Title of Invention	"A SYNERGISTIC HERBAL COMPOSITION"
Abstract	The present invention generally relates to a therapeutic synergistic herbal composition for Neutropenia and a method for the treatment of Neutropenia using the herbal composition.

Full Text

Field of the Invention The present invention relates to a synergistic herbal composition comprising Withania somnifera and Shilajit. The present invention generally relates to a therapeutic synergistic herbal composition for Neutropenia and a method for the treatment of Neutropenia using the herbal composition. Background of the Invention One of the popular measures of controlling cancer is administration of chemotherapy. Chemotherapy works by acting on the quickly dividing cancerous cells. However, chemotherapy cannot distinguish between tumor and normal cells, such as bone marrow cells, neutrophils, hair follicles, etc. which also divide quickly. Bone marrow cells are important as they produce WBCs, RBCs and neutrophils and other cells. Decrease in bone marrow cells or myclosuppression is therefore an inevitable consequence of chemotherapy. A patient who takes chemotherapy invariably suffers from a condition called neutropenia, wherein the blood neutrophil count decreases. White blood cells and neutrophils get killed during chemotherapy, causing significant reduction in their count and also consequently weakening the immune system. A fall in WBC count leads to an increased risk of infections. Neutropenia is an important problem associated with chemotherapy. Majority of cytotoxic drugs also cause suppression of the bone marrow cells. The bone marrow has a storage compartment that can supply mature cells to the peripheral blood for 8 to 10 days after the stem cell pool has been damaged by drugs. Events measured in the peripheral blood are usually a week behind those occurring in the bone marrow. Since chemotherapy results in decrease in blood neutrophil count, it is a basic accepted tenet that the degree of neutropenia and the degree of chemotherapy go hand in hand. In untreated patients, leukopenia and thrombocytopenia are discernible on the ninth or tenth day after initial dosing. Nadir blood counts are noted between days 12 to 18 with recovery apparent by day 21 and complete recovery being observed by day 28, with notable exceptions. For most regimens of chemctherapy, the adjustment of dose is defined and often both, blood counts at the nadir and at the start of the next course of treatment are taken into account. When an unexpectedly low nadir is found, the dose of the bone-marrow-suppressing drugs is decreased. A low count before the next course of treatment can be a reason to either lower the dose or postpone the treatment. For example, dose reductions of 20% which are not infrequent in clinical oncology practice are usually related to myelosuppression and can lead to a 50% reduction in cure rate. Prior treatment with drugs or radiation may alter this sequence by depleting the stem cell pool, shortening the time to the appearance of leukopenia and thrombocytopenia and prolonging the recovery time. The interval of greatest importance in the clinic is the duration of the nadir level of leukocytes. The highest risk of infection occurs with granulocyte counts lower than 500/mm3. If this nadir lasts only 4 to 7 days, it is tolerated by most patients without supplemental support. Increasing the doses of most anticancer drugs within the range of the maximally tolerated dose usually does not ablate the marrow or even prolong the recovery time. Repeated dosing during the phase of early recovery of the marrow (days 16-21) however, may cause severe toxicity in the second treatment cycle in patients whose marrow is not the source of or involved with the tumor. Several workers have conducted studies to overcome the problem of neutropenia encountered during chemotherapy. One attempt to overcome this problem is by introducing the concept of 'prohost therapy'. This approach aims at administering drugs to bolster immune defenses against infections. Several naturally occurring and synthetic substances like BCG, Corynebacterium parvum, mauramyl dipeptide, glucans, interleukin-1 and colony stimulating factors have been used to enhance the nonspecific host resistance against infections. Various hormones that regulate hematopoiesis are known as colony stimulating factors (CSF) or hematopoietins. The primary reason for using growth factors is to improve or optimize the patient's host defense capabilities. Although most clinicians are comfortable with the concept of replacement therapy, such an approach does not play a major role in hematopoietic growth factor treatment in cancer. Using hematopoietic growth factors it is possible to regulate the production of erythrocytes, neutrophils, eosinophils and monocytes. They either affect the proliferation of