Indian Patents. 209018:A BONE FRACTURE HEALING EXTRACT FROM THE HERB ASYSTASIA GANGETICA AND A PROCESS FOR ITS PREPARATION

Title of Invention	A BONE FRACTURE HEALING EXTRACT FROM THE HERB ASYSTASIA GANGETICA AND A PROCESS FOR ITS PREPARATION
Abstract	This invention relates to an extract from an herbal plant having bone fracture healing properties. The invention particularly relates to an extract from the herb As~stasia gangetica This invention also relates to a process for the preparation of the above said extract having bone fracture healing properties from the herb As~stasia aanaetica.

Full Text	This invention relates to an extract from an herbal plant having bone fracture healing properties. The invention particularly relates to an extract from the herb Asvstasia gangetica (family: Acanthaceae). This invention also relates to a process for the preparation of the above said extract having bone fracture healing properties from the herb Asvstasia gangetica. The prolonged period that is generally required for healing of fractures is a matter of great concern and frustration both to the patient and to the orthopedic surgeon. Treatment of bone fracture often involves surgery, which may prove hazardous to very elderly patients. Even after treatment one can't be certain whether a particular fracture gets healed normally or results in delayed union or even non-union. Because of these uncertainties the patient may have to wait for a prolonged period before he can use the fractured limb properly. Besides applying plaster with or without surgery, there is no rational clinical treatment to hasten the healing process although vitamins and anabolic hormones can play a useful role especially when the patient is deficient in them. The main objective of the present invention is therefore to provide an extract from the herb Asvstasia gangetica having bone fracture healing properties. Another objective of the present invention is to provide a process for the preparation of an extract from the leaves /stems of the herb Asvstasia gangetica having bone fracture healing properties. The present invention provides a process for the preparation of a medicament from the leaves and stems of Asvstasia gangetica (family: Acanthaceae) by extraction with water or aqueous alcohol at controlled temperature. So far only one herb namely Cissus guadrangularis (family: Vitaceae) has been reported [K.N.Udupa, H.J.Arnikar, L.M.Singh - Experimental studies of the use of Cissus auandrangularis in healing fracture, Indian.J.Med.Sci, 1961,l5,551;K.N.Udupa and G.C.Prasad - Further Studies on the effect of Cissus quadrangularis in accelerated fracture healing, Indian J.Med.Res, 1964,52,1] to have bone fracture healing property. Chemical analysis of this herb revealed a high concentration of vitamin C besides carotene, calcium oxalate and a substance like "anabolic steroid'. It may be mentioned that calcium oxalate is an undesirable substance in many herbs, at times in high concentration. Excessive ingestion of calcium oxalate will lead to the complaint oxaluria. While vitamin C and carotene have nutritive effect, the nature and effect of the "anabolic steroid" is not known till now. Since ancient times the herb has been in use, especially by bone setters in villages for both external application as paste and as an internal medicine to be taken with milk. For scientific investigation an alcoholic extract of the herb was obtained by cold percolation method, drying it and then dissolving it in ethylene glycol and administering by intramuscular injection. This has hastened fracture healing by reducing the total convalescent period by 33% in the experimental animals. In subsequent animal experiments [K.N.Udupa and G.C. Prasad, Biomedical and Calcium'*^ studies on the effect of Cissus quadrangularis in fracture repair, Indian J.Med.Res, 1964, 52, 480; K.N.Udupa. G.C.Prasad and S.P.Sen, The effect of phytogenic anabolic steroid in the acceleration of fracture repair. Life science, 1965, 4, 317]. Both the aqueous alcoholic extract and a lipid soluble "phytogenic steroid" (anabolic steroid) were found to possess bone fracture healing property when given by intra peritoneal route. Both have shown toxicity when the dosage was increased. Till now neither the aqueous alcoholic extract nor the "phytogenic steroid" have been developed for clinical studies and therapeutic use. Except this herb no other herb has been investigated nor generally known to have bone fracture healing properties. Asystasia gangetica (Linn) (synonym: Asvstasia coromandeliana) family: Acanthaceae) is an erect unarmed glabrous herb, 50 to 100 cm.high, branches arising from the base, leaves orate, entire, glabrous, subacuminate 4 x 1.5 - 2.5 cm. Kacemes up to 30 cm. long; corolla white capsules enclosed within the persistent calyx teeth. It is an abundant weed during the rainy season forming large patches in open places. In Sanskrit it is called LAVANAVALLI. In medicinal plant literature [R.N.Chopra, S.L.Nayar and I.e. Chopra, Glossary of Indian Medicinal Plants, Publications and Information Directorate, C.S.I.R., New Delhi, p.30 (1956)]. It was mentioned that the juice of the plant is anthelmentic, given in rheumatism and swellings. The use of the leaves of the above mentioned herb for healing of fracture is practiced in Haryana. As per this practice the personnel who are beaten by police with lathis is asked to take the dried powder of the leaves of the said herb along with milk. It is the belief of the people in that State that by doing so the personnel inflicted with injuries to the bone as a result of such beatings get relief. Such a practice is not prevalent in other parts of the country. There