is polysorbate 80, polysorbate 20, polysorbate 60, medium-chain triglycerides of caprylic (C8) and capric (10) acids, polyoxyl 35 castor oil, diethylene glycol monoethyl ether, propylene glycol monocaprylate 90, tocopherol polyethylene glycol succinate (TPGS), poly(ethylene glycol)-block-poly(propylene glycol) (poloxamer 407), propylene carbonate, polyethylene glycol, or 1,2, ethanediol.

### Claim 4 (depends on 1)

The formulation as claimed in claim 1 wherein the solubilizer is polyethylene glycol, with different molecular weights ranging from 200-5000, glycerol, caprylcaproyl macogol glycerides, lauroglycol, soya lecithin, egg lecithin, cholic acid and deoxycholic acid, polyvinyl caprolactam-polyvinyl acetate polyethylene glycol graft copolymer, sorbitan monolaurate 20, or sorbitan monolaurate 80.

### Claim 5 (depends on 1)

The formulation as claimed in claim 1 wherein the one or more excipients are polymers of sugars or inorganic materials.

### Claim 6 (depends on 1)

The formulation as claimed in claim 1 wherein the formulation is useful for treatment of fracture healing and corticosteroid-induced musculo-skeletal diseases and associated electrolyte imbalances.

### Claim 7 (depends on 1)

A process for preparation of the formulation as claimed in claim 1 wherein the process comprises the steps of:
a. percolating powdered Cassia occidentalis plant material with alcohol for a period ranging between 20 to 24 hrs followed by collecting the percolate,
b. repeating the step (a) for 4 to 5 times to obtain the alcoholic extract,
c. fractioning the alcoholic extract as obtained in step (b) with n-hexane to obtain hexane soluble fraction and hexane insoluble residue,
d. triturating the hexane insoluble residue as obtained in step (c) with ethylacetate to obtain ethylacetate soluble fraction and ethylacetate insoluble residue,
e. suspending the ethylacetate insoluble residue as obtained in step (d) with water followed by extracting with n-butanol to obtain n-butanol soluble fraction,
f. isolating Apigenin, 4-methoxy-2′,4′-dihydroxy chalcone), 7,4′-dihydroxy flavone, Luteolin, 7,3′,4′-trihydroxy-flavone, Emodin, Nicotinic acid, Chrysophanol 1-O-β-Gentiobioside, Rhamnocathartin, Isovitexin or Apigenin 6c-glucoside from the n-butanol soluble fraction as obtained in step (e) by chromatographic methods,
g. solubilizing Cassia extract/fraction obtained in step (b) or in step (e) with surfactant mixture under stirring to obtain a nanoemulsion liquid preconcentrate formulation,
h. converting the emulsion liquid preconcentrate of step (g) into a solid system in situ to obtain the formulation by mixing with a solid carrier wherein the weight ratio of liquid in-situ nanoemulsion system to solid carrier is from 1:0.5 to 1:10.

### Claim 8 (depends on 7)

The process as claimed in claim 7 wherein the alcohol is selected from ethanol or butanol.

### Claim 9 (depends on 7)

The process as claimed in claim 7 wherein the surfactant mix consists of surfactant 20-40% w/w and co-surfactant 5-10% w/w.

### Claim 10 (depends on 7)

The process as claimed in claim 7 wherein the solid carrier is selected from the group consisting of HPMC, soluplus, colloidal silica, hypermellose, and aerosil.

### Claim 11 (depends on 1)

The formulation as claimed in claim 1 wherein the formulation has an effective osteoinductive and a skeletal preservation dose is reduced to 50 mg/kg in a butanolic fraction formulation from 250 mg/kg in an ethanolic extract.

### Claim 12 (depends on 1)

The formulation as claimed in claim 1 wherein the formulation has an effective muscle protective dose is reduced to 50 mg/kg in a butanolic fraction formulation from 250 mg/kg in an ethanolic extract.

### Claim 13 (depends on 1)

The formulation as claimed in claim 1 wherein the formulation is a bioactive fraction and the relative bioavailability (as shown by AUC) of apigenin 6-C-glucoside/isovitexin (biomarker) in the fraction is enhanced by more than 5 fold as compared to the extract.

### Claim 14 (depends on 5)

The formulation of claim 5, wherein the one or more excipients are a polymer of a sugar selected from the group consisting of hypermellose, aeosil, HPMC, and soluplus and/or an inorganic material selected from the group consisting of colloidal silica, calcium carbonate, and calcium phosphate.

### Claim 15 (depends on 1)

The formulation of claim 1, wherein the extract/bioactive fraction contains marker compounds Apigenin and Isovitexin.

### Claim 16 (depends on 15)

The formulation of claim 15, wherein Apigenin is present in an amount of 0.0005% to 0.002% and Isovitexin is present in an amount of 0.0006% to 0.0008%.