U.S. patent application number 13/796457 was filed with the patent office on 2013-07-25 for polysaccharides compositions comprising fucans and galactans and their use to reduce extravasation and inflammation.
This patent application is currently assigned to LUCAS MEYER COSMETICS CANADA INC.. The applicant listed for this patent is Lucas Meyer Cosmetics Canada Inc.. Invention is credited to PATRICE DIONNE, ALAIN LAVOIE, JEAN-YVES MOIGNE, ALAIN THIBODEAU.
Application Number | 20130190269 13/796457 |
Document ID | / |
Family ID | 37836181 |
Filed Date | 2013-07-25 |
United States Patent
Application |
20130190269 |
Kind Code |
A1 |
THIBODEAU; ALAIN ; et
al. |
July 25, 2013 |
POLYSACCHARIDES COMPOSITIONS COMPRISING FUCANS AND GALACTANS AND
THEIR USE TO REDUCE EXTRAVASATION AND INFLAMMATION
Abstract
A method for inhibiting the release of one or more of IL-8, PGE2
and VEGF by a cell activated during an inflammatory process, said
method comprising administering to said cell an anti-inflammatory
polysaccharide composition comprising fucans and galactans to a
cell, wherein the ratio of fucans to galactans in said composition
enables a reduction in the cytotoxicity of said galactans and an
anti-inflammatory composition comprising a ratio of brown algae
fucans/red algae galactans of ranging from about 2.5/1 (w/w) to
about 40/1 (w/w), the galactans having a molecular weight higher
than about 100 kDa, and the fucans having a molecular weight
ranging from about 0.1 kDa and 100 kDa.
Inventors: |
THIBODEAU; ALAIN;
(ST-AUGUSTIN-DE-DESMAURES, CA) ; LAVOIE; ALAIN;
(SAINTE-FOY, CA) ; DIONNE; PATRICE;
(ST-REDEMPTEUR, CA) ; MOIGNE; JEAN-YVES;
(OUESSANT, FR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Lucas Meyer Cosmetics Canada Inc.; |
Quebec |
|
CA |
|
|
Assignee: |
LUCAS MEYER COSMETICS CANADA
INC.
QUEBEC
CA
|
Family ID: |
37836181 |
Appl. No.: |
13/796457 |
Filed: |
March 12, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11992182 |
Apr 20, 2009 |
8426381 |
|
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PCT/CA2006/001496 |
Sep 11, 2006 |
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13796457 |
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60762488 |
Jan 27, 2006 |
|
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60715178 |
Sep 9, 2005 |
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Current U.S.
Class: |
514/54 ;
435/375 |
Current CPC
Class: |
A61K 36/03 20130101;
A61K 36/04 20130101; A61K 31/715 20130101; A61K 36/04 20130101;
A61P 29/00 20180101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 36/03 20130101 |
Class at
Publication: |
514/54 ;
435/375 |
International
Class: |
A61K 31/715 20060101
A61K031/715 |
Claims
1. A method for inhibiting the release of one or more of IL-8, PGE2
and VEGF by a cell activated during an inflammatory process, said
method comprising administering to said cell an anti-inflammatory
polysaccharide composition comprising fucans and galactans to a
cell, wherein the ratio of fucans to galactans in said composition
enables a reduction in the cytotoxicity of said galactans.
2. The method of claim 1, wherein the release of IL-8, PGE2 and
VEGF is from epithelial cells.
3. The method of claim 1, wherein the fucans increase the
inhibition of inflammation.
4. The method of claim 1, wherein (a) the galactans comprise
galactans obtained from red algae; and/or (b) the fucans comprise
fucans obtained from brown algae.
5. The method of claim 4, wherein (a) the red algae comprises
Asparagopsis armata; and/or (b) the brown algae comprises
Ascophyllum nodosum.
6. The method of claim 1, wherein the galactans have an average
molecular weight higher than about 100 kDa.
7. The method of claim 1, wherein the galactans have an average
molecular weight of about 350 kDa.
8. The method of claim 1, wherein (a) the galactans have a
galactose content of about 30% to about 40% of dry weight of the
galactans; (b) the galactans have an uronic acid content of about
1% to about 5% of dry weight of the galactans; (c) wherein the
galactans have a sulfate content of about 20% to about 35% of dry
weight of the galactans; (d) the fucans have an average molecular
weight ranging from about 0.1 kDa to about 100 kDa; (e) the fucans
have a fucose content of about 20-35% of dry weight of the fucans;
(f) the fucans have an uronic acid content of about 10 to about 29%
of dry weight of the fucans; (g) the fucans have a sulfate content
of about 15 to about 25% of dry weight of the fucans; (h) the
fucans anf the galactans in said composition are present in a ratio
of from about 2.5 fucans/1 galactans (w/w) to about 40 fucans/1
galactans (w/w); (i) the galactans comprise native galactans; (j)
the fucans comprise native fucans; (k) the fucans comprise
depolymerized fucans; (l) the fucans comprise demineralized fucans;
and/or (m) the galactans comprise demineralized galactans.
9. The method of claim 8, wherein (a) the galactans have a
galactose content of about 37% of dry weight of the galactans; (b)
the galactans have an uronic acid content of about 3% of dry weight
of the galactans; (c) the galactans have a sulfate content of about
27% of dry weight of the galactans; (d) the fucans have an average
molecular weight ranging from about 5 kDa to about 25 kDa; and/or
(e) the fucans and the galactans are present in a ratio of about 10
fucans/1 galactans (w/w) in the composition.
10. The method of claim 1, wherein said composition is administered
topically.
11. The method of claim 10, for (a) alleviating or improving skin
disorders or conditions caused by UV exposure, chemical stress or
aggression from pollutants, exfoliating agents or skin irritants;
or (b) preventing skin disorders or conditions caused by UV
exposure, chemical stress or aggression from pollutants,
exfoliating agents or skin irritants.
12. The method of claim 1, wherein the cell is within a
subject.
13. The method of claim 12, wherein the polysaccharide composition
is topically administered to the subject.
14. The method of claim 13, wherein the subject is a human affected
by skin disorders or conditions caused by UV exposure, chemical
stress, or aggression from pollutants, exfoliating agents or skin
irritants.
15. An anti-inflammatory composition comprising brown algae fucans
and red algae galactans, wherein the ratio of brown algae
fucans/red algae galactans in said composition is ranging from
about 2.5/1 (w/w) to about 40/1 (w/w), said galactans having a
molecular weight higher than about 100 kDa and said fucans having a
molecular weight ranging from about 0.1 kDa and 100 kDa, wherein
the ratio of fucans to galactans in said composition enables a
reduction in the cytotoxicity of said galactans.
16. The composition of claim 15, wherein (a) the galactans have an
average molecular weight of about 350 kDa; (b) the galactans have a
galactose content of about 30% to about 40% of dry weight of the
galactans; (c) the galactans have an uronic acid content of about
1% to about 5% of dry weight of the galactans; (d) the galactans
have a sulfate content of about 20% to about 35% of dry weight of
the galactans; (e) the fucans have an average molecular weight
ranging from about 5 kDa to about 25 kDa; (f) the fucans have a
fucose content of about 20-35% of dry weight of the fucans; (g) the
fucans have an uronic acid content of about 10 to about 29% of dry
weight of the fucans; (h) the fucans have a sulfate content of
about 15 to about 25% of dry weight of the fucans; (i) the ratio of
brown algae fucans/red algae galactans is of 10/1; (j) the brown
algae is Ascophyllum nodosum; (k) the red algae is Asparagopsis
armata; (l) the galactans comprise native galactans; (m) the fucans
comprise native fucans; (n) the fucans comprise depolymerized
fucans; (o) the fucans comprise demineralized fucans; and/or (p)
the galactans comprise demineralized galactans.
17. The composition of claim 16, wherein (a) the galactans have a
galactose content of about 37% of dry weight of the galactans; (b)
the galactans have an uronic acid content of about 3% of dry weight
of the galactans; and/or (c) the galactans have a sulfate content
of about 27% of dry weight of the galactans.
18. The composition of claim 15, wherein the galactans have a
molecular weight of about 350 kDa, and the fucans have a molecular
weight ranging from about 15 kDa and 25 kDa.
19. The composition of claim 15, which is a topical
composition.
20. The composition of claim 15, which is a cosmetic composition.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of U.S.
patent application Ser. No. 11,992,182 filed on Apr. 20, 2009 now
pending, which is a National Entry Application of PCT application
no PCT/CA2006/001496 filed on 11 Sep. 2006 and published in English
under PCT Article 21(2), which itself claims benefit of U.S.
provisional application Ser. No. 60/715,178, filed on 9 Sep. 2005
and on U.S. provisional application Ser. No. 60/762,488, filed on
27 Jan. 2006. All documents above are incorporated herein in their
entirety by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to polysaccharides
compositions comprising fucans and galactans and their use to
reduce extravasation and inflammation. More specifically, the
present invention is concerned with use of polysaccharide
compositions comprising fucans and galactans obtained from brown
and red algae, respectively, which have anti-inflammatory and
vascular protective activities. The invention also relates to a
method for treating, preventing or alleviating the symptoms of
disorders and diseases associated with excess levels of
pro-inflammatory mediators, extravasation stimulating agents and
consequent vascular and extracellular effects. Amongst these
diseases or disorders are those affecting skin, caused by UV,
pollutants exposure, stress, psoriasis, acne, rosacea, skin cancer,
and/or skin aging.
BACKGROUND OF THE INVENTION
[0003] Fucans are polysaccharides originating mainly from the cell
walls of shoots of brown algae (Pheophyceae family) belonging to
the Ascophyllum, Fucus, Pelvetia and Himmanthali genera. They are
also found in some marine animals, such as sea urchins and sea
cucumbers. Fucansobtained by extraction from the cell walls of
brown algae shoots, also termed fucoidans when in their sulfated
form, consist of a heterogeneous population of molecules which
comprises principally sulfated L-fucose polymers of average molar
mass ranging from 5000 to 800,000 g/mol. These polymers also
contain uronic acids. Whilst sulfatation degree, molecular weight,
and structure of sugar residues of fucans vary among species,
several studies clearly show that brown algae fucans, for example,
Ascophyllum nodosum fucans possess a large portion of both
.alpha.(1.fwdarw.3) and .alpha.(1.fwdarw.4) glycosidic bonds.
[0004] Fucans have varied biological activities: it was shown that
they have anticoagulant and antithrombotic activities (T. Nishino
and T. Nagumo, Carbohydr. Res. 229, p. 355-362, (1992); Application
EP 0403 377; S. Colliec et al. Thromb. Res. 64, p. 143-154 (1991);
S. Soeda et al. Thromb. Res. 72, p. 247-256 (1993); Mauray et al.
Thromb. Haemost. (5) 1280-1285 (1995)), they can protect cells
against viral infection (M. Baba et al. J. AIDS, 3, p. 493-499,
(1990)), they have antiangiogenic (R. Hahnenberger and A. M.
Jackobson, Glycoconjugate J., 8, 350-353 (1991)) and
anticomplementary (C. Blondin et al., Mol. Immunol., 31, p.
247-253, (1994)) activities. It has also been observed that they
can act as modulators of cell adhesion (C. G. Glabe et al., J. Cell
Sci., 61, p. 475-490, (1983)), of growth factor release (D. A.
Belfort et al., J. Cell. Physiol. 157, p. 184-189, (1993)), of
tumor cell's (M. Ellouali et al., Anticancer Res., 13, p. 2011-2020
(1993); D. R. Coombe et al., Int. J. Cancer, 39, pp. 82-90, (1987);
D. Riou et al., Anticancer Res., 16, 1213-1218 (1996)) and of
vascular smooth muscle cell's proliferation (Logeart et al., Eur.
J. Cell. Biol., 74, pp. 376-384 (1997)), and can block
spermatozoid/ovule interactions in various species (M. C. Mahony et
al., Contraception, 48, p. 277-289, (1993)).
[0005] Galactans are other polysaccharides originating mainly from
the cell walls of red algae (Redphyceae family). The most abundant
galactans found in the red algae are carrageenans and agarans.
These polysaccharides play a significant physiological role in the
resistance of mechanical stress, hydration, and in both the ionic
and the osmotic regulation required within marine environments. Raw
galactans are obtained by extraction from the cell walls of red
algae shoots, and consist of a heterogeneous population of
molecules which comprises mainly sulfated beta-D-galactose, xylose
and galactose polymers of average molar mass ranging from 50,000 to
800,000 g/mol.
[0006] Galactans have varied known biological activities: They have
anticoagulant (Melo et al., J. Biol. Chem. 279:20824-35(2004);
Pereira et al., 1999; Facia et al., J. Biol. Chem. 275:29299-307
(2000)) and antiviral activities (Duarte et al., Carbohydr Res.
339:335-47(2004); Huleihel et al., Appl Spectrosc. 57:390-5.
(2003); Carlucci et al., Planta Med. 63:429-32 (1997)). It has also
been observed that they can act as modulators of proliferation of
tumor cells (Zhou et al., Pharmacol. Res. 50:47-53 (2004); Geresh
et al J Biochem Biophys Methods. 50:179-87 (2002)).
[0007] Processes for obtaining fucans and galactans from a
plurality of species have been summarized in Tables 1 and 2,
respectively. Generally, brown algae are good sources of fucans
while red algae are good sources of galactans.
TABLE-US-00001 TABLE 1 BROWN ALGAE AS SOURCES OF SULFATED-FUCANS
AND KNOWN PROCESSES FOR THEIR OBTENTION Species References
Ascophyllum nodosum Marais and Joseleau (Carbohydr. Res 336:
155-159; 2001), Mabeau et al. Phytochemistry 29: 2441-2445; 1990),
Pereira et al. (J. Biol. Chem 274(12): 7656-7667; 1999), Tissot et
al. (Biochim. Biophys. Acta 165(1-2): 5-16; 2003) Fucus sp.
Chevolot et al. (Carbohydr. Res. 330(4): 529-535; 2001), Bilan et
al. (Carbohydr. Res. 337(8): 719-730; 2002)), Bilan et al.
(Carbohydr. Res. 339(3): 511-517; 2004), Ruperez et al. (J. Agric.
Food Chem. 50(4): 840-845; 2002 Stichopus japonicus Kariya et al.
(Carbohydr. Res. 339(7): 1339-1346; 2004) Sargassum sp. Duarte et
al. (Carbohydr. Res. 333(4): 282-293; 2001), Zhu et al. (Biochem.
Cell Biol. 81(1): 25-33; 2003), Zhuang et al. (Biosci. Biotechnol.
Biochem. 59(4): 563-567; 1995), Nagaoka et al. (Glycoconj. J. 16:
19-26; 1999) Padina gymnospora Andrade et al. (J. Struct. Biol.
145(3): 216-225; 2004) Adenocystis utricularis Ponce et al.
(carbohydr. Res 338(2): 153-165: 2003) Cladosiphon okamuranus Sakai
et al. (Mar. Biotechnol. 5(6): 536-544; 2003), Nagaoka et al.
(Glycoconj. J. 16: 19-26; 1999) Kjellmaniella crassifolia Sakai et
al. (Mar. Biotechnol. 4(4): 399-405; 2002), Nagaoka et al.
(Glycoconj. J. 16: 19-26; 1999) Pelvetia canaliculata Colliec etal.
(Phytochemistry 35: 697-700; 1991) Ecklonia kurome Nishino et al.
(Carbohydr. Res. 211(1): 77-90; 1991) Chorda filum Chizhov et al.
(Carbohydr. Res. 320(1-2): 108-119; 1999) Undaria pinnatifida Lee
etal. (Chem. Pharm. Bull. 52(9): 1091-1094; 2004 Laminaria japonica
Zvyagiintseva et al. (Comp. Bichem. Physiol. C. Toxicol. Pharmacol.
126(3): 209-215; 2000)
TABLE-US-00002 TABLE 2 RED ALGAE AS SOURCES OF GALACTANS AND KNOWN
PROCESSES FOR THEIR OBTENTION Species references Asparagopsis sp.
Haslin et al. (Planta Med. 67(4): 301-305; 2001) Bostrychia
montagnei Duarte et al. (Phytomedicine 8(1): 53-58; 2001) Corallina
sp. Usov et al. (Carbohydr. Res. 303(1): 93-102; 1997), Cases et
al. (Int. J. Biol. Macromol. 16(2): 93-97; 1994) Polysiphonia
lanosa Batey and Survey (Carbohydr. Res. 43(1): 133-43; 1975)
Gracilaria sp. Marinho-Soriano and Bourret (Bioresour. Technol.
96(3): 379-382; 2005), Freile-Pelegrin and Murano (Bioresour.
Technol. 96(3): 295-302; 2005) Acanthophora spicifer Duarte et al.
