U.S. patent application number 10/408357 was filed with the patent office on 2003-12-04 for administration of extracts of nonfruiting nonphotosynthetic filamentous bacteria for increasing the endogenous synthesis of superoxide dismutase.
This patent application is currently assigned to SOCIETE L'OREAL S.A.. Invention is credited to Mahe, Yann, Meybeck, Alain.
Application Number | 20030224077 10/408357 |
Document ID | / |
Family ID | 29587386 |
Filed Date | 2003-12-04 |
United States Patent
Application |
20030224077 |
Kind Code |
A1 |
Mahe, Yann ; et al. |
December 4, 2003 |
Administration of extracts of nonfruiting nonphotosynthetic
filamentous bacteria for increasing the endogenous synthesis of
superoxide dismutase
Abstract
The present invention relates to the cosmetic or pharmaceutical
use of an extract of a nonfruiting nonphotosynthetic filamentous
bacterium in a composition containing a cosmetically or
pharmaceutically acceptable medium, as an agent increasing the
endogenous synthesis of superoxide dismutase, in particular for
preventing and/or limiting the formation of free radicals and/or
removing the free radicals present in cells; in addition, the
invention relates to a composition comprising at least one extract
of Vitreoscilla filiformis and lycopene.
Inventors: |
Mahe, Yann; (Morsang Sur
Orge, FR) ; Meybeck, Alain; (Courbevoie, FR) |
Correspondence
Address: |
BURNS DOANE SWECKER & MATHIS L L P
POST OFFICE BOX 1404
ALEXANDRIA
VA
22313-1404
US
|
Assignee: |
SOCIETE L'OREAL S.A.
Paris
FR
|
Family ID: |
29587386 |
Appl. No.: |
10/408357 |
Filed: |
April 8, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60371715 |
Apr 12, 2002 |
|
|
|
Current U.S.
Class: |
424/780 |
Current CPC
Class: |
A61K 31/01 20130101;
A61K 38/44 20130101; A61Q 19/004 20130101; A61K 8/9728 20170801;
A61Q 19/08 20130101; A61K 45/06 20130101; A61K 9/0014 20130101;
C12Y 111/01006 20130101; A61Q 19/007 20130101; A61K 31/739
20130101; A61K 35/74 20130101; C12Y 115/01001 20130101; A61K
2800/522 20130101; C12Y 111/01 20130101; A61K 38/446 20130101; A61Q
19/00 20130101; A61K 2800/75 20130101; A61P 17/00 20180101; A61Q
7/00 20130101; A61K 8/9706 20170801; A61Q 17/04 20130101; A61K 8/99
20130101; A61K 8/31 20130101; A61K 8/66 20130101; A61K 31/01
20130101; A61K 2300/00 20130101; A61K 31/739 20130101; A61K 2300/00
20130101; A61K 35/74 20130101; A61K 2300/00 20130101; A61K 38/446
20130101; A61K 2300/00 20130101; A61K 38/44 20130101; A61K 2300/00
20130101 |
Class at
Publication: |
424/780 |
International
Class: |
A61K 035/70 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 8, 2002 |
FR |
02/04339 |
Claims
What is claimed is:
1. A regime or regimen for increasing the endogenous synthesis of
superoxide dismutase in an individual subject in need of such
treatment, comprising administering to such individual subject a
thus effective amount of at least one extract of at least one
nonfruiting nonphotosynthetic filamentous bacterium.
2. A regime or regimen for preventing and/or limiting the formation
of free radicals and/or eliminating free radicals present in cells
of an individual subject in need of such treatment, comprising
administering to such individual subject a thus effective amount of
at least one extract of at least one nonfruiting nonphotosynthetic
filamentous bacterium.
3. A regime or regimen for preventing and/or combating the harmful
effects of UV radiation and/or pollution on the skin, comprising
administering to an individual subject in need of such treatment, a
thus effective amount of at least one extract of at least one
nonfruiting nonphotosynthetic filamentous bacterium.
4. A regime or regimen for preventing and/or treating the loss of
firmness and/or of elasticity of the skin, comprising administering
to an individual subject in need of such treatment, a thus
effective amount of at least one extract of at least one
nonfruiting nonphotosynthetic filamentous bacterium.
5. A regime or regimen for preventing and/or treating dull skin
complexion, comprising administering to an individual subject in
need of such treatment, a thus effective amount of at least one
extract of at least one nonfruiting nonphotosynthetic filamentous
bacterium.
6. A regime or regimen for preventing and/or treating skin
dehydration, comprising administering to an individual subject in
need of such treatment, a thus effective amount of at least one
extract of at least one nonfruiting nonphotosynthetic filamentous
bacterium.
7. A regime or regimen for preventing and/or treating the signs of
epidermal skin aging, comprising administering to an individual
subject in need of such treatment, a thus effective amount of at
least one extract of at least one nonfruiting nonphotosynthetic
filamentous bacterium.
8. A regime or regimen as defined by claim 7, said signs of
epidermal skin aging comprising pigmented spots and/or
hyperkeratosis spots and/or epidermal atrophy and/or skin roughness
and/or skin drying.
9. A regime or regimen for preventing and/or limiting and/or
eliminating the peroxidation of skin lipids, comprising
administering to an individual subject in need of such treatment, a
thus effective amount of at least one extract of at least one
nonfruiting nonphotosynthetic filamentous bacterium.
10. A regime or regimen for preventing and/or limiting and/or
eliminating objectionable body odor, comprising administering to an
individual subject in need of such treatment, a thus effective
amount of at least one extract of at least one nonfruiting
nonphotosynthetic filamentous bacterium.
11. A regime or regimen for photoprotecting the skin against the
damaging effects of solar UV radiation, comprising topically
applying thereon a thus effective amount of at least one extract of
at least one nonfruiting nonphotosynthetic filamentous
bacterium.
12. A regime or regimen for attenuating and/or repairing skin
redness and/or skin irritation and/or sensations of overheating of
the skin caused by solar radiation, comprising administering to an
individual subject in need of such treatment, a thus effective
amount of at least one extract of at least one nonfruiting
nonphotosynthetic filamentous bacterium.
13. A regime or regimen for revitalizing the scalp and improving
the appearance of the hair, comprising topically applying thereon a
thus effective amount of at least one extract of at least one
nonfruiting nonphotosynthetic filamentous bacterium.
14. A regime or regimen for revitalizing the nails, comprising
topically applying thereon a thus effective amount of at least one
extract of at least one nonfruiting nonphotosynthetic filamentous
bacterium.
15. The regime or regimen as defined by claim 1, said at least one
extract of at least one nonfruiting nonphotosynthetic filamentous
bacterium comprising a lipopolysaccharide isolate.
16. The regime or regimen as defined by claim 1, said at least one
nonfruiting nonphotosynthetic filamentous bacterium comprising
Vitreoscilla filiformis.
17. The regime or regimen as defined by claim 1, comprising
increasing the endogenous synthesis of superoxide dismutase SOD
type 2 (MnSOD).
18. The regime or regimen as defined by claim 1, comprising
coadministering an effective amount of at least one antioxidant to
such individual subject.
19. The regime or regimen as defined by claim 18, said at least one
antioxidant comprising lycopene.
20. The regime or regimen as defined by claim 18, said at least one
antioxidant comprising superoxide dismutase.
21. The regime or regimen as defined by claim 1, comprising
topically applying said effective amount of at least one extract of
at least one nonfruiting nonphotosynthetic filamentous bacterium
onto the skin of such individual subject.
22. A regime or regimen for treating skin erythema or skin oedema,
comprising topically applying onto the afflicted skin of an
individual subject in need of such treatment, a thus effective
amount of at least one extract of at least one nonfruiting
nonphotosynthetic filamentous bacterium.
23. A regime or regimen for treating psoriasis, comprising
topically applying onto the afflicted skin of an individual subject
in need of such treatment, a thus effective amount of at least one
extract of at least one nonfruiting nonphotosynthetic filamentous
bacterium.
24. A regime or regimen for inhibiting the development of cancerous
skin tumors, comprising topically applying onto the skin of an
individual subject in need of such treatment, a thus effective
amount of at least one extract of at least one nonfruiting
nonphotosynthetic filamentous bacterium.
25. A regime or regimen for preventing and/or limiting and/or
eliminating the oxidation phenomena caused by colonization of the
skin by microorganisms, comprising topically applying onto the
afflicted skin of an individual subject in need of such treatment,
a thus effective amount of at least one extract of at least one
nonfruiting nonphotosynthetic filamentous bacterium.
26. A topically applicable cosmetic/dermatological composition
suited for increasing the endogenous synthesis of superoxide
dismutase, comprising a thus effective amount of at least one
extract of at least one nonfruiting nonphotosynthetic filamentous
bacterium, formulated into a topically applicable, physiologically
acceptable medium therefor.
27. The cosmetic/dermatological composition as defined by claim 26,
further comprising an effective amount of at least one
antioxidant.
28. The cosmetic/dermatological composition as defined by claim 27,
said at least one antioxidant comprising lycopene.
29. The cosmetic/dermatological composition as defined by claim 26,
further comprising an effective amount of at least one
catalase.
30. The cosmetic/dermatological composition as defined by claim 26,
further comprising an effective amount of at least one
peroxidase.
31. The cosmetic/dermatological composition as defined by claim 26,
further comprising effective amounts of at least one catalase and
at least one peroxidase.
32. The cosmetic/dermatological composition as defined by claim 26,
comprising an effective amount of at least one extract of
Vitreoscilla filiformis.
33. The cosmetic/dermatological composition as defined by claim 26,
further comprising an effective amount of at least one UV
sunscreen.
34. The cosmetic/dermatological composition as defined by claim 26,
further comprising an effective amount of at least one desquamating
and/or moisturizing agent; depigmenting or propigmenting agent;
antiglycation agent; agent stimulating the synthesis of dermal or
epidermal macromolecules and/or preventing degradation thereof;
agent stimulating the proliferation of fibroblasts and/or
keratinocytes or stimulating the differentiation of keratinocytes;
muscle relaxant; antipollution and/or anti-free radical agent;
slimming agent; agent acting on the microcirculation; agent acting
on the energy metabolism of the cells; tightening agent; or mixture
thereof.
35. The cosmetic/dermatological composition as defined by claim 26,
further comprising an effective amount of at least one
.alpha.-hydroxy acid; salicyclic acid or derivative thereof; HEPES;
procysteine; O-octanoyl-6-D-maltose; disodium salt of
methylglycinediacetic acid; ceramide; steroid; kojic acid;
N-ethyloxycarbonyl- 4-para-aminophenol; ascorbic acid or derivative
thereof; bilberry extract; retinoid; polypeptide or acylated
derivative thereof; phytohormone; extract of the yeast
Saccharomyces cerevisiae; extract of algae; extract of soyabean,
lupin, maize and/or pea; alverine or salt thereof; resveratrol;
carotenoid; tocopherol or ester thereof; coenzyme Q10 or
ubiquinone; xanthins; extract of butcher's broom or of horse
chestnut; or mixture thereof.
36. The cosmetic/dermatological composition as defined by claim 26,
formulated as a solution, emulsion, gel, paste, solid, cream,
ointment, milk, serum, lotion, mousse, or aerosol.
37. The cosmetic/dermatological composition as defined by claim 28,
comprising from 0.001% to 10% by dry weight of said at least one
extract of at least one nonfruiting nonphotosynthetic filamentous
bacterium and from 10.sup.-9% to 0.1% by weight of lycopene.
Description
CROSS-REFERENCE TO PRIORITY/PROVISIONAL APPLICATIONS
[0001] This application claims priority under 35 U.S.C. .sctn. 119
of FR-02/04339, filed Apr. 8, 2002, and of provisional application
Serial No. 60/371,715, filed Apr. 11, 2002, both hereby expressly
incorporated by reference. This application is also a continuation
of said '715 provisional.
BACKGROUND OF THE INVENTION
[0002] 1. Technical Field of the Invention
[0003] The present invention relates to the cosmetic or
pharmaceutical use of an extract of a nonfruiting (nonfructifying)
nonphotosynthetic filamentous bacterium in a composition containing
a cosmetically or pharmaceutically acceptable medium, as an agent
increasing the endogenous synthesis of superoxide dismutase (SOD),
in particular for preventing and/or limiting the formation of free
radicals and/or removing the free radicals present in cells.
[0004] 2. Description of the Prior Art
[0005] Over time, various signs appear on the skin which are very
characteristic of aging, resulting in particular in a modification
of the structure and functions of the skin.
[0006] This aging, which is of a physiological nature, results from
the action of two main classes of components, an endogenous
component resulting in particular from the natural production of
superoxide ions, in particular produced during cell respiration.
The other component is exogenous. Indeed, aging may be accelerated
by environmental factors such as repeated exposure of the skin to
sunlight, and in particular to ultraviolet A radiation, to
pollution, in particular to diesel particles and to cigarette
smoke.
[0007] It is known that the toxicity of atmospheric pollutants, in
particular of gaseous pollutants such as sulphur dioxide, ozone and
nitrogen oxides on the constituents of the skin (fibers, cells,
enzymes) and on the sebum secreted by the skin is linked in
particular to their free radical initiating activity, a source of
oxidation phenomena which cause cellular damage in living
beings.
[0008] Living cells, which are in direct and permanent contact with
the external medium (in particular the skin, the scalp and certain
mucous membranes) are particularly sensitive to these effects of
gaseous pollutants, which result in particular in accelerated aging
of the skin, with a complexion which lacks radiance and an early
formation of wrinkles or fine lines, and also in a decrease in the
vitality and a dull appearance of the hair.
[0009] It is also known that the irradiation phenomena caused by
exposure to ultraviolet rays also lead to the phenomena of
accelerated cellular aging.
[0010] Whether they are of endogenous or exogenous origin, free
radicals cause substantial oxidative damage, in particular in the
cell membranes (peroxidation of lipids causing a deterioration of
the permeability of membranes), the cell nuclei (destruction of
DNA) and the tissues, in particular the connective tissue
(degradation of the elastin and collagen fibers, depolymerization
of the polyuronic fibers). This damage leads in particular to
drying and a loss of firmness and of elasticity of the skin
(Grinwald et al., 1980, Agren et al., 1997).
[0011] Specialists currently consider that one of the causes of
cellular aging is the reduction in the defense capabilities against
free radicals and against oxidation phenomena (in particular the
formation of superoxide ions) which they initiate.
[0012] The superoxide ion O.sup.o-.sub.2 (active oxygen) is a
radical ion whose instability and reactivity make it a toxic
compound, because it generates, in particular in the presence of
metal ions, highly harmful hydroxyl free radicals (OH.sup.o).
Superoxide dismutases (SOD) are enzymes which exert a protective
effect in particular by trapping the superoxide ions and thus
constitute a biological system of defence against the harmful
effects of free radicals.
[0013] Superoxide dismutases are capable of inducing the
dismutation of the superoxide ions, according to the reaction:
2O.sup.o-.sub.2+2H.sup.+.fwdarw.H.sub.2O.sub.2+O.sub.2
[0014] Numerous superoxide dismutases are known. For example,
superoxide dismutases extracted from bovine erythrocytes
(Markovitz, J. Biol. Chem. 234, p. 40, 1959) and superoxide
dismutases extracted from Escherichia coli (Keele and Fridovich, J.
Biol. Chem., 245, p. 6176, 1970) have already been described. In
the document FR-A-2,225,443 are described superoxide dismutases
extracted from marine bacterial strains, and their method of
preparation.
[0015] Superoxide dismutases make it possible in particular to
protect the skin and the hair, in particular by maintaining the
integrity of the natural keratin structure, as describes for
example the document FR-A-2,287,899. In addition, superoxide
dismutases improve cutaneous cell respiration and maintain or
improve the qualities of the skin, such as softness to the touch,
suppleness and elasticity.
[0016] Superoxide dismutases also protect the skin against the
inflammation phenomena caused by ultraviolet radiation and against
accelerated skin aging, in particular under the influence of such
radiation.
[0017] Because of these advantageous properties, it is known to add
superoxide dismutases to cosmetic compositions, in particular
compositions intended for topical application (see, for example,
EP-0-673,643 and EP-0-636,360).
SUMMARY OF THE INVENTION
[0018] The Applicant has discovered a novel means of combating the
harmful effects caused by free radicals by inducing the endogenous
synthesis of superoxide dismutase. Indeed, it has discovered, most
surprisingly, that an extract of a nonfruiting and
nonphotosynthetic filamentous bacterium caused an increase in the
endogenous synthesis of superoxide dismutase.
[0019] The topical use of substances with superoxide dismutase
activity, while it can be advantageous under certain conditions of
use, exhibits nevertheless the disadvantage of requiring particular
stability and cutaneous bioavailability of these SOD forms.
Moreover, it is known that a particular distribution of SODs exists
in cells, in particular they are physiologically located inside the
cells and more particularly in direct proximity with cellular
entities which strongly generate superoxide ions, such as the
mitochondria. Thus, when means are used for neutralizing the
superoxide ions produced outside the cell, it is because the
oxidative cellular damage is already very extensive and the
superoxide anion has left the control of its intracellular
environment. The use of exogenous SOD is therefore designed to
prevent the propagation of oxidative damage to other cellular
entities which can fulfil both a curative and repair function.
[0020] The Applicant presents here a means of inducing the cells to
produce their own antioxidant activity and to thus strengthen their
natural defenses. The advantage of the use according to the present
invention compared with the use of an exogenous substance
exhibiting an SOD activity is that it makes it possible, after
contact between the cells and the membrane extract of nonfruiting
nonphotosynthetic filamentous bacterium, to induce the production
of the SOD enzyme according to a localization (in particular
mitochondrial) which respects the physiological conditions and in
particular induces the cell to protect itself from inside by virtue
of its own oxidative metabolism. Furthermore, SOD, when it captures
free radicals, itself undergoes oxidation and gradually loses its
resistance to oxidative damage, this is even more manifest when the
enzyme has left its intracellular context. The induction of SOD in
response to the application of an extract of a nonfruiting
nonphotosynthetic filamentous bacterium thus allows the cell to
self-reconstruct and to accelerate the physiological renewal of its
intracellular stock of SOD.
[0021] Increasing the endogenous synthesis of superoxide dismutases
has numerous advantages compared with the exogenous supply of the
enzyme. Indeed, during topical application of compositions
containing SODs, a large proportion of SODs are denatured in
particular by proteases present at the surface of the skin and/or
not crosslinking the cell membranes because of their large
molecular weight (17 kD for Cu/ZnSOD and 23 kD for MnSOD).
Accordingly, to obtain the desired effect, it is generally
necessary to increase the quantity of SOD in the composition.
[0022] Another disadvantage of the topical application of an
exogenous protein, such as the SOD enzyme, is the risk of allergy
which it represents when the protein concentration is too high.
[0023] The subject of the present invention is therefore the
cosmetic use of an extract of a nonfruiting nonphotosynthetic
filamentous bacterium in a composition containing a cosmetically
acceptable medium, as an agent increasing the endogenous synthesis
of superoxide dismutase.
DETAILED DESCRIPTION OF BEST MODE AND SPECIFIC/PREFERRED
EMBODIMENTS OF THE INVENTION
[0024] The bacterial extracts according to the invention are
prepared from nonfruiting and nonphotosynthetic filamentous
bacterium as defined according to the classification of Bergey's
Manual of Systematic Bacteriology (Vol. 3, section 23, 9th Edition,
1989), among which there may be mentioned bacteria belonging to the
order Beggiatoales, and more particularly bacteria belonging to the
genera Beggiatoa, Vitreoscilla, Flexithrix or Leucothrix.
[0025] Various extracts may be used, in particular it is preferable
to use, as an extract of a nonfruiting nonphotosynthetic
filamentous bacterium, a lipopolysaccharide isolate which may, for
example, be obtained according to one of the methods described in
Example 1.
[0026] The preferred nonfruiting nonphotosynthetic filamentous
bacterium according to the present invention is Vitreoscilla
filiformis, in particular the strain ATCC 15551.
[0027] Human cells synthesize two types of superoxide dismutases,
superoxide dismutase type 1, also called Cu/ZnSOD or SOD of the
cytosol, is an enzyme which is found in the cytosol of cells.
Superoxide dismutase type 2, MnSOD, is found in the mitochondria of
the cells. The Applicant has observed, most surprisingly, that the
extract of a nonfruiting nonphotosynthetic filamentous bacterium
was particularly effective for increasing the synthesis of MnSOD by
a transcriptional route involving an increase in the production of
messenger RNA, a precursor for its protein synthesis.
[0028] Accordingly, a variant of the invention relates to the
cosmetic use of an extract of a nonfruiting nonphotosynthetic
filamentous bacterium in a composition containing a cosmetically
acceptable medium, as an agent increasing the endogenous synthesis
of superoxide dismutase type 2 (MnSOD).
[0029] In the compositions according to the invention, it is
possible to use 0.001% to 10%, and in particular from 0.01% to 1%,
by weight of dry extract of nonfruiting nonphotosynthetic
filamentous bacterium relative to the weight of the
composition.
[0030] In particular forms of applications of the balneotherapy
type, it is also possible to envisage applications of native or
reconstituted bacterial lysates in higher proportions which may be
up to 100%.
[0031] These compositions may contain the Vitreoscilla filiformis
extract in the form of a dispersion in an appropriate vehicle such
as, for example, water, organic solvents, fatty substances
including oils, and mixtures thereof, in particular emulsions.
[0032] According to another subject of the present invention and
for all the applications described below, during the cosmetic use
of the extract of a nonfruiting nonphotosynthetic filamentous
bacterium in a composition containing a cosmetically acceptable
medium, the extract of a nonfruiting nonphotosynthetic filamentous
bacterium may be combined with an antioxidant.
[0033] Thus, it is possible, for example, to use an antioxidant
chosen from:
[0034] vitamin E (tocopherol) and its derivatives, inter alia
acetate, linoleate or nicotinate, preferably at concentrations of
the order of 0.1% to 5%,
[0035] .gamma.-orizanol (0.1% to 5%),
[0036] lysine or arginine pidolates (0.5% to 10%),
[0037] plant extracts such as Melissa extract (0.01% to 2%),
silimarin extract (0.01% to 2%), gingko biloba extract (0.05% to
2%), sage extract (0.05% to 2%), kola nut extract (0.05% to 2%),
rutin extract (0.1% to 2%) or thyme extract (0.1% to 2%), the %
being given as dry matter,
[0038] lycopene in purified form or alternatively in an extract
(for example tomato paste having a lycopene titre resulting in a
final lycopene concentration of between 10.sup.-12% and 10%, and
more preferably from 10.sup.-7% to 0.1%),
[0039] pine, hawthorn or grape proanthocyanolidic oligomers (0.1%
to 2%),
[0040] di-tert-butyl-hydroxybenzylidenecamphor (0.1% to 2%),
[0041] green tea (0.1% to 2%),
[0042] caffeine (0.1% to 5%),
[0043] glycerol (2% to 30%),
[0044] mannitol (2% to 30%),
[0045] carnosine (0.1% to 2%),
[0046] superoxide dismutase (100 to 10 000 IU/100 g),
[0047] guanosine (0.01% to 1%),
[0048] microalgae containing ethoxyquin such as Hematococcus
(0.005% to 1%),
[0049] pentasodium aminotrimethylene phosphonate (0.001% to
0.5%),
[0050] lactoperoxydase (0.01% to 0.1%), and
[0051] lactoferrin (0.01% to 0.1%).
[0052] It is also possible to use a mixture of several
antioxidants.
[0053] There may also be mentioned anti-free radical agents, in
particular biflavonoids, coenzyme Q10 or ubiquinone; certain
enzymes such as catalase, glutathione peroxidase and quinine
reductases; glutathione; benzylidenecamphor; benzylcyclanones;
substituted naphthalenones; pidolates; phytantriol; lignans; and
melatonin.
[0054] Preferably, the antioxidant is lycopene.
[0055] In another embodiment according to the invention, the
antioxidant is a superoxide dismutase. It will be possible, for
example, to use the SOD enzyme extracted from bovine erythrocytes
(Markovitz, J. Biol. Chem. 234, p. 40, 1959), from Escherichia coli
(Keele and Fridovich, J. Biol. Chem., 245, p. 6176, 1970) or
alternatively from marine bacterial strains (FR-A-2,225,443).
[0056] The present invention also relates to the cosmetic use of an
extract of a nonfruiting nonphotosynthetic filamentous bacterium in
a composition containing a cosmetically acceptable medium, for
preventing and/or limiting the formation of free radicals and/or
removing the free radicals present in cells.
[0057] Another subject to the invention relates to cosmetic use of
an extract of a nonfruiting nonphotosynthetic filamentous bacterium
in a composition containing a cosmetically acceptable medium, for
preventing and/or combating the harmful effects of UV radiation
and/or of pollution on the skin.
[0058] Clinically, the harmful effects of UV radiation and/or of
pollution on the skin generally result in accelerated aging, that
is to say in the appearance of wrinkles and fine lines, in a
loosening of the cutaneous and subcutaneous tissues, in a loss of
cutaneous elasticity and in skin texture atony. The loss of
firmness and of tonicity of the skin, such as wrinkles and fine
lines, is explained, at least in part, by dermal atrophy and
flattening of the dermal/epidermal junction; the skin is less firm
and more flaccid, and the thickness of the epidermis decreases.
[0059] In addition, the complexion of the skin is generally
modified, it appears more pale and more yellow. This phenomenon
appears to be essentially due to disorganization of the
microcirculation (less haemoglobin in the papillary dermis).
Another clinical sign of aging is the dry and rough appearance of
the skin which is essentially due to greater desquamation; these
squamas, by diffracting the light rays, also participate in the
slightly grey appearance of the complexion. Furthermore, numerous
colored and/or dark spots appear at the surface of the skin, and
more especially on the hands, conferring heterogeneity on the skin.
In general, these spots are due to the high production of melanin
in the epidermis and/or the dermis of the skin. Moreover, diffuse
irritations and sometimes telagiectasias may exist on certain areas
of the skin. Some of these signs are more particularly linked to
intrinsic or physiological aging, that is to say aging linked to
age, whereas others are more specific to extrinsic aging, that is
to say aging caused in general by the environment; it is more
particularly photoaging due to exposure to the sun, light or any
other radiation.
[0060] Accordingly, the subject of the invention is more
particularly suited to the cosmetic use of an extract of a
nonfruiting nonphotosynthetic filamentous bacterium in a
composition containing a cosmetically acceptable medium, for
preventing and/or treating the loss of firmness and/or elasticity
of the skin. Such a use allows in particular the skin to rediscover
a uniformly smooth appearance.
[0061] Another subject of the invention is the cosmetic use of an
extract of a nonfruiting nonphotosynthetic filamentous bacterium in
a composition containing a cosmetically acceptable medium, for
preventing and/or treating dull complexion.
[0062] The invention is also suited to the cosmetic use of an
extract of a nonfruiting nonphotosynthetic filamentous bacterium in
a composition containing a cosmetically acceptable medium, for
preventing and/or treating skin dehydration.
[0063] More generally, the subject of the invention is also suited
to the cosmetic use of an extract of a nonfruiting
nonphotosynthetic filamentous bacterium in a composition containing
a cosmetically acceptable medium, for preventing and/or treating
the signs of skin aging.
[0064] The expression signs of skin aging is understood more
particularly to mean pigmented spots and/or hyperkeratosis spots
and/or epidermal atrophy and/or skin roughness and/or skin
dryness.
[0065] As explained above, an undesirable effect of the presence of
free radicals in the skin is that they cause a phenomenon of
peroxidation of lipids. With age (more particularly from forty
years), the accumulation of these peroxidized lipids is responsible
for bad body odors such as a rancid odor (Haze S. et. al. J.
Invest. Dermatol. 2001, 116(4): 520-4).
[0066] The subject of the invention is suited to the cosmetic use
of an extract of a nonfruiting nonphotosynthetic filamentous
bacterium in a composition containing a cosmetically acceptable
medium, for preventing and/or limiting and/or eliminating the
peroxidation of skin lipids.
[0067] Accordingly, the subject of the invention is also useful for
preventing and/or limiting and/or eliminating bad body odours.
[0068] In the particular case of exposure of the skin to the sun,
it appears that moderate exposure to UVA and UVB induces, in the
first instance, a reduction in skin SODs (Leccia et al., Exp.
Dermatol. 2001, 10(4): 272-9). Five days after this exposure, a
rebound effect is observed with an increase in the activity of the
SODs. Thus, a transitional period is necessary for putting in place
a skin antioxidant protective system.
[0069] By virtue of its capacity to increase the endogenous
synthesis of SOD, the extract of a nonfruiting nonphotosynthetic
filamentous bacterium makes it possible to accelerate the putting
in place of the skin antioxidant protective system and to prepare
the skin for solar exposure.
[0070] Accordingly, the cosmetic use according to the invention of
an extract of a nonfruiting nonphotosynthetic filamentous bacterium
in a composition containing a cosmetically acceptable medium is
particularly well suited for preparing the skin for solar
exposure.
[0071] In particular, the preparation of the skin for solar
exposure may be carried out by daily application, to the skin, of
the said cosmetic composition for one week before the solar
exposure and, preferably, for two weeks, up to at least one night
(between 6 and 18 hours) before the solar exposure.
[0072] The use according to the present invention is also useful
during and after solar exposure for maintaining a high level of SOD
synthesis and to attenuate and/or repair the damage linked to solar
exposure such as redness, skin irritations and sensations of
overheating of the skin.
[0073] The subject of the invention may also consist in the
cosmetic use of an extract of a nonfruiting nonphotosynthetic
filamentous bacterium in a composition containing a cosmetically
acceptable medium for attenuating and/or repairing the redness
and/or the skin irritations and/or the sensations of overheating of
the skin caused by solar exposure.
[0074] Preferably, the cosmetic composition used according to the
present invention is suited to topical application.
[0075] The use of an extract of a nonfruiting nonphotosynthetic
filamentous bacterium in a cosmetic composition according to the
invention may also be provided in the form of a hair tonic for
revitalizing the scalp and for improving the appearance of the
hair.
[0076] In another variant of the invention, the use of an extract
of a nonfruiting nonphotosynthetic filamentous bacterium in a
cosmetic composition is applied to the nails in order to revitalize
the nails. In such an application, the cosmetic composition may for
example be provided in the form of a varnish or a gel or a cream
for massaging the nails.
[0077] Another subject of the present invention relates to the use
of at least one extract of a nonfruiting nonphotosynthetic
filamentous bacterium for the preparation of a pharmaceutical or
dermatological composition intended for increasing the endogenous
synthesis of superoxide dismutase.
[0078] Thus, the use of at least one extract of a nonfruiting
nonphotosynthetic filamentous bacterium may also be useful for the
preparation of a dermatological composition intended for preventing
and/or limiting the formation of free radicals and/or for
eliminating the free radicals present in cells.
[0079] In particular, the present invention relates to the use of
at least one extract of a nonfruiting nonphotosynthetic filamentous
bacterium for the preparation of a dermatological composition
intended for repairing the damage caused by solar exposure, in
particular when the damage caused by solar exposure is a skin
erythema or a skin oedema.
[0080] The present invention also relates to the use of at least
one extract of a nonfruiting nonphotosynthetic filamentous
bacterium for the preparation of a dermatological composition
intended for preventing and/or limiting and/or eliminating the
oxidation phenomena caused by the colonization of the skin by
microorganisms. Preferably, the invention relates to the use of at
least one extract of a nonfruiting nonphotosynthetic filamentous
bacterium for the preparation of a dermatological composition
intended for treating acne.
[0081] It has been shown that the tissues of a psoriatic skin
highly expressed mRNAs encoding MnSOD and that this phenomenon
represented a response designed to protect the cells (Lontz et al.,
Free Radic. Biol. Med. 1995; 18(2): 349-55). By increasing the
endogenous synthesis of MnSOD, the use according to the invention
contributes to the treatment of psoriatic skins. Accordingly, the
subject of the invention is suited to the use of at least one
extract of a nonfruiting nonphotosynthetic filamentous bacterium
for the preparation of a dermatological composition intended for
treating psoriasis.
[0082] Moreover, a study on a model of carcinogenesis of the skin
comparing the progression of tumours in transgenic mice expressing
MnSOD in the skin and in wild-type mice has shown that the
expression of MnSOD had as a consequence the inhibition of the
development of tumours (Zhao et al., Cancer Res. 2001, 61(16):
6082-8). Accordingly, the subject of the invention is suited to the
use of at least one extract of a nonfruiting nonphotosynthetic
filamentous bacterium for the preparation of a dermatological
composition intended for inhibiting the development of cancerous
skin tumours.
[0083] The present invention also relates to the use of at least
one extract of a nonfruiting nonphotosynthetic filamentous
bacterium for the preparation of a dermatological composition
intended for preventing and/or treating the signs of epidermal
aging. These signs of skin aging are in particular pigmented spots
and/or hyperkeratosis spots and/or epidermal atrophy and/or skin
roughness and/or skin dryness.
[0084] In another variant, the present invention relates to a
composition containing, in a physiologically acceptable medium at
least one extract of a nonfruiting nonphotosynthetic filamentous
bacterium, preferably an extract of Vitreoscilla filiformis, and
lycopene.
[0085] Lycopene is a natural pigment which is found in ripe fruits,
particularly in tomatoes. It belongs to the family of carotenoids
and its structure is close to that of .beta.-carotene.
[0086] The role of lycopene in the maturation of fruits is known in
the prior art.
[0087] Lycopene is used in compositions with tanning activity for
its role on the synthesis of melanin (WO 97/47278), in compositions
intended for the treatment of hair and/or of acne for its activity
on 5.alpha.-reductases (JP-2940964) or alternatively as anti-free
radical agent (JP-A-8-283136).
[0088] Lycopene is also described as an inhibitor of the expression
of proteases of the extracellular matrix, particularly
metalloproteinases, such as for example collagenases
(EP-A-1,090,628).
[0089] Lycopene may be in the cis or trans chemical form.
[0090] To give an order of magnitude, pure lycopene may be used in
a quantity representing from 10.sup.-9% to 0.1% of the total weight
of the composition, and preferably in a quantity representing from
10.sup.-7% to 10.sup.-3% of the total weight of the
composition.
[0091] To give another order of magnitude, some tomato pastes sold
on the market, such as Lyc-O-Derm.RTM. (Lycored) have a pure
lycopene titre of 10%; it is therefore possible to use a quantity
of pasty tomato extract in proportions ranging from 10.sup.-6% to
10.sup.-2% of tomato extract.
[0092] In the particular applications of balneotherapy, it is
possible to envisage contact with the skin in proportions where the
tomato extract may be applied pure and may reach lycopene titres
applied to the skin or the mucous membranes which may range from 1%
to 15%.
[0093] The subject of the present invention is also a composition
containing, in a physiologically acceptable medium, at least one
extract of Vitreoscilla filiformis and a compound having a catalase
activity.
[0094] As compounds which have a catalase activity, it is possible
to use in particular catalases of natural (plant or animal) origin
or catalases which have been modified chemically or by grafting, by
adsorption onto supports, or by encapsulation (see in particular
Patent Applications FR-2-716,884 and GB-793,739).
[0095] It is also possible to use commercial catalases such as
Catalase NL.RTM. which are sold by the company Amano Enzyme Europe
Ltd.
[0096] The subject of the present invention may also be a
composition containing, in a physiologically acceptable medium, at
least one extract of Vitreoscilla filiformis and a compound having
a peroxidase activity.
[0097] As compounds having a peroxidase activity, it is possible to
use in particular peroxidases of natural (plant or animal) origin,
or alternatively peroxidases which have been modified chemically or
by grafting, by adsorption onto supports, or by encapsulation (see
for example WO 01/46431 and WO 87/07838 and EP-A-0,397,227).
[0098] It is possible to use in particular lactoperoxidases,
microperoxidases from fungi, myeloperoxidase, and the like. It is
known that lactoperoxidase (abbreviated: LPO) is an enzyme which is
found in particular in numerous mammalian tissues and secretions,
which uses one of the numerous cellular electron donors to reduce
organic peroxides of the ROOH type (R being an organic group).
Lactoperoxidase is a commercial product sold in particular by the
companies Sigma and Sederma.
[0099] It may also be possible to use recombinant peroxidases, for
example recombinant LPO (WO 91/06639).
[0100] Finally, the composition according to the invention may
contain, in a physiologically acceptable medium, at least one
extract of Vitreoscilla filiformis, a compound having a catalase
activity and a compound having a peroxidase activity.
[0101] The invention also relates to a method for the cosmetic
treatment of the loss of firmness and/or of elasticity of the skin,
comprising the application, to the skin, of a composition
containing, in a cosmetically acceptable medium, at least one
extract of Vitreoscilla filiformis.
[0102] The invention relates to a method of cosmetic treatment for
preparing the skin for solar exposure, comprising the application
to the skin of a composition containing, in a cosmetically
acceptable medium, at least one extract of Vitreoscilla
filiformis.
[0103] The compositions used according to the invention may be
provided in all the forms which may be envisaged in the cosmetic
and pharmaceutical, in particular dermatological, field.
[0104] The composition according to the invention is preferably
suited to topical application to the skin. It may be provided in
all the galenic forms normally used for this type of application,
in particular in the form of an aqueous or oily solution, an
oil-in-water or water-in-oil or multiple emulsion, a silicone
emulsion, a macroemulsion or nanoemulsion, an aqueous or oily gel
or a liquid, pasty or solid anhydrous product.
[0105] This composition may be more or less fluid and may have the
appearance of a white or coloured cream, an ointment, a milk, a
lotion, a serum, a paste, a mousse or a gel. It may optionally be
applied to the skin in the form of an aerosol. It may also be
provided in solid form, for example in the form of a stick. It may
be used as a care product and/or as a make-up product for the
skin.
[0106] It may even be, in applications in the same category as
balneotherapy, a crude extract.
[0107] To reinforce the anti-aging effects of the composition
according to the invention, it may contain, in addition to the
extract of a nonfruiting nonphotosynthetic filamentous bacterium
described above, at least one compound chosen from: desquamating
and/or moisturizing agents; depigmenting or propigmenting agents;
antiglycation agents; agents stimulating the synthesis of dermal or
epidermal macromolecules and/or preventing their degradation;
agents stimulating the proliferation of fibroblasts and/or
keratinocytes or stimulating the differentiation of keratinocytes;
muscle relaxants; antipollution and/or anti-free radical agents;
slimming agents; agents acting on the microcirculation; agents
acting on the energy metabolism of the cells; tightening agents;
and mixtures thereof.
[0108] Thus, the composition according to the invention may in
particular contain at least one active agent chosen from:
a:-hydroxy acids; salicyclic acid and its derivatives such as
5-(n-octanoyl)salicyclic acid; HEPES; procysteine;
O-octanoyl-6-D-maltose; the disodium salt of methylglycinediacetic
acid; ceramides; steroids such as diosgenin and derivatives of
DHEA; kojic acid; N-ethyloxycarbonyl-4-para-aminophenol; ascorbic
acid and its derivatives; bilberry extracts; retinoids and in
particular retinol and its esters; polypeptides and their acylated
derivatives; phytohormones; extracts of the yeast Saccharomyces
cerevisiae; extracts of algae; extracts of soyabean, lupin, maize
and/or pea; alverine and its salts, in particular alverine citrate;
resveratrol; carotenoids and in particular lycopene; tocopherol and
its esters; coenzyme Q10 or ubiquinone; xanthines and in particular
caffeine and natural extracts containing it; extracts of butcher's
broom and of horse chestnut; and mixtures thereof, without this
list being limiting.
[0109] The composition according to the invention may in addition
contain at least one UVA and/or UVB sun screening agent. The
sunscreens may be chosen from organic screening agents, inorganic
screening agents and mixtures thereof.
[0110] As examples of organic screening agents which are active in
UV-A and/or UV-B, there may be mentioned in particular those
designated below by their CTFA name:
[0111] para-aminobenzoic acid derivatives: PABA, Ethyl PABA, Ethyl
Dihydroxypropyl PABA, Ethylhexyl Dimethyl PABA sold in particular
under the name "ESCALOL 507" by ISP, Glyceryl PABA, PEG-25 PABA
sold under the name "UVINUL P25" by BASF,
[0112] salicyclic derivatives: Homosalate sold under the name
"EUSOLEX HMS" by RONA/EM INDUSTRIES, Ethylhexyl Salicylate sold
under the name "NEO HELIOPAN OS" by HAARMANN and REIMER,
Dipropyleneglycol Salicylate sold under the name "DIPSAL" by SCHER,
TEA Salicylate, sold under the name "NEO HELIOPAN TS" by HAARMANN
and REIMER,
[0113] dibenzoylmethane derivatives: Butyl Methoxydibenzoylmethane
sold in particular under the trademark "PARSOL 1789" by HOFFMANN LA
ROCHE, Isopropyl Dibenzolylmethane,
[0114] cinnamic derivatives: Ethylhexyl Methoxycinnamate sold in
particular under the trademark "PARSOL MCX" by HOFFMANN LA ROCHE,
Isopropyl Methoxy Cinnamate, Isoamyl Methoxy Cinnamate sold under
the trademark "NEO HELIOPAN E 1000" by HAARMANN and REIMER,
Cinoxate, DEA Methoxycinnamate, Diisopropyl Methylcinnamate,
Glyceryl Ethylhexanoate Dimethoxycinnamate,
[0115] .beta.,.beta.-diphenylacrylate derivatives: Octocrylene sold
in particular under the trademark "UVINUL N539" by BASF,
Etocrylene, sold in particular under the trademark "UVINUL N35" by
BASF,
[0116] benzophenone derivatives: Benzophenone-1 sold under the
trademark "UVINUL 400" by BASF, Benzophenone-2 sold under the
trademark "UVINUL D50" by BASF, Benzophenone-3 or Oxybenzone, sold
under the trademark "UVINUL M40" by BASF, Benzophenone-4 sold under
the trademark "UVINUL MS40" by BASF, Benzophenone-5, Benzophenone-6
sold under the trademark "HELISORB 11 " by NORQUAY, Benzophenone-8
sold under the trademark "SPECTRA-SORB UV-24" by AMERICAN CYANAMID,
Benzophenone-9 sold under the trademark "UVINUL DS-49" by BASF,
Benzophenone-12,
[0117] benzylidene camphor derivatives: 3-Benzylidene Camphor,
4-Methylbenzylidene Camphor sold under the name "EUSOLEX 6300" by
MERCK, Benzylidene Camphor Sulphonic Acid, Camphor Benzalkonium
Methosulphate, Terephthalylidene Dicamphor Sulphonic Acid,
Polyacrylamidomethyl Benzylidene Camphor,
[0118] phenylbenzimidazole derivatives: Phenylbenzimidazole
Sulphonic Acid sold in particular under the trademark "EUSOLEX 232"
by MERCK, Benzimidazilate sold under the trademark "NEO HELIOPAN
AP" by HAARMANN and REIMER,
[0119] triazine derivatives: Anisotriazine sold under the trademark
"TINOSORB S" by CIBA GEIGY, Ethylhexyl triazones sold in particular
under the trademark "UVINUL T150" by BASF, Diethylhexyl Butamido
Triazone sold under the trademark "UVASORB HEB" by SIGMA 3V,
[0120] phenylbenzotriazole derivatives: Drometrizole Trisiloxane
sold under the name "SILATRIZOLE" by RHODIA CHIMIE,
[0121] anthranilic derivatives: Menthyl anthranilate sold under the
trademark "NEO HELIOPAN MA" by HAARMANN and REIMER,
[0122] imidazoline derivatives: Ethylhexyl Dimethoxybenzylidene
Dioxoimidazoline Propionate,
[0123] benzalmalonate derivatives: Polyorganosiloxane with
benzalmalonate functional groups sold under the trademark "PARSOL
SLX" by HOFFMANN LA ROCHE, and mixtures thereof.
[0124] The organic UV-screening agents which are more particularly
preferred are chosen from the following compounds:
[0125] Ethylhexyl Salicylate,
[0126] Butyl Methoxydibenzoylmethane,
[0127] Ethylhexyl Methoxycinnamate,
[0128] Octocrylene,
[0129] Phenylbenzimidazole Sulphonic Acid,
[0130] Terephthalylidene Dicamphor Sulphonic,
[0131] Benzophenone-3,
[0132] Benzophenone-4,
[0133] Benzophenone-5,
[0134] 4-Methylbenzylidene camphor,
[0135] Benzimidazilate,
[0136] Anisotriazine,
[0137] Ethylhexyl triazone,
[0138] Diethylhexyl Butamido Triazone,
[0139] Methylene bis-Benzotriazolyl Tetramethylbutylphenol,
[0140] Drometrizole Trisiloxane,
[0141] and mixtures thereof.
[0142] The inorganic screening agents which may be used in the
composition according to the invention are in particular
nanopigments (mean size of the primary particles: generally between
5 nm and 100 nm, preferably between 10 nm and 50 nm) of coated or
uncoated metal oxides such as for example nanopigments of titanium
oxide (amorphous or crystallized in the form of rutile and/or
anatase), iron, zinc, zirconium or cerium oxides. Coating agents
are moreover alumina and/or aluminum stearate. Such nanopigments of
metal oxides, coated or uncoated, are in particular described in
EP-A-0-518,772 and EP-A-0-518,773.
[0143] In a known manner, the composition of the invention may also
contain the customary adjuvants in the cosmetic and dermatological
fields, such as hydrophilic or lipophilic gelling agents,
preservatives, antioxidants, solvents, perfumes, fillers, pigments,
odour absorbers and colouring matters. The quantities of these
various adjuvants are those conventionally used in the fields
considered, and for example from 0.01% to 20% of the total weight
of the composition. These adjuvants, depending on their nature, may
be introduced into the fatty phase or into the aqueous phase. These
adjuvants, and their concentrations, should be such that they do
not damage the advantageous properties of the extract of the
nonfruiting nonphotosynthetic filamentous bacterium.