is no published information about this practice based on the belief followed in Haryana. Another peculiarity of this herb is that it is not available throughout the year and throughout India, The plant grows only in rainy season and that too only in certain parts of India like Haryana and Andhra Pradesh. This means the herb is location and climate specific. Keeping in mind the folklore use of the leaves of the herb, and the importance of providing relief for the patients suffering from bone injuries, we took the initiative to study the medicinal properties of this herb in a systematic way. Our detailed studies on the herb revealed that in addition to the presence of active ingredients, responsible for imparting relief to the injuries inflicted to the bones, in the parts of the herb, especially the leaves, it also contains insoluble salts of calcium like calcium oxalate and toxic metals like lead, arsenic and mercury which are toxic and therefore injurious to health. Therefore by continuous intake of the raw leaves directly, though there is immediate relief of the injuries to the bones, due to the presence of above said toxic materials, which also simultaneously adversely affects the health of the patient in a longer run leading to other complications. These dangerous consequences are not understood or realized by those who are traditionally practicing the use of the said herb for healing bone injuries. Our systematic and scientific studies on the bone healing properties of the herb Asvstasia gangetica such as leaves, stems, flowers, roots, etc. Indicates that the parts of the herb are rich in minerals and organic constituents, which are responsible for imparting such properties to the herb. The main constituents which imparts the bone healing properties to the parts of the herb, particularly the leaves and the stems are found to be the salts of sodium, potassium, calcium (other than calcium oxalate), magnesium and zinc and organic constituents like sugars and polyphenols (flavonoids). These active ingredients are present in specific amounts. Hence the inventive step lies in identifying the main constituents for bone healing properties from the Asvstasia gangetica. Accordingly The main constituents which imparts the bone healing properties to the parts of the herb, particularly the leaves and the stems are found to be the salts of sodium, potassium, calcium (other than calcium oxalate), magnesium and zinc and organic constituents like sugars and polyphenols (flavonoids). These active ingredients are present in specific amounts. Therefore the main objective of the present invention is to provide an extract having bone fracture healing properties from the herb Asvstasia gangetica (family: Acanthaceae) Another objective of the present invention is to provide a process for the preparation of the extract having bone fracture healing properties from the herb Asvstasia gangetica (family: Acanthaceae) The principle by which the invention has been developed is based on the folklore use of the herb Asvstasia gangetica (familv: Acanthaceae) practiced in Haryana and by extracting the parts of the said herb employing a specific method which will extract only the organic and inorganic active ingredients wliicli do not undergo any deconiposition and avoiding extraction of any of the toxic ingredients present therein. Accordingly the present invention provides an extract from the parts of the herb, Asystasia gangetica (family: Acanthaceae) having bone fracture healing properties comprising I. Salts of sodium in the range of 0.40 to 2.4% by weight. II. Salts of potassium in the range of 3.5 to 6.1%. By weight. III. Salts of calcium (excluding calcium oxalate) in the range of 1.0 to 5.5%. By weight. IV. Salts of magnesium in the range of 0.12 to 2.5%. By weight. V. Salts of zinc in the range of 0.001 to 0.004% by weight. VI. Sugars in the range of 8,4 to 29.3%. By weight. VII. Polyphenols (flavonoids) in the range of 8.1 to 12.9 %. By weight. Vlil. Other unidentified nontoxic agents in the range of 78.48 to 41.3%. By weight The metals mentioned above are present in the form of their salts (other than oxalate) like chloride, sulphate, lactate, tartarate and citrate. They are estimated and expressed as metals throughout. The amount of the above mentioned active constituents vary depending upon the parts used for extraction. The salts of the metals may be present as chloride, sulphate, lactate, citrate, tartrate, etc. It is important to note that the extract does not contain calcium oxalate and other toxic metal salts of Arsenic, Lead, Mercury, and Cadmium etc., which are usually present in the parts of the herb. The sugars and flavonoids have not yet been identified. There is likely that some of these agents may be novel. Similarly work is in progress to identify the other unidentified non-toxic components. It may be possible that all or some of these agents may turn out to be new agents. The amount of the salt of sodium may preferably range from 0.6 to 2.1% by weight. The amount of the salt of potassium may preferably range from 3.7 to 5.8% by weight. The amount of the salt of calcium (other than toxic calcium salts) may preferably range from 1.4 to 5.1% by weight. The amount of the salt of magnesium may preferably range from 0.15 to 2.1% by weight. The amount of the salt of zinc may preferably range from 0.002 to 0.003% by weight. The amount of sugars may preferably range from 10 to 25% by weight. The amount of polyphenols (flavonoids) may preferably range from 9 to 11.5% by weight. According to another feature of the present invention there is provided a process for the preparation of the above-mentioned extract from the parts of the herb, Asvstasia qangetica (family Acanthaceae) having bone fracture healing properties comprising. I. Powdering the dried parts of the herb Asvstasia gangetica (family :Acanthaceae) II. Extracting the resulting powder with a polar solvent or a mixture of polar solvents at a temperature in the range of 35 to 45 0C using a water bath. III. Maintaining the temperature of the bath strictly at the above said temperature while stirring continuously for a period in the range of 2 to 6 hrs IV. Filtering the resulting extract to remove any suspended material V. Washing the residue with the same solvent or mixture of solvents used for the extraction. VI. Concentrating the resulting filtrate under reduced pressure by heating in a water bath maintaining the temperature in the range of 45 to 55°C and if desired. VII. Forming pills /tablets /capsules/liquid orals of the resulting product by conventional methods. In a preferred embodiment of the invention the polar solvent employed for extraction may be selected from distilled water, dematerialized water, alcohol or aqueous alcohol wherein the ratio of water to alcohol ranges from 5 to 20%. The sizes of the powder of the parts of the herb may range from 20 to 40 meshes. The resulting product which is in the form of soft or dry extract (containing 10-20% water) may be preserved by adding a preservative and can be made into Pills/tablets /capsules/liquid orals by the conventional meinoas. The invention is disclosed in detail in the examples given below which are provided by way of illustration only and therefore should not be construed to limit the scope of the invention. Example-1. The fresh leaves collected from Asvstasia gangetica herb are shade dried and powdered coarsely (20-40 mesh) in an electric grinder. The coarse powder (500 g) is taken into a 5 I. beaker and 3000 ml. dematerialized water is added. An electric paddle stirrer is introduced into the beaker. The beaker is kept in an electric water bath with thermostatic control. The water bath temperature is maintained in the range of 35-40'C with the help of thermostat. The extraction is carried out under continuous stirring for 4 hours. There after the solvent is filtered under suction and washed with 250 ml dematerialized water. The filtrate is once again filtered through a funnel containing a tight cotton plug to remove all suspended material. The clear filtrate is concentrated in a rotary evaporator over a water bath maintained at 45-50°C. The resulting product is a gummy solid soluble in water, yield 5.5%. The product contains salts of sodium 0.7%, potassium 3.5%, calcium (except calcium oxalate) 2.1%, magnesium 1.5 %, and zinc 0.002%, and sugars (reducing substances calculated as glucose) 8.4% and polyphenols (flavonoids) 8,1%. Pills for oral administration may be prepared after thoroughly mixing the extract with known pharmaceutical solids and /or liquid excipients and then rolling the mass into a rod of even thickness, cutting to the length required and rounding to obtain perfect spheres. Example-2 The stems of Asvstasia ganqetica are separated from leaves and shade dried. The dried stems are broken into small pieces and powdered in an electric grinder to a coarse powder (20-40 mesh). The coarse powder (450 g) is taken into a 5 I beaker and 2500 ml of distilled water is added. An electric paddle stirrer is introduced into the beaker. The beaker is kept in thermostat water bath maintained at 40-45°C. The extraction is carried out under continuous stirring for 3 hours. There after the solvent is filtered under reduced pressure and washed with 300 ml of distilled water. The filtrate is once again filtered through a large funnel using a fluted filter paper. The clear filtrate in concentrated in a film evaporator at 40"45°C to remove all water. The resulting product is a gummy solid soluble in water, yield 5.7%. The product contains salts of sodium 2.1%, potassium 5.3%, calcium (except calcium oxalate) 5.3%, magnesium 1.3 %, and zinc 0.003%, and sugars (reducing substances calculated as glucose) 9% and polyphenols (fiavonoids) 9.6%. Tablets for oral administration may be prepared from the extract by direct compression or wet granulation after dilution with microcrystalline cellulose, lactose, starch and other such materials used as diluents Example-3 The coarsely powdered dried leaf of Asvstasia gangetica (400 g) is treated with 10% aqueous alcohol (3500ml) (alcohol to water ratio 1:9 by volume) in a 5 I beaker. The beaker is immersed in an electric water bath having a thermostat maintained at 35-40°C. An electric stirrer is introduced into the beaker and the extraction is carried out with continuous stirring for 5 hours. Periodically small volumes (20-30 ml) of 10% alcohol are added to make good the loss of solvent by evaporation during extraction. Thereafter the solvent was filtered and the residue washed with 300 ml 10% aqueous alcohol. The clear filtrate was concentrated under reduced pressure at a temperature of 45-50°C to remove all alcohol. The aqueous concentrate is then freeze dried when the product was obtained as hygroscopic form which is freely soluble in water, yield 7.2%. The product contains salts of sodium 0.6%, potassium 5.9%, calcium (except calcium oxalate) 2.2%, magnesium 2.3 %, and zinc 0.002%, and sugars (reducing substances calculated as glucose) 13.4% and polyphenols (flavonoids) 11.5%. Soft gel capsules for oral administration may be prepared from the extract by dilution with suitable hydrophobic diluents such as corn oil or other vegetable oils and filled by known methods using rotary die process. Hard gelatin capsules may be prepared after adding moisture absorbing diluents like starch, anhydrous declaim phosphate and the like and filling the capsules by known methods. Example-4 The coarsely powdered dried stem of Asvstasia qanqetica (500 g) is treated with 15 % aqueous alcohol (3200 ml) (alcohol to water ratio 3: 17 by volume) in a two-necked 5 I round bottom flask fitted with a stirrer and a condenser. The round bottom flask is immersed in an electric water bath with a thermostat and the temperature maintained at 38-45°C. Extraction is carried out under continuous stirring for six hours. Thereafter the solvent was filtered through a thin bed of washed kieselguhr through a large Buchner funnel and the residue washed with 400 ml of 15 % aqueous alcohol. The clear filtrate was concentrated at 45-50°C in rotary evaporator to a dry extract containing ca.10% moisture. The resulting product is a gummy material freely soluble in water, yield 7.5%. The product contains salts of sodium 0.6%, potassium 5.3%, calcium (expect calcium oxalate) 1.9%, magnesium 1.0 %, and zinc 0.001%, and sugars (reducing substances calculated as glucose) 27.2% and polyphenols (flavonoids) 11.8 %. Oral liquid dosage form may be prepared by dissolving or dispersing ine extract in a suitable non-medicated syrup. The non-medicated syrup contains sweetening agent flavors, optionally containing colorants and ant microbial preservatives. Test results of the bone healing properties of the product obtained from examples 1 to 4 of the invention: The aqueous extracts obtained by examples 1 and 2 or hydroalcoholic extracts obtained by examples 3 and 4 when administered orally as aqueous solution/suspension to rats in which fracture was produced artificially in the right tibia and fibula showed enhanced bone healing properties compared to control group after one week. After two weeks augmentation of bone formation was noticed which was not the case in the control (untreated) group. The hydro alcoholic extracts are more active than the aqueous extracts. The aqueous and hydro alcoholic extracts are nontoxic. Advantages of the invention: The invention provides a safe and convenient product, which can be taken for healing the bone injuries. The invention provides that the drug is made available throughout the country and at all time. The availability does not depend on the location or season of the year. The method of extraction prevents any toxic materials getting extracted and present in the final product. This facilitates avoiding the intake of the toxic materials thereby preventing any harmful effects on the patient. We claim 1. A Novel extract from the herb Asvstasia gangetica (family: Acanthaceae) having properties of healing bone injuries comprises I. Salts of sodium in the range of 0.4 to 2.4 % by weight. II. Salts of potassium in the range of 3.5 to 6.1% by weight. III. Salts of calcium (except calcium oxalate) in the range of 1.0 to 5.5% by weight. IV. Salts of magnesium in the range of 0.12 to 2.5% by weight. V. Salts of zinc in the range of 0.001 to 0.004% by weight. VI. Sugars in the range of 8.4 to 29.3% by weight. VII. Polyphenols (flavonoids) in the range of 8.1 to 12.9% by weight and VIII. Other unidentified nontoxic agents in the range of 78.48 to 41.3% by weight 2. An extract as claimed in claim 1, wherein salts are preferably in the range of: sodium range from 0.6 to 2.1%, potassium range from 3.7 to 5.8%, calcium (other than calcium oxalate) range from 1.4 to 5.1%, magnesium range from 0.15 to 2.1% zinc range from 0.002 to 0.003%. 3. An extract as claimed in claims 1 to 2, wherein the amount of sugars, preferably in the range from 10 to 25%. 4. An extract as claimed in claims 1 to 3, wherein the amount of polphenols, ranges from 9.0 to 11.5%. 5. An extract as claimed in claims 1 to 4, wherein other unidentified nontoxic agents range from 75 to 48.4 %. 6. An extract as claimed in claims 1 to 5, wherein the salts are present as chlorides, sulphates, lactates, tartarates, citrates and the like. 7. A process for the preparation of an extract from the herb Asvstasia gangetica (family: Acanthaceae) having properties of healing bone injuries as defined above which comprises a) Powdering the parts of the herb Asvstasia gangetica (family: Acanthaceae) b) Extracting the resulting powder with a polar or a mixture of polar solvents such as distilled water, alcohol or aqueous alcohol in the range 5 to 20% at a temperature in the range of 35-45°C using a water bath c) Maintaining the temperature of tiie bath strictly at the above said temperature while stirring continuously for a period in the range of 2 to 6 hrs. d) Filtering the resulting extract to remove any suspended materials e) Washing the residue with the same solvent or a mixture of solvents employed in the step b above f) Concentrating the filtrate by Heating the filtrate in the water bath by maintaining the temperature in the range of 45-55°C g) And if desired, forming pills / tablets / capsules / liquid orals / by conventional methods. 8. A process as claimed in claim 7 wherein the solvent used for extraction is selected from dematerialized water or hydro alcoholic mixture. 9. A process as claimed in claims 7 & 8 wherein the parts of the herb used for extraction is selected from leave or the stem or the mixtures and the sizes range from 20 to 40- mesh size. 10. An extract form the herb Asvstasia gangetica (family: Acanthaceae) having properties of healing bone injuries substantially as herein described with particular reference to the examples. 11 .A process for the extraction of an extract from the herb Asvstasia gangetica (family: Acanthaceae) having properties of healing bone injuries substantially as herein described with particular reference to the examples.

