Title of Invention	"PROCESS FOR THE PREPARATIN OF A NITRO-GROUP BASED TOPICAL FORMULATED FOR TREATMENT OF ACUTE PERIPHERAL MICROVASCULAR DISEASEAS."
Abstract	The present invention relates to a process for the preparation of a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. The present invention also relates to a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. More particularly, the present invention relates to a nitro group based topical formulation for the treatment of cold injuries and other peripheral vascular diseases such as those afflicting microvasculature or microcirculation.

Title of Invention

"PROCESS FOR THE PREPARATIN OF A NITRO-GROUP BASED TOPICAL FORMULATED FOR TREATMENT OF ACUTE PERIPHERAL MICROVASCULAR DISEASEAS."

Abstract

The present invention relates to a process for the preparation of a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. The present invention also relates to a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. More particularly, the present invention relates to a nitro group based topical formulation for the treatment of cold injuries and other peripheral vascular diseases such as those afflicting microvasculature or microcirculation.

Full Text	PROCESS FOR THE PREPARATION OF A NITRO-GROUP BASED TOPICAL FORMULATION FOR TREATMENT OF ACUTE PERIPHERAL MICROVASCULAR DISEASES FIELD OF THE INVENTION The present invention relates to a process for the preparation of a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. The present invention also relates to a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. More particularly, the present invention relates to a nitro group based topical formulation for the treatment of cold injuries and other peripheral vascular diseases such as those afflicting microvasculature or microcirculation. BACKGROUND OF THE INVENTION Vasospastic/vasodestructive or ischemic disorders of limbs are very common. These are found even in apparently normal individuals, and generally occur due to transient or permanent reduction of blood supply and oxygen at tissue level. Such disorders occur more frequently and severely in cold injuries, diabetes mellitus, Berger's disease, Raynaud's phenomenon, burns, arteritis of all types and other such diseases coming under the broad category of peripheral vascular diseases, particularly involving microcirculation or microvasculature. Hypovolemia, atherosclerosis, anemia and old age are further risk factors. Thrombosis in microvasculature of hands or feet causes additional complications, leading to further reduction in blood and oxygen supply and permanent damage of the tissues. Such disorders can lead to tissue death, gangrene and loss of limb or function. Such conditions are further aggravated in cold weather, particularly at high altitude and in old individuals and those who consume tobacco products or are exposed to second hand effects of tobacco products. Another related problem or complication is that local ischemia leads to development of infection and inflammation that further causes more local ischemia resulting in a vicious cycle. These peripheral vascular diseases could be of different pathological origins, namely autoimmune, infective, environmental, toxic, autonomic, vascular, idiopathic and of other and unknown origins. The disorders as described above lead to acute pain, cyanosis and loss of work efficiency (particularly of arms and legs, hands and fingers, feet and toes or facial parts) leading to edema and ulcer formation, leading to muscle and bone death, deep-seated infection, amputation and ultimately loss of the affected body organ. Even when the local blood and oxygen supply is partially restored by conventional treatment, the tissue injury aggravates due to oxidative mechanisms. Barring very mild injuries, the damage at microvascular or micro-circulation level is permanent and may be progressive. Most of the predisposing or precipitating factors are either accidental or unpreventable. It is known in the art to treat peripheral vascular diseases, particularly cold injuries in a non-pharmacological manner such as by slow re-warming of the affected part in water, called thawing. A practical limitation with of the above method is that this cannot be used as a preventive routinely but can only be used as to manage the condition. Another drawback is that it is neither practical nor possible to use thawing as the main course of treatment in field conditions. Another limitation of the slow re-warming is that the method is useful only for cold injuries and not other forms of peripheral vasospastic/ ischemic diseases. Even in cold injuries, thawing is useful for only mild cold injuries and is inadequate in major cold injuries such as frostbite. Thawing is also a very slow method of treatment and does not provide speedy relief Various pharmacological methods are also known in the art for treatment of ischemic or vasospastic/vasodestructive disorders. Such pharmacological methods include systemic methods such as oral, intramuscular or intravenous administration of vasodilator drugs. However, such pharmacological methods suffer from several limitations. A major limitation of the above method is that there is disagreement about its benefit in treating local micro-vascular conditions (mostly of arms and legs or hands and feet). This is primarily due to the inability of the systemic route to deliver sufficient quantity of medication to the peripheral ischemic lesion because of large volume of distribution. One of the most potent and fast-acting vasodilators, nitroglycerine or amylnitrate, if given by systemic route for local action on the aforesaid lesions, would result in very high pharmacological dose in the blood and is therefore impractical. Another significant disadvantage of pharmacological methods are that there may not be any benefit at all if blood supply to the hand/foot is already blocked significantly due to thrombosis. Additionally, the dosage required for systemic route is high and can cause serious side effects such as low blood pressure and significant palpitation and heart problems. This limits the duration of treatment to a few days, thereby limiting the benefits of such treatment, if any. Thus, this route cannot deliver potent and fast-acting vasodilators such as nitroglycerine, even though these would be quite effective for lesion treatment. This method also results in high treatment cost due to the high dosages required for managing the disease. Another method known in the art for management of the above diseases comprise systemic administration of anti-coagulants and/or thrombolytic agents. However, similar to systemic administration of vasodilators, the above method of management has the drawback that it requires a very high dosage thereby increasing side effects (bleeding from unrelated sites) and cost. The use of anti-coagulants and thrombolytic drugs are also considered dangerous except in hospital conditions. Another drawback is that these agents cannot be used in