Peptide Composition

Abstract

Polypeptide compounds that are modulators (eg, agonists and antagonists) of the melanocortin-4 receptor (MC4R), and pharmaceutical compositions comprising the same, are provided. The present invention relates to a polypeptide having the following structural formula or a pharmaceutically acceptable salt thereof: TIFF2022084844000034.tif14162 Specifically, for example, Ac-Arg-cyclo[Cys-D-Leu-His-D-Phe-Arg-Trp-Cys]-NH2; (SEQ ID NO: 61) is shown.

Description

[Technical field]

[0001] This application is a continuation of U.S. Provisional Application No. 61 / 790,469, filed March 15, 2013. No. 60 / 339,933, filed on Oct. 23, 2007, the teachings of which are incorporated herein by reference. [Background technology]

[0002] Disorders such as obesity, metabolic syndrome, insulin resistance and diabetes are becoming a national health care issue. This significantly increases costs and severely impacts the quality of life of affected individuals, their families and caregivers. The incidence of these disorders is increasing and approaching epidemic proportions. Thus, there is a need for compositions and methods for treating these disorders. . Summary of the Invention

[0003] The present invention relates to polypeptide compounds that are modulators of the melanocortin-4 receptor (MC4R). and a pharmaceutical composition containing the same.

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

[0005] or a pharma- ceutically acceptable salt thereof, During the ceremony, R 1 is -H, or C1-C6 acyl; R 2 -NR 3 R 4 -OR 5 and R 3 , R 4 and R 5 are independent of each other is H or C1-C6 alkyl; A 1 are Arg, Lys, Orn, His, Nle, Phe, Val, Leu, Trp , Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzAl or A 1 is an optionally substituted C1-C12 alkyl, an optionally substituted C6-C 18 aryl, optionally substituted C5-C18 heteroaryl, aryl moiety substituted C6-C18 aryl, optionally substituted, The aralkyl, aryl or heteroaryl moieties may be substituted. C5-C18 heteroaryl, the alkyl moiety of which is optionally substituted, a moiety selected from heteroaralkyl, 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, P ro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, A sn, Aib, or residue Y, wherein Y is Structural formula [ka]

[0006] 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 , 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 or -OH; A 4 does not exist or is Atc, Ala, QAla, Aib, Sar, Ser , Thr, Pro, Hyp, Asn, Gln, optionally substituted His, Trp, T an amino acid or residue X selected from yr, Lys, Arg, sChp, where X is The following structural formula: [ka]

[0007] and wherein the amino acid is selected from the amino acids represented by During the ceremony, R 51 and R 52 are each independently H, -CH3, phenyl, or benzyl and; R 61 , R 62 , R 63 and R 64are each independently H, -CH3, or -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 or -OH; A 5 is optionally substituted Phe, optionally substituted 1-Nal, or 2-Nal, which may be A 6 is Arg; A 7 is Trp, Any amino acid residue may be in either the L- or D-configuration, provided that: (1)A 3 and A 4 are both not non-existent; (2)A 4 is an amino acid, A 3 is neither Aib nor Gly; (3)A 4 is His, and A 5 is D-Phe or 2-Nal, A 3 teeth, Not D-amino acids or L-Ala; (4)A 2 and A 8 are selected from Cys, hCys, or Pen, If, (a)A 4 When does not exist, A 3 , but not L-His; (b)A 3 When does not exist, A 4 , but not L-His; (c)A 4 When is His, A3 is not Glu, Leu, or Lys, It is a polypeptide.

[0008] The present invention also provides a method for treating a disorder responsive to modulation of MC4R in a subject in need of treatment. The method comprises administering an effective amount of the MC4R modulation agent described herein to a subject. In certain embodiments, disorders responsive to modulation of MC4R include, but are not limited to, Type 2 diabetes, obesity, insulin resistance, metabolic syndrome, male erectile dysfunction, female sexual function Alcohol-related disorders include nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, and alcohol use disorders. These include substance abuse disorders, cachexia, inflammation and anxiety.

[0009] In certain embodiments, the compounds and compositions of the present invention bind to the melanocortin-1 receptor (M Higher levels of MC4R and melanocortin-3 receptor (MC3R) compared with melanocortin-3 receptor (C1R) The compounds and compositions of the present invention have high selectivity and efficacy. Undesirable side effects such as increased number of sperm, sexual stimulation and increased skin pigmentation Unwanted side effects can be reduced or eliminated. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] A description of an example embodiment of the present invention follows.

[0011] (term) The terminology used to define peptides is typically used in the art. The term is used in the synthesis of ribozymes, where the N-terminal amino group appears on the left and the C-terminal carboxyl group appears on the right. can be.

[0012] As used herein, the term "amino acid" refers to naturally occurring and non-naturally occurring amino acids. Unless otherwise specified, the amino acids found in the compounds described herein include both natural and non-natural amino acids. All amino acids and residues thereof may be in either the D or L configuration.

[0013] The compounds of the invention useful for practicing the methods described herein include those that contain one or more chiral It may have a center and therefore may exist in many stereoisomeric forms. All stereoisomers and mixtures thereof are within the scope of the present invention. The compounds may be separated using either preparative HPLC or columns with chiral stationary phases. or may be resolved to obtain the individual enantiomers using methods known to those skilled in the art. In addition, chiral intermediate compounds can be resolved and used to prepare chiral compounds of the invention. It may also be manufactured.

[0014] The compounds described herein may exist in one or more tautomeric forms. All tautomers and mixtures thereof are included within the scope of the present invention.

[0015] Unless otherwise defined, all technical and scientific terms used herein are and have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. All publications, patent applications, patents, and other references mentioned herein are hereby incorporated by reference. and incorporated. [Table 1] TIFF0007654587000005.tif192149 TIFF0007654587000006.tif158139

[0016] Unless otherwise specified, all abbreviations of amino acids in this disclosure (e.g., Ala) refer to amino acids. refers to a cyclic acid residue (i.e., represents the structure -NH-C(R)(R')-CO-, where R and R Each ' is independently hydrogen or an amino acid side chain (e.g., R for Ala). =CH3 and R'=-H, or R and R' may be joined to form a ring system).

[0017] The designations "Ac" or "NH2" at the polypeptide termini represent the corresponding termini, respectively. is acylated or amidated.

[0018] The phrase "covalent bond between amino acid side chains" refers to a bond between the sides of the two amino acid residues in question. This means that the chains each contain functional groups capable of forming covalent bonds with each other. Examples of such bonds include those made by Cys, hCys or Pen side chains. Disulfide bridges, which are formed by bonding the amino group of one amino acid side chain to the carboxyl group of another amino acid side chain, Amide bonds formed by carboxy groups (e.g., Asp, Glu, Lys, Orn, In an exemplary embodiment, the amino acids are each The amino acids can be selected so that a covalent bond can be formed between their side chains. When a covalent bond between the side chains is formed, the polypeptide may be cyclized. Lipeptides are identified by their structural formula or by the abbreviation "c()" or "cyclo()". For example, "-c(Cys-Cys)-" or "-cyclo(Cys-Cys)-" may be used. (Cys-Cys)-" is [ka]

[0019] On the other hand, "-c(Asp-Lys)-" or "-cyclo(Asp-Lys) )-" is the structure [ka]

[0020] Shows.

[0021] Used alone or in the form of "hydroxyalkyl", "alkoxyalkyl", "a "Alkyl" when used as part of a larger moiety, such as "alkylamine," is a specific A saturated straight or branched chain fatty acid having a number of carbon atoms, typically 1 to 12 carbon atoms. More specifically, the aliphatic group is a group having 1 to 8, 1 to 6, or 1 to 4 The term may include, for example, methyl, ethyl, n-propyl, isopropyl, ethyl, propyl, butyl ... n-propyl, n-butyl, isobutyl, tert-butyl, n-hexyl, etc. Examples are given below.

[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 radical -CN.

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

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

[0027] Used alone or as part of a larger moiety such as "aralkyl" "Aryl" as used herein refers to an alkyl group having 6 to 18 carbon atoms and one or more fused rings. The term "aryl" refers to an aromatic carbocyclic group having a cycloalkyl group or a heterocyclic group. It also includes aromatic carbocyclic ring(s) fused to a cycloalkyl group. Examples of aryl groups are: The following are examples of phenyl, benzo[d][1,3]dioxole, naphthyl, and phenanthrene. etc.

