Description:

FIELD OF THE INVENTION

The present invention provides a pharmaceutical composition comprising pure extract of Euphorbia Prostrata substantially free of heavy metals and microbial contamination and wherein said extract has a controlled particle size and the composition comprising said Euphorbia Prostrata extract provides a release of not less than about 65% of the dry extract of Euphorbia Prostrata in first 10 minutes and not less than about 85% of dry extract of Euphorbia Prostrata after total dissolution study of about 30 minutes.

The present invention further provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of heavy metals selected from lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As).
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BACKGROUND OF THE INVENTION

Euphorbia Prostrata is a small, prostrate, annual green herb widely distributed globally and used as antihemorrhoidal, anti-inflammatory, analgesic, hypolipidemic, antidiabetic, antidirroheal, antiasthmatic and for various skin diseases. Among the various anorectal and colonic diseases, hemorrhoids occupy a prominent position and have been the subject of numerous clinical studies. Hemorrhoidal disease is characterized by bleeding, without any pain. Fresh blood spots occur immediately, on defecation. However, pain occurs when the hemorrhoids are secondarily infected, or complicated by thrombosis and anal fissures. There exist several procedures for the treatment of hemorrhoids. There are few prior art literature that discloses use of oral pharmaceutical compositions comprising dry extract of Euphorbia Prostrata. However, none of the prior art teaches about the heavy metals present in the extract and method of reducing the same.

In the present day scenario, the importance of herbal drugs is increasing due to their lesser side effects and acceptability to the majority of the population of third world countries. However, it is observed that some plants have the unique ability of accumulating toxic heavy metals from contaminated soil called phyto-extractor. Euphorbia Prostrata has an ability to accumulate heavy metals such as lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As) which are absorbed from the soil. Euphorbia Prostrata shows the highest capacity for accumulation of the mixture of these elements (588 µg g DW-1). Moreover, Euphorbia Prostrata tend to extract Hg with bio-accumulation coefficient more than 1. Although, Euphorbia Prostrata is an essential herb for the treatment of haemorrhoids, the extract used for the preparation of the pharmaceutical composition may contain high quantity of these heavy metals which leads to several side effects such as arsenic and lead poisoning.

Synthetic therapeutics is not only expensive but a number of adverse side effects are associated with them. Hence, plants are possible sources of novel structural entities that could provide effective treatment to the prevailing diseases.

Based on aforesaid, present invention is focussed to work on the contents of heavy metals and microbial contaminants present in the Euphorbia extract and specifically, present invention provides Euphorbia Prostrata dry extract with reduced quantities of the heavy metals and microbial contaminants, wherein the amount of heavy metals and microbial contaminants is less than minimum acceptable amount.

Further, present invention provides a composition of pure Euphorbia Prostrata dry extract which has a controlled particle size that provides improved dissolution profile of Euphorbia Prostrata at specific site of action.

OBJECT OF THE INVENTION

Main object of the present invention is to provide a pharmaceutical composition comprising substantially pure Euphorbia Prostrata extract, wherein said extract is substantially free of heavy metals and microbial contaminants.

Another object of the present invention is to provide a pharmaceutical composition comprising substantially pure Euphorbia Prostrata extract, wherein said extract has a controlled particle size that provides improved dissolution profile of Euphorbia Prostrata at specific site of action.

Another object of the present invention is to provide above said pharmaceutical composition for the treatment of anorectal disease and colonic diseases such as hemorrhoids, fissures, cracks, fistulas, abscesses, and inflammatory bowel disease.

SUMMARY OF THE INVENTION

In one aspect of the present invention, there is provided a process for preparing substantially pure Euphorbia Prostrata extract substantially free of heavy metals and microbial contaminants.

In another aspect, the present invention provides a pharmaceutical composition comprising above said substantially pure Euphorbia Prostrata extract.

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of heavy metals selected from lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As).

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of microbial contaminants selected from Escherichia coli, Salmonella, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterobacteria.

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein the particle size distribution of dry extract of Euphorbia Prostrata i.e. d10 is not more than 10 µm; d50 is not more than 100 µm and d90 is not more than 350 µm.

In another aspect, the present invention provides a pharmaceutical composition for the treatment of anorectal disease and colonic diseases such as haemorrhoids, fissures, cracks, fistulas, abscesses, and inflammatory bowel disease, wherein said composition comprises a dry extract of Euphorbia Prostrata substantially free of heavy metals and microbial contaminants and possess a controlled particle size of d10 is not more than 10 µm; d50 is not more than 100 µm and d90 is not more than 350 µm.

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said pharmaceutical composition is formulated as a solid dosage form having tapped bulk density of 0.45 g/ml to 1.5 g/ml which is maintained by 10-1400 taps.