chronic or recurring conditions. Their use at present is therefore limited to very severe cases considering the risks involved. Another method known in the art for management of the above diseases is systemic administration of antiplatelet drugs Uke dipyridamole or aspirin. Similar to systemic administration of vasodilators, the above method of management has the drawback that it requires a very high dosage thereby increasing side effects and cost. Another drawback of the above method is that the pharmacological effect of such drugs is limited if the local blood circulation is compromised due to thrombosis. It is also known in the art to manage the above diseases using oral pentoxyphyllin or other Xanthine-based drugs or Dextran infusion that are known to reduce blood viscosity, thus enabling more flow of blood to the lesions. However, the above method, like systemic vasodilators and antiplatelet drugs, has the drawback that a very high blood concentration is required for optimal concentration in the peripheral lesions, thus increasing side effects and cost. Also, it may have limited pharmacological effect if local blood circulation is compromised due to thrombosis. Topical ointments containing analgesics and antibiotics are available but have the drawback that these are useful in controlling only the infection and do not address the main problem of microvascular insufficiency. Topical ointments containing nicotinic acid as a vasodilator (for example, 'ThrombophobeTM) are available in the market for treating microvascular insufficiency. Their main drawback is that though useful, these are usually not found sufficiently potent to cure the conditions by themselves and are only used for supportive treatment. As such, there is need of a topical ointment which could penetrate effectively through the skin to deliver fast-acting and potent drugs that act on legal microvasculature and increase the local blood flow through vasodilation and other effects, OBJECTS OF THE EVVENTION The main object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process for preparation thereof Another object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which can be used for fast treatment in diseases like cold injuries, diabetes mellitus, Berger's disease, Raynaud's phenomenon, burns and other vasospastic, vasodestructive or ischemic conditions related to infective, autoimmune, neural, vascular, degenerative or inflammatory disorders. Yet another object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof that can be effective in extremes of atmospheric temperature, pressure and humidity. Further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which is easy to prepare. Still further an object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof that quickly increases the local blood circulation by virtue of vasodilation in the local microcirculation. Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which reduces the blood viscosity, reduce platelet aggregation, prevent intravascular clotting and helps in reducing oxidative injury on revascularization in the local microvasculature which, in turn, help in faster pain relief Still further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which utilises nitroglycerine or amylnitrate, both very potent and fast acting smooth-muscles relaxants, as the main active ingredient. Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which has no significant adverse-effects and can be used in cold and dry weather. Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which helps in relieving the painful ischemic symptoms, in preventing or reducing fiirther damage to the tissues, and in preventing or reducing oxidation injury. Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which causes early healing and increase work efficiency of the body part. SUMMARY OF THE INVENTION The present invention relates to a process for the preparation of topical formulation comprising the following steps: (a) preparing oil phase by mixing cetostearyl alcohol, cetomacrogel 1000 and liquid paraffin to white soft paraffin, followed by adding methyl paraben and propyl paraben melting and mixing these on water bath kept at a temperature between 45-65°C and adding dipyridamol; (b) preparing aqueous phase by dissolving polyethylene glycol in water 60-100 ml and heating preferably on a water bath preferably at less than 65 °C and adding active components Uke citrate, preferably citric acid niacin, preferably nicotinic acid and pentoxyphyllin to it before the heat treatment. (c) mixing the separately pre-heated oil phase prepared in step (a) and aqueous phase prepared in step (b) to less than 65°C cooling it to about 30-40°C and adding nitroglycerine or amylnitrite, glycerin and preferably ascorbic acid as powder under continuous stirring before adding a small quantity of compatible perfume. In one embodiment of the invention, the said cetostearyl alcohol is taken in an amount in the range of 1.2-13.3% by weight of the medicated ointment. In another embodiment of the invention, the liquid paraffin is taken in an amount in the range of 1.2-13.3% by weight of the topical formulation. In another embodiment of the invention, the white paraffin is taken in an amount in the range of 13.3-26.6% by weight of the said topical formulation. In another embodiment of the invention, the methyl and propyl paraben are taken in an amount of up to 0.4%) and up to 0.26% by weight respectively of the topical formulation. In another embodiment of the invention, the cetomacrogol 1000 is taken in an amount in the range of 0.6-4% by weight of the topical formulation. In another embodiment of the invention, the PEG 400 is taken in an amount in the range of 0.3-0.66 % by weight of the topical formulation. In another embodiment of the invention, the nitroglycerine or amylnitrite is taken in an amount in the range of 0.06- 1.33% by weight of the topical formulation. In another embodiment of the invention, the glycerin is taken in an amount in the range of 0.06%- 1.33%) by weight of the topical formulation. In another embodiment of the invention, the dipyridamole is taken in an amount of to 2.7% by weight of the topical formulation. In another embodiment of the invention, the nicotinic acid is taken in an amount of up to 4% by weight of the topical formulation. In another embodiment of the invention, the citric acid is taken in an amount in the range of 2-5% by weight of the topical formulation. In another embodiment of the invention, the pentoxyphyllin is taken in an amount of up to 6.6% by weight of the topical formulation. In another embodiment of the invention, the ascorbic acid is taken in an amount of up to 4% by weight of the topical formulation. In another embodiment of the invention, the balance in weight or volume of the topical formulation is provided by purified water The present invention also relates to a topical formulation comprising nitroglycerine or amylnitrite, cetostearyl alcohol, liquid paraffin, soft white