Field of the Invention

The present invention relates to a composition comprising whole plant extract of Acanthophora spicifera as an anti-aging agent. The present invention also relates to a composition comprising the whole plant extract of Acanthophora spicifera as UV protection agent.

Background and the Prior Art of the Invention

Skin aging is of two types- chronological aging and photo aging. Skin aging can be addressed through various mechanisms such as
1) Anti-oxidants (for protection from damaging effects caused by free radicals)
2) Activators of collagen synthesis and/or
3) MMP inhibitors (to decrease break-down of collagen)

Firstly, free radicals generated in the skin leads MMP synthesis which is the enzyme that degrades the extra cellular matrix protein, which is mainly collagen. This leads to changes in the skin structure contributing to aging. Secondly, when anti-oxidants are used, they scavenge the free radicals formed due to photo damage and prevent the synthesis of enzymes that degrade the collagen.

MMP Inhibitors are Matrix metalloprotease inhibitors which inhibit the Matrix metalloproteins, that from a part of the extra cellular matrix in the skin fibroblast, thereby disturbing the skin integrity and firmness. The role of MMP inhibitors is to inhibit the enzyme activity of MMPs thereby preventing collagen degradation.

Ganesan et al., “Antioxidant of the Methanol Extract and its Soluble Fractions obtained from selected Indian Seaweeds” Bioresource technology; Volume 99, Issue 8, May 2008, Pages 2717-2723 discloses in vitro antioxidant activity of crude methanolic extracts of Acanthophora sp. along with 2 other seaweeds. The ethyl acetate fraction of Acanthophora methanolic extract was found to exhibit higher antioxidant activity. This document demonstrates that the selected seaweeds exhibit anti-oxidant activity.

The study of Hannah et al; “Hepatoprotective and Antioxidant Activity of the Marine Red Algae Acanthophora spicifera on CCl4 Induced Liver Toxicity in Rats” Journal of the Indian Society of Toxicology, Volume 2, Issue 1, 2006 shows the hepatoprotective and antioxidant effect of Acanthophora spicifera in CCl4 intoxicated male albino rats. Ethanolic extract of Acanthophora could decrease the level of lipid peroxidation and increase the level of antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase. It also exhibited liver protection function. The biological activity of the extract was related to the phyto-constituents such as flavanoids, Vitamins -A, E and C present in the algae.
Han et al; “Bioactive sterols from red algae Acanthophora spicifera” Zhongguo Zhong Yao Za Zhi. Volume 34, Issue 1, Jan 2009, Pages 60-63, the chemical constituents of the red alga Acanthophora spicifera were studied for identifying new bio-actives. Six sterols were isolated from the ethanolic extract of Acanthophora sp. out of which three compounds showed moderate cytotoxic activity against human cancer cell line.
Zeng et al; “Flavanoids from the Red Algae Acanthophora spicifera” Chinese J. Chem 19; 2001. Two new flavanoids i.e., acanthophorin A and acanthophorin B were isolated from the ethanolic extracts of Acanthophora along with 3 known flavanoids such as catechin, quercetin and tiliroside. The study shows that the newly isolated flavanoids could prevent lipid peroxidation and inhibit the generation of malondialdehyde in liver homogenates of rat in vitro.
JP 2124810 discloses that to obtain the cosmetics such as foundations or hair cosmetics of high stability, skin color-lightening action and a suitable skin astringency by using the extracts from sea weeds of brown algae such as hizikia, sea mustard or sea oak with a hydrophilic solvent. The composition of the extract comprises of a sea weed of brown algae, preferably tangle or gulfweed is extracted with a hydrophilic solvent such as methanol or ethanol and the extract is added to prepare the subject cosmetic. The hydrophilic solvent is preferably used in the single form of an alcohol of 1 to 6 carbon atom or a mixture thereof with water. The content of the extract in the cosmetic is 0.005 to 3 wt %, preferably 0.05 to 0.3 wt %. The cosmetic has adequately tyrosinase- inhibiting action in a low concentration in the same order as that of L-ascorbic acid and also a proper level of protein coagulation, having high stability.
JP2001302492 provide a novel and safe melanin formation inhibitory agent and a cosmetic composition containing the same. The objective melanin formation inhibitory agent includes one kind or two or more kinds of seaweed extracts that are obtained from alkali- treated seaweeds in red algae and the objective cosmetic composition including the melanin formation inhibitory agent. The sea weed extracts have excellent melanin formation inhibitory action, alleviate skin stains and freckles, further have beautifully whitening effect, ameliorate roughened and dry skins and give the skin gloss and tension.

The available non-patent literature discloses the presence of flavanoids, anti-oxidants and sterols in Acanthophora sp. exhibiting moderate anti-cancer and hepatoprotective activities.

Mere scavenging free radicals using anti-oxidants alone do not induce the synthesis of collagen. Similarly, the use of MMP inhibitors could prevent collagen degradation but not activate collagen synthesis.

MMP Inhibitors are Matrix metalloprotease inhibitors which inhibit the Matrix metaloproteins that form a part of the extra cellular matrix in the skin fibroblast thereby disturbing the skin integrity and firmness. The use of anti-oxidants and/or MMP inhibitors in the prior art does not provide any lead to its role in collagen synthesis, which is advantageous for anti-aging.

Further, the prior art discloses various assays to demonstrate the presence of anti-oxidant activity in Acanthophora sp. However, none of the prior art targets the activation of collagen synthesis. Therefore there is a need to provide a skin care composition that can provide a better skin anti-aging effect.

Objects of the Invention
It is one of the objects of the present invention to provide the whole plant extract of Acanthophora spicifera extracts as an anti-aging component having a role on activation of collagen synthesis.

Yet another object of the present invention is to use the whole plant extract of Acanthophora spicifera for UV protection.

Summary of the invention
A skin care composition comprising Acanthophora spicifera extract along with conventional additives wherein the said extract is present in a range of 0.1 to 5%.
Brief Description of Accompanying Figures

Figure 1 illustrates collagen activation by cream containing aqueous extract of Acanthophora in fibroblast cells (in-vitro study).

Figure 2 illustrates the effect of cream containing Acanthophora extract on fibroblast cells to demonstrate collagen synthesis using Sircol collagen assay.

Figure 3 illustrates the effect of Acanthophora extract containing cream on the proliferation rate of fibroblast cells.

Figure 4 illustrates decrease in ROS by Acanthophora extracts related to demonstrate the UV protection activity.

Detailed Description of the Invention

The present invention targets the “activation of collagen synthesis” using the whole plant extract of Acanthophora spicifera regardless of the presence of free radicals in the system.

According to one embodiment of the present invention, a skin care composition is provided comprising the whole plant extract of Acanthophora spicifera extract along with other conventional additives.

The inventors of present invention have found that there is no significant activity of collagen synthesis below 0.125 ppm by using Acanthophora extract However, there is a significant increase in the collagen synthesis activity between 0.1 ppm and 5 ppm. In accordance with the present invention, Acanthophora extract is present in the topical composition in an effective amount ranging from 0.1 to 5% w/w, more preferably 0.1 to 3% w/w, most preferably 0.1 to 1% w/w, that shows significant increase in collagen, anti aging applications.

The skin care composition as referred herein comprises two phase system: a) water phase and b) oil phase.

The water phase of the skin care composition comprises xanthan gum, di sodium EDTA, magnesium sulphate, acrylates/C10-30 Alkyl Acrylate Crosspolymer and ammonium Acryloyl - dimethyltaurate / VP Copolymer.
The oil phase of the skin care composition comprises C12-15 Alkyl Benzoate, Potassium Cetyl Phosphate (and) Hydrogenated Palm Glycerides, Cetostearyl alcohol and Glyceryl Stearate and PEG-100 Stearate.
The invention provides the skin care composition in the form of a lotion, cream or gel. The conventional additives of the present skin care composition are selected from preservatives, rheology modifiers, neutralizing agents or fragrances.
Preservatives are selected from phenoxyethanol and ethylhexyl glycerin.
Rheology modifiers are selected from Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate Copolymer (and) Isohexadecane (and) Polysorbate 60.

Neutralizing agents or fragrances are selected from Triethanolamine, Fragrance- Unique D0803452.
One of the objectives of the present invention is to use Acanthophora extract at a concentration of preferably about 0.1 to 5% by weight, more preferably 0.1 to 3% and most preferably 0.1 to 1% by weight in the skin care composition.

The present invention further embodies the Collagen Synthesis Assay (Dye Binding Assay) using the reagents Fibroblast cell culture, DMEM cell culture media, Ammonium Sulphate and Sircol reagent. The reagent used for extraction is distilled water.

Detailed description of the accompanying examples:
Figure 1 represents the effect of Acanthophora extract containing cream (Aca Cream at different concentrations) on collagen synthesis in fibroblast cells.
Figure 2 represents the effect of fibroblast cell proliferation due to the addition of base cream and Acanthophora containing cream formulations.

Figure 3 represents the effect of Acanthophora spicifera extract in preventing the formation of Radical Oxygen Species (ROS) generation in fibroblast cells post UV exposure.

The present invention is illustrated by the following non-limiting examples. It is to be understood that the disclosed methodology is not limited to the exact details briefed here and variations to implement the idea are possible. The methodology described is for the purpose of description and should not be taken as limitation.

Example 1: Extraction Procedure details:

10% of the solvent extraction of the powdered algae material was carried out using water and the solvent was evaporated in a vacuum evaporator to obtain the extract in a powder form which was further used for the study.

Example 2: Preparation of skin care cream composition

Table 1: Preparation of Acanthophora extracts containing skin care cream composition

S.No. INCI Name Trade Name % w/w Function
1 Pasteurized water* Pasteurized water* 85.18
2 Ammonium Acryloyl - dimethyltaurate / VP Copolymer Aristoflex AVC
13824023147 0.05 Water phase
3 Acrylates/C10-30 Alkyl Acrylate Crosspolymer Pemulen TR2 0.17 Water phase
4 Xanthan gum Xanthan Gum Xanoluc FF Pharma Grade 0.10 Water phase
5 Disodium EDTA Disodium EDTA 0.10 Water phase
6 Magnesium sulfate Magnesium sulfate 0.70 Water phase
7 C12-15 Alkyl Benzoate Crodamol AB 3.00 Oil phase
8 Potassium Cetyl Phosphate (and) Hydrogenated Palm Glycerides Emulsiphos (677660) 2.00 Oil phase
9 Cetostearyl alcohol Ginol 16-18 TA 1.50 Oil phase
10 Glyceryl Stearate (and) PEG-100 Stearate Arlacel 165
PCM680160 2.00 Oil phase
11 Triethanolamine Triethanolamine 99% 0.20 Neutralizer
12 Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate Copolymer (and) Isohexadecane (and) Polysorbate 60 Simulgel INS 100 3.00 Rheology modifier
13 Acanthophora spicifera extract 1 Active
14 Phenoxyethanol Phenoxetol
17109122050 0.70 Preservatives
15 Ethylhexyl Glycerin Sensiva SC 50 0.50 Preservatives
16 Fragrance Fragrance Unique D0803452 0.80 Fragrance

Example 3: Preparation of skin care spray gel composition containing Acanthophora extracts

Table 2: Preparation of skin care spray gel composition

S.No. INCI Name Trade Name % w/w Function
1 Pasteurized water* Pasteurized water* 98.2
2 AmmoniumAcryloyl dimethyltaurate/VP Copolymer Aristoflex AVC 0.5 Polymer
3 Phenoxyethanol Phenoxetol
17109122050 0.3 Preservative
4 Acanthophora spicifera extract 1 Active

Example 4: Comparative data demonstrates the activity of 0.1 to 5% by weight of Acanthophora extract.

The cream composition contains about 0.1 to 5% by weight of Acanthophora extract. In vitro study of Acanthophora extract reveals increased collagen synthesis at 0.1 to 5ppm, however, beyond the said range Acanthophora extract does not induce collagen. To demonstrate the same (negative test results), 1% Aca cream was diluted to a concentration of 0.01 to 10 ppm to test the collagen synthesis under in-vitro conditions, the data has been provided in figure 2.

Example 5: Method of preparing skin care cream composition containing the Acanthophora extracts:

a) Weigh the water phase and heat upto 75 to 80˚C –Phase A
b) Weigh the oil phase and heat upto 75 to 80˚C –Phase B
c) When both phase A and B reaches 80˚C, phase B is mixed with phase A in a homogenizer for 15 to 20minutes.
d) The cream is then allowed to cool, when the temperature reaches 60˚C rheology modifiers and preservatives are added.
e) Further when the temperature reaches 40˚C, A. spicifera extract and fragrance is added and mixed well.

Example 6: Effect of cream and gel containing Acanthophora extract containing on fibroblast cells to demonstrate collagen synthesis using Sircol collagen assay as shown in Figure 1:
1. Fibroblast cells were treated with Acanthophora spicifera containing cream (Aca cream). The amount of collagen synthesized was measured.
2. Aca cream could induce 69% excess collagen at a concentration of 0.1ppm.

Fibroblast cells were treated with placebo/base formulations and Acanthophora extract containing formulations. Percent collagen synthesized at various cell treatment conditions and cells without any treatment (IC) was measured using Sircol collagen dye assay. Increased collagen synthesis was obtained with the addition of 0.1,1 and 5 ppm of Aca cream. No increase in collagen synthesis was observed below and above this range.
The terms as referred in figure 1 are defined below:
• IC-Inoculum control fibroblast cells without any treatment
• Base cream – Fibroblast cells treated with a base cream without Acanthophora extract
• 0.01, 0.1, 1, 5 and 10ppm Aca Cream – Fibroblast cells treated with Acanthophora extract cream at 0.01, 0.1, 1, 5 and 10ppm concentrations

Example 7: Effect of cream containing Acanthophora extract on the proliferation rate of fibroblast cells as shown in Figure 2.

1. Proliferation rate of fibroblast post treatment of the cell with Aca cream at varying concentrations was tested using MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assay.
2. Aca cream increased fibroblast proliferation by 26% at 1 ppm concentration.

Figure 2 represents the effect of fibroblast cell proliferation due to the addition of base cream and Acanthophora containing cream formulations. Percent fibroblast proliferation rate of the untreated control (IC), base cream and Aca cream was obtained using MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay. Maximum increase in fibroblast proliferation (26% more than untreated control cells) was observed in cells treated with 1ppm of Aca cream. Followed by 5, 10 ppm extract containing cream with proliferation rate of 25.6, 20% increase more than untreated control cells.

Example 8: Percent decrease in Radical Oxygen Species (ROS) generation post Acanthophora spicifera treatment as shown in Figure 3.

1. Fibroblast cells (both untreated control and Acanthophora spicifera extract treated) were exposed to stress (UV exposure) and level of ROS generated was measured.
2. Acanthophora spicifera treated cells produced less ROS when compared with the untreated cells and ascorbic acid treated cells.

Figure 3 represents the effect of Acanthophora spicifera extract in preventing the formation of Radical Oxygen Species (ROS) generation in fibroblast cells post UV exposure. Acanthophora spicifera extract at a concentration of 10 and 1 ppm decreased ROS generation by 37 and 20% respectively when compared to the control untreated cells. Ascorbic acid, a well-known antioxidant inhibited ROS generation by 14% (10ppm) and 28% (1ppm).

WE CLAIM:

1. A skin care composition comprising Acanthophora spicifera extract along with conventional additives wherein the said extract is present in a range of 0.1 to 5% by weight.
2. The skin care composition as claimed in claim 1, wherein said extract is present in a range of 0.1 to 3% by weight, more preferably 0.1 to 1% by weight of Acanthophora extracts.
3. The skin care composition as claimed in claim 1, wherein said skin care composition comprises a two phase system: a) water phase and b) an oil phase.
4. The skin care composition as claimed in claim 3, wherein said water phase comprises xanthan gum, di sodium EDTA, magnesium sulphate, acrylates/C10-30 Alkyl Acrylate Crosspolymer and ammonium Acryloyl - dimethyltaurate / VP Copolymer.
5. The skin care composition as claimed in claim 3, wherein said oil phase comprises C12-15 Alkyl Benzoate, Potassium Cetyl Phosphate (and) Hydrogenated Palm Glycerides, Cetostearyl alcohol and Glyceryl Stearate and PEG-100 Stearate.
6. The skin care composition as claimed in claim 1, wherein Acanthophora spicifera extract is from whole plant.
7. The skin care composition as claimed in claim 1, wherein Acanthophora spicifera extract is an aqueous extract.
8. The skin care composition as claimed in claim 1, wherein the composition is a lotion, cream, spray or gel.
9. The skin care composition as claimed in claim 1, wherein the additives are selected from preservatives, rheology modifier, neutralizer or fragrance.
10. The skin care composition as claimed in claim 1, wherein the Acanthophora extracts acts as activators of collagen synthesis.
11. The skin care composition as claimed in claim 1, wherein the composition is an anti-aging composition.
12. The skin care composition as claimed in claim 1, wherein the composition acts as a UV protective agent.

ABSTRACT

A skin care composition comprising Acanthophora spicifera extract along with conventional additives wherein the said extract is present in a range of 0.1 to 5% by weight.