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata containing flavonoidal constituents such as apigenin-7-glycoside, luteolin-7-glycoside, apigenin, luteolin and quercetin, and phenolic compounds that are therapeutically useful for treatment of anorectal and colonic diseases due to their haemostatic and astringent properties, wherein said phenolic compounds constitutes ellagic acid, gallic acid and tannins.

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of the Euphorbia Prostrata containing flavonoids and phenolic compounds, wherein total flavonoids are 2.5-15.0% by weight calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are 12-50% by weight calculated as gallic acid, along with atleast one pharmaceutically acceptable excipient; and wherein the pharmaceutical composition comprise of the extract of the plant Euphorbia Prostrata from about 0.1 % to about 99% by weight.

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of the Euphorbia Prostrata containing flavonoids and phenolic compounds, wherein total flavonoids are 3.0% to 8.5% by weight calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are 12.0% to 44.0% by weight calculated as gallic acid, along with atleast one pharmaceutically acceptable excipient; and wherein the pharmaceutical composition comprise of the extract of the plant Euphorbia Prostrata from about 0.1 % to about 99% by weight.

In another aspect, the present invention provides a pharmaceutical composition comprising dry extract of the Euphorbia Prostrata containing flavonoids and phenolic compounds, wherein total flavonoids are 2.5-15.0% by weight calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are 12-50% by weight calculated as gallic acid, along with atleast one pharmaceutically acceptable excipient; and wherein the particle size distribution of dry extract of Euphorbia Prostrata i.e. d10 is not more than 10 µm; d50 is not more than 100 µm and d90 is not more than 350 µm; and wherein said dry extract is substantially free of lead (Pb), cadmium (cd), mercury (Hg) and arsenic (As); and wherein said lead is less than about 5.0ppm,
said cadmium is less than about 0.2ppm, said mercury is less than about 0.1ppm, and said arsenic is less than about 1 ppm.
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DETAILED DESCRIPTION

Brief Description of Drawings:
Fig. 1, represents differential scanning colorimetry of dry extract of Euphorbia Prostrata
Fig. 2, represents X-Ray diffraction pattern of dry extract of Euphorbia Prostrata
Fig. 3, represents Thermogravimetric Analysis of dry extract of Euphorbia Prostrata

Euphorbia Prostrata dry extract is prepared from the extract taken from Euphorbia Prostrata plant that are collected from several areas where it grows like roadside weeds, or is collected from commercial green houses and even the areas including regions that are contaminated with heavy metals due to higher industrial activity. It is known that the plants that grow near to the places having higher contamination of heavy metals leads to high accumulation of such elements in the plant roots and various other parts. Present invention is a result of the work done in controlling the heavy metals concentration in dry extract of the Euphorbia Prostrata and hence, the composition of the present invention comprises substantially pure Euphorbia Prostrata dry extract which is extracted by a method that helps in reduction of heavy metals irrespective of the source and the place from which the plant part is collected. Herein, the present invention has dealt with the side effects such as lead or arsenic poisoning which may occur due to the continuous use of Euphorbia Prostrata dry extract, hence making the composition of the present invention as more safer drug composition.

Another effect achieved by the present invention is the improved dissolution profile of Euphorbia Prostrata extract which is achieved by controlling the particle size of the dry extract of Euphorbia Prostrata.

Accordingly in main embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of heavy metals and microbial contaminants.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata wherein said extract is substantially free of heavy metals selected from lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As).

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of microbial contaminants selected from Escherichia coli, Salmonella, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterobacteria.

The Euphorbia Prostrata is found to have high tendency to accumulate arsenic and lead metals above the permissible level. As per Environ Sci Pollut Res (2013) 20:3946–3955, Blenda et al, it is analysed that the Arsenic and metals concentrations found in the Euphorbia Prostrata showed the highest capacity to accumulate arsenic as compared to other plants. Euphorbia Prostrata, is also found to be an indigenous plant that accumulated more lead (Pb) wherein it showed a stem>leaves capacity for Pb accumulation. Herein, present invention provides substantially pure Euphorbia Prostrata dry extract extracted from contaminated plant parts by a specific extraction method that results into dry extract having negligible to non-detectable amount of heavy metals.

Hence, in one embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of heavy metals selected from lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As), and wherein said lead is less than about 5.0ppm, preferably less than 3.0ppm, and preferably less than 1.0 ppm and most preferably below detection limit; and
wherein said cadmium is less than about 0.2ppm, preferably less than 0.10ppm, preferably less than 0.050ppm and most preferably below detection limit; and
wherein said mercury is less than about 0.1ppm, preferably less than 0.05ppm and most preferably below detection limit; and
wherein said arsenic is less than about 1 ppm, preferably less than 0.5ppm and most preferably are below detection limit.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of heavy metals selected from lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As), and wherein said heavy metals are below detection limit.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of microbial contaminants selected from Escherichia coli, Salmonella, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterobacteria; and
wherein said Enterobacteria is less than 10 per gram of the extract and wherein said Escherichia coli, Salmonella, Pseudomonas aeruginosa, and Staphylococcus aureus are not present in the extract.

