PREPARATION THEREOF.

PRIORITY:

This application claims the benefit of Indian complete application number 202021008974 dated 2 ^nd March 2020 entitled, ‘Liposomal compositions and process for preparation thereof and 202122007627 dated 23 ^rd Feb 2021 entitled, ‘Liposomal compositions of curcumin and process for preparation thereof, the contents of which are incorporated herein by reference.

Technical field of the invention:

The present invention relates to a liposomal composition comprising an active ingredient, lipids as encapsulation material and a solvent, wherein the encapsulation material and solvent are present in a weight ratio of 1:1 to 1:5 and active ingredient and solvent are present in a weight ratio of 1 : 1 to 1 :5.

Background of the invention:

Vitamin C, also known as ascorbic acid and l-ascorbic acid, is a vitamin found in various foods and sold as a dietary supplement. It is used to prevent and treat scurvy. Vitamin C is an essential nutrient involved in the repair of tissue and the enzymatic production of certain neurotransmitters. It is required for the functioning of several enzymes and is important for immune system function. It also functions as an antioxidant.

Vitamin C is an essential nutrient for certain animals including humans. The term vitamin C encompasses several vitamins that have vitamin C activity in animals. Ascorbate salts such as sodium ascorbate and calcium ascorbate are used in some dietary supplements. These release ascorbate upon digestion. Ascorbate and ascorbic acid are both naturally present in the body, since the forms interconvert according to pH. Oxidized forms of the molecule such as dehydroascorbic acid are converted back to ascorbic acid by reducing agents. It is found in fresh fruits, berries and vegetables. It is one of the water-soluble vitamins. Without enough vitamin C, a person can get a sickness called scurvy. Lack of vitamin C was a serious health problem on long ocean trips where supplies of fresh fruit were quickly used up. Most animals make their own vitamin C.

Vitamin C, also known as L-ascorbic acid, has been known to be a very important cosmetic active for a long time. It functions as an anti-oxidant, anti inflammatory, a tyroinase inhibitor (blocking the production of melanin), and a collagen synthesis material. These multi-functional benefits is the reason that vitamin C is so dearly desired to be in cosmetic products. Unfortunately, it is also one of the most unstable materials as well. Vitamin C quickly degrades creating color and odor issues making sale of products containing it not desirable. The industry has created many derivatives to get around this issue. They have not totally been successful either. Limitations on concentration and aesthetics are still a big concern.

Janelle L. Davis et al; Nutr Metab Insights. 2016; 9: 25-30 discloses ‘Liposomal-encapsulated Ascorbic Acid: Influence on Vitamin C Bioavailability and Capacity to Protect Against Ischemia-Reperfusion Injury’. In this article, Intravenous administration of vitamin C has been shown to decrease oxidative stress and, in some instances, improve physiological function in adult humans. Oral vitamin C administration is typically less effective than intravenous, due in part to inferior vitamin C bioavailability. The purpose of this study was to determine the efficacy of oral delivery of vitamin C encapsulated in liposomes. On 4 separate randomly ordered occasions, 11 men and women were administered an oral placebo, or 4 g of vitamin C via oral, oral liposomal, or intravenous delivery. The data indicate that oral delivery of 4 g of vitamin C encapsulated in liposomes (1) produces circulating concentrations of vitamin C that are greater than unencapsulated oral but less than intravenous administration and (2) provides protection from ischemia- reperfusion-mediated oxidative stress that is similar to the protection provided by unencapsulated oral and intravenous administrations. Luka Wechtersbach; LWT - Food Science and Technology, Volume 45, Issue 1, January 2012, Pages 43-49 discloses Liposomal stabilization of ascorbic acid in model systems and in food matrices. In this study, Ascorbic acid (AA) was encapsulated into dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylcholine/cholesterol (DPPC/chol) liposomes. The study showed that co-encapsulation of citric and ascorbic acids into liposomes results in considerable stabilization of the latter in model systems containing catalytic concentrations of copper ions. The rate of AA oxidation was decreased by up to 300-fold that of free AA. Incorporation of cholesterol into membranes resulted in lower stabilization efficacy at room temperature and better thermal stability at higher temperatures than pure DPPC liposomes. Total AA, extra-liposomal AA and leakage of AA out of liposomes during thermal treatment were evaluated using the free radical 2,2'-azino-bis(3- ethylbenzthiazoline-6-sulphonic acid) (ABTS). A modification of the ABTS based assay for liposome permeability makes it applicable to reducing agents and antioxidants as well as AA. Encapsulated AA was also stabilized in real food matrices. The rate of AA oxidation in apple juice, relative to that of free AA, was decreased by two orders of magnitude when encapsulated in DPPC liposomes and by more than one order of magnitude in DPPC/chol liposomes. Liposomal stabilization of AA in a fermented milk product was less pronounced.