Full Text

This invention relates to an extract from an herbal plant having bone fracture healing properties. The invention particularly relates to an extract from the herb Asvstasia gangetica (family: Acanthaceae). This invention also relates to a process for the preparation of the above said extract having bone fracture healing properties from the herb Asvstasia gangetica.
The prolonged period that is generally required for healing of fractures is a matter of great concern and frustration both to the patient and to the orthopedic surgeon. Treatment of bone fracture often involves surgery, which may prove hazardous to very elderly patients. Even after treatment one can't be certain whether a particular fracture gets healed normally or results in delayed union or even non-union. Because of these uncertainties the patient may have to wait for a prolonged period before he can use the fractured limb properly. Besides applying plaster with or without surgery, there is no rational clinical treatment to hasten the healing process although vitamins and anabolic hormones can play a useful role especially when the patient is deficient in them.
The main objective of the present invention is therefore to provide an extract from the herb Asvstasia gangetica having bone fracture healing properties. Another objective of the present invention is to provide a process for the preparation of an extract from the leaves /stems of the herb Asvstasia gangetica having bone fracture healing properties.
The present invention provides a process for the preparation of a medicament from the leaves and stems of Asvstasia gangetica (family: Acanthaceae) by extraction with water or aqueous alcohol at controlled temperature.
So far only one herb namely Cissus guadrangularis (family: Vitaceae) has been reported [K.N.Udupa, H.J.Arnikar, L.M.Singh - Experimental studies of the use of Cissus

auandrangularis in healing fracture, Indian.J.Med.Sci, 1961,l5,551;K.N.Udupa and G.C.Prasad - Further Studies on the effect of Cissus quadrangularis in accelerated fracture healing, Indian J.Med.Res, 1964,52,1] to have bone fracture healing property. Chemical analysis of this herb revealed a high concentration of vitamin C besides carotene, calcium oxalate and a substance like "anabolic steroid'. It may be mentioned that calcium oxalate is an undesirable substance in many herbs, at times in high concentration. Excessive ingestion of calcium oxalate will lead to the complaint oxaluria. While vitamin C and carotene have nutritive effect, the nature and effect of the "anabolic steroid" is not known till now. Since ancient times the herb has been in use, especially by bone setters in villages for both external application as paste and as an internal medicine to be taken with milk. For scientific investigation an alcoholic extract of the herb was obtained by cold percolation method, drying it and then dissolving it in ethylene glycol and administering by intramuscular injection. This has hastened fracture healing by reducing the total convalescent period by 33% in the experimental animals.
In subsequent animal experiments [K.N.Udupa and G.C. Prasad, Biomedical and Calcium'*^ studies on the effect of Cissus quadrangularis in fracture repair, Indian J.Med.Res, 1964, 52, 480; K.N.Udupa. G.C.Prasad and S.P.Sen, The effect of phytogenic anabolic steroid in the acceleration of fracture repair. Life science, 1965, 4, 317]. Both the aqueous alcoholic extract and a lipid soluble "phytogenic steroid" (anabolic steroid) were found to possess bone fracture healing property when given by intra peritoneal route. Both have shown toxicity when the dosage was increased.
Till now neither the aqueous alcoholic extract nor the "phytogenic steroid" have been developed for clinical studies and therapeutic use. Except this herb no other herb has been investigated nor generally known to have bone fracture healing properties.
Asystasia gangetica (Linn) (synonym: Asvstasia coromandeliana) family: Acanthaceae) is an erect unarmed glabrous herb, 50 to 100 cm.high, branches arising

from the base, leaves orate, entire, glabrous, subacuminate 4 x 1.5 - 2.5 cm. Kacemes up to 30 cm. long; corolla white capsules enclosed within the persistent calyx teeth. It is an abundant weed during the rainy season forming large patches in open places. In Sanskrit it is called LAVANAVALLI. In medicinal plant literature [R.N.Chopra, S.L.Nayar and I.e. Chopra, Glossary of Indian Medicinal Plants, Publications and Information Directorate, C.S.I.R., New Delhi, p.30 (1956)]. It was mentioned that the juice of the plant is anthelmentic, given in rheumatism and swellings.
The use of the leaves of the above mentioned herb for healing of fracture is practiced in Haryana. As per this practice the personnel who are beaten by police with lathis is asked to take the dried powder of the leaves of the said herb along with milk. It is the belief of the people in that State that by doing so the personnel inflicted with injuries to the bone as a result of such beatings get relief. Such a practice is not prevalent in other parts of the country. There is no published information about this practice based on the belief followed in Haryana.
Another peculiarity of this herb is that it is not available throughout the year and throughout India, The plant grows only in rainy season and that too only in certain parts of India like Haryana and Andhra Pradesh. This means the herb is location and climate specific. Keeping in mind the folklore use of the leaves of the herb, and the importance of providing relief for the patients suffering from bone injuries, we took the initiative to study the medicinal properties of this herb in a systematic way.
Our detailed studies on the herb revealed that in addition to the presence of active ingredients, responsible for imparting relief to the injuries inflicted to the bones, in the parts of the herb, especially the leaves, it also contains insoluble salts of calcium like calcium oxalate and toxic metals like lead, arsenic and mercury which are toxic and therefore injurious to health. Therefore by continuous intake of the raw leaves directly, though there is immediate relief of the injuries to the bones, due to the presence of above said toxic materials, which also simultaneously adversely affects the health of the patient in a longer run leading to other complications. These dangerous consequences are not

understood or realized by those who are traditionally practicing the use of the said herb for healing bone injuries.
Our systematic and scientific studies on the bone healing properties of the herb Asvstasia gangetica such as leaves, stems, flowers, roots, etc. Indicates that the parts of the herb are rich in minerals and organic constituents, which are responsible for imparting such properties to the herb. The main constituents which imparts the bone healing properties to the parts of the herb, particularly the leaves and the stems are found to be the salts of sodium, potassium, calcium (other than calcium oxalate), magnesium and zinc and organic constituents like sugars and polyphenols (flavonoids). These active ingredients are present in specific amounts.
Hence the inventive step lies in identifying the main constituents for bone healing properties from the Asvstasia gangetica. Accordingly The main constituents which imparts the bone healing properties to the parts of the herb, particularly the leaves and the stems are found to be the salts of sodium, potassium, calcium (other than calcium oxalate), magnesium and zinc and organic constituents like sugars and polyphenols (flavonoids). These active ingredients are present in specific amounts.
Therefore the main objective of the present invention is to provide an extract having
bone fracture healing properties from the herb Asvstasia gangetica (family:
Acanthaceae)
Another objective of the present invention is to provide a process for the preparation of the extract having bone fracture healing properties from the herb Asvstasia gangetica (family: Acanthaceae)
The principle by which the invention has been developed is based on the folklore use of the herb Asvstasia gangetica (familv: Acanthaceae) practiced in Haryana and by extracting the parts of the said herb employing a specific method which will extract only

the organic and inorganic active ingredients wliicli do not undergo any deconiposition and avoiding extraction of any of the toxic ingredients present therein.
Accordingly the present invention provides an extract from the parts of the herb, Asystasia gangetica (family: Acanthaceae) having bone fracture healing properties comprising
I. Salts of sodium in the range of 0.40 to 2.4% by weight.
II. Salts of potassium in the range of 3.5 to 6.1%. By weight.
III. Salts of calcium (excluding calcium oxalate) in the range of 1.0 to 5.5%. By weight.
IV. Salts of magnesium in the range of 0.12 to 2.5%. By weight.
V. Salts of zinc in the range of 0.001 to 0.004% by weight.
VI. Sugars in the range of 8,4 to 29.3%. By weight.
VII. Polyphenols (flavonoids) in the range of 8.1 to 12.9 %. By weight.
Vlil. Other unidentified nontoxic agents in the range of 78.48 to 41.3%. By weight
The metals mentioned above are present in the form of their salts (other than oxalate) like chloride, sulphate, lactate, tartarate and citrate. They are estimated and expressed as metals throughout.
The amount of the above mentioned active constituents vary depending upon the parts used for extraction. The salts of the metals may be present as chloride, sulphate, lactate, citrate, tartrate, etc. It is important to note that the extract does not contain calcium oxalate and other toxic metal salts of Arsenic, Lead, Mercury, and Cadmium etc., which are usually present in the parts of the herb. The sugars and flavonoids have not yet been identified. There is likely that some of these agents may be novel. Similarly work is in progress to identify the other unidentified non-toxic components. It may be possible that all or some of these agents may turn out to be new agents.
The amount of the salt of sodium may preferably range from 0.6 to 2.1% by weight. The amount of the salt of potassium may preferably range from 3.7 to 5.8% by weight.

The amount of the salt of calcium (other than toxic calcium salts) may preferably range
from 1.4 to 5.1% by weight.
The amount of the salt of magnesium may preferably range from 0.15 to 2.1% by weight.
The amount of the salt of zinc may preferably range from 0.002 to 0.003% by weight.
The amount of sugars may preferably range from 10 to 25% by weight.
The amount of polyphenols (flavonoids) may preferably range from 9 to 11.5% by weight.
According to another feature of the present invention there is provided a process for the preparation of the above-mentioned extract from the parts of the herb, Asvstasia qangetica (family Acanthaceae) having bone fracture healing properties comprising.
I. Powdering the dried parts of the herb Asvstasia gangetica (family :Acanthaceae)
II. Extracting the resulting powder with a polar solvent or a mixture of polar solvents at a temperature in the range of 35 to 45 0C using a water bath.
III. Maintaining the temperature of the bath strictly at the above said temperature while stirring continuously for a period in the range of 2 to 6 hrs
IV. Filtering the resulting extract to remove any suspended material
V. Washing the residue with the same solvent or mixture of solvents used for the extraction.
VI. Concentrating the resulting filtrate under reduced pressure by heating in a water bath maintaining the temperature in the range of 45 to 55°C and if desired.
VII. Forming pills /tablets /capsules/liquid orals of the resulting product by conventional methods.
In a preferred embodiment of the invention the polar solvent employed for extraction may be selected from distilled water, dematerialized water, alcohol or aqueous alcohol wherein the ratio of water to alcohol ranges from 5 to 20%.
The sizes of the powder of the parts of the herb may range from 20 to 40 meshes.
The resulting product which is in the form of soft or dry extract (containing 10-20% water) may be preserved by adding a preservative and can be made into

Pills/tablets /capsules/liquid orals by the conventional meinoas.
The invention is disclosed in detail in the examples given below which are provided by way of illustration only and therefore should not be construed to limit the scope of the invention.
Example-1.
The fresh leaves collected from Asvstasia gangetica herb are shade dried and powdered coarsely (20-40 mesh) in an electric grinder. The coarse powder (500 g) is taken into a 5 I. beaker and 3000 ml. dematerialized water is added. An electric paddle stirrer is introduced into the beaker. The beaker is kept in an electric water bath with thermostatic control. The water bath temperature is maintained in the range of 35-40'C with the help of thermostat. The extraction is carried out under continuous stirring for 4 hours. There after the solvent is filtered under suction and washed with 250 ml dematerialized water.
The filtrate is once again filtered through a funnel containing a tight cotton plug to remove all suspended material. The clear filtrate is concentrated in a rotary evaporator over a water bath maintained at 45-50°C. The resulting product is a gummy solid soluble in water, yield 5.5%.
The product contains salts of sodium 0.7%, potassium 3.5%, calcium (except calcium oxalate) 2.1%, magnesium 1.5 %, and zinc 0.002%, and sugars (reducing substances calculated as glucose) 8.4% and polyphenols (flavonoids) 8,1%.
Pills for oral administration may be prepared after thoroughly mixing the extract with known pharmaceutical solids and /or liquid excipients and then rolling the mass into a rod of even thickness, cutting to the length required and rounding to obtain perfect spheres.

Example-2
The stems of Asvstasia ganqetica are separated from leaves and shade dried. The dried stems are broken into small pieces and powdered in an electric grinder to a coarse powder (20-40 mesh). The coarse powder (450 g) is taken into a 5 I beaker and 2500 ml of distilled water is added. An electric paddle stirrer is introduced into the beaker. The beaker is kept in thermostat water bath maintained at 40-45°C. The extraction is carried out under continuous stirring for 3 hours.
There after the solvent is filtered under reduced pressure and washed with 300 ml of distilled water. The filtrate is once again filtered through a large funnel using a fluted filter paper. The clear filtrate in concentrated in a film evaporator at 40"45°C to remove all water. The resulting product is a gummy solid soluble in water, yield 5.7%.
The product contains salts of sodium 2.1%, potassium 5.3%, calcium (except calcium oxalate) 5.3%, magnesium 1.3 %, and zinc 0.003%, and sugars (reducing substances calculated as glucose) 9% and polyphenols (fiavonoids) 9.6%.
Tablets for oral administration may be prepared from the extract by direct compression or wet granulation after dilution with microcrystalline cellulose, lactose, starch and other such materials used as diluents
Example-3
The coarsely powdered dried leaf of Asvstasia gangetica (400 g) is treated with 10% aqueous alcohol (3500ml) (alcohol to water ratio 1:9 by volume) in a 5 I beaker. The beaker is immersed in an electric water bath having a thermostat maintained at 35-40°C. An electric stirrer is introduced into the beaker and the extraction is carried out with continuous stirring for 5 hours. Periodically small volumes (20-30 ml) of 10% alcohol are added to make good the loss of solvent by evaporation during extraction. Thereafter the solvent was filtered and the residue washed with 300 ml 10% aqueous alcohol. The clear

filtrate was concentrated under reduced pressure at a temperature of 45-50°C to remove all alcohol. The aqueous concentrate is then freeze dried when the product was obtained as hygroscopic form which is freely soluble in water, yield 7.2%.
The product contains salts of sodium 0.6%, potassium 5.9%, calcium (except calcium oxalate) 2.2%, magnesium 2.3 %, and zinc 0.002%, and sugars (reducing substances calculated as glucose) 13.4% and polyphenols (flavonoids) 11.5%.
Soft gel capsules for oral administration may be prepared from the extract by dilution with suitable hydrophobic diluents such as corn oil or other vegetable oils and filled by known methods using rotary die process. Hard gelatin capsules may be prepared after adding moisture absorbing diluents like starch, anhydrous declaim phosphate and the like and filling the capsules by known methods.
Example-4
The coarsely powdered dried stem of Asvstasia qanqetica (500 g) is treated with 15 % aqueous alcohol (3200 ml) (alcohol to water ratio 3: 17 by volume) in a two-necked 5 I round bottom flask fitted with a stirrer and a condenser. The round bottom flask is immersed in an electric water bath with a thermostat and the temperature maintained at 38-45°C. Extraction is carried out under continuous stirring for six hours. Thereafter the solvent was filtered through a thin bed of washed kieselguhr through a large Buchner funnel and the residue washed with 400 ml of 15 % aqueous alcohol. The clear filtrate was concentrated at 45-50°C in rotary evaporator to a dry extract containing ca.10% moisture. The resulting product is a gummy material freely soluble in water, yield 7.5%.
The product contains salts of sodium 0.6%, potassium 5.3%, calcium (expect calcium oxalate) 1.9%, magnesium 1.0 %, and zinc 0.001%, and sugars (reducing substances calculated as glucose) 27.2% and polyphenols (flavonoids) 11.8 %.

