Title of Invention	"A PROCESS FOR THE PREPARATION OF A RADIO PROTECTIVE AND ANTI-CANCER HERBAL FORMULATION BASED ON RUBIA CORDIFOLIA AND SEMICORPUS ANACARDIUM"
Abstract	The process of present invention uses roots and rhizomes of Rubia cordifolia which are washed properly and extracted with water followed by filtration and centrifuging followed by extracting with ethanol. The crushed pieces of nuts of Semicarpus anacardium are washed properly and boiled with distilled water with mild heating. After concentration to semisolid state, it is again extracted with ethanol and purified on silica gel column by using hexane and finally with ethanol. The final ethanol extract of Rubia cordifolia and Semicarpus anacardium are mixed in certain ratio as disclosed herein to obtain the formulation of the present invention.

Title of Invention

"A PROCESS FOR THE PREPARATION OF A RADIO PROTECTIVE AND ANTI-CANCER HERBAL FORMULATION BASED ON RUBIA CORDIFOLIA AND SEMICORPUS ANACARDIUM"

Abstract

The process of present invention uses roots and rhizomes of Rubia cordifolia which are washed properly and extracted with water followed by filtration and centrifuging followed by extracting with ethanol. The crushed pieces of nuts of Semicarpus anacardium are washed properly and boiled with distilled water with mild heating. After concentration to semisolid state, it is again extracted with ethanol and purified on silica gel column by using hexane and finally with ethanol. The final ethanol extract of Rubia cordifolia and Semicarpus anacardium are mixed in certain ratio as disclosed herein to obtain the formulation of the present invention.