Coenzyme Q10 is a vitamin-like substance found throughout the body, but especially in the heart, liver, kidney, and pancreas.

Coenzyme Q10 is most commonly used for conditions that affect the heart such as heart failure and fluid build-up in the body (congestive heart failure or CHF), chest pain (angina), and high blood pressure. It is also used for preventing migraine headache, Parkinson disease, and many other conditions.

Coenzyme Q10 is an important vitamin-like substance required for the proper function of many organs and chemical reactions in the body. It helps provide energy to cells. Coenzyme Q10 also seems to have antioxidant activity. People with certain diseases, such as heart failure, high blood pressure, gum disease, Parkinson's disease, blood infections, certain diseases of the muscles, and HIV infection, might have lower levels of coenzyme Q10.

It is chemically 2-[(2E,6E,10E,14E,18E,22E,26E,30E,34E)-3,7, 11,15,

19,23,27,31 ,35,39- Decamethyltetraconta-2,6, 10, 14, 18,22,26,30,34,38- decaenyl]-5,6-dimethoxy-3-methylcyclohexa-2,5-diene-1 ,4-dione with the following structure.

Curcumin [1, 7-bis (4-hydroxy-3-methoxyphenyl)-1 , 6-heptadiene-3,5-dione] is the major yellow pigment of turmeric, a commonly used spice, derived from the rhizome of the herb Curcuma longa Linn. Curcumin have many scientifically-proven health benefits, such as the potential to prevent heart disease, Alzheimer's and cancer. It is a potent anti-inflammatory and antioxidant and also helps to improve symptoms of depression and arthritis.

In the Indian subcontinent and Southeast Asia, turmeric has traditionally been used as a treatment for inflammation, skin wounds, and tumors. In preclinical animal models, curcumin has shown cancer chemo preventive; antineoplastic and anti-inflammatory properties. Curcumin blocks the formation of reactive- oxygen species, possesses anti-inflammatory properties as a result of inhibition of cyclooxygenases (COX) and other enzymes involved in inflammation; and disrupts cell signal transduction by various mechanisms including inhibition of protein kinase C. These effects may play a role in the agent's observed antineoplastic properties, which include inhibition of tumor cell proliferation and suppression of chemically induced carcinogenesis and tumor growth in animal models of cancer.

Curcumin is an alternative to non-steroidal anti-inflammatory drugs (NSAIDS), which also have anti-inflammatory and cancer chemopreventive properties. This is so because COX is a bifunctional enzyme with cyclooxygenase and peroxidase activities. The peroxidase function contributes to the activation of procarcinogens. Therefore, the failure of NSAIDS to inhibit the peroxidase function of COX potentially limits their effectiveness as anticancer agents. Curcumin, in contrast, down-regulates levels of COX-2 and thereby decreases both the cyclooxygenase and peroxidase activities of the enzyme.

Curcumin is among the few agents to block both the COX and LOX (lipoxygenase) pathways of inflammation and carcinogenesis by directly modulating arachidonic acid metabolism. Although administered curcumin has poor bioavailability and only low or non-measurable blood levels were observed (Perkins, S. et al, Cancer Epidemiol. Biomark. Prev., 2002, 11:535- 40), this route of administration inhibits chemically induced skin and liver carcinogenesis (Limtrakul, P., et al, Cancer Lett., 1997, 116:197-203; Chiang, S. E. et al, Carcinogenesis, 2000, 21:331-35). Oral administration of curcumin also inhibits the initiation of radiation-induced mammary and pituitary tumors (Inano, H. et al, Carcinogenesis, 2000, 21:1835-41; Int. J. Radiat. Oncol. Biol. Phys., 2002, 52:212-23; ibid, 2002, 53:735-43). Similarly, in a study to assess the curcumin levels in the colorectum, a daily dose of 3.6 g curcumin achieves pharmacologically effective levels in the colorectum with negligible distribution of curcumin outside the gut.

Liposome is a tiny bubble (vesicle) made out of the same few component materials of cell membrane. Liposomes can be filled with drugs and used to deliver drugs for cancer and other diseases. Liposomes are technically simple or double emulsions in submicron size generally 20 - 250 u in diameter. The following are some of the liposomal formulations available under the trade name AmbBiosome, Abelcet, Amphotec, Doxil, DaunoXome. The methods of preparation of liposome include mechanical dispersion method, solvent dispersion method and detergent removal method.

So there is a need for more stable, liposomal composition which will increase the bioavailability and targeting property of the active ingredient and to increase the dissolution rate of the oil/water soluble drug.

The present invention is a liposomal composition and a process for preparation thereof for enhancing bioavailability, stability and improving the targeting property in a colloidal drug delivery system where is in solid dispersion form.