(Carbohydr. Res. 339(2): 335-347; 2004) Georgiella confluens
Kolander and Matulewicz (Carbohydr. Res. 337(1): 57-68; 2002)
Laurencia coronus Usov et Elashvili (Bioorg. Khim 23(6): 505-511;
1997) Porphyra haitanensis Zhang et al. (Carbohydr. Res. 339(1):
105-111; 2004) Botryocladia occidentalis Farias et al. (Thromb.
Haemost. 86(6): 1540-1546; 2001), Farias et al. (J. Biol. Chem.
275(38): 29299-29307; 2000) Cryptopleura ramosa Carlucci et al.
(Planta Med. 63(5): 429-432; 1997) Chondrus ocellatus Zhou et al.
(Pharmacol. Res. 50(1): 47-53; 2004) Gymnogongrus tolulosus Estevez
et al. (Carbohydr. Res. 331(1): 27-41; 2001) Phacelocarpus
peperocarpos Liao et al. (Carbohydr. Res; 296: 237-247; 1996)
[0008] Human epithelium plays an essential role in the equilibrium
and repair of connective tissues. It is in particular responsible
for renewing extracellular matrix (ECM), and in return its
functions are modified by the substances present in this
matrix.
[0009] In particular, in the process of tissue remodeling and
healing which intervene after an injury, the connective tissue is
the context for constant exchanges between all the cells involved
in this process. These exchanges take place in particular via
cytokines or soluble mediators which are transmitted through the
ECM.
[0010] For example, in the covering connective tissues such as the
cutaneous tissues, the healing process begins after the formation
of a provisional matrix (red thrombus), with the recruitment of
inflammatory cells (leukocytes, macrophages and polymorphonuclear
cells), which initiate a phase of destruction of the lesioned
tissue.
[0011] These inflammatory cells participate in the destruction by
secreting matrix proteinases such as collagenase (MMP8), leukocytic
or neutrophil elastase or cathepsin G, by liberating cytokines, and
in particular interleukin-1 (IL-1), which stimulate the
proliferation and migration of fibroblasts and of epithelial cells,
and the expression, by these cells, of certain metalloproteinases
such as interstitial collagenase (MMP1) or gelatinase B (MMP9).
[0012] This destruction phase, which begins very soon after the
injury, ends when the epithelium and its basement membrane have
been reconstituted.
[0013] It is followed by repair and resolution phases in which the
fibroblasts reconstruct and reorganize the collagen framework; the
expression by the fibroblasts of gelatinase A (MMP2) is in
particular observed, matrix metalloproteinase actively
participating in all the tissue remodeling phenomena.
[0014] Repercussion of UV Exposure on the Skin Microcapillary
Integrity:
[0015] The cutaneous layer is the primary external barrier
protecting the body from harm as a result of invading foreign
particles. In order to adequately perform its function, the skin
has been endowed with a range of endogenous surveillance systems.
Following tissue damage, free radicals are released with active
cytokines to counteract these non-self particles. Subsequently,
enzymatic activities proceed to dismantle damaged components such
as cell bodies and fibrillar components of the ECM. This occurs
before repair processes establish and are able to promote cellular
proliferation and biosynthetic activity of the ECM's
components.
[0016] At the microscopic level, the skin may be viewed as a highly
complex arrangement of diversified cell types which are embedded
within the ECM. In addition to serving as a structural scaffold,
the ECM functions as a highway providing the means for cell
movement, migration and differentiation. It also functions as a
signal transduction pathway through which chemical mediators are
able to travel between individual cells and the superposed skin
layers. To a certain extent, the skin's ECM may be conceptualized
as a loose interlaced cotton weave into which cells are nested and
able to interact with one another and their surrounding
environment.
[0017] The ECM appears as a complex array of macromolecules and
fibrillar components. These components are fashioned with various
types of collagen fibers, elastin fibers, glycosaminoglycans and
glycoproteins. The ECM is both produced and organized by its
resident cells; mainly the keratinocytes and fibroblasts. This
amalgamated connective tissue is responsible for the firmness,
elasticity as well as the overall integrity of the skin. Despite
its highly intricate fibrillar composition, the ECM remains a
dynamic structure. As such, it must be involved in morphogenesis
and tissue repair, thus supporting cell proliferation and
macromolecular remodelling. The plasticity of the ECM can further
be demonstrated in its role in sensing external mechanical
forces.
[0018] It has been suggested that the application of tensile,
gravitational force and stretching forces to the skin trigger a
mechanochemical signal transduction (mechanosensing) involving the
direct ECM-cell and/or cell-cell interactions (Silver F H, Siperko
L M. Crit Rev Biomed Eng. 2003; 31(4):255-331). Specifically, the
ECM network acts as a sensor that informs skin cells on how to
adapt to dynamic environmental conditions. Through downstream
signal transduction, the skin's ECM may also influence other
tissues through their response to external stimuli (Eckes B, Krieg
T. Clin Exp Rheumatol. 2004 January-February; 22(3 Suppl
33):573-6). For instance, the mechanical forces imposed by a
tridimensional collagen network switch on "mechanical-responsive
genes" that favour a synthetic phenotype (Kessler D, Dethlefsen S,
Haase I, Plomann M, Hirche F, Krieg T, Eckes B. J Biol Chem. 2001
Sep. 28; 276(39):36575-85). This illustrates the ability of the ECM
to support both a biochemical role and an obvious physical function
as a home for resident cells. As a result of its intimate
interaction with the external environment, the ECM and its
associated structures are vulnerable to the continuous barrage of
external insults. Ultimately, the repeated effects of these insults
may affect the skin's health and appearance.
[0019] Chronological and actinic aging of the skin: Aging is a
multifactorial phenomenon. The aging of the skin is mainly the
result of one's genetic predisposition (known as chronological
aging) and one's physiological reaction to environmental stresses
(known as actinic aging). Chronological aging is largely
genetically driven and appears to be mainly a reduction in
anti-oxidant production (Finkel T, Holbrook N. J. Nature. 2000 Nov.
9; (408):239-247), cellular senescence and a general lowering of
anabolic activities (Jenkins G. Mech Ageing Dev. 2002 April;
123(7):801-10). Actinic aging seems to be skin specific and is
defined as the effect of the external environment on the skin's
biological response. The skin response to actinic aging, also
referred to as photo-damage, is typically associated with a lack of
normal hydration, apparition of telangiectasia, sagging of the skin
and the appearance of fine line and wrinkles.
[0020] Environmental insults, such as UV and/or polluting
deleterious chemicals found in the atmosphere are typically
encountered by keratinocytes of the epidermis which are located at
the outmost peripheral level of the skin. The reaction of
keratinocytes to the environmental stimuli triggers a cascade of
reactions where the acquisition of the initial signal is passed on
from cell to cell through a mechanism of biochemical
interpretations. As a result, the ensuing biological response may
be amplified and propagated to other layers of the skin. This
cascade of biochemical reactions, especially upon UV exposure, is
directly correlated to a number of histological damages that
accumulate to provoke the appearance of signs of actinic aging.
[0021] UV irradiation has been shown to have pleiotropic effects at
the skin level causing DNA lesions, cellular apoptosis,
immunosuppression and inflammation/erythema (Soter N A. Semin
Dermatol. 1990 March; 9(1):11-5, Matsumura Y, Ananthaswamy H N.
Expert Rev Mol Med. 2002 Dec. 2; 2002:1-22). With respect to
actinic skin aging--or photo-damage--the increase in matrix
metalloproteinase (MMP) activation and expression is the most
recognized degradation pathway induced as a result of the skin
exposure to UV (Fisher G J. CUTIS. 2005 February; (75):5-9). It has
been suggested that the proteolytic action of MMP causes the
breakdown of collagen fibers in the ECM and that the histological
damages which ensues eventually leads to the appearance of a
photo-damaged phenotype (Fisher G J, Wang Z Q, Datta S C, Varani J,
Kang S, Voorhees J J. New-England Journal of Medicine. 1997;
337:1419-1428; Fisher G J, Kang S, Varani J, Bata-Csorgo Z, Wan Y,
Datta S, Voorhees J J. Arch Dermatol. 2002 November;
138(11):1462-70; Brennan M, Bhatti H, Nerusu K C, Bhagavathula N,
Kang S, Fisher G J, Varani J, Voorhees J J. Matrix Photochemistry
and Photobiology. 2003; 78(1):43-48). Furthermore, it is suggested
that the reduction in the ability of fibroblasts to synthesize
collagen is reduced in photo-damaged skin as a result of a
decreased cell-ECM mechanical tension (Varani J, Schuger L, Dame M
K, Leonard C, Fligiel S E, Kang S, Fisher G J, Voorghees J J. J
Invest Dermatol. 2004 June; 122(6):1471-9).
[0022] Alteration to the tridimensional organization of the skin's
ECM is undoubtedly among the most significant and apparent
molecular changes during actinic aging. This microscopic
modification ultimately results in the macroscopic appearance of
cutaneous aging. Those changes triggered during actinic aging
involve the release of biochemical mediators that have pleiotropic
actions in skin structures. It is known that the phenomena of
microcapillary dilation and increased permeability figure among the
early cutaneous responses upon UV exposure.
[0023] Roles of VEGF and PGE.sub.2 in the conformational changes of
skin microcapillaries: The reactions of skin microcapillary
dilation and hyperpermeability are characteristic of an excessive
UV exposure and lead to the inflammation of the skin (Matsumura Y,
Ananthaswamy H N. Toxicol Appl Pharmacol. 2004 Mar. 15;
195(3):298-308). Microcapillary integrity is influenced by
biochemical mediators such as cytokines and other specific growth
factors found in the skin. Among these, the pro-inflammatory
prostaglandin E.sub.2 (PGE.sub.2) plays a central role in the
skin's response to stress as its expression is readily triggered by
inflammatory stimuli (Kabashima K, Miyachi Y. Journal of
Dermatological Science. 2004; 34:177-184; Lee J L, Mukhtar H,
Bickers D R, Kopelovich L, Altar M. Tox Appli Pharmacol. 2003;
(192):294-306; Bachelor M A, Bowden G T. Seminars in Cancer
Biology. 2004; 14:131-138). The expression of PGE.sub.2 is
upregulated in skin following exposure to UV (Hruza L L, Pentland A
P. J Invest Dermatol. 1993 January; 100(1):35S-41S) whereby it acts
upon specific cell receptors and mediates microcapillary dilation
(Lee J L, Mukhtar H, Bickers D R, Kopelovich L, Altar M. Tox Appli
Pharmacol. 2003; (192):294-306). The UV-induced increase in
PGE.sub.2 may be achieved through multiple signal transduction
pathways (Ashida M, Bito T, Budiyanto A, Ichihashi M, Ueda M.
Experimental Dermatology. 2003; 12:445-452). Furthermore, dilated
microcapillaries are also more susceptible to the leakage of
leukocytes into the ECM where they can release pro-inflammatory
cytokines, growth factors and degradation enzymes. This can be
demonstrated in the inflammatory skin condition rosacea, which is
characterized by the presence of dilated vessels (Van Zuuren E,
Graber M, Hollis S, Chaudhry M, Gupta A, Gover M. Cochrane Database
Syst Rev. 2005 Jul. 20; (3):CD003262) and a collapsed ECM structure
(Crawford G H, Pelle M T, James W D. J AM Acad Dermatol. 2004
September; 51(3):327-41).
[0024] The Vascular Endothelial Growth Factor (VEGF) is also a
factor known to be induced in keratinocytes and fibroblasts by a
specific range of effectors such as tissue hypoxia (Detmar M, Brown
L F, Berse B, Jackman R W, Elicker B M, Dvorak H F, Claffey K P. J
Invest Dermatol. 1997 March; 108(3):263-8.), pro-inflammatory
cytokines (Trompezinski S, Berthier-Vergnes O, Denis A, Schmitt D,
Viac J. Exp Dermatol. 2004 February; 13(2):98-105), nitric oxide
(Frank et al., 1999), toxins (Deasi A, Lankford H A, Warren J S.
Inflammation. 2000 February; 24(1):1-9) and upon exposure to UV.
VEGF is a regulator of angiogenesis (the formation of new blood
vessels) in inflammatory conditions (Detmar M, Brown L F, Schon M
P, Elicker B M, Velasco P, Richard L, Fukurama D, Monsky W, Claffey
K P, Jain R K. J Invest Dermatol. 1998 July; 111(1):1-6). For
instance, the expression of this growth factor has been shown to be
upregulated in psoriasis (Detmar, 1994) as well as in rosacea
(Lachgar S, Charveron M, Gall Y, Bonafe J L. Dermatology. 1999; 199
Suppl 1:25-7). Keratinocytes represent an important source of VEGF
(Ballaun C, Weninger W, Uthman A, Weich H, Tschachler. J Invest
Dermatol. 1995 January; 104(1):7-10). VEGF expression in these
cells may be induced via both UVA and UVB and it has been suggested
that this induction mechanism differs according to the specific
type of stimuli (Gille J, Reisinger K, Asbe-Vollkopf A,
Hardt-Weinelt K, Kaufmann R. J Invest Dermatol. 2000 July;
115(1):30-6; Kosmadaki M G, Yaar M, Arble B L, Gilchrest B A. FASEB
J. 2003 March; 17(3):466-8; Longuet-Perret I, Schmitt D, Viac J. Br
J Dermatol. 1998 February; 138(2):221-4). VEGF affects a host of
parameters of skin microvasculature; the most prominent being the
increase in permeability of microcapillaries (Dvorak H F, Brown L
F, Dvorak A M. Am J Pathol. 1995 May; 146(5):1029-39). It has been
postulated that VEGF (first known as the Vascular Permeability
Factor) induces microcapillary hyperpermeability through the
loosening of endothelial cell-cell interaction creating
microbreaches through which leukocytes (neutrophils) and plasma
exudate (Harhaj N S, Antonetti D A. Int J Biochem Cell Biol. 2004
July; 36(7):1206-1237).
[0025] Additionally, VEGF secretion by the fibroblasts has been
shown not only to be upregulated by UV, but also by PGE.sub.2
itself (Trompezinski S, Pernet I, Schmitt D, Viae J. Inflamm Res.
2001; (50):422-427). The autocrine/paracrine effect of PGE.sub.2 on
VEGF secretion by skin cells exemplifies the complex cell-cell
communication which exists under stressful conditions. With the
aging process, positive regulation of PGE.sub.2 on VEGF secretion
becomes even more strategic as UV-induced PGE.sub.2 production in
the skin increases as one ages (Seo J Y, Kim E K, Lee S H, Park K
C, Kim K H, Eun H C, Chung J H. Mechanisms of Ageing and
Development. 2003; (124): 903-910). This observation further
illustrates the cross-talks that occur between the biochemical
pathways involved in both chronological aging and actinic aging. A
synergistic superimposition of these two aging modes would
accelerate the loss of integrity of skin microcapillaries (dilation
and hyperpermeability) and exacerbate leukocyte efflux.
[0026] Neutrophils (a subset of leukocytes also referred to as
polymorphonuclear cells) efflux towards the ECM as a result of
microcapillary dilation and hyperpermeability. Evolving scientific
knowledge provides increasing support for the importance of
dermo-epidermal infiltrating neutrophils as effectors in the
process of photo-damage. Neutrophils represent important cellular
sources of not only elastase but also of other known
ECM-degradation enzymes such as the metalloproteinases (MMP-1,
MMP-8 and MMP-9; Rijken F, Kiekens R C M, Bruijnzeel P L B. British
Journal Dermatology. 2005 February; 152(2):321-8). Most of the
emphasis within the scientific community focused around MMPs and
their action in causing the breakdown of collagen fibers and other
ECM macromolecules. However, the action of a specific catabolic
enzyme, elastase, may also impose significant consequences to the
integrity of the ECM and its components. Elastase targets specific
molecular substrates (elastin fibers) that may differ from those
attacked by MMP (mainly collagen fibers), however, the outcome is
similar and translates into the comparable disorganisation of the
skin ECM.
[0027] Human leukocyte elastase (HLE), a broad spectrum serine
protease of 30 kDa, is a specific elastolytic enzyme that is
involved in the turnover of elastic fibers and the remodelling of
the ECM. Elastin fibers are mostly responsible for the resiliency
of the skin's ECM. Even though elastin fibers represent less than
2% of the total dry weight of the skin (in comparison, collagen
fibers comprise more than 70% of total skin dry weight), they
intermingle in functional interactions with other fibrillar
macromolecules and provide the visco-elastic properties required
for normal skin functions.
[0028] Released from a signal source, cytokines initiate the efflux
of neutrophils toward the ECM, seemingly mimicking an inflammatory
reaction. Neutrophils gain access to the signal source by migrating
through the connective tissue thereby destroying encountered
fibrillar components. Once in the connective tissue, neutrophils
cells actively continue the secretion of degradation enzymes
thereby continuing their mission by breaking down elastic fibers of
the ECM. Excessive proteolytic activity of proteolytic enzymes such
as MMPs and neutrophil elastase is known to be associated with
structural alteration of the tridimentional organization of the
ECM. Ultimately, these histological modification deleterious
changes will translate into macroscopic symptoms degeneration
deterioration and become visible in the form of fine lines and
wrinkles; hallmarks of skin aging.
[0029] The importance of elastic fibers for the maintenance of skin
resiliency and elasticity is well exemplified in the many skin
disorders in which the integrity of elastin network is affected
(Lewis K G, Bercovitch L, Dill S W, Robinson-Bostom L. J Am Acad
Dermatol. 2004 July; 51(1):1-21; Lewis K G, Bercovitch L, Dill S W,
Robinson-Bostom L. J Am Acad Dermatol. 2004 August;
51(2):165-85).
[0030] The integrity of the ECM and its dynamic interactions with
skin cells is of primary importance in tissue repair (Midwood K S,
Williams L V, Schwarzbauer J E. Int J Biochem Cell Biol. 2004 June;
36(6):1031-7).
[0031] A relationship has been established between UV exposure,
up-regulation of VEGF, exudation of elastase-producing neutrophils
in the skin, disorganisation of the elastin compartment of the ECM
and the appearance photo-damage (Yano K, Kadoya K, Kajiya K, Hong Y
K, Detmar M. Br J Dermatol. 2005 January; 152(1):115-21). The same
research group (Yano K, Kajiya K, Detmar M. A novel mechanism of
cutaneous photo-damage mediated by angiogenesis and inhibitory
effects of chlorella extract on UV-induced angiogenesis. 23rd IFSCC
Congress. 2004; 46-51) has shown that a Chlorella extract (a green
algae extract) increases trombospondin-1 (TSP-1) expression in UVB
irradiated keratinocytes and prevents UVB-induced predominant
expression of VEGF against TSP-1 in vitro.
[0032] Cellular and molecular mechanisms triggered by UV and
leading to the appearance of clinical signs of actinic aging may
reflect a confused inflammatory and repair elicited response of the
skin in reaction to environmental aggressions.
[0033] Thus, some pathologies are accompanied by a chronic
inflammatory state of the connective tissue in which the balance
between the destruction, repair and resolution phases is upset
which leads to defective reconstruction of the lesioned tissue.
[0034] With this aim, the inventors have studied the action of
various polysaccharide compositions. It is known that specific
polysaccharide compositions, such as glycosaminoglycans,
participate in the composition of the proteoglycans present at the
cell/extracellular matrix interface, and play a role in regulating
cell functions. It is also known that glycosaminoglycans in a
soluble form, for example heparin or dextran derivatives, can
modify cell functions via their interaction with various components
of the ECM.
[0035] Ferrao and Mason (Biochem. Biophys. Acta. 1180, 225-230,
(1993)) have studied the action of various polysaccharides on human
dermal fibroblast proliferation, and indicated that at
concentrations of about 100 .mu.g/ml, heparin, heparan sulfate,
pentosan polysulfate and a fucoidan inhibit this proliferation,
whereas chondroitin sulfate, dermatan sulfate and hyaluronate have
no effect. It is indicated that the inhibitory effect on
proliferation leads to a stimulation of type I collagen synthesis.
Conversely, an inhibition of collagen I synthesis is observed when
the polysaccharides are added to cultures which have reached
confluence.
[0036] Berteau and Mulloy (Glycobiology 13(6): 29R-40R, 2003) have
made a review on fucans, wherein it is said that, like heparin,
they have anti-proliferative effects on vascular smooth muscle
cells and on fibroblasts, in addition to an anti-coagulant effect.
Nothing is disclosed on the activity of fucans on VEGF or on
inflammation mediators, except for TNF-alpha.
[0037] Matsumoto et al. (Clin. Exp. Immunol 136(3): 432-439, 2004)
showed that oral ingestion of fucans from Cladosiphon okamuranus
Tokida (0.05% w/w with food) inhibits the release of Interferon
gamma and IL-6 by colonic lamina propria cells. They propose fucans
as dietary supplement for treating patients with inflammatory bowel
disease.
[0038] Zhang et al. (Zhang O, Li N. Qi H Xu Z. Li Z Phytother Res.
2005 January; 19(1):50-3) reported that elevated urinary protein
excretion and plasma creatinine due to the induction of Heymann
nephritis were significantly reduced by fucoidan oral
administration at doses of 100 and 200 mg/kg, daily. The
renoprotective effect of fucoidan on active Heymann nephritis is a
good indication of its bioavailability after oral
administration.
[0039] Li et al (Li N., Zhang O, Song J. Food Chem Toxicol. 2005
March; 43(3):421-6) investigate the acute and subchronic (6 months)
toxicity of fucoidan extracted from Laminaria japonica in Wistar
rats. Fucoidans did not show significant toxicological changes when
300 mg/kg body weight per day of fucoidan was orally administered.
However, the clotting time was significantly prolonged when the
dose was increased to 900 and 2500 mg/kg body weight per day.
Besides this, no other signs of toxicity were observed. Based on
these results, it can be concluded that no adverse effect level of
fucoidan from L. japonica is observed at or below 300 mg/kg body
weight per day.
[0040] Granert et al. (J. Clin. Invest. 93: 929-936, 1994) disclose
that fucans, administered i.v. (10 mg/Kg body weight) reduce the
accumulation of leukocytes and plasma proteins in the CSF of
rabbits intrathecally challenged with pneumococcal antigen. They
also show that fucans inhibit leukocyte recruitment into an
inflamed tissue site (rabbit skin) thus suggesting that fucans may
be effective when administered in situ or at a distance from the
inflamed site.
[0041] Preobrazhenskaya et al. (Biochem. Mol. Biol. Int. 43(2):
443-451, 1997) show that neutrophil recruitment into an
inflammatory site (rat peritoneum) is reduced by fucans
administered i.v. (0.8 mg). The anti-extravasation effect is quite
remarkable but has a short duration.
[0042] One of the drawbacks of the use of fucans and galactans is
their cytoxicity. Stevan et al (J. Submicrosc. Cytol. Pathol.
33(7): 477-484, 2001) show that fucans, particularly, at a
sulfate/sugar ratio of 1.9 and concentration of 2.5 microgram/mL
cause toxicity in HeLa cells, as seen from the atypical nuclei,
altered cell morphology and impaired cell division.
[0043] There is therefore a need to improve fucans compositions to
increase their efficacy and decrease their toxicity.
[0044] The present invention seeks to meet these needs and other
needs.
[0045] The present description refers to a number of documents, the
content of which is herein incorporated by reference in their
entirety.
SUMMARY OF THE INVENTION
[0046] More specifically, in accordance with the present invention,
there is provided a use of an anti-inflammatory polysaccharides
composition comprising fucans and galactans to inhibit the release
of one or more of IL-8, PGE2 and VEGF by a cell activated during an
inflammatory process.
[0047] In accordance with another aspect of the present invention,
there is provided a use of an anti-inflammatory polysaccharides
composition comprising fucans and galactans in the manufacture of a
medicament to inhibit the release of one or more of IL-8, PGE2 and
VEGF by a cell activated during an inflammatory process.
[0048] In accordance with specific embodiments of these uses, the
release of IL-8, PGE2 and VEGF is from epithelial cells. According
to other specific embodiments, the fucans increase the inhibition
of inflammation, and reduce the cytotoxicity of the galactans.
According to still other specific embodiments, the galactans are
obtained from red algae. According to still other specific
embodiments, the red algae are Asparagopsis armata. According to
still other specific embodiments, the fucans are obtained from
brown algae. According to still other specific embodiments, the
brown algae are Ascophyllum nodosum. According to still other
specific embodiments, the galactans have an average molecular
weight higher than about 100 kDa. According to still other specific
embodiments, the galactans have an average molecular weight of
about 350 kDa. According to still other specific embodiments, the
galactans have a galactose content of about 37% of dry weight of
the galactans. According to still other specific embodiments, the
galactans have an uronic acid content of about 3% of dry weight of
the galactans. According to still other specific embodiments, the
galactans have a sulfate content of about 27% of dry weight of the
galactans. According to still other specific embodiments, the
fucans have an average molecular weight ranging from about 0.1 kDa
to about 100 kDa. According to still other specific embodiments,
the fucans have an average molecular weight ranging from about 5
kDa to about 25 kDa. According to still other specific embodiments,
the fucans have a fucose content of about 20-35% of dry weight of
the fucans. According to still other specific embodiments, the
fucans have an uronic acid content of about 10 to about 29% of dry
weight of the fucans. According to still other specific
embodiments, the fucans have a sulfate content of about 15 to about
25% of dry weight of the fucans. According to still other specific
embodiments, the ratio fucans/galactans present in the composition
is between about 2.5/1 (w/w) to about 40/1 (w/w). According to
still other specific embodiments, the fucans and the galactans are
present a ratio of about 10 fucans/1 galactans (w/w) in the
composition. According to still other specific embodiments, the
galactans comprise native galactans. According to still other
specific embodiments, the fucans comprise native fucans. According
to still other specific embodiments, the fucans comprise
depolymerized fucans. According to still other specific
embodiments, the fucans comprise demineralized fucans. According to
still other specific embodiments, the galactans comprise
demineralized galactans. According to still other specific
embodiments, the use is a topical use. According to still other
specific embodiments, the use is to alleviate or improve skin
disorders or conditions caused by UV exposure, chemical stress or
aggression from pollutants, exfoliating agents or skin irritants.
According to still other specific embodiments, the use is to
prevent skin disorders or conditions caused by UV exposure,
chemical stress or aggression from pollutants, exfoliating agents
or skin irritants.
[0049] In accordance with another aspect of the present invention,
there is provided an anti-inflammatory composition comprising a
ratio of brown algae fucans/red algae galactans of between about
2.5/1 (w/w) to about 40/1 (w/w), the galactans having a molecular
weight higher than about 100 kDa, and the fucans having a molecular
weight between about 0.1 kDa and 100 kDa. According to specific
embodiments, the galactans have an average molecular weight of
about 350 kDa. According to other specific embodiments, the
galactans have a galactose content of about 37% of dry weight of
the galactans. According to still other specific embodiments, the
galactans have an uronic acid content of about 3% of dry weight of
the galactans. According to still other specific embodiments, the
galactans have a sulfate content of about 27% of dry weight of the
galactans. According to still other specific embodiments, the
fucans have an average molecular weight ranging from about 5 kDa to
about 25 kDa. According to still other specific embodiments, the
fucans have a fucose content of about 20-35% of dry weight of the
fucans. According to still other specific embodiments, the fucans
have an uronic acid content of about 10 to about 29% of dry weight
of the fucans. According to still other specific embodiments, the
fucans have a sulfate content of about 15 to about 25% of dry
weight of the fucans. According to still other specific
embodiments, the ratio of brown algae fucans/red algae galactans
being of 10/1. According to still other specific embodiments, the
galactans having a molecular weight of about 350 kDa, and the
fucans having a molecular weight between about 15 kDa and 25 kDa.
According to still other specific embodiments, the brown algae is
Ascophyllum nodosum. According to still other specific embodiments,
the red algae is Asparagopsis armata. According to still other
specific embodiments, the galactans comprise native galactans.
According to still other specific embodiments, the fucans comprise
native fucans. According to still other specific embodiments, the
fucans comprise depolymerized fucans. According to still other
specific embodiments, the fucans comprise demineralized fucans.
According to still other specific embodiments, the galactans
comprise demineralized galactans. According to still other specific
embodiments, the composition is a topical composition. According to
still other specific embodiments, the composition is a cosmetic
composition.
[0050] In accordance with another aspect of the present invention,
there is provided a method of inhibiting the release of one or more
of IL-8, PGE2 and VEGF by a cell activated during an inflammatory
process comprising administering to a subject a polysaccharides
composition comprising fucans and galactans. According to specific
embodiments of the method, the polysaccharides composition is
topically administered to the subject. According to other specific
embodiments of the method, the subject is a human affected by skin
disorders or conditions caused by UV exposure, chemical stress or
aggression from pollutants, exfoliating agents or skin
irritants.
[0051] As used herein the term "fucans" refers to polysaccharides
comprising sulfated fucose and uronic acid, wherein the
polysaccharides have antiinflammatory and anti-extravasation
activities. Without being so limited, fucans derived from
Laminariales, Chordariales, Fucales marine algae including
Kjellmaniella crassifolia, Laminaria japonica, Kjellmaniella,
Fucus, Nemacystus, Cladosiphon okamuranus, Undaria, Undaria
pinnatifida (Wakame Mekabu), Ecklonia kurome, Eisenia, Ecklonia,
Giant kelp, Lessonia nigrescence Gelidiun amansii, Gracilaria,
Pteroclavia capillacae and Ascophyllum nodosum along with those
derived from Echinodermata, sea cucumber, Echnoidea and Asterozoa
are suitable for uses and methods of the present invention. Fucans
of the present invention are exemplified herein by fucans derived
from the brown algae Ascophyllum nodosum.
[0052] As used herein the term "galactans" refers to
polysaccharides comprising sulfated beta-D-galactose, xylose and
uronic acid. Without being so limited, Rhodomelaceae,
Corallinaceae, Gracilariales, Ceramiales, Gigartinales marine
algae, including Asparagopsis armata, Bostrychia maontanei,
Corallina species, Polysiphonia lanosa, Gracilaria species,
Acanthophora spicifer, Georgiella confluens, Laurencia coronopus,
Porphyra haitanensis, Botryocladia occidentalis, Chondrus
ascellatus, Gymnogongrus tolurus, Phacelocarpus peperocarpos,
Sargassum kjellmanianum, contain galactans suitable for uses and
methods of the present invention. The galactans of the present
invention are exemplified herein by galactans derived from the red
algae Asparagopsis armata.
[0053] The term "about" when used in relation to ranges apply to
both ends of the range. It is used to reflect the relative
precision of the equipment and process used to obtain and to
characterize the compositions of the present invention.
[0054] The articles "a," "an" and "the" are used herein to refer to
one or to more than one (i.e., to at least one) of the grammatical
object of the article.
[0055] The term "including" and "comprising" are used herein to
mean, and re used interchangeably with, the phrases "including but
not limited to" and "comprising but not limited to".
[0056] The term "such as" is used herein to mean, and is used
interchangeably with, the phrase "such as but not limited to".
[0057] As used herein the term "subject" is meant to refer to any
mammal including human, mice, rat, dog, cat, pig, cow, monkey,
horse, etc. In a particular embodiment, it refers to a human.
[0058] As used herein the term "anti-inflammatory" with regards to
the polysaccharides compositions of the present invention relate to
their ability to inhibit VEGF and/or IL-8 and/or PGE2.
[0059] Applications of the polysaccharides compositions of the
present invention include topically applicable cosmetic
compositions. Non-limitative examples of such topically applicable
compositions include skin care cream, cleansing cream, skin care
lotion, skin care gel, skin care foam, sun care composition,
make-up removal cream, make-up removal lotion, foundation cream,
liquid foundation, bath and shower preparation, deodorant
composition, antiperspirant composition, shaving products
composition, after-shave gel or lotion, beauty aids composition,
depilatory cream, soap composition, hand cleaner composition,
cleansing bar, baby care, hair care, shampoo, setting lotion,
treatment lotion, hair cream, hair gel, coloring composition,
restructuring composition, permanent composition, anti-hair loss
composition, or any other composition which is adapted for the use
in a topical cosmetic regimen.
[0060] Polysaccharides compositions of the present invention may
comprise at least one additional active ingredient. Without being
so limited, such additional active ingredient may modulate at least
one of cell differentiation, cell metabolic activity, cell
structure, cell proliferation, extracellular processes and
pigmentation. Without being so limited, the polysaccharides
compositions of the present invention may further comprise at least
one of an anesthesic agent, anti-acne agent, anti-aging agent,
antibacterial agent, anticellulite agent, antifungal agent,
anti-inflammatory agent, anti-irritant agent, antioxidant agent,
antiparasitic agent, antipollution agent, antipruritic agent,
anti-rosacea agent, anti-seborrhea agent, anti-stress agent,
anti-telangiectasia agent, antiviral agent, anti-wrinkle agent,
baby care agent, bath and body agent, calming agent, cleansing
agent, collagen synthesis agent, elastase inhibitory agent,
exfoliant agent, facial peeling agent, firming agent, foot care
agent, free radical scavenging agent, immune function modulator
agent, keratolytic agent, lift agent, make-up remover agent,
melanogenesis stimulator agent, matrix metalloproteinase inhibitory
agent, moisturizing agent, oil absorbent agent, osmoregulator
agent, anti-photoaging agent, protecting agent, rejuvenating agent,
regenerating agent, restructuring agent, sensitive skin agent,
shaving product agent, skin defense enhancer agent, skin clarifier
agent, skin repair agent, slimming agent, smoothing agent,
softening agent, soothing agent, sun care agent, sunless tanning
agent, tensing agents and whitening agent, or any other agent
adapted for use in a cosmetic regimen that comprises topical
application of said cosmetic composition, and which complements or
supplements the effect of the polysaccharides of the present
invention.