paraffin, methyl and propyl paraben, cetomacrogol 1000, PEG 400, glycerin, dipyridamole, nicotinic acid, citric acid, pentoxyphyllin, ascorbic acid and purified water. In one embodiment of the invention, the formulation comprises nitroglycerine or amylnitrite 0.06-1.1% by weight, cetostearyl alcohal 1.2-13.3.1% by weight, Uquid paraffin 1.2-13.3% by weight, soft white paraffin 13.3-26.6% by weight, methyl and propyl paraben up to 0.4% and up to 0.26% respectively by weight, cetomacrogol 1000 0.6-4% by weigh, PEG 400 .03-0.66% by weight, glycerin 0.06-1.33% by weight, dipyridamole up to 2.7% by weight, nicitinic acid up to 4% by weight, citric acid 2-5% by weight, pentoxyphyllin up to 6.6% by weight, ascorbic acid-1.5%) by weight, and rest of the weight and volume by purified water. The formulation of the invention is a synergistic admixture whose properties are improved or unexpected and are not a mere aggregation of the properties of the individual ingredients. DETAILED DESCRIPTION OF THE INVENTION The present invention provides a process for the preparation of a topical formulation, which provides a potent and fast means to treat acute or severe forms of peripheral vascular diseases, particularly those involving micro-circulation of hands and feet. The topical formulation of the present invention is help fill in diseases hke cold injuries, diabetes mellitus, Berger's disease, Raynaud's phenomenon, burns and other vasospastic, vasodestructive or ischemic conditions related to infective, autoimmune, neural, vascular, degenerative or inflammatory disorders. The topical formulation, even after a single apphcation may increase the local blood supply, and may reduce inflammation, intravascular thrombosis and clotting, and viscosity of the blood, thus improving the blood flow in the lesion area. The formulation helps in relieving the painfiil ischemic symptoms, in preventing or reducing further damage to the tissues, and in preventing or reducing oxidation injury. It also causes early heating and increase work efficiency of the body part. The formulation has no significant adverse-effects and can be used in cold and dry weather. The main active ingredient of the invention is nitroglycerine or amyl nitrite, both very potent and fast-acting smooth-muscle relaxants, belonging to the same chemical group. Other preferred active ingredients of the topical formulation include citrate, particularly its acid, ascorbate, particularly its acid, and niacin, more preferably, nicotinic acid. Still other preferred ingredients are pentoxyphyllin and dipyridamole. Several inert base substances are also added. These are helpful in maintaining the active components in a state that ensures dermal permeability, sustained action and integrity of the ointment. The base materials include glycols, macrogels, paraffins, alcohols, preservatives, a hygroscopic agent and some perfome. The preferred solvent of the base materials is refined water, making the formulation an oil-in-water emulsion preparation. Copending Indian Patent Application No. filed on the same day as this application relates to a topical formulation prepared by the process of this invention. The invention of copending apphcation comprises essentially of a topical formulation comprising nitroglycerine or amylnitrite, cetostearyl alcohol, liquid paraffin, soft white paraffin, methyl and propyl paraben, cetomacrogol 1000, PEG 400, glycerin, dipyridamole, nicotinic acid, citric acid, pentoxyphyllin, ascorbic acid and purified water. In the topical formulation, he ingredients comprise nitroglycerine or amylnitrite 0.06-1.1% by weight, cetostearyl alcohal 1.2-13.3.1% by weight, liquid paraffin 1.2-13.3% by weight, soft white paraffin 13.3-26.6% by weight, methyl and propyl paraben up to 0.4% and up to 0.26% respectively by weight, cetomacrogol 1000 0.6-4% by weigh, PEG 400 .03-0.66% by weight, glycerin 0.06-1.33% by weight, dipyridamole up to 2.7% by weight, nicitinic acid up to 4% by weight, citric acid 2-5% by weight, pentoxyphyllin up to 6.6%) by weight, ascorbic acid-1.5%) by weight, and rest of the weight and volume by purified water. The multi-component topical formulation is prepared by making an oil-phase and aqueous phase separately and mixing the two in a determined fashion and adding the active ingredient(s) in a determinate manner. The oil phase and water phase are maintained at less than 65 °C. Aqueous phase is poured in oil phase with continuos stirring. The complex is cooled to 35°C or even lower after which glycerin and preferably ascorbic acid in powder form, and perfiime are added as the last step of the formulation, again with continuos stirring. However, the sequence of predetermined steps may be changed in manner not detrimental to the stability and efficacy of the medicated ointment product. The topical formulation of the present invention is prepared by the process comprising of the following steps: a) Preparation of oil phase The oil phase is prepared by mixing cetostearyl alcohol 1.2-13.3% wt wise, cetomacrogol 1000 0.6-4% wt wise and hquid paraffin 1.2-13.3% wt wise to white soft paraffin taken 13.3-36.6%) wt wise, followed by adding methyl paraben up to 0.4%i wt wise and propyl paraben up to 0.26%) wt wise, followed by melting and mixing these preferably on water bath kept at a temperature preferably between 45-65°C and adding dipyridamol up to 2.7% wt wise, b) Preparation of aqueous phase The aqueous phase is prepared by dissolving polyethylene glycol 400 0.3-0.66%) wt wise in water 60-100ml and heating preferably on a water bath preferably at less than 65°C. Other preferable active components like Citrate, preferably citric acid up to 6.6.%) wt wise., niacin, preferably nicotinic acid up to 4%) wt. wise and pentoxyphyllin up to 6.6%) wt wise are added to the aqueous phase before the heat treatment. C) Preparation of formulation Oil phase and the aqueous phase prepared as above area separately maintained preferably at a temperature less than 65°C. Next, aqueous phase is poured in oil phase with continuous stirring. The complex is cooled at 30-40°C following which nitroglycerine or amylnitrite taken 0.06-1.1%) wt wise, glycerin .06-133%) wt wise, and preferably ascorbic acid as powder up to 4% wt wise, are mixed with continuous stirring before adding a small quantity of compatible perfume. The invention will now be illustrated with a working example, which is intending to be an illustrate example, and is not intending to be taken restrictively to imply any limitation on the scope of the present invention. WORKING EXAMPLE 2.5 gram cetomacrogol 1000, 5 gram cetostearyl alcohol and 18 gram white soft paraffin are melted on water bath in the decreasing order of their melting points. 