Diamine-linked receptor-specific cyclic peptides

Abstract

Melanocortin receptor-specific peptides for use in pharmaceutical applications, particularly in the treatment of inflammation-related diseases, indications, conditions and syndromes, are provided. [Solution] Formula (II) JPEG2026041738000090.jpg60170 or a pharmaceutically acceptable salt of any of the foregoing.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is related to U.S. Provisional Patent Application No. 62 / 969,311, entitled "Diamine-Linked Receptor-Specific Cyclic Peptides," filed February 3, 2020, and U.S. Provisional Patent Application No. 63 / 124,927, entitled "Diamine-Linked Receptor-Specific Cyclic Peptides," filed December 14, 2020, the specifications and claims of each of the foregoing provisional patent applications are incorporated herein by reference.

[0002] Background of the Invention Field of invention (field of technology): The present invention relates to side chain-to-tail diamine-linked cyclic peptides, including receptor-specific peptides that are agonists, partial agonists, antagonists, or mixed agonist-antagonists at melanocortin receptors, and the use of melanocortin receptor-specific diamine-linked cyclic peptides in the treatment of melanocortin receptor-mediated diseases, indications, conditions, and syndromes. [Background technology]

[0003] Description of related technology: Peptides are cyclized through the side chains of two amino acid residues, one proximal to the N-terminus and the other proximal to the C-terminus of the peptide sequence, typically through a disulfide bond (e.g., through the side chains of two Cys residues) or through an amide bond (e.g., through the side chains of two residues, one having a side chain containing a carboxyl group and one having a side chain containing an amine). Head-to-tail cyclized peptides are also known, such as peptides in which an amide is formed by coupling an N-terminal group (e.g., an amine) and a C-terminal group (e.g., a carboxylic acid), thereby forming an amide-linked cyclic peptide.

[0004] A family of melanocortin receptor types and subtypes has been identified, including melanocortin receptor-1 (MC1r), which is expressed on normal human melanocytes, melanoma cells, macrophages, and other cells; melanocortin receptor-2 (MC2r) for ACTH (adrenocorticotropic hormone), which is expressed in cells of the adrenal gland; melanocortin receptor-3 and melanocortin receptor-4 (MC3r and MC4r), which are expressed in cells of the hypothalamus, midbrain, brainstem, and peripheral tissues; and melanocortin receptor-5 (MC5r), which is expressed in a wide distribution of peripheral tissues. MC1r is thought to be involved in inflammation, mediation of hair and skin pigmentation, and other functions; MC2r is thought to mediate steroid production; MC3r is thought to be involved in energy homeostasis, feeding behavior, mediation of inflammation, and other functions; MC4r is thought to be involved in feeding behavior, energy homeostasis, sexual function, and other functions; and MC5r is thought to be involved in regulating the exocrine system and other functions.

[0005] Both agonist and antagonist melanocortin receptor-specific compounds are known, including agonist and antagonist peptides.For example, MC4r agonist peptides are believed to be useful for treating obesity or inducing weight loss and for treating various forms of sexual dysfunction, including male erectile dysfunction and female sexual dysfunction.MC4r antagonist peptides are believed to result in weight gain, and are potentially useful for conditions such as cachexia and other wasting syndromes and conditions.

[0006] Peptide of the endogenous agonist, alpha-melanocortin-stimulating hormone (α-MSH) Analogs are known. These include both linear and cyclic peptides. Cyclic melanocortin receptor-specific peptides are generally cyclized through a side chain such as an amide or cysteine ​​linkage, acylated at the N-terminus, and amidated at the C-terminus (endogenous α-MSH is acylated at the N-terminus and amidated at the C-terminus), although it is known that α-MSH analogs can have a C-terminal carboxyl group, as disclosed in U.S. Pat. No. 6,579,968.

[0007] Despite the intense scientific and pharmaceutical interest in melanocortin receptor-specific peptides, evidenced by numerous articles in the scientific literature and numerous patent applications and issued patents, the only melanocortin receptor-specific peptide drugs approved in the United States are bremelanotide, sold under the trade name Viressi®, indicated for hypoactive sexual desire disorder in premenopausal women; afamelanotide, sold under the trade name SCENESSE®, indicated for the prevention of phototoxicity in adult patients with erythropoietic protoporphyria; and setomelanotide, sold under the trade name INCIVREE™, indicated for the treatment of obesity due to proopiomelanocortin (POMC), proprotein convertase subtilisin / kexin type 1 (PCSK1), or leptin receptor (LEPR) deficiency. There remains a significant need for melanocortin receptor-specific peptides for pharmaceutical applications, particularly for use in the treatment of inflammation-related diseases, indications, conditions, and syndromes. It is against this background that the present invention was made. Summary of the Invention [Means for solving the problem]

[0008] Brief Summary of the Invention In one aspect, the present invention provides a compound of formula I: [ka] (In the formula, Xaa 1 is -R5-R6; Xaa 2 and Xaa 3 are each independently 1 to 3 L- or D-isomer amino acids or combinations thereof linked by backbone amide bonds, wherein any backbone nitrogen atom is optionally alkylated with a C1 to C8 straight or branched alkyl, and when optionally alkylated, said alkyl optionally forms a ring with a side chain of the same amino acid when said side chain is aliphatic, said ring being optionally substituted; R1 is H or an L- or D-isomer amino acid side chain; R2 is -(CH2) u -or-(CH2) v -O-(CH2) w - and; R3 is H or a C1-C9 straight or branched aliphatic chain optionally containing one or more C=C double bonds; R4 is —H or C1-C8 linear or branched alkyl; R5 is optionally present, and when present, is 1 to 3 L- or D-isomer amino acids or combinations thereof linked by backbone amide bonds, any backbone nitrogen atom optionally being methylated; R6 is H or C1-C1, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl, or heteroaryl. 17 is an acyl group; x is 1 to 5; u is 2 to 8; v is 2 to 5; and (w is 2 to 5) or a pharmaceutically acceptable salt of any of the foregoing.

[0009] In the peptide of Formula I, Xaa 2 -Xaa 3may comprise the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted. 2 -Xaa 3 may comprise or consist of the amino acid sequence -His-D-Phe-Arg- or -His-Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted, and R is the side chain of the L- or D-isomer amino acid Trp.

[0010] In another embodiment, R1 can be the side chain of the L- or D-isomer amino acid Trp.

[0011] In another embodiment, Xaa 2 -Xaa 3 may comprise the amino acid sequence -His-D-Phe- or -His-Phe-, where the phenyl ring of D-Phe or Phe is optionally substituted, and R1 is the side chain of the L- or D-isomer amino acid Arg.

[0012] In another embodiment of the peptide of Formula I, R5 can be the L- or D-isomer of Nle or Arg. 2 -Xaa 3 may comprise the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted. Alternatively, in this aspect, Xaa 2 -Xaa 3 may comprise the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted, and R1 is the side chain of the L- or D-isomer amino acid Trp.

[0013] In another embodiment of the peptide of Formula I, R3 is absent and R4 is C1-C 17 In this embodiment, Xaa is an acyl group. 2 -Xaa 3 may comprise the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted.2 -Xaa 3 may comprise the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted, and R1 is the side chain of the L- or D-isomer amino acid Trp.

[0014] In another embodiment of the peptide of Formula I, R5 is an L- or D-isomer of Nle or Arg, and R6 is a C1-C 17 In this embodiment, Xaa is an acyl group. 2 and Xaa 3 comprises the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted. 2 and Xaa 3 may comprise the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, where the phenyl ring of D-Phe or Phe is optionally substituted, and R1 is the side chain of the L- or D-isomer amino acid Trp.

[0015] In other embodiments where the phenyl ring of D-Phe or Phe is optionally substituted, it is optionally substituted with 1 to 3 ring substituents, which may be the same or different and each independently include halo, (C1-C 10 ) Alkyl-halo, (C1-C 10 )Archi (C1~C 10 ) alkoxy, (C1-C 10 ) alkylthio, aryl, (C1-C 10 ) alkylaryl, aryloxy, nitro, nitrile, sulfonamido, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy-carbonyl or aryloxy-carbonyl.

[0016] In the peptides of Formula I, the L- or D-isomer amino acids can consist of alpha amino acids, beta amino acids, gamma amino acids, or delta amino acids, or combinations thereof.

[0017] Formula II: [ka] (In the formula, Xaa 1 is -R5-R6; R1 is substituted or unsubstituted indole, phenyl, or naphthyl; R2 is -(CH2) u -or-(CH2) v -O-(CH2) w - and; R3 is H or a C1-C9 straight or branched aliphatic chain optionally containing one or more C=C double bonds; R4 is -H or -CH3; R5 is optionally present, and when present, is 1 to 3 L- or D-isomer amino acids or combinations thereof, any backbone nitrogen atom optionally being methylated; R6 is H or C1-C1, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl, or heteroaryl. 17 is an acyl group; R7 is -H, -CH3 or -CH2-, and when it is -CH2-, together with R8 forms the general structure [ka] forming a ring of; R8 is -H when R8 forms a ring with R7, or R8 is -(CH2)3, -N(R 12a )(R 12b ), -NH-(CH2) z -N(R 12a )(R 12b ), -C(=O)-N(R 12a )(R 12b ), -O-(R 12a ), -S-(=O)2-CH3, -S-(=O)-CH3, substituted or unsubstituted phenyl, -O-CH2-phenyl (phenyl is substituted or unsubstituted), [ka] and; R9 is substituted or unsubstituted phenyl or naphthyl; R 10 teeth, -N(R 12a )(R 12b ), -NH-(CH2) z -N(R 12a )(R 12b ), -NH-C(=NH)-N(R 12a )(R 12b ), -NH-C(=O)-N(R 12a )(R 12b ), -O(R 12a ), -C1~C 17 linear, branched or cyclic alkyl chains, -S(=O)2-CH3, -S(=O)-CH3, -C(=O)-O(R 12a ), [ka] and; R 11 is —O—CH2-phenyl, where phenyl is substituted or unsubstituted; R 12a and R 12b is independently at each occurrence and in each instance H or a C1-C4 straight, branched or cyclic alkyl chain; t is independently in each instance 1 to 4; x is 1 to 5; u is 2 to 8; v and w are each independently 2 to 5; and z is 1 to 3. Further provided is a peptide of Formula I, including all enantiomers, stereoisomers or diastereomers thereof, or a pharmaceutically acceptable salt of any of the foregoing.

[0018] In the peptide of Formula II, R9 can be unsubstituted naphthyl.

[0019] In another aspect, in the peptide of Formula II, any substituted phenyl or naphthyl can be substituted, independently in each instance, with 1 to 3 ring substituents, which can be the same or different and each independently include halo, (C1-C 10 ) Alkyl-halo, (C1-C 10 ) alkyl, (C1-C 10 ) alkoxy, (C1-C 10 ) alkylthio, aryl, (C1-C 10 ) alkylaryl, aryloxy, nitro, nitrile, sulfonamido, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy-carbonyl or aryloxy-carbonyl.

[0020] In another embodiment of the peptide of Formula II, R5 comprises at least one L- or D-isomer amino acid and can be a single L- or D-isomer amino acid having an aliphatic side chain, including those in which the aliphatic side chain is -(CH2)3-CH3. Alternatively, in the peptide of Formula II, R5 can be a single L- or D-isomer amino acid having a side chain containing at least one nitrogen atom. R5 may be an amino acid, including, but not limited to, the L- or D-isomer of Arg, Lys, Orn, Dab, Dap, or Cit.

[0021] In another embodiment of the peptide of formula II, the peptide has the formula: [ka] It can be of the following type.

[0022] In any of the embodiments of the peptide of Formula II, Xaa 1 can be Ac-Arg, Ac-D-Arg, Ac-Nle, Ac-D-Nle, Ac-Trp, cyclopentylacetyl, hexanoyl, 3-phenylpropanoyl, cyclopentylacetyl, acetyl, 2-naphthylacetyl, or 3-(1-naphthyl)propanoyl; R can be indole or naphthalene; R and R together can comprise pyrrolidine, including those in which the pyrrolidine is substituted, including, but not limited to, those in which the substituent is -O-CH-phenyl, where the phenyl is substituted or unsubstituted; R can be substituted phenyl.

[0023] In an alternative embodiment, the peptide of formula I has formula III: [ka] (In the formula, Xaa 1 is -R5-R6; R1 is -N(R 12a )(R 12b ), -NH-(CH2) z -N(R 12a )(R 12b ), -NH-C(=NH)-N(R 12a )(R 12b ), -NH-C(=O)-N(R 12a )(R 12b ), -O(R 12a ), -C1~C 17 linear, branched or cyclic alkyl chains, -S(=O)2-CH3, -S(=O)-CH3, -C(=O)-O(R 12a ), [ka] and; R2 is -(CH2) u -or-(CH2) v -O-(CH2) w - and; R3 is H or a C1-C9 straight or branched aliphatic chain optionally containing one or more C=C double bonds; R4 is -H or -CH3; R5 is optionally present, and when present, is 1 to 3 L- or D-isomer amino acids or combinations thereof, any backbone nitrogen atom optionally being methylated; R6 is H or C1-C1, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl, or heteroaryl. 17 is an acyl group; R7 is -H, -CH3 or -CH2-, and when it is -CH2-, together with R8 forms the general structure [ka] forming a ring of; R8 is -H when R8 forms a ring with R7, or R8 is -H, -(CH2)3, -N(R 12a )(R 12b ), -NH-(CH2) z -N(R 12a )(R 12b ), -C(=O)-N(R 12a )(R 12b ), -O-(R 12a ), -S-(=O)2-R 12a , substituted or unsubstituted phenyl, -O-CH2-phenyl (phenyl is substituted or unsubstituted), [ka] and; R9 is substituted or unsubstituted phenyl or naphthyl; R 11 is —O—CH2-phenyl, where phenyl is substituted or unsubstituted; R 12a and R 12b is independently at each occurrence and in each instance H or a C1-C4 straight, branched or cyclic alkyl chain; t is independently in each instance 1 to 4; x is 1 to 5; u is 2 to 8; v and 2 are each independently 2 to 5; and z is 1 to 3. including all enantiomers, stereoisomers or diastereomers thereof or a pharmaceutically acceptable salt of any of the foregoing.

[0024] In one embodiment of the cyclic peptides of Formula III, R9 is unsubstituted naphthyl. In another embodiment of the peptides of Formula III, either substituted phenyl or naphthyl can be substituted, independently at each instance, with 1 to 3 ring substituents, which can be the same or different and each independently include halo, (C1-C 10 ) Alkyl-halo, (C1-C 10 ) alkyl, (C1-C 10 ) alkoxy, (C1-C 10 ) alkylthio, aryl, (C1-C 10 ) Alkylaryl, aryloxy, nitro, nitrile, sulfonamide, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy It is aryloxy-carbonyl or aryloxy-carbonyl.

[0025] In another embodiment of the peptide of Formula III, R5 can include at least one L- or D-isomer amino acid, which can be a single L- or D-isomer amino acid having an aliphatic side chain.

[0026] In another embodiment of the peptide of Formula III, R6 can be acetyl, cyclopentylacetyl, hexanoyl, 3-phenylpropanoyl, cyclopentylacetyl, 2-naphthylacetyl, or 3-(1-naphthyl)propanoyl.

[0027] In another embodiment of the peptide of formula III, the peptide has the formula: [ka] It is a cyclic peptide of the formula:

[0028] In another embodiment, the present invention provides a compound of formula IV or V: [ka] (In the formula, Z is H or an N-terminal group; Xaa 1 is optionally present, and when present, is 1 to 3 amino acids, any backbone nitrogen atom of which is optionally methylated; Xaa 2 is the group -NH-(CH2) u -NH- or -NH-(CH2) v -O-(CH2) w an L- or D-isomer of an amino acid having a side chain containing a carboxyl group that forms an amide bond with the first -NH- of the -NH-; Xaa 3 is an L- or D-isomer amino acid of Pro optionally substituted with hydroxyl, halogen, sulfonamide, alkyl, -O-alkyl, aryl, alkyl-aryl, alkyl-O-aryl, alkyl-O-alkyl-aryl, -O-alkyl-aryl or -O-aryl, or Xaa 3is at least one primary amine, secondary amine, alkyl, cycloalkyl, cycloheteroalkyl, aryl, L- or D-isomer amino acids with heteroaryl, ether, sulfide, or carboxyl-containing side chains; Xaa 4 is an L- or D-isomer amino acid having a substituted or unsubstituted aryl-containing side chain; Xaa 5 is an L- or D-isomer amino acid having a side chain containing at least one primary amine, secondary amine, guanidine, urea, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, or ether, and Xaa 6 is absent, the C-terminal carboxyl group is the group -NH-(CH) u -NH- or -NH-(CH2) v -O-(CH2) w -NH- forms an amide bond with the second -NH-; Xaa 6 is optionally present, and if present, is an L- or D-isomer amino acid having a side chain containing at least one aryl or heteroaryl optionally substituted with one or more ring substituents, where one or more, if present, are the same or different and independently are hydroxyl, halogen, sulfonamide, alkyl, -O-alkyl, aryl, or -O-aryl, and the C-terminal carboxyl group is selected from the group -NH-(CH) u -NH- or -NH-(CH2) v -O-(CH2) w -NH- forms an amide bond with the second -NH-; u is 2 to 8; v and w each independently represent 2 to 5; Group -NH-(CH2) u -NH- and -NH-(CH2) v -O-(CH2) w -NH- includes a first -NH- and a second -NH-. or a pharmaceutically acceptable salt thereof.

[0029] In the cyclic peptides of formula IV or V, Z is a C1-C alkyl group, including linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, or aralkyl. 17 The N-terminal group may be selected from the group consisting of acyl groups.

[0030] In the cyclic peptide of formula IV or V, Xaa 1 can be a single amino acid residue selected from the group consisting of Gly or the L- or D-isomers of Ala, Nle, Leu, Ile, or Val. 1 can be a single amino acid with a side chain containing at least one primary amine, guanidine, or urea group, including, but not limited to, the L- or D-isomers of Arg, Lys, Orn, Dab, Dap, or Cit.

[0031] In the cyclic peptide of formula IV or V, Xaa 4 can be D-Phe or Phe optionally substituted with 1 to 3 ring substituents. The ring substituents can be the same or different and each independently include halo, (C1-C 10 ) Alkyl-halo, (C1-C 10 ) alkyl, (C1-C 10 ) alkoxy, (C1-C 10 ) alkylthio, aryl, (C1-C 10 ) alkylaryl, aryloxy, nitro, nitrile, sulfonamido, amino, monosubstituted amino, disubstituted amino, hydroxy, carboxy, or alkoxy-carbonyl. 4 can be D-Nal1 or D-Nal2.

[0032] In the cyclic peptide of formula IV or V, Xaa 5 can be the L- or D-isomer of Arg, Lys, Orn, Dab, or Dap, and Xaa 6 can be the L- or D-isomer of Trp, Na11, or Na12.

[0033] Cyclic peptides of formula IV or V therefore include, but are not limited to, the following substituents: Z is a C1-C7 linear alkyl acyl group; Xaa 1 is the L- or D-isomer of Nle or Arg; Xaa 2 is the L- or D-isomer of Asp or Glu, and the side chain carboxyl group is the group -NH-(CH) u -NH- or -NH-(CH2) v -O-(CH2) w -NH- forms an amide bond with the first -NH-; Xaa 3 is the L- or D-isomer of His, Hyp(Bzl), Met(O2) or Asn; Xaa 4 is the L- or D-isomer of substituted or unsubstituted Phe, Na1 or Na12; Xaa 5 is the L- or D-isomer of Arg; and Xaa 6 is the L- or D-isomer of Trp, Na1 or Na12, the C-terminal carboxyl group of which is a group -NH-(CH2) u -NH- or -NH-(CH2) v -O-(CH2) w The -NH- forms an amide bond with the second -NH-.

[0034] Further provided are embodiments of cyclic peptides of Formula IV, wherein Xaa 2 The side chain carboxyl group of is the group -NH-(CH2) u -NH of Xaa forms an amide bond with the first -NH- of Xaa 6 The C-terminal carboxyl group of is the group -NH-(CH2) u The -NH- forms an amide bond with the second -NH-.

[0035] In another embodiment of the cyclic peptide of Formula IV or V, at least one backbone nitrogen atom thereof may comprise a methyl group. In an alternative embodiment, none of the backbone nitrogen atoms thereof comprises a methyl group.

[0036] In another embodiment, the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0037] In the aforementioned cyclic peptides, the pharmaceutically acceptable salt may be an acetate salt or alternatively a trifluoroacetate salt.

[0038] In another embodiment, [ka] or a pharmaceutically acceptable salt thereof.

[0039] Further provided is a pharmaceutical composition comprising a cyclic peptide as set forth above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0040] The present invention further includes a method for treating a melanocortin receptor-mediated disease, indication, condition, or syndrome in a human or non-human mammal, the method comprising administering a pharmaceutical composition comprising one or more cyclic peptides as set forth above. The present invention further includes a method for treating a condition responsive to altered melanocortin receptor function in a human or non-human mammal, the method comprising administering a pharmaceutical composition comprising one or more cyclic peptides as set forth above. In the method of treatment, the melanocortin receptor-mediated disease, indication, condition, or syndrome in a human may be ocular inflammation, and the pharmaceutical composition may be an aqueous pharmaceutical composition for administration to the ocular surface.

[0041] Ocular inflammatory responses to altered melanocortin receptor function can be caused by ocular conditions, including dry eye disease, corneal ulcers, corneal erosions, corneal abrasions, corneal degeneration, corneal perforation, corneal scarring, epithelial defects, keratoconjunctivitis, idiopathic uveitis, corneal transplants, age-related macular degeneration, diabetic eye pathology, blepharitis, glaucoma, ocular hypertension, postoperative ocular pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascular membranes, vascular occlusive disease, and ocular conditions. The inflammation may be caused by dry eye disease or keratoconjunctivitis sicca, inflammatory eye disease, ocular surface disease, corneal disease, retinal disease, ocular manifestations of systemic disease, inherited eye conditions, ocular tumors, elevated intraocular pressure, herpetic infections, pterygium, persistent wounds to the surface of the eye, ocular pain and inflammation after laser photorefractive keratectomy, thermal or chemical burns to the cornea, scleral wounds, keratoconus, or conjunctival wounds. In one aspect, the inflammation is caused by dry eye disease or keratoconjunctivitis sicca.

[0042] In one embodiment, a pharmaceutical composition for ocular administration is provided, wherein the pharmaceutically acceptable carrier is an aqueous solution containing about 2.79 mg / mL trisodium citrate dihydrate, about 9 mg / mL sodium chloride, and about 1 mg / mL polysorbate 80. In this pharmaceutical composition, the cyclic peptide may be in the form of a trifluoroacetate salt at a concentration of about 1.0 μg / mL. The pharmaceutical composition may further contain about 0.10 mg / mL anhydrous citric acid. The aqueous solution of the pharmaceutical composition has a pH of about 6.5, and the composition further contains sodium hydroxide or hydrochloric acid as needed to adjust the pH. In a further aspect, the present invention provides a method for treating dry eye disease or keratoconjunctivitis sicca, comprising administering about 50 μL or less of the pharmaceutical composition no more than about three times per day. Preferably, no more than about 150 ng of the cyclic peptide or a pharmaceutically acceptable salt thereof is administered per eye per day.

[0043] In a related aspect, the present invention provides a method for treating dry eye disease or keratoconjunctivitis sicca, comprising administering to the ocular surface, in divided doses administered no more than three times per day, no more than about 150 ng per eye per day, with an EC of less than 1 nM for each of the MC1r and MC5r. 50 The present invention provides a method for treating rheumatoid arthritis, comprising administering a melanocortin receptor-specific peptide having an EC value of 1.0, as determined by cAMP production using HTRF detection methods such as those performed by CEREP. The administered dose can be less than 150 ng / eye / day of the peptide of the present invention, and can be about 100 ng / eye / day or about 50 ng / eye / day of the peptide of the present invention. The peptide has an EC value of 1.0, as determined by cAMP production using HTRF detection methods such as those performed by CEREP. 50 In one aspect, the melanocortin receptor-specific peptide is an agonist or partial agonist at each of the MC1r and MC5r, having a cytotoxicity value of: [ka] It is a cyclic peptide of the formula:

[0044] In one aspect, a cyclic peptide template is provided that can be utilized in the generation of receptor-specific peptides for biological receptors.

[0045] In another aspect, pharmaceutical compositions based on melanocortin receptor-specific peptides are provided for use in the treatment of melanocortin receptor-mediated diseases, indications, conditions and syndromes.

[0046] In another aspect, peptide-based melanocortin receptor-specific pharmaceuticals are provided, which are selective and are agonists at MC1r and antagonists at MC4r.

[0047] In another aspect, a peptide-based melanocortin receptor-specific pharmaceutical is provided, which is selective and is an agonist at MC1r and a partial agonist at MC4r.

[0048] In another embodiment, a sub-nanomolar EC50 Receptor-specific peptides are provided that are functionally active at one or more melanocortin receptors at a specific level.

[0049] In another embodiment, an EC 50 Melanocortin receptor-specific peptides that are agonists or partial agonists at the MC1r and MC5r receptors are provided.

[0050] Other aspects and novel features and further scope of applicability of the present invention will be set forth in part in the detailed description which follows, 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. Aspects of the present invention may be realized and attained by means of the instrumentalities and combinations particularly pointed out in the appended claims.

[0051] BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and form a part of this specification, illustrate one or more embodiments of the invention and, together with the description, serve to explain the principles of the invention. The drawings are intended merely to illustrate one or more preferred embodiments of the invention and are not to be construed as limiting the invention. [Brief explanation of the drawings]

[0052] [Figure 1A] Box / whisker plot showing the median (line), 25th / 75th percentile (box), mean (+) and (whisker) corneal fluorescein scores on day 12 after exposure to a controlled dry environment with transdermal scopolamine to establish disease before any treatment. Individual data points are shown as filled circles. The number of eyes (n) was 24 per group, except for the untreated group (n=14). [Figure 1B] 1B is a box / whisker plot showing the median (line), 25th / 75th percentile (box), mean (+) and (whisker) corneal fluorescein scores on day 24 after treatment initiated on day 13 for the group in FIG. 1A. Individual data points are shown as filled circles. *P<0.05, **P<0.01 and ***P<0.001 relative to untreated eyes. [Figure 2] Box and whisker plots of the data as in Figures 1A and 1B show treatment comparisons between day 12 (before treatment) and day 24 (12 days after topical treatment with test compound) for untreated eyes, PBS (vehicle control phosphate buffered saline), and each test compound. Administration of Peptide No. 60 at doses of 10-2 to 10-4 mg / mL resulted in statistically significant improvements in corneal fluorescein scores from day 12 to day 24. DETAILED DESCRIPTION OF THE INVENTION

[0053] Detailed Description of the Invention 1.0 Definition Before proceeding with the description of the present invention, certain terms are defined as set forth herein.

[0054] In the sequences provided for the peptides disclosed herein, the amino acid residues are as follows: th Those as given in Chapter 2400 of Ed. and have their conventional meanings. Thus, "Ala" is alanine, "Asn" is asparagine, "Asp" is aspartic acid, "Arg" is arginine, "Cys" is cysteine, "Gly" is glycine, "Gln" is glutamine, "Glu" is glutamic acid, "His" is histidine, "Ile" is isoleucine, "Leu" is leucine, "Lys" is lysine, "Met" is methionine, "Phe" is phenylalanine, "Pro" is proline, "Ser" is serine, "Thr" is threonine, "Trp" is tryptophan, "Tyr" is tyrosine, "Val" is valine, etc. It is understood that the D-isomer is indicated by the three-letter code or "D-" before the amino acid name, e.g., D-Phe becomes D-phenylalanine. Amino acid residues not encompassed by the foregoing include, but are not limited to, those having the following side chains, it being understood that such amino acid residues may be either the L- or D-isomer.

[0055] [Table 1]

[0056] [Table 2]

[0057] [Table 3]

[0058] [Table 4]

[0059] [Table 5]

[0060] Amino acid residues further include, but are not limited to, the following, and it is understood that such amino acid residues may be either the L- or D-isomer.

[0061] [Table 6]

[0062] The term "alpha amino acid" refers to the general structure [ka] (shown in its non-ionized form), where R is any side chain group or hydrogen, including, but not limited to, the amino acid residues or side chain groups set forth in the preceding tables and paragraphs.

[0063] The terms "L- or D-isomer amino acid" or "L- or D-isomer amino acid(s)" include all isomeric forms of any amino acid residue as defined herein, specifically including, but not limited to, any alpha amino acid, beta amino acid, gamma amino acid, or delta amino acid, those encoded directly by DNA, post-translationally modified amino acids, amino acids expressed by biological means other than directly by DNA, proteinogenic or non-proteinogenic amino acids, or any synthetic or artificial amino acid.

[0064] Amino acids, including L- or D-isomer amino acids, are linked together by "amide bonds" or amide linkages to form covalent peptide bonds that connect the backbone carboxylic acid group of one amino acid with the backbone amino group of another amino acid, thereby forming a peptide bond (-C(=O)-NH-) or backbone amide bond.

[0065] The term "acyl" includes the group R(C=O)-, where R is an organic group, such as alkyl, aryl, heteroaryl, carbocyclyl, or heterocyclyl. Reference herein to a substituted acyl group means that the organic group (R) is substituted. Non-limiting examples of acyl groups include CH3-C(=O)-, sometimes referred to herein as acetyl or "Ac"; CH3-(CH2)4-C(=O)-, sometimes referred to herein as hexanoyl or "Hex"; CH3-(CH2)5-C(=O)-, sometimes referred to herein as heptanoyl or "Hept"; and various cyclyl groups, such as phenylpropanoyl and cyclopentylacetyl.

[0066] A peptide or aliphatic moiety is acylated when an aliphatic or substituted aliphatic group or an aromatic-substituted aromatic group is attached through a carbonyl (-(C=O)-) group to form an acyl group. Peptides are most often acylated at the N-terminus.

[0067] The term "alkane" includes linear or branched saturated hydrocarbons. Examples of linear alkane groups include methane, ethane, propane, etc. Examples of branched or substituted alkane groups include methylbutane or dimethylbutane, methylpentane, dimethylpentane, or trimethylpentane, etc. In general, any alkyl group can be a substituent of an alkane.

[0068] The term "alkene" includes unsaturated hydrocarbons containing one or more double carbon-carbon bonds. Examples of such alkene groups include ethylene, propene, and the like.

[0069] The term "alkenyl" includes linear monovalent hydrocarbon radicals of two to six carbon atoms or branched monovalent hydrocarbon radicals of three to six carbon atoms containing at least one double bond; examples thereof include ethenyl, 2-propenyl, and the like.

[0070] The "alkyl" groups specified herein include alkyl groups of designated length that are either straight- or branched-chain saturated aliphatic hydrocarbon groups. 1~10 Alkyl means an alkyl having 1 to 10 carbon atoms. Non-limiting examples of such alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tertiary butyl, pentyl, isopentyl, hexyl, isohexyl, and the like.

[0071] The term "alkyne" includes a linear monovalent hydrocarbon radical of 2 to 6 carbon atoms or a branched monovalent hydrocarbon radical of 3 to 6 carbon atoms containing at least one triple bond; examples include ethyne, propyne, butyne, and the like.

[0072] The term "aryl" includes monocyclic or bicyclic aromatic hydrocarbon groups of 6 to 12 ring atoms, optionally substituted with one or more substituents independently selected from alkyl, haloalkyl, cycloalkyl, alkoxy, alkthio, halo, nitro, acyl, cyano, amino, monosubstituted amino, disubstituted amino, hydroxy, carboxy, or alkoxy-carbonyl. Examples of aryl groups include phenyl, biphenyl, naphthalene, naphthyl, 1-naphthyl, and 2-naphthyl, derivatives thereof, and the like. Similarly, the term "naphthyl" includes 1-naphthyl and 2-naphthyl, and "naphthalene" includes 1-naphthalene and 2-naphthalene.

[0073] The term "aralkyl" refers to the group -R a R b (In the formula, R a contains an alkylene (divalent alkyl) group, and R b is an aryl group as defined above. Examples of aralkyl groups include benzyl, phenylethyl, 3-(3-chlorophenyl)-2-methylpentyl, and the like.

[0074] The term "aliphatic" includes compounds that have a hydrocarbon chain, such as alkyl, aryl, heteroaryl, alkane, alkene, alkyne, and derivatives thereof.

[0075] As used herein, the term "amide" refers to a compound having a trivalent nitrogen linked to a carbonyl group, i.e., -C(=O)-NH2 (i.e., primary amide), -C(=O)-NHR c and -C(=O)-NR c R d (In the formula, R c and R d each independently represents hydrogen or an organic group). When a substituted amide group is referred to herein, the organic group (R c and R d ) is substituted. Examples of amide include methylamide, ethylamide, and propylamide.

[0076] "Amine" includes amino groups (-NH2), -NHR a and -NR a R b (In the formula, R a and R b each independently represents hydrogen or an organic group). When referring to a substituted amine group herein, the organic group (R a and R b ) is substituted.

[0077] "Nitriles" include compounds that are carboxylic acid derivatives and contain a (-CN) group attached to an organic group.

[0078] The term "halogen" is intended to include the halogen atoms fluorine, chlorine, bromine, and iodine, as well as groups containing one or more halogen atoms, such as, for example, --CF.sub.3.

[0079] The term "composition," as in pharmaceutical composition, is intended to encompass a product containing an active ingredient and inactive ingredients constituting a carrier, as well as any product resulting directly or indirectly from the combination, complexation, or aggregation of any two or more of the ingredients, or from the dissociation of one or more of the ingredients, or from any other type of reaction or interaction of one or more of the ingredients. Thus, a pharmaceutical composition is prepared by mixing an active ingredient with one or more pharmaceutically acceptable carriers. This includes any composition prepared by

[0080] A melanocortin receptor "agonist" refers to an endogenous substance, drug substance, or compound, including certain of the peptide compounds disclosed herein, that can interact with a melanocortin receptor and initiate a pharmacological response, including but not limited to, receptor activation, including initiating signal transduction characteristic of the melanocortin receptor, such as adenyl cyclase activation. A melanocortin receptor agonist can be an agonist at one or more of the MC1r, MC2r, MC3r, MC4r, and MC5r.

[0081] A melanocortin receptor "antagonist" refers to an endogenous substance, drug substance, or compound, including certain of the peptide compounds disclosed herein, that blocks or attenuates the action of an agonist at a melanocortin receptor. A melanocortin receptor antagonist can be an antagonist at one or more of the MC1r, MC2r, MC3r, MC4r, and MC5r. Certain compounds, including certain of the peptide compounds disclosed herein, can be agonists at one or more melanocortin receptors and antagonists at one or more other melanocortin receptors.

[0082] "α-MSH" means the peptide Ac-Ser-Tyr-Ser-Met-Glu-His-Pro-Arg-Glu-Lys-Pro-Val-NH and analogs and homologs thereof, including, but not limited to, NDP-α-MSH.

[0083] "NDP-α-MSH" means the peptide Ac-Tyr-Ser-Tyr-Ser-Nle-Glu-His-DlE-Pro-Arg-Glu-Lys-Pro-Val-NH and analogs and homologs thereof.

[0084] "EC 50 " means the molar concentration of an agonist, including partial agonists, that produces 50% of the maximum possible response for that agonist. As an example, the EC50 of a test compound that produces 50% of the maximum possible response for that compound at a concentration of 72 nM as determined in a cAMP assay in an MC4r cell expression system. 50 is 72 nM. Unless otherwise specified, EC 50 The molar concentration relevant for determination is nanomoles per liter (nM).

[0085] "Ki (nM)" refers to the equilibrium inhibitor dissociation constant, which represents the molar concentration of a competitive compound that binds to half of the receptor's binding sites at equilibrium in the absence of a competitor. Generally, the value of Ki is inversely related to the affinity of a compound for receptor, and the lower Ki indicates the higher affinity. Ki can be determined using the Cheng and Prusoff formula (Cheng Y., Prusoff WH, Biochem. Pharmacol. 22: 3099-3108, 1973).

number

[0086] Unless otherwise specified, molar concentrations associated with Ki determinations are in nM. Ki can be expressed with respect to a specific receptor (e.g., MC1r, MC3r, MC4r, or MC5r).

[0087] "Inhibition" refers to the percent attenuation or reduction in receptor binding in a competitive inhibition assay compared to a known standard. Thus, "inhibition at 1 μM (NDP-α-MSH)" refers to the percent reduction in binding of NDP-α-MSH upon addition of a determined amount of the compound to be tested, such as 1 μM of test compound, under the assay conditions described herein below. By way of example, a test compound that does not inhibit binding of NDP-α-MSH will have an inhibition of 0%. and the inhibition of a test compound that completely inhibits the binding of NDP-α-MSH is 100%. Generally, as described herein below, 125 For competitive inhibition tests using, for example, NDP-α-MSH or lanthanide chelate fluorescence assays using, for example, Eu-NDP-α-MSH, a detectably labeled assay is used. However, other methods of testing competitive inhibition are known, including the use of different label or tag systems, and generally, any method known in the art for testing competitive inhibition can be used in the present invention. Thus, "inhibition" can be seen to be one measure for determining whether a test compound attenuates the binding of α-MSH to the melanocortin receptor.

[0088] "Binding affinity" means the ability of a compound or drug to bind to its biological target, and is expressed herein as Ki (nM).

[0089] "E max " means the maximum functional activity achievable by a compound in a specified melanocortin receptor-expressing cell line, such as maximal stimulation of adenylyl cyclase. The maximal stimulation achieved by NDP-α-MSH is 100% of the "E max " and the E of compounds that can stimulate half of the maximum activity of NDP-α-MSH. max is considered to be 50%. An E of 70% or greater under the assay conditions described herein max Compounds of the invention having the formula: max Compounds with an E of 10% to 70% can be classified as partial agonists, max Compounds with a β-amylindrical activity below 10% can be classified as inactive.

[0090] Generally, "functional activity" is a measure of receptor signaling or a change in receptor-associated signaling, such as by a melanocortin receptor, upon activation of the receptor by a compound. Melanocortin receptors initiate signaling through activation of heterotrimeric G proteins. In one aspect, melanocortin receptors activate Gα , which catalyzes the production of cAMP by adenylyl cyclase. S Signals are transmitted through adenylyl cyclase. Thus, determining stimulation of adenylyl cyclase, such as determining maximal stimulation of adenylyl cyclase, is one measure of functional activity and is the primary measure exemplified herein. However, it is understood that alternative measures of functional activity can be used in the practice of the present invention and are specifically contemplated and within the scope of the present invention. Thus, in one example, intracellular free calcium can be measured using a specific fluorescent molecule binding to calcium, such as Fura2, as disclosed and reported using the methods disclosed in Mountjoy KG et al., Melanocortin receptor-medicated mobilization of intracellular free calcium in HEK293 cells. Physiol Genomics 5: 11-19, 2001 or Newman et al., Activation of the melanocortin-4 receptor mobilizes intracellular free calcium in immortalized hypothalamic neurons. J Surg Res: 132: 201-207, 2006. Fluo-4 is an alternative calcium-binding dye that is commonly used (Nohr et al., The orphan G protein-coupled receptor GPR139 is activated by the peptides: Adrenocorticotropic hormone(ACTH), α-, and β-melanocyte stimulating hormone (α-MSH, and β-MSH), and the conserved core motif HFRW. Neurochem Int 102: 105-113, 2017). Ca2 + For release events and further in the same pathway Activation can also be measured upstream by measuring the production of inositol triphosphate or diacylglycerol from phosphatidylinositol 4,5-bisphosphate, such as by commercially available HTRF assays. (Liu et al., Comparison on functional assays for Gq-coupled GPCRs by measuring inositol monophospate-1 and intracellular calcium in a 1536-well plate format. Curr Chem Genomics 1: 70-77, 2008). Another measure of functional activity is the result of regulatory pathway activation, using methods such as those disclosed in Nickolls SA et al., Functional selectivity of melanocortin 4 receptor peptide and nonpeptide agonists: evidence for ligand-specific conformational states. J Pharm Exper Therapeutics 313: 1281-1288, 2005. Another measure of functional activity is the exchange and exchange rate of nucleotides associated with activation of G protein receptors, e.g., G protein α subunits. The conversion of GDP (guanosine diphosphate) to GTP (guanosine triphosphate) in the guanosine 5'-(γ-[ 35 It can be measured by several means, including radioassays using [S]thio)-triphosphate. A relatively new assay platform has been invented to measure the activity / coupling of 14 different Gα species belonging to the Gi, Gq, Gs, and Gi2 / i3 subfamilies as they relate to receptors, using biosensors based on BRET (bioluminescence resonance energy transfer) to measure disengagement of Gα and Gγ subunits upon ligand binding (Zhao et al., Biased signaling of protease-activated receptors. Front Endocrinol 5: 67, 2014; and van der Westhuizen et al., Quantification of ligand bias for clinically relevant β2-adrenergic receptor ligands: Implications for drug taxonomy. Molecular Pharm 85: 492-509, 2014). Various gene-based assays have been developed, such as Chen W. et al., A colorimetric assay from measuring activation of Gs- and Gq-coupled signaling pathways. Anal Biochem 226: 349-354, 1995; Kent TC et al., Development of a generic G-coupled proteins, such as those disclosed in "dual-reporter gene assay for screening G-protein-coupled receptors." Biomol Screening, 5: 437-446, 2005; or "Kotarsky K. et al., Improved receptor gene assays used to identify ligands acting on orphan seven-transmembrane receptors." Pharmacology & Toxicology 93: 249-258, 2003." The colorimetric assay of Chen et al. was developed to measure protein activation. Hruby VJ et al., Cyclic lactam α-melanocortin analogues of Ac-Nle 4 -cyclo[Asp 5 , D-Phe 7 , Lys 10] α-melanocyte-stimulating hormone-(4-10)-NH2with bulky aromatic amino As disclosed in J Med Chem 38: 3454-3461, 1995, melanocortin acids at position 7 show high antagonist potency and selectivity at specific melanocortin receptors. The present invention is suitable for use in measuring G-coupled receptor activation. Generally, functional activity can be measured by any method, including methods for determining G-coupled receptor activation and / or signal transduction, including methods that may be developed or reported herein below. Each of the aforementioned articles and methods disclosed herein are incorporated by reference as if fully set forth.

[0091] The terms "treat," "treating," and "treatment," as used herein, contemplate an action that occurs while a patient is suffering from the specified disease or disorder, which reduces the severity of the disease or disorder.

[0092] As used herein, the term "pharmaceutically effective amount" (including "therapeutically effective amount") means an amount of a peptide according to the present invention that is sufficient to induce a desired therapeutic or biological effect.

[0093] As used herein, the term "therapeutically effective amount" means an amount of a compound, including a peptide of the present invention, that elicits a biological or medical response in a mammal being treated by a physician or other clinician.

[0094] As used herein, the term "prophylactic effect" or "prophylactic" refers to an amount of a compound, including a peptide of the present invention, that prevents or inhibits or alleviates suffering in a mammal with a medical condition that a physician or other clinician seeks to prevent, inhibit, or alleviate before the patient begins to suffer from the specified disease or disorder.

[0095] 2.0 Clinical Indications and Usefulness The compositions and methods disclosed herein can be used for both medical and animal husbandry or veterinary applications. The term "patient" refers to a human, and as used herein and as used so throughout and in the claims. While the primary application of the peptides disclosed herein or of the formulas disclosed herein involves human patients, the peptides disclosed herein or of the formulas disclosed herein may be applied to laboratory, farm, zoo, wild, pet, sport or other animals. Clinical indications and specific utilities include:

[0096] 2.1 Inflammatory and fibrotic diseases and conditions The peptides and compositions of the present invention, including but not limited to peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be used in treating inflammatory diseases and conditions in patients. There are many inflammatory diseases and conditions that can be so treated. In one aspect, the inflammatory condition arises from diseases including arthritis, including but not limited to osteoarthritis, rheumatoid arthritis, septic arthritis, gout and pseudogout, juvenile idiopathic arthritis, Still's disease and ankylosing spondylitis, as well as arthritis secondary to other diseases, such as arthritis secondary to lupus erythematosus, Henoch-Schonlein purpura, psoriatic arthritis, reactive arthritis, hemochromatosis, hepatitis, Wegener's granulomatosis, vasculitis syndromes, Lyme disease, familial Mediterranean fever, hyperimmunoglobulinemia D with recurrent fever, TNF receptor-associated periodic syndrome, and forms of inflammatory bowel disease, including Crohn's disease and ulcerative colitis. In another embodiment, the inflammatory condition results from inflammatory bowel disease, including, for example, Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, fecal diversion colitis, Behcet's syndrome, infectious colitis, and forms of intractable colitis. In another embodiment, the inflammatory condition results from systemic syndromes, such as systemic lupus erythematosus, Sjogren's syndrome, scleroderma, rheumatoid arthritis, and polymyositis, or syndromes that affect only local body systems, such as, but not limited to, autoimmune diseases, including the endocrine system (type 1 diabetes, Hashimoto's thyroiditis, Addison's disease, etc.), dermatological system (pemphigus vulgaris), hematologic system (autoimmune hemolytic anemia), or nervous system (multiple sclerosis).Thus, autoimmune diseases include diseases and conditions such as acute disseminated encephalomyelitis, Addison's disease, ankylosing spondylitis, antiphospholipid syndrome, aplastic anemia, autoimmune hepatitis, autoimmune oophoritis, celiac disease, Crohn's disease, gestational pemphigoid, Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, idiopathic thrombocytopenic purpura, Kawasaki disease, lupus erythematosus, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, opsoclonus-myoclonus syndrome, optic neuritis, Ordo's thyroiditis, pemphigus, pernicious anemia, primary biliary cirrhosis, Reiter's syndrome, Sjogren's syndrome, Takayasu's arteritis, temporal arteritis, autoimmune hemolytic anemia, and Wegener's granulomatosis.

[0097] In another aspect, the inflammatory condition results from or is associated with chronic obstructive pulmonary disease (COPD), also known as chronic obstructive airway disease, including, but not limited to, diseases characterized by pathological airflow restriction in the airways that is not fully reversible, such as chronic bronchitis, emphysema, pneumoconiosis, lung neoplasms, and other lung disorders. Other inflammatory conditions include upper or lower respiratory tract diseases and disorders, such as allergic asthma, non-allergic asthma, nasal hyperreactivity, vasomotor rhinitis, allergic conjunctivitis, non-allergic conjunctivitis, and exotoxin- or substance-related airway diseases, such as various forms of pneumoconiosis (coal worker's pneumoconiosis, asbestosis, silicosis, bauxite fibrosis, beryllium disease, or siderosis), assinosis, or hypersensitivity pneumonitis (farmer's lung or bird breeder's disease). Other lung diseases involving inflammatory conditions include acute respiratory distress syndrome. The peptides and compositions of the invention are of particular utility for the treatment of conditions in which glucocorticoids are either ineffective or inadequate to produce the desired pharmacological response, such as COPD, asthma in smokers, and other conditions characterized in whole or in part by eosinophil accumulation in the lung, neutrophil infiltration and activation, alveolar macrophage recruitment and activation, epithelial cell expression of IL-8, or elevated expression of TNF-α. For airway or pulmonary disorders, in one embodiment, the peptides of the invention are delivered systemically; in another embodiment, the peptides of the invention are delivered locally, such as by inhalation.

[0098] In another aspect, the inflammatory condition results from or is associated with some form of transplant-related condition or syndrome, such as graft-versus-host disease, hyperacute rejection, acute rejection, or chronic rejection. Graft-versus-host disease is a common complication of allogeneic bone marrow transplantation, but it can also occur with other transplants, and particularly with T cells present in the graft, either as contaminants or intentionally introduced. Hyperacute, acute, or chronic rejection can occur with transplants of bodily organs such as kidney, liver, pancreas, spleen, uterus, heart, or lung, as well as bone, cornea, face, hand, penis, or skin. In one embodiment, a pharmaceutical composition comprising one or more peptides of the present invention is administered prophylactically to limit or prevent a transplant-related condition or syndrome, such as immediately before, during, or after transplantation of a bodily fluid, organ, or part. In another embodiment, the bodily fluid, organ, or part to be transplanted is perfused with a solution of a pharmaceutical composition comprising one or more peptides of the present invention. In yet another embodiment, one or more of the peptides of the invention are administered in conjunction with, in combination with, or in conjunction with one or more other agents against transplant rejection, such as calcineurin inhibitors including cyclosporine or tacrolimus, mTOR inhibitors including sirolimus or everolimus, antiproliferative agents including azathioprine or mycophenolic acid, corticosteroids including prednisolone or hydrocortisone, monoclonal anti-IL-2Rα receptor antibodies, antibodies such as basiliximab or daclizumab, or polyclonal anti-T cell antibodies such as antithymocyte globulin or antilymphocyte globulin.

[0099] In yet another aspect, the peptides and compositions of the present invention, including but not limited to peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be utilized in the treatment of fibrotic and sclerosing diseases, indications, conditions, and syndromes in patients. There are many fibrotic and sclerosing diseases, indications, conditions, and syndromes that can be so treated. Fibrotic and sclerosing diseases, indications, conditions, and syndromes often involve inflammatory factors, and therefore many can also be classified as inflammatory diseases or conditions, as listed above. In addition to involving an inflammatory component, fibrotic and sclerosing diseases and conditions can also be idiopathic, toxic, genetic, and / or pharmacologically induced disorders. In general, fibrotic disorders are characterized by the overproduction of extracellular matrix, primarily type I collagen, which can lead to loss of organ function. Without wishing to be bound by theory, it is believed that agonism of MC1r may result in the suppression of transforming growth factor-β1-induced collagen synthesis by human dermal fibroblasts, thereby providing therapeutic and / or prophylactic benefit for fibrotic and sclerosing diseases, indications, conditions, and syndromes. Exemplary fibrotic and sclerosing diseases and conditions that may be so treated include, but are not limited to, localized scleroderma, systemic sclerosis, cutaneous sclerodermatous graft-versus-host disease, idiopathic pulmonary fibrosis, bleomycin-induced pulmonary fibrosis, cyclosporine-induced nephropathy, cirrhosis of the liver, hypertrophic scars, keloids, and the like.

[0100] In yet another aspect, peptides and compositions of the present invention, including but not limited to peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, and particularly MC1r and MC5r agonists, can be used in treating fibrotic diseases, conditions, and syndromes in patients. Such fibrotic processes can be secondary to chronic inflammation, and the development of fibrosis is a common consequence of chronic inflammation. There are a wide variety of diseases in which fibrosis contributes to mortality and morbidity, including pulmonary fibrosis, liver fibrosis and cirrhosis, chronic kidney disease, myocardial infarction, and systemic autoimmune diseases such as systemic sclerosis. Fibrosis can also occur in eye diseases, particularly those characterized by chronic inflammation. The peptides and compositions of the present invention may inhibit the formation of fibrosis, possess regenerative properties, and are believed to reduce or ameliorate the effects of fibrosis.

[0101] In yet another embodiment, the peptides include peptides that are MC1r, MC3r, MC4r and / or MC5r agonists, partial agonists, antagonists, or any combination thereof. The peptides and compositions of the present invention, including but not limited to, can be utilized in the treatment of diseases and related disorders, indications, conditions, and syndromes associated with elevated cytokine expression in patients. Expression of various cytokines increases during inflammatory processes, including those secondary to circulatory shock, ischemia, reperfusion injury, and the like. TNF-α is a pleiotropic cytokine produced primarily by macrophages and other cell types. Other cytokines that increase during inflammatory processes, including those secondary to circulatory shock, ischemia, reperfusion injury, and the like, include IL-1 and IL-6. While cytokines such as TNF-α have beneficial effects in many instances, significantly elevated levels, such as those secondary to circulatory shock, ischemia, reperfusion injury, and the like, can have pathological effects. In one aspect, reperfusion of hypoxic or ischemic tissue, such as those secondary to circulatory shock, results in an inflammatory response, including elevated cytokine expression.

[0102] In one embodiment, the present invention is directed to methods of using one or more of the peptides of the present invention to reduce pro-inflammatory cytokine production and expression, including reducing pro-inflammatory cytokine production and expression secondary to circulatory shock, ischemia, reperfusion injury, etc. The reduction in the production and expression of pro-inflammatory cytokines, including but not limited to one or more of TNF-α, IL-1, and IL-6, preferably occurs within a short time following administration of a composition comprising one or more of the peptides of the present invention.

[0103] In related embodiments, the present invention is directed to methods of using one or more of the peptides and compositions of the present invention, including but not limited to, peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, to increase anti-inflammatory cytokine production and expression. The increased production and expression of anti-inflammatory cytokines, including but not limited to IL-10, occurs within a short time after administration of a composition comprising one or more of the peptides of the present invention.

[0104] 2.2 Dermatological Indications In yet another aspect, the peptides and compositions of the present invention, including but not limited to peptides that are MC1r agonists or partial agonists, can be used in the treatment of dermatological and cosmetic diseases, indications, conditions, and syndromes. In one aspect, the peptides and compositions of the present invention are MC1r agonists that stimulate melanocytes and related cells to increase the level of melanin in the skin. Increasing the level of melanin in the skin provides protection against ultraviolet radiation (UVR) and sunlight, including protection from cutaneous phototoxicity and photosensitivity caused by UVR, sun, and light.

[0105] In yet another aspect, the peptides and compositions of the present invention, including but not limited to peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be utilized for the prophylactic and / or therapeutic treatment of skin diseases, indications, conditions, and syndromes, such as acne vulgaris (commonly referred to as acne), atopic dermatitis (commonly referred to as atopic eczema or eczema), polymorphous light eruption, psoriasis, rosacea, seborrheic dermatitis, vitiligo, porphyria, porphyria cutanea tarda, erythropoietic protoporphyria, solar urticaria, urticaria pigmentosa, or xeroderma pigmentosum. In another aspect, the peptides, compositions, and methods of the present invention may be used to prevent, limit, or treat photosensitive or photoreactive viral infections, such as herpes simplex virus (commonly referred to as herpes labialis and genital herpes, depending on the site of infection), human papillomavirus, and varicella-zoster virus. In another aspect, the peptides, compositions, and methods of the present invention may be used to prevent, limit, or treat cancer of the skin, including use in precancerous conditions and in actinic keratosis, basal cell carcinoma, melanoma, or squamous cell carcinoma. In another aspect, the peptides, compositions, and methods of the present invention may be used to prevent or limit the adverse effects of various treatments, including phototherapy, such as photodynamic therapy. In yet another aspect, the peptides, compositions, and methods of the present invention may be used to prevent, limit, or treat photosensitive or photoreactive viral infections, such as herpes simplex virus (commonly referred to as herpes labialis and genital herpes, depending on the site of infection), human papillomavirus, and varicella-zoster virus. The peptides, compositions and methods may be utilized to induce tanning, reduce hair graying, or for similar and related purposes associated with increased melanin production.

[0106] 2.3 Cancer Certain cancers, such as mesothelioma, have been reported to be highly sensitive to the growth-promoting effects of cytokines and growth factors and may be treatable with peptides selective for MC1r. Canania, A., et al., "Autocrine inhibitory influences of α-melanocyte-stimulating hormone in malignant pleural mesothelioma," J. Leukoc. Biol. 75: 253-259 (2004). Cancers that may be so treated include other tumors that express MC1r, including, but not limited to, pleural mesothelioma, which is known to express MC1r and the receptor protein mRHA, as well as adenocarcinomas such as lung adenocarcinoma.

[0107] 2.4 Eye Diseases and Indications There are many ocular diseases, indications, conditions, and syndromes characterized by inflammation, including, but not limited to, elevated cytokine production, that can be treated with the peptides and compositions of the present invention, including, but not limited to, peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination of the foregoing. One example is dry eye disease, also known as dry eye syndrome or keratoconjunctivitis sicca, an ocular disorder that affects approximately 10-20% of the population. This disease progressively affects a higher proportion of the population as people age, and the majority of these patients are women. Furthermore, ocular irritation or symptoms and / or signs of dry eye as a condition are occasionally experienced under certain circumstances, such as prolonged visual tasks (e.g., computer work) in a dry environment, or the use of medications that cause eye dryness. In individuals suffering from dry eye, the protective tear layer that protects the ocular surface is typically compromised, resulting in insufficient or unhealthy production of one or more tear components. This can lead to the exposure of the ocular surface, ultimately promoting the drying and damage of surface cells.The signs and symptoms of dry eye include but are not limited to keratitis, conjunctiva and corneal staining, redness, blurred vision, shortened tear film breakup time, decreased tear production, tear volume and tear flow, increased conjunctival redness, excessive debris in tear film, dry eye, foreign body sensation in eye, burning eye, foreign body sensation in eye, excessive tear discharge, photophobia, stinging eye pain, refractive dysfunction, eye sensitivity and eye irritation.Patient may experience one or more of these symptoms.

[0108] There are many potential variables that can affect a patient's signs or symptoms of dry eye, including circulating hormone levels, various autoimmune diseases (e.g., Sjögren's syndrome and systemic lupus erythematosus), ocular surgery including PRK or LASIK, numerous medications, environmental conditions, visual tasks such as computer use, ocular fatigue, organic effects such as contact lens wear and corneal sensitivity, partial eyelid closure, surface irregularities (e.g., pterygium), and eyelid irregularities (e.g., ptosis, entropion / ectropion, pinguecula). Low humidity environments, such as dehydrating environments like sitting in a car with a defroster or living in a dry climate, can exacerbate or cause dry eye symptoms. Furthermore, visual tasks can exacerbate symptoms. Tasks that can significantly affect symptoms include prolonged TV or computer use, which reduces blinking rate.

[0109] Uveitis is an ocular disease involving inflammation of the middle layer of the eye, or uvea, and may also be understood to include any inflammatory process involving the interior of the eye. Uveitis includes anterior, intermediate, posterior, and panuveitis forms, with most uveitis cases located in the anterior segment and involving inflammation of the iris and anterior chamber. This condition may occur as a single episode and subside with appropriate treatment, or may be recurrent or chronic in nature. Symptoms include red eye, conjunctival hyperemia, pain, and decreased vision. Signs include dilated ciliary vessels, the presence of cells and flare in the anterior chamber, and keratin deposits on the posterior surface of the cornea. Intermediate uveitis is characterized by inflammation and nitric oxide. Posterior uveitis involves the presence of inflammatory cells in the retinal cavity, and posterior uveitis involves inflammation of the retina and choroid. Uveitis can be secondary to any of a number of diseases and disorders, including acute posterior multifocal discoid pigment epitheliopathy, ankylosing spondylitis, Behçet's disease, scattershot chorioretinopathy, 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's disease, or polyarteritis nodosa.

[0110] Other ocular inflammatory conditions for which one or more of the peptides of the present invention may be used to treat include corneal ulcers, corneal erosions, corneal abrasions, corneal degeneration, corneal perforation, corneal scarring, epithelial defects, keratoconjunctivitis, idiopathic uveitis, corneal transplants, age-related macular degeneration, diabetic eye pathologies, blepharitis, glaucoma, ocular hypertension, postoperative ocular pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascular membranes, vascular occlusive diseases, allergic eye diseases, tumors, retinal pigment epithelium, and the like. eye manifestations of systemic disease, hereditary eye conditions, eye tumors, increased intraocular pressure, herpetic infections, pterygium, persistent wounds to the surface of the eye, eye pain and inflammation after laser photorefractive keratectomy, thermal or chemical burns to the cornea, scleral wounds, keratoconus, or conjunctival wounds.

[0111] In one embodiment, the present invention is directed to methods of using one or more of the peptides of the present invention for the treatment of any of the aforementioned ocular diseases, indications, conditions, and syndromes. Such treatment may include treatment by eye drops, ointments, gels, rinses, implants, plugs, or other means and methods for delivering one or more of the peptides of the present invention to the surface of the eye, or alternatively by intravitreal injection or similar means that provide delivery to the vitreous humor, or alternatively by systemic administration, including oral, subcutaneous, or intravenous injection, to a patient who responds thereto.

[0112] For corneal transplants and similar ex vivo procedures, the present invention is directed to methods of using one or more of the peptides of the present invention for preservation of transplanted tissue, including, but not limited to, reducing pro-inflammatory cytokines, promoting IL-10 production, and improving endothelial cell survival. Thus, the present invention is directed to both methods of using one or more of the peptides of the present invention for preservation of transplanted tissue, and also to methods of treating patients immediately prior to, during, and after corneal transplantation, which treatment may include administration of eye drops and other topical administration of one or more of the peptides of the present invention.

[0113] 2.5 Ischemia and Related Indications In yet another aspect, the peptides and compositions of the present invention, including but not limited to peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be used in the treatment of ischemia and related diseases, indications, conditions, and syndromes. Ischemia includes any reduction or cessation of blood supply to any organ, tissue, cell, or part of the body, particularly where the reduction or cessation leads to or is thought to lead to ischemic damage to the organ, tissue, cell, or part of the body. An "ischemic episode" refers to any temporary or permanent period of ischemia. Ischemia can be the result of any narrowing or occlusion of the vasculature or of circulatory shock, such as hemorrhagic shock or hypovolemic shock. The reduction or lack of blood flow results in a reduction or lack of oxygen to the affected part of the body, which can also result in an increase in inflammatory disease-mediating chemicals, such as various cytokines and other substances. During certain surgical procedures, such as cardiac surgery and organ transplants, blood flow is temporarily stopped and then resumed (reperfusion), resulting in ischemia-reperfusion injury. During a heart attack, blood supply to the heart stops, resulting in ischemia, which can develop into an infarction. Current treatments for alleviating a heart attack involve reperfusion of the ischemic area of ​​the heart, such as by using thrombolytic drugs or coronary angioplasty.

[0114] The peptides and compositions of the present invention have particular application in preventing renal ischemic injury, including lung injury secondary to renal ischemia, preventing or limiting ischemic heart injury after myocardial infarction, and preventing or limiting ischemic brain injury after cardiovascular injury, including, but not limited to, myocardial infarction, stroke, etc. Administration of the compositions of the present invention to patients with cerebral ischemia or stroke, particularly those with concomitant hypotension, provides neuroprotection. The peptides and compositions of the present invention have further particular application in preventing or limiting ischemic organ damage in organ transplantation, including heart, kidney, liver, lung, pancreas, or small intestine transplantation. In one aspect, the pharmaceutical compositions of the present invention can be utilized for reperfusion of transplanted organs, which can be before, during, or after organ transplantation.

[0115] In one embodiment, the present invention is directed to a method of using one or more of the peptides of the present invention to protect the heart, brain, or other organs of a patient from damage caused by ischemia, wherein the protective effect against ischemia occurs immediately or within a short period of time after administration of a composition comprising one or more of the peptides of the present invention.

[0116] Ischemia can result from any of a variety of diseases or conditions, and in one embodiment, the invention is directed to a method of using one or more peptides of the invention to protect a patient's organs from damage resulting from ischemia, where the ischemia is caused by a disease or condition. Examples of such diseases or conditions include, but are not limited to, atherosclerotic disease, e.g., atheroma with thrombosis, embolism from blood vessels from the heart or any organ, vasospasm, hypotension due to heart disease, hypotension due to systemic disease, including infection or allergic reaction, or hypotension resulting from the administration, ingestion, or other exposure to one or more toxic compounds or drugs. Ischemia can also be secondary, and in another embodiment, the invention is directed to a method of using one or more peptides of the invention to protect a patient's organs from damage resulting from secondary ischemia. Such secondary ischemia may be secondary to diseases or conditions such as diabetes, hyperlipidemia, hyperlipoproteinemia, dyslipidemia also known as thromboangiopathy obliterans, Buerger's disease, Takayasu's arteritis, temporal arteritis, Kawasaki disease also known as lymph node syndrome, mucocutaneous nodular disease, infantile polyarteritis, cardiovascular syphilis, and various connective tissue diseases and disorders.

[0117] In yet another aspect, the peptides and compositions of the present invention, including but not limited to peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be used in the treatment of ischemia-reperfusion injury and related diseases, indications, conditions, and syndromes. While restoration of blood flow after ischemia is essential to preserve functional tissue, reperfusion itself is known to be harmful to tissue. Both ischemia and reperfusion are known to be important contributors to tissue necrosis. Several mechanisms are believed to play a causative role in the generation of tissue damage associated with ischemia-reperfusion injury. Certain of the peptides and compositions of the present invention have particular application in preventing or limiting the severity of renal reperfusion injury, including lung injury secondary to renal reperfusion, preventing or limiting reperfusion cardiac injury after myocardial infarction, and preventing or limiting reperfusion brain injury after cardiovascular injuries, including, but not limited to, myocardial infarction, stroke, and the like. The present invention has further particular application in preventing or limiting reperfusion organ injury in organ transplantation, including heart, kidney, liver, lung, pancreas, or small intestine transplantation. In one aspect, the pharmaceutical compositions of the present invention may be utilized for perfusion of the transplanted organ, which may be before, during, or after organ transplantation.

[0118] In one embodiment, the present invention is directed to a method of using one or more peptides of the present invention to protect a patient's heart, brain, or other organ from damage caused by ischemia-reperfusion injury, including damage caused by or during reperfusion, wherein the protective effect from ischemia-reperfusion injury occurs immediately or within a short time after administration of a composition comprising one or more peptides of the present invention.

[0119] In yet another aspect, peptides and compositions of the invention, including but not limited to peptides that are MC1r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be used in the treatment of circulatory shock and related diseases, indications, conditions, and syndromes in patients. The invention provides peptides, compositions for use, and methods for treating or preventing shock, including hemorrhagic shock, in patients, comprising administering a composition comprising one or more of the peptides of the invention to a patient diagnosed with blood loss. Blood loss may, but need not, be measured as a percentage of the subject's blood volume, such as, for example, blood loss of more than about 15% of the total blood volume or more than 20%, 25%, 30%, 35%, 40%, or 50% of the subject's total volume. Alternatively, blood loss may, but need not, be measured in terms of a decrease in blood volume of any amount sufficient to cause hemorrhagic shock in a particular subject, such as, for example, a loss of about 750 mL, 1000 mL, about 1500 mL, or about 2000 mL or more in a human subject. Blood loss may also be measured in terms of a decrease in systolic blood pressure, such as, for example, a decrease in systolic blood pressure of about 20 mmHg, 30 mmHg, 40 mmHg, 50 mmHg, 60 mmHg, 70 mmHg, 80 mmHg, 90 mmHg, or 100 mmHg or more than 100 mmHg below the subject's normal systolic blood pressure. In certain embodiments, the subject is undergoing or has undergone a medical procedure, such as, but not limited to, surgery, transfusion, or childbirth. In other specific embodiments, the subject has suffered a traumatic injury, such as, but not limited to, that resulting from a car accident, industrial accident, or gunshot wound.

[0120] In further embodiments of the present invention, the compositions and methods are used to treat cardiogenic shock, hypovolemic shock, and vasodilatory shock, each of which may be at any stage of shock. In one specific embodiment of the present invention, the method is used to treat cardiogenic shock. Cardiogenic shock, generally speaking, is low blood flow or perfusion caused by cardiac dysfunction, in which the heart does not pump adequate blood. Causes may include any condition that interferes with ventricular filling or emptying, such as, but not limited to, embolism, ischemia, regurgitation, and valvular dysfunction. In another specific embodiment of the present invention, the method is used to treat vasodilatory shock. Vasodilatory shock is caused by severe venous or arteriolar dilation, resulting in insufficient blood flow. Several known causes contribute to vasodilatory shock, including, but not limited to, brain trauma, drug or toxin toxicity, anaphylaxis, liver failure, bacteremia, and sepsis. In another more specific embodiment of the present invention, the method is used to treat shock resulting from sepsis or bacteremia. In even more specific embodiments, the compositions and methods are used to treat septic or bacteremic shock, referred to as stage I, II, or III shock. In yet another embodiment, the compositions and methods of the present invention are used to treat hypovolemic shock. Hypovolemic shock generally results in a decrease in intravascular volume, and the decrease in intravascular volume can be relative or absolute. Bleeding from conditions such as, but not limited to, ulcers, gastrointestinal injuries, trauma, accidents, surgery, and aneurysms can cause hypovolemic shock; however, other fluid losses can also cause hypovolemic shock. For example, renal fluid loss, intravascular fluid loss, and loss of water or other ascites fluid can contribute to hypovolemic shock. In a specific embodiment of the present invention, compositions and methods comprising administration of one or more peptides of the present invention are used to treat hypovolemic shock. In an even more specific embodiment, the compositions and methods are used to treat stage I, stage II, or stage III hypovolemic shock.

[0121] In one embodiment, the present invention is directed to a method of using one or more peptides of the present invention to protect the heart, brain, or other organs of a patient from damage caused by circulatory shock, wherein the protective effect from circulatory shock occurs immediately or within a short time after administration of a composition comprising one or more peptides of the present invention, preferably within at least about 40 minutes after administration.

[0122] 2.6 Indications for MC4r responsiveness In another aspect, the peptides and compositions of the present invention, including but not limited to peptides that are MC4r agonists or partial agonists or MC3r agonists or partial agonists or any combination thereof, are useful for treating energy homeostasis and metabolism-related (such as diabetes, particularly type 2 diabetes; dyslipidemia; fatty liver; gout; hypercholesterolemia; hypertriglyceridemia; hyperuricemia; impaired glucose tolerance; fasting hyperglycemia; insulin resistance syndrome; and metabolic syndrome), eating-related (such as bulimia; overeating; increased appetite; and psychogenic overeating) and / or energy balance and weight-related diseases, disorders and / or conditions. may be used in treating conditions, more particularly diseases, disorders and / or conditions responsive to modulation of MC4r function, more particularly the activation of MC4r, i.e., diseases, disorders and / or conditions that would benefit from agonism at MC4r (including full or partial agonism), including diseases, disorders and / or conditions characterized by overweight and / or overeating, or diseases, disorders and / or conditions responsive to modulation of MC3r function, more particularly the activation of MC3r, i.e., diseases, disorders and / or conditions that would benefit from agonism at MC3r (including full or partial agonism) or modulation of both MC4r and MC3r function. In one aspect, the compounds of the present invention are utilized to treat conditions associated with various expression or receptor inherited disorders, such as pro-opiomelanocortin deficiency due to mutations in the POMC gene (POMC heterozygous deficiency obesity), Prader-Willi syndrome, obesity due to MC4r deficiency, leptin receptor deficiency obesity, leptin deficiency obesity including congenital leptin deficiency, Bardet-Biedl syndrome, Alström syndrome, and various other diseases, conditions, gene defects, metabolic disorders, and syndromes.

[0123] Such peptides are particularly believed to be useful for the treatment of excess weight, including obesity and overweight (by promoting weight loss, maintaining weight loss and / or preventing weight gain, including weight gain secondary to drug-induced weight gain or smoking cessation), and weight-related diseases, disorders and / or conditions associated with obesity and / or overweight, such as insulin resistance; impaired glucose tolerance; type 2 diabetes; metabolic syndrome; dyslipidemia (including hyperlipidemia); hypertension; heart disorders (e.g., coronary heart disease, myocardial infarction); cardiovascular disorders; non-alcoholic fatty liver disease (including non-alcoholic steatohepatitis); joint disorders (including secondary osteoarthritis); gastroesophageal reflux; sleep apnea; atherosclerosis; stroke; macrovascular and microvascular disease; steatosis (e.g., in the liver); gallstones; and gallbladder disorders.

[0124] MC4r is part of the leptin-melanocortin pathway or pro-opiomelanocortin (POMC)-MC4r pathway. Members of this pathway include a wide variety of proteins, including α-MSH, POMC, leptin, and the leptin receptor. Certain diseases, conditions, and syndromes are associated with the POMC-MC4r pathway or arise from mutations and variations, including genetic defects, in one or more members of the POMC-MC4r pathway. The compounds of the present invention may be useful in treating diseases, conditions, and syndromes associated with or arise from mutations and variations, including genetic defects, in one or more members of the POMC-MC4r pathway, as described herein below.

[0125] The hypothalamic POMC-MC4r pathway is part of a regulatory system that regulates feeding behavior, appetite, and body weight. Numerous diseases, conditions, and syndromes have been described that are associated with disruption of the hypothalamic POMC-MC4r pathway, which are believed to result from genetic defects or disruptions, including genetic defects or disruptions in the POMC-MC4r pathway. For example, Prader-Willi syndrome manifests as marked hyperphagia and severe obesity and may include other characteristics and symptoms, such as learning disabilities, neurological dysfunction, hypogonadism, short stature, and developmental and cognitive delays. The compounds of the present invention may be useful for treating Prader-Willi syndrome and other diseases, conditions, and syndromes involving genetic defects or disruptions in the POMC-MC4r pathway, as described later herein.

[0126] Thus, the compounds of the present invention can be used and are indicated for the treatment of obesity and hyperphagia associated with POMC deficiency caused by homozygous or compound heterozygous loss-of-function mutations in the POMC gene located at position 23.3 on chromosome 2. Mutations in the POMC gene that cause complete loss or significantly reduce production of POMC polypeptide lead to the absence or reduced production of α-MSH. This loss of endogenous α-MSH results in significantly reduced MC4r activity, leading to hyperphagia and obesity. The compounds of the present invention can be used as supplemental MC4r agonist therapy in patients with low or no endogenous α-MSH.

[0127] For various diseases, conditions, or syndromes associated with disruption of the hypothalamic POMC-MC4r pathway, various genetic and genotyping tests can be used as part of a prospective patient's diagnosis to determine the suitability of such prospective patient for use with the compounds of the present invention. As a non-limiting example for Prader-Willi syndrome, genetic tests such as DNA-based methylation testing can be used to confirm loss of active genes in specific portions of chromosome 15, the 15q11-q13 region, specifically deletion of at least the 15q11-q13 region of paternal chromosome 15. Similarly, POMC deficiency can be diagnosed by loss-of-function mutations in the POMC gene. Therefore, treatment with the compounds of the present invention can include various diagnostic and genetic tests to confirm the presence of loss-of-function mutations in the POMC-MC4r pathway or other mutations, including, but not limited to, loss-of-function mutations for Prader-Willi syndrome affecting the 15q11-q13 region, loss-of-function mutations in the POMC gene, leptin gene, leptin receptor gene, and various other genes in the POMC-MC4r pathway.

[0128] In yet another aspect, peptides and compositions of the present invention, including but not limited to peptides that are MC4r agonists or partial agonists, can be used to treat sexual dysfunction, including both male erectile dysfunction and female sexual dysfunction. Female sexual dysfunction includes, but is not limited to, hypoactive sexual desire disorder. In one specific embodiment, the peptides, compositions, and methods of the present invention are used in male patients to improve erectile function, including but not limited to, improving erectile function to enable vaginal intercourse. In another specific embodiment, the peptides, compositions, and methods of the present invention are used to treat female sexual dysfunction, including but not limited to, improving sexual arousal success, desire success, and levels of sexual arousal and desire. For female sexual dysfunction, including hypoactive sexual desire disorder, endpoints can be determined by any of a number of validated instruments, including, but not limited to, the Female Sexual Distress Scale, Female Sexual Encounter Profile, Female Sexual Function Index, and Global Assessment Questionnaire. Patients treated for female sexual dysfunction can be premenopausal or postmenopausal women.

[0129] In yet another embodiment, the peptides and compositions of the present invention, including but not limited to peptides that are MC4r agonists or partial agonists, can be used to inhibit or reduce alcohol consumption, treat or prevent alcoholism, treat or prevent alcohol abuse, or treat or prevent alcohol-related disorders. In another related embodiment, one or more of the peptides can be used to inhibit consumption of drugs of abuse, reduce consumption of drugs of abuse, treat or prevent drug abuse, or treat or prevent drug abuse-related disorders. Drugs of abuse are generally controlled substances. These include controlled naturally occurring drugs such as heroin, morphine, opium, cocaine, marijuana, etc., as well as synthetic drugs such as Vicodin®, Lortab®, Lorcet®, Percocet®, Percodan®, Tylox®, Hydrocodone, OxyContin®, methadone, tramadol, various methamphetamines and other tranquilizers, stimulants, or drugs of abuse. These include known sedatives as well as drugs with no established pharmaceutical utility, such as ecstasy, LSD, or PCP.

[0130] In yet another aspect, the peptides and compositions of the present invention, including, but not limited to, MC4r antagonist or optionally inverse agonist peptides at MC4r, including MC4r antagonist or inverse agonist peptides, which may be agonists, partial agonists, antagonists, or inverse agonists at one or more of MC1r, MC2r, MC3r, and MC5r, can be used in the treatment of various body weight disorders, including cachexia, sarcopenia, and wasting syndromes or diseases, and for the treatment of inflammatory and immune disorders. Body weight disorders include one or more "wasting" disorders (e.g., wasting syndromes, cachexia, sarcopenia) that cause undesirable and unhealthy weight loss or loss of body cell mass. In elderly individuals and cancer and AIDS patients, wasting disorders can result in undesirable weight loss, including both fat and fat-free compartments. Wasting disorders can be the result of inadequate food intake and / or metabolic changes associated with disease and / or the aging process. Cancer and AIDS patients, as well as patients after extensive surgery or those with chronic infectious or immunological disorders, hyperthyroidism, Crohn's disease, psychogenic disorders, chronic heart failure, or other severe trauma, often suffer from wasting diseases. Wasting diseases, sometimes called cachexia, are generally recognized as metabolic disorders and sometimes as eating disorders. Cachexia can also be characterized by hypermetabolism and hypercatabolism. Sarcopenia, another such disorder that can affect elderly people, is generally characterized by loss of muscle mass. Such end-stage wasting diseases can occur in individuals suffering from either cachexia or sarcopenia.

[0131] 2.7 Nuclear Medicine and Drug Delivery Applications In yet another aspect, peptides and compositions of the invention, including but not limited to peptides that are MC1r agonists, partial agonists, or antagonists, can be used in targeted imaging and cytotoxic therapy for certain cancers, such as melanoma, and other indications, in patients in need thereof. The peptides, compositions, and methods of the invention can be used to image melanoma and other cancers or diseases or conditions characterized in part by relatively high expression of MC1r, such as by diagnostic imaging using a radionuclide in combination with the peptides of the invention. For diagnostic imaging, the peptides of the invention are typically conjugated to a radionuclide by the use of a linker, e.g., a crosslinking agent, that couples the peptide to the radionuclide. The radionuclide is preferably a gamma emitter that can be imaged using a gamma detector or camera, such as single-photon emission computed tomography, or a positron emitter that can be imaged using positron emission tomography. Gamma emitters that can be used in this way include, inter alia, 99m Tc, 111 In, 123 I and 67 Examples of positron emitters that can be used in this way include Ga. 11 C. 13 N, 15 O and 18 F is one example.

[0132] In a related aspect, the peptides, compositions, and methods of the invention can be used for cytotoxic therapy of melanoma, other cancers, or diseases or conditions characterized in part by relatively high expression of MC1r, such as by utilizing chemotherapeutic or radiotherapeutic agents, including toxins, in combination with the peptides of the invention. Chemotherapeutic agents include any antineoplastic or chemical agent, such as alkylating agents, antimetabolites, anthracyclines, plant alkaloids, topoisomerase inhibitors, and other antitumor agents. Non-limiting examples of alkylating agents include cisplatin, carboplatin, oxaliplatin, mechlorethamine, cyclophosphamide, chlorambucil, and ifostamide; examples of antimetabolites include azathioprine and mercaptopurine; examples of anthracyclines include daunorubicin, doxorubicin, epirubicin, idarubicin, valrubicin, and mitoxantrone; examples of plant alkaloids include vinca alkaloids, such as vincristine, vinblastine, vinorelbine, and vindesine, and taxanes, such as paclitaxel and docetaxel. Examples of topoisomerase inhibitors include camptothecins, such as irinotecan and topotecan, and type II topoisomerase inhibitors, such as amsacrine, etoposide, etoposide phosphate, and teniposide. However, any agent suitable for use in targeted cytotoxic therapy may be so used. Non-limiting examples of radiotherapeutic agents that may be so used include, inter alia, 131 I, 125 I, 211 At, 186 Re, 188 Re, 90 Y, 153 Sm, 212 Bi and 32 Examples include P.

[0133] For example, instead of non-radioactive isotopes, among others 11 C. 13 N, 15Diagnostic imaging or cytotoxic therapeutic agents can be coupled to the peptides of the invention, such as by use of O; can be directly linked to the peptides of the invention, such as by halogenation or other direct complexation methods; or can be indirectly linked to the peptides of the invention, such as by conjugation reactions through linker or chelating units. Linker units are well known in the art and include, but are not limited to, chemically linked conjugates comprising at least one disulfide bond, thioether bond, or covalent bond between free reactive groups. Representative cross-linking and conjugation reagents are disclosed, inter alia, in U.S. Pat. Nos. 7,169,603, 7,820,164, and 5,443,816 and U.S. Patent Application Publication No. 2009 / 0297444, which are incorporated herein by reference.

[0134] 3.0 Combination Therapies for Certain Indications The peptides, compositions, and methods of the present invention can be used to treat any of the aforementioned diseases, indications, conditions, or syndromes, or any disease, indication, condition, or syndrome mediated or responsive to MC1r, by administration in combination with one or more other pharmaceutically active compounds. Such combined administration can be via a single dosage form containing both the peptide of the present invention and one or more other pharmaceutically active compounds, including tablets, capsules, sprays, inhalation powders, liquids for injection, and the like. Alternatively, combined administration can be via administration of two different dosage forms, one containing the peptide of the present invention and the other containing another pharmaceutically active compound. In this example, the dosage forms can be the same or different. The term "co-administration" indicates that each of at least two compounds in a combination therapy is administered during a time frame in which their respective periods of biological activity or effect overlap. Thus, this term includes sequential administration as well as simultaneous administration of compounds, where one compound is one or more peptides of the present invention. When multiple compounds are co-administered, the routes of administration of the two or more compounds need not be the same. Without intending to be limiting on combination therapy, the following are illustrative of certain combination therapies that may be used.

[0135] 3.1 Combination therapy with anti-inflammatory agents For the treatment of inflammation-related diseases, indications, conditions, and syndromes, the peptides of the present invention can be used in combination therapy, including by co-administration with one or more anti-inflammatory agents. One class of anti-inflammatory agents is the glucocorticoids, including, but not limited to, cortisone acetate, hydrocortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, prednisone, fludrocortisone acetate, deoxycorticosterone acetate, and aldosterone. Other anti-inflammatory agents that may be used in combination therapy, including by co-administration, include aspirin, nonsteroidal anti-inflammatory drugs (NSAIDs) (such as ibuprofen and naproxen), TNF-α inhibitors (such as tenidap and rapamycin or its derivatives) or TNF-α antagonists (e.g., infliximab, OR1384), cyclooxygenase inhibitors (i.e., COX-1 and / or COX-2 inhibitors, such as naproxen® or Celebrex®), CTLA4-Ig agonists / antagonists, CD40 ligand antagonists, IMPDH inhibitors such as mycophenolate (Cellcept®), integrin antagonists, alpha-4 beta-7 integrin antagonists, cell adhesion inhibitors, interferon gamma antagonists, INF-α inhibitors, and the like. CAM-1, prostaglandin synthesis inhibitors, budesonide, clofazimine, p38 mitogen-activated protein kinase inhibitors, protein tyrosine kinase (PTK) inhibitors, IKK inhibitors, therapeutic agents for the treatment of irritable bowel syndrome (e.g., Zelmac® and Maxi-K® openers, such as those disclosed in U.S. Pat. No. 6,184,231) or other NF-κB inhibitors, such as corticosteroids, calphostin, CSAIDs, 4-substituted imidazo[1,2-α]quinoxalines such as those disclosed in U.S. Pat. No. 4,200,750; interleukin-10, salicylates, nitrate oxides, and other immunosuppressants; and nuclear transfer inhibitors, such as deoxyspergualin (DSG).

[0136] 3.2 Combination therapy with phosphodiesterase inhibitors For certain applications and indications, it is desirable to enhance the production and maintain levels of cyclic adenosine 3',5'-monophosphate (cAMP), a nucleotide messenger associated with inflammatory cell activity. The peptides of the present invention can be co-administered with compounds or substances that enhance intracellular levels of cAMP and inhibit the degradation of cAMP. cAMP is hydrolyzed to its inactive form by phosphodiesterases (PDEs); compounds or substances that inhibit PDEs can maintain and / or increase available cAMP. The class of compounds known as PDE inhibitors has been widely studied for use in the treatment of inflammatory diseases, such as asthma, COPD, and acute respiratory distress syndrome. Inhibitors of PDE types 1, 2, 3, 4, 7, 8, 10, or 11 are preferred; in one embodiment, these include cAMP-PDE inhibitors, which are selective PDE type 4 inhibitors or inhibitors with selectivity for specific types of PDE4 isozymes, such as rolipram, cilomilast, ibudilast, and piclamilast.

[0137] 3.3 Combination Therapies for Ophthalmic Indications For ophthalmic indications, ophthalmic dosage forms may contain one or more active ingredients in addition to one or more of the peptides of the invention, such as, for example, an artificial tear component, a topical corticosteroid, a nonsteroidal anti-inflammatory drug, or a calcineurin inhibitor, e.g., cyclosporine-A ophthalmic emulsion (Restasis® - Allergan). Co-administration can also include administration of one or more additional compounds given separately from the peptides of the invention, such as separate administration of an ophthalmic dosage form containing an artificial tear component, a topical corticosteroid, a nonsteroidal anti-inflammatory drug, a calcineurin inhibitor, e.g., cyclosporine-A, or a combination of any of the foregoing.

[0138] Combination ophthalmic solutions may be used, including solutions containing multiple active pharmaceutical ingredients. In one embodiment, a nonsteroidal anti-inflammatory drug (NSAID) is used in combination with the peptide of the present invention. Suitable NSAIDs for use in combination ophthalmic solutions include propionic acid compounds such as naproxen, flurbiprofen, oxaprozin, ibuprofen, ketoprofen, fenoprofen; ketorolac tromethamine, etc.; acetic acid derivatives such as sulindac, indomethacin, and etodolac; phenylacetic acids such as diclofenac, bromfenac, and suprofen; arylacetic acid prodrugs such as nepafenac and amfenac; salicylates such as aspirin, salsalate, diflunisal, choline magnesium trisalicylate, etc.; para-aminophenol derivatives such as acetaminophen. and the like; naphthyl alkanones, such as nabumetone; enolic acid derivatives, such as piroxicam and meloxicam; femanates, such as mefenamic acid, meclofenamic acid, and flufenamic acid; pyrroleacetic acids, such as tolmetin; and pyrazolones, such as phenylbutazone; and cyclooxygenase (COX)-2 selective inhibitors, such as celecoxib, valdecoxib, parecoxib, etoricoxib, and rualicoxib, which inhibit the cyclooxygenase (COX)-1 and / or -2 enzymes, their esters, and pharmaceutically acceptable salts thereof, including, but not limited to, vasoconstrictors, antiallergics, anti-infectives, steroids, anesthetics, anti-inflammatory agents, analgesics, drug It may further comprise other active ingredients, including, but not limited to, dry eye treatments (e.g., secretagogues, mucus mimetics, polymers, lipids, antioxidants), or may be administered in conjunction with (concurrently or sequentially with) pharmaceutical compositions containing other active ingredients, including, but not limited to, vasoconstrictors, antiallergic agents, anti-infectives, steroids, anesthetics, anti-inflammatory agents, analgesics, dry eye treatments (e.g., secretagogues, mucus mimetics, polymers, lipids, antioxidants), etc.

[0139] 3.4 Concomitant Therapy in Shock-Related Indications The methods of treating or preventing circulatory shock of the present invention also involve co-administering one or more substances to a subject in addition to one or more peptides of the present invention. For example, one or more peptides of the present invention can be co-administered with androstentriol, androstenediol or derivatives thereof, various vasopressin agonists, or other pharmaceutically active substances, such as other alpha adrenergic agonists, alpha 2 adrenergic agonists, beta adrenergic agonists, or beta 2 adrenergic agonists, including, but not limited to, catecholamines or epinephrine, norepinephrine, dopamine, isoproterenol, vasopressin, and dobutamine. Alternatively, one or more peptides of the present invention can be co-administered with a fluid or other substance capable of alleviating, attenuating, preventing, or eliminating symptoms in a subject suffering from, experiencing, or at risk of suffering from hypovolemic shock, vasodilatory shock, or cardiogenic shock. The type of fluid that can be co-administered with one or more peptides of the present invention should be specific to the circumstances of a particular subject suffering from, experiencing, or at risk of suffering from shock. For example, fluids that can be co-administered with one or more peptides of the present invention include, but are not limited to, salt solutions (e.g., sodium chloride and sodium bicarbonate), as well as whole blood, synthetic blood substitutes, plasma, serum, serum albumin, and colloid solutions. Colloid solutions include, but are not limited to, solutions containing hetastarch, albumin, or plasma. In a specific embodiment of the present invention, a fluid such as one or more of a salt solution, colloid solution, whole blood, synthetic blood substitute, plasma, or serum is co-administered with one or more peptides of the present invention to a patient suffering from or experiencing hypovolemic shock, such as hemorrhagic shock.

[0140] 3.5 Combination Therapies for Obesity and Related Metabolic Syndrome One or more peptides of the present invention may be combined with one or more other pharmacologically active agents useful in the treatment of various weight- and eating-related disorders, such as obesity and / or excess weight, in particular other anti-obesity agents that affect energy expenditure, glycolysis, gluconeogenesis, glucogenolysis, lipolysis, lipogenesis, fat absorption, fat storage, fat elimination, hunger and / or satiety and / or craving mechanisms, appetite / motivation, food intake, or gastrointestinal motility. Agents that reduce energy intake include, in part, various pharmacological substances called anorectic agents, which are used as adjuncts to behavioral therapy in weight loss programs.

[0141] Generally, the total dosage of the obesity control agents or drugs described below, when used in combination with one or more peptides of the present invention, can range from 0.01 to 3,000 mg / day, preferably about 0.1 to 50 mg / day, and more preferably about 0.1 to 10 mg / day, in a single dose or in 2 to 4 divided doses. However, the exact dosage will be determined by the attending clinician and will depend on factors such as the potency of the compound administered, and the age, weight, condition, and response of the patient.

[0142] One or more peptides of the present invention may be combined with one or more other pharmacologically active substances that are useful in the treatment of diabetes, such as other antidiabetic agents.

[0143] One or more peptides of the present invention may also or alternatively be used to treat conditions such as: insulin resistance; impaired glucose tolerance; type 2 diabetes; metabolic syndrome; dyslipidemia (including hyperlipidemia); hypertension; cardiac disorders (e.g., coronary heart disease, myocardial infarction); cardiovascular disorders; non-alcoholic fatty liver disease (e.g., The compound may further be combined with one or more other pharmacologically active substances that are useful in the treatment of diseases, disorders and / or conditions associated with obesity and / or excess weight, such as: obesity, urinary tract infections (including non-alcoholic steatohepatitis); joint disorders (including secondary osteoarthritis); gastroesophageal reflux; sleep apnea; atherosclerosis; stroke; macrovascular and microvascular disease; steatosis (e.g., in the liver); gallstones; and gallbladder disorders.

[0144] According to a further aspect of the present invention, a mammal, such as a human, in need of such therapeutic treatment is administered one or more of the following agents, optionally together with a pharmaceutically acceptable carrier: - insulin and insulin analogues; - insulin secretagogues, including sulfonylureas (e.g. glipizide) and dietary glucose control agents (sometimes called "short-acting secretagogues"), such as meglitinides (e.g. repaglinide and nateglinide); - Medications that improve incretin action, such as dipeptidyl peptidase IV (DPP-4) inhibitors (e.g., vildagliptin, saxagliptin, and sitagliptin) and glucagon-like peptide-1 (GLP-1) agonists (e.g., exenatide); - insulin sensitizers, including peroxisome proliferator-activated receptor gamma (PPARγ) agonists such as thiazolidinediones (e.g., pioglitazone and rosiglitazone) and agents with any combination of PPAR alpha, gamma, and delta activity; - drugs that regulate hepatic glucose balance, such as biguanides (e.g. metformin), fructose 1,6-bisphosphatase inhibitors, glycogen phosphorylase inhibitors, glycogen synthase kinase inhibitors and glucokinase activators; - Medications designed to reduce / delay the absorption of glucose from the intestine, such as alpha-glucosidase inhibitors (e.g., miglitol and acarbose); - Medications that antagonize the action of glucagon or reduce its secretion, such as amylin analogues (e.g., pramlintide); - Medications that prevent glucose reabsorption by the kidney, such as sodium-dependent glucose transporter 2 (SGLT-2) inhibitors (e.g., dapagliflozin); - Medications designed to treat the complications of long-term hyperglycemia, such as aldose reductase inhibitors (e.g., epalrestat and ranirestat); and medications used to treat complications associated with microangiopathy; - anti-dyslipidemic agents, such as HMG-CoA reductase inhibitors (statins, e.g., rosuvastatin) and other cholesterol-lowering agents; PPARα agonists (fibrates, e.g., gemfibrozil and fenofibrate); bile acid sequestrants (e.g., cholestyramine); cholesterol absorption inhibitors (e.g., plant sterols (i.e., phytosterols), synthesis inhibitors); cholesteryl ester transfer protein (CETP) inhibitors; inhibitors of the ileal bile acid transport system (IBAT inhibitors); bile acid-binding resins; nicotinic acid (niacin) and its analogs; antioxidants, such as probucol; and omega-3 fatty acids; - antihypertensives, including adrenergic receptor antagonists, such as beta-blockers (e.g., atenolol), alpha-blockers (e.g., doxazosin), and mixed alpha / beta-blockers (e.g., labetalol); adrenergic receptor agonists, including alpha-2 agonists (e.g., clonidine); angiotensin-converting enzyme (ACE) inhibitors (e.g., lisinopril), calcium channel blockers, such as dihydropridines (e.g., nifedipine), phenylalkylamines (e.g., verapamil), and benzothiazepines (e.g., diltiazem); angiotensin II receptor antagonists (e.g., candesartan); aldosterone receptor antagonists (e.g., eplerenone); centrally acting adrenergic drugs, such as central alpha agonists (e.g., clonidine); and diuretics (e.g., furosemide); - Hemostatic regulators, including antithrombotic agents, such as activators of fibrinolysis; thrombin antagonists; factor VIIa inhibitors; anticoagulants, such as vitamin K antagonists (e.g. warfarin), heparin and its low molecular weight analogues, factor Xa inhibitors and direct thrombin inhibitors. inhibitors (e.g., argatroban); antiplatelet agents, such as cyclooxygenase inhibitors (e.g., aspirin), adenosine diphosphate (ADP) receptor inhibitors (e.g., clopidogrel), phosphodiesterase inhibitors (e.g., cilostazol), glycoprotein IIB / IIA inhibitors (e.g., tirofiban), and adenosine reuptake inhibitors (e.g., dipyridamole); - anti-obesity agents, such as appetite suppressants (e.g. ephedrine), including noradrenergic agents (e.g. phentermine) and serotonergic agents (e.g. sibutramine), pancreatic lipase inhibitors (e.g. orlistat), microsomal transfer protein (MTP) modulators, diacylglycerol acyltransferase (DGAT) inhibitors and cannabinoid (CB1) receptor antagonists (e.g. rimonabant); Feeding behavior modulators, such as orexin receptor modulators and melanin-concentrating hormone (MCH) modulators; - Glucagon-like peptide-1 (GLP-1) receptor modulators; - Neuropeptide Y (NPY) / NPY receptor modulators; - Pyruvate dehydrogenase kinase (PDK) regulators; - Serotonin receptor modulators; - Leptin / leptin receptor modulators; - ghrelin / ghrelin receptor modulators; or monoamine transport modulators, such as selective serotonin reuptake inhibitors (SSRIs) (e.g. fluoxetine), noradrenaline reuptake inhibitors (NARIs), noradrenaline-serotonin reuptake inhibitors (SNRIs), triple monoamine reuptake blockers (e.g. tesofensine) and monoamine oxidase inhibitors (MAOIs) (e.g. toloxatone and amiflamine), or Pharmaceutically acceptable salts, solvates, solvates of such salts or prodrugs thereof The present invention provides a combination treatment comprising administering a pharmacologically effective amount of a peptide according to the present invention or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable diluent or carrier, in simultaneous, sequential or separate administration with one or more of the following:

[0145] According to a further aspect of the present invention there is provided a combination treatment comprising administering a pharmacologically effective amount of a compound according to the invention or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable carrier, in simultaneous, sequential or separate administration of a very low calorie diet (VLCD) or a low calorie diet (LCD).

[0146] Additional aspects of the present invention as disclosed in WO 2016 / 168388, "Therapies for Obesity, Diabetes and Related Indications," which is incorporated herein by reference. According to a further possible embodiment, one or more peptides of the invention, and preferably peptides that are MC4r agonists, may be administered in conjunction with a GLP-1 receptor agonist. Thus, the invention herein includes pharmaceutical compositions for subcutaneous administration in the treatment of obesity or for inducing weight loss, including on a per dose basis: a peptide of the invention that is an MC4r agonist in an amount sufficient to induce at least minimal weight loss when administered as monotherapy without a GLP-1 receptor agonist; and An amount of a GLP-1 receptor agonist sufficient to induce glycemic control but not weight loss when administered as monotherapy without an MC4r agonist, wherein the pharmaceutical composition preferably has a synergistic anti-obesity effect.

[0147] In a related aspect, the present invention provides a method for treating a vascular endothelial disorder comprising administering to a subject a peptide of the present invention comprising: (a) a peptide of the present invention that is an MC4r agonist in an amount sufficient to induce at least minimal weight loss when administered as monotherapy without a GLP-1 receptor agonist; and (b) a peptide of the present invention that is an MC4r agonist in an amount sufficient to induce glycemic control but not weight loss when administered as monotherapy without a GLP-1 receptor agonist. A method of treating a patient suffering from obesity, diabetes or metabolic syndrome is provided, comprising administering to the patient a P-1 receptor agonist, which preferably induces a synergistic effect on the treatment of obesity.

[0148] In another aspect, the present invention provides a method of reducing side effects associated with a therapeutic agent for the treatment of obesity, diabetes, or metabolic syndrome in a patient, the method comprising: administration of a peptide of the invention that is an MC4r agonist in an amount such that when administered as monotherapy without a GLP-1 receptor agonist, the amount of the MC4r agonist peptide administered is not sufficient to initiate a desired pharmacological response in treating at least one condition from the group consisting of obesity, diabetes, and metabolic syndrome in a patient when administered as monotherapy; and Administration of a GLP-1 receptor agonist in an amount such that when administered as monotherapy without an MC4r agonist, the amount of the administered GLP-1 receptor agonist is not sufficient to initiate a desired pharmacological response in treating at least one condition from the group consisting of obesity, diabetes, and metabolic syndrome in a patient when administered as monotherapy. Includes; The amount of the MC4r agonist and the amount of the GLP-1 receptor agonist together are effective to initiate a desired pharmacological response to treat at least one condition from the group consisting of obesity, diabetes, and metabolic syndrome in a patient, thereby reducing side effects in treating at least one of obesity, diabetes, or metabolic syndrome in a patient.

[0149] 3.6 Combination Therapy for Sexual Dysfunction It is also possible and contemplated to use the cyclic peptides of the present invention in combination with other drugs or agents, such as for the treatment of sexual dysfunction. These other drugs and agents may include agents that induce erectile activity, including phosphodiesterase-5 (PDE-5) inhibitors, testosterone, prostaglandins, etc. In a preferred embodiment of the present invention, the cyclic peptides of the present invention are used in combination with a therapeutically effective amount of a cyclic GMP-specific phosphodiesterase inhibitor or an alpha-adrenergic receptor antagonist. U.S. Patent No. 7,235,625, entitled "Multiple Agent Therapy for Sexual Dysfunction," discloses the use of cyclic peptides of the present invention in combination with other drugs or agents that induce erectile activity, such as phosphodiesterase-5 (PDE-5) inhibitors, testosterone, prostaglandins, etc. The teachings and disclosures are incorporated herein by reference as if fully set forth.

[0150] Thus, the present invention provides a method for treating sexual dysfunction, comprising administering a therapeutically effective amount of a cyclic peptide of the present invention in combination with a therapeutically effective amount of a second sexual dysfunction medication to a patient experiencing or at risk of experiencing sexual dysfunction. The cyclic peptide of the present invention can be administered simultaneously with, before, or after the administration of a therapeutically effective amount of the second sexual dysfunction medication. Preferably, the peptide of the present invention is administered within one hour, preferably within 30 minutes, of the administration of a therapeutically effective amount of the second sexual dysfunction medication. However, for certain forms of combination therapy, such as in combination with a therapeutically effective amount of a hormone or hormone-related dysfunction medication, the hormone or hormone-related dysfunction medication can be administered on an independent schedule, and there is no set or specific time relationship between the administration of the peptide of the present invention and the hormone or hormone-related dysfunction medication. Thus, for example, the hormone or hormone-related dysfunction medication can be administered daily or in other doses, or via a patch or other continuous administration schedule, along with the administration of the peptide of the present invention, as desired or required by the patient.

[0151] Thus, the present invention provides a method of treating sexual dysfunction, comprising administering to a patient having or at risk of having sexual dysfunction a therapeutically effective amount of a cyclic peptide of the present invention in combination with another compound useful in treating sexual dysfunction. In a preferred embodiment of the combination therapy, the sexual dysfunction is female sexual dysfunction, including but not limited to hypoactive sexual desire disorder. In another preferred embodiment of the combination therapy, the sexual dysfunction is erectile dysfunction.

[0152] The present invention also provides a pharmaceutical composition comprising a cyclic peptide of the present invention and a second compound useful for treating sexual dysfunction. In one embodiment of the present invention, the additional compound useful for treating sexual dysfunction is preferably selected from the group consisting of, but not limited to, phosphodiesterase inhibitors, cyclic GMP-specific phosphodiesterase inhibitors, prostaglandins, apomorphine, oxytocin modulators, α-adrenergic antagonists, androgens, selective androgen receptor modulators (SARMs), buproprion, vasoactive intestinal peptide (VIP), neutral endopeptidase inhibitors (NEP), and neuropeptide Y receptor antagonists (NPY).

[0153] In one embodiment of the present methods and compositions, the second sexual dysfunction medication is testosterone.

[0154] In another embodiment of the combination therapy, the second sexual dysfunction medication is a type V phosphodiesterase (PDE-5) inhibitor. For example, the PDE-5 inhibitor can be Viagra®, a trademark for sildenafil, Levitra®, a trademark for the monohydrochloride salt of vardenafil, or Cialis®, a trademark for tadalafil. Other PDE-5 inhibitors are described in U.S. Patent No. 7,235,625, entitled "Multiple Agent Therapy for Sexual Dysfunction," issued June 22, 2007, which is incorporated herein by reference. This will be disclosed in issue.

[0155] In another embodiment of the above composition, the compound useful for treating sexual dysfunction is an estrogen agonist / antagonist.In one embodiment, the estrogen agonist / antagonist is (-)-cis-6-phenyl-5-[-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-naphtho-talen-2-ol (also known as lasofoxifene) or its optical or geometric isomer; pharmaceutically acceptable salt, N-oxide, ester, quaternary ammonium salt; or prodrug thereof.More preferably, the estrogen agonist / antagonist is in the form of D-tartrate.

[0156] In another embodiment of the above composition, the estrogen agonist / antagonist is selected from the group consisting of tamoxifen, 4-hydroxytamoxifen, raloxifene, droloxifene, toremifene, centchroman, idoxifene, 6-(4-hydroxy-phenyl)-5-[4-(2-piperidin-1-yl-ethoxy)-benzyl]-naphthalen-2-ol, {4-[2-(2-aza-bicyclo[2.2.1]hept-2-yl)-ethoxy]-phenyl}-[6-hydroxy-2-(4-hydroxy-phenyl)-benzo[b]thiophen-3-yl]-methanone, EM-652, EM-800, GW5368, GW7604, TSE-424 and optical or geometric isomers thereof; and pharmaceutically acceptable salts, N-oxides, esters, quaternary ammonium salts and prodrugs thereof.

[0157] In yet another embodiment, the cyclic peptides of the present invention may be used in combination with any known mechanical aid or device.

[0158] 4.0 Methods of Administration and Use The methods of administration and use will depend on the characteristics of the particular peptides or formulas disclosed herein, the disease, indication, condition or syndrome to be treated and other factors known to those skilled in the art. The dosage and administration of the peptides disclosed herein or of the formulas disclosed herein may vary depending on other factors known in the art. Generally, any method of administration and use known in the art or developed thereafter may be used with the peptides disclosed herein or of the formulas disclosed herein. The following methods of administration and use have specific application to the indicated indications, without limitation to the foregoing.

[0159] 4.1 Subcutaneous use In one aspect, a composition comprising one or more peptides of the invention is formulated for subcutaneous injection, with the subcutaneous injections given at specified intervals, such as weekly or once or more times daily. In another aspect, the composition is formulated as an injectable sustained-release formulation. In one embodiment, the peptides of the invention are formulated with polyethylene glycol, e.g., polyethylene glycol 3350, and optionally one or more additional excipients and preservatives, including, but not limited to, excipients such as salts, polysorbate 80, sodium hydroxide or hydrochloric acid to adjust the pH. In another embodiment, the peptides of the invention are formulated with a poly(orthoester), which may be an autocatalytic poly(orthoester) with varying percentages of lactic acid in the polymer backbone, and optionally one or more additional excipients. In one embodiment, a poly(D,L-lactide-co-glycolide) polymer (PLGA polymer) is used, preferably a PLGA polymer with hydrophilic end groups, such as PLGA RG502H from Boehringer Ingelheim, Inc. (Ingelheim, Germany). For example, such a formulation can be made by combining the peptide of the present invention with a solution of PLGA in methylene chloride in a suitable solvent, such as methanol, under appropriate mixing conditions in a reactor, and then adding a continuous phase solution of polyvinyl alcohol thereto. Generally, any of a number of injectable biodegradable polymers can be used, which are preferably also adhesive polymers in sustained-release injectable formulations. The teachings of U.S. Patent Nos. 4,938,763, 6,432,438, and 6,673,767, and the biodegradable polymers and methods of formulation disclosed therein, are incorporated herein by reference. The formulations may require injections on a weekly, monthly, or other periodic basis, depending on the concentration and amount of peptide, the biodegradation rate of the polymer, and other factors known to those skilled in the art.

[0160] 4.2 Inhalation Use In one aspect, compositions comprising one or more peptides of the invention are formulated for administration to the respiratory tract (e.g., topically to the lungs and / or airways), alone or in combination with one or more inert carriers or additional active pharmaceutical ingredients, and in the form of a solution, suspension, aerosol, or dry powder formulation, such as an aerosol or solution for a nebulizer or as a fine powder for insufflation or inhalation. See generally Cryan, S.-A., "Carrier-based strategies for targeting protein and peptide drugs to the lungs," The AAPS Journal 7: E20-41 (2005). In general, the peptides of the invention may be used in the devices, formulations, compositions, and means described in one or more of the following U.S. patents or patent applications, each of which is incorporated herein by reference: U.S. Patent Application Publication No. 20090241949, "Dry powder inhalation system"; U.S. Patent Application Publication No. 20080066741, "Methods and systems of "Delivering medication via inhalation"; Patent No. 20070298116, "Amorphous, spray-dried powders having a reduced moisture content and a high long-term stability"; Patent No. 20070140976, "Aqueous inhalation pharmaceutical composition"; Patent No. 20060054166, "Inhalation nebulizer"; Patent No. 200 No. 50211244, "Dry powder preparations"; No. 20050123509. “Modulating charge density to produce improvements in the "Characteristics of spray-dried proteins"; U.S. Patent No. 20040241232, "Dry powder medicament formulations"; U.S. Patent No. 7,582,284, "Particulate materials"; U.S. Patent No. 7,481,212, "Increased dosage metered dose inhaler" ; US Patent No. 7,387,794, "Preparation of powder agglomerate"; US Patent No. 7,25 No. 8,873, "Preservation of bioactive materials by spray drying"; No. 7, No. 186,401, "Dry powder for inhalation"; No. 7,143,764, "Inhalation device"; No. 7,022,311, "Powdery inhalational preparations and process for producing the same"; No. 6,962,151, "Inhalation nebulizer"; No. 6,907,880, "Inhalation device"; No. 6,881,39 No. 8, “Therapeutic dry powder preparation”; No. 6,698,425, “Powder inhaler"; U.S. Pat. No. 6,655,380; "Inhalation device"; U.S. Pat. No. 6,645,4 No. 66, "Dry powder for inhalation"; No. 6,632,456, "Compositions for inhalation"; No. 6,610,272, "Medicinal aerosol formulation"; No. 6,596,261, "Method of administering a medicinal aerosol formulation"; No. 6,585,957, "Medicinal aerosol formulation"; No. 6,582,729, "Powered pharmaceutical formulations having improved dispersibility"; No. 6,572,893, "Systems and processes for spray drying hydrophobic drugs with hydrophilic excipients"; No. 6,551,578, "Modulated release particles for aerosol delivery"; No. 6,520,179, "Inhalation No. 6,518,239, "Dry powder compositions having improved dispersibility"; No. 6,503,481, "Compositions for aerosolization and inhalation"; No. 6,358,530, "Powdered pharmaceutical formulations having improved dispersibility"; No. 6,325,061, "Inhalation device" ; No. 6,257,232, "Inhalation device"; No. 6,187,344, "Powdered pharmaceutical formulations having improved dispersibility"; No. 6,116,237, "Methods of dry powder inhalation"; No. 5,934,272, "Device and method of creating aerosolized mist of respiratory drug"; and No. 5,558,085, "Intrapulmonary delivery of peptide drugs."

[0161] The composition may be a dry powder composition for localized delivery to the lungs via inhalation. The composition may contain a powder mixture for inhalation of the peptide of the present invention and a suitable powder base, diluent, or carrier substance, such as lactose, glucose, dextran, mannitol, or another sugar or starch. The composition may be used in any of a variety of dry powder devices, such as a reservoir dry powder inhaler, a multi-dose dry powder inhaler, or a nebulized metered-dose inhaler. The composition may contain additional excipients, such as alcohol, surfactants, lubricants, antioxidants, or stabilizers. Suitable propellants include hydrocarbon, chlorofluorocarbon, and hydrofluoroalkane propellants, or mixtures of any such propellants.

[0162] Inhalation solutions may also be formulated in liquid propellants for aerosol delivery, such as by pressurized metered dose inhalers. In yet another formulation, the solution may be in the form of an nebulized aqueous suspension or solution, with or without appropriate pH or isotonicity adjustment, as either a single dose or a multi-dose device.

[0163] 4.3 Nasal delivery Formulations or compositions suitable for nasal administration in which the carrier is a solid include coarse powders, e.g., with particle sizes in the range of 20 to 500 microns, administered in the manner in which snuff is administered (i.e., by rapid inhalation through the nasal passages from a container of powder held close to the nose). Suitable powder compositions include, by way of example, powdered preparations of the active ingredient thoroughly mixed with lactose or other inert powders acceptable for intrabronchial administration. Powder compositions may be administered via an aerosol dispersion device or may be contained in a breakable capsule that can be inserted by the patient into a device that punctures the capsule and expels the powder in a steady stream suitable for inhalation. Alternatively, suitable formulations may contain a liquid carrier, e.g., as a nasal spray or nasal drops, which may include an aqueous or oily solution of the active ingredient.

[0164] 4.4 Buccal and mucosal delivery Pharmaceutical compositions may further comprise, for example, one or more of water, a buffer (e.g., neutral buffered saline or phosphate buffered saline), ethanol, mineral oil, vegetable oil, dimethyl sulfoxide, a carbohydrate (e.g., glucose, mannose, sucrose, or dextran), mannitol, a protein, an adjuvant, a polypeptide or an amino acid such as glycine, an antioxidant, a chelating agent such as EDTA or glutathione, and / or a preservative. Additionally, one or more other active ingredients may (but need not) be included in the pharmaceutical compositions provided herein.

[0165] 4.5 Oral Delivery In one embodiment, peptides of the invention, including MC1r agonists, are orally administered and, in certain embodiments, delivered substantially intact to the lumen of all or a portion of the intestinal tract, including the patient's colon, for the treatment of inflammatory bowel disease, colitis, or other melanocortin receptor-mediated or -responsive gastrointestinal diseases, indications, conditions, and syndromes. Delayed-release polymer formulations comprising peptides of the invention, including, but not limited to, pH-dependent release polymers, may be used. The teachings and disclosures of WO 2019 / 023575 and WO 2019 / 183472, filed under the title "Melanocortin Receptor-Specific Formulations and Methods for Gastrointestinal Tract-Specific Delivery," are incorporated herein by reference as if fully set forth.

[0166] For systemic administration, compositions comprising one or more peptides disclosed herein or of formulas disclosed herein can be orally administered in individual dosage forms such as tablets or capsules. In a preferred embodiment, the individual dosage forms include an enteric coating and, optionally, one or more agents for increasing uptake, reducing protease degradation, improving cell permeability, etc. Any of a variety of delivery technologies can be used in oral delivery of the peptides of the present invention, including, but not limited to, liposomal compositions, mucoadhesive or gastroretentive delivery systems, absorption enhancers, multifunctional drug delivery systems, co-administration of penetration enhancers and / or protease inhibitors, covalent conjugation with various chemical or biological adjuvants, e.g., to improve cell permeability, enteric coatings, various nanoparticles, etc.

[0167] 5.0 Production Method In general, peptides disclosed herein or of the formulas disclosed herein can be synthesized by any means known in the art, including 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 can be used to prepare peptides disclosed herein or of the formulas disclosed herein.

[0168] Solid-phase peptide synthesis methods are well known and practiced in the art, and in such methods, synthesis of the peptides of the invention can be carried out by sequentially incorporating desired amino acid residues, one at a time, into a growing peptide chain according to the general principles of solid-phase methods.

[0169] In the chemical synthesis of peptides, the reactive side groups of the various amino acid residues are protected by appropriate protecting groups, which prevent chemical reactions from occurring at those sites until the protecting groups are removed. It is also common to protect the alpha-amino group of an amino acid residue or fragment while the moiety reacts with a carboxyl group, and then selectively remove the alpha-amino protecting group to allow subsequent reactions to occur at that site. Specific protecting groups are disclosed and known in the art for solid-phase and solution-phase synthesis.

[0170] Alpha amino groups can be protected by urethane-type protecting groups such as benzyloxycarbonyl (Z) and substituted benzyloxycarbonyl, e.g., p-chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p-biphenyl- It can be protected by suitable protecting groups including isopropoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc) and p-methoxybenzyloxycarbonyl (Moz) and aliphatic urethane-type protecting groups such as t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropoxycarbonyl and allyloxycarbonyl (Alloc), with Fmoc being particularly suitable for alpha amino protection.

[0171] The guanidino group can be protected with a suitable protecting group such as nitro, p-toluenesulfonyl (Tos), Z, pentamethylchromansulfonyl (Pmc), adamantyloxycarbonyl, pentamethyldihydrobenzofuran-5-sulfonyl (Pbf), and Boc. Pbf and Pmc are preferred protecting groups for Arg. Other reactive groups, including amine and carboxylic acid groups, can be similarly protected, such as 1-tert-butyl ester (OtBu) for Glu, Boc for Trp, and trityl (Trt) for His.

[0172] Linear peptide precursors to the peptides of the invention described herein were prepared using solid phase synthesis on an automated peptide synthesizer, using the programming module as provided by the manufacturer and following the protocols set forth in the manufacturer's manual.

[0173] Solid-phase synthesis begins at the C-terminus of the peptide. In one embodiment, a diamine linker or diamine ether linker is coupled to a suitable resin, thereby forming the starting resin, and then a protected alpha amino acid is coupled to the diamine or diamine ether linker. For example, such starting materials can be prepared by linking an alpha amino-protected amino acid to p-benzyloxybenzyl alcohol (Wang) resin, 2-chlorotrityl chloride resin, or oxime resin via a diamine or diamine ether linker. Diamine and diamine ether trityl resins are commercially available and are commonly used when commercially available.

[0174] Cyclic peptides disclosed herein or of the formulas disclosed herein can be readily synthesized by known conventional procedures for forming peptide linkages between amino acids. Such conventional procedures include, for example, any solution-phase procedure that allows condensation between the free alpha amino group of an amino acid residue having its carboxyl group and other reactive groups protected and the free primary carboxyl group of another amino acid residue having its amino group or other reactive groups protected. In a preferred conventional procedure, cyclic peptides disclosed herein or of the formulas disclosed herein can be synthesized by solid-phase synthesis and purified according to methods known in the art. Peptides disclosed herein or of the formulas disclosed herein can be prepared using any of a number of well-known procedures utilizing a variety of resins and reagents.

[0175] The process for synthesizing the present cyclic peptides can be carried out by a procedure in which each amino acid residue in the desired sequence is added sequentially, one at a time, to another amino acid residue, or by a procedure in which a peptide fragment having the desired amino acid sequence is first conventionally synthesized, and the resulting linear peptide is then cyclized in solution phase to yield the cyclic peptide of the invention.

[0176] In one embodiment, solid-phase synthesis is used to prepare the peptides of the invention. In this method, an N-terminal protecting group (PG), such as Fmoc or Boc, may be used, depending on the protecting group scheme used. Further orthogonal protection of the amino acid side chain bearing the reactive group may be used, e.g., Trp (Boc), Arg (Pbf), His (Trt), Glu (OtBu), etc. Generally, protecting groups are selected such that upon cleavage of the peptide from the solid-phase resin, the peptide is comprehensively deprotected, including removal of the orthogonal protecting group.

[0177] The solid support (shown in the diagram below by a bold circle) is provided with small molecules functionalized with the desired reactive group. In one embodiment, the reactive group comprises a diamine group, preferably selected from groups including: -NH-(CH2) 2~8 -NH-, or -NH-(CH2) 2~5 -O-(CH2) 2~5 -NH-.

[0178] In the scheme shown below, the reactive diamine group shown is -NH-(CH) 2~8 -NH- but the reactive group -NH-(CH2) 2~5 -O-(CH2) 2~5 It is understood that -NH- can be used as well.

[0179] Each amino acid to be coupled to the N-terminus of the peptide chain must be protected at its N-terminus and at its side chain if the amino acid contains a reactive group. The N-terminus can be protected by using any suitable protecting group; Boc and Fmoc are protecting groups commonly used in solid-phase peptide synthesis and can be used in the practice of the present invention as well. As can be seen below, solid-phase peptide synthesis traditionally proceeds by repeated cycles of alternating N-terminal deprotection and coupling reactions. However, unlike traditional solid-phase peptide synthesis using an amide resin, in one embodiment of the present invention, -NH-(CH) 2~8 -NH- or -NH-(CH2)2~5 -O-(CH2) 2~5 A resin with a diamine linker that may contain --NH-- is used.

[0180] In one embodiment, a pre-incorporated trityl group is used for ease of synthesis; for example, and not by way of limitation, the starting resin may be 1,4-diaminobutane trityl resin (ChemImpex, catalog number 04303), 1,2-Diaminoethane trityl resin (ChemImpex, catalog number 04306), or teeth 1,5-Diaminopentane trityl resin (ChemImpex, catalog number 04308) The trityl resin may further comprise a diamine linker group such as

[0181] However, other resins can be used, and other diamine or diamine ether linker groups can be used. Specific resins can be tailored to provide the desired diamine linker group, as needed. In one aspect, a resin such as 2-chloro-trityl is utilized, and the first step is to couple the selected diamine or diamine ether linker group to the resin before initiating synthesis. [ka]

[0182] As an example, the starting resin is 1,4-diaminobutane trityl resin (ChemImpex, catalog No. 04303, 0.77 mmol / g, 0.4 mmol) is loaded onto the peptide synthesizer. The next consecutive amino acid residue is the carboxy-terminal amino acid, which can be, for example, Trp. In that case, a protected Trp such as Trp(Boc) is used, resulting in -Trp(Boc)-NH-(CH)-NH-resin after deprotection of the -Trp(Boc) protecting group. It can be readily appreciated that other diamine linkers can be used, as can diamine ether linkers.

[0183] Each deprotection step may involve the use of, for example, piperidine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), 1-hydroxybenzotriazole (HOBT), N,N-dimethylformamide (DMF), or the like, followed by washing cycles with DMF or methyl tert-butyl ether (MBTE), or the like, optionally in repeated cycles.

[0184] Each coupling step was carried out using, among others, dichloromethane (DCM), HOBT, N,N-dichloromethane (N,N-dichloromethane), and HCl. This may involve using, for example, the desired protected amino acid, such as Fmoc-AA-OH, with coupling reagents including isopropylethylamine (DIPEA), DMF, or 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU). After coupling, a wash cycle may be used, such as with DMF or MBTE.

[0185] While the synthesized peptide is coupled to the resin or in solution, the N-terminus can be modified, for example, by acetylation. In one embodiment, after removal of the protecting group at the N-terminus, the resin-bound peptide is reacted with acetic anhydride in dichloromethane in the presence of an organic base such as diisopropylethylamine. Other methods of N-terminal acetylation are known in the art and can be used, including solution-phase acetylation.

[0186] The resulting resin-bound peptide can be cleaved from the resin by any means known in the art, such as by mixing the resin-bound peptide with a mixture of trifluoroacetic acid (TFA), triisopropylsilane (TIS), and water, e.g., a mixture of TFA / TIS / HO (95:2.5:2.5, v / v / v), at an appropriate temperature, e.g., room temperature, for an appropriate time, e.g., 20 minutes. If necessary, one or more additional cycles of mixing the resin-bound peptide with a mixture of TFA / TIS / HO can be performed after filtration. The combined filtrate can be stored, e.g., at room temperature, for 2 hours and then concentrated by purging with a stream of N. The cleaved linear peptide can then be precipitated from cold ether, and the resulting residue can then be dissolved in 50% t-butanol / water and lyophilized to yield the linear peptide.

[0187] The resulting crude linear peptide can then be cyclized in solution by conventional reaction procedures for cyclization through amide bond condensation. The linear peptide is first dissolved in a suitable solvent such as DMF, tetrahydrofuran (THF), DCM, or 1-methyl-2-pyrrolidone (NMP). Suitable cyclic coupling reagents include, for example, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), HBTU, benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate (BOP), benzotriazol-1-yl-oxy-tris(pyrrolidino)phosphonium hexafluorophosphate (PyBOP), 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TATU), 2-(2-oxo-1(2H)-pyridyl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TPTU), or N,N'-dicyclohexylcarbodiimide / 1-hydroxybenzotriazole (DCCI / HOBt). Coupling is conventionally initiated by the use of a suitable base such as DIPEA, sym-collidine or N-methylmorpholine (NMM).

[0188] After solution cyclization, the resulting mixture can be concentrated by known means and then partially purified, such as by trituration with methyl tert-butyl ether (MTBE). The resulting fractions can then be pooled and lyophilized.

[0189] For peptide No. 60 of the present invention, 1,4-diaminobutane trityl resin (ChemImpex, Cat. No. 04303, 0.77 mmol / g, 0.4 m The synthesis began with the loading of 100 mol of starting resin. The amino acids Fmoc-Trp(Boc), Fmoc-Arg(Pbf), Fmoc-D-Phe(4-F), Fmoc-His(Trt), Fmoc-Glu(OtBu), and Fmoc-Nle were coupled individually in sequence, with deprotection after each individual coupling. The Fmoc-Nle was deprotected, such as by using acetic anhydride and pyridine in DMF for an appropriate time, resulting in acylation of the amine to give the orthogonally protected, resin-bound linear peptide. Ac-Nle-Glu(OtBu)-His(Trt)-D-Phe(4-F)-Arg(Pbf)-Trp(Boc)-NH(CH2)4NH-resin

[0190] The peptide-resin was mixed with 8 mL of cleavage solution containing TFA / TIS / HO (95:2.5:2.5, v / v / v) for 20 min, which also cleaved the orthogonal protecting groups. The mixing was then repeated by mixing with 8 mL of fresh cleavage solution for another 20 min. The combined filtrate was stored at room temperature for 2 h and then concentrated to approximately 25 mL using a purged N2 stream. The cleaved linear peptide was precipitated from cold ether. The residue was dissolved in 50% t-butanol / water and lyophilized to yield the linear peptide (approximately 0.4 mmol). Ac-Nle-Glu-His-D-Phe(4-F)-Arg-Trp-NH(CH2)4NH2

[0191] The crude linear peptide was dissolved in a mixture of 3 mL of DMF and 3 mL of DCM, and the resulting solution was cooled in an ice-water bath. To the cold solution, 0.35 mL of EDC (0.18 g, 0.9 mmol) and HOAt (1.5 mL of a 0.6 M solution in DMF, 0.9 mmol) were added, followed by diisopropylethylamine (DIEA) (0.35 mL, 2.5 mmol) until the pH reached 9. The reaction mixture was stirred while warming to room temperature and continued stirring overnight at room temperature. LC / MS analysis indicated that the cyclization was complete. 2 mL of 1 N HCl was added to the reaction mixture, which was stirred for an additional 2 h, and then the pH was adjusted to pH 3–4. The pH-adjusted reaction mixture was directly loaded onto a preparative HPLC column, and the resulting pure fractions were pooled and lyophilized to afford 97.2 mg (19.8% yield) of the cyclic peptide. [ka]

[0192] C 18 Final purification may also be performed by reverse-phase high performance liquid chromatography (RP-HPLC) using an appropriate column such as a column, or other methods of separation or purification may be used, such as methods based on the size or charge of the peptide. Once purified, the peptides may be characterized by any of a number of methods, including high performance liquid chromatography (HPLC), amino acid analysis, mass spectrometry, etc.

[0193] Peptide No. 54 of the present invention was synthesized as shown above, substituting Met(O2) for His and D-Nal for D-Phe(4-F). After synthesis, cleavage from the resin and comprehensive deprotection were achieved by mixing with PyBop (3.3 g, 6 mmol) in 200 mL of a DMF / THF mixture (3:7, v / v), and the crude linear peptide was prepared. To initiate the cyclization (approximately 6 mmol), the resulting suspension was cooled in an ice-water bath. A mixture of PyBop (6.4 g, 12 mmol) and NMM (6.52 mL, 59.4 mmol, 9.9 eq) in 40 mL of a mixture of DMF / THF (3:7, v / v) was added to the cold solution. This was allowed to react for 3 h. The reaction mixture was concentrated on a rotary evaporator and then triturated with cold MTBE 3-4 times. The resulting oily residue was dissolved in 20 mL of 50% AcOH / water, stored overnight, and then subjected to HPLC purification. Pure fractions were pooled and lyophilized to give 670 mg of peptide No. 54 (9.7% yield).

[0194] Peptides Nos. 71 and 72 of the present invention were synthesized as described above for Peptide No. 60, except that each peptide consisted of five amino acids and the N-terminus was heptanoyl in the case of Peptide No. 71 and 3-phenylpropanoyl in the case of Peptide No. 72. In each case, after synthesis, cleavage from the resin, and global deprotection, cyclization of the crude linear peptide (approximately 3 mmol) was initiated by mixing with PyBop (1.6 g, 3 mmol) in 100 mL of a DMF / THF mixture (3:7, v / v). The resulting suspension was cooled in an ice-water bath. A mixture of PyBop (3.2 g, 6 mmol) and NMM (3.26 mL, 29.7 mmol, 9.9 eq) in 20 mL of a DMF / THF mixture (3:7, v / v) was added to the cold solution. The reaction mixture was allowed to react for 3 h, concentrated on a rotary evaporator, and then triturated with cold MTBE three or four times. The oily residue was dissolved in 10 mL of 50% AcOH / water, stored overnight, and then subjected to HPLC purification. Pure fractions were pooled and lyophilized to produce Peptide No. 71 (312 mg, 8.8% yield) and Peptide No. 72 (320 mg, 8.9% yield).

[0195] Peptide No. 77 was synthesized as described above for Peptide No. 60, except that Peptide No. 75 consisted of five amino acids and had a 2-naphthylacetyl N-terminus. Cyclization of the crude linear peptide (approximately 4 mmol) was initiated by mixing it with PyBop (4.16 g, 4 mmol) in 140 mL of a DMF / THF mixture (3:7, v / v), and the resulting suspension was cooled in an ice-water bath. A mixture of PyBop (4.16 g, 8 mmol) and NMM (6.52 mL, 39.6 mmol, 9.9 eq) in 20 mL of a DMF / THF mixture (3:7, v / v) was added to the cold solution. The reaction mixture was allowed to react for 3 h, concentrated on a rotary evaporator, and then triturated with cold MTBE three or four times. The oily residue was dissolved in 15 mL of 50% AcOH / water, stored overnight, and then subjected to HPLC purification. The pure fractions were pooled and lyophilized to yield Peptide No. 77 (586 mg, 11.8% yield).

[0196] Orthogonal protecting groups may also be used as needed. For example, peptides of the present invention contain multiple amino acids with amino-containing side chains. Any of a variety of protecting groups may be used, including allyl-Alloc protection schemes at specific amino acids and orthogonal protecting groups that are cleavable under different reaction conditions than are used for other amino acids with amino-containing side chains. Thus, for example, the Fmoc-Glu(OAll)-OH amino acid (Glu(OAll) refers to glutamic acid 5-allyl ester) may be used for the position that will be linked to the C-terminal diamine group upon cyclization, while other amino acids with amino-containing side chains may have different and orthogonal protecting groups, such as Fmoc-Arg(Pbf)-OH, Fmoc-Lys(Pbf)-OH, Fmoc-Dab(Pbf)-OH, etc. Other protecting groups can be used as well; by way of example and not limitation, with the side chain of His, Mtt (4-methyltrityl) or Mtt / OPp (4-methyltrityl / 2-phenylisopropyl) can be used, and for other positions that are not cleavable using conditions appropriate for cleavage of Mtt or Mtt / OPp, orthogonal protecting groups are utilized.

[0197] Reactive groups in the peptide can be selectively modified either during solid phase synthesis or after removal from the resin. For example, reactive groups in the peptide can be selectively modified while on the resin to obtain N-terminal modifications such as acetylation. The side chains may be modified or removed from the resin by use of a cleavage reagent and then modified. Similarly, methods for modifying the side chains of amino acids are well known to those skilled in peptide synthesis. The choice of modification made to reactive groups present on the peptide will be determined, in part, by the characteristics desired in the peptide.

[0198] While the synthesis is described primarily with reference to solid phase Fmoc chemistry, it is understood that other chemistries and synthetic methods can be used to make the cyclic peptides of the invention, including, by way of example and not limitation, any method that can be used utilizing solid phase peptide synthesis, Boc chemistry, solution chemistry, any of the various protecting group schemes, fragment condensation, any cyclization on or after removal from the resin, and other chemistries and synthetic methods.

[0199] 6.0 Formulations Depending on the desired route of administration, the formulation of the composition comprising one or more cyclic peptides disclosed herein or of the formula disclosed herein can vary.Thus, the formulation can be suitable for subcutaneous injection, sustained release subcutaneous injection, intravenous injection, nasal spray application, inhalation application, oral administration, including but not limited to oral release for the treatment of gastrointestinal diseases, buccal or other mucosal application, other transdermal application, etc.Generally, the formulation can be used for any form of administration of the peptide of the present invention.

[0200] 6.1 Salt Forms of Cyclic Peptides The cyclic peptides disclosed herein or of the formulas disclosed herein may be in the form of any pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutically acceptable non-toxic 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, etc. Particularly preferred are 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.

[0201] When a cyclic peptide disclosed herein or of a formula disclosed herein is basic, acid addition salts can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids, such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carboxylic acid, citric acid, ethanesulfonic acid, formic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, malonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, propionic acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, TFA, and the like. Acid addition salts of peptides disclosed herein or of formulas disclosed herein are prepared from the peptide and an excess of an acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, TFA, citric acid, tartaric acid, maleic acid, succinic acid, or methanesulfonic acid in a suitable solvent.

[0202] Acetate, ammonium acetate, and TFA salt forms are particularly useful. When a peptide disclosed herein or of a formula disclosed herein contains an acidic moiety, suitable pharmaceutically acceptable salts include alkali metal salts, such as sodium or potassium salts, or alkali metal salts. Examples of suitable polymorphs include alkali metal salts, such as calcium or magnesium salts. It should also be understood that certain peptides of Formulas I-V can exist in solvated and unsolvated forms, including solvates of the free peptide or solvates of a salt of the compound. The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term "hydrate" is used when the solvent is water. It should be understood that all polymorphs, including mixtures of different polymorphs, are included within the scope of the claimed peptides.

[0203] 6.2 Pharmaceutical Compositions The present invention provides a pharmaceutical composition comprising a cyclic peptide disclosed herein or of the formula disclosed herein and a pharmaceutically acceptable carrier.The carrier can be a liquid formulation, preferably a buffered, isotonic aqueous solution.Pharmaceutically acceptable carriers also include excipients, such as diluents, carriers, and additives, such as stabilizers, preservatives, solubilizers, buffers, etc., as described later in this specification.

[0204] The cyclic peptide compositions disclosed herein or of the formulas disclosed herein can be formulated or compounded into pharmaceutical compositions comprising at least one cyclic peptide disclosed herein or of the formulas disclosed herein, together with one or more pharmaceutically acceptable carriers, including excipients such as diluents, carriers, and additives such as stabilizers, preservatives, solubilizers, buffers, etc., if desired. Formulation excipients can include polyvinylpyrrolidone, gelatin, hydroxypropyl cellulose, acacia, polyethylene glycol, mannitol, sodium chloride, and sodium citrate. For injection or other liquid administration formulations, water containing at least one or more buffer components is preferred, and stabilizers, preservatives, and solubilizers can also be used. For solid administration formulations, any of a variety of thickeners, fillers, extenders, and carrier additives, such as starch, sugars, cellulose derivatives, fatty acids, etc., can be used. For topical administration formulations, any of a variety of creams, ointments, gels, lotions, etc. can be used. For most pharmaceutical formulations, inactive ingredients make up the greater portion of the preparation by weight or volume. It is also contemplated that any of a variety of metered release, sustained release, or extended release formulations and additives may be used for pharmaceutical formulations such that dosages may be formulated to provide delivery of the peptides of the formulas disclosed herein or disclosed herein over an extended period of time.

[0205] Generally, the actual amount of cyclic peptides disclosed herein or of the formulas disclosed herein administered to a patient will vary within a wide range depending on the mode of administration, the formulation used, and the desired response.

[0206] In practical use, the cyclic peptides disclosed herein or of the formulas disclosed herein can be combined as an active ingredient in admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral, parenteral (including intravenous), urethral, ​​vaginal, nasal, buccal, sublingual, etc. In preparing compositions for oral dosage form, any of the usual pharmaceutical media can be used, such as water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, etc. for oral liquid preparations such as suspensions, elixirs, and solutions; or carriers such as starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrating agents for oral solid preparations such as powders, hard and soft capsules, and tablets.

[0207] Tablets and capsules represent an advantageous oral dosage unit form due to their ease of administration. If desired, tablets can be coated by standard aqueous or non-aqueous techniques. The amount of active peptide in such therapeutically useful compositions is such that an effective dosage is obtained. In another advantageous dosage unit form, sublingual structures such as sheets, wafers, tablets, etc. can be used.

[0208] Tablets, pills, capsules, and the like may contain binders such as povidone, tragacanth, acacia, corn starch, or gelatin; diluents; fillers such as microcrystalline cellulose; excipients such as dicalcium phosphate; disintegrants such as corn starch, potato starch, or alginic acid; preservatives; coloring agents; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose, or saccharin. When the unit dosage form is a capsule, it may contain, in addition to the above-mentioned materials, a liquid carrier such as oil or fat. Various other materials may be used as coatings or to modify the physical form of the dosage form. For example, tablets may be coated with shellac, sugar, or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetener, methyl and propylparabens as preservatives, a pigment, and a flavoring such as cherry or orange flavor.

[0209] When formulated for oral delivery, the peptides can be formulated and prepared so that they are encapsulated in an enteric protective agent, more preferably so that they are not released until the tablet or capsule has passed into the stomach and, optionally, into the small intestine. In the context of this application, the term enteric coating or material is understood to refer to a coating or material that passes through the stomach essentially intact but disintegrates thereafter, releasing the active drug substance. Materials that can be used include cellulose acetate phthalate, hydroxypropylmethyl-ethylcellulose succinate, hydroxypropylmethylcellulose phthalate, polyvinyl acetate phthalate, and methacrylic acid-methyl methacrylate copolymer. The enteric coating used can be selected to promote dissolution of the dosage form primarily outside the stomach and to dissolve at a pH of at least about 5.5, more preferably from about 6.0 to about 8.0.

[0210] Any of a variety of permeation enhancers can be used to improve intestinal uptake upon dissolution of the enteric coating. In one aspect, the permeation enhancer enhances either the paracellular or transcellular transport system. Representative, non-limiting examples of such permeation enhancers include calcium chelators, bile salts (such as sodium cholate), and fatty acids. In some embodiments, a peptide or polypeptide that acts as a substrate for intestinal proteases is further added.

[0211] Cyclic peptides can also be administered parenterally. Solutions or suspensions of these active peptides can be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. These preparations can optionally contain preservatives to prevent the growth of microorganisms.

[0212] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.In all cases, the form must be sterile and fluid enough to be administered by syringe.The form must be stable under the conditions of manufacture and storage and must be protected from the contaminating action of microorganisms such as bacteria and fungi.The carrier can be, for example, a solvent or dispersion medium containing water, ethanol, polyols such as glycerol, propylene glycol or liquid polyethylene glycol, suitable mixtures thereof, and vegetable oils.

[0213] The cyclic peptides disclosed herein can be therapeutically administered via intranasal administration. The peptides can be in aqueous solutions, such as solutions containing saline, citrate, or other common excipients or preservatives, as well as absorption or penetration enhancers, transcellular penetration enhancers, mucoadhesive polymers, and various carrier systems. The peptides can also be in dry or powder formulations. The peptides disclosed herein can be in aqueous solutions, such as solutions containing saline, citrate, or other common excipients or preservatives, as well as absorption or penetration enhancers, transcellular penetration enhancers, mucoadhesive polymers, and various carrier systems. Cyclic peptides of the formulas described or disclosed herein can be formulated with any of a variety of agents that increase the effective nasal absorption of drugs, including peptide drugs. These agents can increase nasal absorption without unacceptable damage to the mucosa. U.S. Patent Nos. 5,693,608, 5,977,070, and 5,908,825, among others, teach many pharmaceutical compositions that may be used, including absorption enhancers; the teachings of each of the foregoing and all references and patents cited therein are incorporated by reference.

[0214] In aqueous solutions, the cyclic peptide may be appropriately buffered with saline, acetic acid, phosphate, citric acid, acetic acid, or other buffers, which may be at any physiologically acceptable pH, generally about pH 4 to about pH 7. Combinations of buffers, such as phosphate-buffered saline, saline, and acetate buffers, may also be used. In the case of saline, a 0.9% saline solution may be used. In the case of acetate, phosphate, citric acid, etc., a 50 mM solution may be used. In addition to buffers, appropriate preservatives may be used to prevent or limit bacterial and other microbial growth. One such preservative that may be used is 0.05% benzalkonium chloride.

[0215] In an alternative embodiment, the cyclic peptides disclosed herein or of the formulas disclosed herein can be administered directly to the lungs. Pulmonary administration can be achieved by a nebulized metered-dose inhaler, a device that, when activated by the patient during inspiration, allows for self-administration of a metered bolus of the peptides disclosed herein or of the formulas disclosed herein. In one aspect of this embodiment, the cyclic peptides can be in a dry, granular form, e.g., particles of about 0.5-6.0 μm, such that the particles have sufficient mass to settle on the lung surface and not be exhaled, but are small enough to prevent their deposition on airway surfaces before reaching the lungs. Any of a variety of different techniques can be used to create dry powder microparticles, including, but not limited to, micromilling, spray drying, and quick-freeze aerosolization followed by lyophilization. Using microparticles, the peptides can be deposited deep in the lungs, resulting in rapid and efficient absorption into the bloodstream. Furthermore, such an approach does not require penetration enhancers, as is sometimes the case with transdermal, nasal, or oral mucosal delivery routes. Any of a variety of inhalers can be used, including propellant-based aerosols, nebulizers, single-dose dry powder inhalers, and multi-dose dry powder inhalers. Common devices in current use include nebulized metered-dose inhalers, which are used to deliver medications for the treatment of asthma, chronic obstructive pulmonary disease, and the like. Preferred devices include dry powder inhalers, which are designed to form a cloud or aerosol of fine powder with particle sizes always less than about 6.0 μm.

[0216] Microparticle size, including mean particle size distribution, can be controlled by the method of preparation. In the case of micromilling, the size of the milling head, the speed of the rotor, the duration of the process, etc. control the microparticle size. In the case of spray drying, the nozzle size, flow rate, heat of the dryer, etc. control the microparticle size. In the case of preparation by quick-freeze aerosol followed by freeze-drying, the nozzle size, flow rate, concentration of the aerosolized solution, etc. control the microparticle size. These parameters and others can be used to control the microparticle size.

[0217] Cyclic peptides disclosed herein or of a formula disclosed herein may be therapeutically administered by injection of a sustained release formulation. In one embodiment, a cyclic peptide disclosed herein or of a formula disclosed herein is formulated for deep intramuscular injection, such as in the gluteal or deltoid muscle, with polyethylene glycol, such as polyethylene glycol 3350, and optionally one or more additional excipients and preservatives, including, but not limited to, excipients such as salts, polysorbate 80, sodium hydroxide or hydrochloric acid to adjust the pH. In another embodiment, a cyclic peptide disclosed herein or of a formula disclosed herein is formulated with a poly(orthoester), which may be an autocatalytic poly(orthoester) with any varying percentage of lactic acid in the polymer backbone, and optionally one or more additional excipients. In one embodiment, a poly(D,L-lactide-co-glycol) is formulated with a poly(D,L-lactide-co-glycol). In one embodiment, any of a number of injectable and biodegradable polymers, preferably also adhesive polymers, can be used in sustained-release injectable formulations. Alternatively, other sustained-release formulations can be used, including formulations that allow for subcutaneous injection, and other formulations can include one or more of nano / microspheres (such as compositions containing PLGA polymers), liposomes, emulsions (such as water-in-oil emulsions), gels, insoluble salts, or suspensions in oil. The formulations can be such that injections are required daily, weekly, monthly, or on other cycles, depending on the concentration and amount of cyclic peptide, the sustained-release rate of the substance used, and other factors known to those skilled in the art.

[0218] 6.3 Route of Administration When a composition comprising one or more peptides disclosed herein or of a formula disclosed herein is administered by injection, the injection can be intravenous, subcutaneous, intramuscular, intraperitoneal, or other means known in the art. The peptides disclosed herein or of a formula disclosed herein can be formulated by any means known in the art, including, but not limited to, as tablets, capsules, caplets, suspensions, powders, lyophilized preparations, suppositories, eye drops, skin patches, orally soluble formulations, sprays, aerosols, etc., and can be mixed with and formulated with buffers, binders, excipients, stabilizers, antioxidants, and other agents known in the art. Generally, any route of administration in which the peptides of the present invention are introduced across the epidermal layer of cells can be used. Thus, administration means can include administration via mucosal membranes, buccal administration, oral administration, dermal administration, inhalation administration, nasal administration, urethral administration, vaginal administration, etc.

[0219] 6.4 Therapeutically Effective Dose Generally, the actual amount of cyclic peptides disclosed herein or of the formula disclosed herein administered to a patient varies within a wide range, depending on the mode of administration, the formulation used, and the desired response. The dosage for treatment is administered by any of the above means or any other means known in the art in an amount sufficient to produce the desired therapeutic effect. The cyclic peptides disclosed herein or of the formula disclosed herein are generally highly active. For example, the cyclic peptide can be administered at about 0.001, 0.01, 0.1, 0.5, or 1 μg / kg body weight, depending on the specific peptide selected, the desired therapeutic response, the route of administration, the formulation, and other factors known to those skilled in the art.

[0220] 7.0 Tests and Assays Used in Evaluating Peptides The melanocortin receptor-specific peptides disclosed herein or of the formulas disclosed herein can be tested in a variety of assay systems and animal models to determine binding, functional status and efficacy.

[0221] 7.1 Assays for agonist activity performed at CEREP HTRF detection method was performed using CEREP (Eurofins CEREP SA, Celle-Levescault, France). The agonist activity of compounds at melanocortin receptors was determined by measuring their effect on cAMP production using a chromatographic assay. Cells were suspended in HBSS buffer (Invitrogen) supplemented with 20 mM HEPES (pH 7.4) and 500 μM IBMX, then distributed into microplates and incubated in the presence of HBSS (basal control), test compound, or reference agonist. Table 1 below includes incubation time, temperature, cell number, reference agonist, and cell line information. For stimulated control measurements, separate assay wells contained the reference compound. After incubation, cells were lysed, and a fluorescent acceptor (D2-labeled cAMP) and a fluorescent donor (anti-cAMP antibody labeled with europium cryptate) were added. After 60 minutes at room temperature, fluorescence transfer was measured using a microplate reader (Envision, Perkin Elmer) at an excitation wavelength of 337 nm and emission wavelengths of 620 and 665 nm. The (specific) cAMP concentration was determined by dividing the signal measured at 665 nm by that measured at 620 nm.

[0222] The results are expressed as a percentage of the control response to 1 μM of the reference. Several concentrations of the standard reference agonist were tested in each experiment to generate a concentration-response curve from which its EC 50 Calculate the value.

[0223] [Table 7]

[0224] 7.2 Assays for Antagonist Activity Performed at CEREP This assay was utilized to evaluate the antagonist activity of compounds at melanocortin receptors as determined by measuring their effect on cAMP production using HTRF detection methods.

[0225] The desired cells bearing melanocortin receptors were suspended in HBSS buffer (Invitrogen) supplemented with 20 mM HEPES (pH 7.4) and 500 μM IBMX and then distributed in a microplate in the presence of HBSS (basal control), test compound, or reference agonist. One concentration of agonist was then added to stimulate cAMP production. For basal control measurements, individual assay wells did not contain the reference agonist. Table 2 below includes incubation time, temperature, cell number, reference agonist, and cell line information.

[0226] After incubation, cells are lysed and a fluorescent acceptor (D2-labeled cAMP) and fluorescent donor (anti-cAMP antibody labeled with europium cryptate) are added. After 60 minutes at room temperature, fluorescence transfer is measured using a microplate reader (Envision, Perkin Elmer) at an excitation wavelength of 337 nm and emission wavelengths of 620 and 665 nm. The cAMP concentration is determined by dividing the signal measured at 665 nm by that measured at 620 nm (ratio). Results are expressed as percent inhibition of the control response to a reference agonist. A standard reference agonist is tested in each experiment at several concentrations to generate a concentration-response curve from which its IC 50 Calculate the value.

[0227] [Table 8]

[0228] 7.3 Alternative Assays for Agonist Activity Intracellular cAMP accumulation was examined as a measure of the peptide's ability to elicit a functional response in either HEK-293 cells expressing recombinant MC3r or MC4r or in B16-F10 (mouse) and HBL (human) cell lines expressing native MC1r. Confluent cells were detached from culture plates by incubation in enzyme-free cell dissociation buffer. Dispersed cells were suspended in Hank's balanced salt solution containing 10 mM HEPES (pH 7.5), 1 mM MgCl2, 1 mM glutamine, 0.5% albumin, and 0.3 mM 3-isobutyl-1-methyl-xanthine (IBMX), a phosphodiesterase inhibitor. 0.5 × 10 cells were cultured. 5 Cells were distributed into 96-well plates at a density of 10 cells / well and pre-incubated for 10 min. Cells were exposed to peptides dissolved in DMSO at concentrations ranging from 0.05 to 5000 nM (final DMSO concentration 1%) in a total assay volume of 200 μL for 15 min at 37°C. NDP-α-MSH was used as the reference agonist. Plates were read at 665 and 620 nM on a Perkin-Elmer Victor plate reader. cAMP levels were determined using the HTRF® cAMP cell-based assay system from Cisbio Bioassays, utilizing cryptate-labeled anti-cAMP and D2-labeled cAMP. Nonlinear analysis was performed using Graph-Pad Prism® software. Data analysis was performed by regression analysis. The maximum efficacy of the test peptide was compared to that achieved by the reference melanocortin agonist NDP-α-MSH.

[0229] 7.4 High and Low Density hMC4r Functional Assays The HEK293 cell line transfected with human MC4r (Palatin Technologies, US, licensed from the University of Michigan) was used. Human MC4r was introduced into HEK293 cells using the T-REx™ System (Invitrogen). The T-REx™ System transfects cells with tetracycline (Tet)-encoded E. coli Tn10. A tetracycline-regulated mammalian expression system is used, which uses regulatory elements from the resistance operon. The T-REx™ System allows expression of the gene of interest, the human MC4r gene. Expression of the gene is inhibited in the absence of tetracycline or doxycycline and is induced in the presence of tetracycline or doxycycline (see the T-REx™ System Manual published by Invitrogen).

[0230] L-glutamine (Gibco 25030) in 5% CO2 and 95% humidity at 37°C HEK293-T-REx-MC4r cells were cultured in DMEM (Gibco 11965) supplemented with 10% fetal bovine serum (FBS), 200 μg / mL Zeocin (Invitrogen 46-0072), and 6 mg / mL blasticidin (Invitrogen 46-1120). To induce hMC4r expression, T-150 flasks of cells at 75% confluence were incubated with two concentrations of doxycycline (0.1 ng / mL to provide a low-density hMC4r line and 10 ng / mL to provide a high-density hMC4r line) for 16-18 hours at 37°C in 5% CO2. On the day of the assay, cells were resuspended in PBS (Gibco 14190). Wash and harvest using cell dissociation buffer (Gibco 13150-016), followed by centrifugation. Separate and resuspend in Hank's Balanced Salt Solution (+Ca, +Mg) (Gibco 14025), 10 mM 4 -(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) (pH 7.4) (Sigma H0887), 1 mM L-glutamine (Gibco 25030), 1 mg / mL bovine serum albumin (BSA) (Sigma A3311), and 0.3 mM 3-isothiazolinone. The cells were resuspended in butyl-1-methyl-xanthine (IBMX).

[0231] Next, 198 μL of cells (approximately 5 × 10 4 ) / well into a 96-well plate (BD353916) and incubated for 10 minutes at 37°C. NDP-α-MSH was used as the reference agonist and was administered in a total assay volume of 200 μL. -5 ~10 -13 Cells were exposed to peptides dissolved in DMSO (final DMSO concentration 1%) at a range of concentrations for 15 min at 37°C. The reaction was stopped by adding 15 μL of lysis buffer / well, and the plates were shaken at room temperature for 30 min.

[0232] Plate reading on a Perkin-Elmer Victor plate reader at 665 and 620 nM cAMP levels were determined using the HTRF® cAMP cell-based assay system from Cisbio Bioassays, utilizing cryptate-labeled anti-cAMP and d2-labeled cAMP. Data analysis was performed by linear regression analysis. The maximum efficacy of the test peptide was compared to that achieved by the reference melanocortin agonist NDP-α-MSH.

[0233] Agonist stimulation of MC4r activates adenylate cyclase, an enzyme that catalyzes the formation of 3',5'-cyclic adenosine monophosphate (cAMP) from adenosine triphosphate (ATP). Therefore, agonist stimulation of MC4r increases cAMP levels. cAMP levels were measured using a cAMP dynamic 2HTRF kit (CisBio catalog number 62AM4PEC; see the manual published by CisBio). Plate controls (1% DMSO for 0% and 400 nM NDP-α-MSH for 100%) and a calibration curve ranging from 712 nM to 0.04 nM cAMP (as described in the CisBio HTRF kit) were included. cAMP levels were normalized to 0.25% CI 0.01 to 0.01 ... Next, calculate the fluorescence ratio as described in the CisBio HTRF kit and calculate the dose-response curve for the variable slope. Using the curve and based on the calculated cAMP concentration, the change in percent fluorescence was plotted against the cAMP concentration using GraphPad Prism software, and the EC 50 and E max value was determined.

[0234] 8.0 Example Peptide Structures In one aspect, cyclic peptides are provided that contain a core sequence derived from the sequence His-Phe-Arg-Trp within the cyclic portion, or modifications of that sequence, where the peptide is cyclized through the N-terminus, the side chain of the amino acid immediately adjacent to the His (or derivative, modification, or substitution of His) and the C-terminus of the peptide. The cyclic peptide is at least one cyclic pentapeptide containing five amino acids within the cyclic portion, and optionally a cyclic hexapeptide, hepta-peptide, or pentapeptide. It is a protapeptide or larger cyclic peptide, which has one or more additional amino acid residues outside the cyclic portion on the N-terminus.

[0235] In the case of MC4r antagonists, which may simultaneously contain MC1r, MC3r, or MC54 agonists, or combinations thereof, the core sequence derived from His-Phe-Arg-Trp in some embodiments may contain a Na1 or Na12 substitution at the Phe position, such as D-Phe at the Phe position rather than L-Phe, D-Nal1 or D-Nal2, or alternatively, a substituted Phe at the Phe position, such as a substituted L-Phe or D-Phe. A variety of amino acids may be utilized for the remaining amino acids in the core sequence. In general, the His position may be substituted or unsubstituted Pro, or an amino acid having a side chain containing at least one primary amine, secondary amine, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, alcohol, ether, sulfide, sulfone, sulfoxide, carbamoyl, or carboxyl. Arg positions can be substituted or unsubstituted Pro or can be an amino acid having a side chain containing at least one primary amine, secondary amine, guanidine, urea, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, or ether. Trp positions can be amino acids having a side chain containing at least one substituted or unsubstituted aryl or heteroaryl, or can alternatively be omitted.

[0236] Peptides encompassed within Formulas I, II, III, IV, and V contain one or more asymmetric elements, such as a chiral center or stereogenic axis, and peptides encompassed within such formulas may exist in different stereoisomeric forms. For both specifically described peptides, including those encompassed within Formulas I, II, III, IV, and V, and for generically described peptides, all forms of isomers at all chiral or other isomeric centers, including enantiomers and diastereomers, are intended to be encompassed herein. Each of the peptides of the present invention contains multiple chiral centers and may be used as a racemic mixture or an enantiomerically enriched mixture, in addition to using the peptides of the present invention in enantiomerically pure preparations. Generally, the peptides of the present invention are synthesized by the use of chirally pure reagents, e.g., the specified L- or D-isomer amino acids, using reagents, conditions, and methods to maintain enantiomeric purity; however, it is possible and contemplated that racemic mixtures may be produced. Such racemic mixtures may optionally be separated using well-known techniques, and the individual enantiomers may be used alone. Where a peptide can exist in tautomeric forms, and under particular conditions of temperature, solvent, and pH under which the peptide can exist in tautomeric forms, each tautomeric form, whether in equilibrium or predominantly in one form, is contemplated as being included within the present invention. Thus, single enantiomers of the peptides of Formulas I-V in optically active form can be obtained by asymmetric synthesis, synthesis from optically pure precursors, or resolution of the racemates.

[0237] Although the peptides disclosed herein are specific stereoisomeric forms of the peptides of Formulas I-V, the present invention should not be construed as being limited to the stereoisomeric forms encompassed by the peptides disclosed herein.

[0238] The present invention is further intended to include prodrugs of the peptides of the present invention, which undergo chemical conversion by metabolic processes before becoming active pharmacological peptides upon administration. Generally, such prodrugs are functional derivatives of the present peptides and are readily convertible in vivo to peptides of Formulas I-V. Prodrugs are any covalently bonded compounds that release the active parent peptide drug of Formulas I-V in vivo. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H. Bundgaard, Elsevier, 1985. Typical examples of prodrugs include those that contain a biologically labile protecting group on a functional site, for example, by esterification of a hydroxyl, carboxyl, or amino functional group. Thus, by way of example and not limitation, prodrugs include peptides of Formulas I-V in which an ester prodrug form is used, e.g., a lower alkyl ester of the R group of Formulas I-V, e.g., where R is -OH and the lower alkyl ester can have 1 to 8 carbons in the alkyl group or the aralkyl ester has 6 to 12 carbons in the aralkyl group. Broadly speaking, prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated in vivo to generate the active parent peptide drug of Formula I.

[0239] The present invention also includes peptides that are identical to those recited in Formulas I-V, except for the fact that one or more atoms set forth in such formulas are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes that may be incorporated into the peptides of the present invention include isotopes of hydrogen, carbon, nitrogen, and oxygen, e.g., 2 H, 3 H, 13 C. 14 C. 15 N, 18 O and 17O. Peptides disclosed herein or of the formulas disclosed herein and pharmaceutically acceptable salts or solvates of said peptides that contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of the present invention. Specific isotopically labeled peptides, e.g. 3 H and 14 Those incorporating radioactive isotopes such as C can be used in various assays, for example, in drug and / or substrate tissue distribution assays. 2 Substitution with heavier isotopes, such as with H, may provide pharmacological advantages in some instances, including improved metabolic stability. Isotopically labeled peptides of Formula I can generally be prepared by substituting an isotopically labeled reagent for a non-isotopically labeled reagent. [Example]

[0240] 9.0 Working Examples The present invention is further illustrated by the following non-limiting examples.

[0241] 9.1 Synthesized peptides and functional data 9.1.1 Synthesize a peptide of the following structure by the general method described above, and perform an EC 50 The values ​​were determined as shown. * EC marked with 50 Values ​​were determined by CEREP. "%" indicates EC 50 For values, the percent Emax (percent of the maximum response obtained with the positive control) is shown. EC for "NC" 50 The value is EC 50 Indicates values ​​greater than 10,000 nM and therefore not calculated.

[0242] [Table 9]

[0243] [Table 10]

[0244] Table 11

[0245] Table 12

[0246] Table 13

[0247] Table 14

[0248] Table 15

[0249] Table 16

[0250] Table 17

[0251] Table 18

[0252] Table 19

[0253] Table 20

[0254] Table 21

[0255] Table 22

[0256] Table 23

[0257] Table 24

[0258] Table 25

[0259] Table 26

[0260] Table 27

[0261] Table 28

[0262] Table 29

[0263] Table 30

[0264] Table 31

[0265] [Table 32]

[0266] [Table 33]

[0267] [Table 34]

[0268] [Table 35]

[0269] [Table 36]

[0270] [Table 37]

[0271] 9.1.1 The peptides of the following structures were synthesized by the general method described above, and the EC 50 The values ​​are determined as disclosed above. [ka] In the formula, Xaa 1 teeth, D-Phe(4-Cl), D-Phe(4-Me), D-Phe(4-OMe), D-Phe(4-Ph), D-Phe(4-NO2), D-Phe(4CN), D-Phe(3-Cl), D-Phe(3-Me), D-Phe(3-OMe), D-Phe(3-Phe), D-Phe(3-NO2), D-Phe(3-CN), D-Phe(2,4-diF), D-Phe(2,4-diMe), D-Phe(3,5-diF), D-Phe(3,4-diF), D-Phe(3,4-diOMe), D-Phe(2,4-diCl), D-Phe(2-Cl,4F), D-Nal1, D-Phe(2,3-diF), D-Phe(2,3-diCl), D-Phe(2,3-diMe), and D-Phe(2,3-diOMe) is.

[0272] Once synthesized, the peptides are tested as described herein and utilized in methods for preventing, ameliorating, or treating melanocortin receptor-mediated diseases, indications, conditions, and syndromes.

[0273] 9.2 Synthesis of Peptide No. 60 The peptide of Example 60 was synthesized for preclinical and clinical studies using standard solid-phase peptide chemistry to obtain a crude cyclic peptide intermediate. The resulting peptide intermediate was purified through a series of cleanup and preparative chromatography steps. Finally, the purified peptide was lyophilized prior to final packaging and storage. The following process overview flow diagram illustrates the synthetic method for the preparation of the acetate salt of the peptide of Example 60.

[0274] Steps 1~6 Fmoc-Trp(Boc)-NH-(CH2)4-NH-2Cl-Trt-Resin Complete cycles, with the following consecutive amino acids added in each cycle: Fmoc-Arg(Pbf)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-His(Trt)-OH, Fmoc-Nle-OH 1. Swelling: DMF 2. De-Fmoc: Piperidine / DBU / HOBt / DMF 3. Washing cycle: DMF, MBTE, DMF 4. Coupling cycle: DCM / Fmoc-AA-OH / HOBt, DIPEA, DMF, HBTU 5. Washing cycle: DMF, MBTE, DMF.

[0275] Step 7 Fmoc-Nle-Glu(OtBu)-His(Trt)-D-Phe(4-F)-Arg(Pbf)-Trp(Boc)-NH-(CH2)4-NH-2Cl-Trt-Resin Acetylation of resin-bound complete peptide sequences 1. Fmoc removal: piperidine / DBU / HOBt / DMF 2. Washing cycle: DMF, MBTE, DMF 3. N-terminal acetylation: Ac2-O / pyridine / DMF 4. Wash cycle: DMF / MBTE 5. Drying: Under vacuum, room temperature.

[0276] Step 8 Ac-Nle-Glu(OtBu)-His(Trt)-D-Phe(4-F)-Arg(Pbf)-Trp(Boc)-NH-(CH2)4-NH-2Cl-Trt-Resin Comprehensive cleavage and deprotection 1. Cutting: 95%TFA, 2.5%TIS, 2.5%H2O 2. Precipitation: MTBE (0-5°C) 3. Filtration and washing: MTBE 4. Drying: Vacuum drying at room temperature 5. Final washing and filtration: n-hexane 6. Drying: in vacuum at room temperature.

[0277] Step 9 Ac-Nle-Glu-His-D-Phe(4-F)-Arg-Trp-NH-(CH2)4-NH xTFA(x=1-3) Peptide Cyclization Dissolving: Linear peptides in THF / DMF and PyBOP Cyclization: PyBOP, NMM and THF / DMF Concentration: Rotary concentration Kenwa: EtOAc / MBTE Drying: Vacuum at room temperature.

[0278] Step 10 Ac-Nle-cyclo(Glu-His-D-Phe(4-F)-Arg-Trp-NH—(CH)—NH) x TFA (x=1-2) Ion exchange and product elution Dissolution and filtration: Cyclic peptide in TFA / water / ACN, filtration Column loading: Load onto a C18 silica gel column Elution: TFA / water / ACN gradient Salt exchange: 0.5M ammonium acetate Elution: AcOH, HO, ACN Lyophilization: Lyo flasks.

[0279] Final Ac-Nle-cyclo(Glu-His-D-Phe(4-F)-Arg-Trp-NH-(CH2)4-NH) xAcOH(x=1-2)

[0280] The peptide sequence was obtained using a condensation strategy in which protected amino acids were sequentially linked onto a solid support peptide resin. The amino acid coupling was performed using solid-phase peptide synthesis. where the next amino acid to be coupled to the N-terminus of the peptide chain is protected at its N-terminus and side chain using Fmoc protecting groups. The general SPPS procedure is one of repeated cycles of alternating N-terminal deprotection and coupling reactions, with resin washing between each step.

[0281] Coupling is monitored through the use of an in-process Kaiser test as well as a chloranil / toluene test, which produces a negative result, thereby indicating that the coupling reaction is complete. Once the reaction is complete, the reactor is drained and the resin is dried.

[0282] Once the complete peptide sequence was attached to the resin and the N-terminus acetylated, cleavage of the peptide resin was carried out in a glass reactor under temperature-controlled conditions using a cocktail of TFA, 3,6-dioxa-1,8-octanedioate (DODT), TIS, and water. After cleavage, the reaction mixture was filtered and washed with TFA. The peptide in the filtrate was precipitated using MTBE under temperature-controlled conditions and then filtered. The precipitated peptide was collected and triturated with n-hexane, and the crude linear peptide was dried under vacuum.

[0283] The resulting linear peptide was cyclized, followed by peptide precipitation and drying under vacuum. Cyclization was achieved by treating the linear peptide with the coupling agent benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate (PyBOP). The reaction was carried out at ambient temperature in the presence of a coupling medium, preferably a non-nucleophilic tertiary amine base such as N-methylmorpholine (NMM), in a mixture of tetrahydrofuran (THF) and dimethylformamide (DMF). The reaction was monitored by analytical RP HPLC testing. If the reaction was incomplete, additional aliquots of PyBOP and NMM were added to the reaction mixture, and the reaction was continued for complete cyclization. Upon completion of cyclization, the reaction mixture was concentrated on a rotary evaporator. The concentrate was triturated with a mixture of ethyl acetate and MTBE, stirred, and allowed to settle. The supernatant was decanted while the precipitate was collected. The solidified product was then isolated by filtration or centrifugation. The resulting solid cyclic peptide was dried under vacuum.

[0284] The crude cyclic peptide was purified using preparative reversed-phase high-performance chromatography (RP-HPLC) to obtain the final peptide product. Purification was achieved by co-elution of the peptide with a TFA, water, and acetonitrile (ACN) buffer system. Co-eluting fractions were analyzed for purity and impurity profile using an in-process control HPLC method. The purified peptide was converted to the acetate salt by salt exchange using acetic acid, acetonitrile, ammonium acetate, and water buffer solutions. The purified peptide was filtered and lyophilized using a manifold lyophilizer to remove water and remaining residual organic solvents.

[0285] The chemical formula of the resulting synthetic peptide is C as the acetate salt. 49 H 67 FN 14 O8 (CH3COOH (1≦x≦2), where x is a single peptide molecule). The exact mass of the resulting peptide, free of acetate, is 998.43 Da and the molecular weight is 999.16 Da.

[0286] 9.3 Formulation of Peptide No. 60 for Ophthalmic Use Peptide No. 60 is formulated as an ophthalmic solution drug product in a form-fill-dose container for topical administration for conditions such as dry eye disease. Manufacturing is aseptic, and sterile filtration utilizes redundant 0.2 μm sterilizing-grade polyethersulfone filters. The pH of the solution is approximately 6.5 for topical administration to the eye. The drug product is supplied in single-use form-fill ampoules, allowing for direct prodrug administration to the eye. Each ampoule contains a nominal volume of 0.25 mL. Secondary packaging is , foil pouches containing three ampoules in each pouch. The following table summarizes the formulation of the ophthalmic solution drug product.

[0287] [Table 38]

[0288] 9.4 Non-clinical studies of peptide number 60 for dry eye disease To investigate the effect of Peptide No. 60 on moderate chronic dry eye disease, male C57BL / 6JRj mice were treated with scopolamine (SiccaSystem™) and steroids. Animals were exposed to a controlled dry environment and disease was established before any treatment. Corneal fluorescein staining was performed on study days 10 and 12 (before treatment) and day 24 (12 days after topical treatment with the test compound). Based on the fluorescein score on day 12, animals were randomized into six different experimental groups (n=12 for each treatment group, n=7 for the untreated eye). Treatment arms included untreated and vehicle-treated animals and Peptide No. 60 (three different concentrations, 10 -2 , 10 -3 and 10 -4 Animals receiving either cyclosporine ophthalmic emulsion (mg / mL) or the reference compound sold under the trade name Restasis® were included.

[0289] Nonparametric Kruskal-Wallis ANOVA showed no statistical significance between experimental groups on day 12 (P = 0.84; Figure 1A). On day 24, corneal fluorescein scores were statistically significantly different between treatment groups (Kruskal-Wallis ANOVA, P < 0.001; Figure 1B). Restasis® (P < 0.001) and Peptide No. 60 (P < 0.001) were significantly different. -2 P<0.05 vs. mg / mL, 10 -3 and 10 -4 P < 0.01 vs. mg / mL) showed statistically significant lower corneal fluorescein staining compared with untreated eyes (Dunn's multiple comparisons test; Fig. 1B ).

[0290] Additionally, corneal fluorescein on day 12 was compared to day 24 for the same treated eyes. Based on this analysis, fluorescein staining in untreated eyes remained similar between days 12 and 24 (P=0.75). All experimental treatments (except vehicle) resulted in statistically significant improvements in corneal fluorescein staining (Figure 2).

[0291] 10 -2 ~10 -4 Peptide No. 60 at a dose of 100 mg / mL resulted in a statistically significant improvement in corneal fluorescein scores from days 12 to 24.

[0292] 9.5 Human Clinical Trials with Peptide No. 60 A multicenter, double-blind, randomized, placebo-controlled, parallel-group Phase 2 study enrolling approximately 160 subjects (80 subjects per treatment arm) utilized Peptide No. 60 in the ophthalmic solution formulation of Example 9.3. The purpose of the study was to evaluate the efficacy and safety of Peptide No. 60 in adult subjects with mild, moderate, or severe dry eye disease at Day 1 (baseline) in a controlled adverse environment (CAE) setting. The objective of this study was to compare the safety and efficacy of Peptide No. 60 to its vehicle from Day 1 (day 1) to Day 85 (week 12). Both signs and symptoms were assessed at regular clinic visits.

[0293] During the screening period, two 90-minute exposures to the CAE were administered to confirm eligibility for study participation. Qualified subjects were randomized to receive study drug or placebo in a double-blind manner for 12 weeks. Subjects self-administered eye drops three times daily (TID) and completed daily diary assessments as directed.

[0294] At Visits 3 (Day 15, Week 2), 4 (Day 29, Week 4), 5 (Day 57, Week 8), and 6 (Day 85, Week 12), subjects were exposed to CAE and ocular signs and symptoms were assessed pre-CAE, during CAE (symptoms only), and post-CAE.

[0295] In the overall enrollment population, which included patients with mild, moderate, or severe disease, statistical significance was not reached for the primary endpoint, improvement in inferior corneal staining (signs) and ocular discomfort (symptoms) as measured at the primary evaluation visit at week 12. In the moderate-to-severe patient population, statistically significant improvements in multiple signs and symptoms were achieved at the 2-week post-dose and 12-week visits. No safety signals were identified, and the Peptide No. 60 ophthalmic solution drug product was well tolerated in the eye.

[0296] In a subpopulation of moderate-to-severely affected patients (N=61), Peptide No. 60 ophthalmic solution drug product achieved statistical significance at weeks 2 and 12 (P<0.05 vs. vehicle) for multiple symptoms, including inferior (primary symptomatic endpoint), superior, and total corneal staining, temporal, nasal, and total conjunctival staining, and multiple ocular symptoms, including tear film breakup time and ocular discomfort. Additionally, multiple sign and symptom measures trended toward significance (P<0.1 vs. vehicle).

[0297] The study results demonstrated an excellent safety and tolerability profile. No serious adverse events related to study treatment were observed. Three patients in the placebo group and one patient in the Peptide No. 60 group (not considered drug-related) discontinued the study. There were no ocular drug-related adverse events in the Peptide No. 60 subjects.

[0298] 9.6 Comparison of Peptide No. 60 with α-MSH Potency of Peptide No. 60 (EC in nM) 50 The EC values ​​were compared to those of α-MSH. The profiles indicated greater relative potency of Peptide No. 60 at MC1r and MC5r compared to α-MSH, and the potency of Peptide No. 60 at MC3r and MC4r was broadly equivalent to that of α-MSH. Based on animal model studies, the efficacy of Peptide No. 60 offers a superior receptor profile for dry eye and other ocular diseases, while minimizing side effects due to MC3r and MC4r. Furthermore, virtually identical EC values ​​at MC1r and MC5r for Peptide No. 60 were observed. 50The EC value of α-MSH ensures simultaneous binding to both MC1r and MC5r. 50 From these values, it appears that other prior art melanocortin receptor peptides and compounds bind to MC1r but not to MC5r over a wide concentration range. Utilizing higher concentrations of α-MSH and other prior art melanocortin receptor peptides and compounds to bind to both MC1r and MC5r would result in undesired binding to MC3r and MC4r. In contrast, EC 50 The values ​​indicate that peptide no. 60 binds to MC1r and MC5r at selected concentrations without binding to MC3r and MC4r, resulting in a superior pharmacological response with minimal off-target effects.

[0299] [Table 39]

[0300] 9.7 Comparison of Peptide No. 60 with Bremeianotide The potency (EC in nM) of bremelanotide, a synthetic peptide of the sequence Ac-Nle-cyclo(Asp-His-D-Phe-Arg-Trp-Lys)-NH2 and peptide number 60 as disclosed in U.S. Pat. No. 6,579,968, which was recently approved for the treatment of hypoactive sexual desire disorder in premenopausal women. 50 values) were compared. This profile showed that peptide no. 60 was approximately equipotent at MC1r compared to bremeianotide, but bremeianotide lacked MC5r activity. The role of MC5r in dry eye is important both for the resolution of inflammation and also for tear structure, given that three glands that control secretion express MC5r - meibomian glands (lipid), lacrimal glands (aqueous), and conjunctival / goblet cells (mucus).

[0301] [Table 40]

[0302] 9.8 Stability and Pharmacokinetics of Peptide No. 60 Peptide No. 60 was evaluated in several stability and pharmacokinetic models. In a mouse model, Peptide No. 60 was found in both the conjunctiva and lacrimal gland at both 4 and 24 hours after ocular administration.

[0303] Although the present invention has been described in detail with particular reference to these preferred embodiments, other embodiments may achieve the same results. Variations and modifications of the present invention will be apparent 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 incorporated herein by reference.

Claims

1. Formula I: 【Chemistry 1】 (In the formula, Xaa １ is -R ５ -R ６ and Xaa ２ and Xaa ３ are each independently 1 to 3 L- or D-isomer amino acids or combinations thereof linked by backbone amide bonds, any backbone nitrogen atom optionally being C １ ~C ８ alkylated with a linear or branched alkyl, and when optionally alkylated, said alkyl optionally forms a ring with the side chain of the same amino acid when said side chain is aliphatic, said ring being optionally substituted; R １ is H or an L- or D-isomer amino acid side chain; R ２ is -(CH ２ ) ｕ - or - (CH ２ ) ｖ -O-(CH ２ ) ｗ - and; R ３ is H or C optionally containing one or more C═C double bonds １ ~C ９ a linear or branched aliphatic chain; R ４ is -H or C １ ~C ８ is a linear or branched alkyl; R ５ is optionally present, and when present, is one to three L- or D-isomer amino acids or combinations thereof linked by backbone amide bonds, any backbone nitrogen atom being optionally methylated; R ６ is H or C, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl or heteroaryl; １ ~C １７ is an acyl group; x is 1 to 5; u is 2 to 8; v is 2 to 5; and w is 2 to 5. or a pharmaceutically acceptable salt of any of the foregoing.

2. Xaa ２ -Xaa ３ 2. The peptide of formula I according to claim 1, wherein: comprises the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, wherein the phenyl ring of D-Phe or Phe is optionally substituted.

3. R １ 2. The peptide of formula I according to claim 1, wherein is the side chain of the L- or D-isomer amino acid Trp.

4. Xaa ２ -Xaa ３ comprises the amino acid sequence -His-D-Phe-Arg- or -His-Phe-Arg-, wherein the phenyl ring of D-Phe or Phe is optionally substituted, and R １ is the side chain of the L- or D-isomer amino acid Trp, peptides.

5. Xaa ２ -Xaa ３ 5. The peptide of claim 4, wherein the amino acid sequence -His-D-Phe-Arg- or -His-Phe-Arg- is optionally substituted on the phenyl ring of D-Phe or Phe.

6. Xaa ２ -Xaa ３ comprises the amino acid sequence -His-D-Phe- -His-Phe-, wherein the phenyl ring of D-Phe or Phe is optionally substituted, and R １ 2. The peptide of formula I according to claim 1, wherein is the side chain of the L- or D-isomer amino acid Arg.

7. R ５ 2. The peptide of formula I according to claim 1, wherein is the L- or D-isomer of Nle or Arg.

8. Xaa ２ -Xaa ３ 8. The peptide of claim 7, wherein the amino acid sequence -D-Phe-Arg- or -Phe-Arg- is optionally substituted in the phenyl ring of D-Phe or Phe.

9. Xaa ２ -Xaa ３ comprises the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, wherein the phenyl ring of D-Phe or Phe is optionally substituted, and R １ is the side chain of the L- or D-isomer amino acid Trp.

10. R ３ does not exist, and R ４ is C １ ~C １７ 2. A peptide of formula I according to claim 1, which is an acyl group.

11. Xaa ２ -Xaa ３ 11. The peptide of claim 10, wherein the phenyl ring of D-Phe or Phe is optionally substituted.

12. Xaa ２ -Xaa ３ comprises the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, wherein the phenyl ring of D-Phe or Phe is optionally substituted, and R １ is the side chain of the L- or D-isomer amino acid Trp.

13. R ５ is the L- or D-isomer of Nle or Arg, and R ６ is C １ ~C １７ 2. A peptide of formula I according to claim 1, which is an acyl group.

14. Xaa ２ and Xaa ３ 14. The peptide of claim 13, wherein the amino acid sequence -D-Phe-Arg- or -Phe-Arg- is optionally substituted in the phenyl ring of D-Phe or Phe.

15. Xaa ２ and Xaa ３ comprises the amino acid sequence -D-Phe-Arg- or -Phe-Arg-, wherein the phenyl ring of D-Phe or Phe is optionally substituted, and R １ is the side chain of the L- or D-isomer amino acid Trp.

16. The phenyl ring of D-Phe or Phe is optionally substituted with 1 to 3 ring substituents, which may be the same or different and each independently include halo, (C １ ~C １０ ) alkyl-halo, (C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, (C １ ~C １０ ) alkylthio, aryl, (C １ ~C １０ 15.) Any of claims 2, 4, 5, 6, 8, 9, 11, 12, 15 or 14, including alkylaryl, aryloxy, nitro, nitrile, sulfonamido, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy-carbonyl or aryloxy-carbonyl. The peptide according to any one of claims 1 to 4.

17. R ２ is -(CH ２ ) ｕ The peptide according to any one of claims 1 to 16, wherein

18. R ２ is -(CH ２ ) ｖ -O-(CH ２ ) ｗ The peptide according to any one of claims 1 to 16, wherein

19. 2. The peptide of claim 1, wherein the L- or D-isomer amino acids consist of alpha amino acids, beta amino acids, gamma amino acids, or delta amino acids, or a combination thereof.

20. Formula II: 【Chemistry 2】 (In the formula, Xaa １ is -R ５ -R ６ and R １ is substituted or unsubstituted indole, phenyl or naphthyl; R ２ is -(CH ２ ) ｕ - or - (CH ２ ) ｖ -O-(CH ２ ) ｗ - and; R ３ is H or C optionally containing one or more C═C double bonds １ ~C ９ a linear or branched aliphatic chain; R ４ is -H or -CH ３ and R ５ is optionally present, and when present, is 1 to 3 L- or D-isomer amino acids or combinations thereof, any backbone nitrogen atom optionally being methylated; R ６ is H or C, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl or heteroaryl; １ ~C １７ is an acyl group; R ７ is -H, -CH ３ or -CH ２ - and it is -CH ２ -, then R ８ Along with the general structure 【Transformation 3】 forming a ring of R ８ is R ８ is R ７ When forming the ring together with R, it is —H or ８ teeth, -(CH ２ ) ３ 、 -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -C(=O)-N(R １２ａ )(R １２ｂ )、 -O-(R １２ａ )、 -S-(=O) ２ -CH ３ 、 -S-(=O)-CH ３ 、 substituted or unsubstituted phenyl, -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; 【Chemistry 4】 and R ９ is a substituted or unsubstituted phenyl or naphthyl; R １０ teeth, -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -NH-C(=NH)-N(R １２ａ )(R １２ｂ )、 -NH-C(=O)-N(R １２ａ )(R １２ｂ )、 -O(R １２ａ )、 -C １ ~C １７ linear, branched or cyclic alkyl chains, -S(=O) ２ -CH ３ 、 -S(=O)-CH ３ 、 -C(=O)-O(R １２ａ )、 【Transformation 5】 and R １１ is -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; R １２ａ and R １２ｂ are each independently and in each instance independently H or C １ ~C ４ is a linear, branched or cyclic alkyl chain; t is independently in each instance 1 to 4; x is 1 to 5; u is 2 to 8; v and w are each independently 2 to 5; and z is 1 to 3.

2. The cyclic peptide of claim 1, wherein the cyclic peptide is:

21. R ９ is unsubstituted naphthyl.

22. Any substituted phenyl or naphthyl is, independently in each instance, substituted with 1 to 3 ring substituents, which may be the same or different and each independently include halo, (C １ ~C １０ ) alkyl-halo, (C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, (C １ ~C １０ ) alkylthio, aryl, (C １ ~C １０ 21. The cyclic peptide of claim 20, wherein said aryl is alkylaryl, aryloxy, nitro, nitrile, sulfonamido, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy-carbonyl, or aryloxy-carbonyl.

23. R ５ 21. The cyclic peptide of claim 20, wherein:

24. R ５ is a single L- or D-isomer amino acid having an aliphatic side chain.

25. The aliphatic side chain is —(CH ２ ) ３ -CH ３ 25. The cyclic peptide of claim 24, wherein:

26. R ５ is a single L- or D-isomer amino acid having a side chain containing at least one nitrogen atom.

27. R ５ is an L- or D-isomer of Arg, Lys, Orn, Dab, Dap, or Cit.

28. The cyclic peptide of formula II has the formula: 【Transformation 6】 21. The cyclic peptide of claim 20,

29. Xaa １ is Ac-Arg, Ac-D-Arg, Ac-Nle, Ac-D-Nle, Ac-Trp, cyclopentylacetyl, hexanoyl, 3-phenylpropanoyl, cyclopentylacetyl, acetyl, 2-naphthylacetyl, or 3-(1-naphthyl)propanoyl.

30. R １ is indole or naphthalene.

31. R ７ and R ８ The cyclic peptide of claim 20 or 28, wherein

32. 32. The cyclic peptide of claim 31, wherein the pyrrolidine is substituted.

33. The substituent is —O—CH ２ 33. The cyclic peptide of claim 32, wherein: -phenyl, wherein phenyl is substituted or unsubstituted.

34. R ９ is a substituted phenyl.

35. Formula III: 【Transformation 7】 (In the formula, Xaa １ is -R ５ -R ６ and R １ teeth, -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -NH-C(=NH)-N(R １２ａ )(R １２ｂ )、 -NH-C(=O)-N(R １２ａ )(R １２ｂ )、 -O(R １２ａ )、 -C １ ~C １７ linear, branched or cyclic alkyl chains, -S(=O) ２ -CH ３ 、 -S(=O)-CH ３ 、 -C(=O)-O(R １２ａ )、 【Transformation 8】 and R ２ is -(CH ２ ) ｕ - or - (CH ２ ) ｖ -O-(CH ２ ) ｗ - and; R ３ is H or C optionally containing one or more C═C double bonds １ ~C ９ a linear or branched aliphatic chain; R ４ is -H or -CH ３ and R ５ is optionally present, and when present, is 1 to 3 L- or D-isomer amino acids or combinations thereof, any backbone nitrogen atom optionally being methylated; R ６ is H or C, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl or heteroaryl; １ ~C １７ is an acyl group; R ７ is -H, -CH ３ or -CH ２ - and it is -CH ２ -, then R ８ Along with the general structure 【Chemistry 9】 Form a ring of: R ８ is R ８ is R ７ When forming the ring together with R, it is —H or ８ teeth, -H, -(CH ２ ) ３ 、 -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -C(=O)-N(R １２ａ )(R １２ｂ )、 -O-(R １２ａ )、 -S-(=O) ２ -R １２ａ 、 substituted or unsubstituted phenyl, -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; 【Chemistry 10】 and R ９ is a substituted or unsubstituted phenyl or naphthyl; R １１ is -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; R １２ａ and R １２ｂ are each independently and in each instance independently H or C １ ~C ４ is a linear, branched or cyclic alkyl chain; t is independently in each instance 1 to 4; x is 1 to 5; u is 2 to 8; v and 2 are each independently 2 to 5; and z is 1 to 3.

2. The peptide of claim 1, wherein the peptide is:

36. R ９ is unsubstituted naphthyl.

37. Any substituted phenyl or naphthyl is, independently in each instance, substituted with 1 to 3 ring substituents, which may be the same or different and each independently include halo, (C １ ~C １０ ) alkyl-halo, (C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, (C １ ~C １０ ) alkylthio, aryl, (C １ ~C １０ 36. The cyclic peptide of claim 35, wherein said aryl is alkylaryl, aryloxy, nitro, nitrile, sulfonamide, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy-carbonyl, or aryloxy-carbonyl.

38. R ５ 36. The cyclic peptide of claim 35, wherein:

39. R ５ is a single L- or D-isomer amino acid having an aliphatic side chain.

40. R ６ is acetyl, cyclopentylacetyl, hexanoyl, 3-phenylpropanoyl, cyclopentylacetyl, 2-naphthylacetyl, or 3-(1-naphthyl)propanoyl.

41. The cyclic peptide of formula III has the formula: 【Chemistry 11】 36. The cyclic peptide of claim 35,

42. Formula IV or V: 【Chemistry 12】 (In the formula, Z is H or an N-terminal group; Xaa １ is optionally present, and when present, is 1 to 3 amino acids, any backbone nitrogen atom of which is optionally methylated; Xaa ２ is the group -NH-(CH ２ ) ｕ -NH- or -NH-(CH ２ ) ｖ -O-(CH ２ ) ｗ an L- or D-isomer of an amino acid having a side chain containing a carboxyl group that forms an amide bond with the first —NH— of —NH—; Xaa ３ is an L- or D-isomer amino acid of Pro optionally substituted with hydroxyl, halogen, sulfonamide, alkyl, —O-alkyl, aryl, alkyl-aryl, alkyl-O-aryl, alkyl-O-alkyl-aryl, —O-alkyl-aryl or —O-aryl, or Xaa ３ is an L- or D-isomer amino acid having at least one primary amine, secondary amine, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, ether, sulfide, or carboxyl containing side chain; Xaa ４ is an L- or D-isomer amino acid with a substituted or unsubstituted aryl-containing side chain; Xaa ５ is an L- or D-isomer amino acid having a side chain containing at least one primary amine, secondary amine, guanidine, urea, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, or ether, and Xaa ６ is not present, the C-terminal carboxyl group is ２ ) ｕ -NH- or -NH-(CH ２ ) ｖ -O-(CH ２ ) ｗ -NH- forms an amide bond with the second -NH-; Xaa ６ is optionally present, and if present, is an L- or D-isomer amino acid having a side chain comprising at least one aryl or heteroaryl optionally substituted with one or more ring substituents, where one or more, if present, are the same or different and independently are hydroxyl, halogen, sulfonamide, alkyl, -O-alkyl, aryl, or -O-aryl, and the C-terminal carboxyl group is substituted with said group -NH-(CH ２ ) ｕ -NH- or -NH-(CH ２ ) ｖ -O-(CH ２ ) ｗ -NH- forms an amide bond with the second -NH-; u is 2 to 8; v and w are each independently 2 to 5; The group -NH-(CH ２ ) ｕ -NH- and -NH-(CH ２ ) ｖ -O-(CH ２ ) ｗ -NH- includes a first -NH- and a second -NH-. or a pharmaceutically acceptable salt thereof.

43. Z is a C alkyl group including linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl or aralkyl. １ ~C １７ 43. The cyclic peptide of formula IV or V of claim 42, wherein the N-terminal group is selected from the group consisting of acyl groups.

44. Xaa １ is Gly or L- or L- of Ala, Nle, Leu, Ile or Val 43. The cyclic peptide of formula IV or V of claim 42, wherein is a single amino acid residue selected from the group consisting of D-isomers.

45. Xaa １ 43. The cyclic peptide of formula IV or V of claim 42, wherein is a single amino acid having a side chain containing at least one primary amine, guanidine, or urea group.

46. Xaa １ 46. ​​The cyclic peptide of formula IV or V of claim 45, wherein is an L- or D-isomer of Arg, Lys, Orn, Dab, Dap, or Cit.

47. Xaa ４ 43. The cyclic peptide of formula IV or V of claim 42, wherein is D-Phe or Phe optionally substituted with 1 to 3 ring substituents.

48. The ring substituents may be the same or different and each independently may be selected from halo, (C １ ~C １０ ) alkyl-halo, (C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, (C １ ~C １０ ) alkylthio, aryl, (C １ ~C １０ 48. The cyclic peptide of formula IV or V of claim 47, wherein: 1) is alkylaryl, aryloxy, nitro, nitrile, sulfonamido, amino, monosubstituted amino, disubstituted amino, hydroxy, carboxy, or alkoxy-carbonyl.

49. Xaa ４ is D-Nal1 or D-Nal2.

50. Xaa ５ 43. The cyclic peptide of formula IV or V of claim 42, wherein is the L- or D-isomer of Arg, Lys, Orn, Dab, or Dap.

51. Xaa ６ is the L- or D-isomer of Trp, Na1 or Na12.

52. Z is C １ ~C ７ a linear alkyl acyl group; Xaa １ is the L- or D-isomer of Nle or Arg; Xaa ２ is the L- or D-isomer of Asp or Glu, and the carboxyl group of the side chain is the group -NH-(CH ２ ) ｕ -NH- or -NH-(CH ２ ) ｖ -O-(CH ２ ) ｗ -NH- forms an amide bond with the first -NH-; Xaa ３ is His, Hyp(Bzl), Met(O ２ ) or the L- or D-isomer of Asn; Xaa ４ is a substituted or unsubstituted Phe, L- or D-isomer of Na1 or Na12; Xaa ５ is the L- or D-isomer of Arg; and Xaa ６ is the L- or D-isomer of Trp, Na1 or Na12, the C-terminal carboxyl group of which is the group -NH-(CH ２ ) ｕ -NH- or -NH-(CH ２ ) ｖ -O-(CH ２ ) ｗ 43. The cyclic peptide of formula IV or V of claim 42, wherein the -NH- forms an amide bond with the second -NH-.

53. of formula IV, wherein Xaa ２ The carboxyl group of the side chain of ２ ) ｕ -NH of Xaa forms an amide bond with the first -NH-, and ６ The C-terminal carboxyl group of ２ ) ｕ 43. The cyclic peptide of claim 42, wherein the -NH- forms an amide bond with the second -NH-.

54. 43. The cyclic peptide of formula IV or V of claim 42, wherein at least one backbone nitrogen atom thereof comprises a methyl group.

55. 43. The cyclic peptide of formula IV or V of claim 42, wherein none of the backbone nitrogen atoms contains a methyl group.

56. structure: 【Chemistry 13】 or a pharmaceutically acceptable salt thereof.

57. 57. The cyclic peptide of claim 56, which is an acetate salt.

58. 57. The cyclic peptide of claim 56, which is a trifluoroacetate salt. 【Request Item 59】 【Chemistry 14】 or a pharmaceutically acceptable salt thereof.

60. 57. A pharmaceutical composition comprising the cyclic peptide of claim 56 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

61. 60. A pharmaceutical composition comprising the cyclic peptide of claim 59 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

62. 62. A method for the treatment of a melanocortin receptor-mediated disease, indication, condition or syndrome in a human or non-human mammal, comprising administering a pharmaceutical composition according to claim 60 or 61.

63. 62. A method for treating a condition responsive to altered melanocortin receptor function in a human or non-human mammal, comprising administering a pharmaceutical composition according to claim 60 or 61.

64. 64. The method of claim 63, wherein the melanocortin receptor-mediated disease, indication, condition or syndrome in a human is ocular inflammation.

65. 65. The method of claim 64, wherein the pharmaceutical composition is an aqueous pharmaceutical composition for administration to the ocular surface.

66. The inflammation is caused by an ocular condition, and the ocular condition may be dry eye disease, corneal ulcer, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scarring, epithelial defect, keratoconjunctivitis, idiopathic uveitis, corneal transplant, age-related macular degeneration, diabetic eye pathology, blepharitis, glaucoma, ocular hypertension, postoperative ocular pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascular membrane, vascular occlusive disease, allergic eye disease, tumor, retinal pigment epithelium ...

65. The method of claim 64, wherein the condition is a degeneration, an eye infection, scleritis, ptosis, miosis, eye pain, mydriasis, neuralgia, scarring eye surface disease, an eye infection, an inflammatory eye disease, an eye surface disease, a corneal disease, a retinal disease, an eye manifestation of a systemic disease, an inherited eye condition, an eye tumor, increased intraocular pressure, a herpetic infection, a pterygium, a persistent wound on the eye surface, eye pain and inflammation after laser photorefractive keratectomy, thermal or chemical burns to the cornea, a scleral wound, keratoconus, or a conjunctival wound.

67. 67. The method of claim 66, wherein the inflammation is caused by dry eye disease or keratoconjunctivitis sicca.

68. 62. The pharmaceutical composition of claim 60 or 61, wherein the pharmaceutically acceptable carrier is an aqueous solution comprising about 2.79 mg / mL trisodium citrate dihydrate, about 9 mg / mL sodium chloride, and about 1 mg / mL polysorbate 80.

69. 69. The pharmaceutical composition of claim 68, wherein the cyclic peptide is in trifluoroacetate salt form at a concentration of about 1.0 μg / mL.

70. 70. The pharmaceutical composition of claim 69, further comprising about 0.10 mg / mL anhydrous citric acid.

71. 69. The pharmaceutical composition of claim 68, wherein the aqueous solution has a pH of about 6.5, and the composition further comprises sodium hydroxide or hydrochloric acid as needed to adjust the pH.

72. 70. A method for treating dry eye disease or keratoconjunctivitis sicca, comprising administering the pharmaceutical composition of claim 69 at a frequency of about 3 times per day or less at a volume of about 50 μL or less per eye.

73. 73. The method of claim 72, wherein no more than about 150 ng of the cyclic peptide or a pharmaceutically acceptable salt thereof is administered per eye per day.

74. 1. A method for treating dry eye disease or keratoconjunctivitis sicca, comprising administering to the surface of each eye an EC50 concentration of less than 1 nM for each of MC1r and MC5r. ５０ 20. A method comprising administering an aqueous solution of a melanocortin receptor-specific peptide having a therapeutic value.

75. 75. The method of claim 74, wherein no more than about 150 ng of the melanocortin receptor-specific peptide is administered per eye per day as eye drops.

76. The melanocortin receptor-specific peptide is a peptide that binds to the EC of at least one of MC3r and MC4r in MC1r and MC5r. ５０ EC below value ５０ 75. The method of claim 74, The method described.

77. The melanocortin receptor-specific peptide has an EC2 activity that is at least four times lower in MC1r and MC5r than in at least one of MC3r and MC4r. ５０ 77. The method of claim 76, having a value.

78. The EC ５０ 78. The method of claim 77, wherein the values ​​are at least four times lower in MC1r and MC5r than in both MC3r and MC4r.

79. 75. The method of claim 74, wherein the peptide is either an agonist or a partial agonist at each of MC1r and MC5r.

80. EC ５０ 75. The method of claim 74, wherein the value is determined by cAMP production using an HTRF detection method such as that performed by CEREP.

81. 75. The method of claim 74, wherein the melanocortin receptor-specific peptide is a peptide described in claim 1.

82. The melanocortin receptor-specific peptide is 【Chemistry 15】 75. The method of claim 74, wherein:

83. structure 【Chemistry 16】 or a pharmaceutically acceptable salt thereof.

84. 84. The cyclic peptide of claim 83, which is an acetate salt.

85. 84. The cyclic peptide of claim 83, which is a trifluoroacetate salt. 【Request Item 86】 【Chemistry 17】 or a pharmaceutically acceptable salt thereof.

87. 84. A pharmaceutical composition comprising the cyclic peptide of claim 83 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

88. 87. A pharmaceutical composition comprising the cyclic peptide of claim 86 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

89. structure [Chemistry 18] or a pharmaceutically acceptable salt thereof.

90. 90. The cyclic peptide of claim 89, which is an acetate salt.

91. 90. The cyclic peptide of claim 89, which is a trifluoroacetate salt. 【Request Item 92】 【Chemistry 19】 or a pharmaceutically acceptable salt thereof.

93. 90. A pharmaceutical composition comprising the cyclic peptide of claim 89 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

94. 93. A pharmaceutical composition comprising the cyclic peptide of claim 92 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

95. Formula II: 【Chemistry 20】 (In the formula, Xaa １ is -R ５ -R ６ and R １ is substituted or unsubstituted indole, phenyl or naphthyl; R ２ is -(CH ２ ) ｕ - or - (CH ２ ) ｖ -O-(CH ２ ) ｗ - and; R ３ is H or C optionally containing one or more C═C double bonds １ ~C ９ a linear or branched aliphatic chain; R ４ is -H or -CH ３ and R ５ is optionally present, and when present, is 1 to 3 L- or D-isomer amino acids or combinations thereof, any backbone nitrogen atom optionally being methylated; R ６ is H or C, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl or heteroaryl; １ ~C １７ is an acyl group; R ７ is -H, -CH ３ or -CH ２ - and it is -CH ２ -, then R ８ Along with the general structure 【Chemistry 21】 forming a ring of R ８ is R ８ is R ７ When forming the ring together with R, it is —H or ８ teeth, -(CH ２ ) ３ 、 -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -C(=O)-N(R １２ａ )(R １２ｂ )、 -O-(R １２ａ )、 -S-(=O) ２ -CH ３ 、 -S-(=O)-CH ３ 、 substituted or unsubstituted phenyl, -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; 【Chemistry 22】 and R ９ is a substituted or unsubstituted phenyl or naphthyl; R １０ teeth, -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -NH-C(=NH)-N(R １２ａ )(R １２ｂ )、 -NH-C(=O)-N(R １２ａ )(R １２ｂ )、 -O(R １２ａ )、 -C １ ~C １７ linear, branched or cyclic alkyl chains, -S(=O) ２ -CH ３ 、 -S(=O)-CH ３ 、 -C(=O)-O(R １２ａ )、 【Chemistry 23】 and R １１ is -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; R １２ａ and R １２ｂ are each independently and in each instance independently H or C １ ~C ４ is a linear, branched or cyclic alkyl chain; t is independently in each instance 1 to 4; x is 1 to 5; u is 2 to 8; v and w are each independently 2 to 5; and z is 1 to 3. including all enantiomers, stereoisomers or diastereomers thereof or a pharmaceutically acceptable salt of any of the foregoing.

96. R ９ is unsubstituted naphthyl.

97. Any substituted phenyl or naphthyl is, independently in each instance, substituted with 1 to 3 ring substituents, which may be the same or different and each independently include halo, (C １ ~C １０ ) alkyl-halo, (C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, (C １ ~C １０ ) alkylthio, aryl, (C １ ~C １０ 96. The cyclic peptide of claim 95, wherein said aryl is alkylaryl, aryloxy, nitro, nitrile, sulfonamide, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy-carbonyl, or aryloxy-carbonyl.

98. R ５ 06. The cyclic peptide of claim 05, comprising at least one L- or D-isomer amino acid.

99. R ５ is a single L- or D-isomer amino acid having an aliphatic side chain.

100. The aliphatic side chain is —(CH ２ ) ３ -CH ３ 100. The cyclic peptide of claim 99, wherein:

101. R ５ is a single L- or D-isomer amino acid having a side chain containing at least one nitrogen atom.

102. R ５ is an L- or D-isomer of Arg, Lys, Orn, Dab, Dap, or Cit.

103. formula: 【Chemistry 24】 96. The cyclic peptide of claim 95,

104. Xaa １ is Ac-Arg, Ac-D-Arg, Ac-Nle, Ac-D-Nle, Ac-Trp, cyclopentylacetyl, hexanoyl, 3-phenylpropanoyl, cyclopentylacetyl, acetyl, 2-naphthylacetyl or 3-(1-naphthyl)propanoyl. The cyclic peptide of claim 95 or 103.

105. R １ is indole or naphthalene.

106. R ７ and R ８ The cyclic peptide of claim 95 or 103, wherein the cyclic peptide comprises pyrrolidine.

107. 107. The cyclic peptide of claim 106, wherein the pyrrolidine is substituted.

108. The substituent is —O—CH ２ -phenyl, wherein phenyl is substituted or unsubstituted.

109. R ９ is a substituted phenyl.

110. Formula III: 【Chemistry 25】 (In the formula, Xaa １ is -R ５ -R ６ and R １ teeth, -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -NH-C(=NH)-N(R １２ａ )(R １２ｂ )、 -NH-C(=O)-N(R １２ａ )(R １２ｂ )、 -O(R １２ａ )、 -C １ ~C １７ linear, branched or cyclic alkyl chains, -S(=O) ２ -CH ３ 、 -S(=O)-CH ３ 、 -C(=O)-O(R １２ａ )、 【Chemistry 26】 and R ２ is -(CH ２ ) ｕ - or - (CH ２ ) ｖ -O-(CH ２ ) ｗ - and; R ３ is H or C optionally containing one or more C═C double bonds １ ~C ９ a linear or branched aliphatic chain; R ４ is -H or -CH ３ and R ５ is optionally present, and when present, is 1 to 3 L- or D-isomer amino acids or combinations thereof, any backbone nitrogen atom optionally being methylated; R ６ is H or C, including optionally substituted linear or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl or heteroaryl; １ ~C １７ is an acyl group; R ７ is -H, -CH ３ or -CH ２ - and it is -CH ２ -, then R ８ Along with the general structure 【Chemistry 27】 forming a ring of R ８ is R ８ is R ７ When forming the ring together with R, it is —H or ８ teeth, -H, -(CH ２ ) ３ 、 -N(R １２ａ )(R １２ｂ )、 -NH-(CH ２ ) ｚ -N(R １２ａ )(R １２ｂ )、 -C(=O)-N(R １２ａ )(R １２ｂ )、 -O-(R １２ａ )、 -S-(=O) ２ -R １２ａ 、 substituted or unsubstituted phenyl, -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; 【Chemistry 28】 and R ９ is a substituted or unsubstituted phenyl or naphthyl; R １１ is -O-CH ２ -phenyl, where phenyl is substituted or unsubstituted; R １２ａ and R １２ｂ are each independently and in each instance independently H or C １ ~C ４ is a linear, branched or cyclic alkyl chain; t is independently in each instance 1 to 4; x is 1 to 5; u is 2 to 8; v and 2 are each independently 2 to 5; and z is 1 to 3. including all enantiomers, stereoisomers or diastereomers thereof or a pharmaceutically acceptable salt of any of the foregoing.

111. R ９ is unsubstituted naphthyl.

112. Any substituted phenyl or naphthyl is, independently in each instance, substituted with 1 to 3 ring substituents, which may be the same or different and each independently include halo, (C １ ~C １０ ) alkyl-halo, (C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, (C １ ~C １０ ) alkylthio, aryl, (C １ ~C １０ 111. The cyclic peptide of claim 110, wherein said aryl is alkylaryl, aryloxy, nitro, nitrile, sulfonamido, amino, monosubstituted amino, disubstituted amino, hydroxy, carbamoyl, carboxy, carbamoyl, alkoxy-carbonyl, or aryloxy-carbonyl.

113. R ５ 111. The cyclic peptide of claim 110, wherein said amino acid is at least one L- or D-isomer amino acid.

114. R ５ is a single L- or D-isomer amino acid having an aliphatic side chain.

115. R ６ is acetyl, cyclopentylacetyl, hexanoyl, 3-phenylpropanoyl, cyclopentylacetyl, 2-naphthylacetyl, or 3-(1-naphthyl)propanoyl.

116. formula: 【Chemistry 29】 111. The cyclic peptide of claim 110,

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