Peptide compositions

Abstract

To provide polypeptide compounds that are modulators (e.g., agonists and antagonists) of the melanocortin-4 receptor (MC4R) and pharmaceutical compositions comprising the same.SOLUTION: The present invention provides polypeptide of the following structural Formula (I) or a pharmaceutically acceptable salt thereof. In the formula, R1 is -H or C1-C6 acyl; R2 is -NR3R4 or -OR5, and R3, R4, and R5 are each independently H or C1-C6 alkyl.SELECTED DRAWING: None

Description

Technical Field

[0001] This application claims the benefit of U.S. Provisional Application No. 61 / 790,469, filed on Mar. 15, 2013. The teachings of the above application are incorporated herein by reference.

Background Art

[0002] Disorders such as obesity, metabolic syndrome, insulin resistance and diabetes significantly increase the cost of national health management and can seriously affect the quality of life of the affected individuals, their families and caregivers. The incidence of these disorders is increasing and approaching epidemic proportions. Therefore, there is a need for compositions and methods for treating these disorders.

Summary of the Invention

[0003] The present invention relates to a polypeptide compound that is a modulator of melanocortin-4 receptor (MC4R), and a pharmaceutical composition containing the same.

[0004] In certain embodiments, the polypeptide compound is an isolated polypeptide of the following structural formula (I):

Chemical Formula

[0005] or a pharmaceutically acceptable salt thereof, wherein, R 1 is -H or C1-C6 acyl; R 2 is -NR 3 R 4 or -OR 5 and R 3 R 4 and R 5 are each independently​​​​​​​ and is H or C1-C6 alkyl; A 1 is an amino acid residue selected from Arg, Lys, Orn, His, Nle, Phe, Val, Leu, Trp , Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzAl or; A 1 is optionally substituted C1-C12 alkyl, optionally substituted C6-C 18 aryl, optionally substituted C5-C18 heteroaryl, aralkyl where the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl where the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; or; A and A are each independently an amino acid residue selected from Cys, hCys, Pen, Asp, Glu, Lys, Orn, Dbu, or Dpr, and A and A 2 are selected as a pair such that a covalent bond can be formed between the respective side chains; 8 A is absent or is an amino acid residue selected from Ala, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib or is residue Y where Y has the following structural formula 2 and A 8 is absent or is an amino acid residue selected from Ala, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib or is residue Y where Y has the following structural formula A 3 is absent or is an amino acid residue selected from Ala, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib or is residue Y where Y has the following structural formula sn, Aib or is residue Y where Y has the following structural formula structural formula

Chemical formula

[0006] an amino acid selected from the amino acids represented by R 11 and R 12 are each independently H, -CH3, phenyl or benzyl ; R 21 , R 22 , R 23 and R 24 are each independently H, -CH3, -CF3 , phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; R 31 , R 32 , R 33 , R 34 , R 41 , R 42 and R 43 are each independently , H, -CH3, -CF3, phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; A 4 is absent or is an amino acid selected from Atc, Ala, QAla, Aib, Sar, Ser , Thr, Pro, Hyp, Asn, Gln, optionally substituted His, Trp, T yr, Lys, Arg, sChp, or the residue X, where X is an amino acid selected from the amino acids represented by the following structural formula

Chemical formula

[0007] and is an amino acid selected from the amino acids represented by wherein R 51 and R 52 are each independently H, -CH3, phenyl, or benzyl ; R 61 , R 62 , R 63 and R 64are each independently H, -CH3, -CF3 , phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; R 71 R 72 R 73 R 74 R 81 R 82 and R 83 are each independently , H, -CH3, -CF3, phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; A 5 is optionally substituted Phe, optionally substituted 1-Nal, or optionally substituted 2-Nal; A 6 is Arg; A 7 is Trp, any amino acid residue is either in the L-conformation or the D-conformation, provided that (1)A 3 and A 4 are not both absent; (2)when A 4 is an amino acid, A 3 is not Aib or Gly; (3)when A 4 is His and A 5 is D-Phe or 2-Nal, A 3 is not a D-amino acid or L-Ala; (4)when A 2 and A 8 are each selected from Cys, hCys or Pen, then (a)when A 4 is absent, A 3 is not L-His; (b)when A 3 is absent, A 4 is not L-His; (c)when A 4 is His, A3 is a polypeptide that is not Glu, Leu, or Lys.

[0008] The present invention also relates to a method of treating a disorder responsive to modulation of MC4R in a subject in need thereof. The method comprises administering to the subject an effective amount of the MC4R modulation described herein. In certain embodiments, disorders responsive to modulation of MC4R include type 1 diabetes, type 2 diabetes, obesity, insulin resistance, metabolic syndrome, male erectile dysfunction, female sexual dysfunction, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, alcohol use disorder, substance use disorder including substance abuse, cachexia, inflammation, and anxiety.

[0009] In certain embodiments, the compounds and compositions of the present invention have high selectivity and efficacy for MC4R and melanocortin-3 receptor (MC3R) compared to melanocortin-1 receptor (MC1R). The compounds and compositions of the present invention can reduce or eliminate undesirable side effects such as increased blood pressure effects, increased heart rate, sexual stimulation, and increased skin pigmentation. DETAILED DESCRIPTION OF THE INVENTION

[0010] Examples of embodiments of the present invention are described as follows.

[0011] (TERMINOLOGY) The terminology used to define the peptide is typically the terminology used in the art, where the N-terminal amino group appears on the left and the C-terminal carboxyl group appears on the right.

[0012] As used herein, the term "amino acid" includes both naturally occurring amino acids and non-natural amino acids. Unless otherwise specified, all amino acids and their residues found in the compounds described herein may be in either the D-configuration or the L-configuration. Compounds of the invention useful for practicing the methods described herein may have one or more chiral centers and thus may exist in many stereoisomeric forms. All stereoisomers and mixtures thereof are included within the scope of the invention. Racemic compounds may be separated using either preparative HPLC with a chiral stationary phase and column or resolved using methods known to those skilled in the art to obtain the individual enantiomers.

[0013] In addition, chiral intermediate compounds may be resolved and used to prepare chiral compounds of the invention. The compounds described herein may exist in one or more tautomeric forms. All tautomers and mixtures thereof are included within the scope of the invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications, patent applications, patents, and other references mentioned herein are incorporated herein by reference.

[0014]

[0015] [Table 1] TIFF2025102830000005.tif192149 TIFF2025102830000006.tif158139​​​​​​​​​​

[0016] Unless otherwise specified, all abbreviations of amino acids in the present disclosure (e.g., Ala) refer to amino acid residues (i.e., represent the structure -NH-C(R)(R’)-CO-, where R and R ’ are each independently hydrogen or an amino acid side chain (e.g., in the case of Ala, R = CH3 and R’ = -H, or R and R’ may be connected to form a ring system).

[0017] The notations “Ac” or “NH2” at the polypeptide termini indicate that the corresponding termini are acylated or amidated, respectively.

[0018] The phrase “covalent bond between amino acid side chains” means that the side chains of the two amino acid residues in question each contain functional groups capable of forming a covalent bond with each other. Examples of such bonds include disulfide bridges formed by Cys side chains, hCys side chains or Pen side chains, and amide bonds formed by the amino group of one amino acid side chain and the carboxy group of another amino acid side chain (e.g., Asp, Glu, Lys, Orn, Dbu or Dpr). In an exemplary embodiment, the amino acids can be selected in pairs such that their respective side chains can form a covalent bond between them. When a covalent bond is formed between amino acid side chains, the polypeptide may cyclize. Such cyclic polypeptides may be indicated by a structural formula or by using the shorthand “c()” or “cyclo()”. For example, “-c(Cys-Cys)-” or “-cyclo (Cys-Cys)-” represents ​​​​​​​​

[0019] shows the structure, while "-c(Asp-Lys)-" or "-cyclo(Asp-Lys )-" represents the structure

Chemical formula

[0020] is shown.

[0021] "Alkyl" used alone or as part of a larger moiety such as "hydroxyalkyl", "alkoxyalkyl", "a lkylamine" refers to a straight-chain or branched-chain saturated aliphatic group having a specific number of carbons and typically having 1 to 12 carbon atoms. More specifically, the aliphatic group may have 1 to 8, 1 to 6, or 1 to 4 carbon atoms. This term is exemplified by groups such as methyl, ethyl, n-propyl, i sopropyl, n-butyl, isobutyl, tert-butyl, n-hexyl, etc.

[0022] "Haloalkyl" refers to an alkyl group substituted with one or more halogen atoms.

[0023] "Halogen" and "halo" refer to fluoro, chloro, bromo, or iodo.

[0024] "Cyano" refers to the -CN group.

[0025] "Ph" refers to the phenyl group.

[0026] "Carbonyl" refers to the divalent -C(O)- group.

[0027] Used alone or as part of a larger moiety such as "aralkyl" ​​The "aryl" used herein refers to an aromatic carbocyclic group having 6 to 18 carbon atoms and having one or more fused rings. The term "aryl" also includes (one or more) aromatic carbocyclic rings fused to a cycloalkyl group or a heterocycloalkyl group. Examples of aryl groups include phenyl, benzo[d][1,3]dioxole, naphthyl, phenanthrenyl, etc.

[0028] "Aryloxy" refers to an -OAr group, where O is an oxygen atom and Ar is an aryl group as defined above.

[0029] "Aralkyl" refers to an alkyl in which at least one alkyl hydrogen atom is replaced by an aryl moiety, such as benzyl, -(CH2)2phenyl, -(CH2)3phenyl, -CH(phenyl)2, etc.

[0030] "Heteroaryl", used alone or as part of a larger moiety such as "heteroaralkyl", refers to a monocyclic, bicyclic or tricyclic heteroaromatic ring system of 5 to 18 members containing 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur. The term "heteroaryl" also includes (one or more) heteroaromatic rings fused to a cycloalkyl group or a heterocycloalkyl group. Specific examples of heteroaryl groups include optionally substituted pyridyl, pyrrolyl, pyrimidinyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl. Diaazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl, benzofuryl, [2,3-dihydro]benzofuryl, isobenzofuryl, benzothienyl, benzotriazo ryl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzui midazolyl, imidazo[1,2-a]pyridyl, benzothiazolyl, benzoxazolyl , quinolidinyl, quinazolinyl, phthalazinyl, quinoxalinyl, cinnolinyl, naphthi ridinyl, pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4 ,3-b]pyridyl, quinolyl, isoquinolyl, tetrazolyl, 1,2,3,4-tetra hydroquinolyl, 1,2,3,4-tetrahydroisoquinolyl, purinyl, pteridinyl , carbazolyl, xanthenyl, benzquinolyl, and the like.

[0031] "Heteroalkyl" refers to an alkyl in which at least one alkyl hydrogen atom is replaced by a heteroaryl moiety, for example, -CH2-pyridinyl, -CH2-pyrimidinyl, and the like. refers to.

[0032] "Alkoxy" refers to an -O-R group, where R is "alkyl", "cycloalkyl", " alkenyl" or "alkynyl". Examples of alkoxy groups include, for example, methoxy , ethoxy, ethenoxy, and the like.

[0033] "Hydroxyalkyl" and "alkoxyalkyl" are alkyl groups substituted with hydroxy and alkoxy, respectively.

[0034] "Amino" means -NH2, and "alkylamine" and "dialkylamine" are , each representing -NHR and -NR2, where R is an alkyl group. "Cycloalkylamine" and "dicycloalkylamine" each represent -NHR and -NR2, respectively, and R is a cycloalkyl group. "Cycloalkylalkylamine" represents -NHR, and R is a cycloalkylalkyl group. "[Cycloalkylalkyl][alkyl]amine" represents -N(R)2, where R is cycloalkylalkyl and the other R is alkyl. "Acyl" refers to R”-C(O)-, where R” is H, alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl, alkenyl, substituted alkenyl, aryl, alkylaryl, or substituted alkylaryl, and is denoted as "Ac" in the general formula of certain embodiments. Substituents suitable for "alkyl", "aryl", or "heteroaryl", etc. are those that produce stable compounds of the present invention. Examples of suitable substituents are halogen, -CN, -OH, -NH2, (C1-C4)alkyl, (C1-C4)haloalkyl, aryl, heteroaryl, (C3-C7)cycloalkyl, (5-7 membered ring)heterocycloalkyl, -NH(C1-C6)alkyl, -N((C1-C6)alkyl)2, (C1-C6)alkoxy, (C1-C6)alkoxycarbonyl, -CONH2, -OCONH2, -NHCONH2, -N(C1-C6)alkylCONH2, -N(C1-C6)alkylCONH(C1-C6)alkyl, -NHCON((C1-C6)alkyl)2, -N(C1-C6)alkylCON((C1-C6)alkyl)2.

[0035]

[0036] ​​​​​​​​​​​​​​​​Le) 2, -NHC(S)NH2, -N(C1-C6)alkylC(S)NH2, -N(C 1-C6)alkylC(S)NH(C1-C6)alkyl, -NHC(S)NH(C1- C6)alkyl, -NHC(S)N((C1-C6)alkyl)2, -N(C1-C6) alkylC(S)N((C1-C6)alkyl)2, -CONH(C1-C6)alkyl , -OCONH(C1-C6)alkyl - CON((C1-C6)alkyl)2, -C( S)(C1-C6)alkyl, -S(O) p (C1-C6)alkyl, -S(O) p NH 2, -S(O) p NH(C1-C6)alkyl, -S(O) p N((C1-C6)alkyl )2, -CO(C1-C6)alkyl, -OCO(C1-C6)alkyl, -C(O) O(C1-C6)alkyl, -OC(O)O(C1-C6)alkyl, -C(O)H or -CO2H. More specifically, the substituent is selected from the group consisting of halogen, -C N, -OH, -NH2, (C1-C4)alkyl, (C1-C4)haloalkyl, (C1 -C4)alkoxy, phenyl and (C3-C7)cycloalkyl. In the backbone of the present invention, the "substitution" also means a situation where a hydrogen atom is replaced by deuterium, and p is an integer having a value of 1 or 2.

[0037] Suitable substituents for substituted Phe include 1 to 5 substituents on any aromatic carbon, and the substituents are selected from F, Cl, Br, I, -CH3, -OH, -CN, amine, - NO2, or -OCH3. Examples include Phe(2'-F), Phe( 2'-Cl), Phe(2'-Br), Phe(2'-I), Phe(2'-CN), P ​ he(2’-CH3), Phe(2’-OCH3), Phe(2’-CF3), Phe( 2’-NO2), Phe(3’-F), Phe(3’-Cl), Phe(3’-Br), Phe(3’-I), Phe(3’-CN), Phe(3’-CH3), Phe(3’- OCH3), Phe(3’-CF3), Phe(3’-NO2), Phe(4’-F), Phe(4’-Cl), Phe(4’-Br), Phe(4’-I), Phe(4’-C N), Phe(4’-CH3), Phe(4’-OCH3), Phe(4’-CF3), Phe(4’-NO2), Phe(4’-t-Bu), Phe(2’,4’-diF), P he(2’,4’-diCl), Phe(2’,4’-diBr), Phe(2’,4’-di I), Phe(2’,4’-di-CN), Phe(2’,4’-di-CH3), Phe( 2’,4’-di-OCH3), Phe(3’,4’-diF), Phe(3’,4’-diC l), Phe(3’,4’-diBr), Phe(3’,4’-diI), Phe(3’,4 ’-di-CN), Phe(3’,4’-di-CH3), Phe(3’,4’-di-OCH 3), Phe(3’,5’-diF), Phe(3’,5’-diCl), Phe(3’,5 ’-diBr), Phe(3’,5’-diI), Phe(3’,5’-di-CN), Phe (3’,5’-diCH3), Phe(3’,5’-di-OCH3), or Phe(3’ ,4’,5’-triF).

[0038] Suitable substituents for substituted His include 1 to 3 substituents on any substitutable ring atom, and the substituents are F, Cl, Br, I, -CH3, -OH, -CN, amide Selected from -NO2, benzyl, or -OCH. Examples include 1-methyl-hi Stizidine and 3-methyl-histidine.

[0039] "(Amino acid) n " means that the amino acid is repeated n times. For example, the notations "(Pro)2" or "(Arg)3" mean that the proline residue or arginine residue is repeated 2 or 3 times, respectively.

[0040] The pharmaceutically acceptable salts of the polypeptide compounds disclosed in this specification are included in the present invention. For example, the acid salts of compounds containing an amine or other basic group can be obtained by reacting this compound with a suitable organic acid or inorganic acid, and a pharmaceutically acceptable anionic salt form is obtained. Examples of anionic salts include acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylic acid salt, estolate, esilate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinolate salt, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate, pamoate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, basic acetate, succinate, sulfate, tannate, tartrate, theophyllinate, tosylate salt, etc. For example, the notations "(Pro)2" or "(Arg)3" mean that the proline residue or arginine residue is repeated 2 or 3 times, respectively. phosphate / diphosphate, polygalacturonate, salicylate, stearate, basic acetate, succinate, sulfate, tannate, tartrate, theophyllinate, tosylate salt, etc. Examples include ruthenate, triethiodide, and trifluoroacetate.

[0041] Salts of compounds containing an acid functional group can be prepared by reacting with a suitable base. Such pharmaceutically acceptable salts can be made using a base and may provide a pharmaceutically acceptable cation, including alkali metal salts (especially sodium and potassium), alkaline earth metal salts (especially calcium and magnesium), aluminum salts, and ammonium salts, as well as salts made from physiologically acceptable organic bases (e.g., trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, Ν,Ν’-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, Ν,Ν’-bisdehydroabietylamine, glucamine, N-methylglucamine, collidine, quinine, quinoline) and basic amino acids (e.g., lysine and arginine). The disclosed compounds can be administered to a subject in combination with an acceptable pharmaceutical carrier as part of a pharmaceutical composition. The formulations of the compounds to be administered will vary according to the chosen route of administration (e.g., solution, emulsion, capsule). Suitable pharmaceutical carriers may contain inert ingredients that do not interact with the compounds. Standard pharmaceutical formulation techniques may be used, for example, as described in Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, PA.

[0042] Techniques such as those may also be used. Suitable pharmaceutical carriers for topical administration include, for example, sterile water, physiological saline, bacteriostatic saline (saline containing 0.9% mg / ml benzyl alcohol ), phosphate-buffered saline, Hank's solution, Ringer's lactate solution, and the like. Methods for encapsulating the composition (such as in coatings of hard gelatin or cyclodextrin) are known in the art (Baker et al., "Controlled Release of Biological Active Agents", John Wiley and Sons, 1986). rolled Release of Biological Active Agen ts", John Wiley and Sons, 1986).

[0043] As used herein, the phrase "disorder responsive to modulation of the melanocortin-4 receptor" refers to any disorder that can be treated by activation (stimulation) or inhibition of the MC4R. Examples of such disorders are described in detail below.

[0044] As used herein, the term "modulatory substance" refers to a compound that affects a biological function and interacts with a target receptor. Examples of modulatory substances include full agonists, partial agonists, neutral antagonists, and inverse agonists.

[0045] As used herein, the term "agonist" refers to any natural or synthetic chemical compound that, when it interacts with its target, here the MC4R (e.g., binds to it), raises the signaling activity of the MC4R above its basal level. An agonist can be a superagonist (i.e., a compound that can produce a greater maximum response than the endogenous agonist for the target receptor and thus has an efficacy greater than 100%), a compound that can produce a greater maximum response than the endogenous agonist for the target receptor and A full agonist (i.e., a compound that induces a maximal response after receptor occupancy and activation) ) or a partial agonist (i.e., a compound that can activate the receptor but cannot induce the maximal response of the receptor system) may be used. Examples of MC4R agonists are described in detail below.

[0046] As used herein, the term "antagonist" refers to any chemical compound that, when interacting with its target, here MC4R, (e.g., binding) blocks the signal transduction activity of the agonist compound with the MC4R in a dose-dependent manner. As used herein, the term "inverse agonist" refers to any chemical compound that, when interacting with its target, here MC4R, (e.g., binding) decreases the basal level of the signal transduction activity of MC4R in a dose-dependent manner.

[0047] As used herein, "effective amount" refers to an amount of a therapeutic agent or combination of therapeutic agents sufficient to treat or prophylactically treat a target disorder. Examples of effective amounts typically range from about 0.0001 mg / kg body weight to about 500 mg / kg body weight. One exemplary range is from about 0.0001 mg / kg body weight to about 500 mg / kg. For example, an effective amount may range from about 0.005 mg / kg to about 500 mg / kg. In other examples, the range may be from about 0.0001 mg / kg to about 5 mg / kg. In yet other examples, an effective amount may range from about 0.01 mg / kg body weight to 50 mg / kg body weight, or from 0.01 mg / kg body weight to 20 mg / kg body weight.

[0048] As used herein, the term "subject" refers to a mammal, preferably a human, but examples include veterinary treatment of companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.) and laboratory animals (e.g., rats, mice, guinea pigs, etc.) that may also require

[0049] As used herein, the term "second agent" includes any active pharmaceutical ingredient (API) that, when combined with the peptides described herein, enhances the therapeutic effect produced by the peptides described herein alone, or exhibits a synergistic effect with the peptides described herein (i.e., shows a combined effect greater than an additive effect). As used herein, "enhanced therapeutic effect" includes a therapeutic profile improved by methods other than synergistic effects. Examples of enhanced therapeutic effects include reducing the effective dosage of the peptides described herein, extending the therapeutic window of the peptides described herein, etc. One or more second agents may be administered. Examples of the second agent are described in detail below.

[0050] The second agent may be administered before, simultaneously with, or after the administration of the peptides described herein. Thus, the peptides described herein and the second agent may be administered together as one formulation, or in separate formulations, e.g., simultaneously or sequentially. For example, the peptides described herein and the second agent may be sequentially administered in separate compositions, and the peptides described herein may be administered before or after the second therapeutic agent. In addition, the It may or may not be administered orally. For example, the peptide described herein and the second therapeutic agent may be such that the peptide described herein is administered more frequently than the second therapeutic agent or vice versa, such that the second therapeutic agent is administered more frequently than the peptide described herein and may have different half-lives and / or act on different timescales. Finally, a second agent may be administered after the peptide described herein, and the sequential application of both therapeutic agents may further enhance the therapeutic efficacy Either the peptide or the second agent described herein may be administered acutely or chronically .

[0051] In the method or composition of the present invention, an effective amount can be achieved by administering together a compound having MC4R modulator activity in a first amount or a pharmaceutically acceptable salt thereof and a second amount of at least one second agent . In one embodiment, the peptide described herein and the second agent are each administered in an effective amount (i.e., in an amount that is therapeutically effective when administered alone) . In another embodiment, the peptide described herein and the second agent are each administered in an amount that does not provide a therapeutic effect (less than the therapeutic dosage) . In yet another embodiment, the peptide described herein may be administered in an effective amount, while the second agent is administered in an amount less than the therapeutic dosage . In yet another embodiment, the peptide described herein may be administered in an amount less than the therapeutic dosage , while the second agent is administered in an effective amount. In an exemplary embodiment, the combination of the peptide and the second agent described herein is such that the peptide described alone In an exemplary embodiment, the combination of the peptide and the second agent described herein is such that the peptide described alone In an exemplary embodiment, the combination of the peptide and the second agent described herein is such that the peptide described alone In an exemplary embodiment, the combination of the peptide and the second agent described herein is such that the peptide described alone and the second agent are each less effective than when administered in combination Exhibits an enhanced therapeutic effect or a synergistic effect as compared to either the first or second agent is shown.

[0052] The presence of a synergistic effect can be determined using an appropriate method for evaluating drug interactions which can be. Appropriate methods include, for example, the Sigmoid-Emax equation (Holford, N.H.G. and Scheiner, L.B., Clin Pharmacokine t. 6:429-453 (1981)), the Loewe additivity equation (Loewe, S. and Muischnek, H., Arch. Exp. Pathol. Pharmacol. 114:313-326 (1926)) and the median effect equation (Chou, T.C. and Talalay, P., Adv. Enzyme Regul. 22:27-55 (1 984)). Each of the equations referenced above can be applied to experimental data to create the corresponding graphs to assist in evaluating the combined effect of the drugs which can be. The corresponding graphs associated with the equations referenced above are, respectively, concentration-effect curves, isobologram ram curves and combination index curves are.

[0053] As used herein, "treating" includes achieving partial or substantial delay, inhibition or prevention of the progression of clinical symptoms associated with the target disorder For example, "treating" includes partially or completely reducing body weight (such as as measured by body mass index BMI); improving or ameliorating clinical symptoms or markers associated with obesity, such as . For example, "treating" includes partially or completely reducing body weight (such as as measured by body mass index BMI); improving or ameliorating clinical symptoms or markers associated with obesity, such as ) reducing body weight partially or completely; improving or ameliorating clinical symptoms or markers associated with obesity, such as type II diabetes, prediabetic state, blood levels of hemoglobin Alc (HblAc) higher than 6%, hyperinsulinemia, hyperlipidemia, insulin resistance, glucose intolerance, etc. levels, hyperinsulinemia, hyperlipidemia, insulin resistance, glucose intolerance, etc. slowing, inhibiting, or preventing the progression of obesity and symptoms associated with obesity; or partially or completely delaying, inhibiting, or preventing the occurrence or progression of obesity or symptoms associated with obesity and achieving in part or substantially one or more of the results thereof. Slowing, inhibiting, or preventing the progression of obesity includes, for example, slowing, inhibiting, or preventing the progression from normal weight to obesity in a subject. The term "treating" further includes partially or completely reducing the risk of coronary artery disease, stroke, and diabetes (e.g., type 2) associated with metabolic syndrome, and alleviating or improving clinical symptoms associated with metabolic syndrome or signs of metabolic syndrome, such as any of the five parameters listed above. For example, the term "treating" includes insulin resistance, parameters associated with metabolic syndrome including sucrose craving, and cardiovascular disease parameters including heart rate and blood pressure, delaying, inhibiting, or preventing the progression of diseases of the joints, inflammation, sleep apnea, hyperphagia, other eating disorders including bulimia, adjuvant therapy for weight loss surgery, and adjuvant weight loss treatment before plastic surgery. "Preventive treatment" refers to treatment to prevent, inhibit, or reduce the occurrence of clinical symptoms of the target disorder before they occur. (Disorders Responsive to Modulation of MC4R) Examples of disorders responsive to modulation of MC4R include acute and chronic inflammatory diseases such as systemic inflammation, inflammatory bowel disease, brain inflammation, sepsis, and septic shock; diseases with autoimmune components such as rheumatoid arthritis, gouty arthritis, and multiple sclerosis; diseases associated with weight gain

[0054] (Disorders Responsive to Modulation of MC4R) Examples of disorders responsive to modulation of MC4R include acute and chronic inflammatory diseases such as systemic inflammation, inflammatory bowel disease, brain inflammation, sepsis, and septic shock; diseases with autoimmune components such as rheumatoid arthritis, gouty arthritis, and multiple sclerosis; diseases associated with Metabolic diseases and medical conditions, such as obesity, eating disorders, and Prader-Willi syndrome; metabolic diseases and medical conditions associated with weight loss, such as anorexia, bulimia, AI DS wasting, cachexia, cancer cachexia, and wasting in frail elderly; diabetes and conditions associated with diabetes and diabetes complications, such as retinopathy; neoplastic growths, such as skin cancer and prostate cancer; reproductive or sexual medical conditions, such as endometriosis and uterine bleeding in women, sexual dysfunction, erectile dysfunction, and decreased female sexual response; diseases or conditions resulting from organ treatment or injury, such as organ transplant rejection, ischemia and reperfusion injury, spinal cord injury treatment, and treatments to promote wound healing, and weight loss caused by chemotherapy, radiation therapy, temporary or permanent immobilization, or dialysis; cardiovascular diseases or conditions, such as hemorrhagic shock, cardiogenic shock, hypovolemic shock, cardiovascular disorders, and cardiac cachexia; lung diseases or conditions, such as acute respiratory distress syndrome, chronic obstructive pulmonary disease, asthma, and pulmonary fibrosis; disorders that enhance immune tolerance and fight attacks on the immune system, such as those related to specific allergies or organ transplant rejection; dermatological diseases and conditions, such as psoriasis, decreased skin pigmentation, acne, keloid formation, and treatment of skin cancer; behavioral, central nervous system, or nerve-related conditions and disorders, such as anxiety, depression, memory impairment and memory dysfunction, pain perception regulation, treatment of neuropathic pain; alcohol consumption, alcoholism and / or conditions and diseases associated with alcoholism; and kidney conditions or diseases, such as treatment of kidney cachexia or increased sodium excretion. Further examples include thyroxine release, aldosterone synthesis and release, body temperature, blood pressure, heart rate, vasoconstriction, cerebral blood flow activity or homeostatic activity that normalizes a subject, including blood glucose level, bone metabolism, bone formation or growth, ovary weight, placenta growth, prolactin and FSH secretion, fetal growth in utero, parturition, spermatogenesis, sebum and pheromone secretion, nerve protection and nerve growth, and motivation, learning and regulation of other behaviors. Further examples include bulimia, binge eating or other eating disorders.

[0055] In an exemplary embodiment, a disorder responsive to regulation of the MC4R receptor is type 1 diabetes, type 2 diabetes, obesity, insulin resistance, metabolic syndrome, cardiovascular disease, or low density lipoprotein / high density lipoprotein / triglyceride imbalance, non-alcoholic fatty liver disease and substance abuse disorder.

[0056] In an exemplary embodiment, a disorder responsive to regulation of the MC4R receptor is type 1 diabetes, type 2 diabetes, obesity, insulin resistance or metabolic syndrome.

[0057] (Obesity) As used herein, the term "obesity" refers to a subject with a body mass index (BMI) of about 30 kg / m / m 2 or more, for example, a subject with a BMI of 25, 26, 27, 28, 29, 30, 31, 32, 33 34, 35, 36, 37 kg / m 2 or more. In certain embodiments, an obese subject has a BMI within the range defined as "obesity" by the Center for Disease Control. See URL http: / / www.cdc.gov / obe sity / defining.html (last accessed October 28, 2011 ). For example, in one embodiment, BMI ≧ 30.0 kg / m ) 2The adult is obese .

[0058] (Diabetes and related disorders) In an exemplary embodiment, the subject treated by the method provided by the present invention has a disorder related to diabetes or an increased risk of developing. "Disorders related to diabetes" refers to diabetes (type 1 (OMIM 222100) and type 2 (OMIM 125 853)), insulin resistance and metabolic syndrome. In an exemplary embodiment, the subject to be treated has diabetes (type 1 or type 2), insulin resistance , or metabolic syndrome. In an exemplary embodiment, the disorder is diabetes, for example, type 2

[0059] diabetes. In an exemplary embodiment, the subject has type 2 diabetes as defined by the World Health Organ ization and International Diabetes Federat ion in "Definition and diagno sis of diabetes mellitus and intermediat e hyperglycaemia" published in 2006 (the whole of which is incorporated herein by reference. ). In an exemplary embodiment, the diabetic subject has a fasting plasma glucose of 126 mg / dL or more or a 2-hour plasma glucose (2 hours after oral administration of 75 glucose) of 2 00 mg / dL or more. In an exemplary embodiment, the diabetic or prediabetic subject shows a high level of glycated hemoglobin, for example, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.8, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.8, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.8, Above 6.0, 6.2, 6.4, 6.6, 6.8, 7.0, 7.2, 7.4, 7.6% indicates glycated hemoglobin. In an exemplary embodiment, one or more of the genes in Table 1 below or near one or more genes may have a genetic polymorphism (e.g., a polymorphism that results in an altered expression level, e.g., one that increases or decreases the expression level and / or results in a variation in the coding sequence), by which a diabetic or prediabetic subject may be identified or further characterized. [Table 2]

[0060] In an exemplary embodiment, additional genes that can be used to identify or further characterize a subject treated by the method provided by the present invention include FTO ( OMIM 610966), JAZF1 (OMIM 606246) and HHEX ( OMIM 604420).

[0061] In an exemplary embodiment, the subject treated by the method of the present invention has type I diabetes. In an exemplary embodiment, a subject having type I diabetes can be determined by a C-peptide assay, e.g., the fasting C-peptide level is less than about 1.0 nmol / L, e.g., less than 1.2 nmol / L, less than 1.1 nmol / L, less than 1.0 nmol / L, less than 0.9 nmol / L, less than 0.8 nmol / L, less than 0.7 nmol / L, less than 0.6 nmol / L, less than 0.5 nmol / L, less than 0.4 nmol / L, or even less, e.g., less than 0.33 nmol / L, less than 0.25 nmol / L, less than 0.2 nmol / L, or . It is characterized by being less than 0.1 nmol / L. In an exemplary embodiment, C-peptide levels are measured after an oral glucose challenge (administering 75 g of glucose and measuring after 2 hours), and an increase of less than 0.54 nmol / L, for example, less than 0.50 nmol / L, 0.45 nm ol / L, less than 0.40 nmol / L, less than 0.35 nmol / L, less than 0.30 nmo l / L, less than 0.25 nmol / L, less than 0.20 nmol / L, less than 0.15 nmol / L, or less than 0.10 nmol / L is detected. Abnormal fasting blood glucose levels (1 10 - 125 mg / dL) or impaired glucose tolerance (2 hours after a 75 g glucose challenge : 140 - 199 mg / dL) can be used to identify or further characterize the decline in β-cell function in subjects with type 1 diabetes. In an exemplary embodiment, type 1 diabetes is identified or further characterized by the presence of autoantibodies against islet cell antigens and / or insulin, such as the 65 kDa GAD (OMIM 138275) and / or autoantibodies associated with the IA-2 molecule related to phosphatase

[0062] (insulin resistance) In an exemplary embodiment, the disorder is "insulin resistance" and may be identified by any means known in the art, and is characterized by a decrease in the ability of insulin to lower blood glucose levels. In an exemplary embodiment, insulin resistance is associated with RETN, PTPN1, TCF1 (OMIM 142410; see, for example, polymorphism 0011), PPP1R3A (OMIM 60 0917; see, for example, polymorphisms 0001, 0003), PTPN1 (OMIM 1768 85; see, for example, polymorphism 0001), ENPP1 (OMIM 173335; for example, Refer to polymorphism 0006), IRS1 (OMIM 147545; e.g., refer to polymorphism 0002) ), EPHX2 (OMIM 132811; e.g., refer to polymorphism 0001), leptin (OMIM 164160, e.g., refer to polymorphisms 0001 and 0002), leptin receptor (OMIM 601007, e.g., refer to polymorphisms 0001, 0002, 0004, and 00 05), or the presence of one or more polymorphisms of one or more genes of the insulin receptor (INSR, OMIM 147670, e.g., refer to polymorphisms 0001 - 0037) is determined or further characterized by ( e.g., polymorphisms that result in altered expression levels, e.g., increasing or decreasing the expression level, and / or polymorphisms of the coding sequence of the gene product, e.g., polymorphisms that result in variations in the protein).

[0063] (Metabolic syndrome) In an exemplary embodiment, the disorder is metabolic syndrome. As used herein, the term "metabolic syndrome" refers to a group of symptoms in which coronary artery disease, stroke, and type 2 diabetes occur together and the risks of these increase. According to the American Heart Association and the National Heart, Lung and Blood In stitute, metabolic syndrome, also known as syndrome X, is present when a subject has three or more of the following signs.

[0064] (1) Blood pressure of 130 / 85 mmHg or higher, (2) Fasting blood glucose (glucose) of 100 mg / dL or higher, (3) Large waist circumference (length around the waist): - For men - 40 inches or more - For women - 35 inches or more (4) Low HDL cholesterol: - Male - Lower than 40 mg / dL - Female - Lower than 50 mg / dL (5) Triglyceride is 150 mg / dL or higher.

[0065] The metabolic syndrome can be diagnosed by testing the subject's blood pressure, blood glucose level, HDL cholesterol level, LDL cholesterol level, total cholesterol level, and triglyceride level. It can be done.

[0066] In an exemplary embodiment, the subject has central obesity (in the case of females, waist circumference ≥ 8 0 cm; in the case of Asian males including South Americans and Central Americans, ≥ 90 cm, and in all other males, ≥ 94 cm), BMI > 30 kg / m 2 , elevated triglycerides (≥ 150 mg / dL, or specific treatment for this lipid abnormality), reduced HDL cholesterol ( in males, < 40 mg / dL, in females, < 50 mg / dL, or specific treatment for this lipid abnormality), elevated blood pressure (sBP ≥ 130 mmHg or dBP ≥ 85 mmHg, or treatment for already diagnosed hypertension), or elevated fasting plasma glucose (FPG ≥ 100 mg / dL, or previous diagnosis of type 2 diabetes), including combinations thereof. In an exemplary embodiment , the subject treated by the method provided by the present invention has or is at risk of having the metabolic syndrome as defined by the "T he IDF consensus worldwide definition of the metabolic syndrome" published by the International Diabetes Federation in 2006 (the whole of which is incorporated herein by reference and is incorporated herein by reference in its entirety), as defined therein. or risk of metabolic syndrome is increasing. That is, the subject has central obesity (as described above and / or BMI > 30 kg / m 2 ), and has any two of an increase in triglycerides, a decrease in HDL cholesterol , an increase in blood pressure, or an increase in fasting plasma glucose. In an example embodiment, metabolic syndrome is characterized in the subject by the presence of a mutation at locus 3q27 (see, e.g., OMIM 605 552) and / or 17p12 (see, e.g., OMIM 605572), or is further characterized.

[0067] (Disorders caused by MC4R mutations) The present invention relates to a method of treating a disorder in a subject resulting from a weakened response of MC4R to the hormone hormone that stimulates α-melanocortin (α-MSH). This method comprises administering an effective amount of a melanocortin-4 receptor (MC4R) agonist. In an example embodiment, the subject is a heterozygous carrier of an MC4R mutation that weakens the response of MC4R to the hormone hormone that stimulates α-melanocortin (α-MSH). Since heterozygous owners retain the ability to respond to the natural ligand of MC4R, treatment of disorders

[0068] associated with MC4R by administration of an MC4R agonist in heterozygous carriers does not depend on knowledge of the type of MC4R mutation.

[0069] The human MC4R gene (hMC4R) has the GenBank accession number CH471077. MC4R is a well-characterized protein encoded by the genomic sequence Mutations in the receptor are a related cause of severe childhood obesity. The prevalence of MC4R mutations and carriers in this population has been shown to be approximately 2.5%, with severe Among children with obesity, the highest prevalence was 6%. described mice carrying more or less mutations in the MC4 receptor gene. These individuals exhibit a phenotype similar to that of the control group. Glandular and sex steroid levels were unchanged, while lean body mass, bone mineral density, and linear growth rate were The severity of the condition is often accompanied by hyperphagia, hyperinsulinemia, and increased fat mass. In contrast to the deletion, hyperphagia and hyperinsulinemia increase with age in human subjects. As in MC4R knockout mice, the expression of heterozygotes is The type is moderate compared to homozygotes. The hyperphagia observed was less severe than that observed in leptin-deficient humans. The severity of MC4 receptor dysfunction as seen in the ro assay may explain this particular mutation. The amount of food consumed by a subject with a disorder in a test meal can be predicted, and the onset of an obesity phenotype and At least 90 different MC4 receptor mutations have been linked to obesity. It is possible that additional mutations in the MC4 receptor could result in a similar obesity phenotype. Highly sexual.

[0070] An example of an MC4R mutation causing obesity in humans is Farooqi et al., The Journal of Clinical Investigation, 2000 July, vol.106(2), pp.271 - 279 and Vaisse et al., The J ournal of Clinical Investigation, July 2000 , vol.106(2), pp.253 - 262, the relevant parts of which are incorporated herein by reference.

[0071] Additional mutations that potentially cause obesity in humans include Xiang et al., " Pharmacological characterization of 30 h uman melanocortin - 4 receptor polymorphis ms with the endogenous proopiomelanocort in - derived agonists, synthetic agonists a nd the endogenous agouti - related protein antagonist". Biochemistry, June 8, 2010;49(2 2):4583 - 600 (the relevant parts of which are incorporated herein by reference) describes that R18H, R18L, S36Y, P48S, V50M, F51L, E6 1K, I69T, D90N, S94R, G98R, I121T, A154D, Y157S , W174C, G181D, F202L, A219V, I226T, G231S, G23 8D, N240S, C271R, S295P, P299L, E308K, 1317V, L 325F and 750DelGA are included.

[0072] Examples of additional mutations that potentially cause obesity in humans are Online Me Human genes in Mendelian Inheritance in Man (OMIM) and data on genetic disorders, available through the database at http: / / omim.org / entry / 155541, accession number 155541 (MC4R) (more precisely, accession numbers 155541.0001 to 155541.0023). Representative examples include 4-BP DEL, NT631; 4-BP INS, NT732; TYR35TER; ASP37VAL; SER58CYS; ILE102SER; ASN274SER; 1-BP INS, 112A; 4-BP DEL, 211CTCT; ILE125LYS; ALA175THR; ILE316SER; TYR287TER; ASN97ASP; 15-BP DEL (delta 88-92 codons); and SER127LEU. The relevant portions of the OMIM database are incorporated herein by reference.

[0073] In an exemplary embodiment, the mutation of MC4R retains the signaling activity of MC4R.

[0074] Mutations in the genomic sequence encoding MC4R can be detected by methods well known to those skilled in the art. For example, the genomic sequence can be amplified using nucleotide primers such as those described by Farooq i et al., The Journal of Clinical Investigation, July 2000, vol.106(2), pp.271-279 and Vaisse et al., The Journal of Clinical used for cloning, and the cloned array can be analyzed using a commercially available sequencer and software can be analyzed using a commercially available sequencer and software

[0075] The activity of MC4R can be measured by methods well known to those skilled in the art. For example, clo ning the MC4R DNA, transiently transfecting cells with the cloned DNA, and contacting the transfected cells with an MC4R agonist (e.g., α-MSH), and measuring the intracellular level of cAMP, which is the second messenger of MC4R, by, for example, the electrochemiluminescence assay determined by Roubert et al., Journal of Endocrinology (2010) 207, pp. 17 7 - 183. A decrease in MC4R signal transduction can be confirmed by comparing the intracellular level of cAMP produced in response to a given agonist by wild-type C4R with the intracellular level produced by mutant MC4R.

[0076] MC4R modulators (e.g., agonists) can be used to treat patients suffering from other disorders, including a decrease in the availability of the natural agonist of MC4R. Examples of such patients include individuals who are heterozygous or homozygous for mutations in genes important in the leptin-dependent pathway or proopiomelanocortin processing (Nature Clinical Practice Endocrinology and Met abolism, 2006; 2; 6; 318 and N Eng J Med: 2007; 3 56; 3; 237, Nature Genetics, 1998, 155; Cell M etabolism, 2006; 3; 135; Annals Acad Med, 200 et al., Nature Genetics, 1998, 155; Cell Metabolism, 2006; 3; 135; Annals Acad Med, 200 ​9, 38; 1; 34), or individuals having a mutation in the gene encoding prohormone convertase include.

[0077] (Mode of administration) The compounds of the present invention useful for practicing the methods described herein, or pharmaceutically acceptable salts thereof, or administration of the compound or pharmaceutical salt may be continuous, every hour, once a day 4 times, 3 times a day, 2 times a day, once a day, once every other day, twice a week, once a week, 2 once every two weeks, once a month, or once every two months, or for a longer period or some other intermediate dosing schedule may be used.

[0078] Examples of administration of the compounds of the present invention, or compositions containing the compounds of the present invention or pharmaceutical salts include peripheral administration. Examples of peripheral administration include oral, subcutaneous, intraperitoneal, intramuscular, intravenous intravenous, rectal, transdermal, oral, sublingual, inhalation, pulmonary, or nasal administration forms.

[0079] (Combination therapy) To treat any disorder responsive to the regulation of MC4R, by administering in combination with one or more other pharmaceutically active compounds (the "second agent"), the peptides described herein can be used. Such combination administration can be carried out by a single dosing containing one or more peptides described herein and one or more second agents described herein, and, for example, such single dosing forms include tablets, capsules, sprays, inhalation powders inhalation powders, injectable liquids, etc. Alternatively, combination administration can be carried out by two different dosing forms and one dosing form contains one or more peptides described herein and the other dosing form contains one or more second agents, and can be carried out by administering the two dosing forms at different times and one dosing form contains one or more peptides described herein and other dosage forms contain one or more second drugs. In this case, the dosage forms contain the same and may be different. Without being meant to limit the combination therapy, The following are examples of specific combination therapies that may be used:

[0080] The peptides described herein can be used to treat a variety of weight and feeding related disorders, such as obesity, and / or overweight. In particular, the second drug inhibits energy expenditure, glycolysis, gluconeogenesis, glycogenolysis, lipolysis, Fat production, fat absorption, fat storage, fat excretion, hunger and / or satiety and / or cravings were anti-obesity drugs that affect the mechanism of obesity, appetite / motivation, food intake, or gastrointestinal motility. Drugs that reduce energy intake include, in part, various pharmacological agents ( These include anorectic drugs, which are adjuncts to behavioral treatments in weight loss programs. Used as a substance.

[0081] Generally, when used in combination with one or more peptides described herein The total dosage of the obesity control agent or medicine is 0.1 to 3,000 mg / day, preferably about 1 ~1,000 mg / day, more preferably about 1 to 200 mg / day in one or two to four doses. However, the actual dosage is determined and administered by the attending physician. The efficacy of the compound administered will vary depending on factors such as the age, weight, condition and response of the patient.

[0082] One or more of the peptides described herein may be administered to one or more of the target peptides useful for the treatment of diabetes. May be combined with drug 2.

[0083] One or more of the peptides described herein, in addition to, or instead of, diseases, disorders and / or conditions associated with obesity and / or overweight (e.g., insu lin resistance; impaired glucose tolerance; type 2 diabetes; metabolic syndrome; dyslipidemia (including hyperlipidemia); hypertension ; heart disorders (e.g., coronary heart disease, myocardial infarction); cardiovascular disorders; non-alcoholic fatty liver disease (including non-alcoholic steatohepatitis); joint disorders (including secondary arthritis); gastroesophageal reflux ; sleep apnea; atherosclerosis; stroke; large and small blood vessel diseases; lipomatosis (e.g of the liver); gallstones; and gallbladder disorders) may be further combined with one or more second agents useful for the treatment of .

[0084] (Second agent) One or more second agents are selected from, for example, the following.

[0085] Insulin and insulin analogs; Sulfonylureas (e.g., glypidide) and dietary glucose regulators (sometimes referred to as "short-acting secretagogues"), e.g., meglitinides (e.g., repaglinide and nateglinide), insulin secretagogues; Agents that enhance the action of incretin: incretin, incretin mimetics, agents that enhance incretin function, e.g., GLP-1, GIP; GLP-1 agonists (e.g exenatide and liraglutide (VICTOZA)), DPP-4 inhibitors (e.g vildagliptin, saxagliptin and sitagliptin); Peroxisome proliferator-activated receptor gamma (PPARγ) agonists, e.g., thiazo lidinediones (e.g., pioglitazone and rosiglitazone) and PPARα, γ An insulin sensitizer comprising an agent having any combination of the activities of Agents that regulate glucose homeostasis in the liver, such as biguanides (e.g., metformin); Fructose 1,6-bisphosphatase inhibitors, glycogen phosphorylase inhibitors; Glycogen synthase kinase inhibitors and glucokinase activators; Agents designed to reduce / delay glucose absorption from the intestine, such as α-glucosidase inhibitors (e.g., miglitol and acarbose); Agents that antagonize the action of glucagon or reduce glucagon secretion, such as amylin analogs (e.g., pramlintide); Agents that prevent renal glucose reabsorption, such as sodium-dependent glucose transporter 2 (SGLT-2) inhibitors (e.g., dapagliflozin); Agents designed to treat complications of long-term hyperglycemia, such as aldose reductase inhibitors (e.g., epalrestat and ranirestat); Agents used to treat complications associated with microangiopathy; Anti-lipid disorder agents, such as HMG-CoA reductase inhibitors (statins, e.g., rosuvastatin) and other cholesterol-lowering agents; PPARα agonists (fibrates, e.g., gemfibrozil and fenofibrate); Bile acid sequestrants (e.g., cholestyramine); Cholesterol absorption inhibitors (e.g., plant sterols (i.e., phytosterols), synthetic inhibitors); Cholesteryl ester transfer protein (CETP) inhibitors; inhibitors of the ileal bile acid transport system (IBAT inhibitors); Resins to which bile acids bind; Nicotinic acid (niacin) and its analogs; ; ; ; ; ; inhibitors of the ileal bile acid transport system (IBAT inhibitors); ; Resins to which bile acids bind; Nicotinic acid (niacin) and its analogs; Antioxidants, such as probucol; ω-3 fatty acids; Adrenergic receptor antagonists, such as β-blockers (e.g., atenolol), α-blockers (e.g., doxazosin), and mixed α / β-blockers (e.g., labetalol), for lowering blood pressure; α-2 agonists (e.g., clonidine), adrenergic receptor agonists; 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-acting adrenergic agents, such as central α-agonists (e.g., clonidine); And diuretics (e.g., furosemide); Antithrombotic agents, such as fibrinolysis activators; hemostatic modifiers containing thrombin antagonists; Factor VIIa inhibitors; anticoagulants, such as vitamin K antagonists (e.g., warfarin), heparin and its low molecular weight analogs, factor Xa inhibitors, and direct thrombin inhibitors (e.g., argatroban); antiplatelet agents, such as cyclooxygenase inhibitors (e.g., aspirin), adenosine diphosphate (ADP) receptor inhibitors (e.g., clopidogrel), phosphodiesterase inhibitors (e.g., cilostazol), glycoprotein IIb / IIIa inhibitors (e.g., tirofiban), and adenosine reuptake inhibitors (e.g.,); ; Angiotensin II receptor antagonists (e.g., candesartan); aldosterone receptor antagonists (e.g., eplerenone); ; Central-acting adrenergic agents, such as central α-agonists (e.g., clonidine); And diuretics (e.g., furosemide); Antithrombotic agents, such as fibrinolysis activators; hemostatic modifiers containing thrombin antagonists; ; Factor VIIa inhibitors; anticoagulants, such as vitamin K antagonists (e.g., warfarin), heparin and its low molecular weight analogs, factor Xa inhibitors, and direct thrombin inhibitors (e.g., argatroban); antiplatelet agents, such as cyclooxygenase inhibitors (e.g., aspirin), adenosine diphosphate (ADP) receptor inhibitors (e.g., clopidogrel), phosphodiesterase inhibitors (e.g., cilostazol), glycoprotein IIb / IIIa inhibitors (e.g., tirofiban), and adenosine reuptake inhibitors (e.g.,); ; ; ; ; ; For example, dipyridamole); Anti-obesity agents, such as noradrenergic agents (e.g., phentermine) and sero tonergic agents (e.g., sibutramine), pancreatic lipase inhibitors (e.g., orlistat ), microsomal transport protein (MTP) modifiers, diacylglycerol acyltra nsferase (DGAT) inhibitors and cannabinoid (CB1) receptor antago nists (e.g., rimonabant), appetite suppressants (e.g., ephedrine); Feeding behavior regulators, such as orexin receptor modifiers and melanin-concentrating hormone (M CH) modifiers; Neuropeptide Y (NPY) / NPY receptor modifiers; Pyruvate dehydrogenase kinase (PDK) modifiers; Serotonin receptor modifiers; Leptin / leptin receptor modifiers; Ghrelin / ghrelin receptor modifiers; Agents that enhance the function of β-cells; Agents that stimulate energy consumption (e.g., β-adrenergic stimulants, UCP-1 ago nists, brown fat modifiers and stimulants); Agents that induce lysis of adipocytes (e.g., antibodies); Nicotine or nicotine withdrawal aids; Estrogen, natural or synthetic modifiers of estrogen receptors; μ-opioid receptor modifiers; and Monoamine transmission regulators, such as selective serotonin reuptake inhibitors (SSRI)( e.g., fluoxetine), noradrenaline reuptake inhibitors (NARI), norad renaline-serotonin reuptake inhibitors (SNRI), triple monoamine reuptake blo ckers (e.g., tesofensine) and monoamine oxidase inhibitors (MAOI)(e.g For example, troxaton and amiflamine, or a pharmaceutically acceptable salt thereof.

[0086] In an exemplary embodiment, the MC4R agonist and the second agent are administered simultaneously, sequentially, or separately with a very low calorie diet (VLCD) or a low calorie diet (LCD).

[0087] (the isolated polypeptide of the present invention) In an exemplary embodiment, the isolated polypeptide (e.g., an agonist of MC4R) is , the formula (I) or a pharmaceutically acceptable salt thereof.

[0088] The isolated polypeptide of the following structural formula (I):

Chemical formula

[0089] Or a pharmaceutically acceptable salt thereof, Wherein, R 1 Is H, or C1-C6 acyl; R 2 Is -NR 3 R 4 Or -OR 5 And R 3 , R 4 And R 5 Are each independently And is H or C1-C6 alkyl; A 1 Is an amino acid residue selected from Arg, Lys, Orn, His, Nle, Phe, Val, Leu, Trp , Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzAl Selected from a; or A 1 Is optionally substituted C1-C12 alkyl, optionally substituted C6-C 18 aryl, optionally substituted C5-C18 heteroaryl, where the aryl moiety is substituted C6-C18 aryl, optionally substituted, The aralkyl, 1-C12 alkyl, or heteroaryl moiety may be substituted. C5-C18 heteroaryl, the alkyl moiety of which is optionally substituted, heteroaralkyl, wherein the heteroaryl is aryl; A 2 and A 8 are independently Cys, hCys, Pen, Asp, Glu, is an amino acid residue selected from Lys, Orn, Dbu, or Dpr; 2 and A 8 are selected such that, in pairs, a covalent bond can be generated between each of the side chains; A 3 is absent or Ala, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib or residue Y, wherein Y is Structural formula of [ka]

[0090] and wherein the amino acid is selected from the amino acids represented by R 11 and R 12 are each independently H, -CH3, phenyl or benzyl. can be; R 21 , R 22 , R 23 and R 24 are each independently H, -CH3, or -CF3 , phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; R 31 , R32 , R 33 , R 34 , R 41 , R 42 and R 43 are each independently , H, -CH3, -CF3, phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; A 4 is absent or is an amino acid residue selected from Atc, Ala, Q Ala, Aib, Sar, Ser , Thr, Pro, Hyp, Asn, Gln, optionally substituted His, Trp, Tyr , Lys, Arg, sChp, or residue X, where X is an amino acid selected from the amino acids represented by the following structural formula

Chemical formula

[0091] and is wherein R 51 and R 52 are each independently H, -CH3, phenyl, or benzyl ; R 61 , R 62 , R 63 and R 64 are each independently H, -CH3, -CF3 , phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; R 71 , R 72 , R 73 , R 74 , R 81 , R 82 and R 83 are each independently , H, -CH3, -CF3, phenyl, benzyl, F, Cl, Br, I, -OCH3, or -OH; A 5 is optionally substituted Phe, optionally substituted 1-Nal, or optionally substituted 2-Nal; A 6 is Arg; A 7 is Trp, and any amino acid residue is in either the L-configuration or the D-configuration.

[0092] In an exemplary embodiment, A 3 is absent or is an amino acid selected from Ala, Tle, Val, Le u, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gl n, Sar, Gly, Asn, or Aib; A 4 is absent or is an amino acid residue selected from Atc, Ala, QAla, Aib, Sar, Ser, Thr, Pr o, Hyp, Asn, Gln, optionally substituted His, Trp, Tyr, Lys, Arg, sChp, or residue X, where X is an amino acid selected from the amino acids represented by the following structure formula formula

Chemical formula

[0093] as represented by.

[0094] The remaining values and preferred values of the variables are as defined above and below for formula (I) respectively.

[0095] In an exemplary embodiment, A 3 and A 4 are each independently the following structural formula

Chemical formula

[0096] is a residue of an amino acid selected from the amino acids represented by

[0097] The remaining values and preferred values of the variables are as defined above and below for formula (I) respectively.

[0098] In an exemplary embodiment, A 3 and A 4 are both not absent. The remaining values and preferred values of the variables are as defined above and below for formula (I).

[0099] In an exemplary embodiment, when A 4 is an amino acid, A 3 is neither Aib nor Gly . The remaining values and preferred values of the variables are as defined above and below for formula (I) respectively.

[0100] In an exemplary embodiment, when A 4 is His and A 5 is D-Phe or 2-Nal , A 3 is neither a D-amino acid nor L-Ala. The remaining values and preferred values of the variables are as defined above and below for formula (I).

[0101] In an exemplary embodiment, when A 2 and A 8 are each selected from Cys, hCys or Pen , (a) when A 4 is absent, A 3 is not L-His; (b ) when A 3 is absent, A 4 is not L-His; (c) when A 4 is His , A 3is not Glu, Leu, or Lys. The remaining values and preferred values of the variables are as defined above and below for formula (I).

[0102] In an exemplary embodiment, (1) A 3 and A 4 are both not absent; (2) when A 4 is an amino acid, A 3 is neither Aib nor Gly; (3) when A 4 is His and A 5 is D-Phe or 2-Nal, A 3 is neither a D-amino acid nor L-Al a; (4) when A 2 and A 8 are each selected from Cys, hCys, or Pen respectively, (a) when A 4 is absent, A 3 is not L-His; (b) when A 3 is absent, A 4 is not L-His; (c) when A 4 is His, A 3 is not Glu, Leu, or Lys. The remaining values and preferred values of the variables are as defined above and below for formula ( I).

[0103] In another embodiment, for the polypeptide of formula (I), A 4 is an L-amino acid. In yet another embodiment, A 4 is absent. The remaining values and preferred values of the variables are as defined above and below for formula (I).

[0104] In an exemplary embodiment, A 5 is optionally substituted 1-Nal or optionally substituted ​It may also be 2-Nal, for example, it may be D-2-Nal which may be substituted. A 5 is any of 5 aromatic carbons and may be substituted with a substituent selected from F, CI, Br, I, -CH3, -OH, -CN , amine, -NO2, or -OCH3.

[0105] In a further embodiment, the polypeptide of formula (I), A 5 is optionally substituted D- Phe. A 5 is any of 5 aromatic carbons and may be substituted with a substituent selected from F, CI, Br, I, -CH3 , -OH, -CN, amine, -NO2, or -OCH3. It may be substituted. A 5 Suitable examples of include, but are not limited to, Phe, Phe(2’-F ), Phe(2’-Cl), Phe(2’-Br), Phe(2’-I), Phe(2’ -CN), Phe(2’-CH3), Phe(2’-OCH3), Phe(2’-CF3 ), Phe(2’-NO2), Phe(3’-F), Phe(3’- Cl), Phe( 3’-Br), Phe(3’-I), Phe(3’-CN), Phe(3’-CH3), Phe(3’-OCH3), Phe(3’-CF3), Phe(3’-NO2), Phe (4’-F), Phe(4’-Cl), Phe(4’-Br), Phe(4’-I), P he(4’-CN), Phe(4’-CH3), Phe(4’-OCH3), Phe(4 ’-CF3), Phe(4’-NO2), Phe(4’-t-Bu), Phe(2’,4 ’-diF), Phe(2’,4’-diCl), Phe(2’,4’-diBr), Phe( 2’,4’-diI), Phe(2’,4’-di-CN), Phe(2’,4’-di-CH 3), Phe(2’,4’-di-OCH3), Phe(3’,4’-diF), Phe(3 ’,4’-diCl), Phe(3’,4’-diBr), Phe(3’,4’-diI), P he(3’,4’-di-CN), Phe(3’,4’-di-CH3), Phe(3’,4 ’-di-OCH3), Phe(3’,5’-diF), Phe(3’,5’-diCl), P he(3’,5’-diBr), Phe(3’,5’-diI), Phe(3’,5’-di- CN), Phe(3’,5’-diCH3), Phe(3’,5’-di-OCH3), and also include D-amino acids selected from Phe(3’,4’,5’-triF). The remaining values and preferred values of the variable are as defined above and below for formula (I). .

[0106] In a further embodiment, the polypeptide of formula (I), A 5 is optionally substituted D- 2-Nal. 5 A is at any of the 5 aromatic carbons and is optionally substituted with a substituent selected from F, CI, Br, I, -C H3, -OH, -CN, amine, -NO2, or -OCH3.

[0107] In yet another embodiment, the polypeptide of formula (I), A 4 is His optionally substituted at any suitable position with a substituent selected from F , CI, Br, I, -CH3, -OH, -CN, amine, -NO2, benzyl or -O CH3. The remaining values and preferred values of the variable are as defined above and below for formula (I).

[0108] In certain embodiments, the compounds of the invention have an EC 50 (MC1R) / E ​​C 50 (MC4R) while having a ratio to MC4R of EC 50 is about 0.01 nM to about 10 nM, for example, 0.01 - 3 nM, and is a polypeptide of formula (I).

[0109] In another embodiment, the polypeptide of the present invention has the following structural formula: [Chemical formula] TIFF2025102830000017.tif34141

[0110] is represented by any of the following polypeptides, or a pharmaceutically acceptable salt thereof.

[0111] In yet another embodiment, the polypeptide of the present invention has the following structural formula: [Chemical formula]

[0112] and includes any of the following, or a pharmaceutically acceptable salt thereof.

[0113] In a further embodiment, the polypeptide of the present invention has the following structural formula: [Chemical formula]

[0114] is represented by any of the following polypeptides, or a pharmaceutically acceptable salt thereof.

[0115] In yet another embodiment, the polypeptide of the present invention includes the polypeptide represented by formula (I), and A is Atc, Ala, Q Ala, Aib, Sar, Ser, Thr, 4 Pro, Hyp, Asn, Gln, substituted His, Trp, Tyr, Lys, Arg Pro, Hyp, Asn, Gln, substituted His, Trp, Tyr, Lys, Arg is an amino acid residue selected from sChp, or residue X. Examples of such peptides include the following structural formulas: Ac-Arg-cyclo[Cys-D-Ala-His(3-Me)-D-Phe-Ar g-Trp-Cys]-NH2; (SEQ ID NO: 36) Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Ar g-Trp-Cys]-NH2; (SEQ ID NO: 37) Ac-Arg-cyclo[Cys-D-Ala-Trp-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 9) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 8) Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 7) Ac-Arg-cyclo[Cys-D-Ala-Arg-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 38) Ac-Arg-cyclo[Cys-D-Ala-Tyr-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 39) Ac-Arg-cyclo[Cys-D-Ala-D-Pro-D-Phe-Arg-Tr p-Cys]-NH2; (SEQ ID NO: 40) Ac-Arg-cyclo[Cys-D-Ala-Pro-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 2) Ac-Arg-cyclo[Cys-D-Ala-Pro-D-Phe(p-F)-Arg -Trp-Cys]-NH2; (SEQ ID NO: 4) Ac-Arg-cyclo[Cys-D-Ala-Atc-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 41) Ac-Arg-cyclo[Cys-D-Ala-QAla-D-Phe-Arg-Trp -Cys]-NH2; (SEQ ID NO: 42) Ac-Arg-cyclo[Cys-D-Ala-sChp-D-Phe-Arg-Trp -Cys]-NH2; (SEQ ID NO: 43), or Ac-Arg-cyclo[Cys-D-Ala-X-D-Phe-Arg-Trp-Cy s]-NH2, (SEQ ID NO: 44), any one of the peptides represented by, or its pharmaceutically acceptable salts.

[0116] In an exemplary embodiment, the polypeptide of the present invention has the following structural formula: Ac-Arg-cyclo[hCys-Ala-D-Phe-Arg-Trp-Cys]- NH2; (SEQ ID NO: 15) Ac-Arg-cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Cys -NH2; (SEQ ID NO: 14) Ac-Arg-cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Pen -NH2; (SEQ ID NO: 45) Ac-Arg-cyclo[Glu-D-Ala-D-Phe-Arg-Trp-Dpr] -NH2; (SEQ ID NO: 26) Ac-Arg-cyclo[Glu-Ala-D-Phe-Arg-Trp-Dpr]-N H2; (SEQ ID NO: 27) Ac-Arg-cyclo[hCys-Aib-D-Phe-Arg-Trp-Cys]- NH2; (SEQ ID NO: 46) Ac-Arg-cyclo[hCys-Sar-D-Phe-Arg-Trp-Cys]- NH2; (SEQ ID NO: 47) Ac-Arg-cyclo[hCys-Val-D-Phe-Arg-Trp-Cys]- NH2; (SEQ ID NO: 48) Ac-Arg-cyclo[hCys-D-Val-D-Phe-Arg-Trp-Cys -NH2; (SEQ ID NO: 49) Ac-Arg-cyclo[hCys-Gln-D-Phe-Arg-Trp-Cys]- NH2; (SEQ ID NO: 50) Ac-Arg-cyclo[hCys-D-Gln-D-Phe-Arg-Trp-Cys -NH2; (SEQ ID NO: 51) Ac-Arg-cyclo[hCys-Ala-D-Phe-Arg-Trp-Pen]- NH2; (SEQ ID NO: 52) Ac-Arg-cyclo[D-Pen-D-Ala-D-Phe-Arg-Trp-hC ys]-NH2; (SEQ ID NO: 53) Ac-Arg-cyclo[Cys-D-Ala-D-Phe-Arg-Trp-hCys -NH2; (SEQ ID NO: 17) Ac-Arg-cyclo[Pen-D-Ala-D-Phe-Arg-Trp-hCys -NH2; (SEQ ID NO: 54) Ac-Arg-cyclo[D-hCys-D-Ala-D-Phe-Arg-Trp-C ys]-NH2; (SEQ ID NO: 55) Ac-Arg-cyclo[hCys-Pro-D-Phe-Arg-Trp-Cys]- NH2; (SEQ ID NO: 20), or Ac-Arg-cyclo[hCys-D-Pro-D-Phe-Arg-Trp-Cys -NH2 (SEQ ID NO: 56), or a polypeptide represented by any one of them, or a pharmaceutically acceptable salt thereof.

[0117] In another embodiment, the polypeptide of the present invention comprises a polypeptide represented by formula (I), wherein A is Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, 3 is Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Selected from Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, or Aib is the selected amino acid; A 4 is an amino acid residue selected from Atc, Ala, QAla, Aib, Sar, Ser, Thr, Pro, Hyp, Asn, Gln, substituted His, Trp, Tyr, Lys, Arg, sChp, or is residue X. Examples of such polypeptides are represented by the following structural formulas: Ac-Arg-cyclo[Cys-Val-Gln-D-Phe-Arg-Trp-Cys]-NH2; (SEQ ID NO: 57) Ac-Arg-cyclo[Cys-D-Val-Gln-D-Phe-Arg-Trp-Cys]-NH2; (SEQ ID NO: 11), or Ac-Arg-cyclo[Cys-D-Val-His(1-Me)-D-Phe-Arg-Trp-Cys]-NH2 (SEQ ID NO: 58), and include polypeptides represented by any of them or pharmaceutically acceptable salts thereof.

[0118] In a further embodiment, the polypeptide of the present invention is represented by the following structural formula: Ac-TzAla-cyclo[Cys-Ala-Gln-D-Phe-Arg-Trp-Cys]-NH2; (SEQ ID NO: 59), or Ac-Glu-cyclo[Cys-Ala-His-D-Phe-Arg-Trp-Cys]-NH2 (SEQ ID NO: 60), and include polypeptides represented by any of them or pharmaceutically acceptable salts thereof.

[0119] In yet another embodiment, the polypeptide of the present invention is represented by the following structural formula: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Arg-Trp-Cys]-NH2; (SEQ ID NO: 37);​​ Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 8), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 7), or a polypeptide represented by any of them, or a pharmaceutically acceptable salt thereof.

[0120] In a further embodiment, the polypeptide of the present invention has the following structural formula: Ac-Arg-cyclo[Cys-D-Leu-His-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 61) Ac-Arg-cyclo[Cys-D-Ile-His-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 62) Ac-Arg-cyclo[Cys-D-Tle-His-D-Phe-Arg-Trp- Cys]-NH2; (SEQ ID NO: 63), or Ac-Arg-cyclo[Cys-D-Val-His-D-Phe-Arg-Trp- Cys]-NH2 (SEQ ID NO: 10), or a polypeptide represented by any of them, or a pharmaceutically acceptable salt thereof.

[0121] In a further embodiment, the polypeptide of the present invention has the following structural formula: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-2-Nal- Arg-Trp-Cys]-NH2; (SEQ ID NO: 64) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-2-Nal-Arg-Tr p-Cys]-NH2; (SEQ ID NO: 65), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-2-Nal-Arg-Tr A polypeptide represented by any of p-Cys]-NH2 (SEQ ID NO: 66), or includes a pharmaceutically acceptable salt thereof.

[0122] In a further embodiment, the polypeptide of the present invention has the following structural formula: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Ar g-Trp-Cys]-OH; (SEQ ID NO: 67) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- Cys]-OH; (SEQ ID NO: 68), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-OH (SEQ ID NO: 69), or is a pharmaceutically acceptable salt thereof.

[0123] [Examples] (Peptide synthesis) The peptides of the present invention were prepared by conventional solid-phase peptide synthesis. The peptide chain was initiated from a C-terminal amino acid derivative coupled to a suitably selected solid support resin known to be suitable for peptide synthesis and extended in a stepwise manner. For peptide synthesis with an amide functional group at the C-terminus, Rink amide MBHA resin was used as the solid support. For peptide synthesis with a free carboxyl functional group at the C-terminus, a resin (e.g., 2-chlorotrityl chloride resin, Wang resin, or Merrifield resin ) may be utilized to create an ester bond with an Fmoc-amino acid. Most of these Fmoc-amino acid-resin types with linked esters are commercially available from various sources and are generally used when feasible. Most of these Fmoc-amino acid-resin types with linked esters are commercially available from various sources and are generally used when feasible.

[0124] (Synthesis of Disulfide-Cyclized Peptides) The linear derivative of the disulfide cyclic peptide amide was constructed using solid-phase peptide synthesizer and Fmoc chemistry. Fmoc-Rink amide resin was placed in the reaction vessel and swollen with NMP. It was treated with 20% piperidine in NMP for 15 minutes and then washed three times with NMP. The resin was tested using the positive Kaiser test (Kaiser, E., Colescot, R.L., B ossinge, C.D. & Cook, P.I, Anal. Biochem., 199 0, 34:595-598). It was resuspended in NMP and mixed with the required first C-terminal Fmoc-amino acid derivative and HOBt. The coupling reaction was initiated by adding HBTU reagent and DIEA. After mixing for 2-3 hours, the completion of the coupling was confirmed by the negative Kaiser test from a small aliquot of the resin removed from the reaction mixture. Then the resin was washed three times with NMP. Subsequently, the Fmoc group was removed as described and the whole cycle was repeated as above using the second C-terminal Fmoc-amino acid derivative. The same reaction cycle was sequentially repeated using each incoming amino acid. The chloranil color test (Vojkovsky, T. Pept. Res., 1995, 8:236-237) was used instead of the Kaiser test for the positive test of Fmoc deprotection from proline residues in the peptide sequence and for testing the completion of amino acid coupling to proline (negative chloranil test). For peptides having an acetyl group at the N-terminus, the Fmoc-deprotected peptide resin was treated with acetic anhydride and pyridine for 10 minutes. The resin that was negative in the Kaiser test was N MP. ​​It was washed with MP and dichloromethane and dried under reduced pressure. The Fmoc-amino acid derivative was used for the synthesis of these peptides. The trifunctional amino acid derivatives used were Fmoc-Cys(Trt)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Arg(Pbf)-OH, Fmoc-His(Trt)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-hCys(Trt)-OH, Fmoc-Pen(Trt)-OH, Fmoc-Tyr(But)-OH, Fmoc-His(1-Me)-OH, Fmoc-His(3-Me)-OH and Fmoc-Glu(OBut)-OH. To cleave the peptide from the resin and deprotect the side-chain functional groups, the peptide resin was taken up in 2% TIS / 5% water / 5% (w / v) DTT / 88% TFA. This solution was mixed for 3.5 hours and then filtered. The filtrate was mixed with cold anhydrous ethyl ether. The precipitate was collected by centrifugation. The solvent was decanted and the peptide pellet was resuspended in fresh ether. The ether work was repeated two or more times. The peptide was dried under reduced pressure. The crude linear peptide product was diluted in 5% acetic acid to a concentration of 2 mg / mL and 0.5 M iodine / methanol was added dropwise with vigorous stirring until a persistent pale yellow color of the solution was achieved. This solution was stirred for an additional 10 minutes. Excess iodine was then quenched by adding 1 M sodium thiosulfate while mixing until the mixture became colorless. The cyclized peptide solution was lyophilized and the crude powder was purified by preparative HPLC using a reversed-phase C-18 column. The fractions of the purified product were collected and lyophilized. Cys(Trt)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Arg(Pb f)-OH, Fmoc-His(Trt)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-hCys(Trt)-OH, Fmoc- Pen(Trt)-OH, Fmoc-Tyr(But)-OH, Fmoc-His(1- Me)-OH, Fmoc-His(3-Me)-OH and Fmoc-Glu(OBut )-OH.

[0125] To cleave the peptide from the resin and deprotect the side-chain functional groups, the peptide resin was taken up in 2% TIS / 5% water / 5% (w / v) DTT / 88% TFA. This solution was mixed for 3.5 hours and then filtered. The filtrate was mixed with cold anhydrous ethyl ether. The precipitate was collected by centrifugation. The solvent was decanted and the peptide pellet was resuspended in fresh ether. The ether work was repeated two or more times. The peptide was dried under reduced pressure. The crude linear peptide product was diluted in 5% acetic acid to a concentration of 2 mg / mL and 0.5 M iodine / methanol was added dropwise with vigorous stirring until a persistent pale yellow color of the solution was achieved. This solution was stirred for an additional 10 minutes. Then, excess iodine was quenched by adding 1 M sodium thiosulfate while mixing until the mixture became colorless. The cyclized peptide solution was lyophilized and the crude powder was purified by preparative HPLC using a reversed-phase C-18 column. The fractions of the purified product were collected and lyophilized. Using electrospray ionization technology, peptides were analyzed and identified by mass spectrometry , and the mass was corrected.

[0126] (Synthesis of lactam-cyclized peptides) Cyclic lactam peptides were also synthesized by standard solid-phase peptide synthesis methods. For peptides having Dpr at the C-terminus, Fmoc-Dpr(Mtt)-BHA resin was transferred to a solid-phase peptide synthesis reactor. The Fmoc group was removed as described above, and the next Fmoc-protected amino acid, e.g., Fmoc-Trp(Boc)-OH, was coupled to the resin by standard coupling procedures . The Fmoc protecting group was removed, and coupling and deprotection were repeated until the amino acid sequence was complete, adding the remaining amino acids individually to the correct sequence. For glutamic acid, Fmoc-Glu(OPip) coupling was used . Then, the fully aligned peptides were acetylated at the N-terminus as described for the disulfide series of peptides. Next, the orthogonally protected side chains were removed. For example, a peptide resin having an orthogonally protected Glu side chain as a 2-phenylisopropyl (O Pip) ester or having Dpr as 4-methyltrityl (Mtt) was cleaved by treatment with 1% TFA in dichloromethane . The deprotected peptide resin was suspended in NMP and treated with HBTU / DIPEA. After cyclization (negative Kaiser test), the peptide-resin was washed with DCM and dried . The cyclic peptide was cleaved from the resin with trifluoro acetic acid (TFA) in the presence of water and 1,2-ethanedithiol (EDT), along with the remaining protecting groups. Cold anhydrous ether . After cyclization (negative Kaiser test), the peptide-resin was washed with DCM and dried . The cyclic peptide was cleaved from the resin with trifluoro acetic acid (TFA) in the presence of water and 1,2-ethanedithiol (EDT), along with the remaining protecting groups. Cold anhydrous ether was added and the product was collected by precipitation and by centrifugation. Final purification was by reverse phase C -18 column and by reverse phase HPLC. The purified peptide was lyophilized and collected, and the mass was analyzed by mass spectrometry using electrospray methodology.

[0127] Examples of the compounds of the present invention are given in Table 2. Table 2: Examples of the compounds of the present invention

Table 3

[0128] Radioactive ligand binding assay A receptor binding assay to determine the binding constant (K d ), or an inhibition concentration (IC ) for replacing a radiolabeled ligand from the receptor of the cyclic peptide of the present invention, may be carried out by any means known in 50 the art. For example, the preparation of cell membranes for the binding assay is prepared from CHO-K1 cells transfected to stably express hMC receptor subtypes 1, 3,

[0129] 4 or 5. The competitive inhibition of [125I](Tyr2)-(Nle4-D-Phe7)-α-MSH ([125I -NDP-a-MSH binding is carried out in a polypropylene 96 well plate. Briefly, the cell membranes prepared as described above (1 - 10 μg of protein / well) are used, increasing the concentration of the test compound up to 0.1 - 0.3 nM of [125I]-NDP-α-MSH, while including 0.2% BSA, 5 mM MgCl2, 1 mM CaCl2 and 0.1 mg . while increasing the concentration of the test compound to [125I]-NDP-α-MSH of 0.1 - 0.3 nM while including 0.2% BSA, 5 mM MgCl2, 1 mM CaCl2 and 0.1 mg in 50 mM Tris-HCl containing bacitracin at about 120 minutes at pH 7.4 and 37 °C. The bound [125I]-NDP-α-MSH ligand was separated from free [125I]-NDP-α-MSH by filtration through GF / C glass fiber plates (Unifilter®, Meriden, CT, USA) pre-soaked in 0.1% (w / v) polyethyleneimine (PEI). The fil ters were washed three times with 50 mM Tris-HCl at pH 7.4 at a temperature of about 0-4 °C and then assayed for radioactivity. The binding data were analyzed by computer-assisted non-linear regression analysis.

[0130] (Stimulation assay of cyclic AMP) The functional assay for determining the agonist or antagonist status of the cyclic peptides of the present invention may be performed by any means known in the art. (Electrochemiluminescence (ECL) assay) The stimulation of intracellular cyclic AMP (cAMP) levels by the peptide was determined in a dose-dependent manner by electrochemiluminescence (ECL ) assay (Meso Scale Discovery, Gaithersburg, MD, USA; hereinafter referred to as "MSD" in this specification) . Briefly, CHO-K1 cells stably expressing the hMC receptor subtype were suspended in RPM I1640® assay buffer (RPMI 1640 buffer containing 0.5 mM IBMX and 0.2% protein cocktail (MSD blocker A)). Approximately 7,000 cells stably expressing the hMC receptor subtype 1, 3, 4 or 5 were suspended in RPMI 1640 buffer containing 0.5 mM IBMX and 0.2% protein cocktail (MSD blocker A)). were suspended in RPMI 1640 buffer containing 0.5 mM IBMX and 0.2% Transgenic CHO-K1 cells in the wells are dispensed into a 384-well Multi-Array Plate (MSD) containing integrated carbon electrodes and coated with anti-cAMP antibody. Increasing concentrations of the test compound are added and the cells are incubated at 37 °C for about 40 minutes. A cell lysis buffer (HEPES-buffered saline solution containing 0.2% protein cocktail, 2.5 nM ruthenium-labeled cAMP (MSD), MgCl2 and Triton X-100 (registered trademark) at pH 7.3) is added and the cells are incubated at room temperature for about 90 minutes. At the end of the second incubation time, a reading buffer (Tris-buffered solution containing ECL co-reagent and Triton X-100 at pH 7.8) is added and the cAMP levels in the cell lysate are immediately determined by ECL detection using a Sector Imager 6000 reader (registered trademark) (MSD). The data are analyzed by computer-assisted non-linear regression analysis (XL Fit; IDBS) and reported as either EC50 value. The EC50 represents the concentration of the agonist compound required to obtain 50% of the maximum response (e.g., 50% of the maximum level of cAMP as determined using the assay described above). Cells are transfected with human MC4-R in 96-well plates and grown to confluent state (seeded at approximately 250,000 cells / well). Three of these cells are treated with 0.2 mM isobutylmethylxanthine (IBMX) and peptides at stepwise concentrations or peptides in the presence of 20 nM NDP-MSH. 20 nM NDP-M TM ruthenium-labeled cAMP (MSD) is added, and the cells are incubated at room temperature for about 90 minutes. At the end of the second incubation time, a reading buffer (Tris-buffered solution containing ECL co-reagent and Triton X-100 at pH 7.8) is added and the cAMP levels in the cell lysate are immediately determined by ECL detection using a Sector Imager 6000 reader (registered trademark) (MSD). The data are analyzed by computer-assisted non-linear regression analysis (XL Fit; IDBS) and reported as either EC50 value. The EC50 represents the concentration of the agonist compound required to obtain 50% of the maximum response (e.g., 50% of the maximum level of cAMP as determined using the assay described above). HEPES-buffered saline solution containing 0.2% protein cocktail, 2.5 nM ruthenium-labeled cAMP (MSD), MgCl2 and Triton X-100 (registered trademark) at pH 7.3) is added, and the cells are incubated at room temperature for about 90 minutes. At the end of the second incubation time, a reading buffer (Tris-buffered solution containing ECL co-reagent and Triton X-100 at pH 7.8) is added and the cAMP levels in the cell lysate are immediately determined by ECL detection using a Sector Imager 6000 reader (registered trademark) (MSD). The data are analyzed by computer-assisted non-linear regression analysis (XL Fit; IDBS) and reported as either EC50 value. The EC50 represents the concentration of the agonist compound required to obtain 50% of the maximum response (e.g., 50% of the maximum level of cAMP as determined using the assay described above). A reading buffer (Tris-buffered solution containing ECL co-reagent and Triton X-100 at pH 7.8) is added, and the cAMP levels in the cell lysate are immediately determined by ECL detection using a Sector Imager 6000 reader (registered trademark) (MSD). The data are analyzed by computer-assisted non-linear regression analysis (XL Fit; IDBS) and reported as either EC50 value. The EC50 represents the concentration of the agonist compound required to obtain 50% of the maximum response (e.g., 50% of the maximum level of cAMP as determined using the assay described above). Sector Imager 6000 reader (registered trademark) (MSD). The data are analyzed by computer-assisted non-linear regression analysis (XL Fit; IDBS) and reported as either EC50 value. The EC50 represents the concentration of the agonist compound required to obtain 50% of the maximum response (e.g., 50% of the maximum level of cAMP as determined using the assay described above). The data are analyzed by computer-assisted non-linear regression analysis (XL Fit; IDBS) and reported as either EC50 value. The EC50 represents the concentration of the agonist compound required to obtain 50% of the maximum response (e.g., 50% of the maximum level of cAMP as determined using the assay described above). 50% of the maximum response (e.g., 50% of the maximum level of cAMP as determined using the assay described above).

[0131] (cAMP measurement assay) Cells transfected with human MC4-R in 96-well plates are grown to confluent state (seeded at approximately 250,000 cells / well). Three of these cells are treated with 0.2 mM isobutylmethylxanthine (IBMX) and peptides at stepwise concentrations or peptides in the presence of 20 nM NDP-MSH. 20 nM NDP-M ​​​​​​​​Cells that are similarly processed with SH alone function as a positive control in a volume of 200 μL. A buffer blank that functions as a negative control is also included. After incubation at 37 °C for 1 hour, 50 μL of cell lysis buffer is added to lyse the cells. The total cAMP accumulated in this 250 μL incubation medium is quantified using a commercially available low pH cAMP assay kit (Amersham Biosciences) according to the procedure specified by the kit supplier. A peptide that shows cAMP accumulation in the same range or a greater range as α-MSH as a positive control is considered an agonist. The agonist data is plotted, the curve is fitted, and the EC50 value is determined. A peptide that shows accumulation in the same range as the negative control (buffer blank in the absence of α-MSH) is not effective at the test concentration. A peptide that shows attenuated accumulation is considered an antagonist if inhibition is present in cAMP when α-MSH is also present in the assay. A similar assay may be performed using hMC-1R cells, hMC-3R cells, and hMC-5R cells. Included is also a buffer blank that functions as a negative control. After incubation at 37 °C for 1 hour, 50 μL of cell lysis buffer is added to lyse the cells. After incubation at 37 °C for 1 hour, 50 μL of cell lysis buffer is added to lyse the cells. The total cAMP accumulated in this 250 μL incubation medium is quantified using a commercially available low pH cAMP assay kit (Amersham Biosciences) according to the procedure specified by the kit supplier. The total cAMP accumulated in this 250 μL incubation medium is quantified using a commercially available low pH cAMP assay kit (Amersham Biosciences) according to the procedure specified by the kit supplier. nces) according to the procedure specified by the kit supplier. A peptide that shows cAMP accumulation in the same range or a greater range as α-MSH as a positive control is considered an agonist. A peptide that shows cAMP accumulation in the same range or a greater range as α-MSH as a positive control is considered an agonist. The agonist data is plotted, the curve is fitted, and the EC50 value is determined. A peptide that shows accumulation in the same range as the negative control (buffer blank in the absence of α-MSH) is not effective at the test concentration. The agonist data is plotted, the curve is fitted, and the EC50 value is determined. A peptide that shows accumulation in the same range as the negative control (buffer blank in the absence of α-MSH) is not effective at the test concentration. A peptide that shows accumulation in the same range as the negative control (buffer blank in the absence of α-MSH) is not effective at the test concentration. A peptide that shows attenuated accumulation is considered an antagonist if inhibition is present in cAMP when α-MSH is also present in the assay. A peptide that shows attenuated accumulation is considered an antagonist if inhibition is present in cAMP when α-MSH is also present in the assay. A similar assay may be performed using hMC-1R cells, hMC-3R cells, and hMC-5R cells.

[0132] (Measurement of cAMP Accumulation Using the β-Galactosidase (β-Gal) Reporter System) (Measurement of cAMP Accumulation Using the β-Galactosidase (β-Gal) Reporter System) A chemiluminescence reading system using an enzyme fragment complementation (EFC) system that uses β-galactosidase (β-Gal) as a functional reporter system was used. This assay system for various melanocortin receptor systems is commercially available (cAMP Hunter GPCR assay system, Discoverx Corp, Fremont, A chemiluminescence reading system using an enzyme fragment complementation (EFC) system that uses β-galactosidase (β-Gal) as a functional reporter system was used. This assay system for various melanocortin receptor systems is commercially available (cAMP Hunter GPCR assay system, Discoverx Corp, Fremont, A chemiluminescence reading system using an enzyme fragment complementation (EFC) system that uses β-galactosidase (β-Gal) as a functional reporter system was used. This assay system for various melanocortin receptor systems is commercially available (cAMP Hunter GPCR assay system, Discoverx Corp, Fremont, A chemiluminescence reading system using an enzyme fragment complementation (EFC) system that uses β-galactosidase (β-Gal) as a functional reporter system was used. This assay system for various melanocortin receptor systems is commercially available (cAMP Hunter GPCR assay system, Discoverx Corp, Fremont, (CA). This assay utilizes the β-Gal enzyme that is divided into two complementary parts, EA for the enzyme acceptor and ED for the enzyme donor. In this assay, the ED portion fused to cAMP is made to compete with the binding of cAMP produced by the cells to an antibody specific for cAMP. Then, EA is added to create active β-Gal containing unbound ED-cAMP. This active enzyme then converts a chemiluminescent substrate to generate an output signal, which is recorded with a standard microplate reader. Briefly, inoculate 10,000 cells / well overnight, and then incubate each well (incubating the cells with 10 μl of assay buffer) in cell assay buffer (5 μL) with test compounds and cAMP antibody reagent (5 μL) at four-fold serial dilutions at 37 °C for 30 minutes. Then, add cell lysis buffer (20 μL) containing the enzyme fragment coupled with ED-cAMP and a reporter substance (Emerald II-Galacton Star, 5:1), and incubate at room temperature for 60 minutes. Next, add 20 μL of EA β-Gal fragment reagent. After further incubation at room temperature for 120 minutes, measure the chemiluminescence with a plate reader (Envision), and use the data to calculate the EC50 value for the test peptide. The ED part fused to cAMP is made to compete with the binding of cAMP produced by the cells to an antibody specific for cAMP. Then, EA is added to create active β-Gal containing unbound ED-cAMP. This active enzyme then converts a chemiluminescent substrate to generate an output signal, which is recorded with a standard microplate reader. This active enzyme then converts a chemiluminescent substrate to generate an output signal, which is recorded with a standard microplate reader. This active enzyme then converts a chemiluminescent substrate to generate an output signal, which is recorded with a standard microplate reader.

[0133] Briefly, inoculate 10,000 cells / well overnight, and then incubate each well (incubating the cells with 10 μl of assay buffer) in cell assay buffer (5 μL) with test compounds and cAMP antibody reagent (5 μL) at four-fold serial dilutions at 37 °C for 30 minutes. Then, add cell lysis buffer (20 μL) containing the enzyme fragment coupled with ED-cAMP and a reporter substance (Emerald II-Galacton Star, 5:1), and incubate at room temperature for 60 minutes. This active enzyme then converts a chemiluminescent substrate to generate an output signal, which is recorded with a standard microplate reader. The ED part fused to cAMP is made to compete with the binding of cAMP produced by the cells to an antibody specific for cAMP. Then, EA is added to create active β-Gal containing unbound ED-cAMP. This active enzyme then converts a chemiluminescent substrate to generate an output signal, which is recorded with a standard microplate reader. Then, add cell lysis buffer (20 μL) containing the enzyme fragment coupled with ED-cAMP and a reporter substance (Emerald II-Galacton Star, 5:1), and incubate at room temperature for 60 minutes. Next, add 20 μL of EA β-Gal fragment reagent. After further incubation at room temperature for 120 minutes, measure the chemiluminescence with a plate reader (Envision), and use the data to calculate the EC50 value for the test peptide. Next, add 20 μL of EA β-Gal fragment reagent. After further incubation at room temperature for 120 minutes, measure the chemiluminescence with a plate reader (Envision), and use the data to calculate the EC50 value for the test peptide.

[0134] The results are shown in Table 3.

[0135] Table 3: EC50 (nM) values of the compounds of the examples of the present invention [Table 4] TIFF2025102830000024.tif168135

[0136] The present invention has been specifically shown and described with reference to the embodiments of the examples, but various changes in form and detail will be understood by those skilled in the art to be made without departing from the scope of the present invention encompassed by the appended claims. ​

Claims

1. An isolated polypeptide of the following structural formula (I): 【Chemical 1】 or a pharmaceutically acceptable salt thereof, wherein wherein R １ is -H or C1-C6 acyl; R ２ is - NR ３ R ４ or - OR ５ and R ３ R ４ and R ５ are each independent is H or C1-C6 alkyl; A １ is Arg, Lys, Orn, His, Nle, Phe, Val, Leu, Trp is an amino acid residue selected from Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzAla; or A １ is optionally substituted C1-C12 alkyl, optionally substituted C6-C is aryl having 18 carbon atoms, optionally substituted C5-C18 heteroaryl, aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; is aryl having 18 carbon atoms, optionally substituted C5-C18 heteroaryl, aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; is aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; is aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; or A １ does not exist; or A ２ and A ８ are each independently Cys, hCys, Pen, Asp, Glu, An amino acid residue selected from Lys, Orn, Dbu, or Dpr, and A ２ and A ８ is selected to be a pair such that a covalent bond can be formed between the respective side chains; A ３ is Ala, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr is an amino acid residue selected from Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib; or A ３ does not exist or A ３ is residue Y, and Y has the following structural formula 【Chemical Formula 2】 is an amino acid selected from the amino acids represented by R １１ and R １２ are each independently H, -CH ３ , phenyl or benzyl wherein R ２１ 、 R ２２ 、 R ２３ and R ２４ are each independently H, -CH ３ , -CF ３ , phenyl, benzyl, F, Cl, Br, I, -OCH ３ , or -OH; R ３１ 、R ３２ 、R ３３ 、R ３４ 、R ４１ 、R ４２ and R ４３ are each independently , H, -CH ３ , -CF ３ , phenyl, benzyl, F, Cl, Br, I, -OCH ３ , or is -OH; A ４ is absent or is ATC, Ala, QAla, Aib, Sar, Ser is an amino acid selected from Thr, Pro, Hyp, Asn, Gln, optionally substituted His, Trp, Tyr, Lys, Arg, sChp, or residue X, where X is is an amino acid selected from the amino acids represented by the following structural formula wherein [Chemical 3] 【Chem.】 is an amino acid selected from the amino acids represented by the following structural formula wherein R ５１ and R ５２ each independently represents H, -CH ３ , phenyl, or benzyl is; R ６１ , R ６２ , R ６３ and R ６４ are each independently H, -CH ３ , -CF ３ , phenyl, benzyl, F, Cl, Br, I, -OCH ３ , or -OH; R ７１ 、R ７２ 、R ７３ 、R ７４ 、R ８１ 、R ８２ and R ８３ are each independently , H, -CH ３ , -CF ３ , phenyl, benzyl, F, Cl, Br, I, -OCH ３ , or is -OH; A ５ is optionally substituted Phe, optionally substituted 1-Nal, or substitution is optionally substituted 2-Nal; A ６ is Arg; A ７ is Trp, any amino acid residue is in either the L-configuration or the D-configuration, provided that (1) A ３ and A ４ are both not non-existent; (2) A ４ When A is an amino acid, ３ A is neither Aib nor Gly; (3) A ４ is His, and A ５ is D-Phe or 2-Nal, then A ３ is it is neither a D-amino acid nor L-Ala; (4) A ２ and A ８ are each selected from Cys, hCys or Pen together When (a) A ４ is absent, A ３ is not L-His; (b) When A does not exist, ３ A is not L-His; ４ instead, (c) A ４ When A is His, ３ A is not Glu, Leu, or Lys, a polypeptide.

2. R １ is -H or C1-C6 acyl; R ２ is -NR ３ R ４ or -OR ５ and R ３ R ４ and R ５ are each independently standing, is H or C1-C6 alkyl; A １ Does not exist; or A １ is Arg, Lys, Orn, His, Nle, Phe, Val, Leu, Trp is an amino acid residue selected from Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzAla; or A １ is optionally substituted C1-C12 alkyl, optionally substituted C6-C is aryl having 18 carbon atoms, optionally substituted C5-C18 heteroaryl, aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; is aryl having 18 carbon atoms, optionally substituted C5-C18 heteroaryl, aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; is aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; is aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; is a moiety selected from aralkyl wherein the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl wherein the heteroaryl moiety is optionally substituted C5-C18 heteroaryl and the alkyl moiety is optionally substituted C1-C12 alkyl; A ２ and A ８ are each independently Cys, hCys, Pen, Asp, Glu, An amino acid residue selected from Lys, Orn, Dbu, or Dpr, and A ２ and A ８ is selected in pairs so as to be able to form a covalent bond between the respective side chains; A ３ either does not exist or is Ala, Tle, Val, Leu, Ile, Cha, is an amino acid residue selected from Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib; is an amino acid residue selected from Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib; A ４ is absent or is ATC, Ala, Qala, Aib, Sar, Ser , Thr, Pro, Hyp, Asn, Gln, optionally substituted His, Trp, Tyr, Lys, Arg, sChp, or residue X, where X is an amino acid residue selected from the following amino acids represented by the structural formula , or 2-Nal which may be substituted; 【Chemical Formula 4】 wherein any amino acid residue is in either the L-configuration or the D-configuration, the isolated polypeptide according to claim 1 A ５ is optionally substituted Phe, optionally substituted 1-Nal, or substitution wherein X is an amino acid selected from the amino acids represented by the following structural formula A ６ is Arg; A ７ is Trp, wherein any amino acid residue is in either the L-configuration or the D-configuration, the isolated polypeptide according to claim 1 . **Claim 3** A ３ The polypeptide according to claim 1 or 2, wherein A is a D-amino acid. **Claim 4** A ４ The polypeptide according to claim 1 or 2, wherein A is an L-amino acid. **Claim 5** A ４ The polypeptide according to claim 1 or 2, wherein A does not exist. **Claim 6** A ５ The polypeptide according to any one of claims 1 to 5, wherein A is D-Phe which may be substituted 。 **Claim 7** A ５ is F, Cl, Br, I, -CH ３ , -OH, -CN, amine, -NO ２ or -OCH ３ substituted with any of five aromatic carbons having a substituent selected from The polypeptide according to any one of claims 1 to 6, which may also be... **Claim 8** A ５ is Phe, Phe(2'-F), Phe(2'-Cl), Phe(2'-Br) , Phe(2'-I), Phe(2'-CN), Phe(2'-CH ３ ), Phe(2' -OCH ３ ), Phe(2'-CF ３ ), Phe(2'-NO ２ ), Phe(3'-F) , Phe(3'-Cl), Phe(3'-Br), Phe(3'-I), Phe(3'- CN), Phe(3'-CH ３ ), Phe(3'-OCH ３ ), Phe(3'-CF ３ ), , Phe(3'-NO ２ ), Phe(4'-F), Phe(4'-Cl), Phe(4' -Br), Phe(4'-I), Phe(4'-CN), Phe(4'-CH ３ ), Ph e(4'-OCH ３ ), Phe(4'-CF ３ ), Phe(4'-NO ２ ), Phe(4 '-t-Bu), Phe(2',4'-diF), Phe(2',4'-diCl), Phe (2',4'-diBr), Phe(2',4'-diI), Phe(2',4'-di-CN ), Phe(2',4'-di-CH ３ ), Phe(2',4'-di-OCH ３ ), Phe (3',4'-diF), Phe(3',4'-diCl), Phe(3',4'-diBr) , Phe(3',4'-diI), Phe(3',4'-di-CN), Phe(3',4' -Di-CH ３ ), Phe(3',4'-Di-OCH ３ ), Phe(3',5'-DiF), Phe(3',5'-diCl), Phe(3',5'-diBr), Phe(3',5'- D-I), Phe(3',5'-di-CN), Phe(3',5'-diCH ３ ), Phe( 3',5'-Di-OCH ３ ), or selected from Phe(3',4',5'-TrF) which is a D-amino acid residue, the polypeptide according to any one of claims 1 to 6. **Claim 9** A ４ is F, Cl, Br, I, -CH ３ , -OH, -CN, amine, -NO ２ or -OCH ３ His which is a substituent selected from the group consisting of, and may be substituted at any substitution position The polypeptide according to any one of claims 1 to 4 or 6 to 8, which is... **Claim 10** The polypeptide according to claim 1 or 2, represented by any one of the following structural formulas , or a pharmaceutically acceptable salt thereof. 【Chemical Formula 5】 【Chem.】 **Claim 11** The polypeptide according to claim 1 or 2, represented by any one of the following structural formulas , or a pharmaceutically acceptable salt thereof. 【Chemical Formula 6】 **Claim 12** The polypeptide according to claim 1 or 2, represented by any one of the following structural formulas , or a pharmaceutically acceptable salt thereof. 【Chemical Formula 7】 **Claim 13** A ４ is Atc, Ala, QAla, Aib, Sar, Ser, Thr, Pro, Hy p, Asn, Gln, substituted His, Trp, Tyr, Lys, Arg, sChp, or an amino acid residue selected therefrom, or residue X, the polypeptide according to claim 1 or 2 . **Claim 14** The following structural formula: Ac-Arg-cyclo[Cys-D-Ala-His(3-Me)-D-Phe-Arg- g-Trp-Cys]-NH ２ ; (SEQ ID NO: 36) Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Arg- g-Trp-Cys]-NH ２ ; (SEQ ID NO: 37) Ac-Arg-cyclo[Cys-D-Ala-Trp-D-Phe-Arg-Trp- [Cys]-NH ２ ; (SEQ ID NO: 9) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- [Cys]-NH ２ ; (SEQ ID NO: 8) Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-NH ２ ; (SEQ ID NO: 7) Ac-Arg-cyclo[Cys-D-Ala-Arg-D-Phe-Arg-Trp- [Cys]-NH ２ ; (SEQ ID NO: 38) Ac-Arg-cyclo[Cys-D-Ala-Tyr-D-Phe-Arg-Trp- Cys]-NH ２ ; (SEQ ID NO: 39) Ac-Arg-cyclo[Cys-D-Ala-D-Pro-D-Phe-Arg-Tr p-Cys]-NH ２ ; (SEQ ID NO: 40) Ac-Arg-cyclo[Cys-D-Ala-Pro-D-Phe-Arg-Trp- [Cys]-NH ２ ; (SEQ ID NO: 2) Ac-Arg-cyclo[Cys-D-Ala-Pro-D-Phe(p-F)-Arg -Trp-Cys]-NH ２ ; (SEQ ID NO: 4) Ac-Arg-cyclo[Cys-D-Ala-Atc-D-Phe-Arg-Trp- Cys]-NH ２ ; (SEQ ID NO: 41) Ac-Arg-cyclo[Cys-D-Ala-QAla-D-Phe-Arg-Trp -Cys]-NH ２ ; (SEQ ID NO: 42) Ac-Arg-cyclo[Cys-D-Ala-sChp-D-Phe-Arg-Trp -Cys]-NH ２ ; (SEQ ID NO: 43), or Ac-Arg-cyclo[Cys-D-Ala-X-D-Phe-Arg-Trp-Cy s]-NH ２ represented by any one of (SEQ ID NO: 44), or its pharmaceutically The polypeptide according to claim 13, which is an acceptable salt.

15. The following structural formula: Ac-Arg-cyclo[hCys-Ala-D-Phe-Arg-Trp-Cys]- NH ２ ; (SEQ ID NO: 15) Ac-Arg-cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Cys -NH ２ ; (SEQ ID NO: 14) Ac-Arg-cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Pen -NH ２ ; (SEQ ID NO: 45) Ac-Arg-cyclo[Glu-D-Ala-D-Phe-Arg-Trp-Dpr -NH ２ ; (SEQ ID NO: 26) Ac-Arg-cyclo[Glu-Ala-D-Phe-Arg-Trp-Dpr]-N H ２ ; (SEQ ID NO: 27) Ac-Arg-cyclo[hCys-Aib-D-Phe-Arg-Trp-Cys]- NH ２ ; (SEQ ID NO: 46) Ac-Arg-cyclo[hCys-Sar-D-Phe-Arg-Trp-Cys]- NH ２ ; (SEQ ID NO: 47) Ac-Arg-cyclo[hCys-Val-D-Phe-Arg-Trp-Cys]- NH ２ ; (SEQ ID NO: 48) Ac-Arg-cyclo[hCys-D-Val-D-Phe-Arg-Trp-Cys -NH ２ ; (SEQ ID NO: 49) Ac-Arg-cyclo[hCys-Gln-D-Phe-Arg-Trp-Cys]- NH ２ ; (SEQ ID NO: 50) Ac-Arg-cyclo[hCys-D-Gln-D-Phe-Arg-Trp-Cys -NH ２ ; (SEQ ID NO: 51) Ac-Arg-cyclo[hCys-Ala-D-Phe-Arg-Trp-Pen]- NH ２ ; (SEQ ID NO: 52) Ac-Arg-cyclo[D-Pen-D-Ala-D-Phe-Arg-Trp-hC ys]-NH ２ ; (SEQ ID NO: 53) Ac-Arg-cyclo[Cys-D-Ala-D-Phe-Arg-Trp-hCys -NH ２ ; (SEQ ID NO: 17) Ac-Arg-cyclo[Pen-D-Ala-D-Phe-Arg-Trp-hCys - NH ２ ; (SEQ ID NO: 54) Ac-Arg-cyclo[D-hCys-D-Ala-D-Phe-Arg-Trp-C ys]-NH ２ ; (SEQ ID NO: 55) Ac-Arg-cyclo[hCys-Pro-D-Phe-Arg-Trp-Cys]- NH ２ ; (SEQ ID NO: 20), or Ac-Arg-cyclo[hCys-D-Pro-D-Phe-Arg-Trp-Cys -NH ２ represented by any one of (SEQ ID NO: 56), or pharmaceutically acceptable The polypeptide according to claim 1 or 2, which is a pharmaceutically acceptable salt thereof.

16. A ３ is Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Lys An amino acid residue selected from Arg, His, Phe, Gln, Sar, Gly, Asn, or Aib; ; A ４ is Atc, Ala, QAla, Aib, Sar, Ser, Thr, Pro, Hy p, an amino acid residue selected from Asn, Gln, substituted His, Trp, Tyr, Lys, Arg, sChp, or residue X, the polypeptide according to claim 1 or 2.

17. The following structural formula: Ac-Arg-cyclo[Cys-Val-Gln-D-Phe-Arg-Trp-Cy (SEQ ID NO: 57) Ac-Arg-cyclo[Cys-D-Val-Gln-D-Phe-Arg-Trp- s]-NH ２ ; (SEQ ID NO: 11), or Ac-Arg-cyclo[Cys-D-Val-His(1-Me)-D-Phe-Ar [Cys]-NH ２ ; The polypeptide according to claim 16, which is a pharmaceutically acceptable salt thereof.

18. g-Trp-Cys]-NH ２ represented by any of (SEQ ID NO: 58), or The following structural formula: Ac-TzAla-cyclo[Cys-Ala-Gln-D-Phe-Arg-Trp- Ac-Glu-cyclo[Cys-AIa-His-D-Phe-Arg-Trp-Cy The polypeptide according to claim 1 or 2, which is a pharmaceutically acceptable salt thereof. [Cys]-NH ２ ; (SEQ ID NO:59), or

19. s]-NH ２ represented by any of (SEQ ID NO: 60), or its pharmaceutically acceptable The following structural formula: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Ar (SEQ ID NO: 37) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- g-Trp-Cys]-NH ２ ; (SEQ ID NO: 8), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- [Cys]-NH ２ ; ​ ​ [Cys]-NH ２ represented by any of (SEQ ID NO: 7), or its pharmaceutically The polypeptide according to claim 1 or 2, which is an acceptable salt.

20. The following structural formula: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-2-Nal- Arg-Trp-Cys]-NH ２ ; (SEQ ID NO: 64) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-2-Nal-Arg-Tr [p-Cys]-NH ２ ; (SEQ ID NO: 65), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-2-Nal-Arg-Tr p-Cys]-NH ２ represented by any of (SEQ ID NO: 66), or a medicament thereof The polypeptide according to claim 1 or 2, which is a pharmaceutically acceptable salt.

21. The following structural formula: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Arg- Trp-Cys]-OH; (SEQ ID NO: 67) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- Cys]-OH; (SEQ ID NO: 68), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-OH (SEQ ID NO: 69), or a pharmaceutically acceptable salt thereof, of the polypeptide according to claim 1 or 2. The polypeptide according to claim 1 or 2, which is a pharmaceutically acceptable salt.

22. The following structural formula: Ac-Arg-cyclo[Cys-D-Leu-His-D-Phe-Arg-Trp- Cys]-NH ２ ; (SEQ ID NO: 61) Ac-Arg-cyclo[Cys-D-Ile-His-D-Phe-Arg-Trp- [Cys]-NH ２ ; (SEQ ID NO: 62) Ac-Arg-cyclo[Cys-D-Tle-His-D-Phe-Arg-Trp- [Cys]-NH ２ ; (SEQ ID NO: 63), or Ac-Arg-cyclo[Cys-D-Val-His-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 10), or a pharmaceutically acceptable salt, solvate, or prodrug thereof A pharmaceutically acceptable salt.

23. In a subject in need of treating a disorder responsive to the regulation of MC4R, a method of treating a disorder responsive to the regulation of MC4R, comprising administering to the subject an effective amount of the polypeptide according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

24. The disorder responsive to the regulation of MC4R is type 1 diabetes, type 2 diabetes, obesity, insulin resistance, metabolic syndrome, male erectile dysfunction, female sexual dysfunction, non-alcoholic fatty liver disease, or non-alcoholic steatohepatitis, the method according to claim 23.

25. A pharmaceutical composition comprising the polypeptide according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in a pharmaceutically acceptable carrier. ​ ​ ​