In another embodiment, said microbial contaminants may further comprises of aerobic microbial contaminants, yeast and moulds.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of aerobic microbial contaminant wherein said aerobic microbial contaminants are less than about 30,000 cfu/g of the extract, preferably less than about 3,000 cfu/g of the extract and most preferably less than about 100 cfu/g of the extract, and most preferably 30 cfu/g of the extract.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of yeast and mould contaminants wherein said yeast and mould are in total less than about 1,000 cfu/g of the extract, preferably less than about 500 cfu/g of the extract and most preferably less than about 100 cfu/g of the extract, and most preferably less than about 10 cfu/g of the extract.

Moreover, present invention provides a composition that has improved dissolution profile of the dry extract of Euphorbia Prostrata at the specific site of action which is achieved by controlled particle size of said dry extract.

Accordingly, in one more embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein the particle size distribution of dry extract of Euphorbia Prostrata i.e. d10 is not more than 10 µm; d50 is not more than 100 µm and d90 is not more than 350 µm.

In a preferred embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein the particle size distribution of dry extract of Euphorbia Prostrata i.e. d10 is in the range of 3.5 µm to 10 µm; d50 is in the range of 30 µm to 80 µm and d90 is in the range of 150 µm to 350 µm.

In most preferred embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein the particle size distribution of dry extract of Euphorbia Prostrata i.e. d10 is in the range of 5.5 µm to 10 µm; d50 is in the range of 45 µm to 70 µm and d90 is in the range of 200 µm to 300 µm.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata having a controlled particle size, wherein the particle size distribution of dry extract of Euphorbia Prostrata i.e. d10 is not more than 10 µm; d50 is not more than 100 µm and d90 is not more than 350 µm, and said dry extract provides a release of not less than about 65% of the dry extract of Euphorbia Prostrata in first 10 minutes and not less than about 85% of dry extract of Euphorbia Prostrata after total dissolution study of about 30 minutes.

In another embodiment, the compositions of the present invention provides effective and improved dissolution profile as mentioned above when tested at physiological pH range of about 2 to about 7.5.

In another embodiment, the present invention provides a pharmaceutical composition for the treatment of anorectal disease and colonic diseases such as haemorrhoids, fissures, cracks, fistulas, abscesses, and inflammatory bowel disease, wherein said composition comprises a dry extract of Euphorbia Prostrata substantially free of heavy metals and microbial contaminants and possess a controlled particle size of d10 is not more than 10 µm; d50 is not more than 100 µm and d90 is not more than 350 µm.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata containing flavonoidal constituents such as apigenin-7-glycoside, luteolin-7-glycoside, apigenin, luteolin and quercetin, and phenolic compounds that are therapeutically useful for treatment of anorectal and colonic diseases due to their haemostatic and astringent properties, wherein said phenolic compounds constitutes ellagic acid, gallic acid and tannins.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of the Euphorbia Prostrata containing flavonoids and phenolic compounds, wherein total flavonoids are 2.5-15.0% by weight calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are 12-50% by weight calculated as gallic acid, along with atleast one pharmaceutically acceptable excipient; and wherein the pharmaceutical composition comprise of the extract of the plant Euphorbia Prostrata from about 0.1 % to about 99% by weight.

In preferred embodiment, the present invention provides a pharmaceutical composition comprising dry extract of the Euphorbia Prostrata containing flavonoids and phenolic compounds, wherein total flavonoids are 3.0% to 8.5% by weight calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are 12.0% to 44.0% by weight calculated as gallic acid, along with atleast one pharmaceutically acceptable excipient; and wherein the pharmaceutical composition comprise of the extract of the plant Euphorbia Prostrata from about 0.1 % to about 99% by weight.

In most preferred embodiment, the present invention provides a pharmaceutical composition comprising dry extract of the Euphorbia Prostrata containing flavonoids and phenolic compounds, wherein total flavonoids are about 4.2% w/w calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are about 20.4% w/w % calculated as gallic acid, along with atleast one pharmaceutically acceptable excipient; and wherein the pharmaceutical composition.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of the Euphorbia Prostrata containing flavonoids and phenolic compounds, wherein total flavonoids are 2.5-15.0% by weight calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are 12-50% by weight calculated as gallic acid, along with atleast one pharmaceutically acceptable excipient; and wherein the particle size distribution of dry extract of Euphorbia Prostrata i.e. d10 is not more than 10 µm; d50 is not more than 100 µm and d90 is not more than 350 µm; and wherein said extract is substantially free of lead (Pb), cadmium (cd), mercury (Hg) and arsenic (As); and wherein said lead is less than about 5.0ppm, said cadmium is less than about 0.2ppm, said mercury is less than about 0.1ppm, and said arsenic is less than about 1 ppm.