Oral liquid dosage form may be prepared by dissolving or dispersing ine extract in a suitable non-medicated syrup. The non-medicated syrup contains sweetening agent flavors, optionally containing colorants and ant microbial preservatives.
Test results of the bone healing properties of the product obtained from examples 1 to 4 of the invention:
The aqueous extracts obtained by examples 1 and 2 or hydroalcoholic extracts obtained by examples 3 and 4 when administered orally as aqueous solution/suspension to rats in which fracture was produced artificially in the right tibia and fibula showed enhanced bone healing properties compared to control group after one week. After two weeks augmentation of bone formation was noticed which was not the case in the control (untreated) group. The hydro alcoholic extracts are more active than the aqueous extracts. The aqueous and hydro alcoholic extracts are nontoxic.
Advantages of the invention:
The invention provides a safe and convenient product, which can be taken for healing the bone injuries. The invention provides that the drug is made available throughout the country and at all time. The availability does not depend on the location or season of the year. The method of extraction prevents any toxic materials getting extracted and present in the final product. This facilitates avoiding the intake of the toxic materials thereby preventing any harmful effects on the patient.

We claim
1. A Novel extract from the herb Asvstasia gangetica (family: Acanthaceae) having
properties of healing bone injuries comprises
I. Salts of sodium in the range of 0.4 to 2.4 % by weight.
II. Salts of potassium in the range of 3.5 to 6.1% by weight.
III. Salts of calcium (except calcium oxalate) in the range of 1.0 to 5.5% by weight.
IV. Salts of magnesium in the range of 0.12 to 2.5% by weight.
V. Salts of zinc in the range of 0.001 to 0.004% by weight.
VI. Sugars in the range of 8.4 to 29.3% by weight.
VII. Polyphenols (flavonoids) in the range of 8.1 to 12.9% by weight and
VIII. Other unidentified nontoxic agents in the range of 78.48 to 41.3% by weight

2. An extract as claimed in claim 1, wherein salts are preferably in the range of: sodium range from 0.6 to 2.1%, potassium range from 3.7 to 5.8%, calcium (other than calcium oxalate) range from 1.4 to 5.1%, magnesium range from 0.15 to 2.1% zinc range from 0.002 to 0.003%.
3. An extract as claimed in claims 1 to 2, wherein the amount of sugars, preferably in the range from 10 to 25%.
4. An extract as claimed in claims 1 to 3, wherein the amount of polphenols, ranges from 9.0 to 11.5%.
5. An extract as claimed in claims 1 to 4, wherein other unidentified nontoxic agents range from 75 to 48.4 %.
6. An extract as claimed in claims 1 to 5, wherein the salts are present as chlorides, sulphates, lactates, tartarates, citrates and the like.
7. A process for the preparation of an extract from the herb Asvstasia gangetica (family: Acanthaceae) having properties of healing bone injuries as defined above which comprises

a) Powdering the parts of the herb Asvstasia gangetica (family: Acanthaceae)
b) Extracting the resulting powder with a polar or a mixture of polar solvents such as distilled water, alcohol or aqueous alcohol in the range 5 to 20% at a temperature in the range of 35-45°C using a water bath

c) Maintaining the temperature of tiie bath strictly at the above said temperature while
stirring continuously for a period in the range of 2 to 6 hrs.
d) Filtering the resulting extract to remove any suspended materials
e) Washing the residue with the same solvent or a mixture of solvents employed in
the step b above
f) Concentrating the filtrate by Heating the filtrate in the water bath by maintaining the
temperature in the range of 45-55°C
g) And if desired, forming pills / tablets / capsules / liquid orals / by conventional
methods.
8. A process as claimed in claim 7 wherein the solvent used for extraction is selected
from dematerialized water or hydro alcoholic mixture.
9. A process as claimed in claims 7 & 8 wherein the parts of the herb used for extraction
is selected from leave or the stem or the mixtures and the sizes range from 20 to 40-
mesh size.
10. An extract form the herb Asvstasia gangetica (family: Acanthaceae) having properties
of healing bone injuries substantially as herein described with particular reference to
the examples.
11 .A process for the extraction of an extract from the herb Asvstasia gangetica (family: Acanthaceae) having properties of healing bone injuries substantially as herein described with particular reference to the examples.

Documents:

2121-mas-1998-abstract.pdf

2121-mas-1998-claims filed.pdf

2121-mas-1998-claims granted.pdf

2121-mas-1998-correspondnece-others.pdf

2121-mas-1998-correspondnece-po.pdf

2121-mas-1998-description(complete) filed.pdf

2121-mas-1998-description(complete) granted.pdf

2121-mas-1998-form 1.pdf

2121-mas-1998-other documents.pdf

« Previous Patent

Next Patent »

Patent Number

209018

Indian Patent Application Number

2121/MAS/1998

PG Journal Number

38/2007

Publication Date

21-Sep-2007

Grant Date

16-Aug-2007

Date of Filing

22-Sep-1998

Name of Patentee

M/S. NATCO PHARMA LTD

Applicant Address

ROAD NO 2,BANJARA HILLS, HYDERABAD-500 033.

Inventors:

#	Inventor's Name	Inventor's Address
1	VENKATA RAO ERRAGUNTLA	ROAD NO 2,BANJARA HILLS, HYDERABAD-500 033.

PCT International Classification Number

A 61 K 35/78

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA