Title of Invention	A PROCESS FOR ISOLATION OF (-)_OLIVIL FROM A NEW SOURCE STEREOSPERMUM PERSONATUM
Abstract	The process of present invention provides for isolation of (-)-Olivil from a new natural source Stereospermum personatum wherein the yield of said (-)-Olivil is higher than already known source.

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A process for isolation of (-)- Olivil from a new source Stereospermum personatum useful as antioxidant. This invention relates to the isolation of compound namely (-)-0/w7 from a new plant source, Stereospermum Personatum in good yield. It is proved to be useful and better antioxidant molecule than the presently used medicinally important lipophilic antioxidants Prubucol and a-Tocopherol. It may have better therapeutic potential in inflammatory disease conditions, atherosclerosis, diabetic complications, cancer, hepatotoxicity and variety of disease conditions mediated through or fostered by oxidative stress and / or overt oxidative burden due to increased generation or under scavenging of free radicals. There is a considerable amount of epidemiological evidence indicating association between diet rich in fruits and vegetables and a decreased risk of cardiovascular disease and certain forms of cancer. It is generally assumed that the active principles contributing to these protective effects are nothing but primarily, the antioxidant phytochemicals. Research in the past decades have accumulated enough evidence to show the beneficial effect of free-radical scanvengers/antioxidants as antimutagenic, antinflammatory, antiamerosclerotic, antidiabetic, antihepatotoxic, antiageing and hi a variety of neurological disorders. The search for new antioxidant principles is becoming therefore, essential to improve the pharmacological treatment of pathalogical conditions related either due to free radical / oxidative damage or due to imbalance between antioxidant/oxidant homeostasis such as cataract, rheumatic diseases, atherosclerosis, Alzheimer's disease and other neurodegenarative conditions. The pharmacological approaches therefore have focused on the search of potential resources rich in antioxidant principles. The medicinal importance of plants bearing rich proportion of antioxidant principles is therefore becoming hot item. Stereospermum personatum is a medicinal plant used in traditional Indian System of Medicine and is widely advocated in the preparations for diuretic, lithontriptic, expectorant, cardiotonic, aphrodisiac, appetizer, anti-inflammatory, antibacterial, dyspepsia, diarrhea, renal and vesical calculi, cough, asthma, hyperdipsia, haemorrhoids and hyperacidity disease conditions( Indian Medicinal Plants, Vol 5 p 192). It is further reported to possess antibacterial, antifungal, hypoglycaemic activity, and against p388 lymphocytic leukemic cells (Ind. Jour, of Exp. Biol., 1971,9, 100). Hence, it becomes pertinent to look for the molecules possessing such important biological properties. In this connection, the phytochemical investigation of Stereospermum personatum has been taken up. The applicants made efforts to the isolation of (-)-OIMI in good yield. (-)-Olivil is known to possess several activities, which are shown in Table. 1 Table. 1 (Table Removed) The main object of the invention is to provide a process for preparation of (-)- 0//V/7 from a new natural source namely Stereospermum personatum useful as antioxidant. Another object is to provide a new source for obtaining (-)-Olivil in good yield. Yet another object of the invention is to provide a process for producing (-)- 0//V/7 from a plant source namely Stereospermum personatum. Yet another object of the invention is relates to use of (-)-Olivil as an antioxidant. Still another object of the present invention is to use (-)-Olivil for the preparation of a novel composition containing (-)-OIMI and an additive, which is used as an antioxidant. Accordingly, the present invention provides a process for isolation of (-)-Olivil from a new source Stereospermum personatum characterized in that the yield of said (-)-OIMI is higher than already known source , said process comprising the steps of: a) extracting the dried wood powder of Stereospermum prsonatum with hexane; b) further extracting the residue from step (a) with chloroform; c) concentrating the chloroform solution from step (b) under vacuum; d) absorbing the dark brown extract on a silicagel (60-120 mesh) and loaded on silicagel (60-120mesh) column (4 cm dia to height of 100 cm); e) eluting the column with chloroform methanol gradient, and f) collecting the eluted fraction with 4% methanol in chloroform and concentrating the fraction to obtain pure (-)-Olivil. In an embodiment of the present invention, the present invention, the solvents used are selected from hexane, chloroform and methanol. In another embodiment of the present invention, the yield of (-)-Olivil obtained is about 0.073% of the plant dried material. In still another embodiment of the present invention, the (-)-Olivil may be used as an antioxidant or free radical scavenger for animals and human beings. In yet another embodiment relates to use of (-)-Olivil for the manufacturing of a composition useful as an antioxidant. In yet another embodiment of the present invention, (-)-Olivil is administered orally. Accordingly, the invention provides a novel composition containing (-)-Olivil and useful as antioxidant. The invention further provides a method for the isolation of (-)-Olivil froma new plant source namely Stereospermum Personatum. In accordance with this invention, it has been found that (-)-Olivil is isolated from a new source, Stereospermum Personatum in significant yield and it has been found that (-)-Olivil is isolated from a new source, Stereospermum Personatum in significant yield and it has been found that (-)-Olivil show antioxidant property. Antioxidant compounds recently have attracted the attention due to their broad spectrum of activities in disorders of multiple origin viz., coronary heart disease, cancer, diabetes, reheumatic disorders and inflammatory conditions where free radicals play important role. Much attention is being directed now to harness and harvest the antioxidant compounds from natural resources. The compound (-)-Olivil is used in pure form. Hence, the usage may be more advantageous than a mixture of compounds having similar properties, which are in current use. It is also important to note that the process of isolation of (- )-Olivil is highly economical. The present invention provides a useful source, Stereospermum personatum, comprising antioxidant principle of which (-)-Olivil is isolated as pure and potent antioxidant molecule. (-)-Olivil has been compared with existing pharmacologically / therapeutically accepted antioxidant Probucol and alpha-tocopherol. It is found that (-)-Olivil is better than the above mentioned reference drugs and hence may be used with pharmaceutically / cherapeutically acceptable additives. (-)-Olivil may be effective in much less amount than the reference drugs mentioned above. One embodiment of the present invention relates to a composition useful as antioxidant, said composition comprising an effective amount of (-)-OliviI in combination with or associated with a pharmaceutically acceptable additives. In another embodiment of the invention, the pharmaceutically acceptable additive is selected in such a manner that it does not affect or interfere with the efficacy of (-)-Olivil. In still another embodiment of the invention, the additive is selected from nutrients such as carbohydrates, sugar, proteins, fats and pharmaceutically acceptable carrier. In an embodiment of the present invention, the ratio of (-)-Olivil to the additive is in the range between 0.1:10 to 3:10, preferably 0.4:10 to 2:10. In yet another embodiment of the invention, the amount of (-)-Olivil administered is in the range between 300 mg to 400mg per dose at least twice a day In yet another embodiment of the invention, the composition is administered through oral route in the form of tablets, capsules, syrup or powder. Yet another embodiment of the invention relates to a method for the treatment of free radical scavenging, comprising the steps of administration of an effective amount of (-)-Olivil to a subject in need thereof preferably, (-)-Olivil is administered in the range of 300 mg- 400mg/dose, twice a day. Stereospermum personatum hence is a new source for (-)-Olivil and its presence in this plant in good yields makes this invention more important. A comparison of yield (-)-Olivil from different plants is given in Table.2 Table.2 (Table Removed) The present invention embodies isolation of (-)-Olivil, as antioxidant principle from a new plant source and identify its free radical scavenging property compared with medicinally important antioxidant drug molecules. Brief description of the accompanying drawing: Figl shows antioxidant (Anti-free radical, DPPH) activity of (-)-Olivil from Stereospermum personatum and reference compounds Fig 2 represents the structure of (-)-Olivil. In our copending patent application no. NF251/2001 we have claimed a pharmaceutical composition useful as antioxidant using (-)-Olivil. The invention is further described in the following examples that are given by the way of illustration and therefore should not be construed to limit the invention in any manner. Example 1 Experimental protocol: A process for the isolation of above lignans. The dried wood powder of Stereospermum personatum (3 Kg) was loaded on a Soxhlet apparatus. The powder was first extracted with hexane. The residue from the extraction of hexane was further extracted with chloroform. The chloroform solution was concentrated under vacuum. The dark brown extract (22 g) was adsorbed on silica gel (60-120 mesh) and loaded on silica gel (60-120 mesh) column. (4 cms diameter to a height of 100 cms). The column is subjected to elution with chloroform methanol gradient. The chloroform- methanol gradient is so selected to obtain specific fraction and thereby the desired compound. In the present case, the fractions eluted at 4% methanol in chloroform are collected separately and concentrated. The above fractions are subjected to further purification using silica gel column (>200mesh, 2.5 cm dia and 50 cm length) using chloroform methanol gradient. The eluent at 4% methanol in chloroform gave pure (-)-Olivil (2.2 g). The spectrochemical data of (-)-Olivil are given below: 1. Molecular formula: 2. 'HNMR: 5 2.45(1H, m), 2.9(1 H, d), 3.05(1H, d), 3.65(1H, d), 3.95-3.80(3H, m), 3.85(6H, s), 4.7(1H, d ), 5.6 (Ar-OH, s), 5.65(Ar-OH, s), 6.75-6.90(5H, m), 7.1(1H, s). 3. 13CNMR: 5 39.19(C-7'), 55.80(2xOCH3), 59.42(C-8'), 59.91(C-9), 77.79(C-9'), 81.18(C-8'), 83.64(C-7), 109.60(C-2'), 113.50(C-2), 1 14.46(C-5,),lI4.60(C-5), 119(C- 6,),I22.83(C-6), 128.56(G-l),133.64(C-r), 144.56(C-4,), 1 45.47 (C-4), 146.82(c-3'), 147.18(C-3). 4. MS: 376(M+) 5. IR: 3401 crn1 Example 2 In vitro evaluation of free radical scavenging antioxidant potency: Antioxidant activity of the compounds was tested for its capacity/potency to scavenge most widely used free radical, l,l-diphenyl-2-picryl hyrazyl radical (DPPH). The well-accepted and tested antioxidants namely probucol and a-tocopherol were taken as reference compounds. 1mg/ml DMSO concentration of the compounds were prepared and subsequently, diluted to lower concentrations with DMSO. 200 µl of test compounds were reconstituted to 1 ml in tris-HC1 buffer (pH 7.4). Equal volume of 500 µM of DPPH radical dissolved in ethanol was reacted with this. After incubation for 45 minutes in dark, the absorbency at 517 nm was recorded. Percent radical scavenging activity was calculated accordingly. All the readings were taken in triplicate. Results (Fig.l and Table 3) shows that compound under consideration possess potent antioxidant/free- radical scavenging property. Table 3 50 % Radical scavenging concentration of compounds (Table Removed) We Claim 1. A process for isolation of (-)-Olivil from a new source Stereospermum personatum characterized in that the yield of said (-)-Olivil is higher than already known source , said process comprising the steps of: a) extracting the dried wood powder of Stereospermum personatum with hexane; b) further extracting the residue from step (a) with chloroform; c) concentrating the chloroform solution from step (b) under vacuum; d) absorbing the dark brown extract on a silicagel (60-120 mesh) and loaded on silicagel (60-120mesh) column (4 cm dia to height of 100 cm); e) eluting the column with chloroform methanol gradient, and f) collecting the eluted fraction with 4% methanol in chloroform and concentrating the fraction to obtain pure (-)-Olivil. 2. A process as claimed in claims 1 wherein, the yield of (-)-Olivil is 0.073 percent of the dried plant material. 3. A process as claimed in claims 1 - 2, wherein (-)-Olivil is used as an antioxidant or free-radical scavenger. 4. A process for isolation of (-)_OIMI from a new source Stereospermum personatum substantially as herein described with reference to the examples and drawing accompanying this specification.

Documents:

857-del-2001-abstract.pdf

857-del-2001-claims.pdf

857-del-2001-correspondence-others.pdf

857-del-2001-correspondence-po.pdf

857-del-2001-description (complete).pdf

857-del-2001-drawings.pdf

857-del-2001-form-1.pdf

857-del-2001-form-18.pdf

857-del-2001-form-2.pdf

857-del-2001-form-3.pdf

857-del-2001-petition-137.pdf