Moreover, in one more embodiment, the dry extract of the Euphorbia Prostrata used in the compositions of the present invention are characterized by differential scanning colorimetry with onset peak at 51.12oC and having an endothermic peak at 56.24oC.

In another embodiment, the dry extract of the Euphorbia Prostrata used in the compositions of the present invention are characterized by Powder X-Ray diffraction pattern at 26.62, 28.17 ± 2o 2? with water content of less than about 4.0%, and preferably the water content is less than about 2.5%, most preferably 1.5% or less.

One important characteristic is tapped bulk density, wherein the maximum packing density of a powder (or blend of powders) is achieved under the influence of well defined, externally applied forces. The packed volume or density depends on a number of factors including particle size distribution, particle shape and cohesiveness due to surface forces including moisture. Greater Tap density requires that loose powders be compacted into a durable solid form using external mechanical force which is formulated as tablets or filled in capsules. The present invention provides a composition of extract of Euphorbia Prostrata, wherein the particle size and moisture content of said extract is maintained at a level as described above, that leads to the optimum flowability of the dry powder which when tapped provides tapped density ranging between 0.45 g/ml to 1.5 g/ml.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said pharmaceutical composition is formulated as a solid dosage form having tapped bulk density of 0.45 g/ml to 1.5 g/ml which is maintained by 10-1400 taps.

In a preferred embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said pharmaceutical composition is formulated as a solid dosage form having tapped bulk density of 0.68 g/ml after 10 taps, 0.81 g/ml after 50 taps and 0.83 g/ml after 1250 taps.

The pharmaceutical composition of the present invention can be formulated into different dosage forms using pharmaceutically acceptable excipient(s) and techniques known to art. Pharmaceutical dosage forms of the present invention can be in the form selected from but not limited to, tablets (coated or uncoated), chewable tablets, mini-tablets, troches and lozenges, sachet, capsules (hard or soft), mini-tablets/tablets filled in capsule, granules, creams, solutions, suspensions, powder, sublingual dosage forms, wafers, caplets, and other dosage forms suitable for oral administration.

In an embodiment, the compositions can be prepared by dissolving or dispersing dry extract of Euphorbia Prostrata in a solvent or mixture of solvents with or without pharmaceutically acceptable excipients and depositing or layering or spraying the solution or dispersion onto inert beads, spheres, cores, seeds, particles or nuclei.

In a preferred embodiment of the present invention, the composition is formulated in the form of tablets. The tablets can be prepared by either direct compression, dry compression (slugging) or by wet granulation. In a preferred embodiment of the present invention, the oral composition is prepared by direct compression/compaction, slugging, extrusion, moulding, or the like using excipients known to the person skilled in the art. The composition may also be prepared by wet granulation technique, which may use either aqueous/polar solvent(s) or non-aqueous/organic solvent(s) or mixtures thereof.

In another embodiment, the coated tablets/mini-tablets may optionally comprise a part or whole of the active agent in the coating composition.

In an embodiment of the present invention, the non-aqueous solvent used is selected from but not limited to a group comprising ethanol, isopropyl alcohol, methylene chloride or acetone.

In an embodiment of the present invention, a suitable organic solvent system is selected from but not limited to, methanol, ethanol, 1-butanol, 2-butanol, 3-methyl-1-butanol, 1-propanol, 2-propanol, isopropanol, 1-pentanol, acetone, methyl acetate, ethyl acetate, butyl acetate, propyl acetate, isopropyl acetate, isobutyl acetate, ethyl- ether, tert-butylmethyl ether, ethyl formate, chloroform, dichloromethane, and the like or mixtures thereof.

In a further embodiment, the creams are prepared by technique which may use the method of dispersing dry extract in non-aqueous solvent such as alcohols selected from ethanol, isopropyl alcohol and the like.

In a further embodiment, the pharmaceutical composition of the present invention further comprises pharmaceutically acceptable excipients selected from but limited to, the group comprising of diluents, disintegrants, binders, anti-adherants, glidants, opacifier, buffering agents, colorants, flavoring agents, coating agents, solvents, viscosifying agents, waxes, wetting agents, emulsifying agents, solubilizers, stabilizers, buffering agents, fillers, bulking agent, thickeners such as sorbitol, organic acids, colorants, stabilizers, preservatives, lubricants, chelating agents, pH adjusting agents, antioxidants, humectants, osmotic agents, and mixture thereof.

