Title:  Methods for altering hair growth and hair pigmentation by apoptosis in the follicular papillae and compositions therefor

United States Patent:  6,407,056

Assignee:  Johnson & Johnson Consumer Companies, Inc. (Skillman, NJ)

Filed:  June 25, 1997

The present invention utilizes serine proteases and their ability to induce programmed cell death and apoptosis in the follicular papillae to affect changes in mammalian hair growth and hair pigmentation. Also described are compositions which have an agent with a portion of similar structure to a portion of the trypsin molecule, allowing said agent to induce programmed cell death and apoptosis in the same manner as trypsin.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, "mammal" shall mean any member "of the higher vertebrae animals comprising the class Mammalia," as defined in Webster's Medical Desk Dictionary 407 (1986), and includes but is not limited to humans. As used herein "(%, w/v)" shall mean grams of a given component per 100 ml of the total composition. As used herein, "receptor" shall include both intracellullar and extracellular receptors and shall mean those molecules capable of receiving and transducing a signal.

Topically active agents suitable for use in the composition of the present invention include proteases, compounds having a structure substantially similar to a portion of the three-dimensional structure of trypsin, or mixtures thereof.

Preferred proteases are the serine proteases which include, but are not limited to, trypsin, carboxypeptidase-Y, protease IV, subtilysin, or mixtures thereof. The protease of choice is trypsin. While not willing to be bound by this theory, we believe that the proteases capable of delaying hair growth and hair pigmentation do so by altering the hair growth cycle via the induction of programmed cell death and apoptosis within the follicular papillae, and that this alteration is likely induced by a receptor-mediated signal transduction. Preferably, the protease is present in an amount, based upon the total volume of the composition of the present invention, of from about 0% (w/v) to about 5% (w/v), and more preferably from about 0.01% (w/v) to about 1% (w/v).

As evidenced by Example 10 herein, we have unexpectedly found that the proteolytic activity of a protease is not the only factor which contributes to the induction of programmed cell death. While not wishing to be bound by any theory, we further believe that a receptor-mediated signal transduction event may also contribute to the induction of programmed cell death of the follicular papillae, and that this type of agent might be related to a structural component of the topically active agent.

Therefore, a second suitable topically active agent includes compounds which possess a portion that is substantially similar in shape to the portion(s) of a three-dimensional structure of a trypsin molecule that affect this receptor-mediated signal transduction. As used herein, "substantially similar in shape" means that the compounds include a sufficient amount of a structure behaving like a ligand which either may occupy a receptor, may displace a ligand from the receptor, or may activate or inactivate a receptor by a proteolytic cleavage, and thereby contribute to the induction of programmed cell death. Examples of such compounds possessing portions which are substantially similar in shape to the structure of a portion of three-dimensional trypsin include, but are not limited to proteolytically inactive trypsin, which is a form of trypsin that is unable to digest proteins such as collagen: Preferably these compounds are present in the composition of the present invention in an amount, based upon the total volume of the composition, of from about 0% (w/v) to about 5% (w/v) and most preferably from about 0.01% (w/v) to about 1% (w/v).

If the delivery parameters of the topically active pharmaceutical or cosmetic agent so require, the topically active composition of the present invention may preferably be further comprised of a pharmaceutically or cosmetically acceptable vehicle capable of functioning as a delivery system to enable the penetration of the topically active agent into the hair follicle. While any commercially available vehicle for delivering the protease to the appropriate skin appendage, which in this case is the hair follicle, is suitable for use as the pharmaceutically or cosmetically acceptable vehicle, liposomes are preferred. The liposomes are more preferably non-ionic and comprised of a) glycerol dilaurate or glycerol distearate; b) compounds having the steroid backbone found in cholesterol; and c) fatty acid ethers having from about 12 to about 18 carbon atoms, wherein the constituent compounds of the liposomes are in a ratio of about 53:10:22 to about 63:20:32, and preferably from about 55:12:24 to about 61:18:30, respectively. Liposomes comprised of glycerol dilaurate/cholesterol/polyoxyethylene-10-stearyl ether (GDL) are most preferred. Preferably the liposomes are present in an amount, based upon the total volume of the composition, of from about 10 mg/mL to about 100 mg/mL, and more preferably from about 25 mg/mL to about 50 mg/mL. A ratio of about 58:15:27 is most preferred. Suitable liposomes may preferably be prepared in accordance with the protocol set forth in Example 2, though other methods commonly used in the art are also acceptable.

The above described composition may be prepared by combining the desired components in a suitable container and mixing-them under ambient conditions in any conventional high shear mixing means well known in the art for non-ionic liposomes preparations, such as those disclosed in Niemiec et al., "Influence of Nonionic Liposomal Composition On Topical Delivery of Peptide Drugs Into Pilosebacious Units: An In Vivo Study Using the Hamster Ear Model," 12 Pharm. Res. 1184-88 (1995) ("Niemiec"), which is incorporated by reference herein in its entirety.

In a preferred embodiment, the pH of the topically active pharmaceutical or cosmetic composition may be adjusted with a known pH adjuster including but not limited to N-2-hydroxyethylpiperazine-N'-2-ethane sulfonic acid available from, for example, Life Technologies under the tradename "Hepes", or (hydroxymethyl)aminomethane, available from, for example, Life Technologies under the tradename, "Tris", citric acid, and mixtures thereof to yield a pH of from about 2 to about 8 in the final composition.

In alternative embodiments, the topically active pharmaceutical or cosmetic composition may be optionally combined with other ingredients such as moisturizers, foaming agents, cosmetic adjuvants, anti-oxidants, surfactants, foaming agents, conditioners, humectants, fragrances, viscosifiers, buffering agents, preservatives, and the like in an amount which will not destroy the liposomal structure in order to produce cosmetic or pharmaceutical products such as, non-exclusively, shaving creams, shaving gels, shaving powders, chemical depilatory creams and the like.

In another embodiment of the present invention, we have found a process for affecting changes in mammalian hair growth and hair pigmentation comprised of applying the topically active pharmaceutical or cosmetic composition to the surface of an animal's skin at about the time of hair growth induction.

Hair growth induction from telogen follicles can be performed by any method well-known in the art, including, but not limited to, shaving, wax depilation, chemical depilation and combinations thereof.

Although the time at which the topically active pharmaceutical or cosmetic composition is applied to the skin as well as the amount of time for which the composition remains on the skin may vary, one skilled in the art would readily realize without undue experimentation that the topically active composition is preferably applied to the skin surface either immediately before, immediately after or simultaneously with hair growth induction from a telogen state. As used herein, "immediately" shall be defined as within a period of about one hour. More preferably, the topically active pharmaceutical or cosmetic composition is applied either simultaneously with or immediately following hair growth induction and is left on the skin for a period sufficient to delay hair growth, which is preferably at least about five minutes and more preferably is at least about fifteen minutes after the time of application.

The topically active pharmaceutical or cosmetic composition should be applied in an amount effective to affect changes in mammalian hair growth and hair pigmentation. As used herein "amount effective" shall mean an amount sufficient to cover the region of skin surface where a delay in hair growth and hair pigmentation is desired. Preferably, the composition is applied to the skin surface such that, based upon a square cm of skin surface, from about 2 .mu.l/cm² to about 8 .mu.l/cm² of topically active agent is present when a delay in hair growth and hair pigmentation is desired.

We have unexpectedly found that when topically active agents, such serine proteases, are topically applied to an animal's skin at about the time of hair growth induction, a significant delay in hair growth and hair pigmentation of at least about 9 days was achieved. We further believe that since the hair growth cycle for humans is often slower than that for mice, it is further likely that the hair growth delay in humans would be considerably longer than 9 days.

Claim 1 of 17 Claims

We claim:

1. A method for delaying mammalian hair growth comprising:

topically applying to a surface of the skin of a mammal an effective amount of a topically active composition comprising a serine protease and a pharmaceutically or cosmetically acceptable vehicle comprising at least one liposome.

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