Process of making and using composition containing oxidation-sensitive hydrophilic active principle and maleic anhydride copolymer
a technology of hydrophilic active principle and composition, which is applied in the field of making and using compositions containing oxidation-sensitive hydrophilic active principles and maleic anhydride copolymers, can solve the problems of easy decomposition on contact with water, conflicting with the desired effect, and inability to be stable in aqueous medium,
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example 1
Accelerated Storage Test
[0086] The aim of this test is to study the decomposition of an oxidation-sensitive hydrophilic active principle after storing for two months at 45.degree. C. Various solutions were prepared and their compositions are collated in the following table:
1 TABLE I Compositions Ascorbic (in water) acid Polymer 1 Polymer 2 Solution A 15% -- --(Control 1) Solution B 15% 1% -- Solution C 15% -- 1% Solution D 5% -- -- (Control 2) Solution E 5% 1% -- Solution F 5% -- 1%
[0087] All the solutions are brought to pH 6 with 8.9 mol / l KOH.
[0088] The percentages of the polymers are given as active material.
[0089] Polymer 1: Styrene / maleic anhydride (50 / 50) copolymer, in the form of an ammonium salt at 30% in water, sold under the reference SMA1000H.RTM. by Atofina.
[0090] Polymer 2: Styrene / maleic anhydride (50 / 50) copolymer, in the form of a sodium salt, sold under the reference SMA1000HNa.RTM. by Atofina.
[0091] The degree of decomposition measured is given by the ratio:
(C.sub....
example 2
Effect of the Composition According to the Invention on the Differentiation of Keratinocytes
[0102] Principle of the Method:
[0103] The subject consists in evaluating the effect of the combination of ascorbic acid and of a copolymer according to the invention on the differentiation of keratinocytes by measuring the transglutaminase (TGk) activity of cultured human epidermal keratinocytes. The TGk is a marker of the terminal differentiation of the keratinocyte and of the formation of the cornified envelope (corneocyte).
[0104] Human epidermal keratinocytes used at the 3rd passage are inoculated in a 96-well plate at a density of 10 000 cells per well and are cultured in complete SFM medium (Gibco 170005034, EGF and pituitary extract). After preincubating for 24 hours, the cells are brought into contact with the product (tested at 30 .mu.M) for 48 hours. The cells are subsequently washed and then treated with ultrasound on ice in Tris / EDTA, pH 8, buffer. The membrane enzyme is extracted ...
example 3
Effect of the Composition According to the Invention on the Synthesis of Epidermal Ceramides
[0109] Protocol:
[0110] The study is carried out on a reconstructed epidermis in vitro after culturing for 7 days.
[0111] Before application of the combination of ascorbic acid and of a copolymer according to the invention, the reconstructed epidermis is incubated overnight with .sup.14C acetate (2 .mu.Ci / ml), in order to monitor and quantify the synthesis of the epidermal lipids synthesized during the contact period.
[0112] The test solutions are introduced between the 8th and 14th day of culturing.
[0113] The ascorbic acid concentration is 50 .mu.g / ml.
[0114] Analysis of the Lipids:
[0115] At the end of the incubation, the epidermis equivalent is detached to form its collagen support.
[0116] The extraction and the analysis of the epidermal lipids by HPTLC (high performance thin layer chromatography) are carried out according to the protocol described in M. Ponec (1991, Adv. Lipid Res., 24: 83-117)...
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