Reversibly amide-linked melanocortin receptor-specific cyclic peptides

Abstract

Disclosed herein are melanoreceptor-specific cyclic peptides having the formula: wherein Xaa 1 , R1, R2, R3, R4, R7, R8, R9, R 10 , t, x, and y are as defined in the specification; compositions and formulations comprising peptides having the above formula; and methods of preventing, ameliorating, or treating melanoreceptor-mediated diseases, indications, disorders, and syndromes using melanoreceptor-specific cyclic peptides having formula (I).

Description

[0001] Cross-references to related applications

[0002] This application claims priority and benefit to U.S. Provisional Patent Application Serial No. 62 / 969,315, filed February 3, 2020, entitled “Reverse Amide-Linked Melanocortin Receptor-Specific Cyclic Peptides,” the description and claims of which are incorporated herein by reference. Background Technology

[0003] Field of Invention (Technical Field):

[0004] This invention relates to melanocyte receptor-specific cyclic peptides (including cyclic peptides that are melanocyte receptor agonists, partial agonists, antagonists, or mixed agonist-antagonists) with reverse amide linkages from side chain to tail, and the use of melanocyte receptor-specific reverse amide linkage cyclic peptides in the treatment of melanocyte receptor-mediated diseases, indications, symptoms, and syndromes.

[0005] Relevant technical specifications:

[0006] The peptide has been cyclized via a two-amino acid side chain (one near the N-terminus of the peptide sequence and the other near the C-terminus), typically via a disulfide bond (e.g., via a side chain of two Cys residues) or an amide bond (e.g., via a side chain of two residues, one having a carboxyl group and the other having an amine group). Head-to-tail cyclized peptides are also known, such as peptides in which an amide-linked cyclic peptide is formed by coupling an N-terminal group (e.g., an amine) and a C-terminal group (e.g., a carboxylic acid).

[0007] A family of melanocyte-stimulating hormone (MSH) receptor types and subtypes has been identified, including MSH receptor 1 (MC1r), expressed on normal human melanocytes, melanoma cells, macrophages, and other cells; MSH receptor 2 (MC2r), expressing ACTH (adrenocorticotropic hormone), expressed on adrenal cells; MSH receptors 3 and 4 (MC3r and MC4r), expressed on cells in the hypothalamus, midbrain, brainstem, and peripheral tissues; and MSH receptor 5 (MC5r), expressed in widely distributed peripheral tissues. MC1r is believed to be involved in inflammation-mediated processes, hair and skin pigmentation, and other functions; MC2r is believed to mediate steroid production; MC3r is believed to be involved in energy homeostasis, feeding behavior, inflammation-mediated processes, and other functions; MC4r is believed to be involved in feeding behavior, energy homeostasis, sexual function, and other functions; and MC5r is believed to be involved in the regulation of the exocrine gland system and other functions.

[0008] Agonist and antagonist melanocortin receptor specific compounds are both known, including agonist and antagonist peptides. For example, MC4r agonist peptides are believed to be useful in the treatment of obesity or inducing weight loss, as well as in the treatment of various forms of sexual dysfunction, including male erectile dysfunction and female sexual dysfunction. MC4r antagonist peptides are believed to cause weight gain, and have potential utility in conditions such as cachexia and other wasting syndromes and conditions.

[0009] Peptide analogs of the endogenous agonist alpha-melanocyte stimulating hormone (a-MSH) are known. These peptide analogs include both linear peptides and cyclic peptides. Cyclic melanocortin receptor specific peptides are typically cyclized through a side chain (such as an amide or cysteine linkage), and are acylated at the N-terminus and amidated at the C-terminus (endogenous a-MSH is acylated at the N-terminus and amidated at the C-terminus), although a-MSH analogs are known that can have a C-terminal carboxyl group, as disclosed in U.S. Patent 6,579,968.

[0010] Despite the strong scientific and pharmaceutical interest in melanocortin receptor specific peptides (as evidenced by the numerous articles in the scientific literature and the numerous patent applications and issued patents), there are only three melanocortin receptor specific peptide drugs approved in the United States: bremelanotide (sold under the trade name for the indication of hypoactive sexual desire disorder in premenopausal women); amelanotide (sold under the trade name for the indication of photoprotection in adult patients with erythropoietic protoporphyria); and setmelanotide (sold under the trade name TM for the indication of the treatment of obesity due to proopiomelanocortin (POMC), proprotein convertase subtilisin / kexin type 1 (PCSK1), or leptin receptor (LEPR) deficiency). There remains a significant and substantial need for melanocortin receptor specific peptides for use in pharmaceutical applications, particularly in the treatment of inflammation-related diseases, indications, conditions, and syndromes. The present invention is made against this background. SUMMARY

[0011] In one aspect, the present invention relates to a cyclic peptide having the formula (I):

[0012]

[0013] including all enantiomers, stereoisomers, or diastereomers thereof, or a pharmaceutically acceptable salt of any of the foregoing,

[0014] wherein:

[0015] Xaa 1 is -R5-R6;

[0016] R1is a substituted or unsubstituted indole, phenyl, or naphthyl;

[0017] R2is -(CH2) u -;

[0018] R3is H or a C1to C9straight chain or branched aliphatic chain, the aliphatic chain optionally comprising one or more C=C double bonds;

[0019] R4is -H or -CH3;

[0020] R5is optionally present and, if present, is one to three L- or D-isomer amino acids, or combinations thereof, wherein any backbone nitrogen atom is optionally methylated;

[0021] R6is H or a C1to C 17 acyl group comprising an optionally substituted straight chain or branched alkyl, cycloalkyl, alkylcycloalkyl, aryl, aralkyl, or heteroaryl;

[0022] R7is -H, -CH3, or -CH2-, and if it is -CH2-, then forms a ring with R8having the general structure ;

[0023] If R8forms a ring with R7, then R8is -H, or R8is

[0024] -(CH2)3,

[0025] -N(R 12a )(R 12b ),

[0026] -NH-(CH2) z -N(R 12a )(R 12b ),

[0027] -C(=O)-N(R 12a )(R 12b ),

[0028] -O-(R 12a ),

[0029] -S-(=O)2-CH3,

[0030] -S-(=O)-CH3,

[0031] substituted or unsubstituted phenyl,

[0032] -O-CH2-phenyl (wherein the phenyl is substituted or unsubstituted),

[0033]

[0034] R9is substituted or unsubstituted phenyl or naphthyl;

[0035] R 10 is

[0036] -N(R 12a )(R 12b ),

[0037] -NH-(CH2) z -N(R 12a )(R 12b ),

[0038] -NH-C(=NH)-N(R 12a )(R 12b ),

[0039] -NH-C(=O)-N(R 12a )(R 12b ),

[0040] -O(R 12a ),

[0041] -C1to C 17 linear, branched, or cyclic alkyl chain,

[0042] -S(=O)2-CH3,

[0043] -S(=O)-CH3,

[0044] -C(=O)-O(R 12a ),

[0045]

[0046] R 11 is -O-CH2-phenyl (wherein the phenyl is substituted or unsubstituted);

[0047] R 12a and R 12b are each independently and in each instance independently H or a C1to C4linear, branched, or cyclic alkyl chain;

[0048] y is 0 or 1, and if 0, the group within the parentheses is not present, and if 1, the group within the parentheses is present;

[0049] t is independently in each instance 1 to 4;

[0050] x is 1 to 5;

[0051] u is 1 to 8; and

[0052] z is 1 to 3.

[0053] In one aspect, R9 is unsubstituted naphthyl. In another aspect, any substituted phenyl or naphthyl present in the cyclic peptide of Formula (I) is substituted in each instance independently with one to three ring substituents, where the substituents are the same or different and each is independently halo, (Ci-C 10 )alkyl-halo, (Ci-C 10 )alkyl, (Ci-C 10 )alkoxy, (Ci-C 10 )alkylthio, aryl, (Ci-C 10 )alkylaryl, aryloxy, nitro, nitrile, sulfonamide, amino, monosubstituted amino, disubstituted amino, hydroxyl, carbamoyl, carboxyl, carbamoyl, alkoxy-carbonyl, or aryloxy-carbonyl.

[0054] In one aspect of the cyclic peptide of Formula (I), R5 comprises at least one L- or D-isomer amino acid. In another aspect, R5 is a single L- or D-isomer amino acid having an aliphatic side chain, including where the aliphatic side chain is -(CH2)3-CH3. In another aspect, R5 is a single L- or D-isomer amino acid having a side chain comprising at least one nitrogen atom, including where R5 is an L- or D-isomer of Arg, Lys, Orn, Dab, Dap, or Cit.

[0055] The cyclic peptide of Formula (I) includes cyclic peptides having the following formula:

[0056]

[0057] In the cyclic peptide of Formula (I), R7 and R8 together can comprise a group:

[0058]

[0059] In the cyclic peptide of Formula (I), R8 can be -C(=0)-N(R 12a )(R 12b ), where R 12a and R 12b are H.

[0060] In the cyclic peptide of Formula (I), R8 can be an imidazole ring.

[0061] In the cyclic peptide of Formula (I), R5 can be absent, and in such cases, optionally R6 can be a C4 to C 17 acyl group.

[0062] In another aspect, the present application relates to a cyclic peptide having Formula (II):

[0063] In another aspect, the present application relates to a cyclic peptide having Formula (II):

[0064] or a pharmaceutically acceptable salt thereof, wherein

[0065] Z is H or an N-terminal group;

[0066] Xaa 1 is optionally present and, if present, is one to three amino acids, wherein any backbone nitrogen atom is optionally methylated;

[0067] Xaa 2 is an L- or D-isomer of an amino acid having a side chain comprising an amine group that forms an amide with the carboxyl group of Xaa 7 ;

[0068] Xaa 3 is an L- or D-isomer amino acid that is Pro, optionally substituted with a 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 a side chain comprising at least one primary amine, secondary amine, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, ether, sulfide, or carboxyl;

[0069] Xaa 4 is an L- or D-isomer amino acid having a side chain comprising a substituted or unsubstituted aryl group;

[0070] Xaa 5 is an L- or D-isomer amino acid having a side chain comprising at least one primary amine, secondary amine, guanidine, urea, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, or ether, and if Xaa 6 is not present, it has a C-terminal carboxyl group that forms an amide bond with the amine group of Xaa 7 ;

[0071] Xaa 6 is optionally present and, if present, it is an L- or D-isomer amino acid having a side chain comprising at least one aryl or heteroaryl group, optionally substituted with one or more ring substituents, and when one or more ring substituents are present, they are the same or different and independently a hydroxyl, halogen, sulfonamide, alkyl, -O-alkyl, aryl, or -O-aryl, and a C-terminal carboxyl group that forms an amide bond with the amine of Xaa 7 ; and

[0072] Xaa 7is an amino acid selected from the group consisting of glycine, beta-alanine, gamma-aminobutyric acid, 5-amino pentanoic acid, 6-amino hexanoic acid, 7-amino heptanoic acid, and 8-amino octanoic acid.

[0073] In the cyclic peptide of Formula (II), Z can be an N-terminal group selected from the group consisting of C1to C7linear alkyl, C3to C7branched alkyl, C3to C7cycloalkyl, C4to C7alkylcycloalkyl, C6to C10aryl, or C7to C12aralkyl. 17 acyl group.

[0074] In the cyclic peptide of Formula (II), Xaa 1 may be a single amino acid residue selected from the group consisting of Gly, or an L- or D-isomer of Ala, Nle, Leu, He, or Val. Alternatively, in the cyclic peptide of Formula (II), Xaa 1 may be a single amino acid having a side chain comprising at least one primary amine, guanidine, or urea group. Alternatively, in the cyclic peptide of Formula (II), wherein Xaa 1 may be an L- or D-isomer of Arg, Lys, Orn, Dab, Dap, or Cit.

[0075] In the cyclic peptide of Formula (II), Xaa 3 may be D-Phe or Phe, optionally substituted with one to three ring substituents. The ring substituents can be the same or different, and each is independently 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, hydroxyl, carboxyl, or alkoxy-carbonyl. Alternatively, in the cyclic peptide of Formula (II), Xaa 3 may be D-Nal 1 or D-Nal 2.

[0076] In the cyclic peptide of Formula (II), Xaa 5 may be an L- or D-isomer of Arg, Lys, Orn, Dab, or Dap.

[0077] In the cyclic peptide of Formula (II), Xaa 6 may be an L- or D-isomer of Trp, Nal 1, or Nal 2.

[0078] In one embodiment, in the cyclic peptide of Formula (II):

[0079] Z is a C1to C7linear alkyl acyl group;

[0080] Xaa 1 is an L- or D-isomer of Nle or Arg;

[0081] Xaa 2 is an L- or D-isomer of Dab, Dap, Orn, or Lys, wherein the side chain amine group forms an amide bond with the carboxyl of Xaa 7 ;

[0082] Xaa 3 is an L- or D-isomer of His, Hyp(Bzl), Met(O2), or Asn;

[0083] Xaa 4 is an L- or D-isomer of substituted or unsubstituted Phe, Nal 1, or Nal 2;

[0084] Xaa 5 is an L- or D-isomer of Arg; and

[0085] Xaa 6 is an L- or D-isomer of Trp, Nal 1, or Nal 2, wherein its C-terminal carboxyl group forms an amide bond with the amine of Xaa 7 .

[0086] The cyclic peptide of Formula (II) further includes the embodiments as described above, wherein at least one of its backbone nitrogen atoms comprises a methyl group.

[0087] In one aspect, a cyclic peptide template is provided that can be used to make a receptor-specific peptide of a biological receptor.

[0088] In another aspect, a melanocortin receptor-specific peptide-based pharmaceutical composition for use in treating melanocortin receptor-mediated diseases, indications, conditions, and syndromes is provided.

[0089] In another aspect, a melanocortin receptor-specific peptide-based pharmaceutical is provided, wherein the peptide is selective and is an agonist of MC1 r and an antagonist of MC4r.

[0090] In another aspect, a melanocortin receptor-specific peptide-based pharmaceutical is provided, wherein the peptide is selective and is an agonist of MC1 r and a partial agonist of MC4r.

[0091] In another aspect, a melanocortin receptor-specific peptide-based pharmaceutical is provided, wherein the peptide is selective and is an agonist of MC4r.

[0092] In another aspect, a receptor-specific peptide is provided that has a sub-nanomolar EC 50functional activity for one or more melanocortin receptors.

[0093] In another aspect, there is provided a melanocortin receptor specific peptide that is an agonist or partial agonist of one or more of MC1 r, MC3r and MC5r at an EC 50 value of less than 1 nM.

[0094] Additional aspects and novel features of the present application, as well as additional ranges of utility, will be set forth in part in the following detailed description, and will be in part apparent to those of ordinary skill in the art upon examination of the following detailed description, or can be learned by practice of the present application. These aspects of the present application can be realized and attained by means of the instrumentalities particularly pointed out in the appended claims and combinations thereof. DETAILED DESCRIPTION

[0095] 1.0 DEFINITIONS.

[0096] Before undertaking the description below, certain terms employed herein are defined as follows.

[0097] In the sequences given for the peptides disclosed herein, the amino acid residues have their conventional meanings as set forth in Manual of Patent Examining Procedure [Patent Examination Procedure Manual] the 9th edition of the Handbook of Biochemistry, Proteins, Volume 1 (1976) of the CRC Handbook of Biological Methods, Chapter 2400. 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 should be understood that D-isomers are designated by "D-" followed by the three letter code or amino acid name, such that, for example, D-Phe is D-phenylalanine. The foregoing unrecited amino acid residues include, but are not limited to, those having the following side chains, it being understood that such amino acid residues can be L-isomers or D-isomers:

[0098]

[0099]

[0100]

[0101]

[0102]

[0103] Amino acid residues further include, but are not limited to, the following, it being understood that such amino acid residues can be L-isomers or D-isomers:

[0104]

[0105]

[0106] The term "alpha amino acid" includes any amino acid having the general structure depicted in its non-ionized form, wherein R is any side chain group or hydrogen, including but not limited to the amino acid residues or side chain groups described in the above table and paragraph.

[0107] The term "L- or D-isomer amino acid" includes any isomeric form of any amino acid residue as defined herein, including in particular any alpha-, beta-, gamma-, or delta-amino acid, including but not limited to amino acids directly encoded by DNA, post-translationally modified amino acids, amino acids expressed by biological means other than directly from DNA, proprotein or non-proprotein amino acids, or any synthetic or artificial amino acid.

[0108] Amino acids, including L- or D-isomer amino acids, are linked together by "amide bonds" or amide linkages to form covalent peptide bonds that link the backbone carboxylic acid group of one amino acid to the backbone amino group of another amino acid, thereby forming a peptide bond (-C(=0)-NH-) or backbone amide bond.

[0109] The term "acyl" includes the group R(C=0)-, wherein R is an organic group, such as an alkyl, aryl, heteroaryl, carbocyclyl, or heterocyclyl group. When reference is made herein to a substituted acyl group, it is meant that the organic group (R) is substituted. Non-limiting examples of acyl groups include CH3-C(=0)-, referred to herein as an acetyl group or "Ac"; CH3-(CH2)4-C(=0)-, referred to herein as a hexanoyl group or "Hex"; CH3-(CH2)4-C(=0)-, referred to herein as a heptanoyl group or "Hept"; and various cyclic groups, such as phenylpropionoyl and cyclopentylacetyl.

[0110] A peptide or aliphatic moiety is "acylated" when an aliphatic group or substituted aliphatic group or aromatic group substituted aromatic group is bonded through a carbonyl {-(C=0)-} group to form an acyl group. Peptides are most commonly acylated at the N-terminus.

[0111] The term "alkane" includes straight chain or branched chain saturated hydrocarbons. Examples of straight chain alkane groups include methane, ethane, propane, and the like. Examples of branched or substituted alkane groups include methylbutane or dimethylbutane, methylpentane, dimethylpentane or trimethylpentane, and the like. In general, any alkyl group can be a substituent of an alkane.

[0112] The term "alkene" includes unsaturated hydrocarbons containing one or more double carbon-carbon bonds. Examples of such alkene groups include ethylene, propylene, and the like.

[0113] The term "alkenyl" includes straight chain monovalent hydrocarbon radicals of two to six carbon atoms containing at least one double bond or branched chain monovalent hydrocarbon radicals of three to six carbon atoms containing at least one double bond; examples include ethenyl, 2-propenyl, and the like.

[0114] The term "alkyl" as used herein includes those alkyl groups having the specified number of carbon atoms, which alkyl radicals are straight chain or branched chain saturated aliphatic hydrocarbon groups. C 1-10 Alkyl means an alkyl group having 1 to 10 carbon atoms. Non-limiting examples of such alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, t-butyl, pentyl, isopentyl, hexyl, isohexyl, and the like.

[0115] The term "alkyne" includes straight chain monovalent hydrocarbon radicals of two to six carbon atoms containing at least one triple bond or branched chain monovalent hydrocarbon radicals of three to six carbon atoms containing at least one triple bond; examples include ethyne, propyne, butyne, and the like.

[0116] The term "aryl" includes monocyclic or bicyclic aromatic hydrocarbon radicals having 6 to 12 ring atoms and optionally substituted independently with one or more substituents selected from alkyl, haloalkyl, cycloalkyl, alkoxy, alkylthio, 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.

[0117] The term "aralkyl" includes the group -R a R b where R a is an alkylene (divalent alkyl) group and R b is an aryl group as defined above. Examples of aralkyl groups include benzyl, phenethyl, 3-(3-chlorophenyl)-2-methylpentyl, and the like.

[0118] The term "aliphatic" includes compounds having hydrocarbon chains, such as, for example, alkyl, aryl, heteroaryl, alkane, alkene, alkyne, and derivatives thereof.

[0119] As used herein, the term "amide" includes compounds having a trivalent nitrogen attached to a carbonyl group, i.e., -C(=0)-NH2(i.e., a primary amide), -C(=0)-NHR c and -C(=0)-NR c R d where each of R c and R d independently represents hydrogen or an organic group. When reference is made herein to a substituted amide group, it is meant that at least one of the organic groups (R c and R d ) is substituted. Examples of amides include formamide, acetamide, propionamide, and the like.

[0120] An "amine" includes an amino group (-NH2), -NHR a and -NR a R b where each of R a and R b independently represents hydrogen or an organic group. When reference is made herein to a substituted amine group, it is meant that at least one of the organic groups (R a and R b ) is substituted.

[0121] A "nitrile" includes a compound that is a derivative of a carboxylic acid and contains a (-CN) group bound to an organic group.

[0122] The term "halogen" is intended to include the halogen atoms fluorine, chlorine, bromine, and iodine, as well as groups including one or more halogen atoms, such as -CF3, and the like.

[0123] The term "composition" as in pharmaceutical composition is intended to encompass a product comprising the active ingredient(s) and the carrier(s) constituting the bulk, as well as any product which results from the combination, complexation or aggregation of any two or more of the ingredients, or from the disintegration and / or dissolution or degradation of the ingredients. Thus, pharmaceutical compositions encompass any composition made by admixing an active ingredient with one or more pharmaceutically acceptable carriers.

[0124] By "agonist" with respect to a melanocortin receptor, it is meant an endogenous substance, a drug substance, or a compound (including certain peptide compounds disclosed herein) that can interact with a melanocortin receptor and elicit a pharmacological response, including but not limited to activation of the receptor, including initiation of signal transduction, such as adenylyl cyclase activation, melanocortin receptor characterization. A melanocortin receptor agonist can be an agonist of one or more of MC1 r, MC2r, MC3r, MC4r, and MC5r.

[0125] By "antagonist" with respect to a melanocortin receptor is meant an endogenous substance, a drug substance, or a compound (including certain peptide compounds disclosed herein) that blocks or inhibits the action of an agonist at a melanocortin receptor. A melanocortin receptor antagonist can be an antagonist of one or more of MC1 r, MC2r, MC3r, MC4r, and MC5r. Certain compounds (including certain peptide compounds disclosed herein) can be an agonist of one or more melanocortin receptors and an antagonist of one or more other melanocortin receptors.

[0126] By "a-MSH" is meant the peptide Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH2 and analogs and homologs thereof, including but not limited to NDP-a-MSH.

[0127] By "NDP-a-MSH" is meant the peptide Ac-Ser-Tyr-Ser-Nle-Glu-His-D-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH2 and analogs and homologs thereof.

[0128] By "EC 50 " is meant the molar concentration of an agonist (including partial agonist) that produces 50% of the maximum possible response for that agonist. For example, if a test compound at a concentration of 72 nM produces 50% of the maximum possible response for that compound, as determined in a cAMP assay in an MC4r cell expression system, then the EC 50 of that compound is 72 nM. Unless otherwise indicated, molar concentrations associated with an EC 50 determination are in nanomoles per liter (nM).

[0129] By "Ki (nM)" is meant the equilibrium inhibitor dissociation constant, which indicates the molar concentration of a competing compound that binds to one-half of the binding sites of a receptor at equilibrium in the absence of competitors. In general, the value of Ki is inversely related to the affinity of a compound for a receptor, so if the Ki is low, the affinity is high. Ki can be determined using the equation of Cheng and Prusoff (Cheng Y., Prusoff W.H., Biochem. Pharmacol. 22:3099-3108, 1973):

[0130]

[0131] Unless otherwise indicated, molar concentrations associated with a Ki determination are in nM. Ki can be expressed with respect to a particular receptor (e.g., MC1 r, MC3r, MC4r, or MC5r).

[0132] "Inhibition" means a decrease or percentage decrease in receptor binding in a competitive inhibition assay as compared to a known standard. Thus, "inhibition at 1 μM (NDP-α-MSH)" means the percentage decrease in NDP-α-MSH binding by the addition of a defined amount of test compound (e.g., 1 μM of test compound) under the assay conditions described below. For example, a test compound that does not inhibit NDP-α-MSH binding has 0% inhibition, while a test compound that completely inhibits NDP-α-MSH binding has 100% inhibition. Typically, a detectably labeled assay is used for competitive inhibition testing (e.g., with I 125 labeled NDP-α-MSH) or lanthanide chelate fluorescence assays (e.g., with Eu-NDP-α-MSH). However, other methods of testing competitive inhibition are known, including the use of different labeling or tagging systems, and generally any method known in the art for testing competitive inhibition can be used in the present application. It will thus be appreciated that "inhibition" is a measure of whether a test compound attenuates the binding of α-MSH to the melanocortin receptor.

[0133] "Binding affinity" means the ability of a compound or drug to bind to its biological target, herein expressed as Ki (nM).

[0134] "E max " means the maximum functional activity that a compound can achieve in a particular melanocortin receptor expressing cell system, such as the maximum stimulation of adenylyl cyclase. The maximum stimulation achieved by NDP-α-MSH is assigned as 100% E max , while a compound that is able to stimulate half of the maximum activity of NDP-α-MSH is assigned as having 50% E max . E max max is 70% or greater can be classified as an agonist, E max a compound between 10% and 70% can be classified as a partial agonist, while E max a compound less than 10% can be classified as inactive.

[0135] In general, "functional activity" is a measure of receptor signaling, or a change in signaling associated with a receptor (e.g., the melanocortin receptor), after the receptor has been activated by a compound. The melanocortin receptor initiates signal transduction by activating a heterotrimeric G protein. In one aspect, the melanocortin receptor signals through Gα S , which in turn activates adenylyl cyclase. In another aspect, the melanocortin receptor signals through Gα ScAMP is generated through catalysis by adenylyl cyclase. Thus, determining stimulation of adenylyl cyclase (e.g., determining maximal stimulation of adenylyl cyclase) is one measure of functional activity, and is the primary measure exemplified herein. However, it will be appreciated that alternative measures of functional activity can be used in the practice of the present application, and are specifically contemplated and included within the scope of the present application. Thus, in one example, intracellular free calcium can be measured using a specific fluorescent molecule that binds to calcium (e.g., Fura2) and methods disclosed in Mountjoy K.G. et al., Melanocortin receptor-mediated 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 a commonly used alternative calcium-binding dye (Nohr et al., The orphan G protein-coupled receptor GPR139 is activated by the peptides: Adrenocorticotropic hormone (ACTH), alpha-, and beta-melanocyte stimulating hormone (alpha-MSH, and beta-MSH), and the conserved core motif HFRW. Neurochem Int 102: 105-113, 2017). In Ca2 +Upstream of the release event and in the same pathway, activation can also be measured by measuring inositol triphosphate or diacylglycerol produced from phosphatidylinositol 4,5-bisphosphate, as in a commercially available HTRF assay (Liu et al., Comparison on functional assays for Gq-coupled GPCRs by measuring inositol monophospate-1 and intracellular calcium in 1536-well plate format. Curr Chem Genomics 1 :70-77, 2008). Yet another measure of functional activity is receptor internalization resulting from activation of the regulatory pathway, as with the methods disclosed in Nickolls S.A. 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. Yet another measure of functional activity is the exchange and rate of exchange of nucleotides associated with G protein receptor activation, such as the exchange of GDP (guanosine diphosphate) for GTP (guanosine triphosphate) on the G protein alpha subunit, which can be measured by any means, including using guanosine 5'-(gamma- 35S]Radioassay of (thio)-triphosphates as disclosed in Manning D.R., Measures of efficacy using G proteins as endpoints: differential engagement of G proteins through single receptors. Mol Pharmacol 62: 451-452, 2002. A relatively new assay platform was designed to measure the activity / binding of 14 different Ga species belonging to the Gi, Gq, Gs, Gi2 / i3 subfamilies as they relate to receptors, which uses a BRET (Bioluminescence Resonance Energy Transfer)-based biosensor to measure the dissociation of Ga and Gy 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 to measure the activation of G-coupled proteins, such as those disclosed below: 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 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 orphanseven-transmembrane. [Improved receptor gene assay for identifying ligands acting on orphan seven-transmembrane receptors]. Pharmacology & Toxicology 93:249-258, 2003. The colorimetric assay by Chen et al. has been adapted for use in measuring melanocyte-stimulating receptor activation, as disclosed in: Hruby VJ et al., Cyclic lactamα-melanocortin analogues of Ac-Nle. 4 -cyclo[Asp 5 ,D-Phe 7 Lys 10] α-melanocyte-stimulating hormone-(4-10)-NH2 with bulky aromatic amino acids at position 7 shows high antagonist potency and selectivity at specific melanocortin receptors. 4 - Ring [Asp] 5 ,D-Phe 7 Lys 10]Cyclic lactam alpha-melanocyte-stimulating hormone analogs of alpha-melanocyte-stimulating hormone-(4-10)-NH2 exhibit high antagonist potency and selectivity for specific melanocortin receptors. J Med Chem 38:3454-3461, 1995. Generally, functional activity can be measured by any method, including methods of determining activation of G-coupled receptors and / or signaling, and further including methods that can be developed or reported later. Each of the foregoing clauses and methods disclosed therein are incorporated by reference herein as if fully set forth.

[0136] As used herein, the terms "treat," "treating," and "treatment" refer to actions that occur when a patient is suffering from a particular disease or disorder, which reduce the severity of that disease or disorder.

[0137] As used herein, the term "pharmacologically effective amount" (including "therapeutically effective amount") means an amount of a peptide according to the present application sufficient to elicit a desired therapeutic effect or biological effect.

[0138] As used herein, the term "therapeutically effective amount" means an amount of a compound, including a peptide of the present application, which will elicit a biological or medical response in a mammal treated by a medical practitioner or other clinician.

[0139] As used herein, the term "prophylactically effective" or "prophylactically" means an amount of a compound, including a peptide of the present application, which can prevent or inhibit, or lessen the suffering of a mammal having a medical condition that a medical practitioner or other clinician is attempting to prevent, inhibit, or alleviate in a patient prior to the onset of the particular disease or disorder.

[0140] 2.0 Clinical Indications and Utility.

[0141] The compositions and methods disclosed herein can be used for medical applications as well as for livestock or veterinary applications. The term "patient" is intended to mean a human and is therefore used throughout the specification and in the claims. The primary application of the peptides disclosed herein, or peptides having a formula disclosed herein, involves human patients, but the peptides disclosed herein, or peptides having a formula disclosed herein, can be applied to experimental animals, farm animals, zoo animals, wild animals, pets, sport animals, or other animals. Clinical indications and specific uses include the following:

[0142] 2.1 Inflammatory and fibrotic diseases and disorders.

[0143] The peptides and compositions of the present invention (including, but not limited to, peptides that are MC1r, MC3r, MC4r and / or MC5r agonists or partial agonists or any combination thereof) can be used to treat inflammatory diseases and inflammatory conditions in patients. There are many inflammatory diseases and inflammatory conditions that can be treated in this way. In one aspect, inflammatory conditions are caused by diseases 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-Schönlein purpura (HENoch-) Inflammatory disorders include psoriatic arthritis, reactive arthritis, hemochromatosis, hepatitis, Wegener's granulomatosis, vasculitis syndrome, Lyme disease, familial Mediterranean fever, hyperimmunoglobulinemia with periodic fever, TNF receptor-associated periodic syndrome, and inflammatory bowel diseases (including Crohn's disease and ulcerative colitis). On the other hand, inflammatory conditions are caused by diseases including forms of inflammatory bowel diseases such as Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diverticulitis, Behçet's syndrome, infectious colitis, and undifferentiated colitis. On the other hand, inflammatory conditions are caused by autoimmune diseases, including but not limited to systemic syndromes such as systemic lupus erythematosus, Shoglen syndrome, scleroderma, rheumatoid arthritis, and polymyositis, or syndromes that affect only local body systems, such as syndromes affecting the endocrine system (type 1 diabetes, Hashimoto's thyroiditis, Addison's disease, etc.), syndromes affecting the skin system (pemphigus vulgaris), syndromes affecting the blood system (autoimmune hemolytic anemia), or syndromes affecting the nervous system (multiple sclerosis). Therefore, in addition to the general syndromes mentioned above, autoimmune diseases also include the following diseases and conditions: such as acute disseminated encephalomyelitis, Addison's disease, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, autoimmune oophoritis, celiac disease, Crohn's disease, pemphigoid of pregnancy, pulmonary hemorrhage and nephritis syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, idiopathic thrombocytopenic purpura, Kawasaki disease, lupus erythematosus, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, oculoclonus myoclonus syndrome, optic neuritis, Ord thyroiditis, pemphigus, pernicious anemia, primary biliary cirrhosis, Ritter syndrome, Sjogren's syndrome, Takayasu's arteritis, temporal arteritis, autoimmune hemolytic anemia, and Wegener's granulomatosis.

[0144] In another aspect, the inflammatory condition is caused by or associated with chronic obstructive pulmonary disease (COPD), also known as chronic obstructive airway disease, including but not limited to diseases characterized by a pathological limitation (not completely reversible) of airflow in the airways, such as, for example, chronic bronchitis, emphysema, pneumoconiosis, lung tumors, and other pulmonary disorders. Other inflammatory conditions include upper or lower airway diseases and disorders, such as allergic asthma, non-allergic asthma, allergic rhinitis, vasomotor rhinitis, allergic conjunctivitis, non-allergic conjunctivitis, and the like, as well as airway diseases associated with external toxins or substances, such as various forms of pneumoconiosis (coal workers' pneumoconiosis, asbestosis, silicosis, bauxite fibrosis, beryllium poisoning, or siderosis), byssinosis, or hypersensitivity pneumonitis (farmer's lung or bird-fancier's lung). Other pulmonary diseases involving inflammatory conditions include acute respiratory distress syndrome. The peptides and compositions of the application are particularly useful for treating conditions in which glucocorticoids are ineffective or insufficient to produce a desired pharmacological response, such as COPD, asthma in smoking individuals, and other conditions characterized in whole or in part by: accumulation of eosinophils in the lung, neutrophil infiltration and activation, alveolar macrophage recruitment and activation, IL-8 expression by epithelial cells, or increased TNF-α expression. For airway or pulmonary disorders, in one aspect, the peptides of the application are delivered systemically; in another aspect, the peptides of the application are delivered locally, such as by inhalation administration.

[0145] In yet another aspect, the inflammatory condition is caused by or associated with some form of a 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 can also occur in other transplants, and particularly those in which T cells are present in the graft, whether as contaminants or intentionally introduced. Transplantation of body organs (e.g., kidney, liver, pancreas, spleen, uterus, heart, or lung), as well as bone, cornea, face, hand, penis, or skin can all be subject to hyperacute, acute, or chronic rejection. In one embodiment, a pharmaceutical composition including one or more of the peptides of the application is prophylactically administered immediately prior to, during, or immediately after the transplantation of a bodily fluid, organ, or portion to limit or prevent a transplant-related condition or syndrome. In another embodiment, the bodily fluid, organ, or portion to be transplanted is perfused with a solution of a pharmaceutical composition including one or more of the peptides of the application. In yet another embodiment, one or more of the peptides of the application are administered in conjunction with, in combination with, or sequentially with one or more other agents used for transplant rejection, such as a calcineurin inhibitor (including cyclosporine or tacrolimus), an mTOR inhibitor (including sirolimus or everolimus), an anti-proliferative agent (including azathioprine or mycophenolic acid), a corticosteroid (including prednisolone or hydrocortisone), an antibody (such as a monoclonal anti-IL-2Rα receptor antibody, basiliximab or daclizumab, or a polyclonal anti-T cell antibody, such as anti-thymocyte globulin or anti-lymphocyte globulin).

[0146] In yet another aspect, the peptides and compositions of the present application, including but not limited to peptides that are MC1 r, MC3r, MC4r, and / or MC5r agonists or partial agonists or any combination thereof, can be used to treat fibrotic and sclerotic diseases, indications, disorders, and syndromes in a patient. There are a number of fibrotic and sclerotic diseases, indications, disorders, and syndromes that can be so treated. Fibrotic and sclerotic diseases, indications, disorders, and syndromes often include an inflammatory component, and thus many can be similarly classified as inflammatory diseases or disorders and listed above. In addition to including an inflammatory component, fibrotic and sclerotic diseases and disorders can also be idiopathic, toxic, genetic, and / or pharmacologically induced disorders. Generally, 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 MC1 r can lead to inhibition of transforming growth factor-beta 1 -induced collagen synthesis by human dermal fibroblasts, providing therapeutic and / or prophylactic benefit for fibrotic and sclerotic diseases, indications, disorders, and syndromes. Representative fibrotic and sclerotic diseases and disorders that can be so treated include, but are not limited to, localized scleroderma, systemic sclerosis, sclerodermatous graft versus host disease, idiopathic pulmonary fibrosis, bleomycin-induced pulmonary fibrosis, cyclosporine-induced nephropathy, cirrhosis of the liver, hypertrophic scarring, keloids, and the like.

[0147] In yet another aspect, the peptides and compositions of the present application, including but not limited to peptides that are MC1 r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, particularly peptides that are MC1 r and MC5r agonists, can be used to treat fibrotic diseases, disorders, and syndromes in a patient. Such fibrotic processes can be secondary to chronic inflammation, and the development of fibrosis is a common consequence of chronic inflammation. There are a number of diseases in which fibrosis is a cause of death 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 ocular diseases, particularly those characterized by chronic inflammation. It is believed that the peptides and compositions of the present application can both inhibit the formation of fibrosis and have regenerative properties, thereby reducing or mitigating the effects of fibrosis.

[0148] In yet another aspect, the peptides and compositions of the present application, including but not limited to peptides that are MC1 r, MC3 r, MC4 r, and / or MC5 r agonists, partial agonists, antagonists, or any combination thereof, can be used to treat patients for diseases associated with increased cytokine expression, as well as related diseases, indications, conditions, and syndromes. Expression of various cytokines is increased in inflammatory processes, including inflammatory processes secondary to circulatory shock, ischemia, reperfusion injury, and the like. TNF-a is a pleiotropic cytokine produced primarily by macrophages, but also by other types of cells. Other cytokines that are increased in inflammatory processes, including inflammatory processes secondary to circulatory shock, ischemia, reperfusion injury, and the like, include IL-1 and IL-6. While cytokines such as TNF-a have beneficial effects in many situations, significant elevations (such as secondary to circulatory shock, ischemia, reperfusion injury, and the like) can have pathological impacts. In one aspect, reperfusion of hypoxic or ischemic tissue (such as secondary to circulatory shock) results in an inflammatory response, including increased cytokine expression.

[0149] In one embodiment, the present application relates to methods of using one or more of the peptides of the present application to reduce pro-inflammatory cytokine production and expression, including reducing pro-inflammatory cytokine production and expression secondary to circulatory shock, ischemia, reperfusion injury, and the like. Reduction of pro-inflammatory cytokine production and expression, including but not limited to one or more of TNF-a, IL-1, and IL-6, preferably occurs in a short time after administration of a composition comprising one or more of the peptides of the present application.

[0150] In a related embodiment, the present application relates to methods of using one or more of the peptides of the present application and compositions, including but not limited to peptides that are MC1 r, MC3 r, MC4 r, and / or MC5 r agonists, partial agonists, antagonists, or any combination thereof, to increase anti-inflammatory cytokine production and expression. Increase of anti-inflammatory cytokine production and expression, including but not limited to IL-10, occurs in a short time after administration of a composition comprising one or more of the peptides of the present application.

[0151] 2.2 Skin indications.

[0152] In yet another aspect, the peptides and compositions of the present application, including but not limited to peptides that are MC1 r agonists, partial agonists, or antagonists, or combinations thereof, can be used to treat skin and cosmetic diseases, indications, conditions, and syndromes. In one aspect, the peptides and compositions of the present application are MC1 r agonists that stimulate melanocytes and related cells to increase levels of melanin in the skin. By increasing levels of melanin in the skin, protection against ultraviolet radiation (UVR) and sunlight is provided, including protection from skin phototoxicity and photosensitivity caused by UVR, sunlight, and light.

[0153] In yet another aspect, the peptides and compositions of the application, including but not limited to peptides that are MC1 r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be used in the prophylactic and / or therapeutic treatment of skin diseases, indications, disorders, and syndromes, such as acne vulgaris (commonly known as acne), atopic dermatitis (commonly known as atopic eczema or eczema), polymorphous light eruption, psoriasis, rhinophyma, seborrheic dermatitis, vitiligo, porphyria, porphyria cutanea tarda, erythropoietic protoporphyria, solar urticaria, urticarial pigmentosa, or xeroderma pigmentosum. In another aspect, the peptides, compositions, and methods of the application can be used to prevent, limit, or treat photosensitive or light-responsive viral infections, such as herpes simplex virus (commonly known as cold sores and genital herpes depending on the site of infection), human papilloma virus, and varicella zoster virus. In another aspect, the peptides, compositions, and methods of the application can be used to prevent, limit, or treat skin cancers, including for precancerous conditions, and including for actinic keratosis, basal cell carcinoma, melanoma, or squamous cell carcinoma. In another aspect, the peptides, compositions, and methods of the application can be used to prevent or limit adverse effects of various therapies, including light therapies, such as photodynamic therapy. In yet another aspect, the peptides, compositions, and methods of the application can be used to induce tanning, reduce hair graying, or for similar and related purposes involving increased melanin production.

[0154] 2.3 Cancer.

[0155] Certain cancers, such as mesothelioma, are reported to be very sensitive to the pro-growth effects of cytokines and growth factors, and can be treated by peptides selective for MC1 r. Canania, A. et al., “Autocrine inhibitory influences of alpha-melanocyte-stimulating hormone in malignant pleural mesothelioma” J. Leukoc. Biol. 75:253-259 (2004). Cancers that can be treated in this way include pleural mesothelioma (known to express MC1 r and mRHA of the receptor protein), as well as other tumors that express MC1 r, including but not limited to adenocarcinomas, such as lung adenocarcinoma.

[0156] 2.4 Eye diseases and indications.

[0157] There are a number of ocular diseases, indications, conditions, and syndromes characterized by inflammation, including but not limited to increased cytokine production, that can be treated with the peptides and compositions of the present application, including but not limited to peptides that are MC1 r, MC3r, MC4r peptide 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, is an ocular disease that affects approximately 10-20% of the population. This disease affects a greater percentage of the population as age increases, with the majority of these patients being female. In addition, in some cases, such as long visual tasks (e.g., working on a computer), dry environments, use of medications that cause the eyes to dry, etc., ocular irritation sometimes occurs, or symptoms and / or signs of dry eye develop. In individuals with dry eye disease, the protective tear layer that normally protects the ocular surface is compromised, a result of one or more of insufficient or unhealthy tear component production. This can result in exposure of the ocular surface, ultimately promoting drying and damage of the surface cells. Signs and symptoms of dry eye disease include, but are not limited to, keratitis, staining of the conjunctiva and cornea, redness, blurred vision, decreased tear film break-up time, decreased tear production, tear volume, and tear flow, increased conjunctival redness, excess debris in the tear film, ocular dryness, ocular grittiness, ocular burning, ocular foreign body sensation, excessive tearing, photophobia, ocular stinging, refractive defects, ocular sensitivity, and ocular irritation. Patients can experience one or more of these symptoms.

[0158] There are a number of possible variables that can affect the signs or symptoms of dry eye disease in a patient, including circulating hormone levels, various autoimmune diseases (e.g., Sjogren's syndrome and systemic lupus erythematosus), ocular surgery including PRK or LASIK, a number of medications, environmental conditions, visual tasks (e.g., use of a computer), ocular fatigue, wearing contact lenses, and mechanical influences (e.g., corneal sensitivity, partial eyelid closure, surface irregularities (e.g., pterygia), and eyelid irregularities (e.g., ptosis, entropion / ectropion, conjunctival malposition)). Low-humidity environments (e.g., those that result in dehydration) can exacerbate or cause dry eye symptoms, such as sitting in a car with a defroster or living in a dry climate. In addition, visual tasks exacerbate symptoms. Tasks that can greatly affect symptoms include long periods of watching television or using a computer, where blink rates are reduced.

[0159] Uveitis is an ocular disease involving inflammation of the intermediate layer of the eye or uveitis and can 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 occurring in the anterior portion, involving inflammation of the iris and anterior chamber. The condition can occur as a single episode and resolve with appropriate treatment or can have a recurrent or chronic nature. Symptoms include red eye, conjunctival hyperemia, pain, and decreased vision. Signs include dilated ciliary vessels, presence of cells and flare in the anterior chamber, and keratic precipitates on the posterior surface of the cornea. Intermediate uveitis includes inflammation and presence of inflammatory cells in the vitreous cavity, and posterior uveitis includes inflammation of the retina and choroid. Uveitis can be secondary to any of a number of diseases and disorders including acute posterior multifocal placoid pigment epitheliopathy, ankylosing spondylitis, Behcet's disease, birdshot retinochoroidopathy, brucellosis, herpes simplex, herpes zoster, inflammatory bowel disease, juvenile rheumatoid arthritis, Kawasaki disease, leptospirosis, Lyme disease, multiple sclerosis, psoriatic arthritis, Reiter's syndrome, sarcoidosis, syphilis, systemic lupus erythematosus, toxocariasis, toxoplasmosis, tuberculosis, Vogt-Koyanagi-Harada syndrome, Whipple's disease, or Takayasu's arteritis.

[0160] Other ocular inflammatory conditions that can be treated using one or more of the peptides of the present application include, but are not limited to, corneal ulceration, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scarring, epithelial defects, keratoconjunctivitis, idiopathic uveitis, corneal transplantation, age-related macular degeneration, diabetic eye, blepharitis, glaucoma, ocular hypertension, post-operative ocular pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascular membrane, vascular occlusive disease, allergic eye disease, tumors, retinitis pigmentosa, ocular infections, scleritis, ptosis, miosis, ocular pain, mydriasis, neuralgia, cicatricial ocular surface disease, ocular infections, inflammatory eye disease, ocular surface disease, corneal disease, retinal disease, ocular manifestations of systemic disease, inherited ocular disorders, ocular tumors, elevated intraocular pressure, herpes infection, pterygium, wounds persisting to the ocular surface, post-photorefractive keratotomy ocular pain and inflammation, thermal or chemical burns to the cornea, scleral wounds, keratoconus, or conjunctival wounds.

[0161] In one embodiment, the present application relates to methods of treating any of the foregoing ocular diseases, indications, conditions and syndromes using one or more of the peptides of the present application. Such treatment can include treatment by way of delivery of one or more of the peptides of the present application to the ocular surface by eye drops, ointments, gels, lotions, implants, plugs or other means and methods, or alternatively by treatment by way of providing intravitreal injection or similar means for delivery to the vitreous humor, or alternatively by treatment by way of systemic administration to a patient in response thereto, including oral administration, subcutaneous injection or intravenous injection.

[0162] 2.5 Ischemia and Related Indications.

[0163] In yet another aspect, the peptides and compositions of the present application, including but not limited to peptides that are MC1 r, MC3r, MC4r and / or MC5r agonists, partial agonists, antagonists or any combination thereof, can be used to treat ischemia and related diseases, indications, conditions and syndromes. Ischemia includes any reduction or cessation of blood supply to any body organ, tissue, cell or portion, particularly where such reduction or cessation results or can result in ischemic damage to the body organ, tissue, cell or portion. An "ischemic episode" refers to any period of transient or permanent ischemia. Ischemia can result from any constriction or obstruction of the vasculature, or can result from circulatory shock, such as hemorrhagic shock, hypovolemic shock, and the like. The reduction or lack of blood flow results in a reduction or lack of oxygen to the affected body part, and can also result in an increase in inflammatory disease mediators, such as various cytokines and other substances. During certain surgical procedures, such as heart surgery and organ transplantation, blood flow is temporarily stopped and then restored (reperfusion), which results in ischemia-reperfusion injury. During a heart attack, blood supply to the heart is stopped, which also results in ischemia, which in turn evolves into infarction. Current treatment to relieve a heart attack requires reperfusion of the ischemic area of the heart, such as by use of a thrombolytic drug or coronary angioplasty.

[0164] The peptides and compositions of the present application have particular application in preventing injury due to renal ischemia, including lung injury secondary to renal ischemia, preventing or limiting ischemic heart damage following myocardial infarction, preventing or limiting ischemic brain damage following cardiovascular injury, including but not limited to myocardial infarction, stroke, and the like. Neuroprotection is provided by administering the compositions of the present application to a patient suffering from cerebral ischemia or stroke, particularly a patient who is also suffering from hypotension. The peptides and compositions of the present application have particular application in preventing or limiting ischemic organ damage in organ transplantation, including transplantation of the heart, kidney, liver, lung, pancreas or small intestine. In one aspect, the pharmaceutical compositions of the present application can be used for perfusion of a transplanted organ, which can be prior to, during or after organ transplantation.

[0165] In one embodiment, the present application relates to methods of protecting the heart, brain, or other organ of a patient from injury caused by ischemia using one or more of the peptides of the present application. The protective effect against ischemia occurs immediately or within a short time after administration of a composition comprising one or more of the peptides of the present application.

[0166] Ischemia can also be caused by any of a variety of diseases or conditions, and in one embodiment, the present application relates to methods of protecting an organ of a patient from injury caused by ischemia that is caused by a disease or condition using one or more of the peptides of the present application. Such diseases or conditions can include, for example, but are not limited to, atherosclerotic disease (such as atherosclerosis with thrombosis), embolism from the heart or from a blood vessel of any organ, vasospasm, hypotension caused by heart disease, hypotension caused by a systemic disease (including infection or an allergic reaction), or hypotension resulting from administration, ingestion, or other exposure to one or more toxic compounds or drugs. Ischemia can also be secondary ischemia, and in another embodiment, the present application relates to methods of protecting an organ of a patient from injury caused by secondary ischemia using one or more of the peptides of the present application. Such secondary ischemia can be secondary to diseases or conditions such as diabetes, hyperlipidemia, hyperlipoproteinemia, dyslipidemia, Buerger's disease (also known as thromboangiitis obliterans), Takayasu's arteritis, temporal arteritis, Kawasaki's disease (also known as lymphadenopathy syndrome, mucocutaneous lymph node syndrome), infantile polyarteritis, cardiovascular syphilis, and various connective tissue diseases and disorders.

[0167] In yet another aspect, the peptides and compositions of the present application, including but not limited to peptides that are MC1 r, MC3 r, MC4 r, and / or MC5 r agonists, partial agonists, antagonists, or any combination thereof, can be used to treat ischemia-reperfusion injury and related diseases, indications, conditions, and syndromes. While restoration of blood flow after ischemia is essential to protect functional tissue, reperfusion is known to be injurious to tissue in its own right. Both ischemia and reperfusion are known to be important causes of tissue necrosis. Several mechanisms appear to play a pathogenic role in the generation of tissue damage associated with ischemia-reperfusion injury. Certain peptides and compositions of the present application have particular application in preventing or limiting the severity of renal reperfusion injury, including lung injury secondary to renal reperfusion, preventing or limiting reperfusion heart injury following myocardial infarction, preventing or limiting reperfusion brain injury following cardiovascular injury, including but not limited to myocardial infarction, stroke, and the like. The present application has additional particular application in preventing or limiting reperfusion organ damage in organ transplantation, including transplantation of the heart, kidney, liver, lung, pancreas, or small intestine. In one aspect, pharmaceutical compositions of the present application can be used for perfusion of a transplanted organ, which can be prior to, during, or after organ transplantation.

[0168] In one embodiment, the present application relates to methods of protecting the heart, brain, or other organs of a patient from injury caused by ischemia-reperfusion injury (including injury caused by or during reperfusion) using one or more of the peptides of the present application. The protective effect against ischemia-reperfusion injury occurs immediately or within a short time after administration of a composition comprising one or more of the peptides of the present application.

[0169] In yet another aspect, the peptides and compositions of the present application, including but not limited to peptides that are MC1 r, MC3r, MC4r, and / or MC5r agonists, partial agonists, antagonists, or any combination thereof, can be used to treat circulatory shock and related diseases, indications, conditions, and syndromes in a patient. The present application provides peptides, compositions (for use), and methods of treating or preventing shock (including hemorrhagic shock) in a patient, including administering a composition comprising one or more of the peptides of the present application to a patient diagnosed with hemorrhage. Hemorrhage can (but need not) be measured as a percentage of the blood volume of the subject, such as, for example, a loss of greater than about 15% of total blood volume or greater than 20%, 25%, 30%, 35%, 40%, or 50% of the total volume of the subject. Alternatively, hemorrhage can (but need not) be measured according to any amount of blood volume loss sufficient to cause hemorrhagic shock in a particular subject, such as, for example, about 750 mL, 1000 mL, about 1500 mL, or about 2000 mL or more of blood loss in a human subject. Hemorrhage can also be measured according to a decrease in systolic blood pressure, such as, for example, a decrease in systolic blood pressure of about 20 mm Hg, 30 mm Hg, 40 mm Hg, 50 mm Hg, 60 mm Hg, 70 mm Hg, 80 mm Hg, 90 mm Hg, or 100 mm Hg or more than 100 mm Hg below the normal systolic blood pressure of the subject. In particular embodiments, the subject is undergoing or has undergone a medical procedure, such as, but not limited to, surgery, blood transfusion, or childbirth. In other particular embodiments, the subject has suffered a traumatic injury, such as, but not limited to, a traumatic injury resulting from a motor vehicle accident, an industrial injury, or a gunshot wound.

[0170] In further embodiments of the application, the compositions and methods are used to treat cardiogenic shock, hypovolemic shock, and vasodilatory shock, each of which can be at any stage of shock. In a particular embodiment of the application, the methods are used to treat cardiogenic shock. Generally, cardiogenic shock is a low blood flow or perfusion resulting from cardiac dysfunction in which the heart is unable to pump out enough blood. Causes can include any disorder that interferes with ventricular filling or emptying, such as, but not limited to, embolism, ischemia, regurgitation, and valvular dysfunction. In another particular embodiment of the application, the methods are used to treat vasodilatory shock. Vasodilatory shock results from severe dilation of veins or arterioles, which results in insufficient blood flow. Several known causes of vasodilatory shock include, but are not limited to, brain trauma, drug or poison toxicity, anaphylaxis, liver failure, bacteremia, and sepsis. In another more particular embodiment of the application, the methods are used to treat shock resulting from sepsis or bacteremia. In even more particular embodiments, the compositions and methods are used to treat infectious or bacteremic shock in shock, referred to as Stage I, II, or III shock. In yet another embodiment, the compositions and methods of the application are used to treat hypovolemic shock. Generally, hypovolemic shock is a reduction in intravascular volume, where the reduction in intravascular volume can be relative or absolute. Bleeding resulting from a variety of conditions, such as, but not limited to, ulcers, gastrointestinal injury, trauma, accidents, surgery, and aneurysm, can result in hypovolemic shock; but loss of other body fluids can also result in hypovolemic shock. For example, loss of renal fluid, intravascular fluid, water, or other peritoneal fluid can result in hypovolemic shock. In a particular embodiment of the application, the compositions and methods, including administration of one or more of the peptides of the application, are used to treat hypovolemic shock. In even more particular embodiments, the compositions and methods are used to treat Stage I, II, or III hypovolemic shock.

[0171] In one embodiment, the application relates to methods of using one or more of the peptides of the application to protect the heart, brain, or other organ of a patient from injury caused by circulatory shock. The protective effect against circulatory shock occurs immediately or within a short time after administration of a composition comprising one or more of the peptides of the application, preferably within at least about 40 minutes after administration.

[0172] 2.6 MC4r-responsive indications.

[0173] In yet another aspect, the peptides and compositions of the application, including but not limited to peptides that are MC4r agonists or partial agonists or MC3r agonists, partial agonists, antagonists, or any combination thereof, can be used to treat diseases, disorders, and / or conditions that are responsive to modulation of MC4r function, more particularly activation of MC4r (i.e., diseases, disorders, and / or conditions that would benefit from agonism (including full or partial agonism) of MC4r), or diseases, disorders, and / or conditions that are responsive to modulation of MC3r function, more particularly activation of MC3r (i.e., diseases, disorders, and / or conditions that would benefit from agonism (including full or partial agonism) of MC3r), or diseases, disorders, and / or conditions that are responsive to modulation of both MC4r and MC3r function, including energy homeostasis and metabolism-related diseases, disorders, and / or conditions (e.g., diabetes, particularly type 2 diabetes; dyslipidemia; fatty liver; gout; hypercholesterolemia; hypertriglyceridemia; hyperuricemia; impaired glucose tolerance; impaired fasting glucose; insulin resistance syndrome; and metabolic syndrome), food intake-related diseases, disorders, and / or conditions (e.g., excessive eating; binge eating; bulimia; and compulsive eating), and / or energy balance and body weight-related diseases, disorders, and / or conditions, more particularly such diseases, disorders, and conditions characterized by excess body weight and / or excessive food intake. In one aspect, the compounds of the application are used to treat conditions associated with various genetic diseases of expression or receptor, such as proopiomelanocortin deficiency (POMC heterozygous deficiency obesity) due to POMC gene mutation, Prader-Willi syndrome, obesity due to MC4r deficiency, leptin receptor deficiency obesity, leptin deficiency obesity (including congenital leptin deficiency), Bardet-Biedl syndrome, syndrome, and various other diseases, conditions, genetic defects, metabolic disorders, and syndromes.

[0174] In particular, it is believed that such peptides can be used to treat body weight-related diseases, disorders, and / or conditions characterized by excess body weight (including obesity and overweight) (by promoting weight loss, maintaining weight loss, and / or preventing weight gain, including drug-induced weight gain or weight gain following smoking cessation), as well as 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; cardiac disorders (e.g., coronary heart disease, myocardial infarction); cardiovascular disorders; nonalcoholic fatty liver disease (including nonalcoholic steatohepatitis); joint disorders (including secondary osteoarthritis); gastroesophageal reflux; sleep apnea; atherosclerosis; stroke; macro- and microvascular disease; fatty degeneration (e.g., in the liver); gallstones; and gallbladder disorders.

[0175] MC4r is part of the leptin-melanocortin pathway or proopiomelanocortin (POMC)-MC4r pathway. Constituent members of this pathway include a variety of proteins, including a-MSH, POMC, leptin, and the leptin receptor. Certain diseases, disorders, and syndromes are caused by mutations and variations associated with or in one or more constituent members of the POMC-MC4r pathway, including genetic deficiency disorders. As described below, the compounds of the present application can be used to treat diseases, disorders, and syndromes caused by mutations and variations associated with or in one or more constituent members of the POMC-MC4r pathway, including genetic deficiency disorders.

[0176] The hypothalamic POMC-MC4r pathway is part of the regulatory system that modulates feeding behavior, appetite, and body weight. A number of diseases, disorders, and syndromes have been described that are associated with disruption of the hypothalamic POMC-MC4r pathway, believed to be caused by genetic deficiencies or disruptions, including deficiencies or disruptions of genes in the POMC-MC4r pathway. For example, Prader-Willi syndrome is characterized by marked hyperphagia and severe obesity, and can include other features and signs, such as learning disabilities, neurological abnormalities, hypogonadism, short stature, and developmental and cognitive delays. As described below, the compounds of the present application can be used to treat Prader-Willi syndrome as well as other diseases, disorders, and syndromes involving deficiencies or disruptions of genes in the POMC-MC4r pathway.

[0177] Accordingly, the compounds of the present application can be used and are useful in 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 on chromosome 2, position 23.3. POMC gene mutations that result in complete loss or significantly reduced production of the POMC polypeptide result in no a-MSH production or reduced production. This loss of endogenous a-MSH results in significantly reduced MC4r activity, leading to hyperphagia and obesity. The compounds of the present application can be used as surrogate MC4r agonist therapeutic agents for patients with little or no endogenous a-MSH.

[0178] For a variety of diseases, disorders, or syndromes associated with disruption of the hypothalamic POMC-MC4r pathway, various genetic tests and genotyping tests can be employed as part of the diagnosis of a prospective patient to determine the suitability of the use of a compound of the application in such a prospective patient. For example, but not by way of limitation, for Prader-Willi Syndrome, genetic tests (such as DNA-based methylation tests) can be utilized to determine the loss of active genes in a particular portion of chromosome 15 (the 15ql l-q13 region), particularly the loss of at least the paternal copy of the 15ql l-q13 region of chromosome 15. Similarly, POMC deficiency can be diagnosed by a loss-of-function mutation in the POMC gene. Thus, treatment with a compound of the application can include various diagnostic and genetic tests to determine whether there is a loss-of-function mutation or other mutation in the POMC-MC4r pathway, including but not limited to loss-of-function mutations affecting Prader-Willi Syndrome in the 15ql l-q13 region, loss-of-function mutations in the POMC gene, the leptin gene, the leptin receptor gene, and various other genes in the POMC-MC4r pathway.

[0179] In yet another aspect, the peptides and compositions of the application, 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 particular embodiment, the peptides, compositions, and methods of the application are used in male patients to increase erectile function, including but not limited to increasing erectile function to allow vaginal intercourse. In another particular embodiment, the peptides, compositions, and methods of the application are used to treat female sexual dysfunction, including but not limited to increasing arousal success rate, desire success rate, arousal and desire levels. For female sexual dysfunction, including hypoactive sexual desire disorder, endpoints can (but need not) be determined by any of a number of well- validated tools, including but not limited to the Female Sexual Distress Scale, the Female Sexual Encounter Profile, the Female Sexual Function Index, and the Global Assessment Questionnaire. Patients treated for female sexual dysfunction can be premenopausal women or postmenopausal women.

[0180] In yet another aspect, the peptides and compositions of the application, including but not limited to peptides that are MC4r agonists or partial agonists, can be used to inhibit alcohol consumption, or to reduce alcohol consumption, or to treat or prevent alcoholism, or to treat or prevent alcohol abuse, or to treat or prevent alcohol-related disorders. In another related aspect, one or more peptides of the application can be used to inhibit consumption of drugs of abuse, or to reduce consumption of drugs of abuse, or to treat or prevent drug abuse, or to treat or prevent drug abuse-related disorders. Drugs of abuse are typically controlled substances. These controlled substances include controlled naturally-derived drugs (such as heroin, morphine, opium, cocaine, marijuana, etc.), as well as synthetic drugs, such as hydrocodone, methadone, tramadol, various methamphetamine drugs and other known abused tranquilizers, stimulants or depressants, as well as drugs without established medicinal utility, such as Ecstasy, LSD or PCP.

[0181] In yet another aspect, the peptides and compositions of the application, including but not limited to peptides that are MC4r antagonists or optionally inverse agonists of MC4r, including MC4r antagonist or inverse agonist peptides that can be agonists, partial agonists, antagonists or inverse agonists of one or more of MC1 r, MC2r, MC3r and MC5r, can be used to treat a variety of body weight disorders, including cachexia, sarcopenia and wasting syndromes or diseases, as well as to treat inflammation and immune disorders. Body weight disorders include one or more "wasting" disorders (e.g., wasting syndrome, cachexia, sarcopenia) that result in undesirable and unhealthy weight loss or reduction in body cell mass. In the elderly, as well as in cancer and AIDS patients, wasting diseases can result in undesirable weight loss, including both fat and non-fat compartments. Wasting diseases can be the result of insufficient food intake and / or metabolic changes associated with the disease and / or the aging process. Cancer patients and AIDS patients, as well as patients following extensive surgery or suffering from chronic infections, immune diseases, hyperthyroidism, Crohn's disease, psychogenic disease, chronic heart failure or other serious trauma, often suffer from wasting diseases. Wasting diseases are sometimes also referred to as cachexia, and are generally considered to be metabolic disorders, as well as sometimes eating disorders. In addition, cachexia can be characterized by high metabolism and catabolism. Typically, sarcopenia, another such disorder that affects aging individuals, is characterized by a reduction in muscle mass. Terminal stage wasting diseases, as described above, can develop in individuals suffering from cachexia or sarcopenia.

[0182] 2.7 Nuclear medicine and drug delivery applications.

[0183] In yet another aspect, the peptides and compositions of the application, including but not limited to peptides that are MClr agonists, partial agonists, or antagonists, can be used for targeted imaging and cytotoxic therapy of certain cancers, such as melanoma, and other indications in patients in need thereof. The peptides, compositions, and methods of the application can be used for imaging melanoma and other cancers or diseases or conditions that are characterized in part by relatively high expression of MClr, such as by diagnostic imaging using a radionuclide in combination with a peptide of the application. For diagnostic imaging, a peptide of the application is typically conjugated to a radionuclide by use of a linker, such as a crosslinker that couples a peptide of the application to a 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 99m Tc, 111 In, 123 I, and 67 Ga, and the like. Positron emitters that can be used in this way include 11 C, 13 N, 15 O, and 18 F.

[0184] In a related aspect, the peptides, compositions, and methods of the application can be used for cytotoxic therapy of melanoma, other cancers or diseases or conditions that are characterized in part by relatively high expression of MClr, such as by use of a chemotherapeutic agent, including a toxin or a radiotherapeutic agent, in combination with a peptide of the application. Chemotherapeutic agents include any antineoplastic drug or chemical, such as, for example, alkylating agents, antimetabolites, anthracyclines, plant alkaloids, topoisomerase inhibitors, and other antineoplastic agents. Non-limiting examples of alkylating agents include cisplatin, carboplatin, oxaliplatin, mechlorethamine, cyclophosphamide, chlorambucil, and ifosfamide; 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 topoisomerases (such as amsacrine, etoposide, etoposide phosphate, and teniposide). However, any agent suitable for use in targeted cytotoxic therapy can be used in this way. Non-limiting examples of radiotherapeutic agents that can be used in this way include 131 I, 125 I, 211 At, 186 Re, 188 Re, 90 Y, 153 Sm,212 Bi and 32 P, etc.

[0185] Diagnostic imaging or cytotoxic therapy agents can be incorporated into the peptides of the present application (e.g., as by use of a 11 C, 13 N, 15 O, etc. in place of a non-radioactive isotope); can be attached directly to the peptides of the present application (as, for example, by halogenation or other direct complexation methods); or can be attached indirectly to the peptides of the present application, such as by way of a linker or chelating unit. Linker units are well known in the art and include, but are not limited to, chemically linked conjugates that include at least one disulfide bond, thioether bond, or covalent bond between free reactive groups. Representative cross-linking and conjugation reagents are disclosed in U.S. Patent Nos. 7,169,603; 7,820,164; and 5,443,816; and U.S. Publication No. 2009 / 0297444, among others, which are incorporated herein by reference.

[0186] 3.0 Combination therapy for certain indications.

[0187] The peptides, compositions, and methods of the present application can be used to treat any of the foregoing diseases, indications, conditions, or syndromes, or any MC1 r-mediated or responsive disease, indication, condition, or syndrome, by administration in combination with one or more other pharmaceutically active compounds. Such administration in combination can be by way of a single dosage form that includes both the peptides of the present application and one or more other pharmaceutically active compounds, such single dosage forms including tablets, capsules, sprays, inhalable powders, injectable liquids, and the like. Alternatively, administration in combination can be by way of administration of two different dosage forms, with one dosage form containing the peptides of the present application and the other dosage form including another pharmaceutically active compound. In this case, the dosage forms can be the same or different. The term "co-administration" indicates that each of the at least two compounds in a combination therapy is administered within a time frame in which the respective biological activities or effects overlap. Thus, the term includes both consecutive administration and simultaneous administration of the compounds, with one of the compounds being one or more of the peptides of the present application. If more than one compound is co-administered, the routes of administration of the two or more compounds need not be the same. Without intending to limit combination therapy, certain combination therapies that can be used are illustrated below.

[0188] 3.1 Combination therapy with anti-inflammatory agents.

[0189] For the treatment of diseases, indications, conditions and syndromes associated with inflammation, the peptides of the present application can be used in combination therapy, including by way of co-administration with one or more anti-inflammatory agents. One class of anti-inflammatory agents is the glucocorticoids, including but not limited to cortisone, including cortisone acetate, hydrocortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, prednisone, flurandrenolide, deoxycorticosterone acetate, and aldosterone. Other anti-inflammatory agents that can be used in combination therapy, including by way of co-administration, include aspirin, non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen and naproxen, TNF-α inhibitors such as tenidap and rapamycin or derivatives thereof, or TNF-α antagonists (e.g., infliximab, OR1384), cyclooxygenase inhibitors (i.e., COX-1 and / or COX-2 inhibitors such as or ), CTLA4-lg agonists / antagonists, CD40 ligand antagonists, IMPDH inhibitors (e.g., mycophenolate mofetil ( )), integrin antagonists, alpha-4 beta-7 integrin antagonists, cell adhesion inhibitors, interferon gamma antagonists, ICAM-1, prostaglandin synthesis inhibitors, budesonide, clofazimine, p38 mitogen-activated protein kinase inhibitors, protein tyrosine kinase (PTK) inhibitors, IKK inhibitors, therapies for irritable bowel syndrome (e.g., and Maxi- Kane® openers such as those disclosed in U.S. Patent No. 6,184,231), or other NF-κB inhibitors such as corticosteroids, calphostin, CSAIDs, 4-substituted imidazo[l,2-A]quinoxalines (as disclosed in U.S. Patent No. 4,200,750); interleukin-10, salicylates, nitric oxide, and other immunosuppressants; and nuclear translocation inhibitors such as deoxyspergualin (DSG).

[0190] 3.2 Combination therapy with phosphodiesterase inhibitors.

[0191] For certain applications and indications, it is desirable to increase and maintain levels of cyclic adenosine 3', 5' monophosphate (cAMP), a nucleotide messenger associated with inflammatory cell activity. The peptides of the present invention increase intracellular levels of cAMP and can be co-administered with compounds or agents that inhibit cAMP degradation. cAMP is hydrolyzed to an inactive form by phosphodiesterases (PDEs); thus, compounds or agents that inhibit PDEs can result in the maintenance and / or increase in the availability of cAMP. One class of compounds, known as PDE inhibitors, has been extensively studied for the treatment of inflammatory diseases, such as asthma, COPD, and acute respiratory distress syndrome. Preferred are inhibitors of PDE types 1, 2, 3, 4, 7, 8, 10, or 11; in one aspect, this includes cAMP-PDE inhibitors that are selective 4-type PDE inhibitors or inhibitors selective for one particular type of PDE 4 isozyme, such as, for example, rolipram, cilomilast, ibudilast, and pirazolamast.

[0192] 3.3 Combination therapy for ocular indications.

[0193] For ocular indications, ophthalmic dosage forms can include one or more active ingredients in addition to one or more of the peptides of the present invention, such as, for example, artificial tear ingredients, topical corticosteroids, non-steroidal anti-inflammatory drugs, or calcineurin inhibitors, such as cyclosporin-A ophthalmic emulsion (RESTASIS® (Allergan)). Co-administration can also include administration of one or more additional compounds administered separately from the peptides of the present invention, such as ophthalmic dosage forms administered separately including artificial tear ingredients, topical corticosteroids, non-steroidal anti-inflammatory drugs, calcineurin inhibitors (e.g., cyclosporin-A), or combinations of any of the foregoing drugs.

[0194] Combination ophthalmic solutions can be used, particularly including solutions comprising more than one active pharmaceutical ingredient. In one aspect, a non-steroidal anti-inflammatory drug (NSAID) is used in combination with the peptides of the present application. NSAIDs suitable for use in combination ophthalmic solutions include agents that inhibit the cyclooxygenase (COX)-1 and / or cyclooxygenase-2 enzymes, their esters, and pharmaceutically acceptable salts thereof (including, but not limited to, propionic acid compounds, such as naproxen, flurbiprofen, oxaprozin, ibuprofen, ketoprofen, fenoprofen; ketorolac tromethamine; acetic acid derivatives, such as sulindac, indomethacin, and etodolac; benzoic acid, such as diclofenac, bromfenac, and suprofen; aryl acetic acid prodrugs, such as nepafenac and amfenac; salicylic acid, such as aspirin, salsalate, diflunisal, trisalicylate choline magnesium; p-aminophenol derivatives, such as acetaminophen; naphthylalkanone, such as nambumetone; enolic acid derivatives, such as piroxicam and meloxicam; femanate, such as mefenamic acid, meclofenamic acid, and flufenamic acid; pyrrole acetic acid, such as tolmetin; and pyrazolone, such as phenylbutazone); and COX-2 selective inhibitors, such as celecoxib, valdecoxib, parecoxib, etoricoxib, and luaricoxib. Ophthalmic solutions can additionally comprise other active ingredients, including, but not limited to, vasoconstrictors, anti-allergic agents, anti-infective agents, steroids, anesthetics, anti-inflammatory agents, analgesics, dry eye treatments (e.g., secretagogues, mucomimetics, polymers, lipids, antioxidants), etc., or can be administered (simultaneously or sequentially) in combination with pharmaceutical compositions comprising other active ingredients, including, but not limited to, vasoconstrictors, anti-allergic agents, anti-infective agents, steroids, anesthetics, anti-inflammatory agents, analgesics, dry eye treatments (e.g., secretagogues, mucomimetics, polymers, lipids, antioxidants), etc.

[0195] 3.4 Combination therapy for shock-related indications.

[0196] In addition to one or more of the peptides of the present application, the methods of the present application for treating or preventing circulatory shock involve co-administering one or more substances to the subject. For example, one or more of the peptides of the present application can be co-administered with androstenetriol, androstenediol or a derivative thereof, various vasopressin agonists or other pharmaceutically active substances such as a catecholamine or other alpha adrenergic agonist, alpha2 adrenergic agonist, beta adrenergic agonist or beta2 adrenergic agonist, including but not limited to epinephrine, norepinephrine, dopamine, isoproterenol, vasopressin and dobutamine. Alternatively, one or more of the peptides of the present application can be co-administered with a fluid or other substance capable of alleviating, attenuating, preventing or eliminating symptoms in a subject suffering from, exhibiting symptoms of 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 of the peptides of the present application is specific to the particular subject suffering from, exhibiting symptoms of or at risk of suffering from shock. For example, fluids that can be co-administered with one or more of the peptides of the present application include, but are not limited to, salt solutions such as sodium chloride and sodium bicarbonate as well as whole blood, synthetic blood substitutes, plasma, serum, serum albumin and colloidal solutions. Colloidal solutions include, but are not limited to, solutions containing hydroxyethyl starch, albumin or plasma. In a particular embodiment of the present application, a fluid such as one or more of a salt solution, a colloidal solution, whole blood, a synthetic blood substitute, plasma or serum is co-administered with one or more of the peptides of the present application to a patient suffering from or exhibiting symptoms of hypovolemic shock such as hemorrhagic shock.

[0197] 3.5 Combination therapy for obesity and related metabolic syndromes.

[0198] One or more of the peptides of the present application can 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 overweight, in particular other anti-obesity drugs affecting energy expenditure, glycolysis, gluconeogenesis, glycogenolysis, lipolysis, lipogenesis, fat absorption, fat storage, fat excretion, hunger and / or satiety and / or appetite mechanisms, appetite / drive, food intake or gastrointestinal motility. Medications that reduce energy intake include various pharmacological agents known as anorectic drugs, which are used as adjuncts to behavioral therapy in weight loss programs.

[0199] In general, the total dosage of the following obesity-controlling agents or drugs, when used in combination with one or more peptides of the present application, can range from 0.01 to 3,000 mg / day, preferably from about 0.1 to 50 mg / day and more preferably from about 0.1 to 10 mg / day, in single or 2-4 divided doses. The exact dosage, however, is determined by the attending clinician and is dependent on a number of factors including the potency of the compound administered, the age, weight, condition and response of the patient.

[0200] One or more peptides of the present application can be combined with one or more other pharmacologically active agents useful in the treatment of diabetes, such as other anti-diabetic agents.

[0201] One or more peptides of the present application can additionally or alternatively be further combined with one or more other pharmacologically active agents useful in the treatment of diseases, disorders and / or conditions associated with obesity and / or being overweight, 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 (including non-alcoholic steatohepatitis); joint disorders (including secondary osteoarthritis); gastroesophageal reflux; sleep apnea; atherosclerosis; stroke; macro- and microvascular diseases; fatty degeneration (e.g., in the liver); gallstones; and gallbladder disorders.

[0202] According to another aspect of the present application, there is provided a combination therapy comprising administering to a mammal (e.g., a human) in need of such therapeutic treatment a pharmacologically effective amount of a peptide according to the present application, or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable diluent or carrier, and simultaneously, sequentially or separately administering one or more of the following agents:

[0203] - insulin and insulin analogs;

[0204] - insulin secretagogues, including sulfonylureas (e.g., glyclazide) and prandial glucose regulators (sometimes referred to as "short-acting secretagogues"), such as meglitinides (e.g., repaglinide and nateglinide);

[0205] - agents that improve the action of incretins, 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);

[0206] - insulin sensitizers, including peroxisome proliferator-activated receptor gamma (PPAR gamma) agonists, such as thiazolidinediones (e.g., pioglitazone and rosiglitazone), and agents having activity at any combination of PPAR alpha, gamma and delta;

[0207] - agents that modulate hepatic glucose balance, such as biguanides (e.g., metformin), fructose 1,6-bisphosphatase inhibitors, glycogen phosphorylase inhibitors, glycogen synthase kinase inhibitors, and glucokinase activators;

[0208] - agents designed to reduce / slow down intestinal glucose absorption, such as a-glucosidase inhibitors (e.g., miglitol and acarbose);

[0209] - agents that antagonize the action of glucagon or reduce its secretion, such as amylin analogs (e.g., pramlintide);

[0210] - agents that prevent the reabsorption of glucose by the kidneys, such as sodium-dependent glucose transporter 2 (SGLT-2) inhibitors (e.g., dapagliflozin);

[0211] - agents designed to treat long-term hyperglycemia complications, such as aldose reductase inhibitors (e.g., epalrestat and ranziostat); and agents for treating complications associated with microangiopathy;

[0212] - anti-dyslipidemic agents, such as HMG-CoA reductase inhibitors (statins, e.g., rosuvastatin) and other cholesterol-lowering agents; PPARa agonists (fibrates, e.g., gemfibrozil and fenofibrate); bile acid sequestrants (e.g., cholestyramine); inhibitors of cholesterol absorption (e.g., phytosterols (i.e., plant sterols), synthetic inhibitors); cholesteryl ester transfer protein (CETP) inhibitors; inhibitors of the ileal bile acid transport system (IBAT inhibitors); bile acid binding resins; niacin (nicotinic acid) and analogs thereof; antioxidants, such as probucol; and omega-3 fatty acids;

[0213] - antihypertensive agents, 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 dihydropyridines (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); central adrenergic agents, such as central alpha agonists (e.g., clonidine); and diuretics (e.g., furosemide);

[0214] - hemostasis modulators, including anti-thrombotics, such as activators of fibrinolysis; thrombin antagonists; factor Vila inhibitors; anticoagulants, such as vitamin K antagonists (e.g. warfarin), heparin and low-molecular-weight analogs thereof, factor Xa inhibitors and direct thrombin inhibitors (e.g. argatroban); anti-platelet 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);

[0215] - anti-obesity agents, such as anorectic agents (e.g. ephedrine) including noradrenergic agents (e.g. phentermine) and serotoninergic activation 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);

[0216] - eating behaviors modulators, such as orexin receptor modulators and melanin-concentrating hormone (MCH) modulators;

[0217] - glucagon-like peptide-1 (GLP-1) receptor modulators;

[0218] - neuropeptide Y (NPY) / NPY receptor modulators;

[0219] - pyruvate dehydrogenase kinase (PDK) modulators;

[0220] - serotonin receptor modulators;

[0221] - leptin / leptin receptor modulators;

[0222] - ghrelin / ghrelin receptor modulators; or

[0223] - monoamine transmission modulators, such as selective serotonin reuptake inhibitors (SSRIs) (e.g. fluoxetine), norepinephrine reuptake inhibitors (NARIs), norepinephrine-serotonin reuptake inhibitors (SNRIs), triple monoamine reuptake blockers (e.g. tesofensine) and monoamine oxidase inhibitors (MAOIs) (e.g. toloxinone and amiflamine),

[0224] or pharmaceutically acceptable salts, solvates, solvates of such salts or prodrugs thereof, optionally together with a pharmaceutically acceptable carrier.

[0225] According to another aspect of the application, there is provided a combination therapy comprising administration of a pharmacologically effective amount of a compound according to the application, or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable carrier, and simultaneous, sequential or separate administration of a very low calorie diet (VLCD) or a low calorie diet (LCD).

[0226] According to still another aspect of the application, one or more peptides of the application, and preferably a peptide that is an MC4r agonist, can be administered in combination with a GLP-1 receptor agonist, as disclosed in WO 2016 / 168388 "Therapies for Obesity, Diabetes and Related Indications", which is incorporated herein by reference. Thus, the present application includes a pharmaceutical composition for subcutaneous administration for the treatment of obesity or for inducing weight loss, which comprises, on a per dose basis:

[0227] a peptide of the application that is an MC4r agonist, in an amount that is sufficient to induce at least minimal weight loss when administered as a monotherapy not in combination with a GLP-1 receptor agonist; and

[0228] a GLP-1 receptor agonist, in an amount that is sufficient to induce glycemic control but not weight loss when administered as a monotherapy not in combination with an MC4r agonist, wherein the pharmaceutical composition preferably has a synergistic anti-obesity effect.

[0229] In a related aspect, the present application provides a method of treating a patient suffering from obesity, diabetes or metabolic syndrome, comprising administering to the patient (a) a peptide of the application that is an MC4r agonist, in an amount that is sufficient to induce at least minimal weight loss when administered as a monotherapy not in combination with a GLP-1 receptor agonist; and (b) a GLP-1 receptor agonist, in an amount that is sufficient to induce glycemic control but not weight loss when administered as a monotherapy not in combination with an MC4r agonist. Preferably, the method results in a synergistic effect on obesity treatment.

[0230] In another aspect, the present application provides a method of reducing side effects associated with a therapeutic agent for treating obesity, diabetes or metabolic syndrome in a patient, comprising:

[0231] administering an amount of a peptide of the application that is an MC4r agonist, wherein the amount of MC4r agonist peptide administered would not be sufficient to elicit a desired pharmacological response in treating at least one condition from the group comprising obesity, diabetes and metabolic syndrome in a patient when administered as a monotherapy not in combination with a GLP-1 receptor agonist; and

[0232] administering an amount of a GLP-1 receptor agonist, wherein the amount of GLP-1 receptor agonist administered is insufficient to elicit a desired pharmacological response in treating at least one condition from the group comprising obesity, diabetes, and metabolic syndrome when administered as a monotherapy not in combination with a MC4r agonist;

[0233] wherein the amount of MC4r agonist and the amount of GLP-1 receptor agonist together are effective to elicit a desired pharmacological response in treating at least one condition from the group comprising obesity, diabetes, and metabolic syndrome,

[0234] thereby reducing side effects in treating at least one of obesity, diabetes, or metabolic syndrome.

[0235] 3.6 Combination therapy for sexual dysfunction.

[0236] The cyclic peptides of the present application can also and are contemplated to be used in combination with other drugs or agents, such as for the treatment of sexual dysfunction. These other drugs and agents can include agents that induce erectile activity, including phosphodiesterase 5 (PDE-5) inhibitors, testosterone, prostaglandins, and the like. In a preferred embodiment of the present application, the cyclic peptides of the present application are used in combination with a therapeutically effective amount of a cyclic GMP-specific phosphodiesterase inhibitor or an alpha-adrenergic receptor antagonist. The teachings and disclosures of U.S. Patent No. 7,235,625 entitled "Multiple Agent Therapy for Sexual Dysfunction" are incorporated by reference as if fully set forth herein.

[0237] Accordingly, the present application provides methods of treating sexual dysfunction, the methods comprising the step of administering to a patient suffering from or at risk of suffering from sexual dysfunction a therapeutically effective amount of a cyclic peptide of the present application in combination with a therapeutically effective amount of a second sexual dysfunction pharmaceutical agent. The cyclic peptide of the present application can be administered simultaneously with, prior to, or after administration of a therapeutically effective amount of the second sexual dysfunction pharmaceutical agent. Preferably, the peptide of the present application is administered within one hour, preferably less than half an hour, of administration of a therapeutically effective amount of the second sexual dysfunction pharmaceutical agent. However, for certain forms of combination therapy, such as for example, in combination with a therapeutically effective amount of a hormone or hormone-related dysfunction pharmaceutical agent, the hormone or hormone-related dysfunction pharmaceutical agent can be administered on an independent schedule, such that there is no set or specific temporal relationship between administration of the peptide of the present application and the hormone or hormone-related dysfunction pharmaceutical agent. Thus, for example, the hormone or hormone-related dysfunction pharmaceutical agent can be administered in a daily dose or other dose, or by way of a patch or other continuous administration schedule, with the peptide of the present application being administered when the patient desires or needs.

[0238] Accordingly, the present application provides methods of treating sexual dysfunction, the methods comprising the step of administering to a patient suffering from or at risk of suffering from sexual dysfunction a therapeutically effective amount of a cyclic peptide of the present application in combination with a therapeutically effective amount of a second sexual dysfunction pharmaceutical agent. The cyclic peptide of the present application can be administered simultaneously with, prior to, or after administration of a therapeutically effective amount of the second sexual dysfunction pharmaceutical agent. Preferably, the peptide of the present application is administered within one hour, preferably less than half an hour, of administration of a therapeutically effective amount of the second sexual dysfunction pharmaceutical agent. However, for certain forms of combination therapy, such as for example, in combination with a therapeutically effective amount of a hormone or hormone-related dysfunction pharmaceutical agent, the hormone or hormone-related dysfunction pharmaceutical agent can be administered on an independent schedule, such that there is no set or specific temporal relationship between administration of the peptide of the present application and the hormone or hormone-related dysfunction pharmaceutical agent. Thus, for example, the hormone or hormone-related dysfunction pharmaceutical agent can be administered in a daily dose or other dose, or by way of a patch or other continuous administration schedule, with the peptide of the present application being administered when the patient desires or needs.

[0239] The present application also provides pharmaceutical compositions comprising a cyclic peptide of the present application and a second compound useful in the treatment of sexual dysfunction. In embodiments of the composition, the additional compound useful in the treatment of sexual dysfunction is preferably selected from, but not limited to, the group consisting of: phosphodiesterase inhibitors; cyclic GMP-specific phosphodiesterase inhibitors; prostaglandins; apomorphine; oxytocin modulators; alpha-adrenergic antagonists; androgens; selective androgen receptor modulators (SARMs); bupropion; vasoactive intestinal peptide (VIP); neutral endopeptidase inhibitors (NEP); and neuropeptide Y receptor antagonists (NPY).

[0240] In embodiments of the methods and compositions, the second sexual dysfunction pharmaceutical agent is testosterone.

[0241] In another embodiment of the combination therapy, the second sexual dysfunction pharmaceutical agent is a phosphodiesterase type V (PDE-5) inhibitor. For example, the PDE-5 inhibitor can be Viagra® (the brand name for sildenafil citrate), Viagra® (the brand name for sildenafil citrate), (Cialis®). Other PDE-5 inhibitors are disclosed 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.

[0242] In another embodiment of the above compositions, the compound useful in the treatment of sexual dysfunction is an estrogen agonist / antagonist. In one embodiment, the estrogen agonist / antagonist is (-)-cis-6-phenyl-5-[-4-(2-pyrrolidin-l-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalen-2-ol (also known as lasofoxifene) or an optical isomer or geometric isomer thereof; a pharmaceutically acceptable salt, N-oxide, ester, quaternary ammonium salt; or prodrug thereof. More preferably, the estrogen agonist / antagonist is in the form of a D-tartrate salt.

[0243] In yet another embodiment of the above compositions, 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-l-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, GW 5368, GW 7604, TSE-424, and optical isomers or geometric isomers thereof; and pharmaceutically acceptable salts, N-oxides, esters, quaternary ammonium salts, and prodrugs thereof.

[0244] In yet another embodiment, the cyclic peptides of the present application can be used in combination with any known mechanical aids or devices.

[0245] 4.0 Methods of Administration and Use

[0246] Methods of administration and use vary depending on the particular peptide disclosed herein or peptide having a formula disclosed herein, the disease, indication, disorder, or syndrome to be treated, and other factors known to those of skill in the art. In general, any method of administration and use known in the art or later developed can be used with the peptides disclosed herein or peptides having a formula disclosed herein. Without limiting the foregoing, the following methods of administration and use have particular application to the indicated indications.

[0247] 4.1 Subcutaneous Injection Use

[0248] In one aspect, the composition comprising one or more peptides of the application is formulated for subcutaneous injection and administered subcutaneously at specified intervals, such as once a week or once or multiple times a day. In another aspect, the composition is formulated as an injectable sustained release formulation. In one embodiment, the peptides of the application are formulated 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 salt, polysorbate 80, sodium hydroxide or hydrochloric acid to adjust pH, and the like. In another embodiment, the peptides of the application are formulated with a poly(ortho ester), which can be a self-catalyzing poly(ortho ester) with any variable percentage of lactic acid in the polymer backbone, and optionally one or more additional excipients. In one embodiment, poly(D,L-lactide-co-glycolide) polymers (PLGA polymers) are used, preferably PLGA polymers with hydrophilic end groups, such as PLGA RG502H from Boehringer Ingelheim, Inc. (Ingelheim, Germany). Such formulations can be prepared, for example, by combining the peptides of the application in a suitable solvent (such as methanol) with a solution of PLGA in dichloromethane, and adding to this a continuous phase solution of polyvinyl alcohol in a reactor under suitable mixing conditions. In general, any of a number of injectable and biodegradable polymers, preferably also viscous polymers, can be used in the sustained release injectable formulation. The teachings of U.S. Patent Nos. 4,938,763, 6,432,438, and 6,673,767, and the biodegradable polymers and formulation methods disclosed therein, are incorporated herein by reference. Depending on the concentration and amount of peptide, the rate of biodegradation of the polymer, and other factors known to those skilled in the art, the formulation can be such that injections are required on a weekly, monthly, or other periodic basis.

[0249] 4.2 Inhalation use.

[0250] In one aspect, compositions comprising one or more peptides of the present invention are formulated for administration to the respiratory tract, such as in the form of an aerosol or solution for a nebulizer, or as a fine powder for inhalation or blowing (e.g., topically to the lungs and / or airways), alone or in combination with one or more inert carriers or other active pharmaceutical ingredients, and in the form of solutions, suspensions, aerosols, or dry powder formulations. See also Cryan, S.-A., “Carrier-based strategies for targeting protein and peptide drugs to the lungs,” The AAPS Journal 7:E20-41 (2005). Generally, the peptides of the present invention can be used in 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 No. 20090241949, “Dry powder inhalation system”; U.S. Patent Application No. 20080066741, “Methods and systems of delivering medication via inhalation”; U.S. Patent Application No. 20070298116, “Amorphous, spray-dried powders having a reduced moisture content and a high long-term stability”; U.S. Patent Application No. 20070140976, “Aqueous inhalation pharmaceutical composition”; U.S. Patent Application No. 20060054166, “Inhalation nebulizer”; U.S. Patent Application No. 20050211244, “Dry powder”. Preparations; U.S. Patent Application No. 20050123509. "Modulating charge density to produce improvements in the characteristics of spray-dried proteins";U.S. Patent Application 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”; U.S. Patent No. 7,387,794, “Preparation of powder agglomerate”; U.S. Patent No. 7,258,873, “Preservation of bioactive materials by spray drying”; U.S. Patent No. 7,186,401, “Dry powder for inhalation”; U.S. Patent No. 7,143,764, “Inhalation device”; U.S. Patent No. 7,022,311, “Powderyinhalational preparations and process for producing the US Patent No. 6,962,151, "Inhalation nebulizer"; US Patent No. 6,907,880, "Inhalation device"; US Patent No. 6,881,398, "Therapeutic dry powder preparation"; US Patent No. 6,698,425, "Powder inhaler"; US Patent No. 6,655,380, "Inhalation device"; US Patent No. 6,645,466, "Dry powder for inhalation"; US Patent No. 6,632,456, "Compositions for inhalation"; US Patent No. 6,610,272, "Medicinalaerosol formulation"; US Patent No. 6,596,261, "Method" of administering a medicinal aerosol formulation;US Patent No. 6,585,957, "Medicinal aerosol formulation"; US Patent No. 6,582,729, "Powered pharmaceutical formulations having improved dispersibility"; US Patent No. 6,572,893, "Systems and processes for spray drying hydrophobic drugs with hydrophilic excipients"; US Patent No. 6,551,578, "Modulated release particles for aerosol delivery"; US Patent No. 6,520,179, "Inhalation device"; US Patent No. 6,518,239, "Dry powder compositions having improved dispersivity"; US Patent No. 6,503,481, "Compositions for aerosolization" andinhalation; U.S. Patent No. 6,358,530, "Powered pharmaceutical formulations having improved dispersibility"; U.S. Patent No. 6,325,061, "Inhalation device"; U.S. Patent No. 6,257,232, "Inhalation device"; U.S. Patent No. 6,187,344, "Powered pharmaceutical formulations having improved dispersibility"; U.S. Patent No. 6,116,237, "Methods of dry powder inhalation";U.S. Patent No. 5,934,272, "Device and method of creating aerosolized mist of respiratory drug"; and U.S. Patent No. 5,558,085, "Intrapulmonary delivery of peptide drugs".

[0251] The composition may be a dry powder composition for local delivery to the lungs via inhalation. The composition may be an inhalation powder mixture containing the peptides of the present invention and a suitable powder matrix, 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 reservoir dry powder inhalers, multi-dose dry powder inhalers, or metered-dose inhalers. The composition may include additional excipients such as alcohols, surfactants, lubricants, antioxidants, or stabilizers. Suitable propellants include hydrocarbons, chlorofluorocarbons, and hydrofluorocarbons, or mixtures of any such propellants.

[0252] Inhalation solutions can also be formulated in liquefied propellants for aerosol delivery, such as with pressurized metered-dose inhalers. In yet another formulation, with or without suitable pH or tension regulation, the solution can be in the form of an atomized aqueous suspension or aqueous solution as a single- or multi-dose device.

[0253] 4.3 Nasal delivery.

[0254] Formulations or compositions suitable for nasal application (where the carrier is a solid) include coarse powder having a particle size range of, for example, 20 to 500 micrometers, which is applied by means of nasal fumigation (i.e., rapid inhalation from a powder container near the nose through the nasal passage). For example, suitable powder compositions comprise a powdered formulation of the active ingredient thoroughly mixed with lactose or other acceptable inert powders for intrabronchial application. The powder composition can be applied via an aerosol dispenser or encapsulated in a fragile capsule into which the patient inserts a device that punctures the capsule and expels the powder in a stable stream suitable for inhalation. Alternatively, suitable formulations may contain a liquid carrier, such as, for example, nasal sprays or drops, which may contain an aqueous or oil solution of the active ingredient.

[0255] 4.4 Delivery via the buccal route and via the mucosa.

[0256] The pharmaceutical composition may additionally comprise one or more of the following: water, buffer solution (e.g., neutral buffered saline or phosphate buffered saline), ethanol, mineral oil, vegetable oil, dimethyl sulfoxide, carbohydrates (e.g., glucose, mannose, sucrose, or dextran), mannitol, protein, adjuvant, peptide or amino acid (such as glycine), antioxidant, chelating agent (such as EDTA or glutathione), and / or preservative. Furthermore, one or more other active ingredients may (but need not) be included in the pharmaceutical composition provided herein.

[0257] 4.5 Oral delivery.

[0258] In one aspect, the peptide of the present invention, comprising an MC1r agonist, is orally administered and substantially completely delivered to the lumen of the entire or part of the intestine (in some respects, including the patient's colon) for the treatment of inflammatory bowel disease, colitis, or other melanocortin receptor-mediated or responsive gastrointestinal disorders, indications, symptoms, and syndromes. Delayed-release polymer formulations comprising the peptide of the present invention may be used, including but not limited to pH-dependent release polymers. The teachings and disclosures of International Publication No. WO 2019 / 183472, filed under International Application No. PCT / US2019 / 023575 and entitled “Melanocortin Receptor-Specific Formulations and Methods for Gastrointestinal Tract-Specific Delivery,” are incorporated herein by reference as fully elucidated.

[0259] For systemic administration, compositions comprising one or more peptides disclosed herein or peptides having the formula disclosed herein may be administered orally in a separate dosage form (e.g., tablets or capsules). In a preferred aspect, the separate dosage form includes an enteric coating and optionally one or more agents that increase uptake, reduce protease degradation, increase cell permeability, etc. Any of a variety of delivery technologies (including, but not limited to, liposome compositions, mucosal adhesion or gastric retention delivery systems, absorption enhancers, multifunctional drug delivery systems, co-administration of permeability enhancers and / or protease inhibitors, covalent conjugation with various chemical or biological adjuvants such as to increase cell penetration, enteric coatings, various nanoparticles, etc.) may be used for the oral delivery of the peptides of the present invention.

[0260] 5.0 Preparation method.

[0261] Generally, the peptides disclosed herein, or peptides having the formula disclosed herein, can be synthesized by any means known in the art (including by solid-phase synthesis) and can be purified according to methods known in the art. The peptides disclosed herein, or peptides having the formula disclosed herein, can be prepared using any of a number of well-known procedures utilizing a variety of resins and reagents.

[0262] Solid-phase peptide synthesis is a method well-known and practiced in the art. In such a method, the peptide of the present invention can be synthesized by sequentially incorporating the desired amino acid residues one at a time into the growing peptide chain, according to the general principles of solid-phase methods.

[0263] In the chemical synthesis of peptides, the reactive side chain groups of various amino acid residues are protected by suitable protecting groups to prevent a chemical reaction from occurring at that site until the protecting group is removed. Similarly, it is common practice to protect the α-amino group of an amino acid residue or fragment when the entity reacts at a carboxyl group, and then selectively remove the α-amino protecting group to allow subsequent reactions at that site. Specific protecting groups have been disclosed and are known in both solid-phase and liquid-phase synthetic methods.

[0264] The α-amino group can be protected by suitable protecting groups, including urethane-type protecting groups (such as benzyloxycarbonyl (Z) and substituted benzyloxycarbonyl groups, such as p-chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p-biphenyl-isopropoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc) and p-methoxybenzyloxycarbonyl (Moz)) and aliphatic urethane-type protecting groups (such as tert-butoxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropoxycarbonyl, and allyloxycarbonyl (Alloc)). Fmoc is particularly suitable for protecting α-amino groups.

[0265] The guanidine group can be protected by suitable protecting groups such as nitro, p-toluenesulfonyl (Tos), Z, pentamethylsuccinylsulfonyl (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 triphenylmethyl (Trt) for His.

[0266] The linear peptide precursors of the peptides of the present invention described herein are prepared using solid-phase synthesis with an automated peptide synthesizer, using the programming module provided by the manufacturer, and following the protocol outlined in the manufacturer's manual.

[0267] The linker from the side-chain amino group to the C-terminal carboxyl group of the straight-chain peptide is a straight-chain alkyl amino acid, which has no side chain except for hydrogen. Therefore, the linker can be glycine, β-alanine, γ-aminobutyric acid, 5-aminovaleric acid, 6-aminohexanoic acid, 7-aminoheptanoic acid, 8-aminooctanoic acid, etc.

[0268] On one hand, solid-phase synthesis is employed, starting from the C-terminus of the peptide, where a linear alkyl amino acid is coupled to a suitable resin to form a starting resin, and then a protected α-amino acid is coupled to the linear alkyl amino acid. For example, a pre-loaded triphenylmethyl group can be used, which is attached to an Fmoc-protected linear alkyl amino acid, such as:

[0269] Fmoc-8-aminooctanoic acid (ChemImpex, catalog number 04945)

[0270] Fmoc-7-aminoheptanoic acid (ChemImpex, catalog number 07157)

[0271] Fmoc-6-aminohexanoic acid (ChemImpex, catalog number 02490)

[0272] Fmoc-5-aminovaleric acid (ChemImpex, catalog number 04797)

[0273] Fmoc-γ-aminobutyric acid (ChemImpex, catalog number 02692)

[0274] Fmoc-β-alanine (ChemImpex, catalog number 02374)

[0275] Fmoc-glycine (ChemImpex, catalog number 02416)

[0276] However, other resins, such as Merrifield resin, Wang resin, benzyl bromoresin, 2-chloro-triphenylmethyl resin, or other resins used for the production of peptide acids, can be used. Generally, the cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, can be readily synthesized using known conventional procedures for forming peptide bonds between amino acids. Such conventional procedures include, for example, any solution-phase procedure that allows condensation between a free α-amino group of an amino acid residue (having its protected carboxyl group and other reactive groups) and a free primary carboxyl group of another amino acid residue (having its protected amino group or other reactive groups). The peptides disclosed herein, or peptides having the formula disclosed herein, can be prepared using any of a number of well-known procedures utilizing a variety of resins and reagents.

[0277] For peptide 16 of this invention, synthesis began with manual preloading of 2-chlorotriphenylmethyl chloride resin (ChemImpex, catalog 03498, 0.9 g, 1.0 mmol) containing Fmoc-5-aminovaleric acid (Fmoc-5-Ava-OH, ChemImpex, catalog 04797, 1.4 g, 4.0 mmol). The resulting Fmoc-5-Ava-2Cl triphenylmethyl resin (approximately 1.0 mmol) was loaded onto a peptide synthesizer. Then, the Fmoc-protected amino acids Trp (Boc), Arg (Pbf), D-Phe (4-F), His (Trt), Dab (Boc), and Nle were sequentially coupled individually.

[0278] After Fmoc is deprotected from Fmoc-Nle, the resulting N-terminal amino group is acylated (e.g., by using acetic anhydride and pyridine in DMF) for an appropriate time period to produce a peptide-resin:

[0279] Ac-Nle-Dab(Boc)-His(Trt)-D-Phe(4-F)-Arg(Pbf)-Trp(Boc)-NH(CH2)4COO-resin

[0280] The peptide-resin was mixed with 30 mL of a cleavage solution containing TFA / TIS / H2O (95:2.5:2.5, v / v / v) for 20 minutes, which also cleaved the orthogonal protecting groups. This was then repeated with another 30 mL of freshly prepared cleavage solution for 20 minutes. The combined filtrate was stored at room temperature for two hours and then concentrated using a purge of N2. The cleaved linear peptide was precipitated from cold diethyl ether. The solid / oily residue was dissolved in 50% tert-butanol / water and lyophilized to produce a crude linear peptide (approximately 1.0 mmol).

[0281] Ac-Nle-Dab-His-D-Phe(4-F)-Arg-Trp-NH-(CH2)4-COOH

[0282] The crude linear peptide (approximately 1.0 mmol) was dissolved in a mixture of 7.5 mL DMF and 7.5 mL DCM and cooled in an ice-cold water bath. EDC (0.288 g, 1.5 mmol) and HOAt (0.75 mmol in 0.45 mL DMF) were added to the cooled solution, followed by triethylamine (TEA) (0.4 mL, 3.0 mmol) to pH 8–9. The reaction mixture was stirred while being warmed to room temperature and left overnight at room temperature. LC / MS analysis showed that cyclization was complete. 5 mL of 1N HCl was added to the reaction mixture to adjust the pH to 3–4, and the mixture was stirred for 1 hour. The reaction mixture was filtered through a 0.45 μm syringe filter and loaded directly onto a preparative HPLC. The purified fractions were combined and lyophilized to 73 mg of cyclic peptide (6% yield).

[0283]

[0284] Other peptides of the present invention can be prepared by similar means.

[0285] Typically, each deprotection step may include, for example, the use of piperidine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), 1-hydroxybenzotriazole (HOBT), N,N-dimethylformamide (DMF), etc., followed by a washing cycle such as with DMF or methyl tert-butyl ether (MBTE), wherein the cycle is repeated as appropriate.

[0286] Each coupling step may include, for example, the use of a desired protected amino acid, such as FMOC-AA-OH, wherein the coupling agent includes dichloromethane (DCM), HOBT, N,N-diisopropylethylamine (DIPEA), DMF, or 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea hexafluorophosphate (HBTU), etc. Subsequent coupling wash cycles, such as with DMF or MBTE, may be used.

[0287] When a synthesized peptide is coupled to a resin or in solution, the N-terminus can be modified, such as by acetylation. In one approach, the resin-bound peptide is reacted with acetic anhydride in dichloromethane in the presence of an organic base (such as diisopropylethylamine) after the N-terminal protecting group has been removed. Other methods of N-terminal acetylation are known in the art, including solution-phase acetylation, and can be used.

[0288] The resin-bound peptides obtained from the resin can be cleaved by any means known in the art, such as mixing the resin-bound peptides with a mixture of trifluoroacetic acid (TFA), triisopropylsilane (TIS), and water (e.g., TFA / TIS / H2O, 95:2.5:2.5, v / v / v) for a suitable time period, such as 20 minutes, at a suitable temperature (e.g., room temperature). If necessary, one or more cycles of mixing the resin-bound peptides with the TFA / TIS / H2O mixture can be performed after filtration. The combined filtrate can be stored at room temperature for a period of time, such as two hours, and then concentrated by purging with a stream of N2. The cleaved linear peptides can then be precipitated from cold ether, and the resulting residue can be dissolved in 50% tert-butanol / water and lyophilized to produce linear peptides.

[0289] The crude linear peptide can then be cyclized in solution using conventional reaction methods for cyclization via amide bond condensation. First, the linear peptide is dissolved in a suitable solvent, such as DMF, tetrahydrofuran (THF), DCM, or 1-methyl-2-pyrrolidone (NMP). Suitable cyclic coupling agents include, for example, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TBTU), HBTU, benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate (BOP), benzotriazol-1-yl-oxy-tris(pyrrolidine)phosphonium hexafluorophosphate (PyBOP), 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TATU), 2-(2-oxo-1(2H)-pyridyl)-1,1,3,3-tetramethylurea tetrafluoroborate (TPTU), or N,N'-dicyclohexylcarbodiimide / 1-hydroxybenzotriazole (DCCI / HOBt). Conventionally, coupling is initiated by using a suitable base (e.g., DIPEA, sym-trimethylpyridine, or N-methylmorpholine (NMM)).

[0290] After cyclization of the solution, the resulting mixture can be concentrated by known means, and then it can be partially purified, such as by grinding with methyl tert-butyl ether (MTBE). The resulting fractions can then be combined and lyophilized.

[0291] Typically, orthogonal protecting groups can be used appropriately. For example, the peptides of the present invention contain a variety of amino acids having side chains containing amino groups. Any of a variety of protecting groups can be used, including the allyl-alloc protection scheme having certain amino acids, and orthogonal protecting groups that are cleavable under different reaction conditions for other amino acids having side chains containing amino groups. Thus, for example, amino acids having side chains containing amino groups can 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 similarly; by way of example and not limitation, Mtt (4-methyltriphenylmethyl) or Mtt / OPp (4-methyltriphenylmethyl / 2-phenylisopropyl) can be used with the side chain of His, wherein the orthogonal protecting group is used for other non-cleavable positions using conditions suitable for cleavage of Mtt or Mtt / OPp.

[0292] Reactive groups in peptides can be selectively modified during solid-phase synthesis or after removal from the resin. For example, when on a resin, a peptide can be modified to obtain N-terminal modifications (such as acetylation), or it can be modified after removal from the resin using a cleavage agent. Similarly, methods for modifying amino acid side chains are well known to those skilled in the art of peptide synthesis. The choice of modification to the reactive groups present on the peptide will depend in part on the desired characteristics of the peptide.

[0293] Although the synthesis has been described primarily with reference to solid-phase Fmoc chemistry, it should be understood that other chemical and synthetic methods may be used to prepare the cyclic peptides of the present invention, such as Boc chemistry, solution chemistry, and other chemical and synthetic methods, by way of example and not limitation.

[0294] 6.0 Preparations.

[0295] Depending on the desired route of administration, formulations comprising one or more cyclic peptides or cyclic peptides having the formula disclosed herein can be modified. Therefore, such formulations 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 disorders), buccal or other mucosal application, other transdermal applications, etc. Generally, formulations can be used for any form of administration of the peptides of the present invention.

[0296] 6.1 Salt forms of cyclic peptides.

[0297] The cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, can be in the form of any pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic alkali or acid (including inorganic or organic bases and inorganic or organic acids). Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese salts, divalent manganese salts, potassium salts, sodium salts, zinc salts, etc. Particularly preferred are ammonium salts, calcium salts, lithium salts, magnesium salts, potassium salts, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic alkaloids include the following: 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-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

[0298] When the cyclic peptide disclosed herein, or a cyclic peptide having the formula disclosed herein, is basic, the acid addition salt can be prepared from a pharmaceutically acceptable, non-toxic acid (including inorganic and organic acids). Such acids include 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, hydroxyethylsulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, malonic acid, mucoic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, propionic acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, TFA, etc. The acid addition salt of the peptide disclosed herein, or a peptide having the formula disclosed herein, is prepared from the peptide and an excess of acid in a suitable solvent, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, TFA, citric acid, tartaric acid, maleic acid, succinic acid, or methanesulfonic acid.

[0299] Acetate, ammonium acetate, and TFA salt forms are particularly suitable. When the peptide disclosed herein or a peptide having the formula disclosed herein includes an acidic moiety, a suitable pharmaceutically acceptable salt may include an alkali metal salt, such as a sodium or potassium salt, or an alkaline earth metal salt, such as a calcium or magnesium salt. It should also be understood that certain peptides having formulas (I) to (V) may exist in both solvated (including solvates of free peptides or solvates of salts of compounds) and non-solvated forms. The term "solvate" is used herein to describe molecular complexes comprising compounds of the invention and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol). When the solvent is water, the term "hydrate" is used. It should be understood that all polymorphs (including mixtures of different polymorphs) are included within the scope of the claimed peptide.

[0300] 6.2 Pharmaceutical composition.

[0301] This invention provides a pharmaceutical composition comprising a cyclic peptide disclosed herein or a cyclic peptide having a formula disclosed herein, and a pharmaceutically acceptable carrier. The carrier may be a liquid formulation, and preferably a buffered isotonic aqueous solution. The pharmaceutically acceptable carrier further includes excipients (such as diluents, carriers, etc.) and additives (such as stabilizers, preservatives, solubilizers, buffers, etc.), as described below.

[0302] The cyclic peptide compositions disclosed herein, or cyclic peptide compositions having the formula disclosed herein, can be formulated or compounded into pharmaceutical compositions comprising at least one cyclic peptide disclosed herein, or a cyclic peptide having the formula disclosed herein, together with one or more pharmaceutically acceptable carriers, including excipients (such as diluents, carriers, etc.) and additives (such as stabilizers, preservatives, solubilizers, buffers, etc.) (as needed). Excipients for formulations may include polyvinylpyrrolidone, gelatin, hydroxypropyl cellulose, gum arabic, polyethylene glycol, mannitol, sodium chloride, and sodium citrate. For injectable or other liquid application formulations, water containing at least one or more buffering components is preferred, and stabilizers, preservatives, and solubilizers may also be used. For solid application formulations, any of a variety of thickeners, fillers, swelling agents, and carrier additives, such as starch, sugars, cellulose derivatives, fatty acids, etc., may be used. For topical application formulations, any of a variety of creams, ointments, gels, lotions, etc., may be used. For most pharmaceutical formulations, the inactive ingredient will constitute the majority of the formulation by weight or volume. For pharmaceutical formulations, it is also considered that any of a variety of measurement-release, sustained-release, or continuous-release formulations and additives may be used, such that a dose may be formulated to deliver the peptide disclosed herein or a peptide having the formula disclosed herein over a period of time.

[0303] Typically, the actual amount of the cyclic peptide disclosed herein or having the formula disclosed herein administered to a patient will vary within a fairly wide range, depending on the method of administration, the formulation used, and the desired response.

[0304] In practical applications, the cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, can be combined with a drug carrier as active ingredients to form mixtures using conventional drug compounding techniques. The carrier can take various forms, depending on the desired formulation for administration, such as oral, parenteral (including intravenous), urethral, ​​vaginal, nasal, buccal, sublingual, etc. When preparing compositions for oral dosage forms, any common drug medium can be used, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, etc., in the case of oral liquid formulations (e.g., suspensions, elixirs, and solutions); or carriers such as starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc., in the case of oral solid formulations (e.g., powders, hard capsules, soft capsules, and tablets).

[0305] Tablets and capsules represent advantageous forms of oral dosage units due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques if desired. The amount of active peptide in such therapeutically useful compositions should be such that an effective dose can be obtained. Another advantageous form of dosage unit is the use of sublingual formulations, such as thin sheets, wafers, tablets, etc.

[0306] Tablets, pills, capsules, etc., may also contain binders such as povidone, tragacanth gum, gum arabic, 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; colorants; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose, or saccharin. When the dosage unit is in capsule form, it may also contain a liquid carrier such as fatty oil, in addition to the above types of materials. Various other materials can be used as coatings or to modify the physical form of the dosage unit. For example, tablets can be coated with shellac, sugar, or both. In addition to the active ingredient, syrups or elixirs may also contain sucrose as a sweetener, methylparaben and propylparaben as preservatives, dyes, and flavorings (such as cherry or orange flavoring).

[0307] If formulated for oral delivery, the peptide can be formulated and prepared such that it is encapsulated in an enteric protective agent, more preferably such that it is not released until the tablet or capsule passes through the stomach and optionally further through a portion of the small intestine. In the context of this application, the term enteric coating or material should be understood to refer to a coating or material that will pass substantially intact through the stomach but will disintegrate after passing through the stomach to release the active pharmaceutical ingredient. Materials that can be used include cellulose acetate phthalate, hydroxypropyl methyl ethyl cellulose succinate, hydroxypropyl methyl cellulose phthalate, polyvinyl acetate phthalate, and methyl methacrylate copolymer. The enteric coating used primarily promotes the dissolution of the dosage form at sites outside the stomach and is optionally made to dissolve at a pH of at least about 5.5, more preferably at a pH of about 6.0 to about 8.0.

[0308] Any of a variety of permeability enhancers can be used to increase intestinal uptake upon dissolution of the enteric coating. In one aspect, permeability enhancers increase extracellular or transcellular transport systems. Representative, non-limiting examples of such permeability enhancers include calcium chelators, bile salts (such as sodium cholate), and fatty acids. In some embodiments, peptides or polypeptides serving as substrates for intestinal proteases are further added.

[0309] Cyclic peptides can also be administered parenterally. Solutions or suspensions of these active peptides can be prepared in water appropriately mixed with a surfactant, such as hydroxypropyl cellulose. Dispersions can also be prepared in oils with glycerol, liquid polyethylene glycol, or mixtures thereof. These formulations may optionally contain preservatives to prevent microbial growth.

[0310] Suitable injectable drug forms include sterile aqueous solutions or dispersions, as well as sterile powders for the ad hoc preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must have sufficient flowability to be administered via a syringe. The form must be stable under the conditions of manufacture and storage and must be preserved against contamination by microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, or liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

[0311] The cyclic peptides disclosed herein can be used for therapeutic application via nasal 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 permeation enhancers, transcellular permeation enhancers, mucosal adhesion polymers, and various carrier systems. The peptides can also be formulated as dry or powder formulations. The cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, can be formulated with any of a variety of pharmaceutical agents that enhance the effective nasal absorption of drugs, including peptide drugs. These pharmaceutical agents can enhance nasal absorption without causing unacceptable damage to the mucosa. U.S. Patent Nos. 5,693,608, 5,977,070, and 5,908,825, etc., teach numerous pharmaceutical compositions (including absorption enhancers) that can be used, and the teachings of each of these, along with all references and patents cited therein, are incorporated herein by reference.

[0312] In aqueous solutions, cyclic peptides can be appropriately buffered by saline, acetate, phosphate, citrate, or other buffers, which can have any physiologically acceptable pH, typically from about pH 4 to about pH 7. Combinations of buffers, such as phosphate-buffered saline, saline, and acetate buffers, can also be used. In the case of saline, a 0.9% saline solution can be used. In the case of acetate, phosphate, citrate, etc., a 50 mM solution can be used. In addition to buffers, suitable preservatives can be used to prevent or limit the growth of bacteria and other microorganisms. One such preservative that can be used is 0.05% benzalkonium chloride.

[0313] In alternative embodiments, the cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, can be administered directly to the lungs. Intrapulmonary administration, when driven by a patient during inhalation, can be performed via a metered-dose inhaler (a device that allows self-administration of a dose-dispensed peptide or peptide having the formula disclosed herein). In one aspect of this embodiment, the cyclic peptides can be in the form of dry particles, for example, particles between about 0.5 and 6.0 μm, such that the particles have sufficient mass to settle on the lung surface without being exhaled, but are small enough that they do not deposit on the airway surface before reaching the lungs. Dry powder microparticles can be prepared using any of a variety of different techniques, including but not limited to micromilling, spray drying, and quick-freezing aerosol followed by lyophilization. In the case of microparticles, the peptides can deposit deep into the lungs, thereby being rapidly and efficiently absorbed into the bloodstream. Furthermore, such methods do not require penetration enhancers, as is sometimes the case in transdermal, nasal, or oral mucosal delivery routes. Any of a variety of inhalers can be used, including propellant-based nebulizers, aerosols, single-dose dry powder inhalers, and multi-dose dry powder inhalers. Commonly used devices include metered-dose inhalers, which are used to deliver medications for the treatment of asthma, chronic obstructive pulmonary disease, etc. Preferred devices include dry powder inhalers, which are designed to form a fine powder cloud or aerosol with a particle size consistently less than about 6.0 μm.

[0314] Particle size (including average size distribution) can be controlled through preparation methods. For micro-grinding, particle size is controlled by factors such as the size of the grinding head, rotor speed, and processing time. For spray drying, particle size is controlled by factors such as nozzle size, flow rate, and dryer temperature. For preparations via quick-freezing aerosols followed by freeze-drying, particle size is controlled by factors such as nozzle size, flow rate, and aerosol solution concentration. These and other parameters can be used to control particle size.

[0315] The cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, can be therapeutically administered via an injectable sustained-release formulation. In one embodiment, the cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, are formulated into a formulation for deep intramuscular injection (e.g., in the gluteal or deltoid muscles), the formulation comprising 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, pH-adjusting sodium hydroxide or hydrochloric acid, etc. In another embodiment, the cyclic peptides disclosed herein, or cyclic peptides having the formula disclosed herein, are formulated with poly(orthoester) (which may be an autocatalytic poly(orthoester) having any variable percentage of lactic acid in the polymer backbone) and optionally one or more additional excipients. In one embodiment, a poly(D,L-lactide-co-glycolic acid) polymer is used. Generally, any of many injectable and biodegradable polymers (preferably also viscous polymers) can be used in sustained-release injectable formulations. Alternatively, other sustained-release formulations may be used, including those permitted for subcutaneous injection. These other formulations may include one or more of nanospheres / microspheres (such as compositions comprising PLGA polymers), liposomes, emulsions (such as water-in-oil emulsions), gels, suspensions in insoluble salts, or oils. Depending on the concentration and amount of the cyclic peptide, the sustained-release rate of the materials used, and other factors known to those skilled in the art, the formulation may require injections on a daily, weekly, monthly, or other periodic basis.

[0316] 6.3 Application route.

[0317] If a composition comprising one or more peptides disclosed herein or having the formula disclosed herein is administered by injection, the injection may be intravenous, subcutaneous, intramuscular, intraperitoneal, or by other means known in the art. The peptides disclosed herein or having the formula disclosed herein may be formulated in any manner known in the art, including but not limited to tablets, capsules, pouches, suspensions, powders, lyophilized formulations, suppositories, eye drops, skin patches, oral soluble formulations, sprays, aerosols, etc., and may be mixed and formulated with buffers, binders, excipients, stabilizers, antioxidants, and other agents known in the art. Generally, any route of administration can be used to introduce the peptides of the invention through the epidermal layer of cells. Therefore, administration methods may include mucosal administration, buccal administration, oral administration, skin administration, inhalation administration, nasal administration, urethral administration, vaginal administration, etc.

[0318] 6.4 Effective therapeutic dose.

[0319] Typically, the actual amount of the cyclic peptides disclosed herein or having the formula disclosed herein administered to a patient will vary within a fairly wide range, depending on the route of administration, the formulation used, and the desired response. A therapeutic dose is an amount sufficient to produce the desired therapeutic effect, administered by any of the foregoing means or any other means known in the art. The cyclic peptides disclosed herein or having the formula disclosed herein are generally highly active. For example, cyclic peptides may 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.

[0320] 7.0 is used for peptide assessment testing and determination.

[0321] The melanocyte-stimulating receptor-specific peptides disclosed herein, or melanocyte-stimulating receptor-specific peptides having the formula disclosed herein, can be tested using a variety of assay systems and animal models to determine their binding, functional status, and efficacy.

[0322] 7.1 Agonist activity assay performed by CEREP.

[0323] The agonist activity of compounds on the melanocyte-stimulating receptor was determined by measuring the effect of compounds on cAMP production using an HTRF assay at CEREP (CEREP SA, Eurofins, Celle-Lévescault, France). Cells were suspended in HBSS buffer (Invitrogen) supplemented with 20 mM HEPES (pH 7.4) and 500 μM IBMX, then distributed in microplates and incubated in the presence of HBSS (basal control), the test compound, or a reference agonist. Incubation time, temperature, cell number, reference agonist, and cell line information are included in the table below. For the stimulated control assay, a separate assay well contained the reference compound. After incubation, cells were lysed and the fluorescent receptor (D2-labeled cAMP) and fluorescent donor (anti-cAMP antibody labeled with a europium cavitation compound) were added. After 60 minutes at room temperature, fluorescence transfer at excitation wavelengths of 337 nm and emission wavelengths of 620 and 665 nm was measured using an Envision microplate reader (PerkinElmer). The cAMP concentration was determined by dividing the signal measured at 665 nm by the signal measured at 620 nm (ratio).

[0324] Results are expressed as the percentage of control response to a 1 μM reference. In each experiment, the standard reference agonist was tested at several concentrations to generate concentration-response curves from which EC50 was calculated. 50 value.

[0325]

[0326] 7.2 Antagonist activity assay performed by CEREP.

[0327] This assay was used to evaluate the antagonistic activity of the compound against the melanocyte-stimulating receptor, which was determined by measuring its effect on cAMP production using an HTRF assay.

[0328] Cells with desired melanocyte-stimulating receptors were suspended in HBSS buffer (Ingenieur) supplemented with 20 mM HEPES (pH 7.4) and 500 μM IBMX, and then distributed in microplates containing HBSS (basal control), the test compound, or a reference antagonist. Subsequently, a concentration of agonist was added to stimulate cAMP production. For basal control measurements, individual assay wells did not contain the reference agonist. Incubation time, temperature, cell number, reference agonist, and cell line information are included in the table below.

[0329] After incubation, cells were lysed and a fluorescent receptor (D2-labeled cAMP) and a fluorescent donor (anti-cAMP antibody labeled with a europium cavitation compound) were added. After 60 minutes at room temperature, fluorescence transfer was measured using a microplate reader (Envision, PerkinElmer) at excitation wavelengths of 337 nm and emission wavelengths of 620 and 665 nm. The cAMP concentration was determined by dividing the signal measured at 665 nm by the signal measured at 620 nm (ratio). The results were expressed as the percentage of inhibition of the control response against a reference agonist. A standard reference antagonist was tested at several concentrations in each experiment to generate concentration-response curves from which the IC50 was calculated. 50 value.

[0330]

[0331] 7.3 Alternative assays for agonist activity.

[0332] Intracellular cAMP accumulation was examined as a measure of the ability of peptides to elicit functional responses in 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 the culture plate by incubation in enzyme-free cell dissociation buffer. The 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-methylxanthine (IBMX) (a phosphodiesterase inhibitor). Cells were cultured at 0.5 x 10⁻⁶ cells per well. 5Cells were randomly assigned to 96-well plates and pre-incubated for 10 minutes. Cells were then exposed to a peptide dissolved in DMSO (final DMSO concentration 1%) for 15 minutes at 37°C, with a concentration range of 0.05–5000 nM, for a total assay volume of 200 μL. NDP-α-MSH was used as a reference agonist. Assays were performed using Cisbio Bioassays. cAMP levels were determined using a cAMP-based assay system that employed crypt-labeled anti-cAMP and D2-labeled cAMP, with plate readings at 665 and 620 nM on a PerkinElmer Victor plate reader. The readings were performed using a Graph-Pad. The software uses nonlinear regression analysis to analyze the data. The maximum efficacy of the tested peptide is compared with the maximum efficacy achieved by the reference melanocyte-stimulating hormone agonist NDP-α-MSH.

[0333] 7.4 Functional determination of high-density and low-density hMC4r.

[0334] HEK293 cell line transfected with human MC4r (from Palatin Technologies, USA, licensed from the University of Michigan) was used. T-REx from Ingenium was used. TM The system introduces human MC4r into HEK293. T-REx TM The system employs a tetracycline-regulated mammalian expression system, utilizing a regulatory element from *E. coli* Tn10 encoding the tetracycline (Tet) resistance operon. This is achieved through the use of T-REx... TM The expression of the target gene (human MC4r gene) in the system was suppressed in the absence of tetracycline or doxycycline, but induced in the presence of tetracycline or doxycycline (see T-REx). TM System manual (published by Ingenium Corporation).

[0335] HEK293-T-REx-MC4r cells were cultured in DMEM (Gibco 11965) supplemented with L-glutamine (Gibco 25030), 10% fetal bovine serum (FBS), 200 μg / mL bleomycin (Ingenieur 46-0072), and 6 mg / mL blastomycin (Ingenieur 46-1120) at 5% CO2 and 95% humidity at 37°C. T-150 cell culture flasks with 75% confluence were incubated with two concentrations of doxycycline (0.1 ng / mL to provide a low-density hMC4r system and 10 ng / mL to provide a high-density hMC4r system) at 5% CO2 and 37°C for 16–18 hours to induce MC4r expression. On the day of assay, cells were washed with PBS (Gibco 14190) and harvested using cell dissociation buffer (Gibco 13150-016), then centrifuged and resuspended in Hanks balanced salt solution (+Ca,+Mg) (Gibco 14025), 10 mM 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic 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-isobutyl-1-methylxanthine (IBMX).

[0336] Then the cells were inoculated at 198 μL (approximately 5 x 10⁻� 4 Cells were randomly assigned to 96-well plates (BD 353916) and incubated at 37°C for 10 min. Cells were then exposed to a peptide dissolved in DMSO (final DMSO concentration 1%) for 15 min at 37°C, with a concentration range of 10 μg / well. -5 Up to 10 -13 M, with a total assay volume of 200 μL, in which NDP-α-MSH was used as a reference agonist. The reaction was terminated by adding 15 μL of lysis buffer to each well and shaking the plate at room temperature for 30 minutes.

[0337] Through Xisi Biometrics Company cAMP levels were determined using a cAMP-based assay system that employed crypt-labeled anti-cAMP and d2-labeled cAMP, with plate readings performed at 665 and 620 nM on a PerkinElmer Victor plate reader. The results were obtained using a Graph-Pad. The software uses nonlinear regression analysis to analyze the data. The maximum efficacy of the tested peptide is compared with the maximum efficacy achieved by the reference melanocyte-stimulating hormone agonist NDP-α-MSH.

[0338] 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, MC4r agonist stimulation increases cAMP levels. cAMP levels were measured using the cAMP Dynamics 2HTRF Kit (CisBio catalog number 62AM4PEC; see CisBio brochure). cAMP levels were normalized against plate controls (0% for 1% DMSO, 100% for 400 nM NDP-α-MSH) and calibration curves (as described in the CisBio HTRF Kit) for the range of 712 nM to 0.04 nM cAMP. Plates were incubated on a shaker for 1 hour at room temperature and read at 665 and 620 nm using a PerkinElmer Victor plate reader. Then, the fluorescence ratio was calculated as described in the Xisi Biotechnology HTRF kit, where GraphPad Prism software was used to plot the percentage of fluorescence relative to the calculated cAMP concentration using a variable slope dose-response curve to determine the EC50. 50 and E max value.

[0339] 8.0 Peptide structure example.

[0340] In one aspect, a cyclic peptide is provided, which contains a core sequence derived from the His-Phe-Arg-Trp sequence or a modification thereof in its cyclic moiety. This peptide is cyclized via a side chain of an adjacent amino acid (on the N-terminal side), His (or a derivative, modification, or substitution of His), and a C-terminal group. The cyclic peptide is at least a cyclic pentapeptide (containing five amino acids within the cyclic moiety) and optionally a cyclic hexapeptide, heptapeptide, or larger cyclic peptide, wherein one or more additional amino acid residues are present outside the N-terminal cyclic moiety.

[0341] For MC4r antagonists that may simultaneously contain MC1r, MC3r, or MC54 agonists or combinations thereof, in some respects, the core sequence derived from His-Phe-Arg-Trp will include D-Phe at the Phe position instead of L-Phe, with Nal 1 or Nal 2 substitution (at the Phe position) (e.g., D-Nal 1 or D-Nal 2), or alternatively may include substituted Phe at the Phe position, such as substituted D-Phe or substituted L-Phe. Various amino acids may be used for the remaining amino acids in the core sequence. Typically, the His position may be substituted or unsubstituted Pro, or may be an amino acid with a side chain comprising at least one primary amine, secondary amine, alkyl, cycloalkyl, heteroalkyl, aryl, heteroaryl, alcohol, ether, sulfide, sulfone, sulfoxide, carbamoyl, or carboxyl group. The Arg position can be a substituted or unsubstituted Pro, or it can be an amino acid with a side chain comprising at least one primary amine, secondary amine, guanidine, urea, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, or ether. The Trp position can be an amino acid with a side chain comprising at least one substituted or unsubstituted aryl or heteroaryl group, or alternatively, it can be omitted.

[0342] The peptides covered by formulas (I), (II), (III), (IV), and (V) contain one or more asymmetric elements, such as stereoisomer centers, stereoisomer axes, etc., and therefore such peptides covered by these formulas can exist in different stereoisomeric forms. For the peptides specifically and generally described, including those covered by formulas (I), (II), (III), (IV), and (V), all isomeric forms (including enantiomers and diastereomers) of all chiral or other isomeric centers are covered herein. The peptides of the present invention each comprise multiple chiral centers and, in addition to being used in enantiomerically pure formulations, can also be used as racemic mixtures or mixtures enriched with enantiomers. Typically, the peptides of the present invention are synthesized using chiral pure reagents (such as specified L- or D-isomer amino acids), using reagents, conditions, and methods that maintain enantiomeric purity, but racemic mixtures can be prepared, and are contemplated. Such racemic mixtures can optionally be separated using well-known techniques, and individual enantiomers can be used alone. In the context of the temperature, solvent, and pH conditions under which peptides can exist in tautomeric forms, each tautomeric form is considered to be included in this invention, whether present in equilibrium or primarily in one form. Therefore, a single enantiomer (optionally in its active form) of a peptide having formulas (I) to (V) can be obtained by asymmetric synthesis, synthesis from an optically pure precursor, or resolution of a racemic mixture.

[0343] The peptides disclosed herein are specific stereoisomers of peptides having formulas (I) to (V), but the present invention should not be construed as being limited to the stereoisomers covered by the peptides disclosed herein.

[0344] This invention further aims to include prodrugs comprising the peptides of the invention, which, upon administration, undergo chemical transformation via metabolic processes to become active pharmacological peptides. Typically, such prodrugs will be functional derivatives of the peptides of the invention, readily convertible in vivo into peptides having formulas (I) to (V). A prodrug is any covalently bonded compound that releases an active parent peptide drug having formulas (I) to (V) in vivo. Conventional procedures for selecting and preparing suitable prodrug derivatives are described, for example, in "Design of Prodrugs," edited by H. Bundgaard, Elsevier, 1985. Typical examples of prodrugs have biologically unstable protecting groups on their functional moieties, such as through esterification of hydroxyl, carboxyl, or amino functional groups. Therefore, by way of example and not limitation, prodrugs include peptides having formula (I), (II), or (III) wherein an ester prodrug form is used, such as lower alkyl esters (e.g., where R is -OH, and the alkyl group of these lower alkyl esters may include 1-8 carbons) or aralkyl esters (having 6-12 carbons in the aralkyl group) of formula (I), (II), or (III). More broadly, prodrugs include compounds that can be oxidized, reduced, amination, deamination, hydroxylation, dehydroxylation, hydrolysis, dehydration, alkylation, dealkylation, acylation, deacylation, phosphorylation, or dephosphorylation to produce in vivo an active parent peptide drug having formula (I).

[0345] This invention also includes the same peptides as those described in formula (I), but in fact, one or more atoms described in formula (I) are replaced by atoms with atomic masses or mass numbers different from those commonly found in nature. Examples of isotopes that can be incorporated into the peptides of this invention include isotopes of hydrogen, carbon, nitrogen, and oxygen, respectively as follows: 2 H, 3 H, 13 C 14 C 15 N、 18 O and 17 O. The peptides disclosed herein, or peptides having the formula disclosed herein, as well as pharmaceutically acceptable salts or solvates of said peptides (the peptide containing the aforementioned isotopes and / or other isotopes of other atoms) are within the scope of this invention. Certain isotope-labeled peptides, such as radioactive isotopes (e.g., 3 H and 14 C) The incorporated peptides can be used in various assays, such as in drug and / or substrate tissue distribution assays. In some cases, heavier isotopes are substituted, such as deuterium (…). 2Replacing one or more hydrogen atoms with H can provide pharmacological advantages, including improved metabolic stability. Isotopically labeled peptides of formula (I) can often be prepared by replacing non-isotopically labeled reagents with isotopically labeled reagents.

[0346] 9.0 instance.

[0347] The present invention is further illustrated by the following non-limiting examples:

[0348] Peptides with the following structures were synthesized using the general methods described above, and the EC values ​​of the peptides were determined as shown. 50 Value. ECs marked with "*" 50 The value is determined by CEREP. In EC 50 In the case of values, "%" indicates the percentage of Emax (the percentage of the maximum response obtained with a positive control). "NC" indicates the EC50 value. 50 The value represents EC 50 The value exceeds 10,000 nM and therefore cannot be calculated.

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357] While the 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 invention will be apparent to those skilled in the art, and are intended to cover all such modifications and equivalents. All references, applications, patents, and the full disclosure thereof cited above are hereby incorporated by reference.

Claims

1. A cyclic peptide selected from one of the group consisting of the following compounds: ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， , and 。

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