100 mg propyl paraben is added to 10 gram liquid paraffin and it was heated till it dissolved. Next, the above two mix are mixed together. Next, 200 mg PEG 400 is added in 75 gram water and heated on water bath. The oil phase and water phase are heated to 60°C. The aqueous phase is poured in oil phase with continuous stirring. After cooling it to 35°C, 100 mg nitroglycerine and 100 mg glycerin and perfiime are added. CLINICAL EVALUATION: Vasodilator action of the nitroglycerine/amylnitrite based multicomponent medicated ointment was tested using nuclear medicine technology in 8 healthy volunteers. Digital angiographic and / or blood-pool images of the hands and / or feet were taken after injecting a radiopharmaceutical on a Gamma or SPECT Camera. After applying the 2 gram of medicated ointment on the surface of one hand / feet and gently rubbing to spread it completely, the nuclear vascularity of the part has increased following the ointment application and at what interval. Typically, the imaging protocol was repeated before and after 1-3 days of application (two times a day) to evaluate the benefit qualitatively and quantitatively. All eight individuals showed more than 15% rise in circulation within 6 hrs of apphcation and more than 60% (60-110%)) increases in blood flow in the part of apphcation within 489 hrs of the topical application. All patients developed feeling of warmth over the part applied to. Few on these developed redness also. There were no side effects noted in any of the patients. Phase 1-2 field trial at high altitude in more than 15 patient volunteers has shown reduction in pain & numbness and increase in feeling of warmth & skin temperature by 0.5-2°C following topical application. No local or systemic side effects have been observed. It is to be understood that the formulation and process of the present invention is susceptible to changes, adaptations and modifications by those skilled in the art. Such variant embodiments that incorporate the features and concepts of the present invention are intended to be within the scope of the present invention, which is further set forth under the following claims: WE CLAIM: 1. A process for the preparation of topical formulation comprising the following steps: (a) preparing oil phase by mixing cetostearyl alcohol, cetomacrogel 1000 and liquid paraffin to white soft paraffin, followed by adding methyl paraben and propyl paraben melting and mixing these on water bath kept at a temperature between 45-65°C and adding dipyridamol; (b) preparing aqueous phase by dissolving polyethylene glycol in water 60-100 ml and heating preferably on a water bath preferably at less than 65°C and adding active components like citrate, preferably citric acid niacin, preferably nicotinic acid and pentoxyphyllin to it before the heat treatment. (c) mixing the separately pre-heated oil phase prepared in step (a) and aqueous phase prepared in step (b) to less than 65 °C cooling it to about 30-40°C and adding nitroglycerine or amylnitrite, glycerin and preferably ascorbic acid as powder under continuous stirring before adding a small quantity of compatible perfume. 2. A process as claimed in claim 1 wherein the oil phase and water phase are maintained at a temperature of less than 65°C. 3. A process as claimed in claim 1 wherein the aqueous phase is poured in oil phase with continuos stirring and the resultant complex cooled to 35°C or less after which glycerin, and preferably ascorbic acid in powder form, and perfiime, are added with continuos stirring. 4. A process as claimed in claim 1 wherein the oil phase is prepared by mixing cetostearyl alcohol 1.2-13.3% wt wise, cetomacrogol 1000 0.6-4% wt wise and hquid paraffin 1.2-13.3% wt wise to white soft paraffin taken 13.3-36.6% wt wise, followed by adding methyl paraben up to 0.4% wt wise and propyl paraben up to 0.26%) wt wise, followed by melting and mixing these preferably on water bath kept at a temperature preferably between 45-65°C and adding dipyridamol up to 2.7%) wt wise. 5. A process as claimed in claim 1 wherein the aqueous phase is prepared by dissolving polyethylene glycol 400 0.3-0.66% wt wise in water 60-l00ml and heating preferably on a water bath preferably at less than 65°C. 6. A process as claimed in claim 1 wherein the active components like Citrate, preferably citric acid up to 6.6.% wt wise., niacin, preferably nicotinic acid up to 4% wt. wise and pentoxyphyllin up to 6.6% wt wise are added to the aqueous phase before heat treatment. 7. A process as claimed in claim 1 wherein the formulation is prepared by maintaining the oil phase and the aqueous phase separately preferably at a temperature less than 65 °C, the aqueous phase then being poured into the oil phase under continuous stirring and the resultant complex being cooled to a temperature in the range of 30-40°C following which nitroglycerine or amylnitrite taken 0.06-1.1% wt wise, glycerin .06-133% wt wise, and preferably ascorbic acid as powder up to 4% wt wise, are mixed with continuous stirring before adding a small quantity of compatible perfume. 8. A process as claimed in claim 1 wherein the said cetostearyl alcohol is taken in the quantity of 1.2-13.3% by weight of the medicated ointment. 9. A process as claimed in claim 1 wherein the said liquid paraffin is taken in the quantity of 1.2-13.3% by weight of the said topical formulation. 10. A process as claimed in claim 1 wherein the said white paraffin is taken in the quantity of 13.3-26.6% by weight of the said topical formulation. 11. A process as claimed in claim 1 wherein the said methyl and propyl paraben are taken in the quantity up to 0.4% and up to 0.26%) by weight respectively of the said topical formulation. 12. A process as claimed in claim 1 wherein the said cetomacrogol 1000 is taken in the quantity of 0.6-4%) by weight of the said topical formulation. 13. A process as claimed in claim 1 wherein the said PEG 400 is taken in the quantity of 0.3-0.66 %) by weight of the said topical formulation. 14. A process as claimed in claim 1 wherein the said nitroglycerine or amylnitrite is taken in the quantity of 0.06- 1.33%) by weight of the said topical formulation. 15. A process as claimed in claim 1 wherein the said glycerin is taken in the quantity of 0.06%-1.33%) by weight of the said topical formulation. 16. A process as claimed in claim 1 wherein the said dipyridamole is taken in the quantity up to 2.7%) by weight of the said topical formulation. 17. A process as claimed in claim 1 wherein the said nicotinic acid is taken in the quantity up to 4% by weight of the said topical formulation. 18. A process as claimed in claim 1 wherein the said citric acid is taken in the quantity of 2-5% by weight of the said topical formulation. 19. A process as claimed in claim 1 wherein the said pentoxyphyllin is taken in the quantity of 6.6%) by weight of the said topical formulation. 20. A process as claimed in claim 1 wherein the said ascorbic acid is taken in the quantity of 4% by weight of the said topical formulation. 21. A process as claimed in claim 1 wherein the rest of the weight and volume of said topical formulation is provided by purified water 22. A process for preparing a topical formulation substantially as described and with reference to the foregoing examples.