The diluents used in the present invention is selected from lactose, mannitol, sorbitol, starch, microcrystalline cellulose, xylitol, fructose, sucrose, dextrose, sorbitan stearate, dicalcium phosphate, calcium sulphate and the like.

The solubility enhancing agents or solubilizers used in the present invention is selected from glycerin, various grades of polyethylene oxides, transcutol, glycofurol, polysorbate, and the like.

The disintegrants used in the present invention includes but not limited to starch or its derivatives, fully pregelatinized maize starch (Starch 1500®), corn oil, croscarmellose sodium, sodium starch glycollate, crosspovidone and the like.

The binders used in the present invention is selected from but not limited to a group comprising polyvinyl pyrrolidone, starch or its derivatives, fully pregelatinized starch (USNF+1H), hydroxypropyl methyl cellulose, corn oil, hydroxypropyl cellulose, gum arabic powder, gelatin and the like used alone or in combination thereof.

The glidants used in the present invention is selected from but not limited to a group comprising of corn starch, colloidal silicon dioxide, silica derivatives, pyogenic silica, talc, ascorbyl palmitate, calcium palmitate, magnesium stearate, and mixture thereof.

The lubricants used in the present invention includes but not limited to talc, magnesium stearate, calcium stearate, stearic acid, hydrogenated vegetable oil, sodium starch fumarate, glyceryl behenate, glycerine, and the like used either alone or in combination thereof.

The stabilizers used in the present invention includes but not limited to glyceryl behenate, sodium carboxymethyl cellulose, gelatin, pectin, sodium alginate, guar gum, povidone, hydropropyl methyl cellulose, hydroxypropyl cellulose, sodium pyrophosphate, carrageenan, locust bean gum, veegum, polysorbate, and the like used either alone or in combination thereof.

The preservatives used in the present invention includes parabens such as methyl parabens, propyl parabens, glycol ethers, diazolidinyl urea, butylated hydroxyanisole, glycerine, and the like.

The antioxidants used in the present invention includes butylated hydroxyanisole, propyl gallate, sodium metabisulfite, vitamin C, Vitamin E, lutein, beta-carotene, citric acid, ascorbic acid, and the like.

The emulsifying agent used in the present invention includes glycerine, glyceryl monostearate, polawax, lecithin, sodium lauryl sulfate, behentrimonium methosulphate, bees wax, Candelilla wax, veegum, propylene glycol, polysorbate and the like.

The humectants and viscosity decreasing agents used in the present invention includes propylene glycol, glycerine, other glycols such as butylene glycol, triethylene glycol, collagen and glycerol.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata wherein said composition further comprises of one or more of other pharmaceutically active agent selected from vasoprotective agent or angioprotective agents, anesthetics, vasoconstrictors, other protectants, counterirritants, astringents, wound healing agents, antimicrobial agents, keratolytics, anticholinergics and mixture thereof;
wherein vasoprotective agents or angioprotective agents are selected from calcium dobesilate, monoethanolamine oleate, polidocanol, heparin, sodium apolate, tribenoside and wherein the amount of vasoprotective agents or angioprotective agents could vary from 25% to 65% by weight; and
wherein anesthetics are selected from benzocaine, diperodon, pramoxine, camphor, dibicaine, phenol, tetracaine, lignocaine, phenacaine and wherein the amount of anesthetics could vary from 0.25% to 25% by weight; and
wherein vasoconstrictors are selected from ephedrine and phenylephedrine and the amount of vasoconstrictors could vary from 0.005% to 1.5% by weight; and
wherein other protectants are selected from aluminium hydroxide gel, calamine, cocoa butter, cod or shark liver oil, glycerine, kaolin, lanolin, mineral oil, starch, white petroleum, wool alcohol, zinc oxide, vegetable or castor oil, polyethylene glycol, and propylene glycol and wherein the amount of other protectants could vary from 5.0% to 88.0% by weight; and
wherein counterirritants is menthol in aqueous solution in the range of 0.25% to 2.5% by weight; and
wherein astringents are selected from the group comprising of calamine, zinc oxide, hemamelis water, bismuthresorcinol compound, bismuth subgallate, Peruvian balsam, aluminium chlorhydroxy allantoinate, tannic acid and tannins, wherein the amount of astringents could vary from 0.2% to 60% by weight; and
wherein wound healing agents are vitamin A and Vitamin D ranging between 0.005% and 0.04% by weight; and
wherein antimicrobial agents are selected from the group comprising of benzethonium chloride, benzalkonium chloride, boric acid, 8-quinolinol benzoate, secondary amyltricresols, cetylpyridinium chloride, phenol, methanol, chlorothymol, camphor, and wherein the amount of antimicrobial agents is between 0.02% and 40.0% by weight; and
wherein keratolytics are selected from aluminium chlorhydroxy allantoinate and resorcinol and the amount of keratolytics could vary from 0.2% to 3.5% by weight; and
wherein anticholinergics are selected from atropine, solanaceous alkaloids and wherein the amount of anticholinergics could vary from 0.02% and 0.1% by weight.

In preferred embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata wherein said composition further comprises of vasoprotective agent or angioprotective agents such as calcium dobesilate in an amount of 25% to 65% by weight; and wherein the pharmaceutical composition comprises the extract of the plant Euphorbia Prostrata from about 0.1 % to about 99% by weight.

In another embodiment, the formulation is provided in forms of tablets or capsules comprising 25-300mg of the dry extract of Euphorbia Prostrata, preferably, 50-250mg and most preferably 100mg along with the excipients wherein the excipients are in the amount ranging from 50% to 90% by weight, preferably 65% to 85% by weight of the tablet; and wherein the tablet is preferably coated by Opadry.

In another embodiment, the formulation is provided in forms of creams or ointments comprising 60-90% v/v of alcoholic extract of Euphorbia Prostrata present in an amount of 0.1% to 15% w/w, preferably 5-10% w/w along with cream base.

In another embodiment, the present invention provides a pharmaceutical composition comprising dry extract of Euphorbia Prostrata substantially free of heavy metals and microbial contaminants, wherein said extract is prepared by a process comprising the steps of:
a) drying Euphorbia Prostrata and grounding to coarse powder;
b) extracting in polar solvent selected from alcohols and ketones to get an extract, wherein the polar solvent is added with or without water;
c) concentrating the extract and washing with non-polar solvent selected from hydrocarbon or chlorinated hydrocarbon;
d) extracting the washed extract with ester or ketone solvent;
e) optionally dehydrating the extract of step d); and
f) drying and milling to powder form.

In another embodiment, the polar solvent is selected from ethanol, methanol, tert-butanol, isopropyl alcohol, acetone, and methyl ethyl ketone.

In another embodiment, the ester solvent is preferably, ethyl acetate and ketone solvent used in step d) is preferably methyl ethyl ketone.

In another embodiment, the present invention provides dry extract of Euphorbia Prostrata, wherein said dry extract is substantially free of residual solvents, and wherein the residual solvents such as ethanol is less than about 5000ppm, and preferably less than about 500ppm, and most preferably less than about 210ppm;
and wherein ethyl acetate is less than about 5000ppm, and preferably less than about 500ppm, and most preferably less than about 300ppm;
and wherein hexane is less than about 290ppm, and preferably less than about 100ppm, and most preferably less than about 10ppm, and less than about 6.5ppm.

EXAMPLES

The examples given below serve to illustrate embodiments of the present invention. However they do not intend to limit the scope of present invention in any manner whatsoever.

Example 1: Preparation of dry extract of Euphorbia Prostrata

The powdered Euphorbia Prostrata was packed in a S.S. extractor. The extraction was performed using 80% aqueous methanol at about 60°C till the plant was totally exhausted. The aqueous-methanolic extracts so obtained were combined and concentrated by distillation. The concentrated extract was washed with 5-10 volumes of hexane to remove the wax and fatty material. The washed extract was dried completely and then extracted using ethyl acetate. The ethyl acetate extract was dehydrated with anhydrous sodium sulphate and concentrated by distillation. The concentrated extract was dried completely for several hours at 60°C under vacuum. The final purified extract was milled to a fine powder, sifted for uniform particle size and packed to protect from the moisture.

Example 2: Tablet composition of dry extract of Euphorbia Prostrata

Ingredients Amount (in mg)
Euphorbia Prostrata extract 100
Mannitol IP 348.85
Microcrystalline cellulose IP+IH 41.00
Pregelatinized starch 5.50
Purified water Lost in processing
Croscarmellose sodium 10.00
Glyceryl Behenate 3.00
Colloidal silicon dioxide 3.00
Magnesium stearate IP+IH 1.65
Crospovidone 37.00
Coating-I
Opadry II (white) 13.75
Purified water Lost in processing
Coating-II
Opadry (Green) 19.73
Purified water Lost in processing

Process:
i) Dry extract of Euphorbia Prostrata, microcrystalline cellulose and mannitol were sifted from sieve of mesh size #30 and mixed together;
ii) Pregelatinized starch was dispersed in purified water with continuous stirring to prepare the binder solution;
iii) The binder solution of step (ii) was mixed with the material of step (i) to obtain the granules followed by drying of the granules;
iv) Croscarmellose sodium, glyceryl behenate, colloidal silicon anhydrous, magnesium stearate, crospovidone were sifted together through a sieve of mesh size #30;
v) The dried granules of step (iii) and the material of step (iv) was blended together for 10-12 minutes;
vi) The blended granules of step (v) were compressed into tablets;
vii) Opadry II was mixed in purified water for about 45 minutes to prepare the base coating solution;
viii) The core tablets of step (vi) were coated with the base coating solution obtained from step (vii);
ix) Opadry (Green) was mixed in purified water for about 45 minutes to prepare the final coating; and
x) The base coated tablet of step (viii) were coated with final coating solution obtained from step (ix).

