Title of Invention

"A CONVENIENT METHOD FOR THE LARGE-SCALE ISOLATION OF GARCINIA ACID"

Abstract

A process for the isolation of Garcinia acid from the fresh or dried rinds of the fruits of Garcinia indtca, Garcinia cambogia, Garcinia atroviridis, comprising the steps of, in sequence : a) subjecting the rinds to extraction to form an extract. b) Adding a solvent to the extract to remove pectin and form a filtrate c) Converting the filtrate to form an alkali salt d) Neutralizing the alkali salt with an acid, followed by evaporation, to form a concentrate e) Purifying said concentrate using a solvent to remove inorganic matter as impurities, to form a second filtrate f) Concentrating the second filtrate to ytekl a crude Gardnia acid g)Recrystallizing the crude to form pure crystals of Gradnia acid.

Full Text

A CONVENIENT METHOD FOR THE LARGE-SCALE ISOLATION OF GARCINIA ACID. Background of the invention: This invention is related to an improved and efficient process for the large-scale isolation of (-)-hydroxycitric acid or Garcinia acid (2S,3S-dihydroxy 1,2,3-propanetricarboxylic acid) from fresh or dried rinds of the fruits of Garcinia cambogia, Garcinia indica and Garcinia atroviridis. Garcinia acid is widely used as an important ingredient in many pharmaceutical formulations REFERENCES: 1. US patent No. 4005086 dated 25/1/77; 2. US patent No. 4006166 dated 1/2/77; 3. US patent No. 4007208 dated 8/2/77; 1. US patent No. 5536516 dated 1.6/7/96; 5. US patent No. WO 9605741 Al 960229; 6. US patent No. WO 9636585 Al 961121; 7. CA 86, 1977, 186629r; 8. CA85, 1976, 4153 Ix; 9. CA 87, 1977, 195626k; 10. CA 96, 1982, 30421n). The compound can be used as a potential precurser for the syntheses of many optically active natural products References . a. Tetrahedron Letters Vol. 25 pp. 4491-4494, 19X4; b. Tetrahedron Vol. 43, No. 19, pp 4497-4506, 1987; c. Tetrahedron Vol. 38, No. 15, pp 2377-2394, 1982; d. Tetrahedron Vol. 34, pp 1449-1452, 1978; c. J. Org. Chem. 63, 2385-2388, 1998; f. JOS Chem. Comm. pp 711, 1973; 7. g. Org. Chem. 58, 2725-2737, 1993; h. Tetrahedron Vol. 31, pp 3011-3012, 1975; i. Tetrahedron Letters Vol. 22, No. 52, pp 5271-5274, 1981; j. Tetrahedron Letters Vol. 23 No. 48 pp 5051-5054, 1982). However Garcinia acid, in the optically pure form, is not available in the market. This has resulted in the limited use of this compound in the synthetic front. The nonavailability of this compound in the open market is due to the absence of any commercially viable large-scale manufacturing process. The present method describes an economic, commercially viable, cost effective process for the large-scale isolation of Garcinia acid (la). Existing methods : a. The method described by Y. S. Lewis and S. Neelakanfan ( Phytochemistry Vol. 4, 1965, pages 619-625) involves autoclaving the dried rinds of the fruits of Garcima cambogia with water followed b)' the concentration of the filtered extract. Two volumes of alcohol is added to the concentrate to remove pectin and the filtrate is neutralised with alkali. The heavy liquid separated is washed several times with 60% aqueous alcohol to gel alkali salt of the acid. The aqueous solution of the alkali salt is then passed through cation exchange resin to get(-)-hydroxy citric acid, iiluent on concentration yields crude lactone of the acid which is dried on water bath for several hours before drying in a vacuum oven for (en hours at 100° C. Finally the product is kept in a desiccater for several days to get a crude crystalline mass which is recrystallised from ether. b. US patent No. 5536516 dated 16/7/96 provides a method for enriching the content of (-)-hydroxycitric acid by subjecting the crude Garcinia rind extract to ion-exchange chromatography. c. US patent No. 5656314 and Patent No. WO 9605741 A1 960229 describes the preparation of a mixture of (-)-hydroxycitric acid , its lactone and citric acid. d. The method reported by Per. M. BolL, Else Sorenseii and Erik BaHeu (Acta Chem. Scand 23 pp. 286-293, 1969) for the synthesis of Garcinia acid is a synthetic procedure. This involves oxidation of tams-aconitic acid using silver chlorate and osmium tetroxide followed by purification of the reaction mixture by preparatory layer chromatography in the milligram-scale. The oily material obtained is dried over P2O5 aad recrystallised. Drawbacks of prior arts : The main drawbacks of the existing method "a" are as follows. 1. It employs expensive tedious ion-exchange chromatography. 2. It is useful only for preparations in the milligram or gram scale. 3. Crystallisation of the product takes several days. 4. It is not at all suitable for large-scale isolation of the compound. The drawbacks of the existing method "b" are listed below. 1. This method only enriches the content of (-)-hydroxycitric acid in Garcinia rind extract. It does not provide any means for the isolation of (-)-hydroxycitric acid lactone crystals. 2. It is suitable only for milligram or gram scale preparations. The drawbacks of the existing method "c" are listed below. Existing method "c" describes only a method for preparing a concentrate containing (-)-hydroxycitric acid , its lactone and citric acid. This does not provide a process for the isolation of pure Garcinia acid. The drawbacks of existing method "d" are given below. 1. This method cannot be scaled up. 2. It involves the use of very expensive reagents. Advantages of the new process : Principally the present invention is totally an alternative method for the Isolation of Garcinia acid avoiding ion-exchange chromatography. This process provides a simple and inexpensive method of obtaining crystals of Garcinia acid in the optically pure form, from natural sources. The process is more efficient than all listed previous methods and it involves the use of cheap and reusable solvents, simple extraction methods and is less time consuming. This method can be used for the production of Garcinia acid in the large scale. Crystallisation procedure described here is very fast and simple. ' This assumes importance considering the potential of mis compound in the pharmaceutical as well as synthetic front. Detailed description of the new invention : The present invention involves the following steps. 1. The dried or fresh rinds of the fruits of Garcinia cambogia, Garcinia indica or Garcinia atroviridis are cut into small pieces and soaked in boiling water for 1.0-20 hours. The water extract is collected. The extraction Ls repeated a few times. The combined extracts are evaporated to a syrup(A).The soaking for longer time is avoided when fresh fruits are used. The water extract is also prepared by autoclaving the fresh or dried fruits. 2. To the syrup (A), sufficient quantity of alcohols like methanoL ethanol or butanol Ls added to remove pectin completely. The filtrate Ls concentrated to a syrup (13). Alternatively' the fruit rinds are subjected to exhaustive soxhlet extraction using alcohols like methanol, ethanol or butanol to get pectin free extract. Recovered solvent is preserved for use at. a later stage of the process. 3. After making syrup (B) alkaline by adding sufficient quantity of alkali at elevated temperature, alcohols like methanoL ethanol or butanol is added to the solution. Separated thick syrup (lower layer) is washed several times with aqueous alcohols like rnethanoi, ethanol or butanol of varying proportions to get a paste of alkali salt (C). Recovered alcohol from step 2 is used for preparing aqueous alcohol. 4. The alkali .salt (C) is neutralised with a mineral acid like hydrochloric acid arid Ls evaporated to get a concentrate (D). 5. The concentrate (D) is triturated with sufficient quantity of an organic solvent like acetone, ether or methanol to precipitate inorganic matter and the filtrate upon concentration yielded crude Garcinia acid (E) 6. The crude acid (E) up on recrystallisation from solvents like acetone, methanol, ethanoi, chloroform dichloroethane or their appropriate mixtures gives pure crystals of Garcinia acid (la). Yield is 5-10% of the weight of the dried fruit rinds. The purity of the product is confirmed by spectroscopic data and other physical data. The values of IR spectrum, melting point and optical rotation are comparable with the reported data. The reported proton nmr values of Garcinia acid dates back to 1969. We have observed some anomalies in the proton nmr spectrum reported for the compound. The observed and reported values are given below. (Table Removed) Literature survey reveals that there are no data available on 13C nmr spectrum and mass spectrum of Garcinia acid. We have recorded '"'(.' nmr spectrum and mass spectrum of the compound and the values are given below. instrument used for 13C nmr spec (rum was a Brucker AMX 400 (400 MHz) spectrometer. 13C nmr spectrum : 5 174.03, 171.06, 168.3, 84,06, 79.06 and 39.83 ppm. (in DMSO-d6) Mass spectrum : m/z 191(M+l) (2), 173 (1), 162 (6), 145 (35), 127 (10). 116 (48), 99 (70), 88 (100), 60 (40) and 55 (20). Funetionaiised furanones (Scheme I) are important building blocks in organic synthesis. These lactones are useful for the preparation of optically active ligands and for the synthesis of biologically active natural products. Garcinia acid (la) isolated by the present method was effectively used for the preparation of Ib, Ic, Id, II, III, IV and V (Scheme I). These compounds were synthesized by the reaction of title compound (la), — with methanol and acid (Ib), — with ethanol and acid (Ic), — with benzylalcohol and acid (Id), — with oleium (II), — with BH3-THF (III), — with HBr followed by elimination (IV) — with HBr in methanoi (V) The process of the invention is illustrated by the following example which should not be construed to limit the .scope of the present invention. Example : Dried rinds of the fruits of Garcinia cambogia (2.0 Kg) were cut into small pieces and were soaked in boiling water ibr 20 hours. Extract was collected and the process was repeated 4-5 times. The combined extracts were concentrated to get a thick mass and methanol was added. Pectin was filtered off and the filtrate was concentrated to a syrup. Recovered methanol was preserved use at a later stage. The syrup was made alkaline by adding sufficient quantity of aqueous sodium hydroxide solution at about 80° C and recovered tnethanol was added till two layers separated. The lower layer which contained the sodium salt of Garcinia acid was washed several times with about 50% aqueous mcdianoJ, prepared from recovered methanol The pure sodium sa!f was neutralised with hydrochloric acid was concentrated. Acetone was added and inorganics were filtered off. The filtrate on concentration yielded crude crystals of Garcinia acid, the crude lactone was recrystallised from acetone-chloroform mixture to give pure crystals of the compound (la). Melting point: 178° C Yield : 135.0 g The specific rotation was recorded on a Roudolf polarimeter. [α]D20 : 102.151 deg. We Claim A process for the isolation of Garcinte odd from the fresh or dried rinds of the fruits of Gandnia indica, Gardnia cambogia, Garcfnia atravfridis, comprising the steps of, in sequence : a) subjecting the rinds to extraction to form an extract b) Adding a solvent to the extract to remove pectin and form a filtrate c) Converting the filtrate to form an alkali salt d) Neutralizing the alkali salt with an acid, followed by evaporation, to form a concentrate e) Purifying said concentrate using a solvent to remove inorganic matter as impurtions, to form a second filtrate f) Concentrating the second filtrate to yeld crude Gsrcinia acid g) Rccrystallizing the crude to form pure crystals of Gradniaatid. Z The process for the isolation of Gnrcinia add as doimed in claim 1, wherein step (a) is repeated a plurallty of times, each with a new batch of rinds, to provide a plurality of extract batches, said batches being combined and conoantratad to form a thick syrup. 3 The process for the Isolation of Ganonia add as claimed In claim 1, wherein step (a), the rinds ane cut into small pieces ond soaked in boiling water for 10-20 hours to effect the extraction. 4. The process for the isolation of Garcinia add as claimed In claim 1, wherein step (a), the extraction comprises a soxheit extraction with a solvent chosen from the group consisting of methanol,, ethanol and butanol. 5. The process for the isolation of Gardnia acid as claimed in claim 1, wherein the water extract is prepared by autodaving the rinds, 6. The process for the isolation of Garcinia acid as daimed in claim 1, wherein step (b), the solvent is chosen from a group consisting of methanol, ethanol and butanol. 7. The process for the isolation of Garcinia acid as daimed in claim 5, wherein step (c), comprises concentrating the filtrate, adding an alkali to the filtrate so that the filtrate becomes alkaline, adding an alocohol to the alkaline filtrate to form a upper and lower layer, separating said lower layer and washing with an aqueous alcohol and form an alkali salt paste. a The process for the isolation of Garcinia acid as claimed in claim 1, wherein step (e), the concentrate is triturated with a solvent chosen from the group consisting of acetone, ether and methanol. 9. The process for the isolation of Garcinia acid as claimed in claim 1, wherein (g), a solvent is used to effect recrystallization, the solvent being chosen from the group consisting of acetone, methanol, ethanol, chloroform and dichloroethane. 1O. The process for the isolation of Gancinia acid as claimed in claim 1, as substantially described in complete.

Documents:

2248-del-1998-abstract.pdf

2248-del-1998-claims-(as files).pdf

2248-del-1998-claims-(cancelled).pdf

2248-DEL-1998-Claims.pdf

2248-del-1998-complete specification (as filed).pdf

2248-del-1998-complete specification (granted).pdf

2248-del-1998-correspondence-others.pdf

2248-del-1998-correspondence-po.pdf

2248-del-1998-description (complete)-(as files).pdf

2248-DEL-1998-Description (Complete).pdf

2248-del-1998-drawings.pdf

2248-del-1998-form-1.pdf

2248-del-1998-form-2-(as files).pdf

2248-DEL-1998-Form-2.pdf

2248-del-1998-form-3.pdf

2248-del-1998-form-9.pdf

2248-del-1998-pa.pdf

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