Full Text

PROCESS FOR THE PREPARATION OF A NITRO-GROUP BASED TOPICAL FORMULATION FOR TREATMENT OF ACUTE PERIPHERAL MICROVASCULAR DISEASES
FIELD OF THE INVENTION
The present invention relates to a process for the preparation of a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. The present invention also relates to a nitro-group based topical formulation for treatment of cold injuries and other peripheral vascular diseases. More particularly, the present invention relates to a nitro group based topical formulation for the treatment of cold injuries and other peripheral vascular diseases such as those afflicting microvasculature or microcirculation. BACKGROUND OF THE INVENTION
Vasospastic/vasodestructive or ischemic disorders of limbs are very common. These are found even in apparently normal individuals, and generally occur due to transient or permanent reduction of blood supply and oxygen at tissue level. Such disorders occur more frequently and severely in cold injuries, diabetes mellitus, Berger's disease, Raynaud's phenomenon, burns, arteritis of all types and other such diseases coming under the broad category of peripheral vascular diseases, particularly involving microcirculation or microvasculature. Hypovolemia, atherosclerosis, anemia and old age are further risk factors. Thrombosis in microvasculature of hands or feet causes additional complications, leading to further reduction in blood and oxygen supply and permanent damage of the tissues.
Such disorders can lead to tissue death, gangrene and loss of limb or function. Such conditions are further aggravated in cold weather, particularly at high altitude and in old individuals and those who consume tobacco products or are exposed to second hand effects of tobacco products. Another related problem or complication is that local ischemia leads to development of infection and inflammation that further causes more local ischemia resulting in a vicious cycle. These peripheral vascular diseases could be of different pathological origins, namely autoimmune, infective, environmental, toxic, autonomic, vascular, idiopathic and of other and unknown origins.
The disorders as described above lead to acute pain, cyanosis and loss of work efficiency (particularly of arms and legs, hands and fingers, feet and toes or facial parts) leading to edema and ulcer formation, leading to muscle and bone death, deep-seated infection, amputation and ultimately loss of the affected body organ. Even when the local blood and oxygen supply is partially restored by conventional treatment, the tissue injury aggravates due to oxidative mechanisms. Barring very mild injuries, the damage at