[0144] As oils which may be used in the composition of the
invention, there may be mentioned for example:
[0145] hydrocarbon oils of animal origin, such as
perhydrosqualene;
[0146] hydrocarbon oils of plant origin, such as the liquid
triglycerides of fatty acids containing from 4 to 10 carbon atoms
and the liquid fraction of shea butter;
[0147] esters and synthetic esters, in particular of fatty acids,
such as the oils of formulae R.sup.1COOR.sup.2 and R.sup.1OR.sup.2
in which R.sup.1 represents the residue of a fatty acid containing
from 8 to 29 carbon atoms, and R.sup.2 represents a hydrocarbon
chain, branched or unbranched, containing from 3 to 30 carbon
atoms, such as for example Purcellin oil, isononyl isononanoate,
isopropyl myristate, 2-ethylhexyl palmitate, 2-octyldodecyl
stearate, 2-octyldodecyl erucate, isostearyl isostearate;
hydroxylated esters such as isostearyl lactate,
octylhydroxystearate, octyldodecyl hydroxystearate, diisostearyl
malate, triisocetyl citrate, heptanoates, octanoates, decanoates of
fatty alcohols ; polyol esters, such as propylene glycol
dioctanoate, neopentylglycol diheptanoate and diethylene glycol
diisononanoate; and pentaerythritol esters such as pentaerythrityl
tetraisostearate;
[0148] linear or branched hydrocarbons, of inorganic or synthetic
origin such as volatile or non-volatile paraffin oils and
derivatives thereof, petroleum jelly, polydecenes, hydrogenated
polyisobutene such as parleam oil;
[0149] fatty alcohols having from 8 to 26 carbon atoms, such as
cetyl alcohol, stearyl alcohol and a mixture thereof (cetylstearyl
alcohol), octyldodecanol, 2-butyloctanol, 2-hexyldecanol,
2-undecylpentadecanol, oleyl alcohol and linoleyl alcohol;
[0150] partially hydrocarbon-based and/or silicone-based
fluorinated oils such as those described in the document
JP-A-2-295912;
[0151] silicone oils such as volatile or non-volatile
polymethylsiloxanes (PDMS) containing a linear or cyclic silicone
which are liquid or pasty at room temperature, in particular
cyclopolydimethylsiloxanes (cyclomethicones) such as
cyclohexasiloxane; polydimethylsiloxanes containing alkyl, alkoxy
or phenyl groups, which are pendant or at the silicone chain end,
groups having from 2 to 24 carbon atoms; phenylated silicones such
as phenyltrimethicones, phenyldimethicones,
phenyltrimethylsiloxydiphenylsiloxanes, diphenyldimethicones,
diphenylmethyldiphenyltrisiloxanes,
(2-phenylelthyl)trimethylsiloxysilica- tes, and
polymethylphenylsiloxanes;
[0152] mixtures thereof.
[0153] As emulsifiers and coemulsifiers which can be used in the
invention, there may be mentioned, for example, O/W emulsifiers
such as esters of a fatty acid and polyethylene glycol, in
particular PEG-100 stearate, and esters of the fatty acid and
glycerine such as glyceryl stearate, and W/O emulsifiers such as
oxyethylenated poly(methylcetyl)(dimethyl)methylsiloxane available
under the trademark ABIL WE09 from the company Degussa Goldschmidt
or the mixture of ethylene glycol acetyl stearate and glyceryl
tristearate marketed by the company Guardian under the trademark
UNITWIX.
[0154] As hydrophilic gelling agents, there may be mentioned in
particular carboxyvinyl polymers (carbomer), acrylic copolymers
such as acrylate/alkyl acrylate copolymers, polyacrylamides,
polysaccharides, natural gums and clays, and, as lipophilic gelling
agents, there may be mentioned modified clays such as bentones,
metal salts of fatty acids, hydrophobic silica and
polyethylenes.
[0155] As fillers which may be used in the composition of the
invention, there may be mentioned, for example, in addition to
pigments, silica powder; talc; starch crosslinked with
octenylsuccinic anhydride marketed by the company National Starch
under the name DRY FLO PLUS (28-1160); polyamide particles and in
particular those sold under the name ORGASOL by the company
Atochem; polyethylene powders; micropheres based on acrylic
copolymers, such as those in the form of an ethylene glycol
dimethacrylate/lauryl methacrylate copolymer sold by the company
Dow Corning under the name POLYTRAP; expanded powders such as
hollow microspheres in particular the microspheres marketed under
the name EXPANCEL by the company Kemanord Plast or under the name
MICROPEARL F 80 ED by the company Matsumoto; microbeads of silicone
resin such as those marketed under the name TOSPEARL by the company
Toshiba Silicone; and mixtures thereof. These fillers may be
present in quantities ranging from 0% to 20% by weight and
preferably from 1% to 10% by weight relative to the total weight of
the composition or of the preparation according to the
invention.
[0156] In order to further illustrate the present invention and the
advantages thereof, the following specific examples are given, it
being understood that same are intended only as illustrative and in
nowise limitative.
EXAMPLES
Example 1
[0157] Preparation of an Extract of Vitreoscilla filiformis
Containing Lipopolysaccharides:
[0158] (M. A. Apicella, J. McLeod Gritliss, and H. Schneider, 1994
Methods in enzymology, Vol. 235, (242-252))
[0159] Various methods make it possible to isolate the fraction of
interest containing the lipopolysaccharides:
[0160] the modified phenol-water method (Johnson and Perry, 1975,
Can J. Microbial, 22, p 29) described in paragraph 1;
[0161] the Darveau and Hancock method (1983, J. Bact. 155, p 831)
which uses SDS to solubilize the lipopolysaccharides, which makes
it possible to separate them from the insoluble peptidoglycan, this
method is described in paragraph 2. The proteins are removed from
the fraction containing the lipopolysaccharides by enzymatic
digestion (Pronase), the lipopolysaccharide fraction is then
precipitated with ethanol.
[0162] the method of extraction using proteinase K described in
paragraph 3.
[0163] 1. Modified Phenol Technique:
[0164] 1.1. Preparation of the Crude Lipopolysaccharides:
[0165] 25 ml of 50 mM Na phosphate buffer, pH 7, containing 5 mM
EDTA, are added to 5 g of Vitreoscilla filiformis bacteria (ATCC
15551) frozen or dried with acetone in powdered form, and the
mixture is stirred.
[0166] The following steps are then carried out:
[0167] adding 100 mg of lysozyme, stirring overnight at 4.degree.
C. and then incubating at 37.degree. C. for 20 min;
[0168] centrifuging for 3 min at low speed,
[0169] adjusting the volume to 100 ml with 50 mM Na phosphate
buffer pH 7 containing 20 mM MgCl.sub.2;
[0170] adding Rnase, Dnase (at 1 .mu.g/ml), incubating for 60 min
at 37.degree. C. and then for 60 min at 60.degree. C.;
[0171] the bacterial suspension is placed on a bath at 70.degree.
C., and an equal volume of 90% phenol (w/v), preheated to
70.degree. C., is added thereto,
[0172] it is cooled by stirring for 15 min on an ice bath,
[0173] centrifuging at 18,000 g for 15 min at 4.degree. C.
[0174] A marked interface is produced between the aqueous and
phenolic phases. The aqueous phase contains the
lipopolysaccharides; after dialyzing against water, this phase is
freeze-dried.
[0175] 1.2. Purification of the Crude Lipopolysaccharides
[0176] 20 to 35 mg of the lipopolysaccharides/ml of distilled water
are centrifuged at low speed (1100 g, 5 min). The supernatant
obtained is then centrifuged at high speed (105,000 g, 16 h,
4.degree. C.). The pellet is suspended in water, the centrifugation
is repeated until purified lipopolysaccharides are obtained. The
final pellet is resuspended in water and freeze-dried.
[0177] 2. SDS Method:
[0178] 15 ml of 10 mM Tris-HCl buffer pH 8 containing 2 mM
MgCl.sub.2, 100 .mu.g/ml Dnase and 25 .mu.g/ml Rnase are added to
500 mg of dried Vitreoscilla filiformis bacterial cells (ATCC
15551). The mixture is subjected to a French press twice, 15,000
psi, and then sonication, twice at 6 W, 30 sec.
[0179] The following steps are then applied:
[0180] adding Dnase 200 .mu.g final and Rnase 50 .mu.g final,
incubating 37.degree. C. 2 h,
[0181] adding 5 ml 0.5 M EDTA in 10 mM Tris-HCl pH 8, 2.5 ml 20%
SDS dissolved in 10 mM Tris-HCl and 2.5 10 mM Tris-HCl pH 8.
[0182] The final volume obtained is 25 ml containing 0.1 M EDTA; 2%
SDS and 10 mM Tris-HCl pH 9.5, the mixture is vortexed and
centrifuged at 50,000 g for 30 min at 20.degree. C.
[0183] The supernatant is separated after settling out. The
sediment which contains the peptidoglycan is discarded. Pronase is
added to the supernatant at a final concentration of 200 .mu.M,
incubation follows at 37.degree. C. overnight, with stirring (if a
precipitate forms, remove it by centrifuging at 1000 g 10 min).
[0184] The lipopolysaccharides are precipitated with 95% ethanol
(v/v) containing 0.376 M MgCl.sub.2 -70.degree. C., and then
centrifuged (12,000 g, 15 min, 4.degree. C.).
[0185] The pellet obtained is suspended in 25 ml 2% SDS, 0.1 M
EDTA, 10 mM Tris-HCl pH 8, sonicated and then incubated at
85.degree. C., 10 to 30 min. After cooling, the solution is
adjusted to pH 9.5.
[0186] Pronase is added at 25 .mu.g/ml, incubation at 37.degree.
C., overnight, with stirring.
[0187] The lipopolysaccharides are again precipitated with ethanol
95% (v/v) containing 0.376 M MgCl.sub.2, and then centrifuged
(12,000 g 15 min 4.degree. C.). To remove the insoluble
Mg.sup.2+-EDTA crystals, the pellet is resuspended in 15 ml of 10
mM Tris-HCl buffer pH 8, sonicated and centrifuged (1000 g, 5 min).
The supernatant is then centrifuged (200,000 g, 2 h 15.degree. C.)
in the presence of 25 mM MgCl.sub.2. The pellet which contains the
lipopolysaccharides is suspended in distilled water.
[0188] 3. Extraction of the Lipopolysaccharides Using Proteinase K
(Zanen and, 1988, FEMS Microbiology Letters 50, 85-88):
[0189] 30 ml of buffer containing 2% SDS, 5% 2-mercaptoethanol, 10%
glycerol and 0.25 M Tris-HCl pH 6.8 are added to 1.5 g of
freeze-dried Vitreoscilla filiformis cells (ATCC 15551).
[0190] The mixture is incubated for 15 to 30 min, 100.degree. C.,
centrifuged (10,000 g, 30 min, 4.degree. C.). The supernatant (20
ml) is recovered, 12 mg of proteinase K are added, the medium is
incubated at 60.degree. C. for 1 h and the lipopolysaccharides are
precipitated with 95% ethanol (v/v) containing 0.376 M MgCl.sub.2,
-20.degree. C. overnight, reprecipitating the lipopolysaccharides
with 95% ethanol (v/v) under the same conditions as above, the
pellet obtained is suspended in 10 ml of water, dialysed and then
freeze-dried.
Example 2
[0191] Formulations:
[0192] Composition 1--Regenerating Cream:
1 Extract of Example 1 (freeze-dried 0.1% extract obtained
according to method 3) Carbomer 940 .RTM. (cross-linked polyacrylic
acid) 0.3% Triethanolamine 0.3% Stearic acid 3.0% Cetyl alcohol
2.0% Self-emulsifiable glyceryl monostearate 3.0% Soyabean oil
10.0% Lanolin alcohol 2.0% Isopropyl myristate 4.0% Cetyl stearyl
2-ethylhexanoate 4.0% Perhydrosqualene 3.0% Paraffin 2.0% Glycerine
3.0% Preservatives 0.3% Water qs 100%
[0193] To prepare this cream, the aqueous phase containing the
glycerine, the preservatives and the water is heated to 80.degree.
C.; the Carbomer 940 is dispersed therein and then neutralized with
dry ethanolamine. The fatty phase, heated, and homogenized, to
80.degree. C., is introduced, with vigorous stirring, into the
aqueous phase. The extract of the example is dispersed in 10 g of
water and introduced at 40.degree. C. into the cream, with
stirring. The whole is cooled to room temperature. This cream is
applied to the skin of the face and of the neck once or twice by
day. It makes it possible in particular, after using for a few
days, to increase the regeneration of the epidermis and to give a
younger appearance to the skin.
[0194] Composition 2--Care Gel for the Face:
2 Extract of Example 1 (freeze-dried extract 0.05% obtained
according to method 3) Hydroxypropylcellulose (Klucel H sold by the
1.00% company Hercules) Antioxidant 0.05% Isopropanol 40.00%
Preservative 0.30% Water qs 100%
[0195] This gel is obtained by mixing the constituents in water and
adding the gelling agent last.
[0196] As for Composition 1, it may be applied twice by day; it is
particularly suitable for application in the morning because it
does not leave the skin greasy.
[0197] Composition 3--Regenerating Cream:
3 Extract of Example 1 (freeze-dried extract 1.0% obtained
according to method 3) Preservatives 0.85% Alcohol 5.0% Tocopheryl
acetate 1.0% Disodium EDTA 0.05% PEG-20 methyl glucose
sesquistearate 2.0% Glycerine 7.0% Acrylate polymer 0.25%
Cholesterol 0.1% Cyclohexasiloxane 3.5% Squalane 9.5% Water and
extract of Fagus sylvatica 2.0% Ceramide 0.05% Ammonium
polyacryloyldimethyl taurate 2.2% Hydrolysed lupin protein 1.0%
Vegetable oils 6.0% Polycaprolactone and Solanum lycopersicum 1.0%
(tomato) extract (lycopene) Divinyldimethicone/dimethicone
copolymer 2.0% (and) C12-13 pareth-3 (and) C12-13 pareth-23 Water
qs 100%
[0198] This cream is preferably intended to be applied daily in the
evening after cleansing the skin. This cream rapidly allows the
skin to be better moisturized and to be softer; it makes the
complexion bright and uniform and has a tightening effect which
softens wrinkles and fine lines and smoothens the skin.
[0199] Composition 4--Regenerating Day Fluid:
4 Extract of Example 1 (freeze-dried extract 1.0% obtained
according to method 3) Octyldodecanol 0.1% Preservatives 0.75%
Tocopheryl acetate 1.0% PEG-20 methyl glucose sesquistearate 3.0%
Sodium hyaluronate 0.1% Glycerine 7.0% Cyclohexasiloxane 9.5% Water
and extract of Fagus sylvatica 1.0% Ammonium polyacryloyldimethyl
taurate 1.0% Cyclopentasiloxane (and) dimethicone 7.5% Hydrolysed
lupin protein 1.0% Polycaprolactone and Solanum lycopersicum 1.0%
(tomato) extract (lycopene) Vegetable oils 3.0%
Divinyldimethicone/dimethicone copolymer 2.0% (and) C12-13 pareth-3
(and) C12-13 pareth-23 Water qs 100%
[0200] This fluid may be applied daily in the morning and in the
evening. As for the preceding cream, this fluid improves the visual
appearance of the skin and of the complexion by virtue of better
moisturizing and a tightening effect.
[0201] Composition 5--Care Cream for Solar Erythema (Oil-In-Water
Emulsion):
5 Extract of Example 1 (freeze-dried extract 0.75% obtained
according to method 3) Glyceryl stearate 2.00% Polysorbate 60
(Tween 60 sold by the company ICI) 1.00% Stearic acid 1.40%
Glycyrrhetinic acid 2.00% Triethanolamine 0.70% Carbomer 0.40%
Liquid fraction of shea butter 12.00% Sunflower oil 10.00% Lycopene
0.05% Perfume 0.50% Preservative 0.30% Water qs 100%
[0202] This cream should be applied twice by day for at least two
days, and then once by day until the erythema completely
disappears. By virtue of such an application, the skin more rapidly
rediscovers its normal appearance.
[0203] Composition 6--Sunscreen:
6 Extract of Example 1 (freeze-dried extract 3.5% obtained
according to method 3) Mixture of cetylstearyl alcohol and 7.0%
oxyethylenated cetylstearyl alcohol (33 EO) 80/20 Mixture of
glyceryl mono- and distearate 2.0% Cetyl alcohol 1.5%
Polydimethylsiloxane 1.5% Liquid paraffin 15.0% Butyl
methoxydibenzoylmethane 3.0% Octocrylene 7.0% Glycerine 20.0%
Demineralized water qs 100%
[0204] The application of this cream should precede exposure to the
sun and should be repeated every two hours. Such an application
prevents the damage which may be caused by the sun and accelerates
the repair of this possible damage.
[0205] Composition 7--Composition for Preparing the Skin for the
Sun:
[0206] (The ingredients are indicated under their CTFA names)
7 Extract of Example 1 (freeze-dried extract 0.5% obtained
according to method 3) Preservatives 1.35% Sodium citrate 0.035%
PEG-40 1.25% Pentaerythrityl tetraethylhexanoate 4% Glycerine 7%
Sorbitan tristearate 0.3% Prunus Armeniaca (apricot) Kernel oil 2%
Cetyl alcohol 0.7% Propylene glycol 2% Triethanolamine 0.4%
Cyclohexasiloxane 2% Carbomer 0.75% Tocopherol 1% Silica 2%
Ascorbyl glucoside 0.1% Water - titanium dioxide - silica - alumina
3% Polycaprolactone - betacarotene 5% Water qs 100%
[0207] Daily application of this composition at least one week
before solar exposure reduces the risks of erythema, burning and
oedema (in particular if the skin is protected with Composition 4
during solar exposure) and makes if possible to obtain a tanning of
better quality and longer lasting.
Example 3
[0208] Measurement of the Increase in the Synthesis of MnSOD:
[0209] I. Materials and Methods:
[0210] Cell Cultures and Treatment:
[0211] The studies are carried out on normal human fibroblasts and
on normal human epidermal keratinocytes in culture.
[0212] The dermal fibroblasts are cultured in DMEM medium (Life
Technology, ref. 21969035) containing L-glutamine (2 mM, Life
Technology, ref. 25030024), penicillin at 50 IU/ml and streptomycin
at 50 .mu.g/ml (Life Technology, ref. 15070063) and foetal calf
serum at 1% (v/v, Life Technology, ref. 10106151).
[0213] The keratinocytes are cultured in SFM medium with no
pituitary extracts and EGF.
[0214] The extract of Vitreoscilla filiformis obtained by applying
method 3 of Example 1 was applied to the cells at the concentration
of 0.1% (W/V). The contact time was 24 or 48 hours.
[0215] CDNA Array:
[0216] The methodology used is that recommended by Clontech (Palo
Alto, USA). The extraction/purification of total RNA of each
culture led to the isolation of quantities of total RNA of the
order of 100 to 150 .mu.g. The solutions of total RNA are treated
with DNAse I in order to remove any trace of contaminating DNA,
according to the supplier's recommendations. The quality of the RNA
was then checked on agarose gel and the RNA solutions were adjusted
to 1 .mu.g/ml.
[0217] The next step was the purification of the messenger RNAs
(mRNAs) by hybridization of the poly(A) ends of the mRNAs with
biotinylated and capture-selective oligo(dT) primers on
streptavidin beads, according to the Atlaspure (Clontech) protocol.
The .sup.33P-labelled DNA probes were prepared by reverse
transcription of the mRNAs attached to poly(dT) beads, with the aid
of a pool of primers specific for the sequences immobilized on the
"arrays", in the presence of (.alpha..sup.33P)-dATP. This step used
the reagents and the protocol recommended by Clontech. The labelled
probes were purified by chromatography on an exclusion column and
the quality and the equivalence of the labelled probes was
evaluated by liquid scintillation counting.
[0218] cDNA array membranes containing 1176 genes were pretreated
and then the cDNAs immobilized on each membrane were hybridized
(68.degree. C., overnight) with the corresponding labelled probes.
The filters were then extensively washed and placed in individual
plastic bags for analysis. The analysis was carried out by direct
quantification of the radioactivity of the spots with the aid of a
Cyclone phospholmager (Packard). The results are expressed in
relative expression units (RE, radioactivity of the spot
corresponding to each gene, corrected for the background noise and
the differences in intensity of labelling of the probes).
[0219] RT-Q-PCR:
[0220] The pairs of primer used in this study are Mn+ superoxide
dismutase 2 precursor (size of the amplified fragment: 259 bp) and
cytosolic superoxide dismutase 1 (size of the amplified fragment:
298 bp).
[0221] The total RNAs are extracted with the aid of Tri-Reagent
according to the protocol recommended by the supplier. It is
followed by another extraction with chloroform and precipitation
with isopropanol. Potentially contaminating traces of DNA are
removed by treating with the DNA-free system (Ambion). The reverse
transcriptase reaction is then carried out in the presence of
oligo(dT) primer and the enzyme Superscript II (Gibco). Each step
is followed by quantification, by fluorescence, of the cDNA
synthesized and adjustment of the concentrations to 150 ng/ml.
Another quantification of each cDNA, after final dilution, is
carried out before the PCR reaction.
[0222] The PCR reactions (polymerase chain reactions) were carried
out by quantitative PCR with the "Light Cycler" system (Roche
Molecular Systems Inc.) and according to the procedures recommended
by the supplier. The reaction mixture (10 .mu.l final) introduced
into the capillaries of a thermocycler is composed of 2.5 .mu.l of
cDNA, primers for the two markers, reaction mixture (Roche)
containing the enzyme Taq DNA polymerase, the marker SYBR Grenn I.
The PCR conditions are the following: activation 10 min at
95.degree. C., PCR reactions in 40 cycles, annealing 5 sec at
95.degree. C. and then 5 sec at 60.degree. C.
[0223] The analysis of fluorescence in the amplified DNA is
measured continuously during the PCR cycles. The mean value of the
relative expression (RE) is expressed as Arbitrary Units (AU)
calculated from the values of cycles of two independent PCRs
according to the following formula: (1/2.sup.number of
cycles).times.10.sup.6.
[0224] The results of expression of SOD are compared with that of
actin in order to take into account possible differences in cell
concentration in both cellular populations.
[0225] Flow Cytometry Study:
[0226] The cells are precultured at high density for 48 hours and
then treated or otherwise with the test product for 24 or 48 hours.
The cells are trypsinized and then rinsed with a PBS/2% FCS
solution. The cells are transferred to Eppendorf tubes and
centrifuged for 5' at 1500 rpm. The cells are fixed with a
PBS-formalin solution at 4% final, 30' at room temperature and in
the dark. They are then permeabilized with the aid of a 0.1%
Triton-X100/0.1% citrate solution. The cells are labelled in the
presence of anti-MnSOD (TEBU SOD-110) and anti-Cu/Zn SOD (TEBU
SOD-100) antibodies according to the supplier's instructions.
Secondary labelling is carried out with the aid of secondary
antibodies/FITC (TEBU L 42001) according to the supplier's
instructions. Analysis of the samples is carried out by cytometry
(FACSCAN cytometry, Cell Quest software; Becton-Dickinson) on the
total population. The statistics are performed on 10,000 cells of
each sample. The results are expressed as intensity of fluorescence
(IF) corresponding to the relative quantity of each marker per cell
in a total population of 10,000 cells analyzed.
[0227] II. Results:
[0228] 2.1) Study of cDNA Macroarray:
[0229] 2.1.1) On Culture of Human Keratinocytes:
8 Comparison of control Population population and population
Control exposed to exposed to the extract Genes population the
extract (in %) Mn + SOD2 1.3 3.3 244 precursor (SOD2) Cytosolic
SOD1 4.9 6.8 138 (SOD1)
[0230] It is observed that the cell population consisting of
keratinocytes which is exposed to the extracts of a nonfruiting
nonphotosynthetic filamentous bacterium contains significantly more
mRNA for SOD2 than the control population whereas the quantity of
mRNA for SOD1 is only slightly increased.
[0231] 2.1.2) On Culture of Human Fibroblasts:
9 Comparison of control Population population and population
Control exposed to exposed to the extract Gene population the
extract (in %) Mn + SOD2 1.9 9.3 481 precursor (SOD2)
[0232] Just as for the keratinocytes, a cell population consisting
of fibroblasts exposed to an extract of a nonfruiting
nonphotosynthetic filamentous bacterium contains nearly 5 times
more niRNA for SOD2 than the control population.
[0233] 2.2) PCR Study:
[0234] The results below are those obtained on a culture of human
keratinocytes.
[0235] 2.2.1) Results on Cytosolic Superoxide Dismutase 1
(SOD1):
10 RE* RE* Actin SOD1 Actin SOD1 SOD1/ % Treatment Cycles Cycles
(AU) (AU) Actin Control Control 16.12 20.12 13.99 0.93 6.64 100
16.13 19.96 10-2 VF 16.23 20.02 12.88 0.94 7.28 110 Extract 16.26
20.03 10-2 (VF: Vitreoscilla filiformis)
[0236] These results confirm those observed in point 2.1.1., that
is to say that the exposure of a population of keratinocytes to the
bacterial extract appeared according to method 3 does not cause
transcriptional induction of SOD1.
[0237] 2.2.2) Results on Mn+ Superoxide Dismutase (SOD2):
11 RE* RE* Actin SOD2 Actin SOD2 SOD2/ % Treatment Cycles Cycles
(AU) (AU) Actin Control Control 20.78 26.45 0.55 0.01 2.37 100
20.80 25.97 10-2 VF 20.79 23.65 0.51 0.09 1.79 755 Extract 21.02
23.16 10-2 (VF: Vitreoscilla filiformis)
[0238] These results also confirm those of point 2.1.1.: the
quantity of mRNA encoding SOD2 is much higher in the cell
population which was exposed to the bacterial extract.
[0239] 2.3) Flow Cytometry Study:
[0240] The results below are those obtained on a culture of human
keratinocytes.
[0241] 2.3.1) Results on Cytosolic Superoxide Dismutase 1
(SOD1):
[0242] Relative quantity of Cu/Zn SOD in control keratinocytes and
in keratinocytes treated with the extract (0.1%, W/V) for 24
hours.
12 Number of Treatment IF Standard Deviation Sample % p Control
180.9 15.3 3 100 -- VF 217.8 7.3 3 120 p < 0.01 Extract (VF:
Vitreoscilla filiformis)
[0243] It is observed that although exposure to the VF extract does
not induce transcription of mRNA for SOD1, its expression is
statistically increased (+20%).
[0244] 2.3.2) Results on Mn Superoxide Dismutase (SOD2):
[0245] Relative quantity of MnSOD in control keratinocytes and
keratinocytes treated with the extract (0.1%) for 48 hours.
13 Number of Treatment IF Standard Deviation Sample % p Control
20.2 2.6 3 100 -- VF 28.1 1.8 3 139 p < 0.01 Extract (VF:
Vitreoscilla filiformis)
[0246] These trials also show an increase in the cellular
concentration of SOD2 (+39%) after exposure to the VF extract.
[0247] The observation of these trials leads to the conclusion that
the fibroblasts exposed in the presence of the extract obtained in
Example 1 highly express the messenger RNAs encoding MnSOD (2.1.
and 2.2.) and this induction could be confirmed by observing an
increase in the protein presence of MnSOD (2.3.), in particular in
the mitochondria (visualized by microscopy observations).
[0248] Each patent, patent application and literature
article/report cited or indicated herein is hereby expressly
incorporated by reference.
[0249] While the invention has been described in terms of various
specific and preferred embodiments, the skilled artisan will
appreciate that various modifications, substitutions, omissions,
and changes may be made without departing from the spirit thereof.
Accordingly, it is intended that the scope of the present invention
be limited solely by the scope of the following claims, including
equivalents thereof.
* * * * *