haemopoietic progenitor cells or have an indirect action producing their effects by induction of cytokines from accessory cells. Growth factors have been coupled with cytotoxic combination chemotherapy and the nadir leukopenia can usually be avoided or ameliorated. The colony stimulating factors are granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), and erythropoietin (EPO). These glycoproteins have been cloned through DNA technology and three of them (viz., G-CSF, GM-CSF, and EPO) have been approved by the FDA for clinical use. There are two main classes of CSFs. Class 1 CSFs such as IL-3 or GM-CSF act at the partially committed stem cell level to cause differentiation and proliferation of multiple cell lines (monocytes, granulocytes, eosinophils etc.). Class 2 CSFs such as G-CSF, M-CSF, and erythropoietin act on already differentiated cell lines to stimulate proliferation of more specific cell types. A survey of the literature reveals that there are very few approaches using herbal products to counter the problem of neutropenia, leukopenia and thrombocytopenia. Thus, there exists a need to develop herbal compositions having applications in this sphere of treatment. Accordingly, the Applicant, conducted a detailed investigation on several herbs and surprisingly found a novel herbal composition which can be used to treat neutropenia. The ingredients of this composition are Shilajit and an extract from the herb Withania somnifera. Shilajit is a black brown natural exudate obtained from rocks during hot weather in the lower Himalayas, Vindhya, Nepal and other mountain tracts. It is 'generally a compact massof vegetable organic matter, composed of gummy matrixinterspersed with vegetable fibres and minerals. Shilajit is purified in accordance with the process described in Ayurvedic literature before being formulated for human consumption. It contains nutritive minerals, certain essential amino acids and a range of other organic constituents in minute quantities. Other substances found in Shilajit include potassium, nitrogen, silica, iron, sulphur, iodine, etc. Literature does indicate that Shilajit exhibits immunomodulatory activity and it may have the ability to promote growth and proliferation of cells. Withania somnifera Dunal also known as Ashwagandha, is an erect shrub belonging to the Solanaceae family. Withania somnifera Dunal is commonly used in many indigenous preparations as an immunostimulant (Ghosal, S., Lai, J.,Srivastava, R., Bhatacharya, S.K., Upadhyay, S.N., Jaiswal, A.K. & Chattopadhyay, U (1989): Immunomodulatory and CNS effects of Sitoindosides IX & X, two new glycowithanolides from Withania somnifera. Phytother. Res. 3(5) pp. 201-206). The extract has been reported to increase the hemoglobin, total red blood cells and white blood cells in mice with chemically suppressed immunity. Moreover, the extract has also been found to stimulate the total WBC and bone marrow cells in animals treated with a non-lethal dose of radiation (Kuttan, G. (1996). Use of Withania somnifera Dunal as an adjuvant during radiation therapy, Indian J. Exp Biol Sep: 34(9): 854-6. Devi, P.U. (1996). Department of Radiobiology, Kasturba Medical College, Manipal, India, Withania somnifera Dunal (Ashwagandha): potential plant source of a promising drug for cancer chemotherapy and radiosensitization, Indian J. Exp Biol Oct.34 (10): 927-32.). An inverse correlation exists between the level of circulating neutrophils and the frequency of infections. In addition to the absolute decrease in the number of neutrophils, a decrease in mobilization into areas of inflammation occurs in severe neutropenic disorders. Pretreatment for 15 days with Withania somnifera extract produced a striking leucocytosis with a predominant neutrophilia. Withania somnifera extract has been found to protect mice against cyclophosphamide-induced neutropenia (Thatte U.M., Chhabria, S.S., Karandhikar, S.M., and Dahanukar, S.A. (1987b). Protective effects of Indian medical plants against cyclophosphamide neutropenia. J. Post. Med. 33, 185). The WBC count attained a normal value in (10 days) in comparison to cyclophosphamide alone group who did not regain a normal value even after 30 days (Davis, L., Kuttan, G. (1998). Suppressive effect of cyclophosphamide-induced toxicity by Withania somnifera extract in mice, J.Ethnopharmacol . Oct., 62(3) 209-14) . The exact mechanism of action is not clear yet. It produces neutrophilia and leucocytosis. Thus, although the prior art does indicate that Withania somnifera has some positive effect on the immune system, its potential for treatment of neutropenia or the effect of Shilajit on neutropenia has not been studied. Also, there is a pronounced need in the art for the development of a herbal composition for combating the side-effects of chemotherapy, especially neutropenia. Objects of the Invention