Process for extracting polyphenol fractions of tea and compositi

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Conjugate or complex

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424456, 424474, 424439, 426431, 426435, 426425, 426427, 426428, 426489, 426492, 426478, 426481, 426541, 426542, A61K 3578, A23F 316, A23F 322

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active

059895578

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

The present invention relates to novel polyphenol fractions of Camellia sinensis (tea), the process for the preparation and use thereof, as well as to formulations containing such fractions.


BACKGROUND OF THE INVENTION

More particularly, the invention relates to a process for preparing substantially caffeine-free extracts containing polyphenols derived from epigallocatechin in their natural ratio.
Caffeine is known to have undesirable effects on the cardiovascular system as well as a mutagenic effect, and it is usually removed from such polyphenol fractions by extraction with carbon dioxide under hypercritical condition or with chlorinated solvents. However, the above techniques are not intended to, nor are they capable of, producing extracts with reproducible amounts of the polyphenyl compounds responsible for benificial biological effects.
The polyphenol, characteristic of the tea plant, have recently been the object of a great interest due to their strong antioxidant effect, as described in, for example, Zhao B. et. al., Cell Biophys. 14, 175 (1989) and Huang M. T. et al., Carcinogenesis 13, 947 (1992). Such an effect is apparently related to some extent to the capability of some of these polyphenols (for example epigallocatechin-3-O-gallate) of inhibiting formation of neoplasias artificially induced in the laboratory animals Conney A. H. et al., Prev. Med. 21, 361 (1992) and Mukhtar H. et al., J. Invest. Dermatol. 102, 3 (1994).
Moreover, some recent epidemiological studies evidenced how the consumption of large amounts of green tea is associated with a lower incidence of degenerative chronic diseases, for example some forms of cancer and atherosclerosis see, e.g., Kono, S. et al., Jpn. J. Cancer Res. 79, 1067 (1988); Oguni I. et al., Agric. Biol. Chem. 52, 1879 (1988); Kato I et al., Jpn. J. Cancer Res. 81, 1101 (1990) and Kono S. et al., Prev. Med. 21, 526 (1992)
At present, the mechanisms of action of the substances contained in green tea derivatives have not yet been completely elucidated, apart from the general antioxidant effect described above. It has however, been determined that the polyphenol component of green tea is capable of exerting a differentiated cytotoxic effect depending on the type of cell which is studied and that such a differentiation is particularly related to the selective targeting which some of these polyphenol molecules have towards sub-cell sites such as mitochondria.
Therefore, there has been a long-felt need, which is now met by the present invention, for a method of extracting Camellia sinensis that produces extracts containing a predetermined, i.e., "standardized" amount of polyphenols and wherein the caffeine content of these extracts is minimized, if not eliminated totally.


SUMMARY OF THE INVENTION

The invention therefore is directed, in a first embodiment, to a process for extracting polyphenol fractions from Camellia sinensis, which fractions contain no more than about 0.2% by weight of caffeine. The extraction process comprises the steps of: providing a vegetable biomass of Camellia sinensis containing a quantity of polyphenols; extracting the vegetable biomass with a hydrated solvent, e.g., a water-alcohol or a water-ketone mixture; at least partially evaporating the solvent to form a residue; adding to the residue thus produced an aqueous mixture of from about 30-60% by volume of methanol to form a suspension of the residue; extracting the hydromethanol suspension with at least one chlorinated solvent, e.g., methylene chloride; filtering and concentrating the hydromethanol phase thus obtained; extracting the concentrated hydromethanol phase with at least one solvent from the group including aliphatic esters, alcohols and ketones, with the proviso that the solvent is substantially immiscible with the hydromethanol phase; concentrating the solvent phase from the immediately preceding extraction step; diluting the concentrated solvent phase with at least one chlorinated solvent; and treating the diluted solvent phase with at least one sulfoni

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