Pharmaceutical composition

Abstract

To provide a pharmaceutical composition that allows extended release of cationic peptide.SOLUTION: A pharmaceutical composition contains an ionic composite containing cationic polypeptide and anionic excipient selected from PEG-carboxylic acid, fatty acid having 10 or more carbon atoms, anionic phospholipid, and a combination thereof.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Patent Application No. 61 / 792,440, filed March 15, 2013. The entire teachings of the above application are hereby incorporated by reference. Be absorbed. [Background technology]

[0002] Enhance the pharmacokinetic properties of an active agent (e.g., to extend the duration of drug action or It is often desirable to minimize any unwanted effects of drugs. In particular, peptide drugs typically dissolve readily in the body, are rapidly absorbed, and are released more slowly. Provides a sudden burst of available drug, as opposed to a gradual release Compositions that can provide a slower or longer release of the drug may be , reduced fluctuation in drug concentration after administration, increased drug usage per dose, in vivo and in vivo Increased in vitro stability and efficacy, reduced toxicity, and less frequent dosing. This can result in increased patient compliance due to the absence of Summary of the Invention [Problem to be solved by the invention]

[0003] Therefore, pharmaceutical compositions containing active agents that provide a long-term release of the active agent are There is a need for a composition. [Means for solving the problem]

[0004] The present invention relates to a method for the preparation of a cationic polypeptide and a PEG-carboxylic acid in various molar ratios, 10 or more. fatty acids having the above carbon atoms, anionic phospholipids, and combinations thereof. The present invention also relates to an ionic complex comprising an anionic excipient. and a pharma- ceutical composition comprising the compound of formula (I) and a pharma- ceutical carrier. The thione polypeptide has pharmacological activity, and the complex, after administration, is a cationic polypeptide alone. Compared to the German, the cationic polypeptide of the complex is more suitable for use as a sustained release formulation, for example. It can provide a desirable pharmacokinetic profile.

[0005] The present invention also provides a method for the preparation of a cationic polypeptide responsive to pharmacological activity of an ionic complex. Ionic complexes and ionic complexes for treating subjects suffering from diseases or disorders The present invention also relates to the use of a pharmaceutical composition comprising the conjugate. In one embodiment, the disorder to be treated is , responsive to modulation of the melanocortin-4 receptor (MC4R) in a subject in need of such treatment - Patents.com The method includes the step of administering an MC4R modification such as that described herein in Formula I. The method comprises administering to a subject an effective amount of an ionic complex containing a cationic polypeptide as a decorating factor. In certain embodiments, the disorder responsive to modulation of MC4R includes type 1 diabetes, type 2 diabetes, Type 2 diabetes, obesity, insulin resistance, metabolic syndrome, male erectile dysfunction, female Disorders, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, alcoholism eating disorder Harm includes substance abuse disorders, cachexia, inflammation, and anxiety.

[0006] In one embodiment, the compounds and compositions of the present invention bind to the melanocortin-1 receptor (MC 1R) to MC4R and melanocortin-3 receptor (MC3R) The compounds and compositions of the present invention have increased effects on blood pressure and therefore have higher selectivity and potency. These include increased appetite, increased heart rate, unwanted effects on sexual arousal, and increased skin pigmentation. In some cases, adverse side effects such as nausea, vomiting, and vomiting may be reduced or eliminated.

[0007] The ionic complexes of the present invention and pharmaceutical compositions containing the ionic complexes are The pharmacokinetic properties of the polypeptide can be enhanced. For example, cationic polypeptides can be The duration of pharmacological action of tidazole is the period from highest to lowest drug exposure in its pharmacokinetic profile. Therefore, the therapeutic dose of the cation exchange The peptides are maintained within a beneficial exposure range in the body, thereby preventing cationic polypeptide drugs from Reduce the potential for unwanted side effects that may result from a single high exposure. Compositions that can provide a slower or longer release of the active agent. The product has the following advantages: less fluctuation in concentration of active agent after administration, increased amount of active agent used per dose, and in vivo and increased in vitro stability and efficacy, reduced toxicity, and ease of administration. This can result in increased patient compliance due to less frequent administration of the drug. The ionic complex composition is administered at least once daily, once weekly, once every two weeks, or once every four weeks. Once every 2 months, once every 3 months, once every 4 months, once every 5 months Effective therapeutic administration across a range of dosages, including once every 6 months or once every 6 months Suitable for. [Brief description of the drawings]