microvascular or micro-circulation level is permanent and may be progressive. Most of the predisposing or precipitating factors are either accidental or unpreventable.
It is known in the art to treat peripheral vascular diseases, particularly cold injuries in a non-pharmacological manner such as by slow re-warming of the affected part in water, called thawing. A practical limitation with of the above method is that this cannot be used as a preventive routinely but can only be used as to manage the condition. Another drawback is that it is neither practical nor possible to use thawing as the main course of treatment in field conditions. Another limitation of the slow re-warming is that the method is useful only for cold injuries and not other forms of peripheral vasospastic/ ischemic diseases. Even in cold injuries, thawing is useful for only mild cold injuries and is inadequate in major cold injuries such as frostbite. Thawing is also a very slow method of treatment and does not provide speedy relief
Various pharmacological methods are also known in the art for treatment of ischemic or vasospastic/vasodestructive disorders. Such pharmacological methods include systemic methods such as oral, intramuscular or intravenous administration of vasodilator drugs.
However, such pharmacological methods suffer from several limitations. A major limitation of the above method is that there is disagreement about its benefit in treating local micro-vascular conditions (mostly of arms and legs or hands and feet). This is primarily due to the inability of the systemic route to deliver sufficient quantity of medication to the peripheral ischemic lesion because of large volume of distribution. One of the most potent and fast-acting vasodilators, nitroglycerine or amylnitrate, if given by systemic route for local action on the aforesaid lesions, would result in very high pharmacological dose in the blood and is therefore impractical.
Another significant disadvantage of pharmacological methods are that there may not be any benefit at all if blood supply to the hand/foot is already blocked significantly due to thrombosis. Additionally, the dosage required for systemic route is high and can cause serious side effects such as low blood pressure and significant palpitation and heart problems. This limits the duration of treatment to a few days, thereby limiting the benefits of such treatment, if any. Thus, this route cannot deliver potent and fast-acting vasodilators such as nitroglycerine, even though these would be quite effective for lesion treatment. This method also results in high treatment cost due to the high dosages required for managing the disease.
Another method known in the art for management of the above diseases comprise systemic administration of anti-coagulants and/or thrombolytic agents. However, similar to systemic administration of vasodilators, the above method of management has the drawback
that it requires a very high dosage thereby increasing side effects (bleeding from unrelated sites) and cost. The use of anti-coagulants and thrombolytic drugs are also considered dangerous except in hospital conditions. Another drawback is that these agents cannot be used in chronic or recurring conditions. Their use at present is therefore limited to very severe cases considering the risks involved.
Another method known in the art for management of the above diseases is systemic administration of antiplatelet drugs Uke dipyridamole or aspirin. Similar to systemic administration of vasodilators, the above method of management has the drawback that it requires a very high dosage thereby increasing side effects and cost. Another drawback of the above method is that the pharmacological effect of such drugs is limited if the local blood circulation is compromised due to thrombosis.
It is also known in the art to manage the above diseases using oral pentoxyphyllin or other Xanthine-based drugs or Dextran infusion that are known to reduce blood viscosity, thus enabling more flow of blood to the lesions. However, the above method, like systemic vasodilators and antiplatelet drugs, has the drawback that a very high blood concentration is required for optimal concentration in the peripheral lesions, thus increasing side effects and cost. Also, it may have limited pharmacological effect if local blood circulation is compromised due to thrombosis.
Topical ointments containing analgesics and antibiotics are available but have the drawback that these are useful in controlling only the infection and do not address the main problem of microvascular insufficiency. Topical ointments containing nicotinic acid as a vasodilator (for example, 'ThrombophobeTM) are available in the market for treating microvascular insufficiency. Their main drawback is that though useful, these are usually not found sufficiently potent to cure the conditions by themselves and are only used for supportive treatment.
As such, there is need of a topical ointment which could penetrate effectively through the skin to deliver fast-acting and potent drugs that act on legal microvasculature and increase the local blood flow through vasodilation and other effects, OBJECTS OF THE EVVENTION
The main object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process for preparation thereof
Another object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which can be used for fast treatment in diseases like cold injuries, diabetes mellitus, Berger's disease, Raynaud's
phenomenon, burns and other vasospastic, vasodestructive or ischemic conditions related to infective, autoimmune, neural, vascular, degenerative or inflammatory disorders.
Yet another object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof that can be effective in extremes of atmospheric temperature, pressure and humidity.
Further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which is easy to prepare.
Still further an object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof that quickly increases the local blood circulation by virtue of vasodilation in the local microcirculation.
Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which reduces the blood viscosity, reduce platelet aggregation, prevent intravascular clotting and helps in reducing oxidative injury on revascularization in the local microvasculature which, in turn, help in faster pain relief
Still further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which utilises nitroglycerine or amylnitrate, both very potent and fast acting smooth-muscles relaxants, as the main active ingredient.
Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which has no significant adverse-effects and can be used in cold and dry weather.
Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which helps in relieving the painful ischemic symptoms, in preventing or reducing fiirther damage to the tissues, and in preventing or reducing oxidation injury.
Yet further object of the invention is to provide a topical formulation for fast treatment of peripheral vascular diseases and a process of preparation thereof which causes early healing and increase work efficiency of the body part. SUMMARY OF THE INVENTION
The present invention relates to a process for the preparation of topical formulation comprising the following steps:
(a) preparing oil phase by mixing cetostearyl alcohol, cetomacrogel 1000 and liquid
paraffin to white soft paraffin, followed by adding methyl paraben and propyl paraben
melting and mixing these on water bath kept at a temperature between 45-65°C and adding dipyridamol;
(b) preparing aqueous phase by dissolving polyethylene glycol in water 60-100 ml and heating preferably on a water bath preferably at less than 65 °C and adding active components Uke citrate, preferably citric acid niacin, preferably nicotinic acid and pentoxyphyllin to it before the heat treatment.
(c) mixing the separately pre-heated oil phase prepared in step (a) and aqueous phase prepared in step (b) to less than 65°C cooling it to about 30-40°C and adding nitroglycerine or amylnitrite, glycerin and preferably ascorbic acid as powder under continuous stirring before adding a small quantity of compatible perfume.
In one embodiment of the invention, the said cetostearyl alcohol is taken in an amount in the range of 1.2-13.3% by weight of the medicated ointment.
In another embodiment of the invention, the liquid paraffin is taken in an amount in the range of 1.2-13.3% by weight of the topical formulation.
In another embodiment of the invention, the white paraffin is taken in an amount in the range of 13.3-26.6% by weight of the said topical formulation.
In another embodiment of the invention, the methyl and propyl paraben are taken in an amount of up to 0.4%) and up to 0.26% by weight respectively of the topical formulation.
In another embodiment of the invention, the cetomacrogol 1000 is taken in an amount in the range of 0.6-4% by weight of the topical formulation.
In another embodiment of the invention, the PEG 400 is taken in an amount in the range of 0.3-0.66 % by weight of the topical formulation.
In another embodiment of the invention, the nitroglycerine or amylnitrite is taken in an amount in the range of 0.06- 1.33% by weight of the topical formulation.
In another embodiment of the invention, the glycerin is taken in an amount in the range of 0.06%- 1.33%) by weight of the topical formulation.
In another embodiment of the invention, the dipyridamole is taken in an amount of to 2.7% by weight of the topical formulation.
In another embodiment of the invention, the nicotinic acid is taken in an amount of up to 4% by weight of the topical formulation.