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

[0029] "Aralkyl" refers to a group in which at least one alkyl hydrogen atom is bonded to an aryl moiety, e.g., aryl. phenyl, -(CH2)2phenyl, -(CH2)3phenyl, -CH(phenyl)2, etc. It refers to an alkyl group replaced with .

[0030] Used alone or as part of a larger moiety such as "heteroaralkyl" "Heteroaryl" as used herein refers to one independently selected from nitrogen, oxygen, and sulfur. A 5- to 18-membered monocyclic, bicyclic or tricyclic heteroaromatic ring system containing up to 4 heteroatoms The term "heteroaryl" refers to a cycloalkyl group or a heterocycloalkyl group. Particular examples of heteroaryl groups include , optionally substituted pyridyl, pyrrolyl, pyrimidinyl, furyl, thienyl, imidazolyl, Zolyl, 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-oxa Diazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl, benzofuryl, [ 2,3-dihydro]benzofuryl, isobenzofuryl, benzothienyl, benzotriazo aryl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzyl Midazolyl, imidazo[1,2-a]pyridyl, benzothiazolyl, benzoxazolyl , quinolizinyl, quinazolinyl, phthalazinyl, quinoxalinyl, cinnolinyl, naphthyl Lysinyl, pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4 ,3-b]pyridyl, quinolyl, isoquinolyl, tetrazolyl, 1,2,3,4-tetrazolyl Hydroquinolyl, 1,2,3,4-tetrahydroisoquinolyl, purinyl, pteridinyl , carbazolyl, xanthenyl, benzoquinolyl, and the like.

[0031] "Heteroaralkyl" refers to an alkyl group in which at least one alkyl hydrogen atom is replaced by a heteroaryl moiety, For example, alkyl substituted with -CH2-pyridinyl, -CH2-pyrimidinyl, etc. Point.

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

[0033] "Hydroxyalkyl" and "alkoxyalkyl" refer to hydroxy and alkoxyalkyl groups, respectively. It is an alkyl group substituted with an alkoxy group.

[0034] "Amino" means -NH2, "alkylamine" and "dialkylamine" are and -NHR and -NR2, respectively, where R is an alkyl group. "Dicycloalkylamine" and "dicycloalkylamine" refer to -NHR and -NR2, respectively. where R is a cycloalkyl group. "Cycloalkylalkylamine" is -NHR R is a cycloalkylalkyl group. "Cycloalkylamine" means -N(R)2, where R is cycloalkylalkyl, and R is alkyl.

[0035] "Acyl" refers to R"-C(O)-, where R" is H, alkyl, substituted alkyl, or heteroaryl. aryl, alkyl, substituted heteroalkyl, alkenyl, substituted alkenyl, aryl, alkyla aryl, or substituted alkylaryl, which is represented in the general formula of certain embodiments as "Ac." will be done.

[0036] Suitable substituents for "alkyl", "aryl" or "heteroaryl" are as defined herein. Examples of suitable substituents are halogen, -CN, -OH. , -NH2, (C1-C4) alkyl, (C1-C4) haloalkyl, aryl, hetero Aryl, (C3-C7)cycloalkyl, (5-7 membered ring)heterocycloalkyl, -N H(C1-C6)alkyl, -N((C1-C6)alkyl)2, (C1-C6)alk oxy, (C1-C6)alkoxycarbonyl, -CONH2, -OCONH2, -NHC ONH2, -N(C1-C6)alkylCONH2, -N(C1-C6)alkylCON H(C1-C6) alkyl, -NHCONH(C1-C6) alkyl, -NHCON(( C1-C6) alkyl)2, -N(C1-C6) alkylCON((C1-C6) alkyl -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) Alky -CO(C1-C6)alkyl, -OCO(C1-C6)alkyl, -C(O) O(C1-C6)alkyl, -OC(O)O(C1-C6)alkyl, -C(O)H or 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. Within the framework of the invention, the term "substitution" includes the situation where a hydrogen atom is replaced with a deuterium atom. where p is an integer having a value of 1 or 2.

[0037] Suitable substituents for substituted Phe include 1-5 substituents on any aromatic carbon. groups, and the substituents are F, Cl, Br, I, -CH3, -OH, -CN, amine, - 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 1-3 aryl groups on any substitutable ring atom. The substituents include F, Cl, Br, I, -CH3, -OH, -CN, and amine. Examples are 1-methyl- Examples include histidine and 3-methyl-histidine.

[0039] "(amino acid) n " means that the amino acid is repeated n times. For example, the designations "(Pro)2" or "(Arg)3" refer to a proline residue or an arginine residue. This means that the arginine residue is repeated two or three times, respectively.

[0040] Pharmaceutically acceptable salts of the polypeptide compounds disclosed herein are included in the present invention. For example, an acid salt of a compound containing an amine or other basic group can be prepared by reacting this compound with an appropriate functional group. The aniline may be obtained by reacting with an organic or inorganic acid, and is pharma- ceutical acceptable. Ionic salt forms are obtained. Examples of anionic salts are acetate, benzenesulfonate, Benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsilate , carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate , Esylate, Fumarate, Gluceptate, Gluconate, Glutamate, Glycolyate Glycollylarsanilate, Hexylresorcylic Acid Salt, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, milk Salts, lactobionates, malates, maleates, mandelates, mesylates, methylates sulfate, mucate, napsylate, nitrate, pamoate, pantothenate Tothenates, phosphates / diphosphates, polygalacturonates, salicylates, stearates Salt, acetate, succinate, sulfate, tannate, tartrate, tetraoctyl, tocopherol These include trifluoroacetate, triethiodide and trifluoroacetate.

[0041] Salts of compounds containing an acid functional group can be prepared by reacting with a suitable base. Such pharma- ceutically acceptable salts can be prepared using bases and pharma- ceutically acceptable compounds. The thiones may be provided, as well as alkali metal salts (especially sodium and potassium), alkali Earth metal salts (especially calcium and magnesium), aluminum salts and ammonium salts salts, and physiologically acceptable organic bases (e.g., trimethylamine, triethylamine, amine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N,N '-Dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxy (Diethyl)amine, Tri-(2-hydroxyethyl)amine, Procaine, Dibenzylpyridine Peridine, dehydroabietylamine, N,N'-bisdehydroabietylamine, glutamate amine, N-methylglucamine, collidine, quinine, quinoline) and basic amino acids (e.g., lysine and arginine).

[0042] The disclosed compounds can be administered as part of a pharmaceutical composition in combination with an acceptable pharmaceutical carrier. The formulation of the compound to be administered may vary depending on the route of administration selected. Suitable pharmaceutical carriers may be in the form of a liquid or liquid formulation (e.g., solution, emulsion, capsule). May contain non-interacting inactive ingredients. Standard pharmaceutical compounding techniques may be used. For example, see Remington's Pharmaceutical Sciences s, Mack Publishing Company, Easton, PA 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, etc. Methods for encapsulating compositions (e.g., hard gelatin or cyclodextran encapsulation) Such methods, such as those described in "Cont Rolled Release of Biological Active Agent John Wiley and Sons, 1986).

[0043] As used herein, the term "disorders responsive to modulation of the melanocortin-4 receptor" refers to The phrase refers to any drug that can be treated by activating (stimulating) or inhibiting MC4R. Examples of such disorders are described in detail below.

[0044] As used herein, the term "modulator" refers to a substance that affects a biological function and inhibits the activity of a target receptor. Examples of modulators include full agonists, partial agonists, and , neutral antagonists and inverse agonists.

[0045] As used herein, the term "agonist" refers to an agonist that acts on its target, here MC4R, and Upon interaction (e.g., binding), it suppresses the signaling activity of MC4R from its basal level. Agonist refers to any naturally occurring or synthetic chemical compound listed below. is a superagonist (i.e., an agonist that acts greater than the endogenous agonist for the target receptor). Compounds capable of producing a maximal response and therefore having greater than 100% efficacy), Full agonists (i.e., compounds that elicit a maximal response following receptor occupancy and activation) ) or partial agonists (i.e., capable of activating the receptor but not the full extent of the receptor system) Examples of MC4R agonists include compounds that are unable to induce a large response to MC4R. The details are described below.

[0046] As used herein, the term "antagonist" refers to an antagonist that inhibits the activity of its target, here MC4R. Upon interaction with (e.g., binding to) the agonist compound in a dose-dependent manner, Refers to any chemical compound that blocks signaling activity with MC4R. As used herein, the term "inverse agonist" refers to an agonist that inhibits the activity of its target, here M Upon interaction with (e.g., binding to) C4R, it upregulates the activation of MC4R in a dose-dependent manner. This refers to any chemical compound that reduces the basal level of signal transmission activity.

[0047] As used herein, an "effective amount" refers to an amount effective to treat or prevent a targeted disorder. An effective amount refers to an amount of a therapeutic agent or combination of therapeutic agents sufficient to treat an individual. Examples of effective amounts include: Typically, the range is about 0.0001 mg / kg body weight to about 500 mg / kg body weight. An exemplary range is from about 0.0001 mg / kg to about 500 mg / kg of body weight. Effective amounts may range from about 0.005 mg / kg to about 500 mg / kg. In examples, the range may be from about 0.0001 mg / kg to about 5 mg / kg. In other examples, the effective amount is about 0.01 mg / kg body weight to 50 mg / kg body weight, or 0 It may be in the range of 0.01 mg / kg to 20 mg / kg of body weight.

[0048] As used herein, the term "subject" refers to a mammal, preferably a human, e.g. For example, pet animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, Veterinary procedures for animals (e.g. horses) and laboratory animals (e.g. rats, mice, guinea pigs) It can also mean an animal in need of treatment.

[0049] As used herein, the term "second agent" refers to a peptide or peptides described herein. In combination, they may enhance the therapeutic effects of the peptides described herein alone, or exhibit synergistic effects (i.e., greater than additive effects) with the peptides described herein. As used herein, the term "combined active pharmaceutical ingredient" includes any active pharmaceutical ingredient (API) that exhibits a combined effect. When used in combination, "enhanced therapeutic effect" refers to an improved therapeutic profile through means other than synergy. Examples of enhanced therapeutic effects include effective dosages of the peptides described herein. and extending the therapeutic window of the peptides described herein. One or more second drugs may be administered. Examples of second drugs are detailed below. Please write down.

[0050] The second agent may be administered prior to, simultaneously with, or after administration of the peptides described herein. Thus, the peptides described herein and the second agent may be administered in a single combination. They may be administered together as a combination or in separate formulations, e.g., simultaneously or sequentially. For example, the peptides described herein and the second agent may be administered sequentially in separate compositions. and the peptides described herein may be administered before or after the second therapeutic agent. In addition, the peptides described herein and the second agent can be administered in a similar dosing schedule. For example, the peptides described herein may or may not be administered in a dose-dependent manner. and a second therapeutic agent, the peptide described herein being more frequently administered than the second therapeutic agent. or vice versa, where the second therapeutic agent is a peptide as described herein. They may have different half-lives and / or different Finally, the peptides described herein may be followed by a second The sequential administration of both therapeutic agents may result in further enhanced therapeutic efficacy. Either the peptides or the second agent described herein can be administered acutely or chronically. may be administered to

[0051] In a method or composition of the invention, a compound having a first amount of MC4R modulator activity is or a pharma- ceutically acceptable salt thereof together with a second amount of at least one second agent. In one embodiment, the effective amount can be achieved by administering The peptides described and the second agent are each administered in an effective amount (i.e., when administered alone). In another embodiment, the compounds described herein are administered in an amount that is therapeutically effective for each individual compound. The peptides described and the second agent alone do not provide a therapeutic effect (subtherapeutic doses). In yet another embodiment, the peptides described herein are administered in an amount of 0.1 mg / kg / day or 100 mg / kg / day. The first agent may be administered in an effective amount, while the second agent is administered in a sub-therapeutic amount. In yet another embodiment, the peptides described herein are administered in sub-therapeutic amounts. In an exemplary embodiment, the second agent is administered in an effective amount. The combination of the described peptides with a second agent is similar to that of the peptides described herein alone. or a second agent that exhibits enhanced therapeutic efficacy or a synergistic effect. Shows.

[0052] The presence of synergy should be determined using methods appropriate for evaluating drug interactions. Suitable methods include, for example, the Sigmoid-Emax formula (Holford, N.H.G. and Scheiner, L.B., Clin Pharmacokin. 6:429-453(1981)), Loewe additivity formula (Loewe, S. and and Muischnek, H., Arch. Exp. Pathol. Pharmacol. 114:313-326(1926)) and the formula for the median effect (Chou, T. C. and and Talalay, P., Adv. Enzyme Regul. 22:27-55(1 Each of the above-referenced equations is applied to experimental data to obtain the Corresponding graphs can be generated to assist in assessing the combination effect. The corresponding graphs associated with the above referenced formulas are concentration-effect curves, isoborogles, Lamb curve and combination exponential curve.

[0053] As used herein, "treating" refers to reducing or preventing the progression of clinical symptoms associated with the targeted disorder. to partially or substantially slow, inhibit, or prevent For example, "treating" can mean treating a condition (e.g., as measured by body mass index, BMI). ) Partial or complete weight loss; Clinical symptoms or indicators associated with obesity, e.g. II Diabetes, prediabetes, blood levels of hemoglobin A1C (HblAc) higher than 6% Improves or improves hyperinsulinemia, hyperlipidemia, insulin resistance, glucose intolerance, etc. slowing, inhibiting, or preventing the progression of obesity and obesity-related symptoms; and / or partially or completely delay the onset or progression of obesity or obesity-related symptoms. partially or substantially achieve one or more of the results of inhibiting, preventing, or preventing Slowing, inhibiting, or preventing the progression of obesity includes, for example, and slowing, inhibiting, or preventing the progression of obesity in a subject of normal weight. The term "treat" further includes treating coronary artery disease, stroke and diabetes associated with metabolic syndrome. Partially or completely reducing the risk of diseases (e.g., type 2) and metabolic syndrome-related Clinical symptoms or signs of metabolic syndrome, such as any of the five indicators listed above, that For example, the term "treat" includes reducing or ameliorating insulin resistance, sucrose Parameters related to metabolic syndrome, including clearance, and cardiovascular, including heart rate and blood pressure parameters of vascular disease, joint disease, inflammation, sleep apnea, binge eating, and other feeding disorders including bulimia Slowing, inhibiting, or preventing the progression of eating disorders; as an adjunct to weight loss surgery "Preventive treatment" includes targeted therapy, including weight loss therapy prior to orthopedic surgery, and adjunctive weight loss therapy prior to orthopedic surgery. Prevent, inhibit, or reduce the onset of a disorder before it develops clinically. This refers to the measures taken to

[0054] (Disorders responsive to MC4R regulation) Examples of disorders responsive to modulation of MC4R include acute and chronic inflammatory diseases, e.g. Systemic inflammation, inflammatory bowel disease, brain inflammation, sepsis and septic shock; autoimmune components diseases accompanied by weight gain, such as rheumatoid arthritis, gouty arthritis and multiple sclerosis; Metabolic diseases and medical conditions, such as obesity, eating disorders and Prader-Williams syndrome syndrome; metabolic diseases and medical conditions accompanied by weight loss, e.g., anorexia, bulimia, AI DS Wasting, cachexia, cancer cachexia and wasting in frail elderly; diabetes and diabetes-related associated conditions and complications of diabetes, e.g., retinopathy; neoplastic growths, e.g., skin cancer and and prostate cancer; reproductive or sexual medical conditions, such as endometriosis and uterine prostate cancer in women; Blood, sexual dysfunction, erectile dysfunction and decreased sexual response in women; Diseases or conditions resulting from, for example, organ transplant rejection, ischemia and reperfusion injury, spinal cord injury, Treatment of wounds and to promote wound healing, as well as chemotherapy, radiation therapy, palliative care, weight loss caused by permanent immobilization or dialysis; cardiovascular disease or condition , e.g., hemorrhagic shock, cardiogenic shock, hypovolemic shock, cardiovascular disorders and cardiac cachexia;pulmonary diseases or conditions, e.g., acute respiratory distress syndrome, chronic obstructive pulmonary disease, asthma and pulmonary fibrosis; disorders that increase immune tolerance and fight off attacks on the immune system, e.g. those associated with certain allergies or organ transplant rejection; dermatological diseases and conditions, e.g. , psoriasis, reduction of skin pigmentation, treatment of acne, keloid formation and skin cancer; behavioral, central nervous Neurological or neurological conditions and disorders, such as anxiety, depression, memory disorders and memory function disorders. Alcohol consumption, alcoholism and / or or alcoholism-related conditions and diseases; and renal conditions or diseases, e.g., Examples include the treatment of renal cachexia or natriuresis. release of aldosterone, synthesis and release of aldosterone, body temperature, blood pressure, heart rate, vascular tone, and cerebral blood flow , blood sugar level, bone metabolism, bone formation or growth, ovarian weight, placental growth, prolactin and secretion of FSH, fetal growth in the uterus, parturition, spermatogenesis, secretion of sebum and pheromones, Normalize the target, including neuroprotection and neurodevelopment, and regulating motivation, learning, and other behaviors Further examples include binge eating, bulimia, and homeostatic activity. or other eating disorders.

[0055] In an exemplary embodiment, the disorder responsive to modulation of the MC4R receptor is type 1 diabetes, type 2 diabetes mellitus, diabetes, obesity, insulin resistance, metabolic syndrome, cardiovascular disease, or low-density lipoprotein / High-density lipoprotein / triglyceride imbalance, nonalcoholic fatty liver disease and substance abuse This is an obstacle to use.

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

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

[0058] (Diabetes and Related Disorders) In an exemplary embodiment, the subject treated by the methods provided herein is You are at increased risk of having or developing a diabetes-related disorder. The disorders that affect the immune system include diabetes mellitus (type 1 (OMIM 222100) and type 2 (OMIM 125 853)), referring to insulin resistance and metabolic syndrome.

[0059] In an exemplary embodiment, the subject being treated is a patient suffering from diabetes (type 1 or type 2), insulin resistance, In an exemplary embodiment, the disorder is diabetes, e.g., type 2 In an exemplary embodiment, the subject is a World Health Organisation ization and the International Diabetes Federation "Definition and Diagno" published by ion in 2006 sis of diabetes mellitus and intermediat e hyperglycaemia" (incorporated herein by reference in its entirety). ) or are at increased risk of developing type 2 diabetes mellitus. In certain embodiments, the diabetic subject has a fasting plasma glucose of 126 mg / dL or greater. or 2-hour plasma glucose (2 hours after oral administration of 75 glucose) was 2 In an exemplary embodiment, the diabetic or prediabetic subject has a high The level of glycated hemoglobin is shown, e.g., 4.5, 4.6, 4.7, of total hemoglobin. 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.8, 6.0, 6.2, 6.4, 6.6, 6.8, 7.0, 7.2, 7.4, 7.6% or more In an exemplary embodiment, the glycated hemoglobin is expressed in one or more genes in Table 1 below. is a genetic polymorphism (e.g., a polymorphism that results in altered expression levels, e.g., near one or more genes) For example, by increasing or decreasing expression levels and / or by causing variations in the coding sequence. A diabetic or prediabetic subject may be identified or further characterized by the polymorphisms present in the You may do so. [Table 2]

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

[0061] In an exemplary embodiment, the subject treated by the methods of the invention has type I diabetes. In an exemplary embodiment, a subject with type I diabetes is diagnosed with: For example, fasting C-peptide levels of less than about 1.0 nmol / L, e.g., less than about 1.2 nmol / L < 1.1 nmol / L, < 1.0 nmol / L, 0.9 nmol / L Less than 0.8nmol / L, Less than 0.7nmol / L, Less than 0.6nmol / L, 0 Less than 0.5 nmol / L, less than 0.4 nmol / L, or even less, e.g., 0 < .33 nmol / L, < 0.25 nmol / L, < 0.2 nmol / L, or In an exemplary embodiment, the C-peptide Levels were measured after an oral glucose challenge (2 h after 75 g glucose administration). after 1 hour), less than 0.54 nmol / L, e.g., less than 0.50 nmol / L, 0.45 nmol / L less than 0.40nmol / L, less than 0.35nmol / L, less than 0.30nmol Less than l / L, Less than 0.25nmol / L, Less than 0.20nmol / L, 0.15nmol / L or an increase of less than 0.10 nmol / L is detected. 10-125 mg / dL) or impaired glucose tolerance (2 hours after a 75 g glucose challenge) (after: 140-199 mg / dL) to assess decreased β-cell function in subjects with type 1 diabetes In an exemplary embodiment, type 1 diabetes is , autoantibodies against islet cell antigens and / or insulin, e.g., 65 kDa GAD (OMIM 138275) and / or the phosphatase-associated IA-2 molecule The disease is identified or further characterized by the presence of associated autoantibodies.

[0062] (insulin resistance) In an exemplary embodiment, the disorder is "insulin resistance," any method known in the art. The disorder may be identified by any means and is characterized by a reduced ability of insulin to lower blood glucose levels. In an exemplary embodiment, insulin resistance is associated with a transcription factor receptor 1 (TRT1), a transcription factor receptor 2 (TCR2), a transcription factor receptor 3 (TCR3), a transcription factor receptor 4 (TCR4), a transcription factor receptor 5 (TCR5), a transcription factor receptor 6 (TCR6), a transcription factor receptor 7 (TCR7), a transcription factor receptor 8 (TCR8), a transcription factor receptor 9 (TCR9), a transcription factor receptor 10 (TCR10), a transcription factor receptor IM 142410; see, for example, polymorphism 0011), PPP1R3A (OMIM 60 0917; see, for example, polymorphisms 0001, 0003), PTPN1 (OMIM 1768 85; see e.g., polymorphism 0001), ENPP1 (OMIM 173335; see e.g., see polymorphism 0006), IRS1 (OMIM 147545; see e.g., polymorphism 0002 (see references), EPHX2 (OMIM 132811; see for example polymorphism 0001), leptin (OMIM 164160, see e.g., polymorphisms 0001 and 0002), leptin receptor Polymorphisms (OMIM 601007, e.g., polymorphisms 0001, 0002, 0004 and 00 05), or insulin receptor (INSR, OMIM 147670, e.g. by the presence of one or more polymorphisms in one or more genes (see polymorphisms 0001-0037) For example, polymorphisms that result in altered expression levels, e.g., polymorphisms that increase or decrease expression levels. and / or multiple mutations resulting in variations in the coding sequence of the gene product, e.g., protein. The antigen-binding protein is identified or further characterized by a specific antigen-binding protein (ApG) or a specific antigen-binding protein (AbG).

[0063] (metabolic syndrome) In an exemplary embodiment, the disorder is metabolic syndrome. "Metabolic syndrome" is a condition in which coronary artery disease, stroke, and type 2 diabetes occur together and these risks American Heart Association and the National Heart, Lung and Blood In According to the Institute, Metabolic Syndrome, also known as Syndrome X, is defined as a condition in which three or more of the following conditions occur: Present when symptomatic.

[0064] (1) Blood pressure 130 / 85 mmHg or higher, (2) Fasting blood glucose level of 100 mg / dL or higher; (3) Large waist circumference (length around the waist): -Males - 40 inches and up -Women-Over 35 inches (4)Low HDL cholesterol: - Men - Lower than 40 mg / dL -Women - Lower than 50 mg / dL (5) Triglycerides are 150 mg / dL or higher.

[0065] Metabolic syndrome was evaluated by measuring blood pressure, blood glucose, HDL cholesterol, and LDL cholesterol. It can be diagnosed by testing blood glucose levels, total cholesterol and triglyceride levels. can.