[0008] [Figure 1]FIG. 1 shows the pharmacokinetic profile of each of the identified pharmaceutical compositions following administration to cynomolgus monkeys. [Diagram 2] FIG. 2 shows the pharmacokinetic profile of each of the identified pharmaceutical compositions following administration to cynomolgus monkeys. [Diagram 3] FIG. 3 shows the pharmacokinetic profile of each of the identified pharmaceutical compositions following administration to cynomolgus monkeys. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] A description of exemplary embodiments of the present invention follows.

[0010] Glossary The nomenclature used to define peptides is such that the N-terminal amino group appears on the left and the C- The terminal carboxyl group appears to the right as typically used in the art. .

[0011] As used herein, the term "amino acid" refers to naturally occurring amino acids and Unless otherwise indicated, the amino acids described herein include both natural and unnatural amino acids. All amino acids and their residues found in the compound are either D or L It can be in a three-dimensional configuration.

[0012] Unless otherwise defined, all technical and scientific terms used herein has 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 incorporated by reference in their entirety. No. 6,399,433, filed on Oct. 23, 2003, which are incorporated by reference in their entireties. [Table 1] TIFF2025004067000002.tif212155TIFF2025004067000003.tif35135

[0013] Unless otherwise indicated, all amino acid abbreviations in this disclosure (e.g., Ala) are It refers to an amino acid residue, i.e., the structure -NH-C(R)(R')-(CO)-. , R and R' are each independently hydrogen or the side chain of an amino acid (e.g., For Ala, R=CH3 and R'=H or R and R' are linked to form a ring system. may be formed).

[0014] The designations "Ac" or "NH2" of the polypeptide termini indicate that the corresponding termini are acetylated, respectively. It indicates that the compound is substituted or aminated.

[0015] The phrase "covalent bond between amino acid side chains" refers to a bond between the side chains of the two amino acid residues being discussed. , each of which is meant to contain functional groups capable of forming covalent bonds with each other. Examples of such bonds are disulfide bonds formed by Cys, hCys, or Pen side chains. bridges and other amino acids such as Asp, Glu, Lys, Orn, Dbu, or Dpr. The amino group of one amino acid side chain and the carboxyl group of the other amino acid side chain form a When a covalent bond is formed between amino acid side chains, it is possible to form an amide bond. Such cyclic polypeptides may be represented by the structural formula: or by using the shorthand notation "c()" or "cyclo()". For example, "-c(Cys-Cys)-" or "-cyclo(Cys-Cys)-" has the structure Construction: [ka]

[0016] represents "-c(Asp-Lys)-" or "-cyclo(Asp-Lys)-" refers to the structure: [ka]

[0017] Represents.

[0018] Ionic Complexes: The present invention relates to cationic polypeptides and PEG-carboxylic acids, The anionic excipient is selected from fatty acids having the formula: Ionic Complexes. Cationic Polypeptide vs. Anionic Excipients in Ionic Complexes. The molar ratio can range, for example, from about 1:1 to about 1:10, and the molar ratio can vary depending on the charge of the anionic excipient. The molar ratio is based on the charge of the cationic polypeptide relative to the charge. :3, approx. 1:4, approx. 1:5, approx. 1:6, approx. 1:7, approx. 1:8, approx. 1:9, or 1:10 is often selected from

