Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai
Reexamination Certificate
2000-01-27
2003-01-14
Low, Christopher S. F. (Department: 1653)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Peptide containing doai
C514S002600, C514S015800, C514S016700, C514S018700, C514S019300, C530S360000, C530S361000
Reexamination Certificate
active
06506732
ABSTRACT:
BACKGROUND OF THE INVENTION
(a) Field of the Invention
The present invention relates to an hydrolysate of milk proteins containing a mixture of hydrophobic and hypoallergenic peptides having molecular weights and hydrophobicity suitable for effective penetration through proteino-lipidic layers of skin, in order to join and to feed the cells of the basal membrane of the epidermis and fibroblasts cells of the dermis.
(b) Description of Prior Art
British Patent application No. 2,046,591 discloses the utilization of whey protein hydrolysate as the active ingredient in skin care composition. The hydrolysate of whey proteins from milk is obtained by the action of an endopeptidase and it consists essentially of peptides having molecular weight in the range from 200 to 5000 daltons. This composition is only suitable for the hydration of skin. It is also known to use casein hydrolysate as hair and skin treating agent. The hydrolysate of casein proteins from milk is obtained by the action of an endopeptidase and a bacterial protease, and it consists of peptides having molecular weight in the range from 300 to 3000 daltons. This agent is suitable in skin care composition for moisturizing property and protective property.
In the prior art referred above, the effectiveness of the peptides to penetrate skin and to promote cell growth, regeneration and repair has not been proven.
U.S. Pat. No. 5,314,873 reveals the utilization of milk proteins hydrolysates, including casein hydrolysate and fraction obtained from such hydrolysate, in cosmetic compositions to activate growth of human cutaneous cells. Hydrolysates are said to contain peptides lower than 1000 daltons and fraction is said to contain aromatic amino acids in an amount of less than 5% by weight of total amino acids and to have proliferation activating property on human cutaneous cells but does not have the antigenicity of the milk protein. Casein was hydrolyzed with a microbial enzyme from Aspergilus and a mixture of pancreatic enzymes to a decomposition level of 41%. Growth of human skin cells, cultured in KGM and KGM modified culture media containing 2% of bovine fetal serum and 100&mgr;g/ml of hydrolysate or fraction of hydrolysate, was used to measure proliferation activity. Proliferation activity of hydrolysate and fractions of hydrolysates was found to be 7 and 13% respectively for keratinocytes, and 4 and 31% respectively for fibroblasts. Antigenicity was determined by ELISA inhibition test using rabbit antiserum and goat anti-rabbit antibody. In the prior art referred above, the effectiveness of peptides to penetrate skin and to promote cells growth has not be proven, as well as the allergenicity in humans and wound healing.
U.S. Pat. No. 5,166,132 reveals that a composition containing casein hydrolysate, carrageenan, poly-vinyl pyrrolidone, methyl paraben or propyl paraben, has wound healing property.
Normal undamaged skin is composed of several layers. The outer layer, usually called the epidermis, is composed of several types of epithelial cells, nerve fibrils, but does not contain blood vessels. Cells that are at or near the base of the epidermis, near the dermo-epidermial junction, are called basal cells. The basal cells generate new keratinocytes that progressively differentiate while migrating to the surface of the skin. During that process, cells produce keratin, a highly resistant fibrous protein that form a solid and water-repellent matrix by interaction with the other structural proteins and lipids of the cells. Below the epidermis is a layer of cells and connective tissue called the dermis. This layer comprises lymph and blood vessels that feed the cells of the epidermis and dermis. Also, hair follicles, sebaceous glands and sweat glands extend from the dermis to the surface of the skin. Fibroblasts cells in the dermis produce collagen, a fibrous protein that forms a structural matrix which increases the strength of the tissue. The proliferation of fibroblasts and the production of collagen are very important during the wound-healing process.
In vitro cell culture from a patient own skin is now used to produce new skin layers or autografts for the treatment of severely burned patients and those suffering important skin loss. Culture media used must contain appropriate nutrients and growth factors such as Epidermal Growth Factor (EGF) to provide rapid growth of undifferentiated keratinocytes cells. Fetal calf serum (FCS), commonly used for that purpose as a source of protein nutrients and growth factors in undifferentiated culture media, is however very expensive. Fetal calf serum is also used in fibroblast culture media to promote cells growth.
It would be highly desirable to be provided with less expensive peptides from casein hydrolysates to replace FCS in undifferentiated culture media.
It would also be highly desirable to be provided with a milk protein hydrolysate or a fraction thereof which may be used in skin cell culture media, or in cosmetic or pharmaceutical formulations.
It would also be highly desirable to be provided. with a milk protein hydrolysate or a fraction thereof which may be used in cosmetic formulations to promote hypoallergenic, skin hydrating and skin regenerating properties.
It would also be highly desirable to be provided with a fraction of milk protein hydrolysate which may be used in skin cell culture media to promote proliferation of cells.
It would also be highly desirable to be provided with a fraction of milk protein hydrolysate which may be used in pharmaceutical preparations to promote percutaneous absorption and wound healing properties.
It would also be highly desirable to be provided with a method for producing a milk protein hydrolysate or a fraction thereof.
SUMMARY OF THE INVENTION
One aim of the present invention is to provide a milk protein hydrolysate or a fraction thereof for use in skin cell culture media, or in cosmetic or pharmaceutical formulations.
Another aim of the present invention is to provide a milk protein hydrolysate or a fraction thereof which may be used in cosmetic formulations to promote hypoallergenic, skin hydrating and skin regenerating properties.
Another aim of the present invention is to provide a fraction of milk protein hydrolysate which may be used in skin cell culture media to promote proliferation of cells.
Another aim of the present invention is to provide a fraction of milk protein hydrolysate which may be used in pharmaceutical preparations to promote percutaneous absorption and wound healing properties.
Another aim of the present invention is to provide a method for producing a milk protein hydrolysate or a fraction thereof to attain the objects and advantages of the present invention.
In accordance with the present invention there is provided a milk protein hydrolysate comprising a first fraction containing a mixture of peptides having a molecular weight of about 900 daltons, ranging from 200 to 1400 daltons, and a hydrophobicity of about 11 Kcal/mole, and a second fraction containing a mixture of soluble hydrophobic peptides having a molecular weight of about 2000 daltons, ranging from 1400 to 2600 daltons. The mixture of peptides having a molecular weight of about 900 daltons contains a total hydrophobic amino acids concentration and an aromatic amino acids concentration of at least 65% and 17% respectively; the peptides have aromatic amino acids and other hydrophobic amino acids located at the C- and N-terminal ending in proportion over 85%.
Still in accordance with the present invention, there is provided an enzymatic hydrolysate of milk proteins comprising a mixture of soluble peptides comprising a first fraction containing peptides of a molecular weight ranging from 200 daltons to 1400 daltons and having average hydrophobicity of 11 Kcal/mole, a second fraction containing hydrophobic peptides of molecular weight ranging from 1400 to 2600 daltons. The hydrolysate has low allergenicity and skin hydration property. Preferred peptides of the present invention are, without limitation, selected from the group consisting of ALPQYL (SEQ
Low Christopher S. F.
Lukton David
Universite Laval
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