Title of Invention

A NEUROPROTECTIVE PHARMACEUTICAL COMPOSITION

Abstract

The invention relates to a neuroprotective pharmaceutical composition comprising Curcuma Longa rhizomes and/or, Azaradicta Indica leaves and/or, atleast one plant material selected from Emblica officinalis fruits and Tinospora Cordifolia bark in combination with magnesium oxide and/or Taurine, Tyrosine and lauric acid. The composition shows significantly high stimulatory and inhibitory effect on Na<sup>+</sup>, K<sup>+</sup>, ATPase and HMG CoA reductase respectively than when each of the components is used individually.

Full Text

This invention relates to a neuroprotective pharmaceutical composition. This composition is particularly but not exclusively useful in the treatment of the following diseased conditions. 1. Primary generalised epilepsy 2. Schizophrenia 3. Parkinson"s disease 4. Multiple sclerosis 5. Refractory CNS glioblastoma 6. Syndrome X with vascular disease 7. Neuronal aging and Dementia of the Alzheimer"s type 8. Down"s syndrome 9. Acquired immunodeficiency syndrome. Modem medicines for treatment of these disorders are not satisfactory and are often found to produce undesirable side effects. Object of this invention is to develop a safe and effective medicine which can be effectively used to ameliorate these disorders with minimum or negligible side effects. Research work carried by us over a period of years has shown that patients of the disorders/conditions mentioned above show:- 1. Decrease in the activity of a cell membrane bound enzyme known as Na+-K+ ATPase which plays an important role in the transport of cations in and out of cells. 2. There is increase in the concentration of digoxin in the serum. Digoxin is a sterodial glycoiside, now known to be synthesised by the hypothalamus whose main physiological action is inhibition of Na+-K+ ATPase activity. 3. The increase in the concentration of digoxin in the serum is associated with decrease in the activity of Na+-K+ ATPase and therefore increased digoxin in these disorders is the cause of the inhibition of this enzyme. 4. Digoxin is synthesised in the hypothalamus by what is known as the mevalonate or isoprenoid pathway which starts from acetyl CoA and this pathways synthesises a number of substances which include cholesterol, coenzyme Q10 and dolichol in addition to digoxin. A major controlling factor in the operation of this pathway is the activity of an enzyme known as HMG CoA reductase. 5. An inhibition of cell membrance Na+-K+ ATPase results in an increase in intracellular calcium levels and a decrease in intracellular magneisum concentration. 6. A decrease in serum magnesium has been observed by us in all these disorders. 7. Various reports suggest that increase in intracellular calcium and decrease in intracellular magnesium are involved in the pathogenesis of the disorders/conditions mentioned above. We therefore felt that if the digoxin level can be kept low so that the enzyme Na -K ATPase exerts its normal action, it can go a long way in the treatment of these disorders. Attention was therefore focussed to find agents which can act in this direction. We screened a large number of medicinal plant extracts and various others substances for their effect on- 1. activity ofmembrane Na+-K+ ATPase. 2. Activity of HMG CoA reductase, an inhibition of which would decrease digoxin level. These studies were carried out in experimental animals like rats and those which were found to be effective in stimulating the activity of membrane Na+-K+ ATPase and decreasing the activity of HMG CoA reductase were tested in detail for their systemic toxicity. The following were found to be effective in stimulating Na+-K+ ATPase and inhibiting HMG CoA reductase and which were without any toxic effects even at high doses. 1. Extract/whole powder of curcuma longa rhizomes (Haldi or turmeric). 2. Extracy/whole power of Azaradicta Indica leaves (neem). 3. Extract/whole powder of Emblica Officinalis (Amla fruit). 4. Extract/whole powder of the stem of Tinospora Cordifolia baric (Amruta). 5. Magnesium oxide. 6. Taurine. 7. Tyrosine. 8. Laurie acid. Each of these substances has a stimulatory effect on Na+-K+ ATpase and a inhibitory effect on HMG CoA reductase. However these effects vere significantly more when all of these were used in combination. Method of preparation of dried solvent extract for formulation: Fresh or dry material in each case was extracted with 90% methanol and the extract was concentrated in vacuum to remove mose of the methanol. The concentrate was then extracted with n-hexane and the aqueous layer was concentrated and dried at low temperature in a lyopbiliser to obtain a powder. Preparation of the powder of the whole material: The material was sun dried and powdered to get powder of 100-200 micron size. Preparation of the fonnulation: The following formulation in the concentration mentioned was made. Cone. 1. dry extract of Curcuma Longa - A 250 mg 2. dry extract of Azardicta Indica - B 250 mg 3. dry extract of Emblica Officinalis - C 250 mg 4. dry extract of Tinospora Cordifoiia - D 250 mg 5. Magnesium oxide (IP grade )-E 1000 mg 6. Tyrosine (LP. grade )- F 500 mg 7. Taurine (IP - grade) - G 100 gm 8. Laurie acid (IP grade)-H. 250 mg Components A, B,C & D (1 to 4) were mixed to form part I and components E,F,G & H ( 5 to 8) were mixed separately to form Part II. The clinical trials, details below in example I were carried out with this fonnulation. However the range of concentration of the various components which showed effect is given below. 50 mg to lOOOmg in the case of dry extracts of A, B, C and D. 250 mg to 2gm of Magnesium oxide. 100 mg to 10(X)mg of tyrosine 25 mg to 500 mg of taurine. 25 mg to 1 gm of lauric acid. It is to be noted that at least one of the extracts selected from Curcuma Longa and Azaridicta Indica must be present in combination with the substances of non-plant origin. A combination of components represented under 1 to 8 or A to H is found to exhibit maximum potency whereas a combination of 1,3 and 4 or 2,3 and 4 with components of 5-8 also are found efifective. A combination of either 1(A) alone with 5-8 or 2 (B) alone with components 5-8 is also effective. Significant effect is also observed for a combination of 1 to 4 with 5 alone. A combination 1 and 5 or 2 and 5 is also effective. This invention relates to a neuroprotective pharmaceutical composition comprising Curcuma Longa rhizomes, and/or Azardicta Indica leaves and/or at least one plant material selected from Emblica Officinalis fruits and Tinospora cordifolia bark in combination with Magnesium oxide, and/or Taurine, Tyrosine and lauric acid. The extracts of the plant material may be prepared with methanol or any other polar solvent and apart from hexane, any other non-polar solvent can also be used for the extraction of the concentrate. The extracts can also be aqueous extracts of the plant material, made by boiling with water and concentration of the extract to dryness. EXAMPLE 1 Clinical Trials: We carried out clinical trials with this formulation in patients with 1. Primary generalised epilelsy 2. Schizophrenia 3. Parkinson"s disease 4. Multiple sclerosis 5. Refractory CNS glioblastoma 6. Syndrome X with vascular disease 7. Neuronal aging and Dementia of the Alzheimer"s type 8. Down"s syndrome 9. Acquired immunodeficiency syndrome. Each patient was administered Part I and II of formulation daily. The patients were assessed before treatment was started clinically and by all required laboratory investigations. The duration of treatment ranged from 6 months to 2 years. Their condition was assessed during treatment and after treatment clinically and using all necessary laboratory investigations. We found that in the cases tried, the formulation showed significant curative effect. None of the substances mentioned in the formulation has been used before either singly or in combination for the purpose for which they are described to be used. The invention will now be illustrated with reference to the following typical examples. 1. Refractory Epilepsy; Male aged 15 years with primary generalised epilepsy. This patient was refractory to treatment and was on a combination of carbamazepine - 1200 mg/day, sodium valproate ~ 1200 mg/day and dilantin sodium 800 mg/day. The seizure frequency at the start of therapy was 12 episodes/day. The formulation was started with the dose of carbamazepine, sodium valproate and dilantin sodium reduced to half the respective dose in the 1 month and 1/4* respective dose in the 2" month and withdrawn from the 3"* month onwards. The treatment duration was I year. At the end of 1 year the seizure frequency reduced to 2/month. There were no side effects noticed. 2. Refractory schizophrenia Female aged 25 years with refractory schizophrenia of 3 years duration. The patient was on risperidone - 9mg/day and clozapine - 75 mg/day. The formulation was started and the doses of risperidone and clozapine reduced to half the respective dose for 1 month, YA the respective dose for 2 month and completely withdrawn from 3"* month onwards. The duration of the treatment was 1 year. The scoring values at the start and end of therapy was as follows. Score 3. Refractory Parkinson"s Disease Male aged 60 years with idiopathic Parkinson"s disease. He was on syndopa (L-dopa + Carbidopa) - 1.5 mg/day, Bromocryptine - 7.5 mg/day and pacitane - 12 mg/day. The formulation was started with syndopa,, bromocryptine and pacitine reduced to half the respective dose in the first month, and VA of the respective dose in the 2" month. These drugs were withdrawn from 3"** month onwards. Duration of the treatment was 1 year. The UPDRS ratings scales were used which have the following parameters. mentation/behaviour/mood i. activities in daily living ii. motor examination V. complication of therapy Based on UPDRS scales the patient showed significant improvement. No side effects were noticed during treatment. 4. Refrm;tory Multiple sclerosis Female aged 28 years diagnosed as having multiple sclerosis based on Poser"s criteria. The patient was on routine immunosuppressive therapy with prednisolone - 60 mg/day and azathioprine - 100 mg/day. She was put on the formulation with the respective doses of prednisolone and azathioprine reduced to half the respective dose in the 2"* month and totally withdrawn from the 4* month onwards. The duration of the treatment was 2 years. The parameters before starting therapy was 1. Relapse rate - 6 relapses/year 2. Activity of daily living scale - cannot carry out the activities of daily living and was bed ridden - grade IV 3. MRI scan with gadolinium contrast showed active lesions. The parameters after therapy for 3 years were !. Relapse rate - 0 per year. No relapses were noticed. 2. Activity of daily living scale - could perform activities of daily living without help. 3. MRI scan with gadolinium contrast repeated at 6** month, 1 year, V/i years, 2 years, 2 Vz years and 3 years showed no active lesion. 5.Refractory CNS glioblastoma Male aged 34 years old with massive left frontoparietal glioblastoma with midline shift. The patient had already undergone the routine radiotherapy and taken the chemotherapy course. He was put on the formulation I. The duration of the treatment was 5 years. After 2 years treatment repeat MRI scans showed a 60% quantitative reduction in tumour size. After 4 years of treatment repeat MRI scans showed a further 25% (total of 85% from initial size) reduction in tumours size. Before starting treatment based on his clinical and MRI findings he was prognosticated to have a 3 months survival. After treatment with the formulation he had a 4 year survival. 6. Acquired immunodeficiency syndrome Male aged 35 years diagnosed as having acquired immunodeficiency syndrome. He was positive for HIV by both elisa and western blot. He had generalised lymphadenopathy and hepatosplenomegaly had regressed. His weight w 52 Kg. The initial CD4 count was 110 cells/cum m. He was put on formulation. The treatment duration was 1 year. After 6 months of therapy the CD4 count increased to 400 cells/cum m and after 1 year of therapy to 500 cells/ cum m. The weight increased to 65 Kg. His lymphadenopathy and hepatosplenomegaly had regressed. The formulation was effective in his case. 7. Syndrome X Female aged 52 years with freshly diagnosed non-insulin dependent diabetes mellitus, obesity, hypertension, hypertriglyceridemia, unstable angina and recurrent episodes of TIA. She was put on formulation . The duration of treatment was 1 year. Parameters before starting treatment were as follows: 1. fasting blood sugar - 186 mg% post prandial blood sugar - 420 mg% 2. Serum triglycerides - 600 mg% 3. Episodes of unstable angina - 6/month ECG showed inferolateral ischaemia 4. Episodes of transient ischaemic attack of the MCA territory - 3/year 5. Weight of the patient - 85 kg. 6. Insuhn requirement - was on 45 units of lente insulin daily. The treatment duration was 2 years: Parameters after treatment were as follows. 1. Fasting blood sugar - 96 mg% post prandial blood sugar - 142 mg% 2. Serum triglycerides - 120 mg% 3. Episodes of unstable angina- nil/month ECG showed no changes 4. Episodes oftransient ischaemic attack ofthe MCA territory- nil/year 5. Weight of the patient - 60 kg. 6. Insulin requirement - was halved to 25 units of lente insulin daily in the first month, 15 units of lente insulin daily in the second month and withdrawn totally by the third month. There are no side effects for treatment. 8. Neuronal aging and Dementia ofthe Alzheimer"s type: Male aged 72 years diagnosed as having Alzheimer"s disease by NINDS criteria. The mini-mental status examination before therapy gave a score of 6. He was dependent on others for his activities of daily hving. He was put on the formulation . The duration of treatment was 2 years. The mini-mental status examination score at the end of 2 years of treatment was 26. He was independent with regard to the activities of daily living. The treatment was without any side effects. 9. Down"s syndrome - Trisomy 21: Male aged 9 years had severe mental retardation with a diagnosis of trisomy 21. His IQ assessment gave a value of 20 before therapy. The patient was put on the formulation for 2 years. The IQ assessment at the end of the therapy gave a value of 55. There were no side effects for the therapy. Patient population included in the large scale trial: These are typical examples of a large number of patients tried in each case. The number of patients included in the trial are as follows: 1. Primary generalised epilepsy - 50 patients. 2. Schizophrenia - 50 patients 3. Parkinson"s disease-50 patients 4. Multiple sclerosis - 25 patients 5. Refractory CNS glioblastoma - 10 patients 6. Syndrome X with vascular disease - 50 patients 7. Neuronal aging and Dementia of the Alzheimer"s type - 25 patients 8. Down"s syndrome-10 patients 9. Acquired immunodeficiency syndrome - 15 patients EXAMPLE 2 Details of the trial in the disorders discussed above were the same except that the formulation containing the whole powder of medicinal plants A, B, C and D was used with E, F, G and H. The formulation had the following composition. The herbal components were mixed to form one part (I) and the other components (magnesium oxide, tyrosine, taurine and lauric acid) were mixed to form the second part (II). This composition was used for the clinical trial. Similar results were obtained in the clinical trial. The formulation was 25% less effective when compared to the formulation containing dry extracts. The range of concentration of whole powder of the plant material is 500 mg to 5.0 gm. EXAMPLE 3 Details of the trial were the same except that D was omitted from the formulation. Similar results were obtained. The formulation was 25-30% less effective. EXAMPLE 4 Details of the trial were same except that C was omitted from the formulation used in example 1. Similar results were obtained. The formulation was 10-15% less effective. EXAMPLE 5 Details of the trial were same except that B was omitted from the formulation used in example 1. Similar results were obtained. The formulation was 15-20% less effective. EXAMPLE 6 Details of the trial were same except that B, C & D were omitted from the formulation used in example 1. Similar results were obtained. The formulation was 30-40% less effective. EXAMPLE 7 Details of the trial were same except that A, C and D were omitted from the formulation used in example L Similar results were obtained. The formulation was 30-40% less effective. EXAMPLE 8 Details of the trial were same except that tyrosine was omitted from the formulation used in example 1. Similar results were obtained. The formulation was 20-25% less effective. EXAMPLE 9 Details of the trial were same except that magnesium oxide was omitted from the formulation used in example 1. Similar results were obtained. The formulation was 40-50% less effective. EXAMPLE 10 Details of the trial were same except that taurine was omitted from the formulation used in example 1. Similar results were obtained. The formulation was 20-25% less effective. EXAMPLE 11 Details of the trial were same except that lauric acid was omitted from the formulation used in example 1. Similar results were obtained. The formulation was 10-15% less effective. EXAMPLE 12 Details of the trial were the same except that D was omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 25-30% less effective when compared to example 2. EXAMPLE 13 Details of the trial were the same except that C was omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 10-15% less effective when compared to example 2. EXAMPLE 14 Details of the trial were the same except that B was omitted fixjm the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 15-20% less effective when compared to example 2. EXAMPLE 15 Details of the trial were the same except that B, C & D were omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 30- 40% less effective when compared to example 2. EXAMPLE 16 Details of the trial were the same except that A, C & D were omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 30-40% less effective when compared to example 2. EXAMPLE 17 Details of the trial were same except that tyrosine was omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The tbrmulation was 20-25% less efiective when compared to example 2. EXAMPLE 18 Details of the trial were same except that magnesium oxide was omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 40-45% less effective when compared to example 2. EXAMPLE 19 Details of the trial were same except that taurine was omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 20-25% less effective when compared to example 2. EXAMPLE 20 Details of the trial were same except that lauric acid was omitted from the formulation of whole powder mentioned in example 2. Similar results were obtained. The formulation was 10-15% less effective when compared to example 2. WE CLAIM: 1. A neuroprotective pharmaceutical composition comprising Curcuma Longa rhizomes and/or Azaradicta Indica leaves and/or atleast one plant material selected from Emblica officinalis fruits and Tinospora Cordifolia bark in combination with magnesium oxide and/or Taurine, Tyrosine and lauric acid. 2. The composition as claimed in claim 1, wherein whole fine powder of the plant material or extracts thereof are used. 3. The composition as claimed in claim 2, wherein said extracts are dried solvent extracts obtained by solvent extraction of said plant material. 4. The composition as claimed in claim 1 or 2 wherein the said extracts are dried water extracts obtained by boiling the material with water and drying the water extract. 5. The composition as claimed in claim 3, containing 50 mg to 1000 mg of the dried solvent extracts of each plant material. 6. The composition as claimed in claim 2, containing 500 mg to 5 gm of dry plant material. 7. The composition as claimed in claim 4 containing 50 - 1000gm of the dry water extract. 8. The composition as claimed in claims 1 to 5, containing 250 mg to 2 gm of Magnesium oxide. 9. The composition as claimed in claims 1 to 6, containing 100 mg to 1 gm of tyrosine. IC The composition as claimed in claims 1 to 7 containing 25 mg to 500 mg of taurine. 11 The composition as claimed in claims 1 to 8 containing 25 mg to 1 gm of lauric acid. 12. The composition as claimed in claim 3, wherein said solvent extract is obtained by extraction of the plant material with a polar solvent such as methanol, ethanol or acetone, concentration of said extract and subsequent extraction of said concentrate with a non- polar solvent such as hexane, chloroform and petroleum ether. 13. A neuroprotective pharmaceutical composition substantially as herein described.

Documents:

0720-mas-2000 abstract duplicate.pdf

0720-mas-2000 abstract.pdf

0720-mas-2000 claims duplicate.pdf

0720-mas-2000 claims.pdf

0720-mas-2000 correspondance others.pdf

0720-mas-2000 correspondance po.pdf

0720-mas-2000 description(complete) duplicate.pdf

0720-mas-2000 description(complete).pdf

0720-mas-2000 form-18.pdf

0720-mas-2000 form-26.pdf

0720-mas-2000 form-3.pdf

0720-mas-2000 form-4.pdf