Process for the preparation of active principles on thermal water and compositions comprising them

Abstract

The invention relates to a process for the preparation of a cosmetic or dermatological active principle, comprising the culturing of at least one nonphotosynthetic and nonfruiting filamentous bacterium on a medium comprising at least one nonsulphurous mineral and/or thermal water. It also relates to a cosmetic or dermatological composition comprising at least one bacterium of the genus Vitreoscilla or one of its extracts.

Claims

1. A process for preparing a cosmetic or dermatological active principle, comprising: culturing at least one nonphotosynthetic and nonfruiting filamentous bacterium comprising an envelope on a medium comprising at least one first water which is thermal water from La Roche-Posay, the first water being mixed with at least one second water which is an osmotically treated and/or distilled water and the concentration of the first water is from 0.5% to 50% with respect to the second water to produce a cultured bacterium comprising an envelope, wherein the at least one first water exhibits a total mineral content of greater than or equal to 400 mg/l; separating the cultured bacterium from the medium, and rupturing the envelope of the cultured bacterium to produce the active principle.

2. The process according to claim 1, wherein a genus of the bacterium is the genus Vitreoscilla.

3. The process according to claim 2, wherein a specie of the bacterium is the specie, Vitreoscilla filiformis.

4. The process according to claim 3, wherein the bacterium is the strain deposited at the ATCC under the reference ATCC15551.

5. The process according to claim 1, wherein a concentration of calcium ions in the first water is greater than or equal to 100 mg/l.

6. The process according to claim 1, wherein a concentration of hydrogen-carbonates in the first water is greater than or equal to 300 mg/l.

7. The process according to claim 1, comprising: fermenting the cultured bacterium in the medium prior to separating the cultured bacterium from the medium to produce a fermented cultured bacterium; and separating the fermented cultured bacterium from the medium to recover a mass of the bacteria; wherein the medium further comprises a monosaccharide, and the bacterium is of the order Beggiatoales.

8. The process according to claim 7, further comprising: stabilizing the mass of the bacteria which is recovered after fermentation.

9. The process according to claim 1, wherein the medium comprises a mixture of (i) osmotically treated or distilled water and of (ii) thermal water, and a ratio (i)/(ii) is between 1 and 100.

10. The process according to claim 1, wherein the at least one first water exhibits a total mineral content of 400 mg/1 to 900 mg/l.

11. The process according to claim 1, wherein the medium comprises a mixture of (i) osmotically treated or distilled water and of (ii) thermal water, and a ratio (i)/(ii) is between 1 and 25.

12. A process for preparing a cosmetic or dermatological active principle, comprising: culturing at least one nonphotosynthetic and nonfruiting filamentous bacterium on a medium comprising at least one first water which is thermal water from La Roche-Posay, the first water being mixed with at least one second water which is an osmotically treated and/or distilled water and the concentration of the first water is from 0.5% to 50% with respect to the second water to produce a cultured bacterium, wherein the at least one first water exhibits a total mineral content of greater than or equal to 400 mg/l; and separating the cultured bacterium from the medium to prepare an extract of the medium, wherein the extract is the active principle.

13. The process according to claim 12, wherein the extract is supernatant from the medium.

14. The process according to claim 1, wherein the concentration of the first water is from 0.5% to 20% with respect to the second water.

15. The process according to claim 1, wherein the concentration of the first water is from 0.5% to 2% with respect to the second water.

16. The process according to claim 12, wherein the concentration of the first water is from 0.5% to 20% with respect to the second water.

17. The process according to claim 12, wherein the concentration of the first water is from 0.5% to 2% with respect to the second water.

Description

EXAMPLE 1

Preparation of the Biomass

Preparation of the Culture Medium

(1) Composition:

(2) TABLE-US-00003 *Yeast extract 2 g *Soybean papain peptone 2 g *Glucose 2 g *Heller microelements 2 ml *CaCl.sub.2•2H.sub.2O 66.21 mg *Thermal water from q.s. for 1 litre La Roche-Posay

(3) The pH of the medium is adjusted to 5.00 with a molar H.sub.3PO.sub.4 solution. The medium is sterilized by autoclaving at 121° C. for 30 minutes. After cooling to ambient temperature, the pH is readjusted to 7.20 by addition of a molar KOH solution.

(4) Culturing:

(5) After inoculating the medium at 1%, the culture is placed under orbital shaking at 100 revolutions/min and at 26° C. After growing for 48 hours, the culture is centrifuged at 8000 g for 15 minutes. The pellets are recovered and then autoclaved at 121° C. for 30 minutes. This biomass can be used for evaluation tests.

EXAMPLE 2

Comparison of the Effect of a Biomass Obtained with or without Thermal Water

(6) The aim of this study is to try to determine the effects on differential gene expression in human keratinocyte cultures of the biomass obtained by culturing on a culture medium reconstituted with osmotically treated water, of the biomass obtained by culturing on a medium reconstituted with thermal water from La Roche-Posay and of thermal water from La Roche-Posay.

Keratinocyte

(7) TABLE-US-00004 Product tested Corneodesmosin LEP16 Biomass — — Biomass LRP +130% +128% Thermal water LRP — —

(8) In a keratinocyte culture, only the biomass cultured on thermal water from La Roche-Posay stimulates the expressions of the corneodesmosin and LEP16. The biomass cultured on osmotically treated water or the thermal water from La Roche-Posay do not have an effect on the expression of these marker genes for keratinocytes. In parallel, the ultrapure water from Merck used as control has no effect on the expression of the genes under consideration.

EXAMPLE 3

Differences in Expression Between the culturing of Vitreoscilla filiformis on Water from La Roche-Posay Versus the Culturing of Vitreoscilla filiformis on Osmotically Treated Water

(9) Two-dimensional electrophoresis was used as means for separating and visualizing the proteins expressed in the Vitreoscilla filiformis biomass, in one case cultured on water from La Roche-Posay and in the other case cultured on osmotically treated water.

(10) This method of analysis consists in separating the proteins as a function of their pHi in a first “horizontal” dimension (this is isoelectric focusing or IEF). This separation takes place via a gel comprising an immobilized pH gradient, also known as IPG strips.

(11) The proteins are subsequently separated as a function of their molecular weights in a second “vertical” migration dimension. The results show that the 2D gels are very rich in proteins; the latter are distributed in a molecular weight range between 6 kDa and 100 kDa, with an abundance of proteins in the region of acidic pH values. The rapid analysis of the image shows 6 different spots, on the one hand, the overexpression of two proteins in the biomass cultured on water from La Roche-Posay and, on the other hand, 4 proteins which appear to be more expressed in the biomass cultured on osmotically treated water.

(12) These results show that there exists a difference in level of expression of the proteins in the biomasses cultured on osmotically treated water in comparison with that cultured on water from La Roche-Posay.

EXAMPLE 4

Increase in the Growth with Water from Vittel

(13) Tests on culturing Vitreoscilla filiformis in a 10-litre fermenter in continuous mode were carried out starting from media partially reconstituted with osmotically treated water (control test) and with mineral water from Vittel (at 2%). The degree of growth set by the continuous culturing method was fixed at 0.12 h.sup.−1 and the culturing conditions were identical in the 2 tests. The biomass was concentrated by centrifuging a continuously harvested culture volume. The measurement of the absorbance at 600 nm makes it possible to monitor the growth of the culture. The sludges obtained were stabilized by autoclaving at 121° C. for 30 minutes. The solids content and the amount of sludges harvested make it possible, from the culture volume centrifuged, to determine the cell concentration (in g/l) of the corresponding culture.

(14) Summarizing table for the tests:

(15) TABLE-US-00005 Solvent for reconstituting Amount the Culture of Solids concentrated volume sludges content [cell] Absorbance culture medium (l) (g) (%) (g/l) (600 nm) Osmotically 10.98 220 5.41 1.08 3.17 treated water Mineral water 7.938 415 2.37 1.24 4.39 Vittel

(16) Conclusion:

(17) In comparison with the control culture on a medium reconstituted with osmotically treated water, the use of water from Vittel improves the growth of the bacterium; the cell yield is increased by 13%.

EXAMPLE 5

Composition Comprising a Vitreoscilla filiformis Extract

(18) Phases A, B, C and D are prepared and successively mixed:

(19) TABLE-US-00006 Amount Phase (g) A PEG/PPG-18/18 dimethicone 10 A Polysorbate 20 0.5 A Cyclopentasiloxane 17 A Cetyl dimethicone 6 A Tocopherol 0.1 A Preservatives 0.7 B Water 25 B Magnesium sulphate 1 B Glycerol 7 B Propylene glycol 3 C Water 18.4 D Water 10 D Vitreoscilla filiformis extract* 1 *cultured according to Example 1 but on a medium comprising 1% of water from La Roche-Posay, the remainder being osmotically treated water.

(20) The composition thus obtained can be applied morning and/or evening to the face, neck, hands and/or the whole of the body.