[0066] In an exemplary embodiment, the subject has central obesity (for women, waist circumference ≧8 0 cm; ≥ 90 cm for Asian men, including South and Central American men, etc. ≥ 94 cm for all men, and BMI > 30 kg / m 2 , elevated triglycerides (≧ 150 mg / dL, or specific treatment for this lipid abnormality), lowering HDL cholesterol ( <40 mg / dL in men, <50 mg / dL in women, or specific treatment for this lipid abnormality placement), elevated blood pressure (sBP ≥ 130mmHg or dBP ≥ 85mmHg, or already Treatment of diagnosed hypertension), or elevated fasting plasma glucose (FPG ≥ 100 mg / mL) dL, or previous diagnosis of type 2 diabetes), and combinations thereof. In the present invention, the subject to be treated by the method provided herein is an Internatio The "T" published in 2006 by the National Diabetes Federation he IDF consensus worldwide definition of the metabolic syndrome," which is incorporated herein by reference in its entirety. Having or at risk for metabolic syndrome, as defined by the International Clinical Trials and Clinical Trials (ICT-CTA) That is, the subject has central obesity (as described above and / or BMI>30 kg / m 2 ), and increased triglycerides and decreased HDL cholesterol , elevated blood pressure, or elevated fasting plasma glucose. In certain embodiments, the metabolic syndrome is diagnosed in a subject by determining a mutation at 3q27 (e.g., OMIM 605 552) and / or 17pl2 (see, e.g., OMIM 605572) or further characterized by the presence of a mutation at a locus selected from The characteristics are determined.

[0067] (Disorders caused by MC4R mutations) The present invention relates to a method for the treatment of α-melanocortin stimulating hormone (α-MSH) The present invention relates to a method for treating a disorder in a subject resulting from a weakened C4R response. The method includes administering an effective amount of a melanocortin-4 receptor (MC4R) agonist. In an exemplary embodiment, the subject is administered a hormone that stimulates α-melanocortin (α Heterozygous carriers of an MC4R mutation that weakens the response of MC4R to β-MSH Heterozygotes retain the ability to respond to natural ligands of MC4R, and thus Treatment of MC4R-related disorders in telozygotes with MC4R agonists Treatment is independent of knowledge of the type of MC4R mutation.

[0068] In an exemplary embodiment, the disorder is obesity, e.g., MC4R-associated obesity. In an exemplary embodiment, the disorder is metabolic syndrome.

[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 severe 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 portions of which are incorporated herein by reference. are incorporated by reference into the book.

[0071] Additional mutations potentially causing obesity in humans include those reported by 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, 2010-06-08;49(2 2):4583-600, the relevant portions of which are incorporated herein by reference. As shown, 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.

[0072] Further examples of mutations potentially causing obesity in humans are listed in the Online Me Human genes and The database of genetic disorders and genetic disorders, http: / / omim.org / entry / 1 The URL for accession no. 155541 (MC4R) (more precisely, accession no. 1 55541.0001~155541.0023). Representative examples include 4- BP DEL, NT631;4-BP INS, NT732;TYR35TER;ASP 37VAL;SER58CYS;ILE102SER;ASN274SER;1-BP INS, 112A;4-BP DEL, 211CTCT;ILE125LYS;ALA1 75THR;ILE316SER;TYR287TER;ASN97ASP;15-BP DEL (delta 88-92 codons); and SER127LEU. The relevant portions of the M database are incorporated herein by reference.

[0073] In an exemplary embodiment, a mutation in MC4R reduces the signaling activity of MC4R. Hold.

[0074] Mutations in the genomic sequence encoding MC4R are detected by methods well known to those skilled in the art. For example, a genomic sequence can be synthesized using a nucleotide primer, e.g., 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 Investigation, 2 The primers described in vol. 106(2), pp. 253-262, July 2000, were used. The cloned sequences were then sequenced using a commercially available sequencer and software. can be used for analysis.

[0075] The activity of MC4R can be measured by methods well known to those skilled in the art. Cells were transiently transfected with cloned MC4R DNA and transfected. The transfected cells are contacted with an MC4R agonist (e.g., α-MSH) to induce MC4R The intracellular level of cAMP, the second messenger of Journal of Endocrinology (2010)207, pp.17 MC4R signal transduction is measured by electrochemiluminescence assay as determined in Example 7-183. The decrease in activity is due to a decrease in the amount of cAMP produced by wild-type C4R in response to a given agonist. The intracellular levels of MC4R were determined by comparing the intracellular levels produced by mutant MC4R. It can be acknowledged.

[0076] MC4R modulators (e.g., agonists) can be used to inhibit, for example, the natural agonist of MC4R. The present invention may also be used to treat patients suffering from other disorders, including reduced availability of Patient examples include key regulators of leptin-dependent pathways or proopiomelanocortin processing. Individuals who are heterozygous or homozygous for a mutation in a gene (Nature Clinical Practice Endocrinology and Met abolism, 2006;2;6;318 and NEng J Med:2007;3 56;3;237, Nature Genetics, 1998, 155;Cell M etabolism, 2006;3;135;Annals Acad Med, 200 9, 38;1;34), or mutations in genes encoding prohormone convertases Examples include individuals with

[0077] (Mode of administration) Compounds of the invention useful for carrying out the methods described herein, or their pharma- ceutical uses, Administration of an acceptable salt or compound or pharmaceutical salt may be continuous, hourly, daily, or in combination with other suitable methods. 4 times, 3 times a day, 2 times a day, 1 time a day, 1 time every other day, 2 times a week, 1 time a week, 2 Once a week, once a month, or once every two months, or for a longer period of time, or some other intermediate dosing regimen.

[0078] As an example of administration of a compound of the invention, or a composition comprising a compound or pharmaceutical salt of the invention, Examples of peripheral administration include oral, subcutaneous, intraperitoneal, intramuscular, and intravenous. These include intravenous, rectal, transdermal, buccal, sublingual, inhalation, pulmonary, or nasal modes of administration.

[0079] (Combination Therapy) One or more other medicamentously active agents may be used to treat any disorder responsive to modulation of MC4R. The present invention provides a method for treating a patient suffering from atopic dermatitis, comprising administering the method in combination with a compound having a specific activity (a "second agent") as described herein. Such combined administration may be carried out in accordance with the methods described herein. The present invention may also be carried out by administering a single dose of one or more peptides that are capable of inhibiting the inflammatory process and one or more second drugs. Commonly, for example, such unit dosage forms include tablets, capsules, sprays, inhalation powders, etc. , injectable liquid, etc. Alternatively, the combination administration may be in two different dosage forms. In one embodiment, a dosage form may be administered containing one or more peptides as 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] In addition to, or instead of, one or more of the peptides described herein, Obesity and / or overweight-related diseases, disorders and / or conditions (e.g., insulin glucose tolerance; impaired glucose tolerance; type 2 diabetes; metabolic syndrome; dyslipidemia (including hyperlipidemia); hypertension Cardiac disorders (e.g., coronary heart disease, myocardial infarction); cardiovascular disorders; nonalcoholic fatty liver disease (including non-alcoholic steatohepatitis);joint disorders (including secondary arthritis);gastroesophageal reflux sleep apnea; atherosclerosis; stroke; large and small vessel disease; adiposity (e.g. and one or more second agents useful in the treatment of chronic obstructive pulmonary disease (COPD), such as pulmonary disease (e.g., liver disorders); gallstones; and gallbladder disorders. may be combined with

[0084] (Second drug) The one or more second agents may, for example, be selected from the following:

[0085] Insulin and insulin analogues; Sulfonylureas (e.g., glipizide) and prandial glucose regulators ("short-acting meglitinides (sometimes called "type-II secretagogues"), e.g., meglitinides (e.g., repaglinides) insulin secretagogues, including nateglinide and nateglinide; Drugs that enhance the action of incretins: incretins, incretin mimetics, incretins Drugs that improve ginseng 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., thiazolidinone Lysindiones (e.g., pioglitazone and rosiglitazone) and PPARα,γ and insulin sensitizers, including agents having any combination of δ activity; Agents that regulate hepatic glucose balance, such as biguanides (e.g. metformin) , Fructose 1,6-bisphosphatase inhibitor, Glycogen phosphorylase inhibitor agents, glycogen synthase kinase inhibitors and glucokinase activators; Medications designed to reduce / delay the absorption of glucose from the intestine, e.g., α-glucose Glucosidase inhibitors (e.g., miglitol and acarbose); Drugs that antagonize the action of glucagon or reduce glucagon secretion, such as amylin Analogues (e.g., pramlintide); Agents that prevent glucose reabsorption by the kidneys, e.g., sodium-dependent glucose transporters sodium glucosamine 2 (SGLT2) inhibitors (e.g., dapagliflozin); Drugs designed to treat the complications of long-term hyperglycemia, such as aldose reductase inhibitors (e.g., epalrestat and ranirestat); Medications used to treat complications associated with microangiopathy; Antihyperlipidemic agents, such as HMG-CoA reductase inhibitors (statins, e.g., rosacea Vastatin) and other cholesterol-lowering drugs; PPARα agonists (fibrates, e.g., gemfibrozil and fenofib raat); Bile acid sequestrants (e.g., cholestyramine); Cholesterol absorption inhibitors (e.g., plant sterols (i.e., phytosterols) , synthetic inhibitors); Cholesteryl ester transfer protein (CETP) inhibitors; an inhibitor of the ileal bile acid transport system inhibitors (IBAT inhibitors); Resins to which bile acids bind; Nicotinic acid (niacin) and its analogues; Antioxidants, such as probucol; omega-3 fatty acids; Adrenergic receptor antagonists, such as beta blockers (e.g., atenolol) ol), alpha blockers (e.g., doxazosin), and mixed alpha / beta blockers (e.g., Antihypertensives, including, for example, labetalol; Adrenergic receptor agonists, including alpha-2 agonists (e.g., clonidine) to; Angiotensin-converting enzyme (ACE) inhibitors (e.g., lisinopril), calcium channel blockers Channel blockers, e.g., dihydropyridines (e.g., nifedipine), phenylalanine alkylamines (e.g., verapamil) and benzothiazepines (e.g., diltiazem) ; Angiotensin II receptor antagonists (e.g., candesartan); aldosterone ron receptor antagonists (e.g., eplerenone); Centrally acting adrenergic agents, e.g., central alpha agonists (e.g., clonidine); and diuretics (e.g., furosemide); Antithrombotic agents, e.g., fibrinolysis activators; hemostatic modifiers, including thrombin antagonists Decorative substances; Factor VIIa inhibitors; anticoagulants, e.g., vitamin K antagonists (e.g., Warfarin Heparin and its low molecular weight analogues, factor Xa inhibitors and direct thrombin inhibitors. antiplatelet agents, e.g., cyclooxygenase inhibitors drugs (e.g., aspirin), adenosine diphosphate (ADP) receptor inhibitors (e.g., clotting pidogrel), phosphodiesterase inhibitors (e.g., cilostazol), glycoproteins IIB / IIA inhibitors (e.g., tirofiban) and adenosine reuptake inhibitors (e.g., e.g., dipyridamole); Anti-obesity agents, such as noradrenergic agents (e.g., phentermine) and serotonin receptor antagonists. Tonic agents (e.g., sibutramine), pancreatic lipase inhibitors (e.g., orlistat ), microsomal transport protein (MTP) modifier, diacylglycerol acyltransferase DGAT inhibitors and cannabinoid (CB1) receptor antagonists appetite suppressants (e.g., ephedrine), including acetaminophen (e.g., rimonabant); Feeding behavior regulators, such as orexin receptor modulators and melanin-concentrating hormone (M CH)modifier; Neuropeptide Y (NPY) / NPY receptor modulators; Pyruvate dehydrogenase kinase (PDK) modulators; Serotonin receptor modulators; leptin / leptin receptor modulators; ghrelin / ghrelin receptor modulators; drugs that enhance beta cell function; Drugs that stimulate energy expenditure (e.g., β-adrenergic stimulants, UCP-1 agonists) sto, brown fat modulators and stimulators); Agents that induce the lysis of fat cells (e.g., antibodies); Nicotine or nicotine withdrawal aids; Estrogens, natural or synthetic modulators of the estrogen receptor; μ-opioid receptor modulators; and Monoamine transporter regulators, such as selective serotonin reuptake inhibitors (SSRIs) ( For example, fluoxetine, noradrenaline reuptake inhibitors (NARIs), noradrenaline Serotonin-reuptake inhibitors (SNRIs), triple monoamine reuptake blockers Carbapenem (e.g., tesofensine) and monoamine oxidase inhibitors (MAOIs) (e.g., For example, toloxatone and amiflamine), or a pharma- ceutically acceptable salt thereof.

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

[0087] Isolated Polypeptides of the Invention In an exemplary embodiment, the isolated polypeptide (eg, an agonist of MC4R) is , Formula (I) or a pharma- ceutically acceptable salt thereof:

[0088] An isolated polypeptide having the following structural formula (I): [ka]

[0089] or a pharma- ceutical acceptable salt thereof; During the ceremony, R 1 is H, or C1-C6 acyl; R 2 -NR 3 R 4 -OR 5 and R 3 , R 4 and R 5 are independent of each other is H or C1-C6 alkyl; A 1 are Arg, Lys, Orn, His, Nle, Phe, Val, Leu, Trp , Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzAl or A 1 is an optionally substituted C1-C12 alkyl, an optionally substituted C6-C 18 aryl, optionally substituted C5-C18 heteroaryl, aryl moiety substituted C6-C18 aryl, optionally substituted, The aralkyl, aryl or heteroaryl moieties 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 or -OH; A 4 does not exist or is Atc, Ala, QAla, Aib, Sar, Ser , Thr, Pro, Hyp, Asn, Gln, optionally substituted His, Trp, T an amino acid residue selected from yr, Lys, Arg, sChp, or residue X, where X has the following structural formula: [ka] TIFF0007654587000013.tif55141

[0091] and wherein the amino acid is selected from the amino acids represented by During the ceremony, R 51 and R 52 are each independently H, -CH3, phenyl, or benzyl and; R 61 , R 62 , R 63 and R 64 are each independently H, -CH3, or -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 or -OH; A 5 is optionally substituted Phe, optionally substituted 1-Nal, or 2-Nal, which may be A 6 is Arg; A 7 is Trp, Any amino acid residue may be in either the L- or D-configuration.

[0092] In an exemplary embodiment, A 3 is absent or Ala, Tle, Val, Le u, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gl an amino acid selected from among n, Sar, Gly, Asn, or Aib; 4 exists Not present or 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 has the structure formula [ka]

[0093] The amino acid is selected from the amino acids represented by

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

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

[0096] The amino acid residue is 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). It is.

[0098] In an exemplary embodiment, A 3 and A 4 are both non-existent. The remaining values ​​of the variables and preferred values ​​are as defined above and below for formula (I).

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

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

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

[0102] In an exemplary embodiment, (1) A 3 and A 4 are not non-existent;(2)A 4 is an amino acid, A 3 is neither Aib nor Gly; (3) A 4 His A 5 is D-Phe or 2-Nal, A 3 is a D-amino acid, but is it an L-amino acid? (4) A 2 and A 8 are selected from Cys, hCys, or Pen, If selected, (a)A 4 When does not exist, A 3 (b) A, but not L-His; 3 When does not exist, A 4 (c) A, but not L-His; 4 When is His, A 3 is not Glu, Leu, or Lys. The remaining values ​​and preferred values ​​of the variables are given by the formula ( I) as defined above and below.

[0103] In another embodiment, the polypeptide of formula (I), A 4 is an L-amino acid. In the embodiment of the present invention, 4 The remaining values ​​and preferred values ​​of the variables are as set forth in formula (I). As defined above and below.

[0104] In an exemplary embodiment, A 5 is optionally substituted 1-Nal or It may be optionally substituted 2-Nal, for example, optionally substituted D-2-Nal. 5 is any of the five aromatic carbons: F, CI, Br, I, -CH3, -OH, -CN , an amine, -NO2, or -OCH3.

[0105] In a further embodiment, the polypeptide of formula (I), A 5 is an optionally substituted D- Phe. 5 is any of the five aromatic carbons: F, CI, Br, I, -CH3 , -OH, -CN, amine, -NO2, or -OCH3. A 5 Suitable examples of the amino acid sequence 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 The variable is a D-amino acid selected from Phe(3',4',5'-triF). The remaining values ​​and preferred values ​​are as defined above and below for formula (I). .

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

[0107] In yet another embodiment, the polypeptide of formula (I), A 4 At any suitable position, F , CI, Br, I, -CH3, -OH, -CN, amine, -NO2, benzyl or -O and CH3. The remaining values ​​of the variables and and preferred values ​​are as defined above and below for formula (I).

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

[0109] In another embodiment, the polypeptide of the invention has the following structural formula: [ka] TIFF0007654587000017.tif34141

[0110] or a pharma- ceutically acceptable salt thereof.

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

[0112] or a pharma- ceutically acceptable salt thereof.

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

[0114] or a pharma- ceutically acceptable salt thereof.

[0115] In yet another embodiment, the polypeptide of the present invention is a polypeptide represented by formula (I): Including Petit, A 4 are Atc, Ala, QAla, Aib, Sar, Ser, Thr, Pro, Hyp, Asn, Gln, substituted His, Trp, Tyr, Lys, Arg , sChp, or residue X. Examples of such peptides The compound has the following structural formula: 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(pF)-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-XD-Phe-Arg-Trp-Cy s]-NH2, a peptide represented by any one of (SEQ ID NO: 44), or This includes pharma- ceutically acceptable salts.

[0116] In an exemplary embodiment, the polypeptide of the 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 This includes pharma- ceutically acceptable salts.

[0117] In another embodiment, the polypeptide of the present invention is a polypeptide represented by formula (I): Including A 3 are Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, From Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, or Aib A is a selected amino acid; 4 Atc, Ala, QAla, Aib, Sar, Se r, Thr, Pro, Hyp, Asn, Gln, substituted His, Trp, Tyr, L ys, Arg, sChp, or residue X. An example of a polypeptide has the following structural formula: Ac-Arg-cyclo[Cys-Val-Gln-D-Phe-Arg-Trp-Cy s]-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-Ar g-Trp-Cys]-NH2 (SEQ ID NO: 58). or a pharma- ceutically acceptable salt thereof.

[0118] In a further embodiment, the polypeptide of the invention has 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-Cy s]-NH2 (SEQ ID NO: 60), or a pharmaceutical thereof. This includes pharma- ceutically acceptable salts.

[0119] In yet another embodiment, the polypeptide of the invention has the following structural formula: 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-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 This includes pharma- ceutically acceptable salts of the formula:

[0120] In a further embodiment, the polypeptide of the 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 This includes pharma- ceutically acceptable salts of the formula:

[0121] In a further embodiment, the polypeptide of the 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 p-Cys]-NH2 (SEQ ID NO: 66), includes pharma- ceutically acceptable salts thereof.

[0122] In a further embodiment, the polypeptide of the 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 a pharma- ceutical acceptable salt thereof. It is an acceptable salt.

[0123] [Example] (Peptide synthesis) The peptides of the present invention were prepared by conventional solid-phase peptide synthesis. The peptides are coupled to an appropriately selected solid support resin known to be suitable for peptide synthesis. Starting with a C-terminal amino acid derivative that had been ligated, the product was extended in a stepwise fashion. Rink Amide MBHA Resin as a Solid Support for Functionalized Peptide Synthesis For the synthesis of peptides with a free carboxyl function at the C-terminus, a resin (e.g. For example, 2-chlorotrityl chloride resin, Wang resin, or Merrifield resin. ) to form an ester bond with the Fmoc-amino acid. Ester-linked Fmoc-amino acid-resin forms of the formula are commercially available from a variety of sources. , commonly used when feasible.

[0124] (Synthesis of disulfide-cyclized peptides) The linear derivative of the disulfide cyclic peptide amide is Fmoc-conjugated using a solid-phase peptide synthesizer. The Fmoc-Rink amide resin was placed in a reaction vessel and swollen with NMP. It was treated with 20% piperidine in NMP for 15 minutes and then washed three times with NMP. Positive Kaiser test (Kaiser, E., Colescot, R.L., B ossinge, CD & Cook, PI, Anal.Biochem., 199 0, 34:595-598) was used to test the resin. The C-terminus was mixed with Fmoc-amino acid derivatives and HOBt. HBTU reagent and DI The coupling reaction was started by adding PEA. After mixing for 2-3 hours, the reaction A small aliquot of resin was removed from the mixture to determine the end of coupling. The resin was then washed three times with NMP. The entire cycle was repeated using the second C-terminal Fmoc-amino acid derivative as previously described. The same reaction cycle was repeated with each incoming amino acid. The chloranil color test (Vojkovsky, T. Pept. Res. , 1995, 8:236-237) was subjected to Fmoc deprotection from proline residues in the peptide sequence. Test for positive protection and termination of amino acid coupling to proline This was used instead of the Kaiser test to determine the N-terminus (negative chloranil test). For peptides with acetyl groups at the termini, the Fmoc-deprotected peptide resin was chromatographed in acetic anhydride. The resins that were negative in the Kaiser test were treated with acid and pyridine for 10 min. The MP was washed with dichloromethane and dried under reduced pressure. The trifunctional amino acid derivatives used were Fmoc- 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(OBt )-OH.

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

[0126] (Synthesis of lactam cyclized peptides) Cyclic lactam peptides were also synthesized by standard solid-phase peptide synthesis methods. For peptides with Dpr, Fmoc-Dpr(Mtt)-BHA resin was used for solid-phase peptide synthesis. The Fmoc group was removed as described above, and the next Fmoc-protected amine was A monoacid, e.g., Fmoc-Trp(Boc)-OH, is coupled to the aldehyde by standard coupling procedures. The Fmoc protecting group was removed and the carboxyl group was then added to the resin until the amino acid sequence was complete. By repeated coupling and deprotection, the remaining amino acids are individually linked to the correct sequence. For glutamic acid, Fmoc-Glu(OPip) coupling was used. The peptides were then subjected to the same procedure as previously described for the disulfide series. The fully aligned peptides were acetylated at the N-terminus. Then, the orthogonally protected side chains were For example, the orthogonally protected Glu side chain was replaced with 2-phenylisopropyl (O Pip) ester or Dpr as 4-methyltrityl (Mtt) The peptide resin containing the amide 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 dissolved in trifluoroacetate in the presence of water and 1,2-ethanedithiol (EDT). The resin was cleaved with the remaining protecting groups using trifluoroacetic acid (TFA). The product was collected by precipitation and centrifugation. Final purification was performed using reverse phase C The purified peptides were lyophilized using a -18 column and reversed-phase HPLC. and mass analyzed by mass spectrometry using electrospray methodology.

[0127] Examples of compounds of the invention are given in Table 2. Table 2: Examples of compounds of the invention [Table 3] TIFF0007654587000021.tif201149 TIFF0007654587000022.tif173149

[0128] Radioligand binding assay Coupling constant (K d ) or a receptor binding assay to determine the cyclic peptide of the present invention. The inhibitory concentration (IC) for displacing a radiolabeled ligand from its receptor 50 ) to This may be done by any means known in the art.

[0129] As an example, the preparation of cell membranes for binding assays was performed using hMC receptor subtypes 1, 3, The cells were prepared from CHO-K1 cells transfected to stably express 4 or 5. Ru. [125I](Tyr2)-(Nle4-D-Phe7)-α-MSH([125I Competitive inhibition of ]-NDP-a-MSH binding is performed in polypropylene 96-well plates. Briefly, cell membranes (1–10 μg protein / well) prepared as described above were The concentration of the test compound was increased from 0.1 to 0.3 nM [125I]-NDP-α-MSH. 0.2% BSA, 5 mM MgCl2, 1 mM CaCl2 and 0.1 mg / mL bacitracin in 50 mM Tris-HCl, pH 7.4, at 37°C The mixture was incubated for 120 min. The bound [125I]-NDP-α-MSH ligand was , GF / C presoaked in 0.1% (w / v) polyethyleneimine (PEI) Glass fiber plates (Unifilter®, Meriden, CT, The free [125I]-NDP-α-MSH was separated by filtration through a filter (U.S.A.). The filter was washed three times with 50 mM Tris-HCl, pH 7.4, at a temperature of approximately 0-4°C. and then assayed for radioactivity. Binding data are analyzed by regression analysis.

[0130] (Cyclic AMP stimulation assay) Functions for determining the agonist or antagonist status of the cyclic peptides of the present invention The assay may be performed by any means known in the art. (ECL) assay) The stimulation of intracellular cyclic AMP (cAMP) levels by peptides was measured using electrochemiluminescence (ECL) ) assay in a dose-dependent manner (Meso Scale Discov ery, Gaithersburg, MD, USA; hereafter referred to as "MSD"). Briefly, CHO-K1 cells stably expressing hMC receptor subtypes were transformed with RMP I1640® Assay Buffer (0.5 mM IBMX and 0.2% TA Suspended in RMPI1640 buffer containing protein cocktail (MSD blocker A) Approximately 7,000 cells stably expressing hMC receptor subtypes 1, 3, 4 or 5 Transgenic CHO-K1 cells were cultured at 1000 x g / well in a 384-well plate containing integrated carbon electrodes. The plate was then coated with anti-cAMP antibody. Increasing concentrations of test compound are added and the cells are incubated at 37°C for approximately 40 minutes. 0.2% protein cocktail and 2.5 nM TAG TM Ruthenium-labeled Cell lysis buffer (pH 7.3, MgCl2 and Tr) containing cAMP (MSD) A HEPES-buffered saline solution containing 100% ethanol (registered trademark) was added to the cells. The cells are incubated at room temperature for approximately 90 minutes. At the end of the second incubation period, the readout is Capture buffer (pH 7.8 containing ECL coreactants and Triton X-100) The cAMP level in the cell lysate was measured by Sector Immunoassay. Immediate determination was made by ECL detection using the Agilent 6000 Reader (MSD). The data were analyzed by computer-assisted nonlinear regression analysis (X Lfit; IDBS) or reported as EC50 values. EC50 is the maximum response 50% of the response (e.g., the maximal level of cAMP as determined using the assay described above) The concentrations of agonist compounds required to obtain 50% of the target signal are shown.

[0131] (cAMP measurement assay) Human MC4-R transfected cells were cultured at confluence in 96-well plates. The cells were grown to a homogenous state (approximately 250,000 cells / well were seeded). Each was treated with 0.2 mM isobutylmethylxanthine (IBMX) and graded concentrations of peptides. The cells were treated with peptide in the presence of 20 nM NDP-MSH or 20 nM NDP-MSH. Cells similarly treated with SH alone were used as a positive control in a volume of 200 μL. A buffer blank is also included to serve as a negative control. After 1 hour of incubation at 7°C, 50 μL of cell lysis buffer was added. The total c accumulated in the 250 μL of incubation medium is then lysed. AMP was measured using a commercially available low pH cAMP assay kit (Amersham Biosciences). nces) and quantitated according to procedures specified by the kit supplier. The cAMP accumulation was similar to or greater than that of α-MSH as a control. The peptides shown are believed to be agonists. Agonist data was plotted and curved. The line is fitted and the EC50 value is determined. Peptides that show accumulation in the same range as the buffer blank in the presence of ribosomal protein (RIP) are not effective at the concentrations tested. The peptides showing attenuated accumulation were not associated with cAM when α-MSH was also present in the assay. P, which is considered an antagonist. This may also be carried out using C-lR cells, hMC-3R cells and hMC-5R cells.

[0132] (cA using the β-galactosidase (β-Gal) Reporter System) Measurement of MP accumulation) An enzyme flagellar assay using β-galactosidase (β-Gal) as a functional reporter system. A chemiluminescence reading system using the EFC (Enhanced Fragment Complement) system was used. This assay system for the cortin receptor system is commercially available (cAMP Hun ter GPCR Assay System, Discoverx Corp, Fremont, CA). This assay consists of an EA for the enzyme acceptor and an ED for the enzyme donor. This assay utilizes the β-Gal enzyme, which is split into two complementary parts: The ED moiety fused to the MP reacts with cAMP produced by the cell and with an antibody specific to cAMP. EA is then added to compete for binding to unbound ED. This active enzyme then produces an active β-Gal that contains -cAMP. The enzyme converts the signal into an output signal, which is then read by a standard microplate reader. Record it in.

[0133] Briefly, 10,000 cells / well were seeded overnight and then incubated with cell assay buffer. Ink the cells into each well (10 μl of assay buffer) in 5 μL of The test compound and cAMP antibody reagent (5 μL) were incubated with 4-fold concentration gradients. Both are incubated at 37°C for 30 minutes. Primary fragments and reporter substances (Emerald II-Galacton St Add cell lysis buffer (20 μL) containing ar, 5:1, and incubate at room temperature for 60 minutes. Then add 20 μL of EA β-Gal fragment reagent. After 120 min of incubation, the plate was read using a plate reader (Envision). Measure chemiluminescence and use the data to calculate EC50 values ​​for the test peptides.

[0134] The results are shown in Table 3.

[0135] Table 3: EC50 (nM) values ​​of the compounds according to the present invention [Table 4] TIFF0007654587000024.tif168135

[0136] Although the present invention has been particularly shown and described with reference to example embodiments, it will be understood by those skilled in the art that various changes in form and details can be made therein without departing from the scope of the invention as encompassed by the appended claims. In one aspect, the present invention provides the following. [Item 1] An isolated polypeptide having the following structural formula (I): [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R 1 is -H, or C1-C6 acyl; R 2 -NR 3 R 4 -OR 5 and R 3 、R 4 and R 5 are each independently 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 TzAla; or A 1 is a moiety selected from optionally substituted C1-C12 alkyl, optionally substituted C6-C18 aryl, optionally substituted C5-C18 heteroaryl, aralkyl in which the aryl moiety is optionally substituted C6-C18 aryl and the alkyl moiety is optionally substituted C1-C12 alkyl, or heteroaralkyl in which 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 an amino acid residue selected from Cys, hCys, Pen, Asp, Glu, Lys, Orn, Dbu, or Dpr; A 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 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 A 3 does not exist or A 3 is a residue Y, Y having the structural formula

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Claims

1. An isolated polypeptide having the following structural formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, During the ceremony, 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 H or C1-C6 alkyl; A １ is Nle; or A １ is a moiety selected from optionally substituted C1-C12 alkyl, optionally substituted C6-C18 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 １ does not exist; A ２ and A ８ are each independently an amino acid residue selected from Cys, hCys, or Pen; A ２ and A ８ are selected so as to be able to form a covalent bond between each of the side chains in a pair; A ３ is Pro; A ４ does not exist; A ５ is optionally substituted Phe, optionally substituted 1-Nal, or optionally substituted 2-Nal; A ６ is Arg; A ７ is Trp; Polypeptides.

2. A ３ The polypeptide of claim 1, wherein is a D-amino acid.

3. A ５ The polypeptide according to claim 1 or 2, wherein is optionally substituted D-Phe.

4. A ５ At any of the five aromatic carbons, F, Cl, Br, I, -CH ３ , -OH, -CN, amine, -NO ２ , or -OCH ３ The polypeptide according to any one of claims 1 to 3, which may be substituted with a substituent selected from the following:

5. A ５ are 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 ), Phe(4'-OCH ３ ), Phe(4'-CF ３ ), Phe(4'-NO ２ ), Phe (4'-t-Bu), Phe (2',4'-di-F), Phe (2',4'-di-Cl), Phe (2',4'-di-Br), Phe (2',4'-di-I), 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'-diI), Phe(3',5'-di-CN), Phe(3',5'-diCH ３ ), Phe(3',5'-di-OCH ３ 4. The polypeptide of claim 1, wherein the D-amino acid residue is selected from the group consisting of C(3',4',5'-triF), C(3',4',5'-triF), and C(3',4',5'-triF).

6. A composition comprising an effective amount of a polypeptide described in claim 1 or a pharma- ceutically acceptable salt thereof for treating a disorder responsive to the regulation of MC4R in a subject in need of such treatment.

7. The composition of claim 6, wherein the disorder responsive to modulation 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.

8. A pharmaceutical composition comprising the polypeptide of claim 1 or a pharma- ceutically acceptable salt thereof in a pharma- ceutically acceptable carrier.

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