Title of Invention	A PROCESS OF PREPARATION OF ANTICANDIDIASIS AGENT FROM DRY MATURE SEEDS OF MYRISTICA FRAGRANS
Abstract	The present invention is about control of Candidiasis by ethanol extract of mature dry seeds of Myristica fragrans. Candidiasis is a disease caused by a group of Candida species, specially Candida albicans which is a normal saprophytic inhabitant of certain mucocutaneous surfaces of the body. Its overgrowth under certain conditions causes the disease. This is treated by nystatin, topical azole and fluconazole. But these drugs never cure the disease leaving high incidence of reoccurrence. In the present invention, the drug when applied to the affected areas gradually cured the disease without leaving any chance lor reoccurrence. Such cure of the disease has become possible for the first time as till to date no other drug has been able to cure this disease though the traditional antifungal drugs reduce the severity of the disease.

Title of Invention

A PROCESS OF PREPARATION OF ANTICANDIDIASIS AGENT FROM DRY MATURE SEEDS OF MYRISTICA FRAGRANS

Abstract

The present invention is about control of Candidiasis by ethanol extract of mature dry seeds of Myristica fragrans. Candidiasis is a disease caused by a group of Candida species, specially Candida albicans which is a normal saprophytic inhabitant of certain mucocutaneous surfaces of the body. Its overgrowth under certain conditions causes the disease. This is treated by nystatin, topical azole and fluconazole. But these drugs never cure the disease leaving high incidence of reoccurrence. In the present invention, the drug when applied to the affected areas gradually cured the disease without leaving any chance lor reoccurrence. Such cure of the disease has become possible for the first time as till to date no other drug has been able to cure this disease though the traditional antifungal drugs reduce the severity of the disease.