Full Text	FIELD OF INVENTION This invention relates a process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium. PRIOR ART Chemotherapy involving use of specific drugs and radiotherapy involving use of radiation are two established methods to stop uncontrolled growth of cancer cells. In both the cases there is damage to DNA leading to cell cycle arrest which is referred to as 'induction of apoptosis'. The main drawback of the above therapies is that these therapies are harmful to normal cells with the result that patients become weak and lose their immune defense system. There are two groups of medicines, which are used in cancer treatment. On is like Cicplatin, which forms the adduct with the DNA. Such medicines are successful as chemotherapeautic agents. Other group of anti cancer agents are those which inhibit the angeogenesis or inhinit the induciple cyclo-oxygenase 2 or induce the process of apoptosis by releasing the cytochrome C from mitochondria, thereby inducing the cellular capases. Some of chemical radioprotectors knoAvn in the art are cysteamine, Zn desferrioxamine, 2 aminopropyl-amino-ethane thiol, 2 mercaptopropionyl glycine (MPG), ellagic acid, bixin, melation, catecholestrogens, chlorogenic acid, 2 hydroxyestradiol, trimeprazin etc. The limitations of using these chemicals is that compromise normal physiological process. Yet another limitation of these chemicals is that these chemicals necessitate taking several medicines at a time. Several herbal compositions are being investigated for prevention of cancer growth and several pure phytochemicals have been isolated from herbs such as vincriestine from Vinca rosia, taxol from Taxus bacata. Some of these compounds have been synthesised and are also being modified by addition or deletion of some functional group on the parent skeletal group to make them function better. Some medicinal plants have shown radioprotection property. A few examples are Ocimum sanctum, Lycum Chinese, Withenia somnifera. Podophyllum hexandrum, Acanthopanax senticosus harms and curcumin. OBJECTS OF THE INVENTION. The primary object of the present invention is to propose a herbal formulation based on roots and rhizome of Rubia cordifolia and nuts of Semi carpus anacardium, which besides other possible applications, is specifically useful as an anticancer as well as radio protective agent. Another object of the present invention is to propose a herbal formulation which has antioxidant property and metal chelation property which help in better management of cancer patients. Yet another object of the present invention is to propose a herbal formulation based on combination of two herbs which is less toxic as compared to Semicarpus anacardium when taken alone. STATEMENT OF INVENTION According to this invention there is provide a process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium, comprising steps of :- (a) preparing plant extract front the washed roots and rhizomes of Rubia cordifolia, using water and taking water and plant material in the ratio of 16:1 (v/w), concentrating to one-fourth of its volume, filtering, centrifuging and concentrating the supematent liquid to semi-solid state followed by extraction with ethanol, obtaining fraction-I; (b) preparing plant extract from nuts of Semicarpus anacardium by crushing the said nuts into small pieces, mixing with dry finely powdered bricks washed with water and boiled with distilled water for 6-10hours with slow heating, filtering, concentrating to semi-solid followed by extraction with ethanol, purifying the ethanol fraction on silica gel column by using hexane and finally with ethanol and making ethanol fraction free from solvent, obtaining fraction-II; (c) mixing the said fraction-I as obtained by step (a) with fraction -II as obtained by step (b) in the ratio varying from 9.9:0.1 to 7:3, obtaining the desired herbal formulation. In accordance with the present invention, there is provided a herbal formulation which is based on two medicinal plants namely Rubia cordifolia (RC) and Semicarpus amacardium (SA). The herbal formulation is relatively less toxic as compared to Semicarpus amacardium if taken alone. The herbal formulation has both anticancer as well as radio protective property. As recommended by WHO, the process uses primary extraction of plants with water instead of using organic solvents for primary extraction. The organic solvents used in the processes of known art for extracting form plants, lead to several phytochemicals, which may be harmful to human beings. In the process of present invention, only the phytochemicals which are time tested for use by human beings and which can be extracted in water only