[0061] Without being so limited, agents that modulate cell
differentiation or proliferation include plant extracts, algae
extracts, fruit extracts, vegetable extracts, leguminous plant
extracts, ferments, proteolytic hydrolysates, peptides, yeast
extracts and its derivatives, microorganism extracts, animal
derivative extracts and synthetic compounds. More particularly,
such agents include retinoic acid and its derivatives (retinol,
retinaldehyde, retinyl palmitate, trans-retinoic acid, 13-cis
retinoic acid, 9-cis retinoic acid, retinoyl glucuronoides,
tretinoin, isotretinoin, etretinate, acitretine, tazarotene,
adapalene, .beta.-carotene, retinyl ester), vitamin D and its
derivatives (cholecalciferol, ergocalciferol,
25-hydroxycholecalciferol), growth factors and estradiol
derivatives.
[0062] Without being so limited, anaesthesics include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include lidocaine chlorhydrate and its
derivatives.
[0063] Without being so limited anti-acne agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include benzoyl peroxide, retinoic acid
and its derivatives (retinol, retinaldehyde, retinyl palmitate,
trans-retinoic acid, 13-cis retinoic acid, 9-cis retinoic acid,
retinoyl glucuronoides, tretinoin, isotretinoin, etretinate,
acitretine, tazarotene, adapalene, .beta.-carotene, retinyl ester),
salicylic acid, sulfur, sulfurated lime, alcohol and acetone.
[0064] Without being so limited, anti-aging/anti-wrinkle agents
include plant extracts, algae extracts, fruit extracts, vegetable
extracts, leguminous plant extracts, ferments, proteolytic
hydrolysates, peptides, yeast extracts and its derivatives,
microorganism extracts, animal derivative extracts and synthetic
compounds. More particularly, such agents include hyaluronic acid,
sodium-2-pyrrolidone carboxylate, glycosaminoglycans, kinetin,
retinoic acid and its derivatives (retinol, retinaldehyde, retinyl
palmitate, trans-retinoic acid, 13-cis retinoic acid, 9-cis
retinoic acid, retinoyl glucuronoides, tretinoin, isotretinoin,
etretinate, acitretine, tazarotene, adapalene, .beta.-carotene,
retinyl ester), epidermal growth factor, ceramide,
ethylbisiminomethylguaiacol manganese chloride, glycation
inhibitors, chrysanthellum indicum extract and aphanizomenon flos
aquae extract.
[0065] Without being so limited, antibacterial agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include eucalyptus extract, clindamycin
phosphate, cavacrol, erythromycin and antibiotics belonging to the
group of tetracyclines.
[0066] Without being so limited, antifungal agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include econazole, ketoconazole,
miconazole, amphotericin B, terbinafine and octopirox.
[0067] Without being so limited, anti-inflammatory agents include
plant extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include allantoin, vitamin E and its
derivatives (.alpha.-tocopherol, .delta.-tocopherol,
.gamma.-tocopherol), chamomile oil, gingko biloba oil and camellia
sinensis extract.
[0068] Without being so limited,
anti-irritant/soothing/smoothing/calming agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include allantoin, camellia sinensis
extract, lavender oil, aloe vera, linden extract, epilobium
angustifolium extract, chysanthellum indicum extract, cola nitida
extract and alteromonas ferment extract.
[0069] Without being so limited, antioxidant agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include furfuryladenine, panthenol,
lipoic acid, ubiquinone, niacinamide, melatonin, catalase,
glutathione, superoxide dismutase, polyphenols, cysteine,
allantoin, kinetin, vitamin C and its derivatives (ascorbyl
palmitate, magnesuim ascorbyl phosphate, sodium ascorbyl
phosphate), vitamin E and its derivatives (.alpha.-tocopherol,
.delta.-tocopherol, .gamma.-tocopherol), grape seed extract and
camellia sinensis extract.
[0070] Without being so limited, antipruritic agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include thenaldine, trimeprazine,
cyproheptadine.
[0071] Without being so limited, anti-rosacea/anti-telangiectasia
agents include plant extracts, algae extracts, fruit extracts,
vegetable extracts, leguminous plant extracts, ferments,
proteolytic hydrolysates, peptides, yeast extracts and its
derivatives, microorganism extracts, animal derivative extracts and
synthetic compounds. More particularly, such agents include
metronidazole, vasoconstrictors, benzoyl peroxide, azelaic acid,
sulphur, soy proteins and glycosaminoglycans.
[0072] Without being so limited, anti-seborrhea agents include
plant extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include progesterone derivatives,
isoleutrol and hinokitiol.
[0073] Without being so limited, sensitive skin agents include
plant extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include rose oil and jasmine oil.
[0074] Without being so limited, cleansing agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include ammonium lauryl sulfate, ammonium
laureth sulfate, cocamide MEA, triethanolamine lauryl sulfate,
sodium stearate and nettle leaf extract.
[0075] Without being so limited, collagen synthesis agents include
plant extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include retinoic acid and its derivatives
(retinol, retinaldehyde, retinyl palmitate, trans-retinoic acid,
13-cis retinoic acid, 9-cis retinoic acid, retinoyl glucuronoides,
tretinoin, isotretinoin, etretinate, acitretine, tazarotene,
adapalene, .beta.-carotene, retinyl ester), vitamin C and its
derivatives (ascorbyl palmitate, magnesium ascorbyl phosphate,
sodium ascorbyl phosphate), growth factors and its derivatives.
[0076] Without being so limited, exfoliant agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include alpha/beta hydroxy acids,
salicylic acid, glycolic acid, lactic acid, citrus acid and walnut
shell powder.
[0077] Without being so limited, facial peeling agents include
plant extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include glycolic acid, lactic acid,
trichloroacetic acid and phenol.
[0078] Without being so limited, firming/tensing agents include
plant extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include dimethylaminoethanol,
neuro-cosmetic actives (Botox.TM.-like), chitosan, arnica extract,
fennel-sweet oil and papaya extract.
[0079] Without being so limited, free radical
scavenging/antipollution/anti-stress agents include plant extracts,
algae extracts, fruit extracts, vegetable extracts, leguminous
plant extracts, ferments, proteolytic hydrolysates, peptides, yeast
extracts and its derivatives, microorganism extracts, animal
derivative extracts and synthetic compounds. More particularly,
such agents include grape seed extract, alpha-tocopherol and the
esters thereof, superoxide dismutase, some chelating agents of
metals, vitamin C and its derivatives (ascorbyl palmitate,
magnesium ascorbyl phosphate, sodium ascorbyl phosphate).
[0080] Without being so limited, hair care agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include poly-D-glucosamine,
poly-N-acetyl-D-glucosamine, stearalkonium chloride and
triethanolamine lauryl sulfate.
[0081] Without being so limited, matrix metalloproteinase
inhibitory agents include plant extracts, algae extracts, fruit
extracts, vegetable extracts, leguminous plant extracts, ferments,
proteolytic hydrolysates, peptides, yeast extracts and its
derivatives, microorganism extracts, animal derivative extracts and
synthetic compounds. More particularly, such agents include
camellia sinensis extract, polyphenols, spatholobi caulis extract,
euonymus alatus extract, rhizoma notopterygii extract, quercetin,
glycosaminoglycans, polymethoxy flavonoid, N-acetyl-cysteine,
2-furildioxime, isoflavone, vitamin C and its derivatives (ascorbyl
palmitate, magnesium ascorbyl phosphate, sodium ascorbyl
phosphate), retinoic acid and its derivatives (retinol,
retinaldehyde, retinyl palmitate, trans-retinoic acid, 13-cis
retinoic acid, 9-cis retinoic acid, retinoyl glucuronoides,
tretinoin, isotretinoin, etretinate, acitretine, tazarotene,
adapalene, .beta.-carotene, retinyl ester) and hydroxamate
derivatives.
[0082] Without being so limited, moisturizing agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include cucumber extract,
sodium-2-pyrrolidone carboxylate, sodium PCA, sodium hyaluronate,
chitin and its derivatives, alpha hydroxy acids, hyaluronic acid
and hydrolysed wheat protein.
[0083] Without being so limited, osmoregulator agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include mannitol, dulcitol and
betaine.
[0084] Without being so limited, protecting agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include poly-N-acetyl-D-glucosamine,
poly-D-glucosamine, alkyloamides, chitosan, chrysanthellum indicum
extract, camellia sinensis extract and alteromonas ferment
extract.
[0085] Without being so limited, rejuvenating agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include rosemary extract, rosewood
extract, geranium extract and vitamin E and its derivatives
(.alpha.-tocopherol, .delta.-tocopherol, .gamma.-tocopherol).
[0086] Without being so limited, skin repair agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include retinoic acid and its derivatives
(retinol, retinaldehyde, retinyl palmitate, trans-retinoic acid,
13-cis retinoic acid, 9-cis retinoic acid, retinoyl glucuronoides,
tretinoin, isotretinoin, etretinate, acitretine, tazarotene,
adapalene, .beta.-carotene, retinyl ester), allantoin, eucalyptus
extract, lavender oil, rose oil and activators of collagen
synthesis and activators of components of the skin's extracellular
matrix.
[0087] Without being so limited, slimming/anticellulite agents
include plant extracts, algae extracts, fruit extracts, vegetable
extracts, leguminous plant extracts, ferments, proteolytic
hydrolysates, peptides, yeast extracts and its derivatives,
microorganism extracts, animal derivative extracts and synthetic
compounds. More particularly, such agents include chrysanthellum
indicum extract, dihydromyricetin, theobromine, theophylline,
aminophylline, caffeine, isopropylarterenol hydrochloride,
epinephrine, .alpha.-MSH agonists, adenylate cyclase activators and
phosphodiesterase inhibitors.
[0088] Without being so limited, sun care/photo aging agents
include plant extracts, algae extracts, fruit extracts, vegetable
extracts, leguminous plant extracts, ferments, proteolytic
hydrolysates, peptides, yeast extracts and its derivatives,
microorganism extracts, animal derivative extracts and synthetic
compounds. More particularly, such agents include PABA
(p-aminobenzoic acid) and derivatives, gluconolactone, salicylates,
cinnamates, benzophenones, dibenzoylmethanes, oxybenzone, vitamin E
and its derivatives (.alpha.-tocopherol, .delta.-tocopherol,
.gamma.-tocopherol), ethylbisiminomethylguaiacol manganese
chloride, glycosaminoglycans, retinoic acid and its derivatives
(retinol, retinaldehyde, retinyl palmitate, trans-retinoic acid,
13-cis retinoic acid, 9-cis retinoic acid, retinoyl glucuronoides,
tretinoin, isotretinoin, etretinate, acitretine, tazarotene,
adapalene, .beta.-carotene, retinyl ester), titanium dioxide, octyl
methoxycinnamate, benzophenone, octyl salicylate, epilobium
angustifolium extract, rumex occidentalis extract, chrysanthellum
indicum extract, camellia sinensis extract and alteromonas ferment
extract.
[0089] Without being so limited, sunless tanning/melanogenesis
stimulator agents include plant extracts, algae extracts, fruit
extracts, vegetable extracts, leguminous plant extracts, ferments,
proteolytic hydrolysates, peptides, yeast extracts and its
derivatives, microorganism extracts, animal derivative extracts and
synthetic compounds. More particularly, such agents include
dihydroxyacetone, .alpha.-MSH agonists, adenylate cyclase
activators and phosphodiesterase inhibitors.
[0090] Without being so limited, toning agents include plant
extracts, algae extracts, fruit extracts, vegetable extracts,
leguminous plant extracts, ferments, proteolytic hydrolysates,
peptides, yeast extracts and its derivatives, microorganism
extracts, animal derivative extracts and synthetic compounds. More
particularly, such agents include nettle extract, orange blossom
extract, rosewood extract and witch hazel extract.
[0091] Without being so limited, whitening/pigmentation agents
include plant extracts, algae extracts, fruit extracts, vegetable
extracts, leguminous plant extracts, ferments, proteolytic
hydrolysates, peptides, yeast extracts and its derivatives,
microorganism extracts, animal derivative extracts and synthetic
compounds. More particularly, such agents include arbutin, azealeic
acid, vitamin C and its derivatives (ascorbyl palmitate, magnesuim
ascorbyl phosphate, sodium ascorbyl phosphate), hydroquinone,
N-acetyl-4-S-cysteanimylphenol, kojic acid, melanostat
(melanostatine), tretinoin, retinoic acid and its derivatives
(retinol, retinaldehyde, retinyl palmitate, trans-retinoic acid,
13-cis retinoic acid, 9-cis retinoic acid, retinoyl glucuronoides,
tretinoin, isotretinoin, etretinate, acitretine, tazarotene,
adapalene, .beta.-carotene, retinyl ester), ruminex occidentalis
extract, licorice, mulberry, arctostaphylos uva-ursi (bearberry),
tyrosinase inhibitors, melanosome-transfer inhibitors and melanin
scavengers.
[0092] The polysaccharides composition of the present invention may
be formulated so as to provide for a specifically controlled
delivery system. Non-limitative examples of such delivery systems
include slow delivery system, rapid delivery system, immediate
delivery system, delayed delivery system, zero-order delivery
system and dual or multiple speed delivery system. Such controlled
delivery systems may be achieved with specific formulations
including chemical delivery systems, multiple emulsions,
microemulsions, nanoemulsions, encapsulations such as liposomes,
microspheres, nanospheres, microsponges, beads and cyclodextrins,
polymeric matrices, polymeric cosmetic conjugates, oil
body/oleosin, oil-soluble molecular film, skin patches, unit
dosages.
[0093] The polysaccharides compositions of the present invention
may be formulated into a cosmetically acceptable vehicle including
a solution, dispersion, lotion, serum, microgranulate dispersion,
vesicular ionic or non-ionic dispersion, alcoholic or
hydro-alcoholic aqueous solution, cream, gel, oil-in-water or
water-in-oil emulsion, foam, aerosol, solid or paste.
[0094] The polysaccharides compositions of the present invention
may further comprise additional excipients such as buffer agent,
carrier agent, chelating agent, conditioner agent, coloring agent,
detackifier agent, emollient agent, emulsifier agent, film former
agent, foaming agent, humectant agent, lactylate agent, lipophilic
agent, lubricant agent, neutralizer agent, oil agent, opacifier
agent, preservative agent, solubilizer agent, solvent agent,
stabilizer agent, surfactant agent, thickener agent, viscosity
agent, water absorbent agent and wetting agent.
[0095] Without being so limited, buffer agents are salts of
bases/acids, compatible with the nature of the skin and with its
pH. Sodium acetate is an example of a frequently used buffer
agent.
[0096] Without being so limited, carrier agents are ingredients
capable of aiding the application of the active ingredient.
Isohexadecane is an example of a frequently used carrier.
[0097] Without being so limited, chelating agents are ingredients
capable of binding mono and divalent cations, such as tetrasodium
EDTA and disodium EDTA.
[0098] Without being so limited, conditioner agents are ingredients
with lubricating action and hydrating effect, such as cetrimonium
chloride, dicetyldimonium chloride, trideceth-I2, quaternium-Z7,
quaternium-I8, polyquaternium-10, behentrimonium methosulfate,
cetearyl alcohol, stearamidopropyl dimethylamine,
trimethylsilylamodimethicone, isolaureth-6, octoxynol-4,
dimethicone, dimethiconol, cyclopentasiloxane, pareth-7, pareth-9,
linoleic acid and glycerin.
[0099] Without being so limited, detackifier agents are ingredients
capable of adsorbing onto tacky materials and reduce their tendency
to adhere, such as cyclopentasiloxane, dimethicone and vinyl
dimethicone, phenyl trimethicone, isopropyl esters, isostearate
esters, dimethyl sebacate and dipropyl sebacate.
[0100] Without being so limited, emollient agents are ingredients
with lubricating action and hydrating effect, such as isopropyl
palmitate, sunflower seed oil, mineral oil, stearyl stearate,
isopropyl myristate, lanolin, caprylic, capric triglyceride,
cyclopentasiloxane, dimethicone, vinyl dimethicone,
bis-phenylpropyl dimethicone, alkyl dimethicone, sorbitan stearate,
sucrose distearate, myristyl alcohol, myristyl lactate, cetyl
acetate, dicaprylyl ether, floraester-20, maleated soybean oil,
cyclomethicone, shea butter, hydrogenated coconut oil, isopropyl
palmitate, diisostearoyl trimethylolpropane siloxy silicate and
alkyl benzoate.
[0101] Without being so limited, emulsifier agents are ingredients
capable of preventing the separation of immiscible substances in an
emulsion, of helping to distribute evenly one substance in another,
of improving texture, homogeneity, consistency and stability, such
as cetearyl alcohol, glyceryl stearate, alkyl acrylate
crosspolymer, stearic acid, emulsifying wax, sorbitan oleate,
sorbitan stearate, polysorbate, polyethylene glycopolysorbate,
triethanolamine, cyclopentasiloxane, dimethicone copolyol, PEG-30
dipolyhydroxystearate, sucrose distearate, PEG-100 stearate, sodium
dioctylsulfosuccinate, polyacrylamide, isoparaffin, laureth-7,
cetyl phosphate, DEA cetyl phosphate, glycol stearate, stearyl
alcohol, cetyl alcohol, behentrimonium methosulfate and
ceteareth-2.
[0102] Without being so limited, film former agents are ingredients
capable of forming a dimensionally stable and continuous film to
minimize the formula tackiness, such as wheat protein, eicosene
copolymer, perfluoromethylisopropyl ether, diisostearoyl
trimethylol propane siloxy silicate, trimethylsiloxysilicate,
dimethicone, vinyl dimethicone and cyclopentasiloxane.
[0103] Without being so limited, foaming agents are ingredients
capable of regulating the amount of air in a product, such as
lauramide DEA and cocamide MEA, disodium laureth sulfosuccinate,
disodium N-octadecyl sulfosuccinamate, ammonium lauryl sulphate,
triethanolamine lauryl sulfate, sodium lauryl sulphate and sodium
2-ethyl hexylsulfate.
[0104] Without being so limited, humectant agents are ingredients
capable of maintaining constant humidity and retaining moisture,
such as glycerine, PEG-8, butylene glycol and propylene glycol.
[0105] Without being so limited, lubricant agents are ingredients
capable of adding slipperiness and reducing friction to improve
application, such as dimethicone and dimethicone copolyol.
[0106] Without being so limited, neutralizer agents are ingredients
capable of changing the acid-alkaline balance, such as
triethanolamine and sodium hydroxide.
[0107] Without being so limited, opacifier agents are ingredients
capable of changing the look of a clear or translucent product to a
creamier or pearlier one, such as glyceryl stearate and PEG-100
stearate.
[0108] Without being so limited, preservative agents are
ingredients capable of retarding or preventing microbial or
chemical spoilage and protecting against discoloration, such as
DMDM hydantoin, methylparaben, propylparaben, phenoxyethanol,
ethylparaben, butylparaben, imidazolidinyl urea, diazolidinyl urea,
quaternium-8, quaternium-14, quaternium-15, propylene glycol,
dehydroacetic acid, methylchloroisothiazolinone,
methylisothiazolinone and germaben.
[0109] Without being so limited, solubilizer agents are ingredients
capable of allowing incompatible ingredients to become part of a
homogeneous solution, such as polysorbate, ceteareth, steareth and
PEG.
[0110] Without being so limited, stabilizer agents are ingredients
capable of maintaining physical and chemical properties during and
after processing, preventing or limiting changes in the physical
properties of a substance during product life, such as
polyethylene, sodium chloride, stearyl alcohol, xanthan gum,
tetrasodium EDTA and dimethicone copolyol.
[0111] Without being so limited, surfactant agents are ingredients
capable of reducing surface tension when dissolved in water or a
water solution, reducing interfacial tension between two liquids or
between a liquid and a solid, such as sodium dioctylsulfosuccinate,
octoxynol-40, isolaureth-6, ammonium lauryl sulfate, lauryl
alcohol, lauramide DEA and cocoamidopropyl betaine.
[0112] Without being so limited, thickener agents are ingredients
capable of absorbing water to impart body, improve the consistency
or texture, and stabilize an emulsion, such as stearic acid,
magnesium aluminum silicate, carbomer, alkyl acrylate crosspolymer,
polyacrylamide, isoparaffin, laureth-7, cetyl alcohol, xanthan gum,
alkyl dimethicone, hydroxyethylcellulose, glyceryl stearate,
pentaerythrityl tetrastearate, stearyl alcohol and
polyquaternium-10.
[0113] Without being so limited, viscosity agents are ingredients
capable of controlling the degree of fluidity and the internal
resistance to flow exhibited by a fluid, such as magnesium aluminum
silicate, caprylyl glycol and myristyl alcohol.
[0114] Without being so limited, water absorbent agents are
ingredients capable of absorbing the product's water to maintain
the moisture, such as carboxyvinyl polymer, acrylic copolymer,
polyacrylamide, polysaccharides, natural gum, clay, modified clay,
metallic salt and fatty acid.
[0115] Without being so limited, wetting agents are ingredients
capable of reducing the surface tension of the water for better
penetration or spread over the surface, such as caprylate, caprylyl
glycol, glyceryl caprate, polyglyceryl-2 caprate, polyglyceryl-6,
polyglyceryl-3 laurate and TEA-laureth sulfate.
[0116] Other objects, advantages and features of the present
invention will become more apparent upon reading of the following
non-restrictive description of specific embodiments thereof, given
by way of example only with reference to the accompanying
drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0117] In the appended drawings:
[0118] FIG. 1 depicts the effect of a fucans/galactans composition
of the present invention on VEGF expression in human keratinocytes
in vitro;
[0119] FIG. 2 depicts the effect of a fucans/galactans composition
of the present invention on PGE.sub.2 expression in human
keratinocytes in vitro;
[0120] FIG. 3 depicts the preventive effect of a fucans/galactans
composition of the present invention against skin UV exposure;
and
[0121] FIG. 4 depicts the effect of a fucans/galactans composition
of the present invention after skin UV exposure.
DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
Preparation of Fucans
[0122] Fucans from various algae may be prepared by known methods
and the resulting purified products, fucans-containing products can
be used in the present invention.
[0123] In a specific embodiment, fucans of the present invention
are obtained from Ascophyllum nodosum. In their native form, fucans
are constituted by a heterogenous population of molecules
principally composed of sulfated L-fucose polymers that have high
molecular weights, and are associated with uronic acids. In
addition, several studies clearly show that Ascophyllum nodosum
fucans possess large portion of both .alpha.(1.fwdarw.3) and
.alpha.(1.fwdarw.4) glycosidic bonds.
Native Fucans (i.e. Which Naturally Contain Minerals)
[0124] Fresh, frozen or dried clean algae can be transformed into a
fine powder. Fresh and frozen algae are preferably chopped to
obtain a particle size smaller than about 8 mm, preferably between
about 0.5 mm and about 8 mm, most preferably about between about 2
and about 5 mm.
[0125] The algae could be chopped with instruments including but
not limited to, a meat chopper, kitchen homogenizer, Polytron.TM.
disintegrator and any commercial cutter. Variation and adjustment
of the crushing parameters are well within the knowledge of the
skilled artisan, merely depending on the volume of homogenate and
of the equipment used.
[0126] Fucans may be extracted from the chopped algae by an
adequate volume of water-based solution. The volume of solution
used can be increased without bearing any deleterious effect on the
recovery yield of valuable components. A small volume is preferred
since it is more convenient to manipulate than larger volumes.
Water may be purified by inverse osmosis and multiple filtrations
down to 0.1 micron filter. Many aqueous solutions (containing
salts, for example) could be used in lieu of water. When recovery
of a plurality of hydrosoluble activities is contemplated, working
at a near neutral pH (5.0 to 8.0) and in non-denaturing conditions
is preferred to avoid degradation or denaturation of some of the
active components. For the sake of clarity, any extraction medium
that is compatible with the preservation of biologically active
fucans components is within the scope of this invention. Therefore,
performing the extraction in pure water is preferred. Other
preferred embodiments include those wherein salts and/or chaotropic
agents are added to the water prior to or during extraction.
[0127] The extraction may be performed at about 60 to about 95
degrees Celsius, preferably between about 80 to 95 degrees Celsius.
The extraction time is generally about 12 to 24 hours, preferably
between about 10 and about 16 hours. The speed of the agitation as
well as the volume of aqueous solution may influence both time and
yield of extraction.
[0128] The supernatant is then separated from the pellet. The
separation can be quickly performed by filtration (with a mesh of
about 50 micrometer), centrifugation, or with a commercial
decanter. Variation and adjustment of the separation parameters are
well within the knowledge of the skilled artisan, merely depending
on the volume of homogenate and of the equipment used.
[0129] The resulting supernatant is then clarified to get rid of
fine suspension susceptible of affecting the performance of the
process. The clarification is performed at a temperature ranging
from about 50 to about 80 degrees Celsius, preferably at about 40
to about 60 degrees Celsius
[0130] The clarified material is then cooled down at a temperature
ranging from about 15 to about 30 degree Celsius, preferably from
about 15 to about 25 degrees Celsius.
[0131] The clarified material is then acidified at about pH 2 to
precipitate proteins and alginates. For example, the acidification
could be performed in presence of sulfuric acid or hydrochloric
acid. The acidified materiel is then clarified as described above.
The solution is then neutralized.
[0132] The neutralized solution is then concentrated. This step can
be performed by different means including but not limited to:
dialysis, chromatography (adsorption, ionic exchange, gel
filtration), electrophoresis, ultrafiltration, ultra centrifugation
with zonal density gradients, adsorption and extraction.
[0133] In a specific embodiment, the solution may be ultrafiltered
at about 4 degrees Celsius to 25 degrees Celsius (although
temperature could be increased to about 40 degrees Celsius) on a
tangential flow filtration column. Column membrane porosity is
about 500 kDa, preferably 300 kDa, most preferably 100 kDa. The
obtained fraction is then dialyzed which allows a first
fractionated extract to be obtained, comprising hydrosoluble fucans
of an average molecular weight generally lower than the membrane
cut off taking into account the variability inherent to the
membrane pores and to the conformation of the molecules in the
extraction solvent. This inherent variability applies to any
membrane used herein. This fraction will be further referred to as
the native fucans fraction which is naturally mineralized
(FHMW-M).
[0134] Molecular analysis of fucan fractions reveal that fucose
constitutes between about 15 and 45% and more preferably between 20
and 35% of the dry weight. Moreover, 3 to 35% and more preferably
10 to 29% of the dry weight is composed of uronic acid. These
fucans are richly sulfated, since 12 to 30% and more preferably 15
to 25% of the dry weight is composed of sulfates (SO.sub.4)
radical.
[0135] According to another embodiment, fucans could be extracted
from the chopped algae by an adequate volume of water-based
solution in presence of 1% calcium chloride. The extraction is
performed at about 85 degree C. for about 4 hours. These conditions
allow the proteins comprising alginates to precipitate. The
resulting supernatant is collected, the polysaccharides are
precipitated in the presence of ethanol and the fucans are further
purified by ultrafiltration.
Depolymerized Fucans
[0136] The fucans could be used entirely or partly in their
depolymerized form. In a specific embodiment, the fucans'
depolymerization is performed by acidic hydrolysis. Native fucans
(FHMW-M) are acidified to a pH of about 0.5 to about 2.0,
preferably to about 0.5 to about 1.0. The acidified solution is
incubated at temperature of about 55 to about 70 degree C. for
about 4 hours. Depolymerization may be achieved by different
methods including but not limited to: ultrasounds; UV radiations;
ozonolysis; chemical, radical, or enzymatic hydrolysis; and high
pressure and temperature.
[0137] The solution is then neutralized with NaOh and then
concentrated/purified by ultrafiltration followed by a
dialysis.
[0138] The fucans present in the solution are then precipitated in
the presence of alcohol and the precipitate is dried. Variation and
adjustment of the parameters for alcohol precipitation and drying
are well within the knowledge of the skilled artisan, merely
depending on the volume of the solution and on the equipment used.
The fraction obtained contains hydrosoluble fucans of an average
molecular weight ranging between about 5 kDa and about 25 kDa. This
fraction will be further referred to as the mineralized
depolymerized fucan fraction (FLMW-M).
Demineralised Fucans
[0139] The fucans fractions could further be used entirely or
partly in their demineralised form. Demineralisation could be
industrially performed, such as with but not limited to
nanofiltration. The nanofiltration process is a reverse osmosis
process using a relatively open RO membrane, allowing water and
small univalent ions (Na+, K+, Cl-) to pass. Variation and
adjustment of the parameters for nanofiltration are well within the
knowledge of the skilled artisan, merely depending on the volume of
the solution and of the equipment used. This fraction will be
further referred to as FHMW or FLMW depending on whether the fucans
fraction is native or depolymerized, respectively.
Preparation of Galactans
[0140] In a specific embodiment, galactans of the present invention
are obtainable from Asparagopsis armata, cultured according to the
patent application EP-0733636 A1. This patented cultivation
technique is based on the principles of vegetative algal
propagation, and the use of a special type of rope. This unique
rope is seeded with segments of wild thalli which then proliferate
by vegetative propagation/fragmentation and significantly increase
in biomass.
[0141] Fresh, frozen or dried clean algae can be transformed into a
fine powder. Fresh and frozen algae are chopped to obtain particle
size smaller than about 8 mm, preferably between about 0.5 mm and
about 8 mm, most preferably between about 2 and about 5 mm as
described above.
Native Galactans (i.e. Which Naturally Contain Minerals)
[0142] Galactans are extracted from the chopped algae by an
adequate volume of water-based solution. The volume of solution
used can be increased without bearing any deleterious effect on the
yield of recovery of valuable components. A small volume is
preferred since it is more convenient to manipulate than larger
volumes. Water is then purified by inverse osmosis and multiple
filtrations down to 0.1 micrometer filter. Many aqueous solutions
(containing salts, for example) could be used in lieu of water.
When recovery of a plurality of hydrosoluble activities is
contemplated, working at a near neutral pH (6.0 to 8.0) and
non-denaturing conditions is preferred to avoid degradation or
denaturation of some of the active components. For the sake of
clarity, any extraction medium that is compatible with the
preservation of biologically active galactan components is within
the scope of this invention. Therefore, performing the extraction
in pure water is preferred. Other preferred embodiments include
those wherein salts and/or chaotropic agents are added to the water
prior to or during extraction.
[0143] The extraction is performed at about 50 to about 90 degrees
Celsius, preferably ranging between about 60 to about 80 degrees
Celsius. The extraction time is about 1 to 8 hours, preferably
between about 4 and 6 hours. The speed of the agitation as well as
the volume of aqueous solution may influence both time and yield of
extraction.
[0144] The supernatant is then separated from the pellet. The
separation can be quickly performed by filtration (with mesh of
about 50 micrometer), centrifugation, or with a commercial
decanter. Variation and adjustment of the separation parameters are
well within the knowledge of the skilled artisan, merely depending
on the volume of homogenate and of the equipment used.
[0145] The resulting supernatant is then clarified to get rid of
fine suspension susceptible to affect the performance of the
process. The clarification was performed at a temperature ranging
from about 50 to about 70 degrees Celsius, preferably at about 60
degrees Celsius.
[0146] The clarified material is then cooled down at a temperature
ranging from about 30 to about 50 degrees Celsius, preferably from
about 40 degrees Celsius.
[0147] The clarified material is then acidified at a pH ranging
from about 3 to about 5, preferably about 4.5. For example, the
acidification could be performed in presence of sulfuric acid or
hydrochloric acid. The acidified materiel is then clarified as
described above.
[0148] The acidified materiel is then depigmented by
microfiltration and then clarified as described above. Variation
and adjustment of the parameters for microfiltration are well
within the knowledge of the skilled artisan, merely depending on
the volume of the solution and of the equipment used. The solution
is then neutralized.
[0149] The depigmented solution is then concentrated. This step can
be performed by different means including but not limited to:
dialysis, chromatography (adsorption, ionic exchange, gel
filtration), electrophoresis, ultrafiltration, ultra centrifugation
with zonal density gradients, adsorption and extraction.
[0150] In a specific embodiment, the solution is ultrafiltered at
about 4 degrees Celsius to about 25 degrees Celsius (although
temperature could be increased to about 40 degrees Celsius) on a
tangential flow filtration column having a membrane of a porosity
of about 10 kDa. The obtained fraction is then dialyzed which
allows a first fractionated extract to be obtained, comprising
hydrosoluble galactans of an average molecular weight generally
higher than the membrane cut off taking into account the
variability inherent to the membrane pores and to the conformation
of the molecules in the extraction solvent higher than the membrane
cut off.
[0151] The galactans present in the solution are then precipitated
in the presence of alcohol and the precipitate is dried. Variation
and adjustment of the parameters for alcohol precipitation and
drying are well within the knowledge of the skilled artisan, merely
depending on the volume of the solution and of the equipment used.
This galactan-enriched fraction will be further referred to as the
mineralized galactan fraction (GHMW-M).
[0152] Molecular analysis of galactans fractions obtained from red
algae revealed that galactose constitutes between about 15 and 95%,
preferably between about 20 and 75%, and more preferably between
about 30 and 40% of the dry weight of the galactans. Moreover, 1 to
10%, and more preferably 1-5% of the dry weight is composed of
uronic acid. These galactans are richly sulfated, since about 1 to
60% and more preferably about 20 to 35% of the dry weight is
composed of sulfates (SO.sub.4) radical.
Enzymatic Dosage of D-Galactose
[0153] The dosage of D-galactose is performed with a dosage kit
(test combining lactose/D-galactose, Boehringer Mannheim n.
E0176303). D-galactose is oxydized at pH 8,6 by beta-galactose
dehydrogenase (Gal-DH), in the presence of NAD (nicotinamide
adenine Dinucleotide).
D-galactose+NAD.sup.+.fwdarw.D-galactonic acid+NADH+H.sup.+
[0154] The reaction being stoechiometric, the quantity of NADH
formed is equivalent to the quantity of oxydized D-galactose. It is
followed by a measure of absorbance at 340 nm. A standard curve is
realized from D-galactose. 15 mg of polysaccharides are hydrolized
by 1 mL of trifluoroacetic acid 2 M during 90 minutes at
120.degree. C. 20 mg of D-galactose are also hydrolized in the same
conditions. Hydrolysates are evaporated three times at
Rotavapor.TM., using each time in distilled water and redissolved
in 10 ml of milli-Q.TM. for the polysaccharide or in 100 ml of
milli-Q water for the D-galactose. The pH of these two solutions is
then adjusted at 6,6 (i.e. pH of the citrate buffer of the dosage
kit) with the assistance of sodium hydroxide 0.05 M. 100 and 200
.mu.L of the polysaccharides solution at a concentration of 1.5
mgmL.sup.-1 are placed in glass tubes and assayed according to the
manufacturer's instructions. 100 to 500 .mu.L of the solution of
D-galactose at a final concentration of 0.2 gL.sup.-1 are used to
produce the standard curve. The absorbance is read after 45 minutes
at a wavelength of 340 nm, and the D-galactose amount found in the
polysaccharide is measured in light of the standard curve. Knowing
the total quantity of galactose contained in the polysaccharide
(dosed in the form of alditols acetates.) the L-galactose amount in
the polysaccharides may be derived.
Determination of Sulphate Content and Uronic Acid Content of
Extracted Fucans and Galactans
[0155] Total sugars and uronic acids were quantified on each
polysaccharide using phenol-sulfuric acid method (Dubois et al.,
1956) using glucose as standard and Blumenkrantz method
(Blumenkrantz and Asboe-Hansen, 1973) with glucuronic acid as
standard and 3-phenyl-phenol as reagent. Absorbance measurements
were realised in triplicate. Microplate reader (Molecular devices,
Ottawa, ON, Canada) was used for total sugars analysis and HP-8453
spectrophotometer (Hewlett Packard, Mississauga, ON, Canada)
equipped with UV-visible ChemStation.TM. software for uronic acids.
Minerals (Ca.sup.2+, MG.sup.2+, Na.sup.+ and S) were quantified by
ICP-OES (Inductively Coupled Plasma-Optical Emission Spectroscopy)
with model Optima.TM. 4300 DV from Perkin-Elmer (USA) equipped with
Winlab32.TM. software. Sulphates content was deduced from the
amount of sulphur determined by ICP described by this relation: %
sulphate group=3.22.times.S (Roger et al., 2004).
Estimation of the Molecular Weight of Polysaccharides Formed by
HPSEC (High Performance Size Exclusion Chromatography)
[0156] The molecular mass of oligosaccharides formed by radicalar
depolymerization is determined by high performance steric
exclusion. It allows to evaluate the number average molecular
weight (Mn) each oligosaccharides, molecular weight at the peak
(Mp), and the polydispersity indicia (Ip) (=Mw/Mn) which
characterize the polysaccharides chains homogeneity. The
calibration is obtained with pullulan standards (Nardella et al.,
1996). Pullulans are glucanes, namely neutral polysaccharides,
while oligosaccharides from Asparagopsis armata are highly
sulfated. This is the reason why the estimated mass of Asparagopsis
armata oligosaccharides can be assimilated to equivalent pullulans
masses but does not reflect the real mass of these
oligosaccharides. This method is also very useful to compare
various fractions obtained during radicalar depolymerization
performed in different conditions. The protocol used is described
in Mulloy et al. (1997). The TSK.TM. G3000 SW-XL and TSK.TM. G2000
SW-XL columns in series were used. The eluting agent is a solution
of ammonium acetate at o,1 M, and the flow rate applied is of 0.5
mLmin.sup.-1. The detection is made by refractometry
(refractomonitor W, LDC, Analytical, Stone, Saffs., UK). The data
treatment is made by a software from Polymer Laboratories, Church
Stretton, UK.
[0157] In accordance with a specific embodiment of the present
invention, the galactans fraction was characterized by the method
described above. According to such method, galactose constituted
about 37% of the dry weight of the fraction, uronic acid
constituted about 3% of the dry weight of the fraction, and
sulfates constituted about 27% of the dry weight of the fraction.
The average molecular weight of these galactans was higher than
about 100 kDa and in more specific embodiments, the Mw was about
350 kDa (obtained from a pullulan equivalent).
Depolymerized Galactans
[0158] The galactans could be used entirely or partly in their
depolymerized form. In a specific embodiment, the galactans'
depolymerization is performed by acidic hydrolysis. Native
galactans are acidified to a pH of about 0.5 to about 2.0,
preferably to about 0.5 to about 1.0. The acidified solution is
incubated at temperature of about 55 to about 70 degree C. for
about 4 hours. Depolymerization may be achieved by different
methods including but not limited to: ultrasounds, UV radiations,
ozonolysis, chemical, radical, or enzymatic hydrolysis, and high
pressure and temperature.
[0159] The solution is then neutralized and then
concentrated/purified on a 2 kDa ultrafiltration followed by a
dialysis.
[0160] The galactans present in the solution are then precipitated
in the presence of alcohol and the precipitate is dried. Variation
and adjustment of the parameters for alcohol precipitation and
drying are well within the knowledge of the skilled artisan, merely
depending on the volume of the solution and of the equipment used.
This fraction will be further referred to as the mineralized
depolymerized galactans fraction (GLMW-M).
Demineralized Galactans
[0161] The galactans fractions could also be used entirely or
partly in their demineralised form. Demineralization could be
industrially performed such as with but not limited to
nanofiltration as described above. This fraction will be further
referred to as GHMW.
[0162] The present invention is illustrated in further details by
the following non-limiting examples.
Example 1
Preparation of Fucans
[0163] Fucans were obtained from Ascophyllum nodosum. Fresh and
frozen algae were chopped to obtain a particle size between 2 and 5
mm.
[0164] Fucans were extracted from the chopped algae in a
water-based solution.
[0165] The extraction of fucans was performed at a temperature
varying from 80 to 95 degrees Celsius for a time of 10 to 16
hours.
[0166] The supernatant was then separated from the pellet with a
commercial decanter.
[0167] The resulting supernatant was then clarified to get rid of
fine suspension susceptible to affect the performance of the
process. The clarification was performed at 60 degrees Celsius
[0168] The clarified material was then cooled down at 25 degrees
Celsius and was then acidified at pH 2 to precipitate proteins and
alginates. The acidified materiel was then clarified and
neutralized as described above.
[0169] The neutralized solution was then purified/concentrated by
ultrafiltration and dialysis to obtain a fraction containing fucans
with average molecular weight higher than 100 kDa to obtain a
naturally mineralized native fucans fraction (FHMW-M).
Depolymerized Fucans
[0170] The fucans depolymerization was performed by acidic
hydrolysis. Native fucans (FHMW-M) were acidified to a pH ranging
from 0.5 to 1.0. The acidified solution was incubated at 65 degrees
Celsius for 4 hours. The solution was then neutralized and
concentrated as described above.
[0171] The fucans present in the solution were then precipitated in
the presence of ethanol and the precipitate was dried to obtain a
mineralized depolymerized fucan fraction (FLMW-M). Fucans average
molecular weight was generally ranging from about 5 kDa to about 25
kDa.
Demineralized Fucans
[0172] Demineralization was performed by nanofiltration to obtain
the FHMW or FLMW fraction depending on whether the fucans fraction
was native or depolymerized.
Molecular Analysis of Fucans
[0173] The molecular analysis of the FHMW-M fraction revealed that
fucose constituted between 25 and 35% of the dry weight. Moreover,
12 to 20% of the dry weight was composed of uronic acid. These
fucans are richly sulfated, since 15 to 25% of the dry weight was
composed of sulfates (SO.sub.4) radical.
Example 2
Preparation of Galactans
[0174] Galactans were obtained from Asparagopsis armata, which were
cultured according to the patent application EP-0733636 A1 as
described above.
[0175] Fresh and frozen algae were chopped to obtain particle size
smaller between 0.5 and 2 mm.
[0176] Galactans were extracted in water at pH 8 and at a
temperature ranging from 60 to 80 degrees Celsius for about 4 to 6
hours.
[0177] The supernatant was then separated from the pellet with a
commercial decanter. The resulting supernatant was then clarified
to get rid of fine suspension susceptible to affect the performance
of the process. The clarification was performed at 60 degrees
Celsius The clarified material was then cooled down at 40 degrees
Celsius and the pH was adjusted at 4.5.
[0178] The acidified material was then depigmented by tangential
microfiltration (1.2 micrometer).
[0179] The depigmented solution was then concentrated/purified
through ultrafiltration with a tangential flow filtration column
having a membrane of a porosity of 10 kDa. The obtained fraction
was then dialyzed so as to obtain a fraction containing molecules
of higher than about 100 kDa. The galactans present in the solution
were then precipitated in presence of alcohol and the precipitate
was dried to obtain a native galactans fraction that is naturally
mineralized (GHMW-M).
Demineralized Galactans
[0180] The galactans fractions could be used and prepared entirely
or partly in their demineralised form. Demineralisation was
performed by nanofiltration as described above to obtain a
mineralized demineralized galactans fraction (GHMW).
Molecular Analysis of Galactans
[0181] The molecular analysis of the GHMW fraction obtained from
these red algae revealed that galactose constituted about 37% of
the dry weight. Moreover, 3% of the dry weight was composed of
uronic acid. These galactans were richly sulfated, since about 27%
of the dry weight was composed of sulfates (SO.sub.4) radical.
Example 3
Description of Fucans and Galactans Fractions
[0182] The different fractions prepared and tested either
separately or in combinations are summarized in Table 3 below.
TABLE-US-00003 TABLE 3 DESCRIPTION OF THE DIFFERENT FUCAN AND
GALACTAN FRACTIONS FHMW This preparation contains native
sulfated-fucans, having an average molecular weight ranging from
about 0.1 kDa to about 100 kDa. This fraction was demineralized by
nanofiltration. FLMW Preparation containing depolymerized
sulfated-fucans, which was demineralized by nanofiltration. The
average molecular weight of these fucans is ranging from about 5
kDa to about 25 kDa. This preparation was demineralized by
nanofiltration. FLMW-M Preparation containing depolymerized
sulfated-fucans which was not demineralized. The average molecular
weight of these fucans is ranging from about 5 kDa to about 25 kDa.
FHMW-M Preparation containing native sulfated-fucans (i.e. before
concentration and nanofiltration). The average molecular weight of
the fucans present in this fraction is from about 0.1 kDa to about
100 kDa. GHMW Preparation containing native galactans. The average
molecular weight of these galactans is higher than about 100 kDa.
This preparation was demineralized by nanofiltration. GHMW-M
Preparation containing native galactans, which was not
demineralized. The average molecular weight is higher than about
100 kDa.
Example 4
Hen's Egg Chorioallantoic Membrane Test for Irritation Potential of
Fucans/Galactans Mixtures
[0183] The Hen's Egg Test (HET) is a rapid and sensitive toxicity
test and can give information on mucous-membrane irritation
potencies of chemical substances. Testing with incubated hen's eggs
is a borderline case between in vivo and in vitro systems. A
specific score and classification scheme was developed for the
HET.
[0184] The ocular irritancy potential of a composition containing
50 mg/ml of FLMW and 5 mg/ml of GHMW was tested at 0.2% (0.11
mg/ml) was performed as previously described by Luepke and Kemper
(Luepke N P & Kemper F H (1985) Hen's egg chorioallantoic
membrane test for irritation potential Food Chem. Toxicol. 1985
(2):287-291). Briefly, the tested compositions were applied to the
chorio allantoic membranes for 20 seconds then rinsed off with NaCl
0.9% (w/v). Injection, hemorrhage and coagulation parameters were
scored during 5 minutes after treatment. The control preparations
containing lauryl sulfo betain at 3.2%, 0.4% were classified as
irritant while the 0.05% preparation was classified as practically
non irritant, for the chorioallantoic membrane of chicken egg. The
tested composition was classified as non irritant for the
chorioallantoic membrane of chicken egg.
Example 5
[0185] Effect of Polysaccharides Compositions on IL-8, PGE2 and
VEGF Levels in Human Keratinocytes in Vitro
[0186] Recent publications have shown that PMA activation led
normal keratinocytes to produce increased amounts of interleukin-8
(IL-8), a mediator of inflammation (Chabot-Fletcher et al. (1994)
J. Invest. Dermatol. 103:509-515). Moreover, psoriatic
keratinocytes produce very high amounts of IL-8, which further
promote neovascularization in psoriatic plaques (Nickoloff et al.
(1994) Am. J. Pathol. 144:820-828).
[0187] As indicated above, VEGF is a multifunctional agent. It is a
potent vascular permeabilization agent that renders the endothelial
tissue hyperpermeable, leading to the extravasation of plasma
proteins and leukocytes migration to the inflammatory site. It is
also a multifunctional angiogenic growth factor that stimulates the
endothelial cells to proliferate and to migrate.
[0188] Inhibition of the production of IL-8 and VEGF in
PMA-activated keratinocytes was taken to indicate herein that
tested polysaccharide compositions have anti-inflammatory and
anti-extravasation activities.
[0189] Human keratinocytes (NCTC2544) were cultured in DMEM
containing 5.5 mM glucose, 2 mM L-glutamine, and 10% FCS. Cells
were pre-treated with various fucan/galactan polysaccharides
compositions of the invention (0.06-2.4 mg/ml, unless otherwise
indicated) during 24 hours. Afterward, the culture media were
replaced for one containing the polysaccharides compositions in the
presence of 0.1 .mu.g/ml of Phorbol 12-Myristate 13-acetate (PMA).
Twenty four hours later the culture media were collected for
further analysis. VEGF, IL-8, PGE.sub.2 contents in the culture
media were assayed by ELISA using commercial kit from Diaclone inc.
and R&D systems inc.
TABLE-US-00004 TABLE 4 EFFECT OF VARIOUS POLYSACCHARIDE
COMPOSITIONS ON IL-8, PGE.sub.2 AND VEGF LEVELS IN HUMAN
KERATINOCYTES IN VITRO Effect on PMA-treated cells IL-8 PGE.sub.2
VEGF (% (% (% Treatment Concentration inhibition) inhibition)
inhibition) FHMW 2.4 mg/ml 6 62** 74** 1.2 mg/ml 7 63** 71** 0.6
mg/ml 4 59** 71** FLMW 2.4 mg/ml 0 64** 73** 1.2 mg/ml 18** 66**
75** 0.6 mg/ml 22** 64** 74** GHMW 0.24 mg/ml 9 49** 40** 0.12
mg/ml 9 45** 32** 0.06 mg/ml 5 34** 35** FHMW/GHMW 1.96 mg/ml 0
62** 71** (40/1 w/w) 0.984 mg/ml 15* 60** 70** 0.492 mg/ml 15 66**
69** FHMW/GHMW 1.32 mg/ml 8 57** 72** (10/1 w/w) 0.66 mg/ml 11 62**
70** 0.33 mg/ml 30** 65** 71** FHMW/GHMW 0.672 mg/ml 15** 59** 73**
(2.5/1 w/w) 0.336 mg/ml 16** 64** 71** 0.168 mg/ml 24** 64** 71**
FLMW/GHMW 1.96 mg/ml 28** 67** 74** (40/1 w/w) 0.984 mg/ml 34**
70** 69** 0.492 mg/ml 42** 69** 73** FLMW/GHMW 1.32 mg/ml 15 69**
81** (10/1 w/w) 0.66 mg/ml 13 70** 79** 0.33 mg/ml 26** 75** 80**
FLMW/GHMW 0.672 mg/ml 6 68** 79** (2.5/1 w/w) 0.336 mg/ml 22** 72**
76** 0.168 mg/ml 26** 71** 76** FHMW-M 4 mg/ml 0 97** 39** 2 mg/ml
49** 98** 21** 1 mg/ml 40** 98** 14 FLMW-M 4 mg/ml 30** 64 73** 2
mg/ml 38** 70 73** 1 mg/ml 45** 68 72** *P < 0.05 **P <
0.01
[0190] The results indicated that PMA strongly upregulates the
production of VEGF in keratinocytes. Indomethacin, a non-selective
cyclooxygenase inhibitor, partially inhibits PMA-induced VEGF
expression. This suggests that the PMA-induced VEGF production is
not solely dependent on the cyclooxygenase pathway and that other
mediators might be involved. This is in line with what has been
observed by Trompezinski and collaborators (Trompezinski S, Pernet
I, Schmitt D, Viae J. Inflamm Res. 2001; (50):422-427).
[0191] The results also showed that fucans/galactans compositions
of the present invention are able to significantly inhibit
PMA-induced keratinocyte activation of VEGF and IL-8. For example,
a concentration of 0.66 mg/ml of the fucans/galactans (10/1)
reduces by 79% (p<0.01) the VEGF production (see FIG. 1).
Without being so limited, these results do not exclude the
hypothesis that the action of this composition aims at more than
one stress-induced signalling pathway in skin cells. Fucans and
galactans compositions were also tested individually at a
concentration equivalent to that found in the fucans/galactans
combination tested. Fucans are more effective than galactans in
reducing the PMA-induced production of VEGF. However, there is an
unexpected synergistic effect of the fucans/galactans composition
in inhibiting VEGF (see FIG. 1). Indeed, there is a significant
difference (p<0.05) between the effect of the fucans/galactans
composition and the effect of the individual sulfated-fucans or
galactans in inhibiting the PMA-induced increase in VEGF. Similar
results were observed with IL-8 (see Table 4).
[0192] Results also showed that PGE.sub.2 production is highly
increased in keratinocytes upon a stress induced by the
pro-inflammatory agent PMA. Indomethacin, a non-selective
cyclooxygenase inhibitor, displays a complete inhibition of the
PMA-induced PGE.sub.2 expression.
[0193] Fucans/galactans compositions of the present invention are
able to significantly inhibit PGE.sub.2 release from PMA-treated
keratinocytes. For example, a concentration of 0.66 mg/ml of the
fucans/galactans (10/1) composition significantly reduces (70%
reduction, see FIG. 2) PGE.sub.2 production. The effect of the
demineralised compositions was not higher than the sulfated-fucans
and galactans alone. The inhibition by FHMW-M was however close to
100%. Mineralized molecules may thus substitute for at least a part
of the demineralized polysaccharides and provide better
compositions.
[0194] Results obtained with human keratinocytes demonstrate that
the fucans/galactans composition inhibits the induction of VEGF and
PGE.sub.2 in human keratinocytes. PGE.sub.2 is a prostaglandin that
mediates microcapillary dilatation (Rhodes L E, Belgi G, Parslew R,
McLoughlin L, Clough G F, Friedmann P S. J Invest Dermatol. 2001
October; 117(4):880-5: Rhodes L E, Belgi G, Parslew R, McLoughlin
L, Clough G F, Friedmann P S. J Invest Dermatol. 2001 October;
117(4):880-5) and VEGF is a growth factor that plays a primary role
in the phenomenon of microcapillary hyperpermeability (Brauchle M,
Funk J O, Kind P, Werner S. J Biol chem. 1996 Sep. 6;
271(36):21793-7; Dvorak H F, Brown L F, Dvorak A M. Am J Pathol.
1995 May; 146(5):1029-39). Even though the VEGF and PGE.sub.2
pathways may be interrelated especially when up-regulated by
various stresses (Harada S, Nagy J A, Sullivan K A, Thomas K A,
Endo N, Rodan G A, Rodan S B. J clin Invest. 1994 June;
93(6):2490-6; Cheng T, Cao W, Wen R, Steinberg R H, LaVail M M.
Invest Ophthalmol Vis Sci. 1998 March; 39(3):581-91; Trompezinski
S, Pernet I, Schmitt D, Viae J. Inflamm Res. 2001; (50):422-427),
there are indications that, at least in part, they obey to
independent and different upstream signal transduction pathways
(Trompezinski S, Pernet I, Schmitt D, Viae J. Inflamm Res. 2001;
(50):422-427, Bachelor M A, Bowden G T. Seminars in Cancer Biology.
2004; 14:131-138). Those observations demonstrate the need to
simultaneously inhibit VEGF and PGE.sub.2 pathways to prevent
stress-induced skin photo-damage.
Example 6
Effect of Polysaccharides Compositions on Human Keratinocytes
Viability In Vitro
[0195] Human keratinocytes (NCTC2544) were cultured in DMEM
containing 5.5 mM glucose, 2 mM L-glutamine, and 10% FCS. Cells
were pre-treated with polysaccharides compositions (0.06-2.4 mg/ml,
unless otherwise indicated) during 24 hours. Afterward, the culture
media were replaced for one containing the polysaccharides
compositions in the presence of 0.1 .mu.g/ml of Phorbol
12-Myristate 13-acetate (PMA). Cell's viability was determined by
measuring the activity of mitochondrial dehydrogenase (MTT Cell
Proliferation Assay). The MTT Cell Proliferation Assay measures the
cell proliferation rate and conversely, when metabolic events lead
to apoptosis or necrosis, the reduction in cell viability. The
assay is based upon the capacity of the mitochondrial dehydrogenase
to reduce the yellow tetrazolium MTT
(3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide), to
generate reducing equivalents such as NADH and NADPH. The resulting
intracellular purple formazan can be solubilized and quantified by
spectrophotometric means.
TABLE-US-00005 TABLE 5 EFFECT OF FUCANS AND GALACTANS PREPARATIONS
ON HUMAN KERATINOCYTES VIABILITY IN VITRO Polysaccharide Cell's
viability preparation Concentration % of MTT conversion FHMW 2.4
mg/ml 96 1.2 mg/ml 98 0.6 mg/ml 98 FLMW 2.4 mg/ml 101 1.2 mg/ml 98
0.6 mg/ml 92 GHMW 0.24 mg/ml 78 0.12 mg/ml 77 0.06 mg/ml 71
FHMW/GHMW (40/1 w/w) 1.96 mg/ml 101 0.984 mg/ml 99 0.492 mg/ml 100
FHMW/GHMW (10/1 w/w) 1.32 mg/ml 107 0.66 mg/ml 106 0.33 mg/ml 100
FHMW/GHMW 0.672 mg/ml 104 (2.5/1 w/w) 0.336 mg/ml 109 0.168 mg/ml
105 FLMW/GHMW (40/1 w/w) 1.96 mg/ml 105 0.984 mg/ml 98 0.492 mg/ml
99 FLMW/GHMW (10/1 w/w) 1.32 mg/ml 103 0.66 mg/ml 111 0.33 mg/ml
103 FLMW/GHMW 0.672 mg/ml 105 (2.5/1 w/w) 0.336 mg/ml 106 0.168
mg/ml 102 FHMW-M 4% 82 2% 86 1% 90 FLMW-M 4% 85 2% 97 1% 100 *P
< 0.05 **P < 0.01
[0196] Results showed that high molecular weight galactans slightly
reduced the keratinocytes viability, while no difference in
viability was observed in the fucans treated-cells. For example,
0.6 mg/ml of high molecular weigh galactans reduces the cell
viability by 30% while no viability alteration was observed at
concentrations of fucans as high as 2.4 mg/ml. Surprisingly, the
effect of galactans on keratinocyte viability was entirely
prevented by the presence of fucans.
Example 7
Clinical Efficacy of the Fucans/Galactans Compositions in
Maintaining the Microcapillary Integrity upon Skin UV Exposure In
Vivo
[0197] Increased dilation and the subsequent increase in
microcapillary circulation is part of the thermoregulative system
(Charkoudian, 2003; Marszalek, 1996; Kellogg, 1993). In the skin,
this physiological reaction is under the control of so-called
neurogenic transmitters (Kellogg, 2005, 2003 and 1998; Bennett,
2003). However, skin microcapillary dilation also occurs in
response to various stresses. For instance, an increase in skin
microcirculation in response to UV has also been demonstrated
(Benrath, 2001; Terui, 2001; Nose, 1993; Frodin et al., 1988). This
phenomenon of dilation not only correlates with an increase in
microcapillary circulation but also with a plasma protein leakage
and leukocytes efflux (Holzer, 1998).
[0198] The measurement of skin microcapillary circulation has been
widely used to assess the cutaneous response to physiological and
pathophysiological conditions such as in reaction to allergens,
vasodilators and in cases of atypical dermatitis, rosacea and
psoriasis (Hee Chul Eun, 1995). Measurement of skin
microcirculation can be more sensitive than the visualization of
erythema (Wahlberg, 1992) and can thus be utilized to assess
changes in the skin microcapillary integrity.
[0199] A device was used to measure cutaneous microcirculation. The
skin thermal conductivity (K) is directly proportional to the
superficial cutaneous microcirculation and is based on the capacity
with which the heat is transported by skin microcirculation. In the
case of biological measurements, there is a linear relationship
between cutaneous microcirculation and thermal conductivity. This
conductivity is measured with a specific thermal device
(exemplified by Hematron.TM.) which is applied directly onto the
skin. Microcapillary dilation and the resulting increase of blood
circulation will increase the skin thermal conductivity.
[0200] Clinical Study--Preventive Action Against UV-Exposure:
[0201] Subjects applied either the placebo or a formulation
containing 5% of 5.5 mg/ml of FLMW/GHMW (10/1) composition, twice
daily for a period of 7 days on separate skin areas (day -7 to Day
0). One skin area was left as is, with no application of products
as a control. On Day 0, the Hematron.TM. was used to measure the
baseline skin microcapillary circulation. On the same day, subjects
were exposed to 1 MED of UV (Day 0, T0h). Five hours after UV
exposure (Day 0, T5h), skin microcirculation was again measured on
all skin areas.
[0202] As illustrated in FIG. 3, results show that a topical
formulation containing 5% of 5.5 mg/ml of FLMW/GHMW (10/1) in a
topically acceptable carrier, namely a cream, was able to reduce
the UV-induced increase skin microcirculation to a value close to
what was observed in a non-irradiated skin area (baseline). The
present formulation demonstrated an efficacy of 67% when applied as
a preventive care.
[0203] Clinical Study--Post UV Exposure Action:
[0204] Subjects were first exposed to 1 MED of UV readily on Day 0.
Five hours after UV exposure (Day 0, T5h), the Hematron.TM. was
used to assess the UV-induced increase in skin microcirculation.
Subjects then applied either the placebo or a formulation
containing 5% of 5.5 mg/ml of FLMW/GHMW (80/20) composition on
irradiated areas (Day 0, T5h post UV). One skin area was left as
is, with no application of products as a control. Two hours later,
(Day 0, T7h) skin microcirculation was again measured on all skin
areas.
[0205] As illustrated in FIG. 4, results show that a formulation
containing 5% of 5.5 mg/ml of FLMW/GHMW (10/1) composition was able
to reduce the UV-induced increase in skin microcirculation to a
value close to what was observed for a non-irradiated skin area
(baseline). The present formulation demonstrated an efficacy of 74%
when applied post UV exposure.
Example 8
TABLE-US-00006 [0206] Fucans/Galactans Formulations Formulation 1
Demineralized water, Sepigel 305, Lanol 1688, Abil 8839, Irgasan
DP300, Ethanol, Hydroxan .RTM. CH, Phenonip, Fucan/Galactan
composition 0.1-99.9% of 5.5 mg/ml, JFL 544/2. Formulation 2
Arlacel 165 .TM., Arlacel 60 .TM., Tween 60 .TM., Sipol C16P .TM.,
Crodamol OP, Miglyol 840 .TM., Demineralized water, Sequestrene NA4
.TM., Phenonip, Sepigel 305 .TM., Fucan/Galactan composition
0.1-99.9% of 5.5 mg/ml. Formulation 3 Arlacel P135 .TM., Amerchol
L10 .TM., Amerlate P .TM., Isohexadecane, Magnesium Stearate,
Parsol MCX .TM., Parsol 1789 .TM., LNST .TM. 98, Nipastat .TM.,
Demineralized water, Eospoly UV cristal HL .TM., Sequestrene NA4
.TM., Cire DC 2501, Demineralized water, Magnesium Sulphate, Abil
8839 .TM., Micropearl M100 .TM., Fucan/Galactan composition
0.1-99.9% of 5.5 mg/ml, Lanachrys .TM., Joyty Concentrate.
Formulation 4 Demineralized water, Carbopol Ultrez 10 .TM., Amigel
.TM. (sol. 1% Aq. Phenonip, Demineralized water, TEA, Hydralphatine
.TM. 3P, Phenonip .TM., Pure Ethanol, Abil 8839 .TM., Joyty
Concentrate, LRI Solubilizer, Fucan/Galactan composition 0.1-99.9%
of 5.5 mg/ml. Formulation 5 Cetyl alcohol (and) Glyceryl stearate
(and) PEG-75 .TM. stearate (and) Ceteth-20 .TM. (and) Steareth-20
.TM., Mineral oil (and) Lanolin alcohol, Isopropyl lanolate,
Mineral oil (and) Prunus Armeniaca (Apricot) Kernel oil (and)
Calendula officinalis flower extract, LNST .TM. 98, Demineralized
water, Carbomer .TM. (2% aqueous solution), Red 33 (10% aqueous
solution), Blue 1 (10% aqueous solution), Cyclopentasiloxane (and)
cyclohexasiloxane, Demineralized water, Triethanolamine,
Phenoxyethanol (and) Propylparaben (and) Butylparaben, Glycerin,
Aluminium starch octenyl succinate, Fucan/Galactan composition
0.1-99.9% of 5.5 mg/ml, Fragrance. Formulation 6 Purified Water,
Glycerin, Nipagin M .TM., TEA 99%, Irgasan DP300 .TM., Promulgen D
.TM., Tegosoft OP .TM., Blandol Mineral Oil, Shea Butter, GMS-SE,
Lipovol Soy .TM., Vitamin E Acetate, Nipasol M .TM., Fragrance
700F48, Fucan/Galactan composition 0.1-99.9% of 5.5 mg/ml.
Formulation 7 Purified Water, Propylene Glycol, Magnesium Ascorbyl
Phosphate, Nipagin M .TM., Sodium Citrate, Irgasan DP300 .TM.,
Promulgen D .TM., Tegosoft OP .TM., Blandol Mineral Oil, Shea
Butter, GMS-SE, Lipovol Soy .TM., Vitamin E Acetate, Nipasol M
.TM., Fragrance 700F47, Fucan/Galactan composition 0.1-99.9% of 5.5
mg/ml.
Example 9
TABLE-US-00007 [0207] Sensitive skin cream Ingredients Wt %
Demineralized water QSP100 Xanthan Gum 0.15 Phenoxyethanol (and)
Methylparaben (and) Butylparaben 0.60 (and) Ethylparaben (and)
Propylparaben Chlorphenesin 0.20 Glycerin 3.00 Cetearyl Alcohol
(and) Cetearyl Glucoside 4.00 Caprylic/Capric Triglyceride 2.00
Jojoba Esters 0.50 Squalane 3.00 Shea Butter 0.50 Wheat Germ oil
1.00 Onager oil 1.00 Macadamia Ternifolia Seed oil 1.00 PEG-8 (and)
Tocopherol (and) Ascorbyl Palmitate (and) 0.10 Ascorbic Acid (and)
Citric Acid Tocopheryl Acetate 0.20 Aluminium Starch
Octenyl-Succinate 1.50 Aldavine .TM. (fucans/galactans 10/1
0.1-99.9%) 1.00-5.00 Fragrance 0.30 Citric acid 0.04
Example 10
TABLE-US-00008 [0208] Firming cream Ingredients Wt % Demineralized
water QSP10 Glycerin 3.00 Phenoxyethanol (and) Methylparaben (and)
Butylparaben 0.60 (and) Ethylparaben (and) Propylparaben
Chlorphenesin 0.20 Cetearyl Alcohol (and) Cetearyl Glucoside 6.00
Caprylic/Capric Triglyceride 5.00 Squalane 3.00 Dimethicone 1.00
Shea Butter 2.00 Cetearyl Alcohol 1.00 Hydrogenated coco-glyceride
3.00 Cetyl Acetate (and) Acetylated lanaolin Alcohol 0.50 Cetyl
Alcohol 2.00 PEG-8 (and) Tocopherol (and) Ascorbyl Palmitate (and)
0.10 Ascorbic acid (and) Citric acid Aldavine .TM.
(fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Polyacrylamide (and)
C13-14 isoparaffin (and) Laureth-7 0.30 Fragrance 0.30
Example 11
TABLE-US-00009 [0209] Firming serum-lotion Ingredients Wt %
Demineralized water QSP100 Hydroxyethylcellulose 0.40 Glycerin 3.00
Phenoxyethanol 0.40 Benzyl alcohol 0.60 Carbomer 0.05 Sodium
Hydroxyde (10% aqueous solution) 0.05 Ethylhexyl Methoxycinnamate
(and) Butyl 0.50 Methoxydibenzoylmethane (and) Ethylhexyl
Salicylate (and) PPG-26-Buteth-26 (and) PEG-40-Hydrogenated Castor
oil Bis-PEG-18 Methyl Ether Dimethyl Silane 2.00 Aldavine .TM.
(fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Fragrance 0.10
PPG-26-Buteth-26 (and) PEG-40-Hydrogenated Castor oil 0.20 Blue 1
(0.1% aqueous solution) 0.09
Example 12
TABLE-US-00010 [0210] Anti-age mask Ingredients Wt % Demineralized
water QSP100 Glycerin 3.00 Phenoxyethanol (and) Methylparaben (and)
Butylparaben 0.60 (and) Ethylparaben (and) Propylparaben
Chlorphenesin 0.20 Cetearyl Alcohol (and) Cetearyl Glucoside 6.00
Caprylic/Capric Triglyceride 5.00 Squalane 3.00 Dimethicone 1.00
Shea Butter 2.00 Cetearyl Alcohol 1.00 Hydrogenated coco-glyceride
3.00 Cetyl Acetate (and) Acetylated lanolin Alcohol 0.50 Cetyl
Alcohol 2.00 Kaolin 5.50 PEG-8 (and) Tocopherol (and) Ascorbyl
Palmitate (and) 0.10 Ascorbic acid (and) Citric acid Aldavine .TM.
(fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Polyacrylamide (and)
C13-14 isoparaffin (and) Laureth-7 0.30 Fragrance 0.30 Citric Acid
(10% aqueous solution) 0.07
Example 13
TABLE-US-00011 [0211] After-sun cream Ingredients Wt %
Demineralized water QSP100 Ethylhexyl Methoxycinnamate (and) Butyl
0.50 Methoxydibenzoylmethane (and) Ethylhexyl Salicylate (and)
PPG-26-Buteth-26 (and) PEG-40-Hydrogenated Castor oil Carbomer 0.50
Tromethamine 0.30 Demineralized water 2.00 Glycerin 3.00 Butylene
Glycol 3.00 Phenoxyethanol (and) Methylparaben (and) Butylparaben
0.70 (and) Ethylparaben (and) Propylparaben Chlorphenesin 0.25
PEG-20 Methyl Glucose Sesquistearate 3.00 Polydecene 10.00 Sweet
Almond oil 5.00 PEG-8 (and) Tocopherol (and) Ascorbyl Palmitate
(and) 0.05 Ascorbic Acid (and) Citric Acid Aldavine .TM.
(fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Polyacrylamide (and)
C13-14 Isoparaffin (and) Laureth-7 0.10 Fragrance 0.30
Example 14
TABLE-US-00012 [0212] After shave balm Ingredients Wt %
Distilled/Deionized Water QSP 100 Ethylhexyl Methoxycinnamate (and)
Butyl 0.50 Methoxydibenzoylmethane (and) Ethylhexyl Salicylate
(and) PPG-26-Buteth-26 (and) PEG-40-Hydrogenated Castor oil
Acrylates/C10-30 Alkyl Acrylate Crosspolymer 0.30 Satiaxane CX 91
0.20 Glycerin 3.00 Phenoxyethanol (and) Methylparaben (and)
Butylparaben 0.60 (and) Ethylparaben (and) Propylparaben
Chlorphenesin 0.20 Glyceryl Polymethacrylate (and) Propylene Glycol
5.00 Aluminium Starch octenylsuccinate 1.00 Squalane 2.00
Polyacrylamide (and) C13-14 isoparaffin (and) Laureth-7 0.27
Demineralized water 3.00 Tromethamine 0.30 Cyclopentasiloxane (and)
Propylene glycol 1.00 Aldavine .TM. (fucans/galactans 10/1
0.1-99.9%) 1.00-5.00 Fragrance 0.20
Example 15
TABLE-US-00013 [0213] Hair repairing mask Ingredients Wt %
Demineralized water QSP100 Propylene glycol 3.00 DMDM Hydantoin
(and) Iodopropynyl Butylcarbamate 0.10 Cetearyl Alcohol (and)
Cetearyl Glucoside 6.00 Phenyl Trimethicone 2.00 Stearyl Alcohol
3.00 Cetrimonium Chloride 1.00 Aldavine .TM. (fucans/galactans 10/1
0.1-99.9%) 1.00-5.00 Citric Acid (10% aqueous solution) 0.23
Fragrance 0.30
Example 16
TABLE-US-00014 [0214] Hand cream Ingredients Wt % Demineralized
water QSP100 Carbomer 0.15 Xanthan Gum 0.20 Sodium hydroxide (10%
aqueous solution) 0.15 Disodium EDTA 0.10 Demineralized water 2.00
Glycerin 5.00 Phenoxyethanol (and) Methylparaben (and) Butylparaben
0.80 (and) Ethylparaben (and) Propylparaben Chlorphenesin 0.28
Glyceryl Stearate (and) PEG-100 Stearate 5.00 PPG-15 stearyl Ether
2.00 Shea Butter 2.00 Cetyl Alcohol 2.50 C12-15 Alkyl Benzoate 3.00
Cyclomethicone 2.00 PEG-8 (and) Tocopherol (and) Ascorbyl Palmitate
(and) 0.08 Ascorbic acid (and) Citric acid Aldavine .TM.
(fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Polyacrylamide (and)
C13-14 isoparaffin (and) Laureth-7 0.79 Fragrance 0.20
Example 17
TABLE-US-00015 [0215] Heavy legs emulsion Ingredients Wt %
Demineralized water QSP100 Tetrasodium EDTA 0.10 Carbomer 0.30
Glycerin 4.00 Phenoxyethanol (and) Methylparaben (and) Butylparaben
0.60 (and) Ethylparaben (and) Propylparaben Chlorphenesin 0.20
Butylene Glycol 3.00 Cetearyl isononanoate 4.00 Dimethicone 3.00
Tromethamine 0.25 Demineralized water 2.00 Ethylhexyl
Methoxycinnamate (and) Butyl 0.50 Methoxydibenzoylmethane (and)
Ethylhexyl Salicylate (and) PPG-26-Buteth-26 (and)
PEG-40-Hydrogenated Castor oil Polyacrylamide (and) C13-14
isoparaffin (and) Laureth-7 2.00 Menthoxypropanediol 0.20 Aldavine
.TM. (fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Polysorbate 20
1.00 Fragrance 0.30
Example 18
TABLE-US-00016 [0216] Lips emulsion Ingredients Wt % Demineralized
water QSP100 Disodium EDTA 0.15 Xanthan Gum 0.30 Methylparaben
(and) Ethylparaben (and) Butylparaben 0.30 (and) Propylparaben
(and) isobutylparaben Chlorphenesin 0.25 Glycerin 15.00 Cetearyl
Alcohol (and) Cetearyl Glucoside 5.00 Glyceryl Stearate (and)
PEG-100 Stearate 2.00 Cetyl Alcohol 2.00 Squalane 7.00 Jojoba seed
oil 3.00 Camelina Sativa seed oil 5.00 Shea Butter 7.00 Tocopheryl
Succinate 0.15 Camelina Sativa Seed oil 1.00 Cyclomethicone 3.00
PEG-8 (and) Tocopherol (and) Ascorbyl Palmitate (and) 0.10 Ascorbic
Acid (and) Citric Acid Tocopheryl Acetate 0.25 Aldavine .TM.
(fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Fragrance 0.10 Sodium
Hydroxyde (10% aqueous solution) 0.042
Example 19
TABLE-US-00017 [0217] Slimming gel Ingredients Wt % Demineralized
water QSP100 Acrylates/C10-30 Alkyl Acrylate Crosspolymer 0.25
Tromethamine 0.15 Demineralized water 2.00 Glycerin 3.00 Benzyl
Alcohol 0.60 Phenoxyethanol 0.40 Ethylhexyl Methoxycinnamate (and)
Butyl 0.50 Methoxydibenzoylmethane (and) Ethylhexyl Salicylate
(and) PPG-26-Buteth-26 (and) PEG-40-Hydrogenated Castor oil
Aldavine .TM. (fucans/galactans 10/1 0.1-99.9%) 1.00-5.00 Fragrance
0.20 Blue 1 (0.1% aqueous solution) 0.50
Example 20
TABLE-US-00018 [0218] Cleansing cream Ingredients Wt % Glyceryl
Stearate 4.00 Cetearyl Alcohol 1.60 Sodium Cocoyl Lactylate 0.50
Mineral Oil 20.00 Mineral Oil and Lanolin Alcohol 4.00
Distilled/Deionized Water 64.70 Aldavine .TM. (fucans/galactans
10/1 0.1-99.9%) 1.00-5.00 Glycerine 5.00 Preservative 0.20
[0219] Using the FLMW/GHMW (10/1) composition in a cosmetic
formulation will potentially provide skin with the tools it needs
to protect the integrity of the microcapillaries. By inhibiting the
excessive production of VEGF and PGE.sub.2, the present composition
aims at preserving a better skin health. This phenomenon will help
protect and maintain the integrity of the extra cellular matrix
(ECM) and prevent or alleviate skin disorders as exemplified but
not limited to: those caused by UV and pollutants exposure,
sensitive skin, stress, psoriasis, acne, rosacea, telangiectasia,
skin cancer, and skin aging.
[0220] The compositions may be improved by using polysaccharides
having at least a portion of mineralized polysaccharides, namely
FHMW-M, which has for effect to increase the inhibition of
PGE.sub.2.
[0221] The compositions may be administered topically in a suitable
therapeutic formulation to animals and humans as a bioactive agent.
A dosage regimen ranging from 0.1 to 25% of 5.5 mg/ml for instance,
preferably from 1 to 10%, more preferably from 1 to 5% is
envisaged, achieving an effective dose at the site of inflammation
(local or in situ effect).
[0222] Additionally, the bioactive agent may be complexed with a
variety of well established compounds or compositions which enhance
stability or pharmacological properties such as half-life. It is
evident in the art that the therapeutic, bioactive composition may
be delivered by topical application or any other effective means or
routes which could be used for treating problems involving:
[0223] Psoriasis:
[0224] Since psoriasis appears to be a multifactorial disease, it
is assumed that the response of the patients depends on the
importance of the involvement of components like angiogenesis and
inflammation in the establishment and in the perpetuation of this
condition. Moreover excess VEGF in skin may provide just such a
predisposition by inducing a vascular inflammatory response that
then predisposes to more widespread tissue inflammation closely
resembling the psoriatic state. The ability of VEGF to induce a
psoriasiform phenotype suggests a new etiology and treatment
approach for this disease that justify the used of the present
composition in such disorder (Yu-Ping Xia, Baosheng Li, Donna
Hylton, Michael Detmar, George D. Yancopoulos, and John S. Rudge
2003 Blood, 1 Jul. 2003, Vol. 102, No. 1, pp. 161-168)
[0225] Rosacea:
[0226] A comprehensive review of this skin disorder by J. K. Wilkin
(Arch. Dermatol. (1994) vol. 130, 359-362) indicates that rosacea
develops as a combination of one or more of the following cutaneous
stigmata: flushing, erythema, telangiectesia, facial edema,
papules, pustules, ocular lesions and rhinophyma, depending on the
disorder stage development.
[0227] Hence, erythema and telangiectesia are vascular disorders,
and other characteristics of these disorders that are not of a
vascular nature, may derive from a vascular disturbance. The
erythema is the first symptom of rosacea to be observed and is
defined by an increased number of erythrocytes in a mildly inflamed
vasculature. Furthermore a dermal cellulitis may appear as a result
of an extravascular fluid accumulation consequent to irritant
factors. The edema is the result of an increased extravasation
along with a decreased fluid removal by lymphatic vessels.
Decreased lymphatic activity appears to be consequent to lymphatic
damage occurring during cellulitis. Rhinophyma may be explained by
the observation that chronic cutaneous edema is frequently followed
by connective tissue hypertrophy and fibroplasia and may also be
due to factor XIII expression. It has been further emphasized that
the elastin network that surrounds the lymphatic system in the skin
serves two important functions. First, it is a tethering that
permits the lymphatic endothelium to be sensitive to the volume of
fluids in the vicinity of the lymphatic vessels, so that any
increase of volume results in greater tension on the anchoring
filaments. Second, the elastin network provides a low-resistance
pathway through the intersticium along which micromolecules pass to
the lymphatic vessels. Elastin degeneration due to actinic exposure
is probably a common cause of lymphatic failure in rosacea.
[0228] During inflammation, neutrophils are recruited and
exacerbate the rapid degradation of a variety of extracellular
matrix macromolecules, especially elastin. Neutrophil elastase
degrades type IV collagen in the extracellular matrix on which the
integrity of the capillary walls depends. When lymphatic failure
occurs, a sustained inflammation takes place. When lymphatic
failure does not resolve the inflammation becomes self-sustained.
The plasma proteins that accumulate in the sustained inflamed
tissue appear to contribute to the fibroplasia, which underlies the
development of rhinophyma.
[0229] Telangiectesia represents the latter phase of the vascular
stage of rosacea. The mechanical integrity of the dermal connective
tissue is reduced, allowing a passive dilation of the vasculature.
The perivascular inflammatory cells thus infiltrate and contribute
to rosacea. Dilatation of both dermal blood vessels and lymphatics
are prominent in rosacea. Angiogenesis may contribute to the
telangiectesia. Angiogenesis depends on the space left between in a
tissue where endothelial cells can grow. Edema reduces the tissue
compactness, permitting vascularization. Since lymphatic failure
results in sustained inflammatory response, the edema thus created
would be one the feature that favors angiogenesis.
[0230] As already mentioned above, it appears that rosacea is
initiated by an inflammatory reaction which does not resorb with
time. Since the polysaccharide compositions of the present
invention comprise a plurality of biological activities such as an
anti-inflammatory activity and since it inhibits VEGF secretion, it
is contemplated that the polysaccharide compositions of the present
invention will have an effect on all skin diseases involving one or
more etiologies related to these biological activities. The
compositions of the present invention comprise a plurality of
active ingredients and as such will be effective in treating mono-
as well as in pluri-factorial diseases or disorders. The treatment
of rosacea is a typical example of such a plurifactorial disorder
wherein inflammation, and extravasation occur.
[0231] Other or similar compositions can also be conceived to be
used in a method for soothing skin or for reducing inflammation in
mammalian skin. Inflammation can be caused by various agents such
as chemical irritant, physical abrasion and exposure to ultraviolet
radiation. Compositions and methods for inhibiting collagenase in
skin are also contemplated. Collagenase and inflammation are linked
to premature aging (degradation of collagen), and therefore the
antagonist activities recovered in the polysaccharide compositions
could also be put to contribution in compositions and methods for
retarding premature aging, and for regulating wrinkles or atrophy
in mammalian skin. As causes of wrinkles or atrophy are listed, by
way of examples, age, exposure to ultraviolet radiation or to
environmental pollutant. Topical compositions may comprise an
effective amount of polysaccharide composition, to be determined
for each specific application.
[0232] Acne: In acne pathogenesis, Propionibacterium acnes is known
to induce inflammatory mediators, such as IL-8. This cytokine may
be directly implicated in the inflammatory response by its capacity
to attract, by a chemotactic response, inflammatory cells to the
site of infection. IL-8 can also be implicated in the development
of acne lesions (Schaller M, Loewenstein M, Borelli C, Jacob K,
Vogeser M, Burgdorf W H C, Plewig G. Induction of a chemoattractive
proinflammatory cytokine response after stimulation of
keratinocytes with Propionibacterium acnes and coproporphyrin III.
Br J. Dermatol. 2005 July; 153(1):66-71.). According to this
pathogenesis, topical compositions may be conceived to be used to
reduce inflammatory response and acne lesions.
[0233] In general, these compositions may contain from about 0.1 to
about 30 weight percent of a polysaccharide compositions and from
about 70 to 99.9 weight percent of a pharmaceutically acceptable
vehicle. These compositions may contain an anti-oxidant, such as an
agent which prevents the formation of lipid peroxides in skin.
Examples of such an anti-oxidant are tocopherol, tocopherol
derivatives, ascorbic acid, ascorbic acid derivatives and BHT. The
compositions can be complemented with anti-inflammatory agents like
a phospholipase A2 inhibitor or the botanically-derived
anti-irritants cola, green tea and cartilage extracts. Topical
compositions may take diverse forms such as solutions, suspensions,
lotions, tinctures, gels, creams, sprays, emulsions, sticks,
ointments or liposomes. Other cosmetic applications include dark
circle around the eyes and skin barrier function, care of skin
photo-damaging, sensitive skin, erythema, rosacea, teleangiectasia,
actinic elastosis, psoriasis, and acne.
[0234] Although the present invention has been described
hereinabove by way of specific embodiments thereof, it can be
modified, without departing from the spirit and nature of the
subject invention as defined in the appended claims.
* * * * *