MXPA00006290A - Use of polyphenol compounds or their derivatives as scavengers of free radicals in cigarette filters - Google Patents

Abstract

The invention concerns the use of polyphenol compounds or their derivatives, obtainable by chemical, biotechnical process or from plant extracts such as rosemary, as scavengers of free radicals in cigarette filters.

Description

USE OF POLYPHENOLIC COMPOUNDS OR THEIR DERIVATIVES AS FREE RADICAL FILTERS IN FILTERS OF CIGARETTE Description of the invention The present invention relates to the use of polyphenolic compounds or their derivatives as free radical scavengers in cigarette filters. Many polyphenolic compounds are known for their beneficial properties in domains as varied as hypertension, hypercholesterolemia, involved in cardiovascular diseases, viral infections, or even inflammatory phenomena. The anti-lipoperoxidants and anticarcinogenic activities of certain polyphenols have also been described. On the other hand, the incorporation of green tea polyphenols in a cigarette filter has been considered to eliminate the unpleasant smell of cigarettes. The authors of the present invention have now discovered that the incorporation of polyphenols or their derivatives into the filter of a REF.121334 The cigarette allows efficient elimination of the free radicals of the cytotoxic molecules of cigarette smoke at the moment of its passage through the filter. The subject of the present invention is the use of polyphenolic compounds as free radical scavengers ("scavengers") in cigarette filters. Preferably, the polyphenols used according to the present invention can be chosen from carnosol, rosol anol, rosin aric acid, carnosic acid, and their derivatives. "Derivatives" are understood primarily as compounds that are derived from compounds polyphenols by substitution of the hydrogen atom of at least one of the hydroxyl groups of the polyphenolic compounds with an alkyl group of _ 1 to 6 carbon atoms, or a group (alkyl of 1 to 4 carbon atoms) carbonyl. Acetates such As the carnosic acid acetates and the rosmarinic acid acetates are preferred derivatives of the polyphenolic compounds used according to the invention. By "derivatives" of the polyphenols used according to the invention such as carnosol, rosmanol, rosmarinic acid, carnosic acid, are also understood the isomers of said polyphenols mainly such as epirrosmanol and isorrosmanol (Nakatani et al. Agrie. Biol. Chem. 1984, vol 48, No. 8, pp 2081-2085). These compounds can be obtained by classical chemical synthesis or by biotechnological route, according to the methods known to the person skilled in the art. These might be. equally isolated from plant extracts. Said polyphenols can also be used according to the present invention in the form of a plant extract, preferably of a plant extract of the Labiada family, mainly such as a rosemary extract Rosmarinus officinali s L). A plant extract of this type can be obtained by extraction with a polar solvent such as an alcoholic or hydroalcoholic solvent. The alcohol used as a solvent can be mainly ethanol. This extract can also be advantageously obtained with the help of supercritical carbon dioxide and is then richer in polyphenolic compounds. Preferably, the plant extract used according to the invention can be obtained by extraction with a polar solvent followed by an extraction with supercritical C02. Rosemary extraction is preferably carried out on dry plants, for example on branches of rosemary, cut and dried in the sun for 4 to 5 days. The polyphenolic compounds or their derivatives, obtained by chemical synthesis, by biotechnological route, or by extraction from vegetables, can be used alone or in a mixture according to the invention. Preferably, a mixture of carnosol, carnosic acid and rosmarinic acid can be sold. The polyphenolic compounds or their derivatives can be used under the free form or they can be conjugated or coupled to a support ("carrier") that allows to increase the weight of the polyphenolic set that can be constituted, for example, of plastic (polystyrene or others) or of glass.

The polyphenolic compounds can be incorporated in the cigarette filters at a ratio of 0.5 mg to 0.5 grams, preferably 0.002 to 0.1 gram, preferably 0.01 grams. When the polyphenolic compounds are dissolved in a solvent, it is advantageous to embed the filter of the cigarette with said solvent containing the polyphenols, and then proceed to the evaporation of the solvent. More particularly, the filter of the cigarette can be soaked or soaked with an alcoholic or hydroalcoholic vegetable extract and then subjected to an evaporation of the alcoholic or hydroalcoholic solvent. The polyphenolic compounds or their derivatives can also be dissolved in a saturated oil, and thus incorporated in the cigarette filter. Another object of the invention is a process for the preparation of a cigarette filter in which the polyphenolic compounds chosen from carnosol, rosmanol, rosmarinic acid, carnosic acid and their derivatives are incorporated into said cigarette filter. The invention also has as its objective a cigarette filter obtained by the process *! previously described. Such a filter allows to reduce the amount of molecules with free radicals present in the cigarette smoke. The authors of the present invention have discovered that the polyphenols or their derivatives incorporated in the filter of a cigarette capture the free radicals of the cytotoxic molecules present in cigarette smoke, both in the aqueous phase and in the solid phase of the smoke, which It consists essentially of tars. These cytotoxic molecules favor the appearance of cancers in smokers, mainly lung cancer. Polyphenols or their derivatives incorporated in the filter of a cigarette also have, on the one hand, an inhibition of the activity of the carcinogenic compounds, by reducing the formation of heterocyclic amines, at the time of combustion of the tobacco, and, on the other hand, part, a detoxification of carcinogenic compounds, such as benzopyrene. Figure 1 and the following examples illustrate the invention without limiting its scope. The attached figure 1 is a graph representing the intensity of the signal, in electronic paramagnetic resonance, of the terbutyloxy radicals in the gaseous phase of cigarette smoke, as a function of the amount of rosemary extract incorporated in the filter.

EXAMPLE A. Preparation of a rosemary extract. Rosemary ears (Rosmarinus offi cinali s L.) are subjected to an extraction with ethanol at 65 ° C. The volume of ethanol used (in liters) corresponds to five times the weight in kg of the rosemary spikes. The extract is then purified and enriched in polyphenols by selective extraction with supercritical C02. According to the regulation of the temperature between 40 ° C and 100 ° C and the pressure between 1 and 170 bar, the extract is purified and enriched selectively in its different components. An extract of this type contains the following compounds: carnosol, - rosmanol - rosmadial, carnosic acid genk uanina, rosmarinic acid ... The proportions of these different compounds vary according to the rosemary plant used. In general, an extract comprising about 25% rosmarinic acid, about 10% carnosic acid and about 5% carnosol is obtained.

B. Incorporation of the extract in a cigarette filter. A cigarette filter is imbibed either with the prepared extract, the ethanol which is then evaporated, or an oily phase based on CRODAMOL® (Francaise d'Aromes et Parfums), in which a powder obtained by drying is dissolved. extract prepared previously.

C. Efficacy of the filter that contains the polyphenols. 1. Computer aided modeling: The effectiveness of the cigarette filter prepared in this way is evidenced in a first time by computer-aided modeling, according to the Monte-Cario method, which allows calculating the number of encounters between a carcinogenic target molecule and a polyphenolic compound used according to the invention. The number of cytotoxic molecules with free radicals present in cigarette smoke has been calculated on the one hand and the other on the filter. The number of cytotoxic molecules is a function of the volume of smoke that passes through the filter, the volume of the filter, the concentration of cytotoxic molecules in the smoke and the concentration of polyphenols in the filter. The authors of the present invention have thus shown that 0.01 g of the rosemary extract incorporated in the filter of a cigarette makes it possible to reduce by more than 70% the rate or proportion of molecules with cytotoxic free radicals in cigarette smoke. 2. Electronic paramagnetic resonance The antiradical activity of the extract comprising carnosol, rosmarinic acid and Carnosic acid has been confirmed by an Electronic Paramagnetic Resonance Spectroscopy (RPE) that eventually puts into operation the so-called "spin trapping" method. to. Materials and methods . The RPE is a technique that allows to study directly or indirectly the paramagnetic substances. Paramagnetic compounds are molecules that have a free unpaired electron on their last valence shell. Such substances are designated with the term radicals. When these radicals have very short lifetimes (much less than a second) it is advantageous to use the so-called "spin" technique. This consists of using traps capable of stabilizing the radicals produced by the formation of an adduct and thus measuring them for several minutes. This reaction is carried out in the following way: Radical life span short trap? adduct (paramagnetic) (diamagnetic) (paramagnetic) DMPO (5,5'-dimethyl-pyrrolin-N-oxide) has been used as a "trap".

Anti-radical activity on the radicals produced in the gaseous phase of cigarette smoke The device that allows entrapment of the radicals of the gas phase of cigarette smoke is described in Pryor et al.

Environmental Health Perspectives, 1976, vol. 16, pp 161-175. A cigarette is placed at the end of a device of this type and the gaseous phase of the smoke, after passing through a Cambridge filter (finer than a cigarette filter in order to retain the tars), dissolves in a benzene solution "spin spin trap". For each measurement, the DMPO is adjusted to a concentration of 32 mM in a benzene solution, and the gaseous phase of two cigarettes is used.

Antiradical activity on the radical semiquinone contained in the solid phase of the cigarette filter: In addition to the free radicals produced in the gas phase of the smoke, free radicals are present in the tars partially stopped by the solid phase of the commercial classical filters. The radical, mainly present in these tars and responsible for a well-established carcinogenic activity nowadays, is of a semi-quinonic nature. This semiquínone that has a long life will be able to react within the cells and also produce harmful species such as oxygenated radicals. A second series of experiments consisted in the study of the radical semiquinone contained in the solid phase of the filter. This study does not need the use of traps. In effect, the detection of the radical that has a very long life, is carried out in itself over the recovered filter. b. Resulted Efficacy in rosemary extract solution on entrapment of free radicals produced in the gas phase. In order to evaluate the capacity of rosemary extract to trap the free radicals of the gas phase of cigarette smoke in benzene solution, the signal intensity in RPE of the terbutyloxy radicals has been evaluated according to the amount of extract of rosemary added in the entrapment solution, expressed in grams of dry extract powder. The decrease in the signal indicates that rosemary extract has a faster entrapment rate than that of DMPO (see figure).

Efficiency of the rosemary extract on the entrapment of the radicals produced in the gas phase, after the impregnation of the filter with the dry rosemary extract and distem CRODAMOL®: The filter was impregnated with 250 μl of the CRODAMOL®-based lipid phase containing 10 mg / ml dry rosemary extract powder. The RPE analysis of the spectrum of the terbutyloxy radical (tBuO °) dissolved in the benzene allowed to observe a 65% decrease in the DMPO / tBuO ° adduct signal. This result shows that the rosemary extract used is effective to inhibit the formation of the tBuO ° radicals.

Efficiency of rosemary extract on entrapment or radicals produced in the solid phase, after impregnation of the filter with dried rosemary extract and dissolved in CRODAMOL®: Parallel to the experiment carried out previously, the signal of the semiquinone radical present in the cigarette filter has been evaluated. A decrease of approximately 63% of the signal of the semiquinone has been observed after the treatment of the filter with the rosemary extract. In addition, it is interesting to note that no new signals appear that may indicate the formation of new radicals due to the entrapment by the rosemary extract. In As a result, these results show that the rosemary extract is active in terms of its ability to extinguish the signal of the semiquinone, and that it does not suffer, at the time of this reaction, transformation that generates a new radical.

It is noted that in relation to this date the best method known to the applicant to carry out the aforementioned invention is that which is clear from the present description of the invention.

Claims (12)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. The use of polyphenolic compounds or their derivatives present in the form of a rosemary extract, as free radical scavengers in cigarette filters.

2. The use of polyphenolic compounds chosen among carnosol, rosol anol, rosmarinic acid, carnosic acid, and their derivatives, as free radical scavengers in cigarette filters.

3. The use according to claim 2, wherein the polyphenolic compounds are constituted by a mixture of carnosol, carnosic acid and rosmarinic acid.

4. The use according to claim 2, wherein the free radical scavengers are in the form of a mixture wholly or partially composed of carnosol.

5. The use according to claim 2, wherein the free radical scavengers are in the form of a mixture wholly or partially composed of carnosic acid.

6. The use according to any of claims 2 or 3, wherein the polyphenolic compounds are present in the form of a plant extract.

7. The use according to claim 4, wherein said plant extract is a rosemary extract.

8. The use according to any of claims 1, 6 or 7, in which the plant extract is obtained by extraction in an alcoholic or hydroalcoholic solvent.

9. The use according to any of the preceding claims, wherein the polyphenolic compounds are coupled to a polymer type support.

10. The use according to any of the preceding claims, wherein the polyphenolic compounds or their derivatives are incorporated in a cigarette filter at a ratio of 0.5 mg to 0.1 gram, preferably 0.01 gram.

11. A method of preparing a cigarette filter, characterized in that polyphenolic compounds as defined in accordance with any of the preceding claims are incorporated into said cigarette filter.

12. The cigarette filter, characterized in that it is obtained by the method according to claim 11.