Example 3: Tablet composition of dry extract of Euphorbia Prostrata and Calcium Dobesilate Monohydrate

Ingredients Amount (in mg)
Euphorbia Prostrata extract 100
Clacium Dobesilate Monohydrate 500
Starch (Maize) 50
Microcrystalline cellulose IP+IH 65.5
Croscarmellose sodium 30.00
Crospovidone 30.00
Sodium metabisufite 0.50
Purified water Lost in processing
Croscarmellose sodium IP 20.00
Crospovidone IP+IH 40.00
Maize Starch IP 50.00
Colloidal Silicon dioxide 5.00
Magnesium stearate 9.00
Film Coating
200 series (white) 22.50
Purified water Lost in processing
Coating-II
Opadry 200 (Green) 23.00
Purified water Lost in processing

Process:
i) Dry extract of Euphorbia Prostrata, Calcium Dobesilate Monohydrate, starch, microcrystalline cellulose, croscarmellose sodium, crospovidone were sifted from sieve of mesh size #40 and mixed together and blended for 15 min;
ii) Sodium metabisulfite was dispersed in purified water with continuous stirring to prepare the binder solution;
iii) The binder solution of step (ii) was mixed with the material of step (i) to obtain the granules followed by drying of the granules;
iv) Croscarmellose sodium, crospovidone and starch were sifted together through a sieve of mesh size #40;
v) The dried granules of step (iii) and the material of step (iv) was blended together for 10-12 minutes;
vi) Magnesium stearate and colloidal silicon dioxide were sifted together through a sieve of mesh size #60, mixed and blended with the blend of step (v);
vii) The blended granules of step (vi) were compressed into tablets;
viii) 200 series was mixed in purified water for about 45 minutes to prepare the film coating solution and coated the tablets of step (vii);
ix) Opadry (Green) was mixed in purified water for about 45 minutes to prepare the film coating dispersion solution; and
x) The film coated tablet of step (viii) were coated with film coating dispersion solution obtained from step (ix) and dried.

Example 4: Cream composition of ethanolic 80% v/v extract of Euphorbia Prostrata

Ingredients Amount (in mg)
Euphorbia Prostrata dry extract ethanolic 80% v/v [(35-70):1] IH 10
Propylene Glycol IP 50.00
Methyl paraben IP 1.50
Propyl Paraben IP 0.30
Titanium dioxide 10.00
Emulsifying wax 170.00
Sorbitan Stearate 20.00
Corn oil 100.00
Butylated Hydroxyanisole 0.029
Propyl Gallate IP 0.029
Glycerine IP 50.00
Sorbitol solution (70% non-crystallizing) IP 30.00
Veegum HV 10.50
Sodium carboxymethyl cellulose 3.15
Polysorbate 80 15.00
Purified water q.s

Process:
Preparation of oily phase:
i) Filtered corn oil through nylon cloth of mesh size #160;
ii) Emulsifying wax, sorbitan stearate, butylated hydrxyanisole, propyl gallate were mixed with corn oil of step (i), followed by heating at 70-75oC.

Preparation of aqueous phase:
iii) Veegum HV was sifted through sieve of mesh size #40 and dispersed in purified water under continuous stirring;
iv) Sorbitol solution and polysorbate were filtered through nylon cloth of mesh size #160 and mixed with step (iii) under continuous stirring;
v) Sifted sodium carboxymethyl cellulose through sieve of mesh size #20, filtered glycerine through nylon cloth of mesh size #160;
vi) Mixed filtered glyceine with sifted sodium carboxymethyl cellulose followed by mixing with step (iv) under continuous stirring maintaining the quantity with purified water to obtain the bulk;
vii) Maintained the temperature of the bulk to 70-75oC.

Preparation of Euphorbia Prostrata dry extract ethanolic 80% (v/v) [35-70):1] solution:
viii) Filtered propylene glycol through nylon cloth of mesh size #160 and heated to temperature of 60-65oC;
ix) Sifted Euphorbia Prostrata dry extract ethanolic 80% (v/v) [35-70):1], propyl paraben, methyl paraben through sieve of mesh size #40 and sifted titanium dioxide through sieve of mesh size #100;
x) Mixed sifted material with filtered propylene glycol.