The main object of the invention is to provide a novel synergistic herbal composition useful in the treatment of neutropenia. Another object of the invention is to provide a method for the production of the herbal composition. Yet another object of the invention is to provide a method for the treatment of Neutropenia using the novel herbal composition. Summary of the invention Accordingly, the invention provides a novel synergistic herbal composition comprising effective Shilajit and Withania somnifera for the treatment of neutropenia. The invention also provides a process for the production of the said composition and a method for the treatment of neutropenia. Detailed description of the Invention In accordance with the above and other objects, the invention provides a novel synergistic herbal composition comprising therapeutically effective amount of Shilajit and Withania somnifera, for the treatment of neutropenia. As discussed in the foregoing sections, the prior art indicates that Withania somnifera has some positive effect on the immune system. It is capable of promoting the growth of white blood cells (WBCs) in the bone marrow. However, the bone marrow is composed of several distinct cells apart from red blood corpuscles (RBCs).and WBCs, such as myeloblasts, immature stem cells etc. The potential of Withania somnifera to promote the growth and proliferation of all these cells is being investigated. Further, the potential of Shilajit to promote the growth and proliferation of all these cells is also not hitherto known. The Applicant conducted a detailed study on the effect of Shilajit and Withania somnifera on the immune system, especially during the period the patient is undergoing chemotherapy. On an experimental basis, a tablet containing only Shilajit and only Withania somnifera were administered to separate groups of subjects undergoing chemotherapy. Yet another group was given tablets containing Shilajit and Withania somnifera. The doses of administration of these tablets varied from 100 mg to 400 mg. The Applicant found that subjects who consumed tablets containing only Shilaji or Withania somnifera showed initial rise in the blood count at a particular dose and the count fell thereafter. The observation was as expected since chemotherapy has an adverse effect on the immune system. However, to the utter shock and amazement of the Applicant, the subjects who consumed tablets containing Shilaji and Withania somnifera showed an initial rise in the blood count, and this rise was maintained throughout the period of treatment, as long as chemitherapy was being used. This observation was totally contrary to the basic tenet that decrease in neutrophil count occurs simultaneous with administration of chemotherapy. This finding is described in detail in the experiments that follow in the 'Example' section. It was this amazing and surprising finding that led the Applicant to conclude that a herbal composition containing Shilajit and Withania somnifera can be used to treat neutropenia. From the above observation, the Applicant also concluded that the ingredients of the composition exhibit a synergistic effect, in the sense that it is only a composition containing Shilaji and Withania somnifera that is capable of maintaining and increasing the neutrophil count in the blood even with concurrent administration of chemotherapy. The ingredients of the composition exhibit a far greater therapeutic effect on the bone marrow cells than the individual ingredients thereof. The composition and the synergistic effect thereof is described in detail in the Examples. One of the ingredients employed in the herbal composition of the invention is Withania somnifera, specifically, the extract of Withania somnifera. The extract may be obtained from the leaves, roots, fruits, flowers or bark of Withania somnifera. The powder of the parts of this plant may also be used in the herbal composition instead of the extract but its dosage will be too high.. For advantageous and best results, the hydroalcoholic extract from the roots of the plant Withania somnifera may be used. Typically, the roots of Withania somnifera are dried and ground to a powder, which is then extracted with hot water, although aqueous alcoholic solutions may also be used. Prior to use in the herbal composition of the invention, Shilajit is to be processed. The rock exudate (raw Shilajit) is purified first with water, by boiling for approximately 2-3 hours. The process of boiling is repeated 2-3 times by which time, the impurities get removed. The decoction so obtained is allowed to settle for 72 hours. This decoction of Shilajit is thereafter, optionally purified by boiling with Triphala (Emblica officinalis, Terminalia chebula and Terminalia belerica), cooling and concentrating the extract to obtain a powder of Shilajit, which can be used for administration. The synergistic herbal composition of the invention contains a therapeutically effective amount of extract of Withania somnifera and Shilajit. "Therapeutically effective amount" in this composition describes an amount or concentration of the ingredient of the composition, which amount is capable of stimulating proliferation of bone marrow cells, especially neutrophils and myeloblasts. These effective amounts of the ingredients can be readily determined by a person skilled in the art, although, as a preferred embodiment, approximately equal portions of extract of Withania somnifera and Shilajit are used in the herbal composition of the invention. These ingredients may also be taken in proportions such as 3:2, 1:4, etc, although 1:1 has been found advantageous. As said earlier, either the extract or powder of the plant parts of Withania somnifera may be used in the composition. The composition is typically an intimate combination of the ingredients. The composition may be formulated into various physical forms suitable for oral administration such as tablets, capsules or syrups, by methods known in the art. In addition to the above herbs, various pharmaceutically acceptable additives, excipients, antioxidants, carriers, preservatives and fillers may be present. Administration of the composition is oral. The use of the composition is recommended five days prior to commencement of chemotherapy. The therapy is to be continued during the administration of chemotherapy and also post-cessation of chemotherapy until the initiation of the next chemotherapy cycle. Depending upon the diet of the subject and other factors, and the level of neutrophils in the blood, the use of the herbal composition of the invention may be discontinued or maintained at lower dosage or resumed. Statement of the invention The present invention relates to a synergistic herbal composition comprising Withania somnifera and Shilajit useful for stimulating bone marrow cell proliferation activity wherein the ratio of Withania somnifera and Shilajit is 1:4 to 3:2. Brief description of the accompanying drawings Figure 1 (a) to (b) are graphical representations of ANC data of two patients upon administration of DRF230798A Figure 2 (a) to (c) are graphical representations of ANC data of two patients upon administration of DRF230798C Accordingly, this invention relating to the synergistic herbal composition and a method for the treatment of neutropenia is illustrated herebelow by the following examples and studies. Although the description above contains many specificities, these should not be construed as limiting the scope of the invention but as merely providing an illustration of the presently preferred embodiment of the invention. Thus the scope of this invention should be determined by the appended claims and their equivalents. EXAMPLE 1: Preparation of the herbal composition This Example describes the preparation of the herbal composition. The processes described below can be scaled up to produce larger quantities of extracts. The details provided for preparation of the following abstracts reflect the presently preferred method for extract preparations and should not be considered as limiting. 1. Shilajit: Shilajit raw is taken with six time water in a boiling pan and is boiled for three hour. The decoction is collected and the process is again repeated. The combined decoction is allowed to settle for 72 hour. This is purified with trifla herbs [that is amla (emblica officinalis), bahera (Terminalia chebula), and haritaki (Terminalia belerica)], are taken with water in a boiling pan. This is boiled for two hour and decoction is collected. The process is repeated again and two decoctions are mixed. Clear supernatant liquid obtained after settling of the Shilajit extract is mixed with trifla herbal decoction. The mixed decoction is concentrated and dried to get the solid powder. 2. Aswagandha (Withania somnifera Dunal) is prepared as follows: Hydro-alcoholic solution in the ratio of 20:80 (water: alcohol) is prepared separately. A weighed quantity of the coarsely powdered Aswagandha (Withania somniferd) herb is mixed with 8 times hydro-alcoholic solution and is kept under agitation for 4 hours with mild heating (about 40-45°C). The solution is then filtered. The filtrate is kept separately. The residue is again extracted with 6 times of hydro-alcoholic solution for about 4 hours. This is also filtered and both the filtrates are mixed together. This extract is then concentrated and dried to obtain a solid powder. The above powders are combined in approximately equal proportion and sifted through a suitable sieve (# 40) to get a fine powder, which represents the herbal composition of the invention. Example 2: Preparation of oral formulations Tablet The composition of example 1 is mixed with the required excipients like starch, microcrystalline cellulose and are passed through # 40 sieve. The sifted material is granulated with binder solution like starch paste. The granulated mass so obtained is dried in a suitable drier like electric tray drier or Fluid Bed Drier. The dried granules are passed through a sieve to get granules of desired particle size. Retention left over the sieve is further passed through a suitable mill like multimill or cadmill to obtain granules of desired particle size. Sifted granules are mixed with suitable lubricants like talc and magnesium stearate. The lubricated granules are compressed to form tablets in a suitable rotary tablet compression machine. Syrup The composition of example 1 is converted into syrup form as follows: Distilled mineral water is taken in a steam jacketed pan fitted with a stirrer. To this sugar is added while stirrer is on and dissolved with the aid of heat. Citric acid, preservatives like methyl paraben sodium and propyl paraben sodium are added to the above. By heating the mixture sugar get inverted and invert syrup is formed. The composition of example 1 is added to the above and is mixed with the help of a stirrer for some time. Suitable colours are dissolved in small quantity of water and added to above. The whole mass is mixed for some more time and filtered through a suitable filter to get the clear syrup. Capsules The composition of example 1 is mixed with suitable excipients like starch, microcrystalline cellulose and are passed through # 40 sieve. The sifted material is mixed with lubricants like talc and magnesium stearate. The blend so obtained is filled into capsules using suitable capsule filling machine. Example 3: test of synergistic activity of the herbal composition The bone marrow consists of several types of dividing cell which are not stem cells, viz. Myeloblast, erythroblasts etc. and not evaluated in the colony forming assay. It is found that the herbal composition of the invention is more effective on lineage committed progenitors. This activity results in more mature cells in blood circulation. The present novel composition is effective in promoting growth and proliferation of neutrophils thereby alleviating the undesirable side effects of chemotherapy. The following experiment illustrates the ability of the samples to induce proliferative activity for haematopoietic progenitors in the serum of treated animals and studies proliferation induced in bone marrow cells in animals treated with the samples. Experimental design Swiss Albino mice were fed orally daily with the following three samples for ten days at the end of which the animals were killed by cervical dislocation. Sample A: This is a sample containing the extract of Ashwagandha only Sample S: This is a sample containing the extract of Shilajit only Sample C: This is a sample containing the composition of the invention Bone marrow as well as blood was collected and examined. The bone marrow was processed for flow cytometry and blood was allowed to clot and the serum was separated for testing stem cell proliferative activity by colony forming assay. Untreated controls were maintained and processed as the treated ones. A slurry of the three samples was prepared in sterile distilled water and the mice were fed orally, every day. The animals were divided into groups of five animals each. Each samples was tested initially at 100 - 200 and 300 mg/kg body weight on a group consisting of five animals each. A common untreated control group was maintained for every set of experiment. The treatment lasted ten days after which the animals were killed by cervical dislocation. Bone marrow and blood was collected in sterile manner. Serum was separated from the blood after allowing it to clot. The bone marrow was processed for flow cytometry and the serum tested for haematopoietic stem cell colony forming activity by standard methods. 50ul of serum each from the treated and control group was used for the assay. In the second experiment, the samples were treated at 300,400 and SOOmg/kg.body weight. Results Serum colony stimulating activity Myeloid-monocytoid colonies/2xlOOOOOMNC - all kinds of cells Sample lOOmg/kg 200mg/kg 300mg/kg "S" 25+1-2 102+/-8 475+/-12 "A" nil nil 168+/-160 "C" nil nil nil Percent cells in"S",G2+M of cell cycle - The increase in neutrophils is desired. Increased proliferation of the bone marrow cells Sample lOOmg/kg 200mg/kg 300mg/kg "S" 23.0 23.05 23.0 "A" 23.0 23.1 23.75 "C" 23.8 23.50 26.0 Control 23.0 23.0 23.0 Conclusions Sample "S", being a sample containing extract of 'Shilajit' only exhibited immunostimulatory activity. It showed increased serum colony stimulating activity as compared to controls at 300 mg/kg. Assays for higher doses viz.400 and 500 mg/kg,clearly showed that 300mg/kg has highest activity and higher doses decrease the activity. Sample"A", being a sample containing extract of Ashwagandha also showed variable response in terms of colony stimulating activity. Therefore, further set of experiments were done to verify this in three separate batches of samples of "A". These samples showed consistent elevation to the extent of 150 colonies per two hundred thousand MNC at 500mg/kg.body weight. This increase is lesser than that obtained with sample"S". Sample "C", surprisingly did not show any increase in the colony stimulating activity as compared to controls Bone marrow proliferation data by flow cytometry shows that Sample "C'alone has increased proliferation of the bone marrow cells as seen by increased "S"+G2+M fraction. The other two samples "A" and "S" do not have activity. Interpretation of results The results are rather interesting because the sample which has the highest serum colony stimulating activity is not the one where bone marrow proliferation has increased. This can be explained by the fact that bone marrow consists of several dividing cell types which are not stem cells, viz. myeloblasts, erythroblasts etc. and not evaluated in the colony forming assay. Therefore, the activity induced by "C" is more effective on lineage committed progenitors. This activity will result in faster increase in more mature cells in circulation. Whereas, sample "S" and to a lesser extent "A", stimulate the immune system which result in production of the entire range of cytokines including IL-3 and GM-CSF. These factors have direct effect on stem cell proliferation and will result in replenishment of stem cells killed by chemotherapy. Most of the anticancer drugs have a myeloprotective effect due to their anti-proliferative action. This myelo suppressive effect is usually seen in two stages that is Acute and Chronic. Acute myelo suppression is due to destruction of immature proliferative cells in the bone marrow and Chronic is noted when the progenitor cells are themselves destroyed. In the experiments conducted on mice, the colony formation of progenitor cells and bone marrow proliferation index suggested an increased rate of proliferation of immature cells. It was observed that the herbal composition has no effect on colony formation. However, it does generate a high proliferation of immature cells. With this, it was hypothesized that this composition is useful in prevention of acute myelosuppression due to increase cell mass of immature proliferating cells. This property was further highlighted in the animal experiments which was further carried out wherein the preventive potential for myelosuppression was observed when the mice were challenged with chemotherapy. The combination proved very effective in allaying the suppression of Blood cell count and also when given continuously for duration of chemotherapy and beyond. It showed sustained potential for doing so. This was further proven and highlighted in human clinical pilot studies, the results have been shown further. Example 4: Clinical Trial From the data on animal studies, it was concluded that the herbal composition consisting of Shilajit and Ashwagandha will significantly produce bone marrow proliferation resulting in increased number of neutrophils in circulation. To confirm this hypothesis, a clinical trial has been conducted on different patients. Some subjects are randomized to Control Group, some are randomized to DRF 230798A and some are in DRF 230798C group. DRF 230798A: this a sample in the form of tablet of 300 mg Ashwagandha extract. DRF 230798C: This is a tablet prepared according to the invention and contains 150 mg each of Aswagandha and Shilajit extracts respectively. In order to see the myeloprotective response of these two formulations Absolute Neutrophil Counts (ANC) levels were monitored in all patients on Day 1,9 12 & Day 18. Most of the subjects in this trial have received Chemotherapy as Combination of Adriamycine + Cyclophospamide, Some have received Paclitaxel + Carboplatin, Paclitaxel + Cisplatin, Cyclophospamide + Cisplatin and Few have received Gemcitabine + Paclitaxel + Cisplatin. Group wise Description of the ANC level of subject is as; Control Group Cycle-1: ANC data of all patients is showing Lowest ANC on Day - 12 and then slight increase in count on day 18. (Table Removed) Cycle - 2; Neutrophil count of majority patients was going down as compare to counts of # 1. Over all data shows neutropenia is prominent in majority of patients. (Table Removed) Cycle-3: Mean ANC and day wise ANC counts are showing decline trend in # 3 as compare to # 1 . (Table Removed) Cycle -4; No patient showed any evidence of ANC below 1500/cmm. (Table Removed) Cycle-5: Here in this cycle neutropenia was prominent in three patients, but ANC was not below 1500/cmm. Out of these a patient no II showed ANC 1500/cmm & 1620/cmm on Day - 9 & Day - 12, but on day-18 it was all right. This patient was doing very nice in last 4 cycles but here she suffered from low ANC. (Table Removed) Cycle - 6; Almost all patients in Control group have showed evidences of Low Neutrophil counts some where in the study. (Table Removed) Compiled Data of Control Group: (Table Removed) DRF-230798A GROUP All patients in this group have received DRF-230798A at a dose of 2 tablets per day. Cycle-1; In this group is self-control for individual patient. Means no test drug in this cycle for any patient. Base line ANC values of all patients were very good. In some cases ANC was below 1500/cmm particularly on Day-9 & Day -12. In case of one particular patient mean ANC was below 1500/cmm. (Table Removed) Cycle - 2; When ANC values of Day (-5) were compared with Day - 1 values, some cases shows marginal increase in neutrophil count. (Table Removed) Cycle - 3; There is no evidence of neutropenia on any cycle day with mean ANC levels above 2700/cmm. (Table Removed) Cycle - 4; There is not any problem of ANC. (Table Removed) Cycle - 5; All subjects are absolutely fine in terms of mean ANC (Table Removed) Cycle - 6; (Table Removed) Compiled Data of DRF 230798 A : (Table Removed) DRF - 230798C Group: Cycle-1; Here in this group also cycle 1 is working as a control for individual patient. When looked at baseline neutrophil counts, it was found that all patient's ANC was in normal range. Only one patient (Patient No IX) did not have baseline data of ANC and same patient had low ANC on Day 9 (1780/cmm) and Day 18 (902/cmm). It shows patient is having problem of delayed Neutropenia in the beginning itself. Similarly Patient No VIII reported ANC - 1200/cmm on Day 18 of #1. One more patient (Patient No: X) reported ANC - 960/cmm on day 9 only. In general data shows drastic decrease in ANC values from Baseline to Day-1, 9 &12 of #1. In some cases Day 18 ANC shows slight rise and in some it has gone further below. In all cases mean ANC was more than 3200/cmm except Patient No IX. (Table Removed) Cycle - 2 When looked at the data of Day 1 to Day 18. No patient had ANC below 1500/cmm. All cases had mean ANC above 2600/cmm in # 2. Table Removed) Cycle - 3,4.5 & 6; Patient No IX who had problem of Low ANC in #1, had no problem in #2, # 4 & # 5. But in Cycle 3 on Day - 18 her ANC was 1330/cmm and in Cycle 6 on Day 12 ANC was 1060/cmm. But her mean ANC in all cycles was more than 3000/cmm. This is an excellent example of myello-protective action of Drug. In short Patient had problem in #1 and then after when she started taking DRF - 230798C, her counts were absolutely fine all cycles except Day 12 & 18 of Cycle 6. A Patient No VII had problem of low ANC i.e. 1802/cmm on day 18 of #1, But after study drug was given No reported neutropenia till # 6. A Patient No VIII who had low neutrophil count (1200/cmm) On Day 18 of #1 & 1210 /cmm on Day (-5) of # 2. But after taking study drug no neutropenia reported in # 2. (Data available till # 2 only). (Table Removed) Compiled Data of DRF 230798 C; (Table Removed) Inference: 1. As is evident from the data in different groups, Absolute Neutrophil Count increase was more in patients on DRF 230798 C. The order of different groups in term of rise in ANC is as follows: DRF 230798 C > DRF 230798 A > Control Group. 2. There is not much variation in the ANC level of patients on DRF 230798 C. In case of Patient No IX ANC level was increased from 1341 (Cycle 1) to 3039 (cycle 6). WE CLAIM 1) A synergistic herbal composition comprising Withania somnifera and Shilajit useful for stimulating bone marrow cell proliferation activity wherein the ratio of Withania somnifera and Shilajit is 1:4 to 3:2. 2) A herbal composition as claimed in claim 1 comprising an extract of Withania somnifera. 3) A herbal composition as claimed in claim 2 wherein the extract of Withania somnifera is an aqueous extract or powder of the plant parts of Withania somnifera. 4) A herbal composition as claimed in claim 2 wherein the extract of Withania somnifera is obtained from the roots, stem, leaves , fruits or flowers of the plant. 5) A herbal composition as claimed in claim 1 wherein Shilajit is a purified powder. 6) A herbal composition as claimed in claim 1 wherein the ratio of Withania somnifera and Shilajit is preferably 1:1. 7) A herbal composition as claimed in claim 1 wherein the composition is a capsule, tablet or syrup. 8) An herbal composition substantially as herein described with reference to the foregoing examples and accompanying drawings.

Documents:

3-del-2002-abstract.pdf

3-del-2002-claims.pdf

3-del-2002-correspondence-others.pdf

3-del-2002-correspondence-po.pdf

3-del-2002-description (complete).pdf

3-del-2002-drawings.pdf

3-del-2002-form-1.pdf

3-del-2002-form-19.pdf

3-del-2002-form-2.pdf

3-del-2002-form-26.pdf

3-del-2002-form-3.pdf

3-del-2002-form-5.pdf

3-del-2002-petition-138.pdf