are used and not the organic solvent extracted hytochemicals. DESCRIPTION OF FIGURES The results of evaluation of the formulation of the present invention for their radio protective property and anticancer property are illustrated with the help of accompanying figures, wherein- Fig 1 (a) shows the acute toxicity (72 hours) results with single dose in the range of 460 to 1500 mg/kg body weight Fig 1(b) shows effective time window for single dose of 460 kg/kg body weight, intraperitonial, before irradiation Fig 1(c) : shows survival of irradiated mice (single dose, IP, mg/kg bw) Fig 2 (a) shows liver lipid peroxidation - protective effect of formulation on radiation induced changes Fig 2 (b) shows mitotic inhibition in bone marrow - protective effect of Formulation Fig 3 (a) shows micronuclie formulation Fig 3 (b) shows colony forming units in spleen (CFU) Fig 4 (a) shows effect of formulation (200 mg/kg bw) on survival after 70 days treatment of DMBA (200mg/rat, IP) induced breast cancer carcinoma in albino rats Fig 4 (b) shows the effect of formulation (200 mg/kg bw) after 70 days Treatment on tumour volume for DMBA (200 mg/rat, IP), treated albino rats Fig 5(a) shows the preventive effect of formulation on Duct Carcinoma induced by DMBA Fig 5(b) shows the proliferative benign duct and fibrosis without tumour (magnification 150x, H&E) DESCRIPTION OF PROCESS According to the present invention, the process of preparation of the radio protective and anticancer formulation comprises of the following steps;-(a) Preparation of Extract of Rubia cordifolia The roots and rhizomes of Rubia cordifolia are taken and washed properly and extracted with water, taking water and plant roots and rhizomes, preferably in volume by weight ratio of 16:1. When water becomes about one-fourth of its volume, it is filtered and centrifuged. The supematent liquid is concentrated to semi-solid state and then extracted with ethanol. (b) Preparation of Extract of Semicarpus anacardium Nuts of Semicarpus anacardium are first crushed into small pieces and mixed with dry, finely powdered bricks. After 3-4 days, these nuts are washed with water properly and boiled with distilled water for 6-10 hours on slow heating device. It is then filtered and concentrated to semi-solid state followed by extraction with ethanol. The ethanol fraction is then purified on silica gel column by using hexane and finally with ethanol. The ethanol fi-action is made solvent free. (c) Preparation of Herbal Formulation The herbal formulation of the present invention is prepared by mixing the final ethanol extract of Rubia cordifolia (RC) obtained by step (a) with the final ethanol extract of Semicarpus anacardium (SA) in the preferred ratio (RC:SA) in the range between 9.9:0.1 to 7:3. It is to be noted that the composition is not a mere admixture with aggregation of properties. Each of the constituent does not show both radioprotective and anticancer properties whereas the composition shows both the radioprotective and hticancer property. Individually Semicarpus anacardium (SA) is relatively more beloxic where the combination upto 460 mg/kg body weight is not toxic even with a single dose,IP. The invention will now be illustrated with a working example which is intended to be a typical example to illustrate the working of the invention and is not intended to be taken restrictively to imply any limitation on the scope of the present invention. WORKING EXAMPLE RC Extract: 100 g of the roots and rhizomes of Rubia cordifolia are taken and washed properly and extracted with 1600 ml of water (16 part water; 1 part of drug). When water becomes about one-fourth of its volume, it is filtered and centrifuged. The supernatant liquid was concentrated to semi-solid state and then extracted with ethanol. The final yield was 14 g. It was dried and powdered. SA Extract: l00g of Nuts of Semicarpus anacardium was first crushed into small pieces and mixed with dry finely powdered bricks. .Aiter 3-4 days, these nuts were washed with water properly and boiled with distilled water for 6-10 hours on slow-heating device. Thereafier it was filtered and concentrated to semi-solid state. It was then filtered and concentrated to semi-solid state followed by extraction with ethanol. The ethanol fraction was purified on silica gel column by using hexane and finally with ethanol. The ethanol fraction is made solvent free. The yield was 2.4 g. It was dried and powdered. 