[0019] Anionic Excipients Anionic phospholipids: As used herein, an anionic phospholipid is a lipid that is a phospholipid or lipids. One or more oxygen atoms of the acid group are deprotonated to form an organophosphate oxoanion and a negatively charged A phospholipid that provides a charged lipid. Naturally occurring anionic phospholipids are typically In fact, C 16 or larger fatty acid chains. Anionic phospholipids have one or more negative charges (e.g. 1, 2, 3, 4, 5, 6, or more). In embodiments, the anionic phospholipid is phosphatidic acid (PA), phosphatidylglycerol (PG), or phosphatidylcholine (PGA). PG, phosphatidylinositol (PI), or phosphatidylserine ( Suitable anionic lipids are selected from L-α-phosphatidic acid, 1-oleoyl phosphatidyl ester (PS). Lysophosphatidic Acid, L-α-Phosphatidylglycerol, 1,2-Di-O-Tetrahydrofuran 1,2-tradecyl-sn-glycero-3-phospho-(1'-rac-glycerol) Dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DM PG), 1,2-dimyristoyl-sn-glycero-3-phospho-L-serine (DMPS ), 1,2-dipalmitoyl-sn-glycero-3-phosphatidic acid (DPPA), 1 ,2-Distearoyl-sn-glycero-3-phosphatidic acid (DSPA), 1,2- Distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), mPEG -2,000-DSPE, mPEG-5,000-DSPE, 1,2-dipalmitoyl- sn-glycero-3-phosphoethanolamine (DPPE), mPEG-2,000-D PPE, mPEG-5,000-DPPE, 1-(1,2-dihexadecanoyl phosphatase 1-(1,tidyl)inositol-4,5-bisphosphate, trisodium salt, and 1-(1, 2-Dihexadecanoylphosphatidyl)inositol-3,4,5-triphosphate , tetrasodium salt, 1-palmitoyl-2-oleoylphosphatidylglycerol (P OPG), 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphoglyceryl rol (PAPG), DSPG (distearoylphosphatidylglycerol), DPP G (dipalmitoyl phosphatidylglycerol), DEPG (dielideyl phosphatidyl tidylglycerol), DOPG (dioleoylphosphatidylglycerol), DEP A (dielaidoylphosphatidic acid), DOPA (dioleoylphosphatidic acid), DSPS (distearoylphosphatidylserine), DPPS (dipalmitoylphospha tidylserine), DEPS (dielaidoylphosphatidylserine), and DOPS ( Dioleoylphosphatidylserine), L-α-Lysophosphatidylserine, L-α-Lysophosphatidylserine Sophosphatidylinositol, Tetradecylphosphonic acid, L-α-phosphatidylinositol L-α-phosphatidylinositol-4,5-bisphosphate Phosphate, 1,2-diphytanoyl-sn-glycero-3-phosphate, 1,2-di- O-tetradecyl-sn-glycero-3-phospho-(1'-rac-glycerol), and mixtures thereof.

[0020] A particular anionic phospholipid for use in the present invention is 1,2-distearoyl-s n-Glycerol-3-phosphoethanolamine (DSPE) conjugated polyethylene Recall that this structure is as follows: [ka]

[0021] The value of "n" varies depending on the molecular weight. Such anionic phospholipids are referred to herein as In this case, it is called mPEG-(Mol. Wt.)DSPE. mPEG-5,000-DSPE and mPEG-5,000-DSPE, Another example is mP EG-2,000-DPPE and mPEG-5,000-DPPE, DPPE is , 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine.

[0022] C 16 Synthetic anionic phospholipids having chain lengths shorter than 1,2-diaminodiphenylphosphorylamide (DPPH) are also available. Synthetic lipids can also be obtained by adding fatty acids that provide additional anionic groups in the fatty acid chain. For example, it is possible to incorporate shorter fatty acid chains terminating in a carboxylate group. -Palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine.

[0023] These naturally occurring and synthetic anionic lipids are Avanti polar lipids. pids (Alabaster, AL) or Lipoid LLC (Newark, N J) or Cordon Pharma (Boulder, CO) or NOF Cor Some commercially available products, such as those from America (White Plains, NY), may be obtained from a source.

[0024] fatty acid: As used herein, a "fatty acid" is a carboxylate having a long aliphatic tail (chain). A carboxylic acid is either saturated or unsaturated and has 10 or more carbon atoms. The carbon atom chain may be linear, branched, saturated, monounsaturated, or polyunsaturated. It is possible.

