Topical compositions comprising fucan and galactan and their use for decreasing skin thickness

Abstract

Stable topical composition comprising fucan and galactan in a dry weight ratio of between about 0.75 : about 1 (w / w) and about 20 : about 1 (w / w), water, a first preservative agent comprising at least 3% w / w glycerin, and a second preservative agent comprising phenylpropanol, and uses thereof to decrease epidermis thickness and / or skin cell proliferation in a subject in need thereof.

Description

[0001] TITLE

[0002] TOPICAL COMPOSITIONS COMPRISING FUCAN AND GALACTAN AND THEIR USE FOR DECREASING SKIN THICKNESS

[0003] FIELD OF THE DISCLOSURE

[0004] The present disclosure relates to topical compositions comprising fucan and galactan and their use for decreasing skin thickness. More specifically, the present disclosure is concerned with topical compositions comprising fucan and galactan comprising certain preservative agents and their use for decreasing skin thickness and / or skin cell proliferation.

[0005] BACKGROUND OF THE DISCLOSURE

[0006] Although the term “sensitive skin” does not have a clear definition in the art, it is generally used to refer to skin presenting certain characteristics and symptoms such as a reduced skin barrier function, an increased transcutaneous penetration, which is characterized by a reduced loricrin expression in skin cells, increased skin proliferation, increased skin thickness, and an hyperreactivity to external or internal triggering factors. Triggering factors can be environmental conditions such as wind, pollution, temperature variations, excessively hard water, low hygiene, cosmetic or care products, diets or certain medicaments. Triggering factors can also be associated with stress in the subject. Subjects with sensitive skin may experience a burning sensation and / or tingling sensation in their skin.

[0007] There is a need for compositions treating at least certain symptoms of sensitive skins.

[0008] The present description refers to a number of documents, the content of which is herein incorporated by reference in their entirety.

[0009] SUMMARY OF THE DISCLOSURE

[0010] More specifically, in accordance with the present disclosure, there are provided the following items:

[0011] Item 1 . Use of a topical composition comprising fucan and galactan in a dry weight ratio of between about 0.75 : about 1 (w / w) and about 20 : about 1 (w / w), water, a preservative agent comprising at least 3% w / w glycerin, and a further preservative agent comprising at least one of (i) glyceryl caprylate; (ii) aqua (and) glycerin (and) sodium levulinate; (iii) a further amount of at least 15% w / w glycerin; and (iv) sodium benzoate, to decrease epidermis thickness and / or skin cell proliferation in a subject in need thereof.

[0012] Item 2. The use of item 1 , to (i) decrease at least one of ki67, keratin 14 (K14), cytoplasmic Linker Associated Protein 2 (Clasp2) and Wnt family member 5A (Wnt5a) expression; and / or to (ii) increase loricrin expression in skin cells in a subject in need thereof.

[0013] Item 3. The use of item 2, wherein at least two of ki67, K14, Clasp2 and Wnt5a expression in skin cells are decreased.

[0014] Item 4. The use of any one of items 1-3, wherein the composition comprises fucan : galactan in a dry weight ratio of between about 12 : about 1 (w / w) and about 18: about 1 (w / w). Item 5. The use of any one of items 1-3, wherein the fucan is extracted from brown algae and / or galactan is extracted from red algae.

[0015] Item 6. The use of any one of items 1-3, wherein the fucan is extracted from Ascophyllum nodosum and / or the galactan is extracted from Asparagopsis armata.

[0016] Item 7. The use of any one of items 1-6, wherein the further preservative agent comprises at least one of glyceryl caprylate , aqua (and) glycerin (and) sodium levulinate; and sodium benzoate.

[0017] Item 8. The use of any one of items 1-7, wherein the topical composition further comprises sorbitol.

[0018] Item 9. The use of any one of items 1 -8, wherein the topical composition further comprises potassium sorbate.

[0019] Item 10. A topical composition comprising fucan and galactan in a dry weight ratio of between about 0.75 : about 1 (w / w) and about 20 : about 1 (w / w), water, a preservative agent comprising at least 3% w / w glycerin, and a further preservative agent that comprises at least one of (i) glyceryl caprylate; (ii) aqua (and) glycerin (and) sodium levulinate; (iii) a further amount of at least 15% w / w glycerin; and (iv) sodium benzoate.

[0020] Item 11. The composition of item 10, comprises fucan : galactan in a dry weight ratio of between about 12 : about 1 (w / w) and about 18: about 1 (w / w).

[0021] Item 12. The composition of item 10 or 11, wherein the fucan is extracted from brown algae and / or galactan is extracted from red algae.

[0022] Item 13. The composition of any one of items 10-12, wherein the fucan is extracted from Ascophyllum nodosum and / or the galactan is extracted from Asparagopsis armata.

[0023] Item 14. The composition of any one of items 10-13, wherein the further preservative agent comprises at least one of glyceryl caprylate, aqua (and) glycerin (and) sodium levulinate; and sodium benzoate.

[0024] Item 15. The composition of any one of items 10-14, further comprising sorbitol and / or potassium sorbate.

[0025] Item' 1. Use of a topical composition comprising fucan and galactan in a dry weight ratio of between about 0.75 : about 1 (w / w) and about 20 : about 1 (w / w), water, a first preservative agent comprising at least 3% w / w glycerin, and a second preservative agent comprising phenylpropanol, to decrease epidermis thickness and / or skin cell proliferation, in a subject in need thereof.

[0026] Item' 2. The use of item’ 1 , wherein the composition comprises fucan : galactan in a dry weight ratio of between about 12 : about 1 (w / w) and about 18: about 1 (w / w).

[0027] Item' 3. The use of item’ 1 or 2, wherein the fucan is extracted from brown algae and / or galactan is extracted from red algae.

[0028] Item' 4. The use of any one of items’ 1-3, wherein the fucan is extracted from Ascophyllum nodosum and / or the galactan is extracted from Asparagopsis armata.

[0029] Item' 5. The use of any one of items. 1-4, wherein the second preservative agent consists of phenylpropanol. Item' 6. The use of any one of items' 1-5, wherein the second preservative consists of about 0.2 to 0.4 % w / w phenylpropanol.

[0030] Item' 7. The use of any one of items’ 1 -6, wherein the first preservative agent comprises or consists of between about 14% w / w and about 65% w / w of glycerin.

[0031] Item' 8. The use of any one of items’ 1 -7 , wherein the first preservative agent comprises or consists of between about 20% w / w and about 60% w / w of glycerin, or about 30% w / w and about 60% w / w of glycerin, or about 40% w / w and about 60% w / w of glycerin, or about 45% w / w and about 55% w / w of glycerin or about 50% w / w and about 53% w / w of glycerin.

[0032] Item' 9. A stable topical composition comprising fucan and galactan in a dry weight ratio of between about 0.75 : about 1 (w / w) and about 20 : about 1 (w / w), water, a first preservative agent comprising between about 14% w / w and about 65% w / w glycerin, and a second preservative agent that comprises between about 0.2 to 0.4 % w / w phenylpropanol.

[0033] Item' 10. The composition of item’ 9, comprises fucan : galactan in a dry weight ratio of between about 12 : about 1 (w / w) and about 18: about 1 (w / w).

[0034] Item' 11 . The composition of item’ 9 or 10, wherein the fucan is extracted from brown algae and / or galactan is extracted from red algae.

[0035] Item' 12. The composition of any one of items’ 9-11, wherein the fucan is extracted from Ascophyllum nodosum and / or the galactan is extracted from Asparagopsis armata.

[0036] Item' 13. The composition of any one of items’ 9-12, wherein the second preservative agent consists of phenylpropanol.

[0037] Item' 14. The composition of any one of items’ 9-12, wherein the second preservative consists of about 0.2 to 0.4 % w / w phenylpropanol.

[0038] Item' 15. The composition of any one of items’ 9-14, wherein the first preservative agent comprises or consists of between about 14% w / w and about 65% w / w of glycerin.

[0039] Item' 16. The composition of item’ 15, wherein the first preservative agent comprises or consists between about 20% w / w and about 60% w / w of glycerin, or about 30% w / w and about 60% w / w of glycerin, or about 40% w / w and about 60% w / w of glycerin, or about 45% w / w and about 55% w / w of glycerin or about 50% w / w and about 53% w / w of glycerin.

[0040] Item' 17. The use of any one of items’ 1-8 or the composition of any one of clams 9-16, wherein the galactan is in the form of a syrup comprising galactan, glycerin, and phenylpropanol.

[0041] Item' 18. The use or composition of item’ 17, wherein the syrup comprises about 50% w / w (e.g., 50.225 % w / w) of galactan, about 49% w / w (e.g., 49.475 % w / w) of glycerin and about 0.3% w / w phenylpropanol.

[0042] Item' 19. The use or the composition of item’ 18, wherein the composition comprises about 6.25 % w / w of galactan syrup and about 2.5 % w / w fucan. Item' 20. ’Stable topical composition comprising:

[0043] Item' 21 . about 40.95% w / w water;

[0044] Item' 22. about 6.25 % w / w of galactan syrup;

[0045] Item' 23. about 2.5 % w / w of fucan;

[0046] Item' 24. about 50% w / w glycerin; and

[0047] Item' 25. about 0.3% w / w phenylpropanol.

[0048] Item' 26. ’ The composition of item’ 20, wherein the galactan syrup comprises galactan, glycerin, and phenylpropanol.

[0049] Item' 27. The composition of item’ 20 or 21, wherein the galactan syrup comprises about 50% w / w (e.g., 50.225 % w / w) of galactan, about 49% w / w (e.g., 49.475 % w / w) of glycerin and about 0.3% w / w phenylpropanol.

[0050] Item' 28. 'Use of the composition defined in any one of items’ 20-22, to decrease epidermis thickness and / or skin cell proliferation in a subject in need thereof.

[0051] In specific embodiments, the subject in need thereof is a human.

[0052] Other objects, advantages and features of the present disclosure 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.

[0053] BRIEF DESCRIPTION OF THE DRAWINGS

[0054] In the appended drawings:

[0055] [FIG. 1]: Effect of Aldavine™ 5X treatment on the epidermis thickness. Skin substitutes were untreated (black bar) or treated with Aldavine™ 5X at 1 % or 2% (white bars) or with Methotrexate (MTX, grey bar) at 734 pM used as positive control. The data represents the quantification of epidermal thickness normalized to 100% of untreated condition.

[0056] [FIG. 2]: Effect of Aldavine™ 5X treatment on the Ki67 expression. Skin substitutes were untreated (black bar) or treated with Aldavine™ 5X at 1 % or 2% (white bars) or with Methotrexate (MTX, grey bar) at 734 pM used as positive control. The data represents the quantification of KI67 positive cells normalized to 100% of untreated condition.

[0057] [FIG. 3]: Effect of Aldavine™ 5X treatment on the K14 expression. Skin substitutes were untreated (black bar) or treated with Aldavine™ 5X at 1% or 2% (white bars) or with Methotrexate (MTX, grey bar) at 734 pM used as positive control. The data represents the quantification of K14 intensity related to the area and normalized to 100% of untreated condition.

[0058] [FIG. 4]: Effect of Aldavine™ 5X treatment on CLASP2 expression. Skin substitutes were untreated (black bar) or treated with Aldavine™ 5X at 1 % or 2% (white bars) or with Methotrexate (MTX, grey bar) at 734 pM used as positive control. The graph presented the quantification of Clasp2 expression related to total proteins and normalized to 100% of untreated condition.

[0059] [FIG. 5]: Effect of Aldavine™ 5X treatment on Wnt5a expression. Skin substitutes were untreated (black bar) or treated with Aldavine™ 5X at 1% or 2% (white bars) or with Methotrexate (MTX, grey bar) at 734 pM used as positive control. The data represents the quantification of Wnt5a intensity related to the area and normalized to 100% of untreated condition.

[0060] [FIG. 6]: Effect of Aldavine™ 5X treatment on Loricrin expression. Skin substitutes were untreated (black bar) or treated with Aldavine™ 5X at 1% or 2% (white bars) or with Methotrexate (MTX, grey bar) at 734 pM used as positive control. The data represents the quantification of Loricrin intensity related to the cell number and normalized to 100% of untreated condition.

[0061] DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0062] The present disclosure provides uses of a topical composition comprising fucan and galactan and a preservative agent for decreasing at least one of skin cell proliferation, skin thickness, ki67 expression in skin cells, K14 expression in skin cells, Clasp2 expression in skin cells and Wnt5a expression in skin cells; and / or increasing loricrin expression in skin cells in a subject in need thereof. The present disclosure relates to a use of the topical composition for treating, preventing or alleviating the symptoms of disorders and diseases associated with excessive skin cell proliferation and / or thickness.

[0063] As used herein skin cells refers to at least one of fibroblasts and keratinocytes.

[0064] 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. Fucans obtained 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 a(1 — >3) and a(1 — >4) glycosidic bonds.

[0065] 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.

[0066] 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 1]: Red algae as sources of galactans and known processes for their obtention

[0067] [Table 2]: Red algae as sources of galactans and known processes for their obtention

[0068] As used herein the term “fucans” refers to polysaccharides comprising sulfated fucose and uronic acid. Without being so limited, Laminariales, Chordariales, Fucales marine algae including Kjellmaniella crassifolia, Laminaria japonica, Kjellmaniella, Fucus, Nemacystus, Cladosiphon okamuranus, Undaria, Undaria pinnatifida (Wakame Mekabu), Ecklonia kurome, Stichopus japonicus, Eisenia, Ecklonia, Giant kelp, Lessonia nigrescence, Gelidiun amansii, Gracilaria, Pteroclavia capillacae, Sargassum sp., Padina gymnospora, Adenocystis utricularis, Pelvetia canaliculata, Chorda filum and Ascophyllum nodosum along with those derived from Echinodermata, sea cucumber, Echnoidea and Asterozoa contain fucans suitable for uses and methods of the present disclosure.. Fucans of the present disclosure are exemplified herein by fucans derived from the brown algae Ascophyllum nodosum.

[0069] 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, Cryptopleura ramosa, Chondrus ascellatus, Gymnogongrus tolurus, Phacelocarpus peperocarpos, and Sargassum kjellmanianum, contain galactans suitable for uses and methods of the present disclosure. The galactans of the present disclosure are exemplified herein by galactans derived from the red algae Asparagopsis armata.

[0070] Preparation of Fucans

[0071] Fucans from various algae may be prepared by known methods and the resulting purified products, fucans-containing products can be used in the present disclosure.

[0072] In a specific embodiment, fucans of the present disclosure 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 a(1— >3) and a(1 — >4) glycosidic bonds.

[0073] Native Fucans (i.e., which contain mil

[0074] 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.

[0075] The algae could be chopped with instruments including but not limited to, a meat chopper, kitchen homogenizer, Polytron™ 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.

[0076] 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 disclosure. 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.

[0077] 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.

[0078] The supernatant is then separated from the pellet (solid / liquid separation). 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.

[0079] 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.

[0080] The clarified material is then cooled down at a temperature ranging from about 15 to about 30 degrees Celsius, preferably from about 15 to about 25 degrees Celsius.

[0081] 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.

[0082] 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.

[0083] 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 fucan fraction which is naturally mineralized.

[0084] 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 (SO4) radical.

[0085] According to another embodiment, fucans could be extracted from the chopped algae by an adequate volume of waterbased 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 filtration ultrafiltration.

[0086] Depolymerized Fucans

[0087] 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 fraction is 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.

[0088] The solution is then neutralized with NaOh and then concentrated / purified by ultrafiltration followed by a dialysis.

[0089] 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.

[0090] Demineralised Fucans

[0091] 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+, CI-) 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. of Galactans

[0092] In a specific embodiment, galactans of the present disclosure 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.

[0093] 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.

[0094] Native Galactans (i.e. , which contain mil

[0095] 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 disclosure. 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.

[0096] 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.

[0097] The supernatant is then separated from the pellet (solid / liquid separation). 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.

[0098] 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.

[0099] 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.

[0100] 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.

[0101] 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.

[0102] 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.

[0103] 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.

[0104] 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.

[0105] 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 (SO4) radical.

[0106] Estimation of the molecular weight of polysaccharides formed by HPSEC (High Performance Size Exclusion

[0107] 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 oligosaccharide, 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 glucans, 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™ G3000 SW-XL and TSK™ 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 mL.min-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.

[0108] In accordance with a specific embodiment of the present disclosure, 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 100kDa and in more specific embodiments, the Mw was about 350 kDa (obtained from a pullulan equivalent).

[0109] Galactans

[0110] 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.

[0111] The solution is then neutralized and then concentrated / purified on a 2kDa ultrafiltration followed by a dialysis.

[0112] 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 galactan fraction.

[0113] Demineralized Galactans

[0114] The galactan 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.

[0115] Fucan: galactan

[0116] According to still other specific embodiments, the galactans have an average molecular weight higher than about 100kDa. According to still other specific embodiments, the galactans have an average molecular weight of about 350kDa. 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 10OkDa. According to still other specific embodiments, the fucans have an average molecular weight ranging from about 5kDa to about 25kDa. 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 dry weight ratio fucans: galactans present in the composition is between about 2.5 : about 1 (w / w) to about 40 : about 1 (w / w), or about 0.75 : about 1 (w / w) to about 20: about 1 (w / w), or between about 1 : about 1 (w / w) to about 20: about 1 (w / w), or between about 1 : about 1 (w / w) to about 19: about 1 (w / w), or between about 1 : about 1 (w / w) to about 18: about 1 (w / w), or between about 1 : about 1 (w / w) to about 17: about 1 (w / w), or between about 1 : about 1 (w / w) to about 16: about 1 (w / w), or between about 1 : about 1 (w / w) to about 15: about 1 (w / w), or between about 1 : about 1 (w / w) to about 14: about 1 (w / w), or between about 1 : about 1 (w / w) to about 13.5: about 1 (w / w), or between about 2 : about 1 (w / w) to about 20: about 1 (w / w), or between about 3 : about 1 (w / w) to about 19: about 1 (w / w), or between about 4 : about 1 (w / w) to about 18: about 1 (w / w), or between about 5 : about 1 (w / w) to about 17: about 1 (w / w), or between about 6 : about 1 (w / w) to about 16: about 1 (w / w), or between about 7 : about 1 (w / w) to about 15: about 1 (w / w), or between about 8 : about 1 (w / w) to about 14: about 1 (w / w), or between about 9 : about 1 (w / w) to about 13.5: about 1 (w / w), or about 13.16: about 1 (w / w). According to still other specific embodiments, the fucans and the galactans are present a ratio of about 10 fucans: about 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.

[0117] Active ii

[0118] Without being so limited, the compositions of the present disclosure may further comprise at least one further active ingredient such as an anesthetic 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, antiproliferation agent, antipruritic agent, anti-rosacea agent, anti-seborrhea agent, anti-stress agent, antitelangiectasia 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, makeup 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, vitamin D, vitamin D analogue, corticosteroid, retinoid, calcineurin inhibitor, salicylic acid, coal tar, anthralin, methotrexate, ceramides, niacinamide, Abyssine™, Borealine™ Protect, Canadian Willowherb™, Neutrazen™, Tazman pepper, 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 fucans and galactans of the present disclosure. In a specific embodiment, the further active ingredient is an antiproliferation agent, vitamin D, vitamin D analogue, corticosteroid, retinoid, calcineurin inhibitor, salicylic acid, coal tar, or anthralin.

[0119] 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, |3-carotene, retinyl ester), vitamin D and its derivatives (cholecalciferol, ergocalciferol, 25- hydroxycholecalciferol), growth factors and estradiol derivatives.

[0120] Without being so limited, anaesthetics 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.

[0121] 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, |3-carotene, retinyl ester), salicylic acid, sulfur, sulfurated lime, alcohol and acetone.

[0122] 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, |3-carotene, retinyl ester), epidermal growth factor, ceramide, ethylbisiminomethylguaiacol manganese chloride, glycation inhibitors, chrysanthemum indicum extract and aphanizomenon flos aquae extract.

[0123] 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.

[0124] 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.

[0125] 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 (a-tocopherol, 5-tocopherol, y-tocopherol), chamomile oil, gingko biloba oil and camellia sinensis extract.

[0126] 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.

[0127] 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 (a-tocopherol, 5-tocopherol, y- tocopherol), grape seed extract and camellia sinensis extract.

[0128] Without being so limited, antiproliferation agents include mitomycine C and methotrexate.

[0129] 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, and cyproheptadine.

[0130] 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.

[0131] 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.

[0132] 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.

[0133] 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.

[0134] 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, |3-carotene, retinyl ester), vitamin C and its derivatives (ascorbyl palmitate, magnesium ascorbyl phosphate, sodium ascorbyl phosphate), growth factors and its derivatives.

[0135] 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 alph a / beta hydroxy acids, salicylic acid, glycolic acid, lactic acid, citrus acid and walnut shell powder.

[0136] 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.

[0137] 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™-like), chitosan, arnica extract, fennel-sweet oil and papaya extract.

[0138] Without being so limited, free radical scaven ging / anti pol I ution / 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).

[0139] 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.

[0140] 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, - carotene, retinyl ester) and hydroxamate derivatives.

[0141] 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.

[0142] 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.

[0143] 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, chrysanthemum indicum extract, camellia sinensis extract and alteromonas ferment extract.

[0144] 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 (a-tocopherol, 5-tocopherol, y- tocopherol).

[0145] 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, |3-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.

[0146] 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 chrysanthemum indicum extract, dihydromyricetin, theobromine, theophylline, aminophylline, caffeine, isopropylarterenol hydrochloride, epinephrine, a-MSH agonists, adenylate cyclase activators and phosphodiesterase inhibitors.

[0147] 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 (a-tocopherol, 5-tocopherol, y-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, |3-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.

[0148] 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, a-MSH agonists, adenylate cyclase activators and phosphodiesterase inhibitors.

[0149] 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.

[0150] 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, |3-carotene, retinyl ester), ruminex occidentalis extract, licorice, mulberry, arctostaphylos uva-ursi (bearberry), tyrosinase inhibitors, melanosome-transfer inhibitors and melanin scavengers.

[0151] The compositions of the present disclosure 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.

[0152] 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.

[0153] 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.

[0154] Without being so limited, chelating agents are ingredients capable of binding mono and divalent cations, such as tetrasodium EDTA and disodium EDTA.

[0155] Without being so limited, conditioner agents are ingredients with lubricating action and hydrating effect, such as cetrimonium chloride, dicetyldimonium chloride, trideceth-12, quaternium-Z7, quaternium-18, polyquaternium-10, behentrimonium methosulfate, cetearyl alcohol, stearamidopropyl dimethylamine, trimethylsilylamodimethicone, isolaureth-6, octoxynol-4, dimethicone, dimethiconol, cyclopentasiloxane, pareth-7, pareth-9, linoleic acid and glycerin.

[0156] 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.

[0157] 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.

[0158] 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, sorbitol, 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.

[0159] 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 trimethylolpropane siloxy silicate, trimethylsiloxysilicate, dimethicone, vinyl dimethicone and cyclopentasiloxane.

[0160] 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-ethylhexylsulfate. Without being so limited, humectant agents are ingredients capable of maintaining constant humidity and retaining moisture, such as glycerin, PEG-8, butylene glycol and propylene glycol.

[0161] Without being so limited, lubricant agents are ingredients capable of adding slipperiness and reducing friction to improve application, such as dimethicone and dimethicone copolyol.

[0162] Without being so limited, neutralizer agents are ingredients capable of changing the acid-alkaline balance, such as triethanolamine and sodium hydroxide.

[0163] 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.

[0164] Without being so limited, preservative agents are ingredients capable of retarding or preventing microbial or chemical spoilage and protecting against discoloration, such as at least one of glycerin; phenylpropanol (e.g., A-Leen™ Aroma- 3), glyceryl caprylate (e.g., Dermosoft™ GMCY MB or Softisan™ GC8 Palm free); caprylhydroxamic acid (and) caprylyl glycol (and) glycerin (e.g.., Spectrastat™ G2 Natural MB), aqua (and) glycerin (and) sodium levulinate (and) Sodium Anisate (e.g., Dermosoft™ 1388 ECO NAL); potassium sorbate; and sodium benzoate; or any combination thereof. In specific embodiments, the preservative agent is glycerin in combination with at least one of phenylpropanol and glyceryl caprylate (Dermosoft™ GMCY MB or Softisan™ GC8 Palm free). In specific embodiments, the preservative agent is glycerin in a concentration of about 3% to about 90% w / w, or 3% to 15%, or 3% to 20%, or 3% to 25%, or 3% to 30%, or 3% to 35%, or 3% to 40%, or 3% to 45%, or 3% to 50%, or 3% to 55%, or 3% to 60%, or 3% to 65%, or 3% to 70%, or 3% to 75%, or 3% to 85%, or about 3%, or about 18%, or about 73% or about 74%. In specific embodiments, the preservative agent is glycerin in a concentration of about 3% to about 90% and one or more further preservative agents selected from glyceryl caprylate (e.g., Dermosoft™ GMCY MB or Softisan™ GC8 Palm Free), aqua (and) glycerin (and) sodium levulinate (and) Sodium Anisate (e.g., Dermosoft™ 1388 ECO NAL), potassium sorbate and sodium benzoate, each in a concentration of between about 0.01 % to about 10%, or between about 0.1% to about 10%, or between about 0.05% to about 10%, or between about 0.06% to about 10%, or between about 0.07% to about 10%, or between about 0.08% to about 10%, or between about 0.09% to about 10%, or between about 0.1 % to about 10%, or between about 0.1% to about 8%, or between about 0.1% to about 7%, or between about 0.1 % to about 6%, or between about 0.1 % to about 5%, or between about 0.1 % to about 4%, or between about 0.1% to about 3%, or between about 0.1% to about 2%, or between about 0.1% to about 1 %, or between about 0.1% to about 0.9%, or between about 0.1% to about 0.8%, or between about 0.1 % to about 0.7%, or between about 0.1% to about 0.6%, or between about 0.1% to about 0.5%, or between about 0.1 % to about 0.4%, or between about 0.1% to about 0.3%, or between about 0.1% to about 0.2%, or between about 0.2% to about 10%, or between about 0.2% to about 8%, or between about 0.2% to about 7%, or between about 0.2% to about 6%, or between about 0.2% to about 5%, or between about 0.2% to about 4%, or between about 0.2% to about 3%, or between about 0.2% to about 2%, or between about 0.2% to about 1%, or between about 0.2% to about 0.9%, or between about 0.2% to about 0.8%, or between about 0.2% to about 0.7%, or between about 0.2% to about 0.6%, or between about 0.2% to about 0.5%, or between about 0.2% to about 0.4%, or between about 0.2% to about 0.3%, or between about 0.3% to about 10%, or between about 0.3% to about 8%, or between about 0.3% to about 7%, or between about 0.3% to about 6%, or between about 0.3% to about 5%, or between about 0.3% to about 4%, or between about 0.3% to about 3%, or between about 0.3% to about 2%, or between about 0.3% to about 1%, or between about 0.3% to about 0.9%, or between about 0.3% to about 0.8%, or between about 0.3% to about 0.7%, or between about 0.3% to about 0.6%, or between about 0.3% to about 0.5%, or between about 0.3% to about 0.4%, or between about 0.4% to about 10%, or between about 0.1% to about 5%, or between about 0.1% to about 4%, or between about 0.1% to about 3%, or between about 0.1 % to about 2.5%.

[0165] Without being so limited, solubilizer agents are ingredients capable of allowing incompatible ingredients to become part of a homogeneous solution, such as sorbitol, polysorbate, ceteareth, steareth and PEG.

[0166] 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.

[0167] 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.

[0168] 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.

[0169] 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.

[0170] 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.

[0171] 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,

[0172] The composition of the present disclosure 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, and unit dosages.

[0173] The compositions of the present disclosure 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.

[0174] Applications of the compositions of the present disclosure 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.

[0175] As used herein the terms “subject” or “subject in need thereof” refer to a subject who would benefit from receiving an effective amount of the composition of the present disclosure. It refers to an animal, a mammal and to a human in a specific embodiment. In specific embodiments, the subject has a sensitive skin. The compositions of the present disclosure may also be used for preclinical uses in other animals.

[0176] The use of the terms "a" and "an" and "the" and similar referents in the context of describing the disclosure (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0177] The terms "comprising", "having", "including", and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to") unless otherwise noted.

[0178] Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All subsets of values within the ranges are also incorporated into the specification as if they were individually recited herein.

[0179] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context.

[0180] The term "about" has its ordinary meaning. In embodiments, it may mean plus or minus 10% of the numerical value qualified. Herein, the term "approximately" has its ordinary meaning. In embodiments, it may mean plus or minus 10% of the numerical value qualified.

[0181] The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the disclosure and does not pose a limitation on the scope of the disclosure unless otherwise claimed.

[0182] No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0183] The present disclosure is illustrated in further details by the following non-limiting examples. EXAMPLE 1 : Material and methods of fucoidan used in Aldavine™ 5X 1% and 2%

[0184] • Pretreatment in formaldehyde (Polyphenol trap);

[0185] • Aqueous extraction;

[0186] • Solid / liquid separation;

[0187] • Micro-particles removal;

[0188] • Alginates precipitation and removal;

[0189] • Filtration;

[0190] • Depolymerization by chemical hydrolysis; and

[0191] • Dehydration and homogenization of powder granulometry (atomization).

[0192] The resulting fucoidan has a medium molecular weight (M MW) between 20,000 to 50,000 Da and is designated herein “fucan powder'1. used in Aldavine™ 5X 1% and 2%

[0193] • Aqueous extraction;

[0194] • Solid / liquid separation;

[0195] • Micro-particles removal;

[0196] • Filtration;

[0197] • Demineralization to produce an aqueous galactan extract;

[0198] • Formulation of galactan syrup.

[0199] The composition of the galatan syrup used in each of formulations A1 to A19 is presented in Table 3 below.

[0200] [Table 3]: Galactan syrup formulations

[0201] Aldavine™ 5X

[0202] Manufacturing process of Aldavine™ 5X

[0203] Compositions / formulations comprising fucans powder: galactans sirup at a ratio of 1 : 2.5, water, and one or more preservative agents, and eventually sorbitol were prepared as follows. The 1% w / w Aldavine™ 5X formulation contains 0.025% w / w fucans powder and 0.0625 % w / w galactans syrup (3% Solid matter i. e. 1 .9 mg / g solid matter) in culture medium and the 2% w / w Aldavine™ 5X formulation contains 0.05% w / w fucans powder and 0.125 % w / w galactans syrup. These compositions were used in assays presented in Examples 2 to 10.

[0204] The composition of each of formulations A1 to A19 is presented in Table 4 below.

[0205] [Table 4]: Aldavine™ 5X formulations

[0206] a: galactan syrup G2; b: galactan syrup G3; c: galactan syrup G4; d: ga actan syrup G1 ; e: galactan syrup G5; f: galactan syrup G6; g: galactan syrup G7; h: galactan syrup G8; anc i: galactan syrup G9.

[0207] Process Aldavine™ 5X - A1 formulation

[0208] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0209] Add Dermosoft™ GMCY MB and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0210] Add galactan syrup G2 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0211] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0212] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0213] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0214] Filter the product through a 40-mesh or 60-mesh sieve.

[0215] Process Aldavine™ 5X - A2 formulation

[0216] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0217] Add Spectrastat™ G2 Natural MB and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0218] Add galactan syrup G3 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0219] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0220] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0221] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0222] Filter the product through a 40-mesh or 60-mesh sieve.

[0223] Process Aldavine™ 5X - A3 formulation

[0224] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0225] Add Spectrastat™ G2 Natural MB and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0226] Add galactan syrup G4 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0227] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0228] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C. Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0229] Filter the product through a 40-mesh or 60-mesh sieve.

[0230] Process Aldavine™ 5X - A4 formulation

[0231] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0232] Add the glycerin. Shake for 5 minutes at 60+ / -3°C.

[0233] Add galactan syrup G1 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0234] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0235] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0236] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0237] Filter the product through a 40-mesh or 60-mesh sieve.

[0238] Process Aldavine™ 5X - A5, A7 and A8 formulation

[0239] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0240] Add the glycerin. Shake for 5 minutes at 60+ / -3°C. Add galactan syrup (G1 for A5, G2 for A7, G4 for A8 (see Table 3)). Shake for 5 minutes at 60+ / -3°C.

[0241] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0242] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0243] Filter the product through a 40-mesh or 60-mesh sieve.

[0244] Process Aldavine™ 5X - A6 formulation

[0245] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0246] Add phenoxyethanol. Shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0247] Add potassium sorbate. Shake for 5 minutes at 60+ / -3°C.

[0248] Add galactan syrup G5 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0249] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C. Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0250] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0251] Filter the product through a 40-mesh or 60-mesh sieve.

[0252] Process Aldavine™ 5X - A9 formulation

[0253] Shake the glycerin. Heat to 60+ / -3°C.

[0254] Add Dermosoft™ GMCY MB and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0255] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 30 minutes or until the solution is homogeneous at 60+ / -3°C.

[0256] Add purified water. Shake for 5 minutes or until solution is homogeneous at 60+ / -3°C.

[0257] Add galactan syrup G2 (see Table 3). Shake for 5 minutes or until solution is homogeneous at 60+ / -3°C.

[0258] With rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0259] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0260] Filter the product through a 40-mesh or 60-mesh sieve.

[0261] Process Aldavine™ 5X - A10 formulation

[0262] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0263] Add potassium sorbate. Shake for 5 minutes at 60+ / -3°C.

[0264] Add sodium benzoate. Shake for 5 minutes at 60+ / -3°C.

[0265] Add galactan syrup G6 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0266] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0267] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0268] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0269] Filter the product through a 40-mesh or 60-mesh sieve.

[0270] Process Aldavine™ 5X - A11 formulation Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0271] Add Dermosoft™ 1388 ECO NAL. Shake for 5 minutes at 60+ / -3°C.

[0272] Add galactan syrup G7 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0273] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0274] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0275] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0276] Filter the product through a 40-mesh or 60-mesh sieve.

[0277] Process Aldavine™ 5X - A12 formulation

[0278] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0279] Add Sodium benzoate and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0280] Add Potassium sorbate and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0281] Add galactan syrup G6 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0282] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0283] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0284] Check pH (specification: 5.5-5.9), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0285] Filter the product through a 40-mesh or 60-mesh sieve.

[0286] Process Aldavine™ 5X - A13 formulation

[0287] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0288] Add Dermosoft™ 1388 ECO NAL and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0289] Add galactan syrup G7 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0290] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0291] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C. Check pH (specification: 5.5-5.9), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0292] Filter the product through a 40-mesh or 60-mesh sieve.

[0293] Process Aldavine™ 5X - A14 formulation

[0294] Shake the glycerin. Heat to 60+ / -3°C.

[0295] Add Softisan™ GC8 Palm Free and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0296] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 30 minutes or until the solution is homogeneous at 60+ / -3°C.

[0297] Add purified water. Shake for 5 minutes or until solution is homogeneous at 60+ / -3°C.

[0298] Add galactan syrup G8 (see Table 3). Shake for 5 minutes or until solution is homogeneous at 60+ / -3°C.

[0299] With rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0300] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0301] Filter the product through a 40-mesh or 60-mesh sieve.

[0302] Process Aldavine™ 5X - A15 formulation

[0303] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0304] Add A-Leen™ Aroma-3 and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0305] Add galactan syrup G9 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0306] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0307] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0308] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0309] Filter the product through a 40-mesh or 60-mesh sieve.

[0310] Process Aldavine™ 5X - A16 formulation

[0311] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0312] Add Dermosoft™ 1388 ECO NAL and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0313] Add galactan syrup G7 (see Table 3). Shake for 5 minutes at 60+ / -3°C. Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0314] Under vigorous agitation, add sorbitol powder. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0315] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0316] Filter the product through a 40-mesh or 60-mesh sieve.

[0317] Process Aldavine™ 5X - A17 formulation

[0318] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0319] Add Dermosoft™ 1388 ECO NAL and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0320] Add galactan syrup G7 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0321] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0322] Under vigorous agitation, add glycerin. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0323] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0324] Filter the product through a 40-mesh or 60-mesh sieve.

[0325] Process Aldavine™ 5X - A18 formulation

[0326] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0327] Add Dermosoft™ 1388 ECO NAL and shake for 5 minutes or until completely solubilized at 60+ / -3°C.

[0328] Add galactan syrup G7 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0329] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0330] Under vigorous agitation, add glycerin. Shake for a minimum of 2 hours or until solution is homogeneous at 60+ / -3°C.

[0331] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0332] Filter the product through a 40-mesh or 60-mesh sieve.

[0333] Process Aldavine™ 5X - A19 formulation

[0334] Heat purified water to 60+ / -3°C in the heated main tank fitted with a propeller mixer.

[0335] Add A-Leen™ Aroma-3. Shake for 5 minutes or until completely solubilized at 60+ / -3°C. Add galactan syrup G9 (see Table 3). Shake for 5 minutes at 60+ / -3°C.

[0336] Under rapid agitation, slowly add fucan. Shake vigorously for a minimum of 30 minutes or until completely solubilized at 60+ / -3°C.

[0337] Under vigorous agitation, add glycerin. Shake for a minimum of 2 hours or until the solution is homogeneous at 60+ / - 3°C.

[0338] Check pH (specification: 5.5-6.5), if out of specification, adjust with 1 N HCI or 2% NaOH and shake for a further 15 minutes.

[0339] Filter the product through a 40-mesh or 60-mesh sieve.

[0340] Sensitive Skin substitute model

[0341] For sensitive skin, substitutes were generated with skin cells (fibroblasts and keratinocytes), extracted from patient biopsies (Males or females, Caucasian, 36 to 65 years old).

[0342] Extracted fibroblasts were cultivated in DM EM supplemented with 10% fetal bovine serum (Hyclone), 100 U / mL penicillin G (Sigma) and 25pg / mL gentamicin (Shering), at 37°C under 5% CO2 and 95% humidity.

[0343] Extracted keratinocytes were cultivated in DMEM with Ham’s F12 (Gibco) at a 3 :1 ratio, supplemented with 5% Fetal Clone II serum (Hyclone), 10ng / mL epidermal growth factor EGF (Austral Biologicals), 100 U / mL penicillin G (Sigma), 25pg / mL gentamicin (Schering), 5pg / mL insulin (Sigma Aldrich), 0.4pg / mL hydrocortisone (Calbiochem) and 1010M cholera toxin (MP Biomedicals), at 37°C under 5% CO2 and 95% humidity.

[0344] After extraction and proliferation of the different skin cell types (fibroblasts and keratinocytes), the skin substitutes were reconstructed. Briefly, after 28 days of culture, fibroblasts were seeded in 6-well plates and then treated with ascorbic acid (50 pg / mL) to increase collagen production and to allow the superposition of two sheets to generate a surface equivalent to dermis. Then, keratinocytes were seeded on this dermal surface to produce an epidermis. After one week of culture, substitutes were elevated at the air-liquid interface to allow the keratinocytes differentiation and to obtain the different epidermal layers. 11 days later, the substitutes were treated, every 3 days, with Aldavine™ 5X (1 %, 2%) or Methotrexate (used as a positive control at 734 pM) in the culture medium for 10 days. At the end of the treatment, the culture of substitutes was stopped, and analysis were performed.

[0345] Masson's trichrome staining was performed to differentiate the different layers of the skin (i.e., stratum corneum, epidermis and dermis). Skin substitutes were fixed in formalin solution and embedded in paraffin. Masson's trichrome staining was then performed on 5 pm sections. This staining was done by a series of continuous baths of different products, i.e., toluene, Weigert's hematoxylin, water, fushin-ponceau, phosphomolybidine acid, ethanol, and aniline blue. Thicknesses of the epidermis were measured for each condition and the analysis was performed with Imaged™ software. Each condition was analyzed in triplicate.

[0346] Immunofluorescence Skin biopsies were included in Optimal Cutting Temperature (OCT) and then cut into thin 5 pm sections. The sections were fixed with acetone at -20 °C for 10 minutes and then washed. The slides were incubated with the primary antibody diluted in 1 % PBS-bovine serum albumin (BSA) solution for 45 minutes at room temperature or overnight in the dark at 4 °C. For Wnt5a, previously the staining, the cells were permeabilizated with Triton™ X100 solution. The primary antibodies used were anti-K14 antibodies (Cederlane), anti-Ki67 antibodies (Abeam), anti-loricrin antibodies (Biolegend) and anti-Wnt5a antibodies (Invitrogen). Washes were performed before re-incubating the slides in the dark for 20 minutes in the presence of secondary antibody. After incubation, three washes were performed before mounting the slides with DAPI Fluoromount-G™ (Southern Biotech). Pictures were taken under a confocal microscope (LSM 900 with Airscan 2, Carl Zeiss) or fluorescence microscope (Zeiss Axiolmager M2), and Imaged™ software was used to quantify the fluorescence.

[0347] Western blot

[0348] Skin tissues were collected and disrupted at -80°C using a cryomill (Vibrogrinder cryomill), the total proteins obtained were then lysed using the RIPA lysis buffer supplemented with proteinase inhibitor. Cell lysates were then centrifuged to remove cellular debris, and the clarified supernatants were used for western blot analysis. Proteins mixed with Laemmli buffer were separated by SDS-PAGE and then transferred to PVDF membranes. The membranes were blocked for 1 h with 5% non-fat powder milk in Tris-buffered saline (TBS)-0.05% Tween™ 20 (TBS-T) at room temperature and incubated with anti-Clasp2 primary antibody (Santa cruz) overnight (4°C). The next day, the membranes were incubated with HRP-conjugated anti-rabbit IgG secondary antibody for 1 h at room temperature. Immunoreactivity was visualized by enhanced chemiluminescence detection (ECL, Cytiva) and acquisitions were analyzed using Imaged™ software. test

[0349] The efficacy of various antimicrobial preservative agents was tested according to European Pharmacopoeia chapter

[0350] 5.1.3 11th edition.

[0351] Statistical

[0352] All data were analyzed using GraphPad™ Prism 9 software. Every experiment was repeated three times at least, and the data were shown as mean ± standard error of mean. Obtained data and percentage variations were submitted to one-way ANOVA test followed by a post-hoc test specific to each analysis. The statistical value is p<0.05 (*p<0.05,

[0353] **p<0.01, ***p<0.001).

[0354] EXAMPLE 2: Effect of Aldavine™ 5X treatment on the epidermis thickness.

[0355] Aldavine™ 5X at the 1 % and 2% concentrations was applied on skin substitute, and epidermis thickness measured as described in Example 1 .

[0356] As shown in FIG. 1 , Aldavine™ 5X significantly decreased epidermis thickness at both the 1 % and 2% concentrations tested by 33% and 40%, respectively. This decreased epidermis thickness denotes a decrease of skin cell proliferation. Obtained data were submitted to one-way ANOVA test followed by Kruskal-Wallis. EXAMPLE 3: Effect of Aldavine™ 5X treatment on the Ki67 expression.

[0357] Aldavine™ 5X at the 1 % and 2% concentrations was applied on skin substitute, and the expression of ki67, used as a marker of proliferation, was measured as described in Example 1.

[0358] As shown in FIG. 2, Aldavine™ 5X significantly decreased Ki67 expression at both the 1 % and 2% concentrations tested by 55% and 63%, respectively. This decreased ki67 expression denotes a decrease of skin cell proliferation. Obtained data were submitted to one-way ANOVA test followed by Kruskal-Wallis.

[0359] EXAMPLE 4: Effect of Aldavine™ 5X treatment on the K14 expression.

[0360] Aldavine™ 5X at the 1 % and 2% concentrations was applied on skin substitute, and the expression of K14, used as a marker of proliferation, was measured as described in Example 1 .

[0361] As shown in FIG. 3, Aldavine™ 5X significantly decreased K14 expression at 1 % by 19%. This decreased K14 expression denotes a decrease of skin cell proliferation. Obtained data were submitted to one-way ANOVA test with Dunnett's correction.

[0362] EXAMPLE 5: Effect of Aldavine™ 5X treatment on CLASP2 expression.

[0363] Aldavine™ 5X at the 1% and 2% concentrations was applied on skin substitute, and the expression of Clasp2, used as a marker of skin cell proliferation and migration, was measured as described in Example 1 .

[0364] As shown in FIG. 4, Aldavine™ 5X significantly decreased Clasp2 expression at both the 1 % and 2% concentrations tested by 44% and 86%, respectively. This decreased Clasp2 expression denotes a decrease in skin cell migration and proliferation. Obtained data were submitted to one-way ANOVA test followed by Kruskal-Wallis.

[0365] EXAMPLE 6: Effect of Aldavine™ 5X treatment Wnt5a expression

[0366] Aldavine™ 5X at the 1% and 2% concentrations was applied on skin substitute, and the expression of Wnt5a, used as a marker of skin cell proliferation and migration, was measured as described in Example 1 .

[0367] As shown in FIG. 5, Aldavine™ 5X significantly decreased Wnt5a expression at both the 1% and 2% concentrations tested by 17% and 22%, respectively. This decreased Wnt5a expression denotes a decrease in skin cell migration and proliferation. Obtained data were submitted to one-way ANOVA test with Brown-Forsythe 's correction.

[0368] EXAMPLE 7: Effect on differentiation Effect of Aldavine™ 5X treatment Loricrin expression

[0369] Aldavine™ 5X at the 1 % and 2% concentrations was applied on skin substitute, and the expression of loricrin, used as a marker of skin cell differentiation, was measured as described in Example 1 .

[0370] Aldavine™ 5X significantly increased Loricrin expression at both the 1% and 2% concentrations tested by 40% and 152%, respectively. This restored expression of loricrin indicates normal differentiation of skin cells. Obtained data were submitted to one-way ANOVA test followed by Kruskal-Wallis.

[0371] EXAMPLE 8: Challenge test

[0372] The efficacy of various antimicrobial preservation agents was tested as described in Example 1 . Microorganisms used were Staphylococcus aureus ATCC6538 for Gram+ bacteria, Pseudomonas aeruginosa ATCC 9027 and Escherichia coliMCC 8739 for Gram- bacteria, Candida albicans ATCC 10231 foryeasts and Aspergillus brasiliensis ATCC 16404 for molds.

[0373] EXAMPLE 9: Stability tests - UV spectrum and organoleptic properties The formulations and galactan syrup sent for organoleptic stability testing were divided into three samples. One sample was stored at 4°C, one at 25°C, and the last one at 50°C. These three samples were monitored at different time points: TO, 15 days (D15), 1 month (1 M), 3 months (3 M), and 6 months (6 M). At each time point, the parameters were monitored by a specialist technician who checks the maintenance of the various characteristics over time. The parameters monitored are odor, visual aspects (color, turbidity, sediment), and the UV spectrum. These parameters must remain stable at least three months or more relative to TO and also compared to the sample stored at 4°C to be considered stable. The assay was terminated at the initial time point when the formulation or galactan syrup demonstrated instability concerning at least one of the aforementioned organoleptic properties.

[0374] EXAMPLE 10: Effect of various preservative agents on formulation properties

[0375] The Table 5 below summarizes challenge test and stability results obtained for formulations A1-A19. The stability of specific galactan syrups (see Table 3 above) used in certain formulations is also presented in Table 5 below.

[0376] [Table 5]: Stability results

[0377]

[0378] The scope of the claims should not be limited by the embodiments set forth in the examples but should be given the broadest interpretation consistent with the description as a whole.

[0379] REFERENCES

[0380] Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. Colorimetric method for Determination of sugars and related substances. Anal Chem 1956;28(3):350-6.

[0381] Blumenkrantz N, Asboe-Hansen G. New method for quantitative determination of uronic acids. Anal Biochem 1973;54(2):484-9. [PubMed: 4269305] Nardella A, Chaubet F, Boisson-Vidal C, Blondin C,

[0382] Durand P, Jozefonvicz J. Anticoagulant low molecular weight fucans produced by radical process and ion exchange chromatography of high molecular weight fucans extracted from the brown seaweed Ascophyllum nodosum. Carbohydr Res 1996;289:201-8. [PubMed: 8805780]

[0383] Mulloy B, Gee C, Wheeler SF, Wait R, Gray E, Barrowcliffe TW. Molecular weight measurements of low molecular weight heparins by gel permeation chromatography. Thromb Haemost 1997;77(4):668-74. [PubMed: 9134640]

Claims

CLAIMS1 . Use of a topical composition comprising fucan and galactan in a dry weight ratio of between about 0.75 : about 1 (w / w) and about 20 : about 1 (w / w), water, a first preservative agent comprising at least 3% w / w glycerin, and a second preservative agent comprising phenylpropanol, to decrease epidermis thickness and / or skin cell proliferation in a subject in need thereof.

2. The use of claim 1 , wherein the composition comprises fucan : galactan in a dry weight ratio of between about 12 : about 1 (w / w) and about 18: about 1 (w / w).

3. The use of claim 1 or 2, wherein the fucan is extracted from brown algae and / or galactan is extracted from red algae.

4. The use of any one of claims 1-3, wherein the fucan is extracted from Ascophyllum nodosum and / or the galactan is extracted from Asparagopsis armata.

5. The use of any one of claims 1-4, wherein the second preservative agent consists of phenylpropanol.

6. The use of any one of claims 1-5, wherein the second preservative consists of about 0.2 to 0.4 % w / w phenylpropanol.

7. The use of any one of claims 1-6, wherein the first preservative agent comprises or consists of between about 14% w / w and about 65% w / w of glycerin.

8. The use of any one of claims 1-7, wherein the first preservative agent comprises or consists of between about 20% w / w and about 60% w / w of glycerin, or between about 30% w / w and about 60% w / w of glycerin, or between about 40% w / w and about 60% w / w of glycerin, or between about 45% w / w and about 55% w / w of glycerin, or between about 50% w / w and about 53% w / w of glycerin.

9. A stable topical composition comprising fucan and galactan in a dry weight ratio of between about 0.75 : about 1 (w / w) and about 20 : about 1 (w / w), water, a first preservative agent comprising between about 14% w / w and about 65% w / w glycerin, and a second preservative agent that comprises between about 0.2 to 0.4 % w / w phenylpropanol.

10. The composition of claim 9, comprises fucan : galactan in a dry weight ratio of between about 12 : about 1 (w / w) and about 18: about 1 (w / w).11 . The composition of claim 9 or 10, wherein the fucan is extracted from brown algae and / or galactan is extracted from red algae.

12. The composition of any one of claims 9-11 , wherein the fucan is extracted from Ascophyllum nodosum and / or the galactan is extracted from Asparagopsis armata.

13. The composition of any one of claims 9-12, wherein the second preservative agent consists of phenylpropanol.

14. The composition of any one of claims 9-12, wherein the second preservative consists of about 0.2 to 0.4 % w / w phenylpropanol.

15. The composition of any one of claims 9-14, wherein the first preservative agent comprises or consists between about 20% w / w and about 60% w / w of glycerin, or between about 30% w / w and about 60% w / w of glycerin, or between about 40% w / w and about 60% w / w of glycerin, or between about 45% w / w and about 55% w / w of glycerin or between about 50% w / w and about 53% w / w of glycerin.