Preparation of cream:
xi) Mixed the oily phase and aqueous phase together which are maintained at a temperature of 70-75oC, and cooled the emulsion so obtained to 50-55oC; and
xii) Added propylene glycol and homogenized the bulk for 10 minutes, and cooled the emulsion (cream) so obtained to 20-25oC, followed by packing.

Dissolution Study Method
The dissolution study method of the present invention has the following parameters:
Dissolution medium : 0.5% w/v sodium lauryl sulphate in distilled water
Dissolution medium volume : 1000 ml Apparatus : Paddle (USP Type II)
Paddle Speed : 50 rpm
Dissolution time •. 10, 30 minutes

The dissolution profile of the composition of the present invention is as follows:
Example Time (minutes) Dissolution profile (% drug release)
Example 2 10 70%
15 78%
20 80%
25 82%
30 85.5%
Example 3 10 72%
15 77%
20 79%
25 81%
30 85.0%
Example 4 10 75%
15 79%
20 80%
25 85%
30 88%

The Detailed elemental analysis of the Euphorbia Prostrata is as follows:
Element (Heavy Metal) Result Units
Li 2.67 ppm
V 1.69 ppm
Cr 2.87 ppm
Co 0.722 ppm
Ni 2.94 ppm
Cu 28.81 ppm
As 0.42 ppm
Se 0.366 ppm
Mo 0.107 ppm
Ru ND ppm
Rh 0.012 ppm
Pd 0.004 ppm
Ag ND ppm
Cd 0.033 ppm
Sn ND ppm
Sb 0.039 ppm
Os 0.002 ppm
Ir 0.032 ppm
Pt 0.001 ppm
Au 0.079 ppm
Hg ND ppm
Tl 0.020 ppm
Pb 0.981 ppm , Claims:WE CLAIM

1. A pharmaceutical composition comprising dry extract of Euphorbia Prostrata, wherein said extract is substantially free of heavy metals and microbial contaminants.

2. The composition as claimed in claim 1, wherein said heavy metals are selected from lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As).

3. The composition as claimed in claim 2, wherein said lead is less than about 5.0ppm, the cadmium is less than about 0.2ppm, the mercury is less than about 0.1ppm and the arsenic is less than about 1 ppm.

4. The composition as claimed in claims 1-3, wherein said heavy metals are below detection limit.

5. The composition as claimed in claim 1, wherein said microbial contaminants includes Escherichia coli, Salmonella, Pseudomonas aeruginosa, Staphylococcus aureus, Enterobacteria, aerobic microbial contaminants, yeast and moulds; and wherein said enterobacteria is less than 10 per gram of the extract, the aerobic microbial contaminants are less than about 30,000 cfu/g of the extract, and the yeast and mould are in total less than about 1,000 cfu/g of the extract.

6. The composition as claimed in claim 1, wherein said dry extract of Euphorbia Prostrata has a controlled particle size distribution with d10 not more than 10 µm; d50 not more than 100 µm and d90 not more than 350 µm.

7. A pharmaceutical composition for the treatment of anorectal disease and colonic diseases such as hemorrhoids, fissures, cracks, fistulas, abscesses, and inflammatory bowel disease, wherein said composition comprises a dry extract of Euphorbia Prostrata substantially free of heavy metals and microbial contaminants and having a controlled particle size distribution of d10 not more than 10 µm; d50 not more than 100 µm and d90 not more than 350 µm,
wherein dry extract of Euphorbia Prostrata containing flavonoidal constituents including apigenin-7-glycoside, luteolin-7-glycoside, apigenin, luteolin and quercetin, and phenolic compounds including ellagic acid, gallic acid and tannins; and
wherein total flavonoids are 2.5-15.0% by weight calculated as apigenin-7-glycoside; and wherein the total phenolic compounds are 12-50% by weight calculated as gallic acid,
along with atleast one pharmaceutically acceptable excipient, optionally one or more of other pharmaceutically active agent; and
wherein the pharmaceutical composition comprise of the extract of the plant Euphorbia Prostrata from about 0.1 % to about 99% by weight.

8. The composition as claimed in claim 7, wherein said other pharmaceutically active agent is selected from vasoprotective agent or angioprotective agents, anesthetics, vasoconstrictors, other protectants, counterirritants, astringents, wound healing agents, antimicrobial agents, keratolytics, anticholinergics and mixture thereof.

9. The composition as claimed in claims 7 and 8, wherein said other pharmaceutically active agent is Calcium Dobesilate.

10. The composition as claimed in claim 7, wherein said composition provides a release of not less than about 65% of the dry extract of Euphorbia Prostrata in first 10 minutes and not less than about 85% of dry extract of Euphorbia Prostrata after total dissolution study of about 30 minutes.

Dated this, 5th Day of Apr, 2023 For Mankind Pharma Ltd.

Dr. Anil Kumar
Chief Scientific Officer