Claims for Patent: 5,891,426

Summary for Patent: 5,891,426

Title:	Screening of candidates for biological hair care activity
Abstract:	The invention concerns a procedure for testing a potentially-active substance used in the hair-care field, this procedure being characterized by the fact that a fragment of at least one hair follicle without dermal papilla is isolated, this fragment being taken from beneath the point of attachment of the sebaceous gland and excluding the portion of the hair shaft located above the point of attachment of the sebaceous gland; that this fragment is incubated in a suitable culture medium for a sufficient period; that this fragment is placed in contact with a potentially-active hair-care substance; that a marker of the activity of said tested substance is quantitatively determined; and that the results of this determination are assessed as they compare to a control.
Inventor(s):	Jarrousse; Fran.cedilla.oise (Livry Gargan, FR), Commo; Stephane (Paris, FR), Gaillard; Olivier (Paris, FR), Bernard; Bruno (Neuilly Sur Seine, FR)
Assignee:	Societe L\'Oreal S.A. (Paris, FR)
Application Number:	08/800,457
Patent Claims:	1. A method for testing whether a substance is suitable for use as a hair-care substance comprising the following steps: (i) isolating a fragment of at least one hair follicle which does not comprise dermal papilla by removing such hair follicle beneath the point of attachment of the sebaceous gland, while excluding the portion of the hair shaft located above the point of attachment of the sebaceous gland; (ii) placing the resulting hair follicle fragment in a suitable culture medium; (iii) incubating said hair follicle fragment with a substance which is to be screened for its ability to be used as a hair-care substance for a sufficient time to determine whether said substance affects expression of a marker substance wherein the said marker substance is type IV 72-kilodalton collagenase or type IV 92-kilodalton collagenase; and (iv) determining whether such substance is a suitable hair-care substance by quantifying the expression of said marker substance and comparing said value to marker substance expression by a control hair follicle fragment treated under identical conditions, except that it is not placed in contact with said substance which is being screened for its ability to unction as a suitable hair-care substance. 2. The method according to claim 1, wherein said marker substance is a substance the amount of which correlates to slowing or stopping of hair growth, hair loss, or the aggravated growth of hair follicles. 3. The method according to claim 1, wherein said hair follicle is isolated by dissection. 4. The method according to claim 1, wherein said follicle fragment exhibits at least one of the following morphological characteristics: (i) thickening of the basal membrane into a refringent eosinophilic layer which is located at the interface between the outer sheath of said hair follicle fragment and the connective sheath; (ii) a pattern of specific differentiation of the keratinocytes in the outer sheath, which differs from that of the keratinocytes in the upper part of the hair follicle, and from those comprised in the epidermis; (iii) lacks an antigen in the outer sheath which is recognized by an anti-desmoglein antibody; (iv) comprises in the outer sheath a basal lateral distribution of the integrins .alpha.2.beta.1 and .alpha.3.beta.1 in the basal keratinocyte layer; and (v) comprises an epidermal growth factor (EGF) receptor comprised on the surface of the basal keratinocytes and the suprabasal keratinocytes comprised in the outer sheath. 5. The method according to claim 4, wherein said hair follicle fragment exhibits all of said morphological features (I)-(V). 6. The method according to claim 1, wherein said hair follicle is in the anagen plase. 7. The method according to claim 1, wherein said culture medium is selected from the group consisting of Dulbecco MEM medium, Williams E medium, F12 medium, HAM medium, and RPMI1640 medium. 8. The method according to claim 7, wherein said culture medium is Williams E medium. 9. The method according to claim 1, wherein said hair follicle fragment is incubated for a time ranging between 10 seconds and 120 hours. 10. The method according to claim 9, wherein said incubation time ranges between 1 and 72 hours. 11. The method according to claim 1, wherein said marker which is quantified to assess the ability of said substance to function as a suitable hair-care substance is selected from the group consisting of a nucleic acid, a protein, a group of proteins, an ion, a cell organelle, a lipid, and a polyoside. 12. The method according to claim 1, wherein said marker which is quantified to assess the ability of said substance to function as a hair-care substance is quantified directly in the culture medium. 13. The method according to claim 1, wherein said marker, the expression of which is quantified to determine the ability of said screened substance to function as a hair-care substance, is measured in the cultured hair follicle fragment. 14. The method according to claim 13, wherein prior to quantitating the amount of said marker, the hair follicle fragment is ground. 15. A method for testing the ability of a particular substance to function as a suitable hair-care substance which comprises the following steps: (i) isolating a fragment of at least one hair follicle which does not comprise dermal papilla by removing said hair follicle at a point below the point of attachment of the sebaceous gland and excluding the portion of the hair shaft located above the point of attachment of the sebaceous gland; (ii) incubating said hair follicle fragment in a suitable culture medium in the presence of epidermal growth factor (EGF) or type .alpha. tumorous growth factor (TGF.alpha.) for a period of time (T1); (iii) placing said incubated hair follicle fragment in contact with a potentially active hair-care substance for a sufficient period of time to determine whether said potentially active hair-care substance has an effect on the expression of type IV 92-kilodalton collagenase; and (iv) quantitatively determining the amount of type IV 92-kilodalton collagenase and comparing the amount thereof to a control hair follicle fragment which is treated under identical conditions, except that said hair follicle fragment is not placed in contact with said potentially active hair-care substance. 16. The method according to claim 15, wherein said growth factor is contained in a concentration ranging from between 0.001 ng/ml and 100 ng/ml. 17. The method according to claim 16, wherein the concentration of said growth factor ranges between 0.01 ng/ml and 10 ng/ml. 18. The method according to claim 15, wherein said growth factor is epidermal growth factor (EGF). 19. The method according to claim 15, wherein said incubation time (T1) ranges between 5 seconds and 45 days. 20. The method according to claim 15, wherein said incubation time (T1) ranges between 1 hour and 72 hours. 21. The method according to claim 1, wherein said substance is screened for its ability to affect one of the following hair growth properties: (i) effect on slowing or retarding the growth of hair; (ii) affecting the loss of hair; and (iii) affecting the aggravated growth of hair follicles. 22. The method according to claim 1, wherein the ability of the substance to retard or prevent the growth and loss of hair follicles is determined. 23. The method according to claim 1, wherein said substance is screened for its ability to treat alopecia.

Details for Patent 5,891,426

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2016-12-15