Full Text	COMPLETE SPECIFICATION Title of the invention: Control of Camlidiasis by ethanol extract of Myristica fragrans Objectives of invention The objective of this invention is to provide a drug, which can control Candidiasis in human beings and also its recurrence without any side effects. Myristica fragrans (Nutmeg) The principal aromatic constituent of the volatile oil of nutmeg, the dry ripe seeds of M. fragrans is myristicin or methoxysafrole. This yields 7-16% volatile oil which contains approximately 4-8% myristicin, the nutmeg contains approximately 1.3% myristicin. In addition to it, the seed contains many other compounds also (Leung, 1980). The chemical name of myristicin is 1 -allyl-5-methoxy-3, 4 methylene-dioxybenzene; CAS No. 607-91-0; empirical formula: C11H12O3; molecular weight: 192-22 (Merck Index, 1989). CANDIDIASIS Candidiasis is a fungal infection caused by related group of yeasts which are normal saprophytic inhabitants of mucocutancous body surfaces. They overgrow and invade tissues when permitted by alterations in the host. The causative organism is usually Candida albicans. Its manifestations may be localized to the skin or it may even be systemic and life threatening. Candidiasis occurs under the circumstances when a person 1. Suffers from diseases that either involve the immune system primarily or require therapies that adversely affect immune competence. 2. Treated intensively with antibacterial agents, glucocorticoids or use of oral contraceptives. 3. Has been inoculated intravenously, thus bypassing normal defences. Candidal infections have affinity for sites that are commonly wet and macerated for example corners of the mouth, oral cavity, around nails, intertriginous areas, penis, scrotum, inner aspects of thighs etc. In infants, cutaneous infections occur primarily as diaper rashes. This disease is frequently accompanied by a marked inflammatory response. Almost all cases of oropharyngeal, gastrointestinal, vaginal, systemic and cutaneous Candidiasis are caused by C. albicans. TREATMENT It involves removal of predisposing factors such as systemic antibiotic, glucocorticoids therapy, chronic wetness and the use of appropriate topical azoles and systemic fluconazole, if required in severe cases. The dose of fluconazole depends on the site and severity of the disease. Description of invention The dry mature seeds of Myrislica fragrans were powdered in a pastle and mortar to make powder. One kilogram of powder was immersed in 1.5 1 of ethanol in a conical flask and kept at room temperature (25 ±2° C) for a period of 10 days. The flask was shaken manually several times daily for 10 days. After this period the material was filtered through cotton and the filtrate was collected in glass beakers. The filtrate was left in the beaker for one week at the above temperature for evaporation of ethanol. At the end of one week, the pH of the filtrate was 5.5 and the optical density was 760 nm.. This ethanol extract (drug) was applied locally to those patients who were suffering from Candidiasis in intertriginous area, around nails and were resistant to topical and systemic antifugal therapy. With the traditional antifungal therapy, the disease used to subside for some time and then again recur with severe itching, erythema and maceration of the affected area which used to be hyperkeratitic with treatment but never disappear. The application of this drug in such cases stopped itching within 2 days. Within a week the erythema completely disappeared with brownish discoloration of the area and the thickness and diameter of the hyperkeratotic plaque started reducing. After approximately two weeks the plaque reduced by 40%. Within four weeks of application, the plaque was hardly visible. Within next few days, there was complete healing of the affected area. This effect of the present drug seems to be miracle for the treatment of Candidiasis because with the traditional local and systemic antifugal therapy the plaque do not respond though the itching and erythema of the disease respond. This persistent hyperkeratotic plaque becomes responsible for recurrence of the disease. Our drug heals the affected skin to such an extent that the plaques disappear completely and thus do not leave any chance of recurrence of the disease on that site. This effective action of this drug provides us with a comprehensive alternative to the other established modes of treatment, by curing the disease. Present national and international knowledge Many scientists have reported different properties of the extracts of dry seeds of M.fragrans. Sell and Carlini (1976) reported anaesthetic effect of methyl-euganol and other euganol derivatives of the volatile oil fraction of M. fragrans. In Nigeria and Indonesia, the essential oil of nutmeg is used in rheumatism (Oliver-Berver, 1986, Gils and Cox, 1994), probably because the oil decreases the level of prostaglandins which are major mediators of the inflammatory action (Iwo, 1983). This is also a known chemopreventor of chemically induced carcinogenesis (Singh and Rao, 1993). Us cytotoxic effect on cancer cells in vitro has been reported by Park et al (1998). Its antibacterial activity against Staphylococcnx aurens has been reported by Parez and Anesini (1994). Sharma et al. (1995) and Ram el al (1996) have reported prevention of hypercholesterolemia and atherosclerosis in rabbits after supplementation with seed extract. Olajide et al. (1999) described biological effects of chloroform extract of M. fragrans in male Swiss albino mice and male wistar rats as analgesic, anti-inflammatory and antithrombotic, whereas antidiarrheal activity of crude nutmeg solution as well as its petroleum ether extract has been reported by Grover et al. (2002). Myristicin which is the majopr component of essential oil of nutmeg is reported to posses extraordinarily potent hepatoprotective activity in mice against lipopolysaccharide (LPS) plus d-galactosamine (D-GalN) induced hepatotoxicity (Morita ci al. 2003). Though it has several other properties its toxicological effects have not been proved as yet (Hall Strom and Thuvander, 1997). There is also till today no report about the effect of ethanol extract of M.fragrans against Candidiasis of plaque type. Hence, this forms the first report in this direction establishing completely new invention which will prove to be the most important and effective treatment of Candidiasis without any side effects. Present state of development The ethanol extract of dry mature seeds of M.fragrans has been used in several patients suffering from Candidiasis with surprisingly complete healing effect. Utility of the invention The high rate of occurrence and recurrence of Cadidiasis is well known in spite of its treatment by traditional local and systemic antifugal drugs. The present drug starts acting by reducing the pruritis in Candidiasis. This is followed by reduction of erythema due to its anti-inflammatory action. Gradually this starts healing the macerated hyperkeratotic plaque and within a period of one month it completely heals leaving no chance for reoccurrence of the disease which is not possible by any other drug. It may also be possible because of its anti Candida albicans (antifungal) action. Since this drug is derived from an edible spice, which is commonly used in Indian kitchen, it may also be formulated for oral administration for the control of this disease. Till date its toxic effects have not been reported unless given in very high doses. Novelty and nonobviousness The effect of ethanol extract of M.fragmns is a completely new invention. It was not known earlier to this invention as is obvious from the review of literature till date regarding this property. This drug is highly effective against Candidiasis. No other drug is known to treat this disease completely once it occurs. Hence, this drug seems to be much better and complete option for the treatment and cure of this disease. DESCRIPTION OF THE INVENTION The dry mature seeds of Myristica fragrans were powdered in a pastle and mortar. One kilogram of this powder was immersed in 1.5 1 of ethanol in a conical flask and kept at room temperature (25 ±2° C) for a period of 10 days. During this period the flask was repeatedly shaken and then the material was filtered through cotton and the filtrate was collected in glass beakers. It was left open at room temperature for one week. The pH of the filtrate was measured which was 5.5 and the optical density was 760 nm. Thus obtained ethanol extract (drug) was tested for the treatment of Candidiasis also in those patients who had local and systemic antifungal therapy. It was gently applied to the affected areas thrice a day in both the groups. First application of the drug was soothing. Within two days the itching stopped completely. Within a week the erythema completely disappeared and the hyperkeratotic plaque started healing which was complete within a month. This action may be possible because of its effect against Candida albicans and probably other Candida spp. (antifungal activity). After complete healing also, the patients were advised to apply the drug once in the morning and once in the evening just before going to bed. They were also advised to avoid wetness and keep the affected area dry.. This stopped recurrence of the disease. This invention has been done for the first time and this drug is highly effective in the treatment and cure of Candidiasis which was not possible till date. No. - 5. Details regarding the search and/or examination report Review of literature on the present invention reveals that no attempt has been made by any other inventor on the control of Candidiasis by the ethanol extract of Myristica fragrans. To the best of our knowledge no patent has been filed on this material outside or within India, hence no question of any amendment on the specification of our invention submitted by us.REFERENCES 1. Grover, J. K., Khandkar, S., Vats, V., Dhunnoo, Y. and Das, D. 2002. Pharmacological studies onMyristicafragrans- antidiarrheal, hypnotic, analgesic and hemodynamic (bllod pressure) parameters. Methods Find Exp. Clin. Pharmacol.24: 675. 2. Hallstrom, A., Thuvander, A. 1997. Toxicological evaluations ofmyristicin. Nat.Toxins 5: 186-192. 3. Iwu, M. M. 1993. Handbook of African Medicinal Plants, 1st edn., pp 212-213, CRC Press, Florida. 4. Leung, K. H., Mihich, 1980. Prostaglandin modulation of development of cell mediated immunity in culture. Nature 288: 597-600. 5. Morita, T., Jinno, K., Kawagishi, H., Arimoto, Y., Suganuma, H., Inakuma, T., and Sugiyama, K. 2003. Hepatoprotective effect of myristicin from nutmeg (Myristica fragrans) of lipopolysaccharide/d-galacyosamine-induced liver injury. J. Agric. Food Chem.l2;51: 1560. 6. Olajide, O. A, Ajayi, F.F., Ekhelar, A. 1, Awe, S. O., Makinde, J. M. and AJada, R. 1999. Biological effects of Myristica fragrant (nutmeg) extract Phytother. Res. 13: 344. 7. Oliver-Bever, B. 1986. Medicinal plants in Tropical West Africa, pp 79-80. Cambridge University Press, London. 8. Park, S., Lee, D. K., Yang, C. H. 1998. Inhibition of tos-jun-DNA complex Formation by dihydroguaiaretic acid and in vitro cytotoxic effects on Cancer cells. Cancer Lett.15; 127:23. 9. Perez, C. and Anesini, C. 1994. Antibacterial activity of alimentary plants against Staphylococcus aureus growth. Amer. J. Clin. Med. 2: 169. 10. Ram, A., Lauria, P., Gupta, R. and Sharma, V. N. 1996. Hypolipidaemic effect of Myristica fragrans fruit extract in rabbits. J. Ethnopharmacol. 55: 49. 11. Sell, A. B. and Carlini, E. A. 1976. Anesthetic action of methyleugenol and other Eugenol derivatives. Pharmacology 14: 367. 12. Sharma, A. Mathur, R. and Dixit, V. P. 1995. Prevention of hypercholestero- lemia and atherosclerosclerosis in rabbits after supplementation of Myristica fragrans seed extract. Indian J. Physiol. Pharmacol. 39: 407. 13. Singh, A. and Rao, A. R. 1993. Modulatory effect of Areca nut on the action of mace {Myristica fragrans Hovtt) on the hepatic detoxification system in mice. Food Chem. Toxicol. 31:517. 14. Van Gils, C. and Cox, P. A. 1994. Ethnobotany of nutmeg in the spice islands. J. Ethnopharmacol. 42: 117. REFERENCES 1. Grover, J. K., Khandkar, S., Vats, V., Dhunnoo, Y. and Das, D. 2002. Pharmacological studies on Myristica fragrans- antidiarrheal, hypnotic, analgesic and hemodynamic (bllod pressure) parameters. Methods Find Exp. Clin. Pharmacol.24: 675. 2. Hallstrom, A., Thuvander, A. 1997. Toxicological evaluations of myristicin. Nat.Toxins 5: 186-192. 3. Iwu, M. M. 1993. Handbook of African Medicinal Plants, 1st edn., pp 212-213, CRC Press, Florida. 4. Leung, K. H., Mihich, 1980. Prostaglandin modulation of development of cell mediated immunity in culture. Nature 288: 597-600. 5. Morita, T., Jinno, K., Kawagishi, H., Arimoto, Y., Suganuma, H., Inakuma, T., and Sugiyama, K. 2003. Hepatoprotective effect of myristicin from nutmeg (Myristica fragrans) of lipopolysaccharide/d-galacyosamine-induced liver injury. J. Agric. Food Chem.l2;51: 1560. 6. Olajide, O. A, Ajayi, F.F., Ekhelar, A. I., Awe, S. O., Makinde, J. M. and Alada, R. 1999. Biological effects of Myristica fragrans (nutmeg) extract Phytother. Res. 13: 344. 7. Oliver-Sever, B. 1986. Medicinal plants in Tropical West Africa, pp 79-80. Cambridge University Press, London. 8. Park, S., Lee, D. K., Yang, C. H. 1998. Inhibition of tos-jun-DNA complex Formation by dihydroguaiaretic acid and in vitro cytotoxic effects on Cancer cells. Cancer Lett. 15; 127:23. 9. Perez, C. and Anesini, C. 1994. Antibacterial activity of alimentary plants against Staphylococcus aureus growth. Amer. J. Clin. Med. 2: 169. 10. Ram, A., Lauria, P., Gupta, R. and Sharma, V. N. 1996. Hypolipidaemic effect of Myristica fragrans fruit extract in rabbits. J. Ethnopharmacol. 55: 49. 11. Sell, A. B. and Carlini, E. A. 1976. Anesthetic action of methyleugenol and other Eugenol derivatives. Pharmacology 14: 367. 12. Sharma, A. Mathur, R. and Dixit, V. P. 1995. Prevention of hypercholestero- lemia and atherosclerosclerosis in rabbits after supplementation of Myristica fragrans seed extract. Indian J. Physiol. Pharmacol. 39: 407. 13. Singh, A. and Rao, A. R. 1993. Modulatory effect of Areca nut on the action of mace (Myristica fragrans Hovtt) on the hepatic detoxification system in mice. Food Chem. Toxicol. 31: 517. 14. Van Gils, C. and Cox, P. A. 1994. Ethnobotany of nutmeg in the spice islands. J. Ethnopharmacol. 42:117.

Full Text

COMPLETE SPECIFICATION
Title of the invention: Control of Camlidiasis by ethanol extract of Myristica
fragrans Objectives of invention
The objective of this invention is to provide a drug, which can control Candidiasis in human beings and also its recurrence without any side effects.
Myristica fragrans (Nutmeg)
The principal aromatic constituent of the volatile oil of nutmeg, the dry ripe seeds of M. fragrans is myristicin or methoxysafrole. This yields 7-16% volatile oil which contains approximately 4-8% myristicin, the nutmeg contains approximately 1.3% myristicin. In addition to it, the seed contains many other compounds also (Leung, 1980).
The chemical name of myristicin is 1 -allyl-5-methoxy-3, 4 methylene-dioxybenzene; CAS No. 607-91-0; empirical formula: C11H12O3; molecular weight: 192-22 (Merck Index, 1989).
CANDIDIASIS
Candidiasis is a fungal infection caused by related group of yeasts which are normal saprophytic inhabitants of mucocutancous body surfaces. They overgrow and invade tissues when permitted by alterations in the host. The causative organism is
usually Candida albicans. Its manifestations may be localized to the skin or it may even be systemic and life threatening.
Candidiasis occurs under the circumstances when a person
1. Suffers from diseases that either involve the immune system primarily or
require therapies that adversely affect immune competence.
2. Treated intensively with antibacterial agents, glucocorticoids or use of oral
contraceptives.
3. Has been inoculated intravenously, thus bypassing normal defences.
Candidal infections have affinity for sites that are commonly wet and macerated
for example corners of the mouth, oral cavity, around nails, intertriginous areas, penis, scrotum, inner aspects of thighs etc. In infants, cutaneous infections occur primarily as diaper rashes. This disease is frequently accompanied by a marked inflammatory response.
Almost all cases of oropharyngeal, gastrointestinal, vaginal, systemic and cutaneous Candidiasis are caused by C. albicans.
TREATMENT
It involves removal of predisposing factors such as systemic antibiotic, glucocorticoids therapy, chronic wetness and the use of appropriate topical azoles and systemic fluconazole, if required in severe cases. The dose of fluconazole depends on the site and severity of the disease.
Description of invention
The dry mature seeds of Myrislica fragrans were powdered in a pastle and mortar to make powder. One kilogram of powder was immersed in 1.5 1 of ethanol in a conical flask and kept at room temperature (25 ±2° C) for a period of 10 days. The flask was shaken manually several times daily for 10 days. After this period the material was filtered through cotton and the filtrate was collected in glass beakers. The filtrate was left in the beaker for one week at the above temperature for evaporation of ethanol. At the end of one week, the pH of the filtrate was 5.5 and the optical density was 760 nm.. This ethanol extract (drug) was applied locally to those patients who were suffering from Candidiasis in intertriginous area, around nails and were resistant to topical and systemic antifugal therapy. With the traditional antifungal therapy, the disease used to subside for some time and then again recur with severe itching, erythema and maceration of the affected area which used to be hyperkeratitic with treatment but never disappear. The application of this drug in such cases stopped itching within 2 days. Within a week the erythema completely disappeared with brownish discoloration of the area and the thickness and diameter of the hyperkeratotic plaque started reducing. After approximately two weeks the plaque reduced by 40%. Within four weeks of application, the plaque was hardly visible. Within next few days, there was complete healing of the affected area. This effect of the present drug seems to be miracle for the treatment of Candidiasis because with the traditional local and systemic antifugal therapy the plaque do not respond though the itching and erythema of the disease respond. This persistent hyperkeratotic plaque becomes responsible for recurrence of the disease. Our drug heals the affected skin to such an extent that the plaques disappear completely and thus do not
leave any chance of recurrence of the disease on that site. This effective action of this drug provides us with a comprehensive alternative to the other established modes of treatment, by curing the disease.
Present national and international knowledge
Many scientists have reported different properties of the extracts of dry seeds of M.fragrans. Sell and Carlini (1976) reported anaesthetic effect of methyl-euganol and other euganol derivatives of the volatile oil fraction of M. fragrans. In Nigeria and Indonesia, the essential oil of nutmeg is used in rheumatism (Oliver-Berver, 1986, Gils and Cox, 1994), probably because the oil decreases the level of prostaglandins which are major mediators of the inflammatory action (Iwo, 1983). This is also a known chemopreventor of chemically induced carcinogenesis (Singh and Rao, 1993). Us cytotoxic effect on cancer cells in vitro has been reported by Park et al (1998). Its antibacterial activity against Staphylococcnx aurens has been reported by Parez and Anesini (1994). Sharma et al. (1995) and Ram el al (1996) have reported prevention of hypercholesterolemia and atherosclerosis in rabbits after supplementation with seed extract. Olajide et al. (1999) described biological effects of chloroform extract of M. fragrans in male Swiss albino mice and male wistar rats as analgesic, anti-inflammatory and antithrombotic, whereas antidiarrheal activity of crude nutmeg solution as well as its petroleum ether extract has been reported by Grover et al. (2002). Myristicin which is the majopr component of essential oil of nutmeg is reported to posses extraordinarily potent hepatoprotective activity in mice against lipopolysaccharide (LPS) plus d-galactosamine (D-GalN) induced hepatotoxicity (Morita ci al. 2003). Though it has several other
properties its toxicological effects have not been proved as yet (Hall Strom and Thuvander, 1997).
There is also till today no report about the effect of ethanol extract of M.fragrans against Candidiasis of plaque type. Hence, this forms the first report in this direction establishing completely new invention which will prove to be the most important and effective treatment of Candidiasis without any side effects.
Present state of development
The ethanol extract of dry mature seeds of M.fragrans has been used in several patients suffering from Candidiasis with surprisingly complete healing effect.
Utility of the invention
The high rate of occurrence and recurrence of Cadidiasis is well known in spite of its treatment by traditional local and systemic antifugal drugs. The present drug starts acting by reducing the pruritis in Candidiasis. This is followed by reduction of erythema due to its anti-inflammatory action. Gradually this starts healing the macerated hyperkeratotic plaque and within a period of one month it completely heals leaving no chance for reoccurrence of the disease which is not possible by any other drug. It may also be possible because of its anti Candida albicans (antifungal) action. Since this drug is derived from an edible spice, which is commonly used in Indian kitchen, it may also be formulated for oral administration for the control of this disease. Till date its toxic effects have not been reported unless given in very high doses.
Novelty and nonobviousness
The effect of ethanol extract of M.fragmns is a completely new invention. It was not known earlier to this invention as is obvious from the review of literature till date regarding this property.
This drug is highly effective against Candidiasis. No other drug is known to treat this disease completely once it occurs. Hence, this drug seems to be much better and complete option for the treatment and cure of this disease.
DESCRIPTION OF THE INVENTION
The dry mature seeds of Myristica fragrans were powdered in a pastle and mortar. One kilogram of this powder was immersed in 1.5 1 of ethanol in a conical flask and kept at room temperature (25 ±2° C) for a period of 10 days. During this period the flask was repeatedly shaken and then the material was filtered through cotton and the filtrate was collected in glass beakers. It was left open at room temperature for one week. The pH of the filtrate was measured which was 5.5 and the optical density was 760 nm.
Thus obtained ethanol extract (drug) was tested for the treatment of Candidiasis also in those patients who had local and systemic antifungal therapy. It was gently applied to the affected areas thrice a day in both the groups. First application of the drug was soothing. Within two days the itching stopped completely. Within a week the erythema completely disappeared and the hyperkeratotic plaque started healing which was complete within a month. This action may be possible because of its effect against Candida albicans and probably other Candida spp. (antifungal activity). After complete healing also, the patients were advised to apply the drug once in the morning and once in the evening just before going to bed. They were also advised to avoid wetness and keep the affected area dry.. This stopped recurrence of the disease. This invention has been done for the first time and this drug is highly effective in the treatment and cure of Candidiasis which was not possible till date.
No. - 5. Details regarding the search and/or examination report
Review of literature on the present invention reveals that no attempt has been made by any other inventor on the control of Candidiasis by the ethanol extract of Myristica fragrans. To the best of our knowledge no patent has been filed on this material outside or within India, hence no question of any amendment on the specification of our invention submitted by us.REFERENCES
1. Grover, J. K., Khandkar, S., Vats, V., Dhunnoo, Y. and Das, D. 2002. Pharmacological
studies onMyristicafragrans- antidiarrheal, hypnotic, analgesic and hemodynamic (bllod
pressure) parameters. Methods Find Exp. Clin. Pharmacol.24: 675.
2. Hallstrom, A., Thuvander, A. 1997. Toxicological evaluations ofmyristicin. Nat.Toxins 5:
186-192.
3. Iwu, M. M. 1993. Handbook of African Medicinal Plants, 1st edn., pp 212-213, CRC
Press, Florida.
4. Leung, K. H., Mihich, 1980. Prostaglandin modulation of development of cell mediated
immunity in culture. Nature 288: 597-600.
5. Morita, T., Jinno, K., Kawagishi, H., Arimoto, Y., Suganuma, H., Inakuma, T., and
Sugiyama, K. 2003. Hepatoprotective effect of myristicin from nutmeg (Myristica
fragrans) of lipopolysaccharide/d-galacyosamine-induced liver injury. J. Agric. Food Chem.l2;51: 1560.
6. Olajide, O. A, Ajayi, F.F., Ekhelar, A. 1, Awe, S. O., Makinde, J. M. and AJada, R.
1999. Biological effects of Myristica fragrant (nutmeg) extract Phytother. Res. 13: 344.
7. Oliver-Bever, B. 1986. Medicinal plants in Tropical West Africa, pp 79-80. Cambridge
University Press, London.
8. Park, S., Lee, D. K., Yang, C. H. 1998. Inhibition of tos-jun-DNA complex
Formation by dihydroguaiaretic acid and in vitro cytotoxic effects on Cancer cells. Cancer
Lett.15; 127:23.
9. Perez, C. and Anesini, C. 1994. Antibacterial activity of alimentary plants against
Staphylococcus aureus growth. Amer. J. Clin. Med. 2: 169.
10. Ram, A., Lauria, P., Gupta, R. and Sharma, V. N. 1996. Hypolipidaemic effect of
Myristica fragrans fruit extract in rabbits. J. Ethnopharmacol. 55: 49.
11. Sell, A. B. and Carlini, E. A. 1976. Anesthetic action of methyleugenol and other
Eugenol derivatives. Pharmacology 14: 367.
12. Sharma, A. Mathur, R. and Dixit, V. P. 1995. Prevention of hypercholestero-
lemia and atherosclerosclerosis in rabbits after supplementation of Myristica fragrans
seed extract. Indian J. Physiol. Pharmacol. 39: 407.
13. Singh, A. and Rao, A. R. 1993. Modulatory effect of Areca nut on the action of
mace {Myristica fragrans Hovtt) on the hepatic detoxification system in
mice. Food Chem. Toxicol. 31:517.
14. Van Gils, C. and Cox, P. A. 1994. Ethnobotany of nutmeg in the spice islands. J.
Ethnopharmacol. 42: 117.