In another embodiment of the invention, the citric acid is taken in an amount in the range of 2-5% by weight of the topical formulation.
In another embodiment of the invention, the pentoxyphyllin is taken in an amount of up to 6.6% by weight of the topical formulation.
In another embodiment of the invention, the ascorbic acid is taken in an amount of up to 4% by weight of the topical formulation.
In another embodiment of the invention, the balance in weight or volume of the topical formulation is provided by purified water
The present invention also relates to a topical formulation comprising nitroglycerine or amylnitrite, cetostearyl alcohol, liquid paraffin, soft white paraffin, methyl and propyl paraben, cetomacrogol 1000, PEG 400, glycerin, dipyridamole, nicotinic acid, citric acid, pentoxyphyllin, ascorbic acid and purified water.
In one embodiment of the invention, the formulation comprises nitroglycerine or amylnitrite 0.06-1.1% by weight, cetostearyl alcohal 1.2-13.3.1% by weight, Uquid paraffin 1.2-13.3% by weight, soft white paraffin 13.3-26.6% by weight, methyl and propyl paraben up to 0.4% and up to 0.26% respectively by weight, cetomacrogol 1000 0.6-4% by weigh, PEG 400 .03-0.66% by weight, glycerin 0.06-1.33% by weight, dipyridamole up to 2.7% by weight, nicitinic acid up to 4% by weight, citric acid 2-5% by weight, pentoxyphyllin up to 6.6% by weight, ascorbic acid-1.5%) by weight, and rest of the weight and volume by purified water.
The formulation of the invention is a synergistic admixture whose properties are improved or unexpected and are not a mere aggregation of the properties of the individual ingredients. DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a process for the preparation of a topical formulation, which provides a potent and fast means to treat acute or severe forms of peripheral vascular diseases, particularly those involving micro-circulation of hands and feet. The topical formulation of the present invention is help fill in diseases hke cold injuries, diabetes mellitus, Berger's disease, Raynaud's phenomenon, burns and other vasospastic, vasodestructive or ischemic conditions related to infective, autoimmune, neural, vascular, degenerative or inflammatory disorders. The topical formulation, even after a single apphcation may increase the local blood supply, and may reduce inflammation, intravascular thrombosis and clotting, and viscosity of the blood, thus improving the blood flow in the lesion area. The formulation helps in relieving the painfiil ischemic symptoms, in preventing or reducing further damage to the tissues, and in preventing or reducing oxidation injury. It also causes early heating and increase work efficiency of the body part. The formulation has no significant adverse-effects and can be used in cold and dry weather.
The main active ingredient of the invention is nitroglycerine or amyl nitrite, both very potent and fast-acting smooth-muscle relaxants, belonging to the same chemical group. Other preferred active ingredients of the topical formulation include citrate, particularly its acid, ascorbate, particularly its acid, and niacin, more preferably, nicotinic acid. Still other preferred ingredients are pentoxyphyllin and dipyridamole. Several inert base substances are also added. These are helpful in maintaining the active components in a state that ensures dermal permeability, sustained action and integrity of the ointment. The base materials include glycols, macrogels, paraffins, alcohols, preservatives, a hygroscopic agent and some perfome. The preferred solvent of the base materials is refined water, making the formulation an oil-in-water emulsion preparation.
Copending Indian Patent Application No. filed on the same day as
this application relates to a topical formulation prepared by the process of this invention. The invention of copending apphcation comprises essentially of a topical formulation comprising nitroglycerine or amylnitrite, cetostearyl alcohol, liquid paraffin, soft white paraffin, methyl and propyl paraben, cetomacrogol 1000, PEG 400, glycerin, dipyridamole, nicotinic acid, citric acid, pentoxyphyllin, ascorbic acid and purified water. In the topical formulation, he ingredients comprise nitroglycerine or amylnitrite 0.06-1.1% by weight, cetostearyl alcohal 1.2-13.3.1% by weight, liquid paraffin 1.2-13.3% by weight, soft white paraffin 13.3-26.6% by weight, methyl and propyl paraben up to 0.4% and up to 0.26% respectively by weight, cetomacrogol 1000 0.6-4% by weigh, PEG 400 .03-0.66% by weight, glycerin 0.06-1.33% by weight, dipyridamole up to 2.7% by weight, nicitinic acid up to 4% by weight, citric acid 2-5% by weight, pentoxyphyllin up to 6.6%) by weight, ascorbic acid-1.5%) by weight, and rest of the weight and volume by purified water.
The multi-component topical formulation is prepared by making an oil-phase and aqueous phase separately and mixing the two in a determined fashion and adding the active ingredient(s) in a determinate manner.
The oil phase and water phase are maintained at less than 65 °C. Aqueous phase is poured in oil phase with continuos stirring. The complex is cooled to 35°C or even lower after which glycerin and preferably ascorbic acid in powder form, and perfiime are added as the last step of the formulation, again with continuos stirring. However, the sequence of predetermined steps may be changed in manner not detrimental to the stability and efficacy of the medicated ointment product.
The topical formulation of the present invention is prepared by the process comprising of the following steps:
a) Preparation of oil phase
The oil phase is prepared by mixing cetostearyl alcohol 1.2-13.3% wt wise, cetomacrogol 1000 0.6-4% wt wise and hquid paraffin 1.2-13.3% wt wise to white soft paraffin taken 13.3-36.6%) wt wise, followed by adding methyl paraben up to 0.4%i wt wise and propyl paraben up to 0.26%) wt wise, followed by melting and mixing these preferably on water bath kept at a temperature preferably between 45-65°C and adding dipyridamol up to 2.7% wt wise, b) Preparation of aqueous phase
The aqueous phase is prepared by dissolving polyethylene glycol 400 0.3-0.66%) wt wise in water 60-100ml and heating preferably on a water bath preferably at less than 65°C. Other preferable active components like Citrate, preferably citric acid up to 6.6.%) wt wise., niacin, preferably nicotinic acid up to 4%) wt. wise and pentoxyphyllin up to 6.6%) wt wise are added to the aqueous phase before the heat treatment. C) Preparation of formulation
Oil phase and the aqueous phase prepared as above area separately maintained preferably at a temperature less than 65°C. Next, aqueous phase is poured in oil phase with continuous stirring. The complex is cooled at 30-40°C following which nitroglycerine or amylnitrite taken 0.06-1.1%) wt wise, glycerin .06-133%) wt wise, and preferably ascorbic acid as powder up to 4% wt wise, are mixed with continuous stirring before adding a small quantity of compatible perfume.
The invention will now be illustrated with a working example, which is intending to be an illustrate example, and is not intending to be taken restrictively to imply any limitation on the scope of the present invention. WORKING EXAMPLE
2.5 gram cetomacrogol 1000, 5 gram cetostearyl alcohol and 18 gram white soft paraffin are melted on water bath in the decreasing order of their melting points. 100 mg propyl paraben is added to 10 gram liquid paraffin and it was heated till it dissolved. Next, the above two mix are mixed together. Next, 200 mg PEG 400 is added in 75 gram water and heated on water bath. The oil phase and water phase are heated to 60°C. The aqueous phase is poured in oil phase with continuous stirring. After cooling it to 35°C, 100 mg nitroglycerine and 100 mg glycerin and perfiime are added. CLINICAL EVALUATION:
Vasodilator action of the nitroglycerine/amylnitrite based multicomponent medicated ointment was tested using nuclear medicine technology in 8 healthy volunteers. Digital
angiographic and / or blood-pool images of the hands and / or feet were taken after injecting a radiopharmaceutical on a Gamma or SPECT Camera. After applying the 2 gram of medicated ointment on the surface of one hand / feet and gently rubbing to spread it completely, the nuclear vascularity of the part has increased following the ointment application and at what interval. Typically, the imaging protocol was repeated before and after 1-3 days of application (two times a day) to evaluate the benefit qualitatively and quantitatively. All eight individuals showed more than 15% rise in circulation within 6 hrs of apphcation and more than 60% (60-110%)) increases in blood flow in the part of apphcation within 489 hrs of the topical application. All patients developed feeling of warmth over the part applied to. Few on these developed redness also. There were no side effects noted in any of the patients.
Phase 1-2 field trial at high altitude in more than 15 patient volunteers has shown reduction in pain & numbness and increase in feeling of warmth & skin temperature by 0.5-2°C following topical application. No local or systemic side effects have been observed.
It is to be understood that the formulation and process of the present invention is susceptible to changes, adaptations and modifications by those skilled in the art. Such variant embodiments that incorporate the features and concepts of the present invention are intended to be within the scope of the present invention, which is further set forth under the following claims:

WE CLAIM:
1. A process for the preparation of topical formulation comprising the following steps:
(a) preparing oil phase by mixing cetostearyl alcohol, cetomacrogel 1000 and liquid paraffin to white soft paraffin, followed by adding methyl paraben and propyl paraben melting and mixing these on water bath kept at a temperature between 45-65°C and adding dipyridamol;
(b) preparing aqueous phase by dissolving polyethylene glycol in water 60-100 ml and heating preferably on a water bath preferably at less than 65°C and adding active components like citrate, preferably citric acid niacin, preferably nicotinic acid and pentoxyphyllin to it before the heat treatment.
(c) mixing the separately pre-heated oil phase prepared in step (a) and aqueous phase prepared in step (b) to less than 65 °C cooling it to about 30-40°C and adding nitroglycerine or amylnitrite, glycerin and preferably ascorbic acid as powder under continuous stirring before adding a small quantity of compatible perfume.

2. A process as claimed in claim 1 wherein the oil phase and water phase are maintained at a temperature of less than 65°C.
3. A process as claimed in claim 1 wherein the aqueous phase is poured in oil phase with continuos stirring and the resultant complex cooled to 35°C or less after which glycerin, and preferably ascorbic acid in powder form, and perfiime, are added with continuos stirring.
4. A process as claimed in claim 1 wherein the oil phase is prepared by mixing cetostearyl alcohol 1.2-13.3% wt wise, cetomacrogol 1000 0.6-4% wt wise and hquid paraffin 1.2-13.3% wt wise to white soft paraffin taken 13.3-36.6% wt wise, followed by adding methyl paraben up to 0.4% wt wise and propyl paraben up to 0.26%) wt wise, followed by melting and mixing these preferably on water bath kept at a temperature preferably between 45-65°C and adding dipyridamol up to 2.7%) wt wise.
5. A process as claimed in claim 1 wherein the aqueous phase is prepared by dissolving polyethylene glycol 400 0.3-0.66% wt wise in water 60-l00ml and heating preferably on a water bath preferably at less than 65°C.
6. A process as claimed in claim 1 wherein the active components like Citrate, preferably citric acid up to 6.6.% wt wise., niacin, preferably nicotinic acid up to 4% wt. wise and pentoxyphyllin up to 6.6% wt wise are added to the aqueous phase before heat treatment.
7. A process as claimed in claim 1 wherein the formulation is prepared by maintaining the oil phase and the aqueous phase separately preferably at a temperature less than 65 °C, the

aqueous phase then being poured into the oil phase under continuous stirring and the resultant complex being cooled to a temperature in the range of 30-40°C following which nitroglycerine or amylnitrite taken 0.06-1.1% wt wise, glycerin .06-133% wt wise, and preferably ascorbic acid as powder up to 4% wt wise, are mixed with continuous stirring before adding a small quantity of compatible perfume.
8. A process as claimed in claim 1 wherein the said cetostearyl alcohol is taken in the quantity of 1.2-13.3% by weight of the medicated ointment.
9. A process as claimed in claim 1 wherein the said liquid paraffin is taken in the quantity of 1.2-13.3% by weight of the said topical formulation.
10. A process as claimed in claim 1 wherein the said white paraffin is taken in the quantity of 13.3-26.6% by weight of the said topical formulation.
11. A process as claimed in claim 1 wherein the said methyl and propyl paraben are taken in the quantity up to 0.4% and up to 0.26%) by weight respectively of the said topical formulation.
12. A process as claimed in claim 1 wherein the said cetomacrogol 1000 is taken in the quantity of 0.6-4%) by weight of the said topical formulation.
13. A process as claimed in claim 1 wherein the said PEG 400 is taken in the quantity of 0.3-0.66 %) by weight of the said topical formulation.
14. A process as claimed in claim 1 wherein the said nitroglycerine or amylnitrite is taken in the quantity of 0.06- 1.33%) by weight of the said topical formulation.
15. A process as claimed in claim 1 wherein the said glycerin is taken in the quantity of 0.06%-1.33%) by weight of the said topical formulation.
16. A process as claimed in claim 1 wherein the said dipyridamole is taken in the quantity up to 2.7%) by weight of the said topical formulation.
17. A process as claimed in claim 1 wherein the said nicotinic acid is taken in the quantity up to 4% by weight of the said topical formulation.
18. A process as claimed in claim 1 wherein the said citric acid is taken in the quantity of 2-5% by weight of the said topical formulation.
19. A process as claimed in claim 1 wherein the said pentoxyphyllin is taken in the quantity of 6.6%) by weight of the said topical formulation.
20. A process as claimed in claim 1 wherein the said ascorbic acid is taken in the quantity of 4% by weight of the said topical formulation.
21. A process as claimed in claim 1 wherein the rest of the weight and volume of said topical formulation is provided by purified water

22. A process for preparing a topical formulation substantially as described and with reference to the foregoing examples.

Documents:

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Patent Number

270048

Indian Patent Application Number

2191/DEL/2004

PG Journal Number

48/2015

Publication Date

27-Nov-2015

Grant Date

26-Nov-2015

Date of Filing

03-Nov-2004

Name of Patentee

THE DIRECTOR GENERAL-DEFENCE RESEARCH & DEVELOPMENT ORGANISATION.

Applicant Address

MINISTRY OF DEFENCE, GOVERNMENT OF INDIA, OF WEST BLOCK-8, WING-1, SECTOR-1, RK PURAM, NEW DELHI - 110 066, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	ASEEM BHATNAGAR	INSTITUTE OF NUCLEAR MEDICINE& ALLIED SCIENCES, DRDO, MINISTRY OF DEFENCE., DELHI, INDIA.
2	NIDHI BHARDWAJ	INSTITUTE OF NUCLEAR MEDICINE & ALLIED SCIENCES, DRDO, MINISTRY OF DEFENCE, DELHI, INDIA.
3	AJAY KUMAR SINGH	INSTITUTE OF NUCLEAR MEDICINE & ALLIED SCIENCES, DRDO, MINISTRY OF DEFENCE, DELHI, INDIA.
4	ROOP KUMAR KHAR	DEPTT OF PHARMACEUTICALS, JAMIA HAMDARD UNIVERSITY, NEW DELHI, INDIA.
5	WILLIAM SELWAMURTHY	DRDO HEADQUARTERS, SENA BHAWAN, NEW DELHI - 110 011, INDIA.

PCT International Classification Number

A61K 3/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA