Process for the production of a pyrogen-free fructan which is re

Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives

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4241951, 5361231, C07H 106, C07H 108, A61K 3578

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055612261

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The present invention concerns a process for the production of a pyrogen-free fructan which is readily water-soluble as well as a renal diagnostic agent based on such a fructan that can be applied parenterally.
Fructans, also called polyfructosans, are oligosaccharides and polysaccharides which are constructed of straight or branched fructose chains grafted onto a sucrose parent molecule. Substantial differences can occur in their physical properties depending on the degree of branching and polymerisation such as e.g. solubility in water. Many fructans are stored in plants as reserve carbohydrates and occur predominantly in subterranean parts of Compositae, Campanulaceae, grasses and Liliaceae.
A particular application of fructans is in renal diagnostics especially for the determination of the glomerular filtration rate. Inulin, a fructan from the roots of various species of Compositae such as chicory, dahlias and topinambur, is usually used as the standard test, substance for this (A. N. Richard, B. B. Westfall, P. A. Bott: Proc. Soc. Exp. Biol., N.Y. 32 (1934), 73 and O. Sch uck: Examination of Kidney Function, Martinus Nijnhoff Publishers, Boston 1984, p. 9 ff). Inulin has the advantage that after parenteral application it is neither changed by metabolism nor is it stored in the organism but is filtered by the renal glomeruli and is not reabsorbed in the tubuli (J. A. Shannon, H. W. Smith: J. Clin. Inv. 14 (1935), 393). However, for everyday clinical routine inulin has the disadvantage that it is only very sparingly soluble in water and thus aqueous preparations crystallize during storage and have to be dissolved again by prolonged heating before application. However, during this process depending on the duration of heat exposure inulin is attacked hydrolytically and may be degraded to fructose. A further disadvantage is that if the dissolution is incomplete remnants of undissolved inulin particles remain in the preparation which are not easily detectable and can cause severe circulatory complications after injection. Thus, in the Austrian Patent specification 304769 it is proposed that other water-soluble fructans, namely from the storage organs of plants from the Liliaceae, Amaryllidaceae and Gramineae families such as sinistrin and phlein should be used as a substitute for inulin. Like inulin these are composed of ca. 10 to 40 fructose units corresponding to molecular weights of ca. 1600 to 6500.
In the older literature, O. Schmiedeberg: Hoppe Seyler's Z. Physiol. Chem. 3 (1979), 112 and G. Klein: "Handbuch der Pflanzenanalyse", Springer 1932, 1st part, p 866-873, these fructans are obtained from aqueous extracts of suitable plant organs, after purification with precipitation agents such as e.g. lead salts, by subsequent reprecipitation which is repeated several times with organic solvents such as methanol, ethanol or acetone. The Austrian Patent specification cited above substitutes the use of toxic heavy metal salts by precipitation with the said solvents in an acid environment at pH values of 1.0 to 3.0. In this process partial degradation of the readily hydrolyzable fructan chains, lowering of the average molecular weight and loss in yield can occur. Furthermore, in all these previously known processes extensive safety measures are necessary because of inflammability and danger of explosion and undesired solvent residues may be present in the product.
Thus, it is the object of the present invention to provide a process for the isolation of fructans in which no organic solvents or toxic heavy metals have to be used and in which an improvement in yield is obtained. Moreover, the process should also ensure that pyrogens are separated so that contaminated and infected raw material can also be used.
This object is achieved according to the present invention by a process for the production of a readily water-soluble and pyrogen-free fructan which is characterized in that an aqueous extract, which is obtained in a well-known way from parts of plants containing fructan, is subjected to ultra

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Estelberger et al. Eur. J. Clin. Chem. Clin Biochem., vol. 33(4), 201-209, (1995) [Abstract Only].
Nitsch et al. Carbohydrate Research, vol. 72, pp. 1-12, (1979). [Abstract Only].

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