Liposomes are manufactured microscopic, hollow spherical vesicles composed of a lipid bilayer. When loaded with pharmaceuticals and/or dietary supplements, liposomes are a very effective method of drug/supplement delivery. When ingested, the pharmacokinetic properties of liposome intestinal absorption override the usual absorption pattern of the encapsulated drug. That is, the delivery of a drug/supplement with a typically slow or regulated pattern of absorption, such as vitamin C, may be accelerated when encapsulated within a liposome.

Objects of the invention:

An object of the present invention is to enhance bioavailability and to increase the dissolution rate in liposomal composition.

The present invention relates to a liposomal composition comprising an active ingredient, lipids as encapsulation material and a solvent, wherein the encapsulation material and solvent are present in a weight ratio of 1:1 to 1:5 and active ingredient and solvent are present in a weight ratio of 1 : 1 to 1 :5.

The active ingredients as used herein are curcumin, vitamin c or Coenzyme Q10. Other objects and benefits of the present invention will be more apparent from the following description, which is not intended to bind the scope of the present invention.

Summary of the invention:

Accordingly, a liposomal composition and a process for preparation thereof are disclosed.

The present invention helps in enhancing bioavailability and stability and dissolution rate in colloidal drug delivery systems.

The present invention relates to a liposomal composition comprising an active ingredient, lipids as encapsulation material and a solvent, wherein the encapsulation material and solvent are present in a weight ratio of 1:1 to 1:5 and active ingredient and solvent are present in a weight ratio of 1 : 1 to 1 :5.

The active ingredients as used herein are curcumin, vitamin c or Coenzyme Q10.

Typical routes of administering such pharmaceutical composition include, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal.

Typically, such a composition is in the form of a tablet or a capsule or a liquid preparation.

Detailed description of the invention:

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.

The embodiment of the present invention provides liposomal composition for enhancing bioavailability of the active ingreident. The effective carrier is used to enhance the stability, bioavailability and targeting property of active ingredient.

The present invention relates to a liposomal composition comprising an active ingredient, lipids as encapsulation material and a solvent, wherein the encapsulation material and solvent are present in a weight ratio of 1:1 to 1:5 and active ingredient and solvent are present in a weight ratio of 1 : 1 to 1 :5.

The terms used in the specification are defined as follows.

As used herein, the terms "comprising" (and any form of comprising, such as "comprise", "comprises", and "comprised"), "having" (and any form of having, such as "have" and "has"), "including" (and any form of including, such as "includes" and "include"), or "containing" (and any form of containing, such as "contains" and "contain"), are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

A "subject," "individual," or "patient," is used interchangeably herein, which refers to a vertebrate, preferably a mammal, more preferably a human. Tissues, cells and their progeny of a biological entity obtained in vitro or cultured in vitro are also encompassed.

The use of the terms "a" and "an" and "the" and similar referents in the context of describing the elements (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprising," "having," "including," and "containing" are to be construed as open-ended terms (i.e. , meaning "including, but not limited to,") unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the embodiments and does not pose a limitation on the scope of the claims unless otherwise stated. No language in the specification should be construed as indicating any non- claimed element as essential to the practice of the invention.

As used herein, the terms "treat," "treated," or "treating" mean both therapeutic treatment or prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder or disease, or obtain beneficial or desired clinical results. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of extent of condition, disorder or disease; stabilized (i.e. , not worsening) state of condition, disorder or disease; delay in onset or slowing of condition, disorder or disease progression; amelioration of the condition, disorder or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder or disease.

In another embodiment, a pharmaceutical composition is provided comprising an active ingredient, or a stereoisomer, or ester or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

The lipids include natural and/or synthetic phospholipids (Phosphatidylethanolamine, Phosphatidylglycerol, Phosphatidylcholline, Phosphatidylserine, Phosphatidylinositol) Phosphatidylcholine (also known as lecithin) and phosphatidylethanolamine constitute the two major structural components of most biological membranes. Liposome bilayers may also contain other constituents such as cholesterol, hydrophilic polymer conjugated lipids and water. The vehicles are selected from the group of Petrolatum, dimethyl sulfoxide and mineral oil.

In another embodiment, the present invention relates to a process for preparation of liposomal composition in colloidal drug delivery system comprising the steps of; a. Mixing lipid with a suitable solvent in a weight ratio of 1:3 in a beaker; b. Mixing active ingredient with suitable solvent in a weight ratio of 1:2 in another beaker; c. Sonicating the beakers in step ‘a’ and ‘b’; d. Adding gum acacia with solvent in third beaker to form a continuous phase; e. adding the lipid mixture in step ‘a’ to continuous phase in step ‘d’; f. adding the active ingredient mixture in step ‘b’ to the continuous phase in step ‘d’ slowly; g. Evaporation of the solvents. wherein the said active ingredient is vitamin-C or Curcumin or Coenzyme Q10 and is in solid dispersion form.

In another embodiment, primary emulsifiers are selected from the group consisting of; dextrin; sodium lauryl sulfate; Polyoxyethylene glycol; alkyl phenol ethers; Polyoxyethylene glycol sorbitan alkyl esters; Sorbitan alkyl esters and Block copolymers of polyethylene glycol and polypropylene glycol; Dioctyl sodium sulfosuccinate (DOSS); Perfluorooctanesulfonate (PFOS); Linear alkyl benzene sulfonates; Lignosulfonate, sodium stearate; Benzalkonium chloride (BAC), Cetylpyridinium chloride (CPC), Benzethonium chloride (BZT); Alkyltrimethylammonium salts: cetyl trimethylammonium bromide (CTAB) and cetyl trimethylammonium chloride (CTAC) and cocam idopropyl betaine. In another embodiment, secondary emulsifiers are selected from the group consisting of; alkyl, aryl esters Cetyl alcohol; glyceryl monosterate; methyl cellulose, stearic acid, erythritol.

Solvents used are selected from the group consisting of; water; acetone; alcohols, benzene, glycerin, triethyl amine, toluene, diethyl ether; ethyl propyl ether; ethyl acetate, ethylene glycol, acetic acid, acetic acid, acetonitrile, carbon tetrachloride and chloroform, PEG-400.

Fillers are selected from the group consisting of; ground calcium carbonate (GCC), precipitated calcium carbonate (PCC), kaolin and talc.

On reconstitution with or without sonication of above solid dispersion powder in water or suitable vehicle give a stable liposome size of upto 250 urn diameter. Solid dispersion powder shall be used as ingredient in tablet, capsule or liquid preparations.

The pharmaceutical compositions of the invention can be prepared combining an active pharmaceutical ingredient of the invention with an appropriate pharmaceutically acceptable carrier, diluent or excipient, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, injections, gels and microspheres.

In another embodiment, the present invention relates to administering “an effective amount of the composition of invention” to the subject suffering from said disease. Accordingly, active pharmaceutical ingredient and pharmaceutical compositions containing them may be administered using any amount, any form of pharmaceutical composition via any route of administration effective for treating the disease. Typical routes of administering such pharmaceutical compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal. Pharmaceutical compositions of the invention are formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. Compositions that will be administered to a subject or patient may take the form of one or more dosage units. The dosage forms can also be prepared as sustained, controlled, modified and immediate dosage forms.

The liposome compositions according to the invention can be prepared by any conventional methods

1. Mechanical dispersion methods like sonication, micro emulsification, membrane extrusion, dried reconstituted vesicles, freeze-thawed liposomes, freeze drying, lipid film hydration.

2. Solvent dispersion methods like ethanol injection, ether injection, double emulsion, reverse phase evaporation vesicles, stable pluri lamellar vesicles.

3. Detergent removal methods like dialysis, column chromatography, dilution and the like.

The accurate selection of liposome preparation method depends the physicochemical characteristics of the material to be entrapped, choice of liposomal ingredients, nature of the medium in which the lipid vesicles are to be dispersed, effective concentration of the entrapped substance, optimum size and shelf life of the vesicle, and batch-to-batch reproducibility.

The vitamin-C liposomal composition according to invention is used for the treatment of scurvy or any other indication or as a dietary supplement or as a food additive. The composition can be used as a food additive in bakery products or food products.

The curcumin liposomal composition according to invention is used wherein the composition is used for the treatment of wound healing or as an anti inflammatory agent. The Coenzyme Q10 liposomal composition according to invention is used for the treatment of heart failure or nerve pain or fibromyalgia or tissue damage or migraine or multiple sclerosis or heart arrack any other indication or as a dietary supplement or as a food additive.

The foregoing examples are illustrative embodiments and are merely exemplary. A person skilled in the art may make variations and modifications without deviating from the spirit and scope of the invention. All such modifications and variations are intended to be included within the scope of the claims.

Example 1: Formulation according to the invention:

The composition for vitamin-C solid dispersion powder for liposomal preparation contains vitamin-C, lipid, gum acacia and solvent. The process for preparation for the same as follows - a. Mixing lipid with a suitable solvent in a weight ratio of 1:3 in a beaker; b. Mixing vitamin-C with suitable solvent in a weight ratio of 1:2 in another beaker; c. Sonicating the beakers in step ‘a’ and ‘b’; d. Adding gum acacia with solvent in third beaker to form a continuous phase; e. adding the lipid mixture in step ‘a’ to continuous phase in step ‘d’; f. adding the vitamin-C mixture in step ‘b’ to the continuous phase in step ‘d’ slowly; g. Evaporation of the solvents.

Vitamin-C can be replaced with curcumin or Coenzyme Q10 in this example.