Bro-Ooronet,H High Content of Essential Fatty Acids, Which Consist Prac Where N Is = 15 Or 17 and N=17

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Bro-Ooronet,H High Content of Essential Fatty Acids, Which Consist Prac Where N Is = 15 Or 17 and N=17 3,031,478 United States Patent Office Patented Apr. 24, 1962 1. 2 known as arachidonic acid, whose empirical formula is PROCESS FOR THE PRöUCTION3,031,478 OF NATURAL C20H2O2 and whose chemical name is 5,8,11,14-eicosate PHOSPHOLPDS AND SUBSTANCES PRODUCED traenoic acid. THEREBY The essential fatty acid possess a considerable degree Ernst Kenk, 3 Goebenstrasse, Hildegard Debuch, 82 Sie 5 of importance for the human organism. In the numerous bengebirgsalilee, Hans Eikermann, 60 Wiethasestrasse, studies and investigations conducted by medical re and Gertrud Lappe, née Reuter, 113 Bachemerstrasse, Searchers, the following facts have been uncovered: all of Koln, Germany In numerous animal experiments it had been found No Drawing. Filed May 5, 1960, Ser. No. 26,954 that during a non-fatty diet a small amount of linoleic 6 Claims. (CI. 260-403) O acid is sufficient to synthesize arachidonic acid in the The present invention refers to a process for the pro body. duction of natural phospholipids obtained from vegetable In acute parenchyma it has been found that the length crude phosphatides, particularly such which are produced of sickness is shortened considerably by feeding essential in a known manner out of such sources as soya beans, fatty acids. Also in a chronic heptatopathic condition, a and to substances produced thereby. distinct increased degree of success was evident with the The present application is a continuation-in-part of our Same treatment. Essential unsaturated fatty acids, how ever, have also shown not only a lipotropic action, but co-pending patent application, Ser. No. 694,524, filed a favourable influence upon glycogen synthesis as well. November 5, 1957. It is an object, therefore, of the present invention to The choline and colamine phosphoric acid diglyceride produce natural phospholipids rich in essential fatty acids esters belong to crude ester phosphatides which are widely 20 from crude phosphatides of vegetable origin, especially distributed throughout the animal and plant kingdoms. from soya beans containing crude phosphatides. These esters appear as compounds, particularly in the It is another object of the present invention to produce crude phosphatides of vegetable origin. A mixture of from crude phosphatides of vegetable origin, especially crude phosphatides is designated generally in trade and from soya beans containing crude phosphatides, phos industry as lecithin, 25 pholipids wherein choline esters of diglyceride phosphoric However, while according to chemical nomenclature acid predominate, or which consist entirely thereof, and all substances containing choline are classified as lecithins, which are rich in unsaturated and essential fatty acids. those which are characterized by a weaker basic group A further object of the present invention is to produce of B-amino-ethyl alcohol are generally known as cephalins. a Substantially chemically pure choline ester of digly Cephalin (colamine phosphoric acid diglyceride ester) 30 ceride phosphoric acid from crude phosphatides of vege is identified by the following structural formula: table origin especially from soya beans containing crude CO-COCEInt phosphatides in such manner, that the essential fatty acids contained in a natural condition in the phosphatide esters are retained, while other substances, including all phos O 35 phatide esters which do not have choline as their basic component, are removed to a great extent. Another object is to produce a phospholipid with a bro-ooronet,H high content of essential fatty acids, which consist prac where n is = 15 or 17 and n=17. tically exclusively of choline esters of diglyceride phos Lecithin (choline phosphoric acid diglyceride ester) 40 phoric acid and may be designated as chemically pure is identified by the following structural formula: and suitable to be intravenously injected. g-Ho-c CnEI2n+1 Yet a further object of the present invention is to provide a novel product which is obtained in accordance CHO,-COCElem with the novel methods herein described and which is of O 45 therapeutic and medical value. It is an additional object to provide a novel product dro- CCHN(CH3)3OH which may be intravenously injected and which is pro duced in accordance with the novel process hereinde where n is = 15 or 17 and n=17. scribed. On hydrolysis lecithin yields fatty acids, glycerol, phos 50 Other objects of the present invention will become phoric acid and choline. There are, therefore, different apparent during the course of the following specification. lecithins depending on the character of the fatty acid In the attainment of the aforesaid objective, the inven radicals. Each lecithin contains unsaturated and saturated tive concept of the present invention may be realized fatty acids. The saturated fatty acids were found to be through a method of producing a chemically pure choline e.g. palmitic and stearic, and the unsaturated fatty acids 55 ester of diglyceride phosphoric acid exclusively from consist as we have found, to a great extent of so called crude phosphatides of vegetable origin, especially from essential fatty acids. Soya beans containing crude phosphatides which have a It is understood that in medical science at present the high content of essential fatty acids. The inventive con essential fatty acids are considered to be generally those 60 cept of the present invention further includes the feature fatty acids important to life and which the human body that choline phosphoric acid diglyceride esters produced can not itself synthesize. These essential fatty acids are according to the method of this invention may be split of a specific configuration, of which linoleic acid is of off enzymatically with snake venom which attacks and basic significance in the human organism, since it pro splits off fatty acids in their alpha position, so that mono duces in the human body a very active esential fatty acid 65 glyceride phosphoric acid choline esters (lysolecithin) 3,081,478 3 4. are produced, which in the beta position contain a sur preferably aluminum and/or magnesium and/or silicon. prisingly high content of essential fatty acids. Mono For the purpose of the present invention, aluminum oxide glyceride phosphoric acid choline esters are highly effi has been found to be especially useful. Upon evapora cient biologically through the provision of an unusually tion of the alcoholic eluate by distillation under vacuum large external surface activity. They have been found at a temperature of at most 35° C. in the presence of to be capable of being used successfully both physiologi an inert gas, a chemically pure choline ester of diglycer cally for assimilation purposes and medically as heart ide phosphoric acid is obtained. This product has been stimulants. The choline ester of the monoglyceride phos found to be particularly suitable for intravenous admin phoric acid is water soluble as distinguished from the istration and contains a surprisingly high content of un choline ester of diglyceride phosphoric acid, but is insolu O saturated and essential fatty acids. ble in ether and acetone. In order to increase the output of choline phosphoric It is known that the ester of diglyceride phosphoric acid diglyceride esters, the compounds formed in the acid may be extracted from crude phosphatides, particu column from the respective oxides and/or carbonates of larly from soya beans containing crude phosphatides, the aforenamed elements and choline and colamine phos firstly by extracting crude phosphatides by means of ace 15 phoric acid diglyceride esters, must be split off by elut tone and then processing with alcohol the ester mixture ing with alcohol which takes place at a temperature of resulting from this extraction. at most 35 C. in the presence of an inert gas. The The objects of the present invention are realized, how eluate leaving the column and obtained by eluting with ever, by Subjecting an extraction residues of vegetable alcohol may be combined with the first eluate fraction crude phosphatides, such as crude soya bean phospha 20 and both solutions simultaneously concentrated under tides, which has been freed from acetone in vacuum and vacuum again at a temperature of at most 35 C. in the in the presence of an inert gas, to a repeated extrac presence of an inert gas, such as nitrogen. tion with alcohol; the alcoholic solutions are allowed All steps, the extraction, the eluation and the evapora to stand for several days, are then freed from suspended tion are preferably carried out under the exclusion of light. or colloidal particles, which had been also dissolved, 25 The resulting choline ester of diglyceride phosphoric and are finally concentrated in vacuum while passing an acid is a colorless substance of paste-like consistency inert gas. In this manner, phospholipids are obtained which is easily soluble in ether, chloroform, alcohol, in satisfying amounts, which contain choline predomi petroleum ether and other organic solvents. In acetone nantly as the basic component and which have substan it is completely insoluble. The total nitrogen value is tially 60% essential fatty acids in relation to the total 30 theoretically calculated at 2.05%. According to a fatty acid content. Kjeldahl determination, the choline nitrogen value is cal All steps, the extraction with acetone and alcohol are culated at 2.04%; consequently, over 99.5%, or practi carried out at a temperature not exceeding 35 C. and cally 100%, of the total nitrogen consists of choline preferably with the exclusion of light. nitrogen. The fatty acids of the esters - produced in It should be noted in this connection that the alcoholic 35 accordance with the present invention, calculated as total solutions resulting from the extraction are turbid since fatty acids present as ester constituents, are constituted they contain particles which are suspended or colloidally of 17.7% saturated and 82.3% unsaturated fatty acids. dissolved therein. These alcoholic solutions can not be The absolute values solely for the unsaturated fatty acids purified by filtration, since the particles pass through the are as follows: filter.
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