10 g of RC extract was. properly mixed with the 0.2 g of SA extract. This mixture was used for all the experiments. METHOD OF USE For patients of Radiotherapy, the patient is orally given this medicine 3 times a day The morning dose is given 90 minutes before the Radiation. Dose of drug will be 200 mg in 3 divided doses. Detailed Experimental Studies and Their Results (a) Radio protective Property: Different doses of Co60 Radiation was given to rats. To these rats, different doses of herbal formulation were given at different time periods, prior to radiation. Afler radiation, animals were sacrificed and Mitotic count, Micronuclei CFU Assay (Colony Forming units in spleen). Liver lipid per-oxidation. Survival Assay, were examined. Results are given in Fig 1(a) to l(c ), Fig 2(a) to 2 (b) and Fig 3(a) to 3(b). It is observed that the mice which are pre-treated with formulation 1.5 hours before irradiation, showed significant protection against radiation induced lipid per-oxidation in the liver. It also inhibited the degree of radiation induced suppression of the dividing cells in the bone marrow (Mitotic index) and in spleens (eCFU). Further it was observed that this drug significantly protected the DNA against radiation induced damage as the number of cells having micronuclei were significantly less in the drug treated animals. (b) Anti Cancer Property : DMBA (7,12 Dimethyl benzanthracene) 20 mg/rat was injected to the female rats to produce breast carcinoma. The animals were divided into two groups. One group was given drug vehicle and the other group was regularly given formulation in the dose of 200 mg/kg BW orally (daily one dose in the morning for 10 weeks). Animals were sacrificed and the histopathological studies were made. Results are given in Fig 4(a), 4(b), 5(a) and 5(b). There was significant protection against the DMBA (7,12 Dimethyl benzanthracene) induced breast cancer carcinoma as the drug treated animals showed significant increase in the survival rate of animals. It was in the range of 66% to 100%. The tumour volume was reduced to 50% when compared to the experimental control. The AgNOR dots in the nucleus of the treated animals were also significantly less in the range of 35%-50%, (c) Toxicity Study : To study the acute toxicity (72 hours) single dose of Drug was given intraperitonially in different doses in the range of 460 to 1500 mg/kg body weight. After 72 hours, rate of mortality was determined. Safe dose was found to be 460 mg/kg body weight which was used in all the experiments. Fig 1 (a) refers. -(0-For chronic study, the animals were subject to oral drug administration in the dose range of 180-500 mg/kg body weight. Single dose was given every day up to 30 days. Finally the animals were sacrificed and following parameters were carried out on Serum GOT, Serum GPT, LIVER LPO, Liver GSH. Results are given in Table 1 below. In the acute toxicity study, the drug was completely safe on low doses. No mortality was noted after 72 hours of intraperitonial injection of single dose of drug in the range of 220-450 mg/kg body weight. In the case of chronic toxicity, one month oral treatment did not show any change in the hepatocellular enzymes namely SGOT, SGPT and alkaline phosphatase. TABLE-I CHRONIC TOXICITY - ONE MONTH ORAL TREATMENT (Table Removed) Values are mean ± SD of 6 different experiments. It is to be understood that the formulation of the present invention and process for preparation thereof is susceptible to modifications, changes and adaptations by those skilled in the art. Such modifications, changes, adaptations are intended to be within the scope of the present invention which is further set forth under the following claims:- I CLAIM; 1- A process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium, comprising steps of :- (a) preparing plant extract front the washed roots and rhizomes of Rubia cordifolia, using water and taking water and plant material in the ratio of 16:1 (v/w), concentrating to one-fourth of its volume, filtering, centrifuging and concentrating the supematent liquid to semi-solid state followed by extraction with ethanol, obtaining fraction-I; (b) preparing plant extract from nuts of Semicorpus anacardium by crushing the said nuts into small pieces, mixing with dry finely powdered bricks washed with water and boiled with distilled water for 6-l0hours with slow heating, filtering, concentrating to semi-solid followed by extraction with ethanol, purifying the ethanol fraction on silica gel column by using hexane and finally with ethanol and making ethanol fraction free from solvent, obtaining fraction-II; (c) mixing the said fraction-I as obtained by step (a) with fraction -II as obtained by step (b) in the ratio varying from 9.9:0.1 to 7:3, obtiiining the desired hebal formulation. 2. A process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium as substantially described and illustrated herein.

Full Text

FIELD OF INVENTION
This invention relates a process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium.
PRIOR ART
Chemotherapy involving use of specific drugs and radiotherapy involving use of radiation are two established methods to stop uncontrolled growth of cancer cells. In both the cases there is damage to DNA leading to cell cycle arrest which is referred to as 'induction of apoptosis'.
The main drawback of the above therapies is that these therapies are harmful to normal cells with the result that patients become weak and lose their immune defense system.
There are two groups of medicines, which are used in cancer treatment. On is like Cicplatin, which forms the adduct with the DNA. Such medicines are successful as chemotherapeautic agents. Other group of anti cancer agents are those which inhibit the angeogenesis or inhinit the induciple cyclo-oxygenase 2 or induce the process of apoptosis by releasing the cytochrome C from mitochondria, thereby inducing the cellular capases.
Some of chemical radioprotectors knoAvn in the art are cysteamine, Zn desferrioxamine, 2 aminopropyl-amino-ethane thiol, 2 mercaptopropionyl glycine (MPG), ellagic acid, bixin, melation, catecholestrogens, chlorogenic acid, 2 hydroxyestradiol, trimeprazin etc.
The limitations of using these chemicals is that compromise normal physiological
process.
Yet another limitation of these chemicals is that these chemicals necessitate taking several medicines at a time.
Several herbal compositions are being investigated for prevention of cancer growth and several pure phytochemicals have been isolated from herbs such as vincriestine from Vinca rosia, taxol from Taxus bacata. Some of these compounds have been synthesised and are also being modified by addition or deletion of some functional group on the parent skeletal group to make them function better. Some medicinal plants have shown radioprotection property. A few examples are Ocimum sanctum, Lycum Chinese, Withenia somnifera. Podophyllum hexandrum, Acanthopanax senticosus harms and curcumin.
OBJECTS OF THE INVENTION.
The primary object of the present invention is to propose a herbal formulation based on roots and rhizome of Rubia cordifolia and nuts of Semi carpus anacardium, which besides other possible applications, is specifically useful as an anticancer as well as radio protective agent.
Another object of the present invention is to propose a herbal formulation which has antioxidant property and metal chelation property which help in better management of cancer patients.
Yet another object of the present invention is to propose a herbal formulation based on combination of two herbs which is less toxic as compared to Semicarpus anacardium when taken alone.
STATEMENT OF INVENTION
According to this invention there is provide a process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium, comprising steps of :-
(a) preparing plant extract front the washed roots and rhizomes of Rubia cordifolia, using water and taking water and plant material in the ratio of 16:1 (v/w), concentrating to one-fourth of its volume, filtering, centrifuging and concentrating the supematent liquid to semi-solid state followed by extraction with ethanol, obtaining fraction-I;
(b) preparing plant extract from nuts of Semicarpus anacardium by crushing the said nuts into small pieces, mixing with dry finely powdered bricks washed with water and boiled with distilled water for 6-10hours with slow heating, filtering, concentrating to semi-solid followed by extraction with ethanol, purifying the ethanol fraction on silica gel column by using hexane and finally with ethanol and making ethanol fraction free from solvent, obtaining fraction-II;
(c) mixing the said fraction-I as obtained by step (a) with fraction -II as obtained by step (b) in the ratio varying from 9.9:0.1 to 7:3, obtaining the desired herbal formulation.
In accordance with the present invention, there is provided a herbal formulation which is based on two medicinal plants namely Rubia cordifolia (RC) and Semicarpus amacardium (SA). The herbal formulation is relatively less toxic as compared to Semicarpus amacardium if taken alone. The herbal formulation has both anticancer as well as radio protective property. As recommended by WHO, the process uses primary extraction of plants with water instead of using organic solvents for primary extraction. The organic solvents used in the processes of known art for extracting form plants, lead to several phytochemicals, which may be harmful to human beings. In the process of present invention, only the phytochemicals which are time tested for use by human beings and which can be extracted in water only are used and not the organic solvent extracted hytochemicals.
DESCRIPTION OF FIGURES
The results of evaluation of the formulation of the present invention for their radio protective property and anticancer property are illustrated with the help of accompanying figures, wherein-
Fig 1 (a) shows the acute toxicity (72 hours) results with single
dose in the range of 460 to 1500 mg/kg body weight
Fig 1(b) shows effective time window for single dose of 460
kg/kg body weight, intraperitonial, before irradiation
Fig 1(c) : shows survival of irradiated mice (single dose, IP,
mg/kg bw)
Fig 2 (a) shows liver lipid peroxidation - protective effect of
formulation on radiation induced changes
Fig 2 (b) shows mitotic inhibition in bone marrow - protective
effect of Formulation
Fig 3 (a) shows micronuclie formulation
Fig 3 (b) shows colony forming units in spleen (CFU)
Fig 4 (a) shows effect of formulation (200 mg/kg bw) on
survival after 70 days treatment of DMBA
(200mg/rat, IP) induced breast cancer carcinoma in
albino rats
Fig 4 (b) shows the effect of formulation (200 mg/kg bw) after
70 days
Treatment on tumour volume for DMBA (200 mg/rat, IP), treated albino rats
Fig 5(a) shows the preventive effect of formulation on Duct Carcinoma
induced by DMBA
Fig 5(b) shows the proliferative benign duct and fibrosis without tumour
(magnification 150x, H&E)
DESCRIPTION OF PROCESS
According to the present invention, the process of preparation of the radio protective and anticancer formulation comprises of the following steps;-(a) Preparation of Extract of Rubia cordifolia
