Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Using bacteria
Reexamination Certificate
1998-05-05
2001-06-05
Witz, Jean C. (Department: 1651)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Using bacteria
C435S261000
Reexamination Certificate
active
06242229
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Technical Field of the Invention
The present invention relates to cosmeticlpharmnaceutical compositions comprising, as an active principle therein, an effective amount of culture medium for at least one non-photosynthetic filamentous bacterium, said medium being clarified and stabilized.
The present invention also relates to cosmetic treatments comprising administration of the aforesaid compositions.
2. Description of the Prior Art
It is known to this art to employ microorganisms in cosmetics or in pharmacy, principally in the form of extracts, for their art-recognized properties.
By the term “extract” is intended a product corresponding to a microorganism biomass which, after culturing and once separated from its culture medium, is subjected to various treatments which can range from simple freezing of the biomass to very elaborate purifications of microorganism constituents.
Exemplary are yeast extracts used in cosmetics, bacteria used for the preparation of milk products, bacterial extracts used as bactericides, as cicatrizing agents or, alternatively, as immunostimulants.
More particularly representative is the formulation, into cosmetics, of ribosomal fractions, for example in the preparation of compositions intended to retard aging of the skin by stimulating cell growth and modifying the maturation of connective tissue (EP-A-631,773).
In medicine, compositions for reinforcing the immune defenses are known, in particular for patients who have suffered serious burns and are consequently sensitive to the opportunist infections caused by bacteria, viruses or fungi (WO-91/11174) or, alternatively, as regards diseases of the otorhinolaryngology sphere (FR-2,253,499, FR-2,360,314, FR-2,388,563, FR-2,674,755 and ZA-8801071) or allergies (U.S. Pat. No. 4,946,945).
These compositions generally comprise preparations of microorganism ribosomal fractions.
This is a class of bacteria for which many properties have been demonstrated and which are useful both in cosmetics and in pharmacy. They are the class of filamentous, non-photosynthetic bacteria as defined according to the classification in Bergey's
Manual of Systematic Bacteriology
(Vol. 3, Sections 22 and 23, 9th edition, 1989).
These bacteria, several of which have already been described, generally exist in an aquatic habitat and can be found, in particular, in marine waters or in thermal springs.
These bacteria, or extracts thereof, are described in FR-2,283,223 for their anti-inflammatory and cicatrizing properties, their antiacne and antiseborrhoeic power and their ability to promote moisturization of the skin.
EP-A-0,681,831 describes the formulation of these bacteria or of extracts thereof into cosmetic compositions intended to combat aging of the skin.
The reason for this is that such bacteria or extracts thereof exhibit properties in the field of epidermal renewal.
EP-A-0,761,204 describes these bacteria or extracts thereof for the soothing of sensitive skin, this property moreover being attributed to their substance P-antagonist property.
In WO-94/02158 and in EP-A-0,765,667 the immunostimulatory properties of these bacteria or extracts thereof are described for applications in cosmetics and in pharmacy.
In all of these applications, the active agent is the bacterial biomass, separated from its culture medium and optionally having been subjected to various treatments.
The production of this biomass requires specific growth conditions which make it delicate and increases the time required for the preparation, as well as the cost of this starting material.
SUMMARY OF THE INVENTION
It has now surprisingly and unexpectedly been determined that the culture medium of non-photosynthetic filamentous bacteria, which has been clarified and stabilized, to date considered as being of no value at the end of the culturing and removed, indeed exhibits properties similar to those of the biomass.
Furthermore, even more unexpectedly, the level of expression of these properties by the medium is completely comparable to that of the biomass.
The advantage presented by such phenomenon is immediately apparent, in particular because it is known that culturing, in a 300 liter fermenter, only produces between 270 grams and 360 grams maximum dry weight of biomass.
Thus, the present invention features cosmetic/pharmaceutical compositions comprising, in a cosmetically and/or pharmaceutically acceptable vehicle, diluent or carrier, an effective amount of a culture medium for at least one non-photosynthetic filamentous bacterium, said medium being clarified and stabilized.
DETAILED DESCRIPTION OF BEST MODE AND SPECIFIC/PREFERRED EMBODIMENTS OF THE INVENTION
More particularly according to the present invention, by the expression “cosmetically and/or pharmaceutically acceptable vehicle, diluent or carrier” is intended any physiologically acceptable support substrate.
By the expression “clarified culture medium” is intended a culture medium which has been employed to amplify a microorganism, in particular a bacterium, which, after amplification of said microorganism, has been subjected to a manipulation entailing physically separating it from said culture medium and from said microorganism.
By the expression “stabilized culture medium” is intended a culture medium which has been subjected to a manipulation for preserving it in the state in which it was found at a selected given instant, while at the same time conserving its intrinsic properties. In particular, when this is a microorganism culture medium, this manipulation is intended, for example, to render the medium sterile, i.e., incapable of permitting the growth of microorganisms while at the same time preserving, for example, any biological properties it may have.
Exemplary non-photosynthetic filamentous bacteria according to the invention are preferably those of the order of Beggiatoales. Even more particularly preferred bacteria are those of the genera Beggiatoa, Vitreoscilla, Flexithrix or Leucothrix.
Exemplary such bacteria include:
Vitreoscilla filiformis
(ATCC 15551)
Vitreoscilla beggiatoides
(ATCC 43181)
Beggiatoa alba
(ATCC 33555)
Flexithrix dorotheae
(ATCC 23163)
Leucothrix mucor
(ATCC 25107)
Sphaerotilus natans
(ATCC 13338)
A strain of
Vitreoscilla filiformis
is preferably employed according to the invention.
In order to prepare useful culture media according to the invention, the bacteria can be cultured according to techniques well known to this art. In this regard, all details required for culturing these bacteria are set forth in Bergey's
Manual of Systematic Bacteriology
(9th edition, 1989).
It is known to the art that non-photosynthetic filamentous bacteria are relatively difficult to culture, and the production of pure cultures is likewise difficult. Most researchers recommend the use of poorly defined media, including various macerations using tap water. The carbon source recommended is an acetate.
Preferably, the culture medium according to the invention is the culture medium which allowed the preparation of a biomass of non-photosynthetic filamentous bacteria per the process described in WO-94/02158, assigned to the assignee hereof.
It has thus been shown that it is possible to adapt these bacteria, by counter-selection, to the use of an ose, instead of acetate, as a carbon source.
It has also been shown that it is possible to culture these bacteria on a fully defined culture medium. Culturing can be carried out, in particular, in the following medium:
COMPOSITION
CONCENTRATION
Autolytic yeast extract
0.5 to 5 g/l
Peptone
0.5 to 5 g/l
Anhydrous glucose
0.5 to 7 g/l
Heller microelements
0.5 to 5 ml/
CaCl
2
.10H
2
O
0.01 to 0.20 g/l
The mixture is diluted to 1000 ml with distilled water. Exemplary peptones include soybean papain peptone.
This specific medium differs from the media generally employed by the absence of catalase and of sulfide, although these ingredients were generally hitherto considered as essential.
The Heller microelements, the composition of which is set forth below, were described by Heller,
A
Aubert Lucien
Breton Lionel
Martin Richard
Pineau Nathalie
Burns Doane Swecker & Mathis L.L.P.
Societe L'Oreal S.A.
Witz Jean C.
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