Anti-Ageing Cosmetic Compositions for Men and Women, Containing Bioactive Protein, and the Method of its Production
The present invention relates to anti-aging composition comprising bioactive protein obtained by cultivation of placenta, method of preparing the composition and use as a cosmetic composition.
1 : Anti-ageing cosmetic compositions for men and woman comprising bioactive protein, which are obtained by the method of cultivation of explants XX-placenta (for women's cosmetics) or XY-placenta (for men's cosmetics) comprises the steps of: i. Making explants of XX-placenta or XY-placenta in gestation term not less 11 weeks, and not more 12 weeks. ii. Cultivation explants of XX-placenta or XY-placenta until the moment of migration and proliferation of cells on an area of not more than 4 cm 2 around each explant. iii. Filtration of the conditioned medium of XX-placenta or XY-placenta explants and adding a proteolysis inhibitor to the medium. iv. Control of biological safety of the conditioned medium of XX-placenta or XY-placenta explants (microbial, viral and fungal contamination). v. Controlling the concentration of biologically active proteins in a conditioned medium of XX-placenta or XY-placenta explants. vi. Adding of XX- or XY-placental compositions of biologically active proteins to the ingredients of cosmetic products. vii. Controlling the concentration of biologically active proteins of the XX-placenta or XY-placenta in ready-to-use cosmetic products.
2 : The method of making explants of XX-placenta or XY-placenta according to claim 1 ; wherein explants production performed after medical pregnancy termination in the gestational period 11-12 weeks.
3 : The method for obtaining biologically active proteins in a conditioned medium of XX-placenta or XY-placenta explants according to claim 1 ; wherein cultivation of XX-placenta or XY-placenta explants stops immediately after migration and proliferation of placental cells in an area of up to 4 cm 2 around each explant.
4 : The method as claimed in claim 1 , wherein conditioned medium of XX-placenta or XY-placenta explants is cleared of tissue and cells of placental explants by filtration.
5 : The method as claimed in claim 1 , wherein the degradation of biologically active proteins is prevented by the adding of the proteolysis inhibitor aprotinin to the conditioned medium of XX-placenta or XY-placenta explants.
6 : The method as claimed in claim 1 , wherein control of biological safety of the conditioned medium of XX-placenta or XY-placenta explants (microbial, viral and fungal contamination) is carried out according to the guidelines for monitoring the biological safety of stem cells and their derivatives.
7 : The method as claimed in claim 1 , wherein for control the concentrations of biologically active proteins in a conditioned medium of XY-placenta explants perfumed ELISA-test of references levels for: EGF—from 10 to 18 pg/ml; FGF-7—from 74 to 300 pg/ml; HGF—from 941 to 2039 pg/ml; TNF-alpha—from 340 to 521 pg/ml; IGF-1—from 630 to 928 ng/ml; VEGF-A—from 12 to 38 ng/ml; SCF—from 15.5 to 24 ng/ml; TGF-beta 1—from 66.5 to 178 ng/ml; ANGPT1—from 94 to 170 ng/ml; bFGF—from 58 to 97 ng/ml.
8 : The method as claimed in claim 1 , wherein for control the concentrations of biologically active proteins in a conditioned medium of XX-placenta explants perfumed EFISA-test of references levels for: EGF—from 8 to 18 pg/ml; FGF-7—from 90 to 265 pg/ml; HGF—from 30 to 104 pg/ml; TNF-alpha—from 340 to 546 pg/ml; IGF-1—from 514 to 681 ng/ml; VEGF-A—from 16 to 39 ng/ml; SCF—from 7.5 to 12 ng/ml; TGF-beta 1—from 311 to 550 ng/ml; ANGPT1—from 104 to 156 ng/ml; bFGF—from 14 to 43 ng/ml.
9 : The method as claimed in claim 1 , wherein adding of XX- or XY-placental compositions of biologically active proteins to the ingredients of cosmetic products, which are compatible with biologically active proteins and do not destroy them.
10 : The method as claimed in claim 1 , wherein controlled concentrations of biologically active proteins of the XY-placenta in ready-to-use cosmetic products must be not less then for: EGF—1.0 pg/ml; FGF—7-7.4 pg/ml; HGF—94.1 pg/ml; TNF-alpha—34.0 pg/ml; IGF-1—63.0 ng/ml; VEGF-A—1.2 ng/ml; SCF—1.6 ng/ml; TGF-beta 1—6.7 ng/ml; ANGPT1— 9.4 ng/ml; bFGF—5.8 ng/ml.
11 : The method as claimed in claim 1 , wherein controlled concentrations of biologically active proteins of the XX-placenta in ready-to-use cosmetic products must be not less then for: EGF—0.8 pg/ml; FGF—7-9.0 pg/ml; HGF—3.0 pg/ml; TNF-alpha—34.0 pg/ml; IGF-1—51.4 ng/ml; VEGF-A—1.6 ng/ml; SCF—0.8 ng/ml; TGF-beta 1—31.1 ng/ml; ANGPT1— 10.4 ng/ml; bFGF—1.4 ng/ml.
12 : The method as claimed in claim 3 , wherein conditioned medium of XX-placenta or XY-placenta explants is cleared of tissue and cells of placental explants by filtration.
13 : The method as claimed in claim 4 , wherein the degradation of biologically active proteins is prevented by the adding of the proteolysis inhibitor aprotinin to the conditioned medium of XX-placenta or XY-placenta explants.
14 : The method as claimed in claim 3 , wherein control of biological safety of the conditioned medium of XX-placenta or XY-placenta explants (microbial, viral and fungal contamination) is carried out according to the guidelines for monitoring the biological safety of stem cells and their derivatives.
15 : The method as claimed in claim 6 , wherein for control the concentrations of biologically active proteins in a conditioned medium of XY-placenta explants perfumed ELISA-test of references levels for: EGF—from 10 to 18 pg/ml; FGF-7—from 74 to 300 pg/ml; HGF—from 941 to 2039 pg/ml; TNF-alpha—from 340 to 521 pg/ml; IGF-1—from 630 to 928 ng/ml; VEGF-A—from 12 to 38 ng/ml; SCF—from 15.5 to 24 ng/ml; TGF-beta 1—from 66.5 to 178 ng/ml; ANGPT1—from 94 to 170 ng/ml; bFGF—from 58 to 97 ng/ml.
16 : The method as claimed in claim 6 , wherein for control the concentrations of biologically active proteins in a conditioned medium of XX-placenta explants perfumed EFISA-test of references levels for: EGF—from 8 to 18 pg/ml; FGF-7—from 90 to 265 pg/ml; HGF—from 30 to 104 pg/ml; TNF-alpha—from 340 to 546 pg/ml; IGF-1—from 514 to 681 ng/ml; VEGF-A—from 16 to 39 ng/ml; SCF—from 7.5 to 12 ng/ml; TGF-beta 1—from 311 to 550 ng/ml; ANGPT1—from 104 to 156 ng/ml; bFGF—from 14 to 43 ng/ml.
17 : The method as claimed in claim 6 , wherein adding of XX- or XY-placental compositions of biologically active proteins to the ingredients of cosmetic products, which are compatible with biologically active proteins and do not destroy them.
18 : The method as claimed in claim 9 , wherein controlled concentrations of biologically active proteins of the XY-placenta in ready-to-use cosmetic products must be not less then for: EGF—1.0 pg/ml; FGF—7-7.4 pg/ml; HGF—94.1 pg/ml; TNF-alpha—34.0 pg/ml; IGF-1—63.0 ng/ml; VEGF-A—1.2 ng/ml; SCF—1.6 ng/ml; TGF-beta 1—6.7 ng/ml; ANGPT1— 9.4 ng/ml; bFGF—5.8 ng/ml.
19 : The method as claimed in claim 9 , wherein controlled concentrations of biologically active proteins of the XX-placenta in ready-to-use cosmetic products must be not less then for: EGF—0.8 pg/ml; FGF—7-9.0 pg/ml; HGF—3.0 pg/ml; TNF-alpha—34.0 pg/ml; IGF-1—51.4 ng/ml; VEGF-A—1.6 ng/ml; SCF—0.8 ng/ml; TGF-beta 1—31.1 ng/ml; ANGPT1— 10.4 ng/ml; bFGF—1.4 ng/ml.