The roots and rhizomes of Rubia cordifolia are taken and washed properly and extracted with water, taking water and plant roots and rhizomes, preferably in volume by weight ratio of 16:1. When water becomes about one-fourth of its volume, it is filtered and centrifuged. The supematent liquid is concentrated to semi-solid state and then extracted with ethanol.
(b) Preparation of Extract of Semicarpus anacardium
Nuts of Semicarpus anacardium are first crushed into small pieces and mixed with dry, finely powdered bricks. After 3-4 days, these nuts are washed with water properly and boiled with distilled water for 6-10 hours on slow heating device. It is then filtered and concentrated to semi-solid state followed by extraction with ethanol. The ethanol fraction is then purified on silica gel column by using hexane and finally with ethanol. The ethanol fi-action is made solvent free.
(c) Preparation of Herbal Formulation
The herbal formulation of the present invention is prepared by mixing the final ethanol extract of Rubia cordifolia (RC) obtained by step (a) with the final ethanol extract of Semicarpus anacardium (SA) in the preferred ratio (RC:SA) in the range between 9.9:0.1 to 7:3.
It is to be noted that the composition is not a mere admixture with aggregation of properties. Each of the constituent does not show both radioprotective and anticancer properties whereas the composition shows both the radioprotective and
hticancer property. Individually Semicarpus anacardium (SA) is relatively more beloxic where the combination upto 460 mg/kg body weight is not toxic even with a
single dose,IP.
The invention will now be illustrated with a working example which is intended to be a typical example to illustrate the working of the invention and is not intended to be taken restrictively to imply any limitation on the scope of the present invention.
WORKING EXAMPLE
RC Extract: 100 g of the roots and rhizomes of Rubia cordifolia are taken and washed properly and extracted with 1600 ml of water (16 part water; 1 part of drug). When water becomes about one-fourth of its volume, it is filtered and centrifuged. The supernatant liquid was concentrated to semi-solid state and then extracted with ethanol. The final yield was 14 g. It was dried and powdered.
SA Extract: l00g of Nuts of Semicarpus anacardium was first crushed into small pieces and mixed with dry finely powdered bricks. .Aiter 3-4 days, these nuts were washed with water properly and boiled with distilled water for 6-10 hours on slow-heating device. Thereafier it was filtered and concentrated to semi-solid state. It was then filtered and concentrated to semi-solid state followed by extraction with ethanol. The ethanol fraction was purified on silica gel column by using hexane and finally with ethanol. The ethanol fraction is made solvent free. The yield was 2.4 g. It was dried and powdered.
10 g of RC extract was. properly mixed with the 0.2 g of SA extract. This mixture was used for all the experiments.
METHOD OF USE
For patients of Radiotherapy, the patient is orally given this medicine 3 times a day The morning dose is given 90 minutes before the Radiation. Dose of drug will be 200 mg in 3 divided doses.
Detailed Experimental Studies and Their Results
(a) Radio protective Property: Different doses of Co60 Radiation was given to
rats. To these rats, different doses of herbal formulation were given at different time
periods, prior to radiation. Afler radiation, animals were sacrificed and Mitotic count,
Micronuclei CFU Assay (Colony Forming units in spleen). Liver lipid per-oxidation.
Survival Assay, were examined.
Results are given in Fig 1(a) to l(c ), Fig 2(a) to 2 (b) and Fig 3(a) to 3(b). It is observed that the mice which are pre-treated with formulation 1.5 hours before irradiation, showed significant protection against radiation induced lipid per-oxidation in the liver. It also inhibited the degree of radiation induced suppression of the dividing cells in the bone marrow (Mitotic index) and in spleens (eCFU). Further it was observed that this drug significantly protected the DNA against radiation induced damage as the number of cells having micronuclei were significantly less in the drug treated animals.
(b) Anti Cancer Property : DMBA (7,12 Dimethyl benzanthracene) 20 mg/rat was injected to the female rats to produce breast carcinoma. The animals were divided into two groups. One group was given drug vehicle and the other group was regularly given formulation in the dose of 200 mg/kg BW orally (daily one dose in the morning for 10 weeks). Animals were sacrificed and the histopathological studies were made. Results are given in Fig 4(a), 4(b), 5(a) and 5(b). There was significant protection against the DMBA (7,12 Dimethyl benzanthracene) induced breast cancer carcinoma as the drug treated animals showed significant increase in the survival rate of animals. It was in the range of 66% to 100%. The tumour volume was reduced to 50% when compared to the experimental control. The AgNOR dots in the nucleus of the treated animals were also significantly less in the range of 35%-50%,
(c) Toxicity Study : To study the acute toxicity (72 hours) single dose of Drug was given intraperitonially in different doses in the range of 460 to 1500 mg/kg body weight. After 72 hours, rate of mortality was determined. Safe dose was found to be 460 mg/kg body weight which was used in all the experiments. Fig 1 (a) refers.
-(0-For chronic study, the animals were subject to oral drug administration in the dose range of 180-500 mg/kg body weight. Single dose was given every day up to 30 days. Finally the animals were sacrificed and following parameters were carried out on Serum GOT, Serum GPT, LIVER LPO, Liver GSH.
Results are given in Table 1 below. In the acute toxicity study, the drug was completely safe on low doses. No mortality was noted after 72 hours of intraperitonial injection of single dose of drug in the range of 220-450 mg/kg body weight. In the case of chronic toxicity, one month oral treatment did not show any change in the hepatocellular enzymes namely SGOT, SGPT and alkaline phosphatase.
TABLE-I CHRONIC TOXICITY - ONE MONTH ORAL TREATMENT