REFERENCES
1. Grover, J. K., Khandkar, S., Vats, V., Dhunnoo, Y. and Das, D. 2002. Pharmacological
studies on Myristica fragrans- antidiarrheal, hypnotic, analgesic and hemodynamic (bllod
pressure) parameters. Methods Find Exp. Clin. Pharmacol.24: 675.
2. Hallstrom, A., Thuvander, A. 1997. Toxicological evaluations of myristicin. Nat.Toxins 5:
186-192.
3. Iwu, M. M. 1993. Handbook of African Medicinal Plants, 1st edn., pp 212-213, CRC
Press, Florida.
4. Leung, K. H., Mihich, 1980. Prostaglandin modulation of development of cell mediated
immunity in culture. Nature 288: 597-600.
5. Morita, T., Jinno, K., Kawagishi, H., Arimoto, Y., Suganuma, H., Inakuma, T., and
Sugiyama, K. 2003. Hepatoprotective effect of myristicin from nutmeg (Myristica
fragrans) of lipopolysaccharide/d-galacyosamine-induced liver injury. J. Agric. Food Chem.l2;51: 1560.
6. Olajide, O. A, Ajayi, F.F., Ekhelar, A. I., Awe, S. O., Makinde, J. M. and Alada, R.
1999. Biological effects of Myristica fragrans (nutmeg) extract Phytother. Res. 13: 344.
7. Oliver-Sever, B. 1986. Medicinal plants in Tropical West Africa, pp 79-80. Cambridge
University Press, London.
8. Park, S., Lee, D. K., Yang, C. H. 1998. Inhibition of tos-jun-DNA complex
Formation by dihydroguaiaretic acid and in vitro cytotoxic effects on Cancer cells. Cancer
Lett. 15; 127:23.
9. Perez, C. and Anesini, C. 1994. Antibacterial activity of alimentary plants against
Staphylococcus aureus growth. Amer. J. Clin. Med. 2: 169.
10. Ram, A., Lauria, P., Gupta, R. and Sharma, V. N. 1996. Hypolipidaemic effect of
Myristica fragrans fruit extract in rabbits. J. Ethnopharmacol. 55: 49.
11. Sell, A. B. and Carlini, E. A. 1976. Anesthetic action of methyleugenol and other
Eugenol derivatives. Pharmacology 14: 367.
12. Sharma, A. Mathur, R. and Dixit, V. P. 1995. Prevention of hypercholestero-
lemia and atherosclerosclerosis in rabbits after supplementation of Myristica fragrans
seed extract. Indian J. Physiol. Pharmacol. 39: 407.
13. Singh, A. and Rao, A. R. 1993. Modulatory effect of Areca nut on the action of
mace (Myristica fragrans Hovtt) on the hepatic detoxification system in
mice. Food Chem. Toxicol. 31: 517.
14. Van Gils, C. and Cox, P. A. 1994. Ethnobotany of nutmeg in the spice islands. J.
Ethnopharmacol. 42:117.

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Patent Number

227364

Indian Patent Application Number

833/DEL/2004

PG Journal Number

04/2009

Publication Date

23-Jan-2009

Grant Date

07-Jan-2009

Date of Filing

06-May-2004

Name of Patentee

SINGH UDAI PRATAP

Applicant Address

A-2, NEW MEDICAL ENCLAVE, B.H.U., VARANASI-221005, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	SINGH UDAI PRATAP	A-2, NEW MEDICAL ENCLAVE, B.H.U., VARANASI-221005, INDIA.
2	SINGH RAVI VIKRAM	136, D SRI KRISHNAPURI, PATNA, BIHAR
3	SINGH MANDAVI	A-2, NEW MEDICAL ENCLAVE, B.H.U., VARANASI-221005, INDIA.

PCT International Classification Number

A61K36/882

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA