WO2009120656A1

WO2009120656A1 - Pharmaceutical for ocular indications

Info

Publication number: WO2009120656A1
Application number: PCT/US2009/038024
Authority: WO
Inventors: John H. Dodd; Stephen Pitt
Original assignee: Palatin Technologies, Inc.
Priority date: 2008-03-24
Filing date: 2009-03-24
Publication date: 2009-10-01
Also published as: WO2009151714A3; WO2009151714A2

Abstract

Methods of treatment of dry eye and related ocular conditions and for prevention of corneal transplant rejection by administration of a therapeutically effective amount of a pharmaceutical preparation including a melanocortin agonist, and formulations and preparations for treatment of such conditions.

Description

Pharmaceutical for Ocular Indications

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to and the benefit of the filing of U.S. Provisional Patent Application Serial No. 61/038,800 entitled "Treatment of Circulatory Shock, Ischemia, Inflammatory Diseases and Related Conditions", filed on March 24, 2008, and the specification and claims thereof is incorporated herein by reference.

A related application entitled "Therapeutic for Treatment of Circulatory Shock, Ischemia, Inflammatory Disease and Related Conditions" is being filed concurrently herewith, U.S. Patent

Application , Attorney Docket No. 0903-094, and the specification and claims thereof are incorporated herein by reference.

BACKGROUND OF THE INVENTION Field of the Invention:

The present invention relates to compounds, compositions, formulations and methods for treatment of dry eye and related diseases, indications, conditions and syndromes, by administration of a pharmaceutical composition including a melanocortin receptor agonist such as Ac-Nle-cyc/o(-Asp-His-D- Phe-Arg-Trp-Lys)-OH. Description of Related Art:

Note that the following discussion refers to a number of publications by authors and year of publication, and that due to recent publication dates certain publications are not to be considered as prior art vis-a-vis the present invention. Discussion of such publications herein is given for more complete background and is not to be construed as an admission that such publications are prior art for patentability determination purposes.

Dry eye or dry eye syndrome is caused by either a deficiency in the quantity or the quality of the tears or tear film. It is a common eye condition which, according to some estimates, in the United States affects up to 1 1 % of people aged 30 to 60 years of age and 15% of those 65 years of age or older. The number of patients is likely to increase as the population ages, resulting in a decrease in the quality of life.

Dry eye, or keratoconjunctivitis sicca, results from any of a number of causes, including deficiencies in tear film layers and tear production, environmental factors such as exposure to air pollution or hot, dry, windy climates, activities such as prolonged computer use or intense reading, particularly when associated with a decrease in the frequency of blinking, effects resulting from contact lens, or as a side effect of medications or medical conditions, which medical conditions include hormonal changes such as associated with menopause, thyroid disease, Parkinson's disease, Sjorgren's syndrome and Vitamin A deficiency. Current therapeutic modalities include environmental or behavioral modifications, use of artificial tears, use of a cyclosporine ophthalmic emulsion such as Restasis® (Allergan), insertion of punctual plugs in the lacrimal puncta in the eyelids to decrease tear drainage, oral antibiotics such as tetracycline or use of anti-inflammatory eye drops such as corticosteroid eye drops. However, even with these therapeutic modalities there are a significant proportion of patients with less than optimal outcomes. It is against this background that the invention was made.

BRIEF SUMMARY OF THE INVENTION

This invention relates to methods for treatment of a mammalian patient with an ocular inflammatory disease, including administration to the eye of a patient with an ocular inflammatory disease of a therapeutically effective amount of a pharmaceutical composition including a melanocortin agonist. The melanocortin agonist may be a peptide melanocortin receptor agonist with binding at human MC1-R, as determined by Ki values, at least ten times as great as the binding at human MC4-R. The melanocortin agonist may also be a peptide melanocortin receptor agonist wherein the binding at human MC1-R, as determined by Ki values, is more than ten times as great as the binding at human MC4-R. In one aspect, the melanocortin agonist is a peptide melanocortin receptor agonist wherein the Ki at human MC1-R is less than 1.0 nM and the Ki at human MC4-R is greater than 1.0 nM but less than about 10 nM. In another aspect, the melanocortin agonist is the peptide Ac-Nle-cyc/o(-Asp-His-D-Phe- Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof. The ocular inflammatory disease may be uveitis or dry eye. Administration to the eye may include eye drops. The therapeutically effective amount may include between 1 and about 6 drops in each affected eye of an ophthalmic solution including between about 0.0000001 % and about 2% (w/v) of the peptide or pharmaceutically acceptable salt thereof, alternatively between about 0.000001 % and about 0.2% (w/v) of the peptide or pharmaceutically acceptable salt thereof, or alternatively between about 0.00001 % and about 0.2% (w/v) of the peptide or pharmaceutically acceptable salt thereof. Further provided is a solution adapted for ophthalmic administration, including between about 0.0000001 % and 2% (w/v) of a pharmaceutically acceptable salt of Ac-Nle-cyc/o(-Asp-His-D-Phe-Arg-Trp-Lys)-OH in an aqueous solution.

The invention further relates to providing a pharmaceutical composition including a peptide melanocortin receptor agonist wherein the binding at human MC 1 -R, as determined by Ki values, is at least ten times as great as the binding at human MC4-R.

The invention further relates to providing a pharmaceutical composition including a peptide melanocortin receptor agonist wherein the binding at human MC 1 -R, as determined by Ki values, is more than ten times as great as the binding at human MC4-R.

The invention further relates to providing a pharmaceutical composition including a peptide melanocortin receptor agonist wherein the Ki at human MC1-R is less than 1.0 nM and the Ki at human MC4-R is greater than 1.0 nM but less than 10 nM. -A-

The invention further relates to providing a pharmaceutical composition including a peptide melanocortin receptor agonist which is pharmaceutically effective through pathways involving agonism of both human MC1-R and human MC4-R.

Other aspects and novel features, and the further scope of applicability of the present invention will be set forth in part in the detailed description to follow, and in part will become apparent to those skilled in the art upon examination of the following, or may be learned by practice of the invention. The aspects of the invention may be realized and attained by means of the instrumentalities and combinations particularly pointed out in the appended claims.

DETAILED DESCRIPTION OF THE INVENTION

Definitions. Before proceeding with the description of the invention, certain terms are defined as set forth herein.

In the sequence given for bremelanotide according to the present invention, the amino acid residues have their conventional meaning as given in Chapter 2400 of the Manual of Patent Examining Procedure, 8^th Ed. Thus, "NIe" is norleucine, "Asp" is aspartic acid, "His" is histidine, "D-Phe" is D- phenylalanine, "Arg" is arginine, "Trp" is tryptophan, and "Lys" is lysine. "Ac" refers to a peptide or amino acid sequence that is acetylated.

The term "composition", as in pharmaceutical composition, is intended to encompass a product comprising the active ingredient(s), and the inert ingredient(s) that make up the carrier, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions utilized in the present invention encompass any composition made by admixing an active ingredient and one or more pharmaceutically acceptable carriers. In the context of the present invention, this includes any composition whereby an active ingredient may be introduced to any portion of the eye, including but not limited to the surface of the eye. A composition thus includes eye drops, either aqueous or otherwise, as well as emulsions, ointments, gels, ocular inserts, biodegradable ocular inserts, plugs liposomes, micro particles, nanoparticles, nanospheres or other devices or formulations adapted for or which may be employed to introduce at least one active ingredient to any portion of the eye, including but not limited to the surface of the eye. By a melanocortin receptor "agonist" is meant an endogenous substance, drug substance or compound, including a compound such as bremelanotide, which can interact with a melanocortin receptor and initiate a pharmacological response, including but not limited to adenyl cyclase expression, characteristic of the melanocortin receptor. For this invention, a melanocortin receptor agonist which is an agonist at melanocortin-4 receptor (MC4-R) and melanocortin-1 receptor (MC1-R) is preferred, and most preferred is a melanocortin receptor agonist which is an agonist at MC1-R, melanocortin-3 receptor (MC3-R), MC4-R and melanocortin-5 receptor (MC5-R).

By "inflammatory disease," also sometimes called an "inflammatory condition," is meant to include a disease or condition characterized in part by inflammatory mechanisms, such as specific T lymphocyte reactions or antibody-antigen interactions causing the recruitment of inflammatory cells and endogenous mediator chemicals, including but not limited to cytokines, which mediator chemicals include but are not limited to one or more of increased NF-κB activity, increased TNF-α production, increased IL-1 production and increased IL-6 production.

Clinical Applications. The compositions and methods disclosed herein can be used for both medical applications and animal husbandry or veterinary applications. Typically, the methods are used in humans, but may also be used in other mammals. The term "patient" is intended to denote a mammalian individual, and is so used throughout the specification and in the claims. The primary applications of this invention involve human patients, but this invention may be applied to laboratory, farm, zoo, wildlife, pet, sport or other animals.

Ocular Indications. Ocular inflammatory disease is a disease of the eye including dry eye disease, which affects approximately 10-20% of the population. This disease progressively affects larger percentages of the population as it ages, with the majority of these patients being women. In addition, almost everyone experiences ocular irritation, or the symptoms and/or signs of dry eye as a condition, from time to time under certain circumstances, such as prolonged visual tasking (e.g., working on a computer), being in a dry environment, using medications that result in ocular drying and so on. In individuals suffering from dry eye, the protective layer of tears that normally protects the ocular surface is compromised, a result of insufficient or unhealthy production of one or more tear components. This can lead to exposure of the surface of the eye, ultimately promoting desiccation and damage of surface cells. Signs and symptoms of dry eye include but are not limited to keratitis, conjunctival and corneal staining, redness, blurry vision, decreased tear film break-up time, decreased tear production, tear volume, and tear flow, increased conjunctival redness, excess debris in the tear film, ocular dryness, ocular grittiness, ocular burning, foreign body sensation in the eye, excess tearing, photophobia, ocular stinging, refractive impairment, ocular sensitivity, and ocular irritation. Patients may experience one or more of these symptoms. The excess tearing response may seem counterintuitive, but it is a natural reflex response to the irritation and foreign body sensation caused by the dry eye. Some patients may also experience ocular itching due to a combination of ocular allergy and dry eye symptoms. There are many possible variables that can influence a patient's signs or symptoms of dry eye including levels of circulating hormones, various autoimmune diseases (e.g. Sjogren's syndrome and systemic lupus erythematosus), ocular surgeries including PRK or LASIK, many medications, environmental conditions, visual tasking such as computer use, ocular fatigue, contact lens wear, and mechanical influences such as corneal sensitivity, partial lid closure, surface irregularities (e.g. pterygium), and lid irregularities (e.g. ptosis, entropion/ectropion, Pinguecula). Environments with low humidity, such as those that cause dehydration, can exacerbate or cause dry eye symptoms, such as sitting in a car with the defroster on or living in a dry climate zone. In addition, visual tasking can exacerbate symptoms. Tasks that can greatly influence symptoms include watching TV or using a computer for long periods of time where the blink rate is decreased. Uveitis is an ocular disease involving inflammation of the middle layer or uvea of the eye, and may also be understood to include any inflammatory process involving the interior of the eye. Uveitis includes anterior, intermediate, posterior and panuveitic forms, with the majority of uveitis cases anterior in location, involving inflammation of the iris and anterior chamber. This condition can occur as a single episode and subside with proper treatment or may take on a recurrent or chronic nature. Symptoms include red eye, injected conjunctiva, pain and decreased vision. Signs include dilated ciliary vessels, presence of cells and flare in the anterior chamber, and keratic precipitates on the posterior surface of the cornea. Intermediate uveitis includes inflammation and the presence of inflammatory cells in the vitreous cavity, and posterior uveitis include the inflammation of the retina and choroid. Uveitis may be secondary to any of a number of diseases and disorders, including acute posterior multifocal placoid pigment epitheliopathy, ankylosing spondylitis, Behget's disease, birdshot retinochoroidopathy, brucellosis, herpes simplex, herpes zoster, inflammatory bowel disease, juvenile rheumatoid arthritis, Kawasaki disease, leptospirosis, Lyme disease, multiple sclerosis, psoriatic arthritis, Reiter's syndrome, sarcoidosis, syphilis, systemic lupus erythematosus, toxocariasis, toxoplasmosis, tuberculosis, Vogt- Koyanagi-Harada syndrome, Whipple disease or polyarteritis nodosa.

Ocular inflammatory disease also includes inflammation and other conditions and syndromes associated with corneal transplant, also known as keratoplasty, in which a damaged cornea of a patient is replaced by the cornea from the eye of a donor, typically a human cadaver. Inflammation or other signs and symptoms of graft rejection occur in between about 5% to 30% of patients. Symptoms of inflammation or other graft rejection in corneal transplants include persistent discomfort, sensitivity to light, redness, or a change in vision. In one embodiment, ocular inflammatory disease, including but not limited to dry eye disease, uveitis, or inflammation or other conditions and syndromes associated with corneal transplant, may be treated with an ophthalmic dosage form including bremelanotide. The ophthalmic dosage form may include one or more active ingredients in addition to bremelanotide, such as for example artificial tear components, topical corticosteroids, non-steroidal anti-inflammatory drugs, or calcineurin inhibitors such as cyclosporine-A (Restasis® - Allergan). In a related embodiment, one or additional compounds may be given separately from bremelanotide, such as separate administration of an ophthalmic dosage form including an artificial tear component, a topical corticosteroid, a non-steroidal anti-inflammatory drugs, a calcineurin inhibitor such a cyclosporine-A, or a combination of any of the foregoing.

Methods of Administration and Use.

For ocular applications, in one aspect bremelanotide is formulated in an ophthalmic dosage form and is administered in the form of eye drops, eye washes or by means of other ocular delivery systems.

Compounds. In one aspect, the invention comprises a pharmaceutical composition including bremelanotide with the following general structure:

The peptide has a formula Of C₅₀H₆₈N₁₄Oi₀, and a net molecular weight of 1025.18. Bremelanotide may be synthesized by conventional means, including either solid-phase or liquid-phase techniques, and purified to greater than 99% purity, yielding a white powder that is a clear, colorless solution in water. Bremelanotide is an agonist at MC1-R, MC3-R, MC4-R and MC5-R. Utilizing NDP-α-MSH as a competitor, bremelanotide exhibits binding selectivity with a Ki value of approximately 0.2 nM at MC1-R, of approximately 9.0 nM at MC4-R, of approximately 72.0 nM at MC3-R and of approximately 159 nM at MC5-R. Bremelanotide has an EC₅₀ of approximately 3.0 nM at MC4-R under cell receptor densities approximating normal physiological systems with a relative efficacy, compared to NDP-α-MSH, in excess of 90%. Because bremelanotide is an extremely high potency molecule, it is expected that bremelanotide will be therapeutically effective at a concentration as low as 1.0 nM, very approximately corresponding to a solution of an ophthalmic formulation including 0.0000001 % (w/v) of bremelanotide or a pharmaceutically acceptable salt of bremelanotide, where the patient is to be administered between 1 and about 6 drops in the affected eye, or alternatively that bremelanotide will be therapeutically effective at a concentration as low as 10.0 nM, very approximately corresponding to a solution of an ophthalmic formulation including 0.000001 % (w/v) of bremelanotide or a pharmaceutically acceptable salt of bremelanotide, or alternatively that bremelanotide will be therapeutically effective at a concentration as low as 100 nM, very approximately corresponding to a solution of an ophthalmic formulation including 0.00001 % (w/v) of bremelanotide or a pharmaceutically acceptable salt of bremelanotide. Thus in one aspect a therapeutically effective amount of bremelanotide sufficient to treat ocular inflammatory disease is between about one and six drops of a solution containing between 0.0000001 % and about 2% (w/v) of bremelanotide or a pharmaceutically acceptable salt thereof, alternatively between about 0.000001 % and about 0.2% (w/v) of bremelanotide or a pharmaceutically acceptable salt thereof, or alternatively between about 0.00001 % and about 0.2% (w/v) of bremelanotide or a pharmaceutically acceptable salt thereof.

Peptide Synthesis. Bremelanotide may be readily synthesized by any known conventional procedure for the formation of a peptide linkage between amino acids. Such conventional procedures include, for example, any solution phase procedure permitting a condensation between the free alpha amino group of an amino acid or residue thereof having its carboxyl group or other reactive groups protected and the free primary carboxyl group of another amino acid or residue thereof having its amino group or other reactive groups protected. In a preferred conventional procedure, bremelanotide may be synthesized by solid-phase synthesis and purified according to methods known in the art. Any of a number of well-known procedures utilizing a variety of resins and reagents may be used to prepare bremelanotide. The process for synthesizing bremelanotide may be carried out by a procedure whereby each amino acid in the desired sequence is added one at a time in succession to another amino acid or residue thereof or by a procedure whereby peptide fragments with the desired amino acid sequence are first synthesized conventionally and then condensed to provide bremelanotide. The resulting peptide is then cyclized to yield a cyclic peptide of the invention.

Solid phase peptide synthesis methods are well known and practiced in the art. In such a method the synthesis of bremelanotide can be carried out by sequentially incorporating the desired amino acid residues one at a time into the growing peptide chain according to the general principles of solid phase methods. These methods are disclosed in numerous references, including, Merrifield, R. B., Solid phase synthesis (Nobel lecture). Angew Chem 24:799-810 (1985) and Barany et al., The

Peptides, Analysis, Synthesis and Biology, Vol. 2, Gross, E. and Meienhofer, J., Eds. Academic Press 1-284 (1980).

Reactive side chain groups of the various amino acid residues are protected with suitable protecting groups, which prevent a chemical reaction from occurring at that site until the protecting group is removed. Usually also common is the protection of the alpha amino group of an amino acid residue or fragment while that entity reacts at the carboxyl group, followed by the selective removal of the alpha amino protecting group to allow a subsequent reaction to take place at that site. Specific protecting groups have been disclosed and are known in solid phase synthesis methods and solution phase synthesis methods. Alpha amino groups may be protected by a suitable protecting group, including a urethane-type protecting group, such as benzyloxycarbonyl (Z) and substituted benzyloxycarbonyl, such as p- chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p-biphenyl- isopropoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc) and p-methoxybenzyloxycarbonyl (Moz); aliphatic urethane-type protecting groups, such as t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropoxycarbonyl, and allyloxycarbonyl. Fmoc is preferred for alpha amino protection. Guanidino groups may be protected by a suitable protecting group, such as nitro, p- toluenesulfonyl (Tos), Z, pentamethylchromanesulfonyl (Pmc), adamantyloxycarbonyl, and Boc. Pmc is a preferred protecting group for Arg.

Bremelanotide described herein was primarily prepared using solid phase synthesis. Solid phase synthesis was commenced from the C-terminal end of the peptide by coupling a protected alpha amino acid to a suitable resin. Such a starting material can be prepared by attaching an alpha amino- protected Lys amino acid by an ester linkage to a p-benzyloxybenzyl alcohol (Wang) resin or a 2- chlorotrityl chloride resin, by an amide bond between an Fmoc-Linker, such as p-[(R, S)-α-[1-(9H-fluor- en-9-yl)-methoxyformamido]-2,4-dimethyloxybenzyl]-phenoxyacetic acid (Rink linker) to a benzhydrylamine (BHA) resin, or by other means well known in the art. Fmoc-Linker-BHA resin supports are commercially available and generally used when feasible. The resins are carried through repetitive cycles as necessary to add amino acids sequentially. The alpha amino Fmoc protecting groups are removed under basic conditions. Piperidine, piperazine, diethylamine, or morpholine (20- 40% v/v) in DMF may be used for this purpose. Following removal of the alpha amino protecting group, the subsequent protected amino acids are coupled stepwise in the desired order to obtain an intermediate, protected peptide-resin. The activating reagents used for coupling of the amino acids in the solid phase synthesis of peptides are well known in the art. After bremelanotide is synthesized, if desired, the orthogonally protected side chain protecting groups may be removed using methods well known in the art for further derivatization of the peptide.

Reactive groups in a peptide can be selectively modified, either during solid phase synthesis or after removal from the resin. For example, the N-terminus acetylation may be accomplished while the peptides on resin, or the peptides may be removed from the resin by use of a cleaving reagent and then modified. Methods for N-terminus modification, such as acetylation are well known in the art. Bremelanotide can, in one embodiment, be cyclized prior to cleavage from the peptide resin.

For cyclization through reactive side chain moieties, the desired side chains are deprotected, and the peptide suspended in a suitable solvent and a cyclic coupling agent added. Suitable solvents include, for example DMF, dichloromethane (DCM) or 1-methyl-2-pyrrolidone (NMP). Suitable cyclic coupling reagents include, for example, 2-(1 H-benzotriazol-1-yl)-1 ,1 ,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(7-aza-1 H-benzotriazol-1-yl)-1 ,1 ,3,3-tetramethyluronium tetrafluoroborate (TATU), 2-(2-oxo- 1(2H)-pyridyl)-1 ,1 ,3,3-tetramethyluronium tetrafluoroborate (TBTU) or N.N'-dicyclohexylcarbodiimide/i- hydroxybenzotriazole (DCCI/HOBt). Coupling is conventionally initiated by use of a suitable base, such as N,N-diispropylethylamine (DIPEA), sym-collidine or N-methylmorpholine (NMM).

Following cleavage bremelanotide from the solid phase following synthesis, bremelanotide can be purified by any number of methods, such as reverse phase high performance liquid chromatography (RP-HPLC), using a suitable column, such as a Ci₈ column. Other methods of separation or purification, such as methods based on the size or charge of the peptide, can also be employed. Once purified, bremelanotide can be characterized by any number of methods, such as high performance liquid chromatograph (HPLC), amino acid analysis, mass spectrometry, and the like.

Salt Forms. Bremelanotide may be in the form of any pharmaceutically acceptable salt. The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable nontoxic bases or acids including inorganic or organic bases and inorganic or organic acids. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, lithium, magnesium, potassium, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl- morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like. An acid addition salt, prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids, is particularly preferred. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, carboxylic, citric, ethanesulfonic, formic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, malonic, mucic, nitric, pamoic, pantothenic, phosphoric, propionic, succinic, sulfuric, tartaric, p-toluenesulfonic acid, trifluoroacetic acid, and the like. Acid addition salts of bremelanotide are prepared in a suitable solvent and an excess of an acid, such as hydrochloric, hydrobromic, sulfuric, phosphoric, acetic, trifluoroacetic, citric, tartaric, maleic, succinic or methanesulfonic acid. In one preferred embodiment, bremelanotide is in an acetate salt form, and is formulated in an aqueous solution, optionally including one or more salts, such as sodium chloride.

Ocular Administration. Bremelanotide or other melanocortin receptor agonists are administered in a form and by a method resulting in delivery of bremelanotide or other melanocortin receptor agonists to the eye or to a portion or part of the eye. In one aspect for the treatment of diseases of the eye, ophthalmic preparations for ocular administration including between about 0.0000001 % and about 5% by weight solutions or suspensions of a melanocortin receptor agonist in an acceptable ophthalmic formulation may be used. Emulsions, ointments, gels, ocular inserts, biodegradable ocular inserts, liposomes, microparticles, nanoparticles, nanospheres or ion pairing formulations may also be employed, which may, in some instances, result in increasing the ocular residence times of bremelanotide or other melanocortin receptor agonists. In one embodiment, the ophthalmic formulation is a solution that includes between about 0.0000001 % and about 5% (w/v) of bremelanotide or a salt thereof, alternatively between about 0.000001 % and about 0.2% (w/v) of bremelanotide or a salt thereof, or alternatively between about 0.00001 % and about 0.2% (w/v) of bremelanotide or a salt thereof.

In one aspect, an ophthalmic solution is employed, for direct administration to the eye, such as by way of eye drops. The ophthalmic solutions are preferably maintained in a pH range between about pH 3.5 to 9.0, and preferably about pH 6.5 and pH 7.2, with a suitable buffer. The pH may be adjusted by any known means, such as by use of HCI or NaOH. Buffers may include acetate, boric acid, sodium borate, potassium citrate, citric acid, sodium bicarbonate, TRIS, various mixed phosphate buffers (such as combinations of Na₂HPO₄, NaH₂PO₄ and KH₂PO₄) and mixtures thereof. Generally, buffers will be used in amounts ranging from about 0.05% to 2.5% (w/v), and preferably from about 0.1 % to 1.5% percent; buffers should be as close to physiological ion concentrations as possible to minimize potential irritation but still maintain drug product pH over the shelf life of the product.

The ophthalmic solutions employed in the present invention may be made from purified water, and in one aspect preferably from a physiological saline solution. Additional tonicity enhancing agents may be employed, including either ionic or non-ionic tonicity enhancing agents, or both. Ionic tonicity enhancing agents include alkali metal or earth metal halides, such as CaCI₂, KBr, KCI, LiCI, NaI, NaBr, NaCI, Na₂SO₄ or boric acid. Non-ionic tonicity enhancing agents include urea, glycerol, sorbitol, mannitol, propylene glycol, or dextrose. The aqueous solutions of the present invention are typically adjusted with tonicity agents to approximate an osmotic pressure equivalent to a 0.9% (w/v) solution of sodium chloride or a 2.5% solution of glycerol. However, tonicity ranges equivalent to between 0.7% and 1.5% NaCI are generally considered to be acceptable.

The solutions can also contain conventional, pharmaceutically acceptable preservatives, stabilizers, cosolvents and/or penetration enhancers as well as viscoelastic substances included in artificial tear preparations. Pharmaceutically acceptable preservatives include quaternary ammonium compounds such as benzalkonium chloride, benzoxonium chloride or the like; alkyl-mercury salts of thiosalicylic acid such as thiomersal, phenylmercuric nitrate, phenylmercuric acetate or phenylmercuric borate; sodium perborate; sodium chlorite; parabens, such asmethylparaben or propylparaben; alcohols such as chlorobutanol, benzyl alcohol or phenyl ethanol; guanidine derivatives such as chlorohexidine or polyhexamethylene biguanide; sorbic acid; boric acid; or peroxide forming preservatives, or combinations of two or more of the foregoing. Pharmaceutically acceptable antioxidants and chelating agents may be used including various sulphites (such as sodium metabisulphite, sodium thiosulphate, sodium bisulfite, or sodium sulfite), α-tocopherol, ascorbic acid, acetylcysteine, 8-hydroxyquinolome, antipyrine, butylated hydroxyanisole or butylated hydroxytoluene, EDTA, and others. Cosolvents such as alcohols and others may also be used. Various substances can also be used to enhance formulation stability, such as cyclodextrins.

Penetration enhancers may be employed in ophthalmic solutions, including compounds such as surfactants, certain organic solvents such as dimethylsulphoxide and other sulphoxides, dimethylacetamide and pyrrolidine, certain amides of heterocyclic amines, glycols (e.g. propylene glycol), propylene carbonate, oleic acid, alkylamines and derivatives, various cationic, anionic and nonionic surfactants, amphoteric surfactants and the like. Additional penetration enhancers that may be employed include cetylpyridinium chloride, ionophores such as lasalocid, benzalkonium chloride, polysorbates such as polysorbate 20 (Tween® 20), parabens, saponins, various polyoxyethylene ether compounds such as Brij® 35, Brij® 78 or Brij® 98, ethylenediaminetetraacetic acid (EDTA), bile salts, and bile acids (such as sodium cholate, sodium taurocholate, sodium glycodeoxycholate, sodium taurodeoxycholate, taurocholic acid, chenodeoxycholic acid and ursodeoxycholic acid), capric acid, azone, fucidic acid, hexamethylene lauramide, saponins, hexamethylene octanamide, and decylmethyl sulfoxide. Ion pairing formulations utilizing charged excipients or counter ions to shield/neutralize charged groups on drug molecules may also be employed to lower the lipophilicity of the compound to increase corneal penetration. These formulations include but are not limited to ions such as sorbic acid, boric acid and maleic acid, among other charged ion pairing agents.

Viscosity enhancers or lubricants may be employed as necessary or appropriate. In one aspect, the viscosity enhancer includes a water soluble polymer, such as polyols, including polyvinyl alcohol, a polyethylene glycol, or combinations of water soluble polymers. In one aspect, polyethylene glycol 300 or 400 is employed. The contents of water soluble polymer may be between about 0.25% and about 4.0% (w/v). Thus an ophthalmic solution can include, by way of example, 1 % of polyvinyl alcohol, 1 % of polyethylene glycol 300 or 400, or both. Other polyols may be employed, including glycerol, glycerin, polysorbate 80, propylene glycol, ethylene glycol, povidone, and polyvinylpyrrolidone. Other lubricants, sometimes also called tear substitutes, may also be employed, including cellulose derivatives such hydroxy propyl methyl cellulose, carboxymethyl cellulose sodium, hydroxypropyl cellulose, hydroxyethyl cellulose, and methyl cellulose; dextrans such as dextran 70; water soluble proteins such as gelatin; carbomers such as carbomer 934P, carbomer 941 , carbomer 940 and carbomer 974P; and gums such as HP-guar, xanthan gum or combinations thereof. Other viscosity enhancers that can be employed include polysaccharide compounds, such as sulfated or non-sulfated glycosaminoglycan compounds. In one aspect, the polysaccharide compound is a non-sulfated glycosaminoglycan such as hyaluronic acid or a pharmaceutically acceptable salt thereof, such as sodium hyaluronate. Any commercially available molecular weight range of hyaluronic acid or salts thereof may be employed. From about 0.05% to about 0.4% (w/v) of hyaluronic acid or a salt thereof may be employed in an ophthalmic solution. In another aspect, the polysaccharide compound is a non-sulfated glycosaminoglycan such as dextran. In yet another aspect, the polysaccharide is a sulfated glycosaminoglycan such as chondroitin sulfate.

Semi-solid formulations may be employed for ophthalmic delivery to increase corneal residence times of drug molecules. Ointments containing polyethylene glycols, lanolin alcohols, ozokerite, ceresin, microcrystalline wax, surfactants, preservatives, sorbitan monolaurate, white petrolatum and light liquid petrolatum (mineral oil) or other petrolatum like bases may be used. Aqueous or non-aqueous suspensions may also be used. For hydrophilic peptides, suspensions using pharmaceutically acceptable oils or petrolatum may be used. Suspensions may contain microspheres or microparticulates, nanoparticulates, mucoadhesive particles, viscosity increasing agents, surfactants and other agents. Mucoadhesive compounds include synthetic polymers, such as polyacrylic acid and polycarbophil; biopolymers such as hyaluronic acid or sodium carboxy methylcellulose (CMC); polyanionic polymers such as polyacrylic acid (PAA); polyacrylic acids such as Carbopol® 934P, polycarbophil, and CMC or PAA with Pluronic® polyoxalkylene ethers; or polycationic polymers such as chitosan. Emulsions (oil in water or water in oil), including microemulsions, may also be employed that are composed of pharmaceutically acceptable oils together with one or more of viscosity increasing agents, preservatives, cosolvents, surfactants and other agents. Pharmaceutically acceptable oils include mineral oils and organic oils, including oils comprising medium chain or long chain saturated or unsaturated fatty acids or esters thereof. Pharmaceutically acceptable oils thus include any of a range of medium chain triglycerides, as well as oils such as almond oil, castor oil, cottonseed oil, glycerin (glycerol), peanut oil, mineral oil, polyethylene glycol, poppyseed oil, propylene glycol, safflower oil, sesame oil, soybean oil, olive oil and vegetable oil. A surfactant such as a polyoxyethylene alkyl ether, polyoxyl castor oil, tyloxapol, alkyl aryl ether sulfonate, lecithin, sorbitan esters, glyceryl monostearate, cetyl alcohol, octoxynol-9, nonoxynol-9, polyoxyethylene stearates, polyoxyethylene sorbitan fatty acid esters such as polysorbate 20, 60 and 80 or others may also be employed. Aqueous gels, often comprised of polymers such as polyvinyl alcohol (PVA), polyacrylamide, poloxamer, hydroxypropyl methylcellulose (HPMC), carbomer, polymethylvinylether maleic anhydride, and hydroxypropyl ethylcellulose may also be used. Hydrogels containing swellable, water insoluble polymers may be utilized containing polymers such as poly(acrylic acid), poly(acrylic acids), poly(acrylamide), and ethylene maleic anhydride, and chemically or thermally-treated gelatins. Ocular inserts, liposomes, discomes, niosomes, dedrimers, nanosuspensions, nanoparticles and microparticles may also be used to provide a controlled release of the drug. Liposomes and other controlled release agents may be positively charged to increase residence times through ionic interactions with the negatively charged corneal surface. Nanoparticles may be composed of biodegradable polymers such as polyactides (PLAs), polycyano acrylates, poly (D,L-lactides), and natural polymers such as chitosan, gelatin, sodium alginate, albumin and others. Combination ophthalmic solutions may be employed, including specifically solutions including more than one active pharmaceutical ingredient. In one aspect, a non-steroidal anti-inflammatory drug (NSAID) is employed in combination with a melanocortin receptor agonist, including but not limited to bremelanotide. NSAIDs suitable for use in combination ophthalmic solutions include agents, their esters and pharmaceutically acceptable salts thereof that inhibit the cycloxygenase (COX)-I and/or -2 enzyme, including but not limited to propionic acid compounds such as naproxen, flurbiprofen, oxaprozin, ibuprofen, ketoprofen, fenoprofen; ketorolac tromethamine; acetic acid derivatives such as sulindac, indomethacin, and etodolac; phenylacetic acids such as diclofenac, bromfenac, and suprofen; arylacetic prodrugs such as nepafenac, and amfenac; salicyclic acids, such as aspirin, salsalate, diflunisal, choline magnesium trisalicylate; para-aminophenol derivatives such as acetaminophen; naphthylalkanones such as nabumetone; enolic acid derivatives such as piroxicam and meloxicam; femanates such as mefenamic acid, meclofenamate and flufenamic acid; pyrroleacetic acids such as tolmetin; and pyrazolones such as phenylbutazone; and COX-2 selective inhibitors such as celecoxib, valdecoxib, parecoxib, etoricoxib, and luaricoxib. The ophthalmic solutions may additionally comprise other active ingredients, including, but not limited to, vasoconstrictors, anti-allergenic agents, anti- infectives, steroids, anesthetics, anti-inflammatories, analgesics, dry eye treatment agents (e.g. secretagogues, mucomimetics, polymers, lipids, antioxidants), and the like, or be administered in conjunction (simultaneously or sequentially) with pharmaceutical compositions comprising other active ingredients, including, but not limited to, vasoconstrictors, anti-allergenic agents, anti-infectives, steroids, anesthetics, anti-inflammatories, analgesics, dry eye treatment agents (e.g. secretagogues, mucomimetics, polymers, lipids, antioxidants), and the like. Without limiting the generality of the foregoing, the following formulations (w/v unless otherwise specified) are intended to be exemplary, and to provide aqueous formulations suitable for administration to the eye by a patient, such administration to be on a schedule determined by a prescribing physician, such as between about one and four time daily.

First formulation:

Bremelanotide 0.0000001 % to 2%

Phosphate buffer 10 mM

NaCI 0.9%

Benzalkonium chloride 0.01 %

Second formulation: Bremelanotide 0.0000001 % to 2%

Boric acid 0.5%

Hydroxypropyl methylcellulose (HPMC) 0.15%

Edetate disodium (dihydrate) 0.1 %

Third formulation:

Bremelanotide 0.00000001 % to 5%

Sorbic acid 0.09%

Edetate disodium (dihydrate) 0.01 %

Benzalkonium chloride 0.01 %

Glycerin 2.5%

Fourth formulation:

Bremelanotide 0.0000001 % to 2%

Boric acid 1.40%

Edetate disodium (dihydrate) 0.01 %

Benzalkonium chloride 0.01 %

NaCI 0.2%

Fifth formulation:

Bremelanotide 0.00000001 % to 2%

Maleic acid 0.05%

Edetate disodium (dihydrate) 0.01 %

Chlorobutanol 0.5%

Glycerin 2.5% Sixth formulation:

Bremelanotide 0.0000001 % to 2% Castor oil 5.0% Tween 80 4.0% Glycerin 2.0% Carbomer 1342 0.05%

Therapeutically Effective Amount. In general, the actual quantity of bremelanotide administered to a patient will vary between fairly wide ranges depending upon the mode of administration, the formulation used, the specific ocular inflammatory disease and the response desired. The dosage for treatment is administration, by any of the foregoing means or any other means known in the art, of an amount sufficient to bring about the desired therapeutic effect. Thus in one aspect a therapeutically effective amount includes an amount of bremelanotide that is sufficient to therapeutically alleviate dry eye or other ocular diseases or disorders. In another aspect a therapeutically effective amount includes an amount of bremelanotide that is sufficient to limit or ameliorate rejection of transplanted corneas. The response to the therapeutically effective amount may be a cellular, organ or tissue-specific response, or a system response.

Alternative Melanocortin Receptor Agonists. While certain embodiments of the present invention are described primarily in the context of bremelanotide, it is to be understood that other melanocortin receptor agonists may be employed in some aspects. For example, the metallopeptide melanocortin receptor agonists disclosed in WO 02/064091 , filed on February 13, 2001 , and U.S. Patent No. 7,307,063, issued on December 11 , 2007, both entitled Melanocortin Metallopeptides for Treatment of Sexual Dysfunction; and WO 01/13112, filed on June 14, 2000, entitled Melanocortin Metallopeptide Constructs, Combinatorial Libraries and Applications, may be employed. In addition, the peptidomimetic melanocortin receptor agonists disclosed in U.S. Serial No. 10/776,419, filed on February 10, 2004, entitled Peptidomimetics of Biologically Active Metallopeptides; the pyrrolidine melanocortin receptor agonists disclosed in U.S. Patent No. 7,189,755, issued on March 13, 2007, entitled Pyrrolidine Melanocortin-Specific Compounds; and the bicyclic melanocortin receptor agonists disclosed in WO 05/017574, filed on January 20, 2004, entitled Bicyclic Melanocortin-Specific Compounds, may also be employed. Also particular preferred are the piperazine melanocortin agonists disclosed in WO 05/102340, filed on January 20, 2004 and U.S. Patent No. 7,354,923, issued on April 8, 2008, both entitled Piperazine Melanocortin-Specific Compounds; the melanocortin agonists disclosed in WO 03/006620, filed on July 11 , 2002, entitled Linear and Cyclic Melanocortin Receptor- Specific Peptides; WO 04/005324, filed on July 9, 2003, entitled Peptide Compositions for Treatment of Sexual Dysfunction; WO 01/00224, filed on June 29, 2000 and U.S. Patent No. 6,794,489, issued on September 21 , 2004, entitled Compositions and Methods for Treatment of Sexual Dysfunction; and U.S. Patent No. 7,176,279, issued on February 13, 2007, entitled Cyclic Peptide Compositions and Methods for Treatment of Sexual Dysfunction. The entire disclosure of each of the foregoing are incorporated here by reference.

It is to be understood that the foregoing listing of patent applications disclosing melanocortin receptor agonists is intended to only be exemplary, and that other melanocortin receptor agonists, whether heretofore known or hereafter developed, may similarly be used in the practice of aspects of this invention. It is to be understood that the contents of the above published patent applications, and in particular the general formulas and exemplified compounds therein, are incorporated in this specification in their entirety by reference thereto. It is further to be understood that the foregoing listing is intended to only be exemplary, and that other melanocortin receptor agonists, whether heretofore known or hereafter developed, may similarly be used in the practice of this invention.

Although the invention has been described in detail with particular reference to these preferred embodiments, other embodiments can achieve the same results. Variations and modifications of the present invention will be obvious to those skilled in the art and it is intended to cover all such modifications and equivalents. The entire disclosures of all references, applications, patents, and publications cited above are hereby incorporated by reference.

Claims

CLAIMS What is claimed is:

1. A method for treatment of a mammalian subject with an ocular inflammatory disease, comprising administration to an eye of the subject with an ocular inflammatory disease of a therapeutically effective amount of a pharmaceutical composition comprising a melanocortin agonist.

2. The method of claim 1 wherein the melanocortin agonist is a peptide melanocortin receptor agonist with binding at human MC1-R, as determined by Ki values, at least ten times as great as the binding at human MC4-R.

3. The method of claim 1 wherein the melanocortin agonist is a peptide melanocortin receptor agonist wherein the binding at human MC1-R, as determined by Ki values, is more than ten times as great as the binding at human MC4-R.

4. The method of claim 1 wherein the melanocortin agonist is a peptide melanocortin receptor agonist wherein the Ki at human MC1-R is less than 1.0 nM and the Ki at human MC4-R is greater than 1.0 nM but less than about 10 nM.

5. The method of claim 1 wherein the melanocortin agonist is a peptide melanocortin receptor agonist with a C-terminal free acid group.

6. The method of claim 1 wherein the melanocortin agonist is the peptide Ac-Nle-cyc/o(- Asp-His-D-Phe-Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof.

7. The method of claim 1 , wherein the ocular inflammatory disease is uveitis or dry eye.

8. The method of claim 1 , wherein the ocular inflammatory disease is associated with a corneal transplant or corneal transplant rejection.

9. The method of claim 1 , wherein the pharmaceutical composition comprises eye drops.

10. The method of claim 6, wherein the therapeutically effective amount comprises between 1 and about 6 drops in each affected eye of an ophthalmic solution comprising between about 0.0000001 % and 2% (w/v) of Ac-Nle-cyc/o(-Asp-His-D-Phe-Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof.

11. The method of claim 6, wherein the therapeutically effective amount comprises between 1 and about 6 drops in each affected eye of an ophthalmic solution comprising between about 0.000001 % and 0.2% (w/v) of Ac-Nle-cyc/o(-Asp-His-D-Phe-Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof.

12. A solution adapted for ophthalmic administration, comprising between about 0.0000001 % and 2% (w/v) of the peptide Ac-Nle-cyc/o(-Asp-His-D-Phe-Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof in an aqueous ophthalmic solution.

13. The solution of claim 12, comprising between about 0.000001 % and 0.2% (w/v) of the peptide Ac-Nle-cyc/o(-Asp-His-D-Phe-Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof.

14. The solution of claim 12, further comprising a pharmaceutically acceptable oil.

15. The solution of claim 14 wherein the pharmaceutically acceptable oil comprises almond oil, castor oil, cottonseed oil, glycerin (glycerol), peanut oil, mineral oil, polyethylene glycol, poppyseed oil, propylene glycol, safflower oil, sesame oil, soybean oil, olive oil or vegetable oil.

16. The solution of claim 12, further comprising a penetration enhancer.

17. The solution of claim 12, further comprising a viscosity enhancer or lubricant.

18. The solution of claim 12, further comprising a second active pharmaceutical ingredient.

19. Use of the peptide Ac-Nle-cyc/o(-Asp-His-D-Phe-Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof for the manufacture of a ophthalmic medicament for use in the treatment of a patient with an ocular inflammatory disease.

20. The use of claim 19 wherein the ocular inflammatory disease is dry eye disease or uveitis.

21. Use of the peptide Ac-Nle-cyc/o(-Asp-His-D-Phe-Arg-Trp-Lys)-OH or a pharmaceutically acceptable salt thereof for the manufacture of a ophthalmic medicament for use in the treatment of inflammation associated with corneal transplant or corneal transplant rejection in a patient with a corneal transplant.