(Table Removed)
Values are mean ± SD of 6 different experiments.
It is to be understood that the formulation of the present invention and process for preparation thereof is susceptible to modifications, changes and adaptations by those skilled in the art. Such modifications, changes, adaptations are intended to be within the scope of the present invention which is further set forth under the following claims:-

I CLAIM;
1- A process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium, comprising steps of :-
(a) preparing plant extract front the washed roots and rhizomes of Rubia cordifolia, using water and taking water and plant material in the ratio of 16:1 (v/w), concentrating to one-fourth of its volume, filtering, centrifuging and concentrating the supematent liquid to semi-solid state followed by extraction with ethanol, obtaining fraction-I;
(b) preparing plant extract from nuts of Semicorpus anacardium by crushing the said nuts into small pieces, mixing with dry finely powdered bricks washed with water and boiled with distilled water for 6-l0hours with slow heating, filtering, concentrating to semi-solid followed by extraction with ethanol, purifying the ethanol fraction on silica gel column by using hexane and finally with ethanol and making ethanol fraction free from solvent, obtaining fraction-II;
(c) mixing the said fraction-I as obtained by step (a) with fraction -II as obtained by step (b) in the ratio varying from 9.9:0.1 to 7:3, obtiiining the desired hebal formulation.
2. A process for preparation of a radioprotective and anticancer herbal formulation based on Rubia cordifolia and Semicorpus anacardium as substantially described and illustrated herein.

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Patent Number

195018

Indian Patent Application Number

661/DEL/2002

PG Journal Number

31/2009

Publication Date

31-Jul-2009

Grant Date

17-Mar-2006

Date of Filing

18-Jun-2002

Name of Patentee

ADDITIONAL DIRECTOR (IPR)

Applicant Address

DEFENCE RESEARCH & DEVELOPMENT ORGANISATION MINISTRY OF DEFENCE, GOVT OF INDIA, B-341, SENA BHAWAN, DHQ P.O. NEW DELHI-110011, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	YAMINI BHUSHAN TRIPATHI	DEPTT. OF MEDICINAL CHEMISTRY, INSTITUTE OF MEDICAL SCIENCES, BANARAS HINDU UNIVERSITY, INDIA.

PCT International Classification Number

A61K 31/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA