FR2777205A1 - Production of pollen extract useful as antifungal agent and in cosmetic, dermatological and pharmaceutical compositions - Google Patents

Abstract

Production of a pollen extract comprises contacting pollen with an extractant comprising pressurized carbon dioxide (CO2) and/or a polar or nonpolar solvent and separating the pollen extract from the extractant.

Description

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  The present invention relates to a pollen extraction process, and cosmetic, dermo-cosmetic or

  pharmaceutical extract obtained by this process.

  The use of pollen in cosmetics has been known for a long time, and a process for extracting pollen from the pollen aggregates collected by bees is described in particular in Japanese patent J52-090635. This process is a classic process of alcoholic extraction, simply applied to pollen. This traditional process consists in distilling the mixture constituted by pollen and

  alcohol, and therefore requires a heating and evaporation step.

  These steps inevitably change the composition of the pollen.

  In addition, the pollen grain consists of an envelope

  lignin, which forms a shell with a microscopic hole.

  The active substances, and in particular the sterols, are found inside the shell. The solvents usually used in the prior art for the extraction of pollen very difficult to penetrate into the pollen grain through the microscopic hole, and consequently only allow an incomplete extraction of the lipid part, and therefore of the sterols, lying inside the shell. To overcome these drawbacks in particular, an object of the present invention is to propose a pollen extraction process characterized in that, in a first phase of the extraction, the pollen is brought into contact with a solvent or a mixture of solvent, said solvent or mixture of solvents being carbon dioxide under pressure and / or a polar or non-polar solvent, then in a second phase, a means is used to separate the pollen extract from the mixture obtained at the outcome of

the first extraction phase.

  According to a preferred embodiment of the invention, the carbon dioxide is at a temperature between 31 C and 70 C and at a

  pressure between 73.10-5 Pa and 300.10-5 Pa.

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  Advantageously, the mixture of solvent used in the first extraction phase is a mixture of carbon dioxide and a

  polar solvent. Preferably, this polar solvent is ethanol.

  According to a particular embodiment of the invention, a treatment of the raw material is carried out, prior to extraction. Preferably, this prior treatment of the material

  first is to grind the clumps or balls of pollen grains.

  1 0 According to a preferred embodiment of the invention, the means of

  separation is to lower the pressure of carbon dioxide.

  Advantageously, the pressure of the CO2 is lowered by carrying out

  several levels, preferably two levels.

  According to another preferred embodiment of the method according to the invention, at the end of said second phase, the CO2 is recycled by lowering its temperature between 0 and 10 C and its pressure to 50.10-5 Pa, then raising the pressure to 300.10-5 Pa by means of a pump and finally by heating it to the desired temperature for

extraction.

  Another object of the invention is to provide an antifungal agent, characterized in that it comprises a pollen extract obtained by the

method according to the invention.

  The object of the invention is also to provide a cosmetic or dermocosmetic composition, characterized in that it comprises a pollen extract obtained by the process according to

  the invention, in combination with any suitable excipient.

  Another object of the invention is to provide a pharmaceutical composition, characterized in that it comprises a pollen extract obtained by the process according to the invention, in combination with

  any pharmaceutically acceptable excipient.

  Advantageously, the cosmetic or dermo-cosmetic or pharmaceutical composition according to the invention is in the form of a gel,

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  cream, lotion, stick or any other vector intended for topical application, or in the form of tablets or capsules or any other

  form intended for oral administration.

  The invention also relates to the applications of a pollen extract obtained by the process described above for obtaining cosmetic compositions intended to combat skin aging, to prevent the harmful effects of solar aggressions, to soothe inflammations due to solar aggressions and to regenerate the cutaneous tissues which have undergone such aggressions and to

  restructure or firm the skin tissue.

  Yet another object of the invention is the application of the pharmaceutical composition according to the invention to obtain a medicament intended for the treatment of acne, psoriasis, burns, varicose ulcers, scars, rheumatism, l asthenia and in particular male sexual asthenia,

  prostatic hypertrophy, or hyperlipidemia.

  Finally, the object of the invention is also to provide a medicament characterized in that it comprises a pharmaceutical composition

according to the invention.

  The invention will be better understood on reading the description

  detailed below which illustrates the invention without limitation.

  The method according to the invention is a pollen extraction method characterized in that, in a first phase of the extraction,

  puts the pollen in contact with a solvent or a mixture of solvent.

  Of course, this process does not involve any distillation step: the active components of the pollen extracts obtained by the process according to the invention therefore have structures and a composition strictly identical to those of the starting pollen grains. The pollen extract obtained by the process according to the invention comprises 20 to 60% by weight of unsaponifiable matter. These

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  unsaponifiable matters in particular include waxes (approximately 14% by weight), hydrocarbons (approximately 10 to 15% by weight), sterols, probably in the majority in the form of esters

  (between 5 and 20% by weight) and aliphatic and terpene alcohols.

  The solvent or mixture of solvents used in the process according to the invention is preferably carbon dioxide under pressure and / or a polar or non-polar solvent. Carbon dioxide can be used as the sole solvent. Carbon dioxide is a

  inert gas, present in the air, and completely harmless.

  The process according to the invention can therefore be carried out so as to avoid any contamination of the raw material, as well as of

  the extract, by more or less toxic solvents.

  In addition, the method according to the invention provides no contamination in the environment and is perfectly natural. In a second phase, a means is used to separate the pollen extract from the mixture obtained at the end of the first extraction phase. Carbon dioxide in supercritical or subcritical state, that is to say when it is brought to a temperature close to its critical temperature, namely 31 C and at a pressure higher than its critical pressure, namely 73.10-5 Pa , behaves like a nonpolar solvent, selective to extract the least polar raw materials of pollen, lipids. Under pressure, supercritical carbon dioxide easily penetrates the pollen grain, and even expands the pollen grain. This expansion phenomenon is observed at each extraction, and its importance is such that it conditions the filling of the extractor: it is considered that the volume of the pollen grains, under the action of dioxide of

  supercritical carbon, increases by about 20%.

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  Thus, the introduction of carbon dioxide into the pollen grain being carried out without difficulty and in significant quantities, all the lipids of interest are capable of being extracted from natural products while the proteins and the carbohydrates are not soluble in carbon dioxide. Furthermore, by modifying the temperature and pressure parameters, the extraction of lipids

is done selectively.

  Preferably, the carbon dioxide is brought to a temperature

  1 0 between 31 C and 70 C and at a pressure between 73.10-

  Pa and 300.10-5 Pa. Advantageously the temperature is between 40 C and 70 C and the carbon dioxide pressure is between 200 and 300.10-5 Pa. Generally, it is observed that the extraction yield increases with the temperature and the

pressure.

  According to a preferred embodiment of the invention, the polarity of the carbon dioxide can be modulated on the one hand in a moderate way by playing on the pressure-temperature parameters, on the other hand in a significant way by adding a polar co-solvent . Preferably, the polar co-solvent is added in

  proportions preferably between 1 and 10% by weight.

  Advantageously, the polar co-solvent is an alcohol, preferably ethanol.

  According to a particular embodiment of the invention, the co-

  polar solvent is introduced into the extractor by an auxiliary pump which sends this co-solvent between the carbon dioxide pump and the reheating exchanger. The co-solvent can be

  Almost entirely recovered with the extracts and recycled.

  Good solvent at high pressure, carbon dioxide loses all its capacity to dissolve organic molecules when it is brought to the gaseous state at a temperature close to its critical temperature

  but at a pressure significantly lower than its critical pressure.

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  Thus, the invention makes it possible to separate the products dissolved in carbon dioxide at high pressure by lowering the pressure without

  significantly change the temperature.

  The carbon dioxide pressure can be lowered abruptly, or by respecting several stages. Preferably, as many steps are carried out as there are groups of molecules to be separated: preferably, two or three steps are carried out. The fact of providing pressure levels makes it possible to fractionate the extracts by classifying the molecules composing them according to their solubility in carbon dioxide at different pressures. The recovery of the extracts obtained in the two or three separation stages is carried out by

an automated airlock system.

  According to a particular embodiment of the invention, a treatment of the raw material is carried out, prior to extraction, which preferably consists in grinding the balls of pollen grains, by any known grinding means. This preliminary stage makes it possible, with certain solvents, to obtain better yields

extraction.

  According to a preferred embodiment of the invention, the carbon dioxide used for the extraction is recycled. The recycling is carried out by liquefying the carbon dioxide which was used for the extraction by lowering the temperature thereof, so as to liquefy this carbon dioxide which has passed to the gaseous state after the separation stages. According to a particular embodiment of the invention, the temperature is lowered between 0 and 10 C and the pressure is reduced to 50.10-5 Pa. Then, to reuse this carbon dioxide, its pressure is raised to a higher pressure to 73.10-5 Pa., preferably to 200 to 300.10-5 Pa. The carbon dioxide is then reheated in an exchanger to a temperature preferably between 40 and 70 C,

  before being introduced into the extractors.

  Carbon dioxide is therefore almost completely recycled.

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  Pollen has a very rich composition; it provides proteins, many amino acids, vitamins, minerals, enzymes and provitamin A. The recognized traditional actions of pollen generally consumed in dried balls, are the following: - regulating action of the intestinal functions, as well cases of diarrhea and constipation, - increased hemoglobin in anemia, increased appetite and weight in emaciated individuals and rapid recovery of strength after influenza, depression and other diseases, - beneficial action on intellectual fatigue, probably thanks to its high content of amino acids, - fortifying action on the circulatory system and in particular capillary, by the presence of rutin, glucoside preventive of hemorrhages and fortifying the contractions of the heart, - positive action on the growth of young children, - action rejuvenating by recovery of male potency, - beneficial action on the functions of the fo ie and rapid recovery from health after jaundice, very effective curative action of prostatitis, - diuretic action,

  - favorable action on lipid metabolism.

  However, the inventors have highlighted unknown and completely new properties for the extracts obtained by the process according to

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  the invention. Thus, it appears that the pollen extract obtained by the process according to the invention has particularly advantageous pharmacological properties. The inventors have in particular demonstrated that the pollen extract increases significantly, at concentrations of 8.10-4%, 4.10-3 % and 2.10-2% by weight, the neosynthesis of total collagens (secreted collagens + collagens

  intracellular) and the multiplication of fibroblasts.

  These pharmacological properties lead to various dermocosmetic applications, mainly in the field of cosmetics or dermo-cosmetics. The multiplication of fibroblasts shows that the pollen extract obtained by the process according to the invention is a restructuring, firming and cytostimulating compound and can therefore be used in particular in cosmetic compositions intended to combat skin aging, for example anti-wrinkle, anti-stretch mark compositions, or aimed at obtaining a smoothing and softening effect on the skin, by

example an anti-dimpling effect.

  Furthermore, the inventors have also demonstrated that the pollen extracts according to the invention, at selected concentrations, allow the resumption of the multiplication of fibroblasts previously weakened by UVA radiation. These pharmacological properties lead to dermocosmetic or pharmaceutical applications as a repair and healing agent. Among the cosmetic applications, the most important are those relating to the protection of the skin, as a preventive measure, against the effects of the sun, and to the soothing and regeneration of a skin.

  previously subjected to solar aggression.

  It is understood that the compositions according to the invention can be in any suitable dosage form, and in combination with any cosmetically or pharmaceutically acceptable excipient, for oral or

topical.

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  The inventors have also demonstrated that the pollen extract obtained by the process according to the invention has an antifungal activity and can be used as an anti-fungal agent, both in cosmetic compositions and in dermocosmetic, pharmaceutical or parapharmaceuticals, especially in the field of podiatry or in

  anti-dandruff compositions.

Example 1:

  Two 400 liter autoclaves connected in series, kg of pollen are introduced into an autoclave, the autoclave is closed and the CO2 is introduced at a pressure of 280.10-5 Pa. The flow rate of the C02 introduced is between 600 and 1800 kg / h. During the entire extraction phase, the CO2 is kept at a temperature between 50 and 60 C. The extraction phase lasts 4-5 hours, but may vary depending on the capacity of the pump. The separation is carried out in a two-stage cyclonic separator, which allows gravity separation to be carried out in the following two stages: in a first stage of the separator, the pressure is lowered from 280 to 60.10-5 Pa, and the temperature is brought back between 30 and 50 C. The total separation of the pollen extract is carried out in the 2nd separation stage, at a pressure between 40 and 50 × 10 -5 Pa and a temperature between 40 and 50 C. The yield is approximately 2.5%. An orange-brown oily paste is obtained, with a strong sweet aromatic odor and a

  high lipid content (60 to 80% by weight).

Example 2:

  2 mg of pollen extract are deposited on an agar medium poured into a Petri dish and seeded on the surface using a culture in

  liquid medium of an Aspergellus niger mold CIP 183183.

  After incubation, the results are read by measuring the zones of inhibition in millimeters, ie for Aspergellus niger: 10 mm. The pollen extract obtained by the process according to the invention

  3 5 therefore reveals an antifungal activity.

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Claims (20)

  1. Pollen extraction method characterized in that, in a first phase of the extraction, the pollen is brought into contact with a solvent or a mixture of solvents, said solvent or mixture of solvents being carbon dioxide under pressure and / or a polar or apolar solvent, then in a second phase, a means is used to separate the pollen extract from the   mixture obtained at the end of the first extraction phase.   2. Method according to claim 1, characterized in that said carbon dioxide is at a temperature between 31 C and 70 C and at a pressure between 73.10-5 Pa and 300.10-5 Pa.   3. Method according to any one of claims 1 or 2,   characterized in that said mixture of solvent used in the first extraction phase is a mixture of dioxide of carbon and a polar solvent.   4. Method according to claim 3, characterized in that said polar solvent is ethanol.   5. Pollen extraction method according to any one of   claims 1 to 4, characterized in that,   prior to extraction, processing of the raw material. 6. Pollen extraction method according to claim 5, 3 0 characterized in that said preliminary treatment of the material   first is to grind the balls of pollen grains.   7. Pollen extraction method according to any one of   claims 1 to 6, characterized in that said means of   separation is to lower the pressure of CO2.   i 1 2777205 8. Pollen extraction method according to any one of   claims 1 to 7, characterized in that the   CO2 pressure by performing several steps.   9. Extraction method according to claim 8, characterized in that   lower the CO2 pressure by performing two steps.   10. Pollen extraction method according to any one of   claims 1 to 9, characterized in that, at the end of said   second phase, the CO2 is recycled by lowering its temperature between 0 and 10 C and its pressure to 50.10-5 Pa, then raising the pressure to 300.105 Pa by means of a pump and finally by   reheating to the desired temperature for extraction.   11. Antifungal agent characterized in that it comprises a pollen extract obtained by the process according to any one of claims 1 to 10.   12. Cosmetic or dermocosmetic composition, characterized in that it comprises a pollen extract obtained by the process   according to any one of claims 1 to 10, in   combination with any suitable excipient.   13. Pharmaceutical composition, characterized in that it comprises a pollen extract obtained by the process according to   any one of claims 1 to 10, in association with   any pharmaceutically acceptable excipient.   14. Cosmetic or dermo-cosmetic composition or   pharmaceutical according to any one of claims 12   or 13, characterized in that it is in the form of a gel, cream, lotion or any other vector intended for topical application, or in the form of tablets or capsules or any   another form for oral administration.   15. Application of a pollen extract obtained by the process according to   any one of claims 1 to 10 for obtaining 1 2 2777205   of a cosmetic composition intended to fight against skin aging.   16. Application of a pollen extract obtained by the process according to   any one of claims 1 to 10 for obtaining   of a cosmetic composition intended to prevent the effects   harmful to the skin from sun damage.   17. Application of a pollen extract obtained by the process according to   any one of claims 1 to 10 for obtaining   of a cosmetic composition intended to soothe the inflammations due to solar aggressions and to regenerate   cutaneous tissues having undergone such attacks.   1 5 18. Application of a pollen extract obtained by the process according to   any one of claims 1 to 10 for obtaining   a cosmetic composition intended to restructure or firm the skin tissue.   19. Application of the pharmaceutical composition according to one   any of claims 13 or 14 for obtaining a   medicine intended for the treatment of acne, psoriasis, burns, varicose ulcers, scars, rheumatism, asthenia (and in particular male sexual asthenia), prostatic hypertrophy, or hyperlipidemia . 20. Medicinal product, characterized in that it comprises a composition   pharmaceutical according to any one of claims 13 or 14.