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Title:  Use of a polypeptide for treatment of pruritus in animals

United States Patent:  6,969,590

Issued:  November 29, 2005

Inventors:  Lipton; James M. (Woodland Hills, CA); Catania; Anna P. (Milan, IT)

Assignee:  Zengen, Inc. (Woodland Hills, CA)

Appl. No.:  322577

Filed:  December 17, 2002

The present invention is directed to a treatment for animal pruritus. One aspect of this invention involves a treatment for animal pruritus comprising one or more polypeptides with an amino acid sequence including KPV (SEQ ID NO: 1), VKP-Ac-CC-Ac-KPV (SEQ ID NO: 5), MEHFRWG (SEQ ID NO: 2), HFRWGKPV (SEQ ID NO: 3) or SYSMEHFRWGKPV (SEQ ID NO: 4) for animal pruritus caused by exposure to any number of agents or causes. The polypeptides are at a level to effectively treat the animal pruritus and are combined with a shampoo. Other combinations include the polypeptides at a level to effectively treat animal pruritus combined with a shampoo and an antibiotic, antifungal and/or and anti-inflammatory. The one or more polypeptides can also be a dimer formed from any of the amino acid sequence above.

SUMMARY OF THE INVENTION

The invention includes a composition and method of treatment of animal pruritus. A preferred embodiment of the invention is composition for the treatment of animal pruritus comprising a therapeutically effective amount of one or more peptides having a COOH-terminal sequence consisting of KPV (SEQ ID NO: 1), MEHFRWG (SEQ ID NO: 2), HFRWGKPV (SEQ ID NO: 3), and SYSMEHFRWGKPV (SEQ ID NO: 4) in combination with a shampoo.

The one or more polypeptides may be a dimer formed from any of the amino acid sequences above. The dimer, VPK-Ac-CC-Ac-KPV (SEQ ID NO: 5) (Ac=Acetyl group), is N-acetylated and C-amindated. Dimers can be formed by adding cysteines at the N-termini of any of the above polypeptides and allowing the cysteines of two polypeptides to form a disulfide bond. Both homo-dimers and hetero-dimers can be formed using this method.

Another preferred embodiment of the invention is a composition for the treatment of animal pruritus comprising a therapeutically effective amount of one or more of these peptides in combination with a therapeutically effective amount of an anti-inflammatory and a shampoo.

In another embodiment of the invention each of these compositions may further comprise a therapeutically effective amount of an antibiotic.

In another embodiment of the invention each of these combination compositions may further comprise a therapeutically effective amount of an antifungal.

The peptides in each of these preferred combination compositions has the primary sequence of KPV (SEQ ID NO: 1) or VPK-Ac-CC-Ac-KPV (SEQ ID NO: 5). In a preferred composition, pharmacologically effective concentrations of the peptides may be as low as 10^-12 M but may be as high 10^-4 M.

Another embodiment of the invention is a method to treat animal pruritus comprising topical or systemic administration of one or more of the preferred peptides.

GENERAL DESCRIPTION OF THE INVENTION

The references cited above and below are incorporated by reference as if fully set forth herein. The present invention involves a composition and a method for treatment for animal pruritus utilizing α-MSH and/or its derivatives.

α-MSH is a 13 amino acid with the primary sequence SYSMEHFRWGKPV (SEQ ID NO: 4). In addition to anti-inflammatory properties, antibiotic and its anti-fungal properties, it also has anti-pyretic properties. The C-terminal tripeptide, KPV (SEQ ID NO: 1), appears responsible for these properties. Lipton, J. M., Antipyretic and Anti-inflammatory Lys-Pro-Val-Compositions and Methods of Use, U.S. Pat. No. 5,028,592, issued Jul. 2, 1991; Lipton, J. M., Antipyretic and Anti-inflammatory Lys-Pro-Val-Compositions and Methods of Use, U.S. Pat. No. 5,157,023, issued Oct. 20, 1992; Catania, A., Lipton J. M., α-Melanocyte Stimulating Hormone in the Modulation of Host Reactions, 14 Endocr. Rev., 564-576 (1993); Lipton, J. M., Catania, A., Anti-inflammatory Influence of the Neuroimmunomodulator α-MSH, 18 Immunol. Today, 140-145 (1997).

The core α-MSH sequence (4-10) has learning, memory and behavioral effects but limited anti-pyretic and anti-inflammatory activity. Lipton, J. M., Catania, A., Anti-inflammatory Influence of the Neuroimmunomodulator α-MSH, 18 Immunol. Today, 140-145 (1997). α-MSH, the α-MSH core and its tripeptide C-terminal have very low toxicity. Id.

α-MSH is produced by the post-translational processing of propriomelanocortin and shares the 1-13 primary sequence with adrenocortitrophic hormone (ACTH). Eberle, A. N., The Melanotropins, Karger, Basel, Switzerland (1988). It is secreted by a wide variety of cell types, including pituitary cells, monocytes, melanocytes, keratinocytes, epidermal cells and the epithelial cells of mucous membranes. Lipton, J. M., Catania, A., Anti-inflammatory Influence of the Neuroimmunomodulator α-MSH, 18 Immunol. Today, 140-145 (1997).

α-MSH reduces inflammation by modulating the inflammatory cascade locally and systemically. Rajora, N., Ceriani, G., Catania, A., Star, R. A., Murphy, M. T., Lipton, J. M., α-MSH Production, Receptors and Influence of Neopterin, in a Human Monocyte/macrophage Cell Line, 59 H. Leukoc. Biol., 248-253 (1996); Star, R. A., Rajora, N. Huang, J., Stock, R. C., Catania, A., Lipton, J. M., Evidence of Autocrine Modulation of Macrophage Nitric Oxide Synthase by α-MSH, 92 Proc. Natl. Acad. Sci., 8016-8020 (1995); Lipton, J. M., Ceriani, G., Macaluso, A., McCoy, D., Cames, K., Biltz, J., Catania, A., Anti-inflammatory Effects of the Neuropeptide α-MSH in Acute, Chronic and Systemic Inflammation, 741 Ann. N.Y. Acad. Sci., 137-148 (1994); Rajora, N., Boccoli, G., Burns, D., Sharma, S., Catania, A., Lipton, J. M., α-MSH Modulates Local Circulating Tumor Necrosis Factor A in Experimental Brain Inflammation, 17 J. Neurosci, 2181-2186 (1997); Richards, D. B., Lipton, J. M. Effect of α-MSH (11-13) (Lys-Pro-Val) on Fever in Rabbits, 5 Peptides, 815-817 (1984); Hiltz, M. E., Lipton, J. M., Anti-inflammatory Activity of a COOH-terminal Fragment of the Neuropeptide α-MSH, 3 FASEB J., 2282-2284 (1989).

α-MSH (1-13) derivatives are also effective in the treatment of animal pruritus. Derivatives include biologically functional equivalents and hydropathic amino acids, as described in Example I, infra, as well as selected amino acid sequences within the native α-MSH (1-13) chemical structure, i.e. KPV (SEQ ID NO: 1), MEHFRWG (SEQ ID NO: 2), HFRWGKPV (SEQ ID NO: 3) and VPK-Ac-C-C-Ac-KPV (SEQ ID NO: 5).

One aspect of the invention is a composition and method of treatment of animal pruritus having an inflammatory bacterial and/or fungal component. A preferred embodiment of the invention is a composition for the treatment of animal pruritus comprising a therapeutically effective amount of one or more peptides having a C-terminal sequence consisting of KPV (SEQ ID NO: 1), MEHFRWG (SEQ ID NO: 2), HFRWGKPV (SEQ ID NO: 3), SYSMEHFRWGKPV (SEQ ID NO: 4) and VPK-Ac-C-C-Ac-KPV (SEQ ID NO: 5) in combination with a shampoo.

Another preferred embodiment of the invention is a composition for the treatment of animal pruritus comprising a therapeutically effective amount of one or more peptides having a C-terminal sequence consisting of KPV (SEQ ID NO: 1), MEHFRWG (SEQ ID NO: 2), HFRWGKPV (SEQ ID NO: 3), SYSMEHFRWGKPV (SEQ ID NO: 4) and VPK-Ac-C-C-Ac-KPV (SEQ ID NO: 5) in combination with a therapeutically effective amount of a cortisol based glucocorticoid such as betamethasone, cortisone, dexamethasone, hydrocortisone, methylprednisone, prednisone, and triamcinolone and a shampoo.

Instead of glucocorticoids, a pharmacologically effective amount of non-steroidal anti-inflammatory drugs (NSAID) such as acetylsalicylic acid, diflusinal, fenoprophen calcium, ibuprofen, indomethacin, meclofenamate sodium, naproxen, phenylbutazone, piroxicam, sulindac, and tolmetin sodium may be employed in combination with the preferred peptides.

In another embodiment of the invention each of these compositions may comprise a therapeutically effective amount of an antibiotic such as quinalones, penicillins, lincomides, β-lactam inhibitors, cephalosporins, aminoglycocides, and tetracyclines.

In another embodiment of the invention each of these compositions may further comprise a therapeutically effective amount of an antifungal such as itraconazole, econazole, ketaconazole, miconazole and fluconazole.

More preferably still, the peptides in each of these preferred combination compositions has the primary sequence of KPV (SEQ ID NO: 1) or VPK-Ac-CC-Ac-KPV (SEQ ID NO: 5) (Ac=Acetyl group). In all the preferred compositions, pharmacologically effective concentrations of the peptides may be as low as 10^-12 M but may be as high 10^-4 M.

In yet another embodiment of the invention, one or one or more peptides having a C-terminal sequence of KPV, such as KPV (SEQ ID NO: 1), MEHFRWG (SEQ ID NO: 2), HFRWGKPV (SEQ ID NO: 3), SYSMEHFRWGKPV (SEQ ID NO: 4), and VPK-Ac-C-C-Ac-KPV (SEQ ID NO: 5) which may or may not be in combination with therapeutically effective amounts of antibiotics, corticosteroids, and/or antifungals is dissolved in a carrier. Formulations for solution or solid based drug delivery carriers are well known in the art. Such preferred carriers include, but are not limited to, saline, phosphate buffered saline, gelatin, maltodextrin, cellulose, microcrystalline cellulose, methyl cellulose and carboxymethyl cellulose.

The formulation of tablets are well known in the art. An exemplary formulation of a hard gelatinous tablet comprises:

An exemplary formulation of a hard tablet comprises:

Alternative carriers include common commercial formulations for shampoos, creams, ointments, balms, aerosol foams, gels or liquids. The carrier itself or a component dissolved in the carrier may have palliative or therapeutic properties of its own, including moisturizing, cleansing, or anti-inflammatory/anti-itching properties.

Formulations of creams and gels are well known in the art. HARRY'S COMSETICOLOGY (Chemical Publishing, 7^th ed. 1982); REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Publishing Co., 18^th ed. 1990).

By far, the most common cleansing agent used in the treatment of animals is a shampoo. Set forth below are examples of various formulations of the invention in different classifications of shampoos. Examples of some systemic preparations are also included showing the invention in those formulations. As used below the term "Active Ingredient" refers to one or more peptides having a C-terminal sequence of KPV, such as KPV (SEQ ID NO: 1), MEHRFWG, HRFWGKPV and SYSMEHFRWGKPV (SEQ ID NO: 4). Preferably, the active ingredient is KPV (SEQ ID NO: 1) or VPK-Ac-CC-Ac-KPV (SEQ ID NO: 5).

An exemplary formulation of a clear liquid shampoo based on the invention follows. (In these formulations, the designation of q.s. is meant to refer to a "quantity sufficient" for the desired effect.)

An exemplary formulation of a moisturizing shampoo based on the invention comprises:

Another exemplary formulation of a liquid cream or lotion shampoo based on the invention, comprises:

Another exemplary formulation or a solid cream or gel shampoo based on the invention comprises:

An exemplary formulation of an oil shampoo based on the invention comprises:
 

Certain pruritic conditions are caused by drainage from an exposed wound or injury. In such a case, a dry shampoo based on the invention would be preferable. An exemplary formulation of a dry shampoo based on the invention comprises:

 

Another preferred embodiment of the invention is a treatment packet with a wipe made of absorbent material that is treated with α-MSH and/or its derivatives that have been dissolved into a liquid-based carrier. This type of application would be most beneficial in those animals showing some level of alopecia (hair loss), or animals that have been intentionally shaved in localized areas of dermatologic involvement.

The process for making wipes of absorbent material is well known in the art. For example, baby wipes, moist towelettes, make-up removal cloths, and alcohol swabs are all wipes of absorbent material. Commercial examples of such wipes include Chubs® Baby Soft Wipes, Dexus® Antibacterial Hand Wipes, Dexus® Makeup Remover Wipes, Tinactin® Sports Wipes for Athlete's Foot, and B-D® Alcohol Swabs. Treatment of the wipe's absorbent material is accomplished by first soaking the absorbent material in a solution of α-MSH and/or its derivatives. The wipe remains in a liquid-impermeable packaging until use, when the package is opened and the wet wipe is applied to the affected portion of the integument.

The processes for making liquid-impermeable packages are well known in the art. For example, packages made of layers of paper, metal foil, and metal foil coated paper are commonly used for packaging wipes of absorbent material. For example, moist towelettes, such as Massengill® Feminine Cleansing Soft Cloth Towelettes, and alcohol swabs, such as B-D® Alcohol Swabs, are packaged in this manner.

In another embodiment of the invention, α-MSH, and/or its derivatives, may be administered parenterally. Preferred compositions for parenteral administration may be formed by combining the preferred peptides in combination with pharmaceutically acceptable buffers, dilutents and stabilizers. In a preferred composition, approximately 100-500 mg of the preferred peptides is mixed with about 1-7 ml of saline, including a pharmacologically acceptable buffer to maintain a neutral pH.

An exemplary parenteral preparation comprises:

Another preferred embodiment of the invention is a method for treating animal pruritus comprising systemic or topical application of a therapeutically effective level of α-MSH, one or more peptides with a C-terminal sequence of KPV such as KPV (SEQ ID NO: 1), and HFRWGKPV (SEQ ID NO: 3) or any sequence disclosed herein, including functionally equivalent derivatives. The peptides of this preferred method may be combined with a shampoo or therapeutically effective amounts of anti-inflammatories such as corticosteroids, fungicides, antibiotics, moisturizers or anti-itching compounds.

Topical administration preferably comprises direct topical application of a pharmacologically effective amount of one or more of the preferred peptides contained in a carrier to the affected regions. A preferred method comprises topical application of a shampoo, a moisturizer, or a moisturizing swab containing pharmacologically effective amounts of one or more of the preferred peptides. Alternatively, topical administration of a pharmacologically effective amount may utilize transdermal delivery systems well known in the art.

Systemic administration preferably comprises ingestion of any solid or solution carriers containing a pharmacologically effective amount of one or more of the preferred peptides. Such solid or solution carriers may comprise pills, hard tablets, soft tablets, gums or ordinary liquids. Additionally, systemic administration of a pharmacologically effective amount may comprise invasive methodologies including intravenous, subcutaneous, intramuscular or intralesional injection of a suitable carrier, such as saline, containing a pharmacologically effective amount of one or more of the preferred peptides.
 

Claim 1 of 13 Claims

1. A method comprising: the steps of:

selecting an animal; and

administrating a therapeutically effective amount of a polypeptide having a C-terminus sequence of KPV (SEQ ID NO: 1); and wherein the peptide is administered via a shampoo to treat animal pruritus.

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