[0025] Suitable saturated fatty acids are capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, These include phosphoric acid, arachidic acid, docosanoic acid, tetracosanoic acid, and hexacosanoic acid, Not limited to these.

[0026] Suitable unsaturated fatty acids are cis-2-decenoic acid, myristoleic acid, palmitoleic acid. , sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid , α-linolenic acid, arachidonic acid, cis-parinaric acid, eicosapentaenoic acid, and Including but not limited to phytanic acid.

[0027] PEG-Carboxylic Acid: As used herein, "PEG-carboxylic acid" refers to a carboxylic acid functionalized Polyethylene glycol (PEG) polymers are polymers that have the ability to be polymerized with a single functional group. (monocarboxylate) or homobifunctional (dicarboxylate) Functionalized PEG can be linear or branched. The PEG-carboxylic acid suitable for use in the present invention is about 1,000 to about 100 The copolymer may have an average molecular weight range of up to 10,000.

[0028] Homobifunctional PEGs suitable for use in the present invention are generally COOH-PEG-COOH As OH or chemical formula: COOH-(CH2CH2O) n -CH2CH2-COOH Or COOH-CH2CH2-CO-O(CH2CH2O) n -CH2CH2-COO H or COOH-CH2CH2-CO-O(CH2CH2O) n -CO-CH2CH 2-COOH, where the value of "n" varies depending on the molecular weight. For example, PEG-10,000-dicarboxylate, shown in the examples. The carboxylic acid is a homobifunctional PEG with a molecular weight of 10,000. An average molecular weight range of 1.0 to about 100,000 is suitable. Typically, the molecular weight is The range can range from 00 to 40,000.

[0029] One type of monofunctional PEG-carboxylic acid suitable for use in the present invention is generally PEG-COOH or formula: CH3O-(CH2CH2O) n -CH2CH2 -COOH or CH3O-(CH2CH2O) n -CO-CH2CH2-COOH and where the value of "n" varies with molecular weight. For example, The mPEG-10000 used in the examples has a methoxy group, as shown in the formula: The average molecular weight range of about 1,000 to about 100,000 is Typically, molecular weights range from 1,000 to 40,000. can be done.

[0030] Another type of monofunctional PEG-carboxylic acid suitable for use in the present invention is generally PEG-COOH or with the formula: HO-(CH2CH2O) n -CH2CH2 -COOH, where the value of "n" varies depending on the molecular weight. For example, PEG-10,000-monocarboxylate, shown in the examples The carboxylic acid is a monofunctional PEG having a molecular weight of 10,000. Average molecular weight ranges of 100,000 are suitable. Typically, molecular weights are between 1,000 and 1,000. It can range from ~40,000.

[0031] Suitable PEG-carboxylic acids include PEG-10,000-monocarboxylate, PEG -20,000-monocarboxylate, mPEG-1,000-monocarboxylate , mPEG-2,000-monocarboxylate, mPEG-5,000-monocarboxylate silyl, mPEG-10,000-monocarboxylate, mPEG-20,000- Monocarboxylate, mPEG-30,000-monocarboxylate, mPEG-4 0,000-monocarboxylate, PEG-1,000-dicarboxylate, PEG -2,000-dicarboxylate, PEG-3,500-dicarboxylate, PEG -5,000-dicarboxylate, PEG-7,500-dicarboxylate, PEG -10,000-dicarboxylate, and Y-shaped PEG-40,000-monocarboxylate The above-mentioned Y-shaped PEG-carboxylic acids include, but are not limited to, silylates. can be shown as follows: [ka]

[0032] Furthermore, the value of "n" determines the molecular weight.

[0033] In certain embodiments, the PEG-carboxylic acid is PEG-5,000-monocarboxylate. PEG-10,000-monocarboxylate, PEG-20,000-monocarboxylate carboxylate, mPEG-5,000-monocarboxylate, mPEG-10,000 -monocarboxylate, mPEG-20,000-monocarboxylate, PEG-5 ,000-dicarboxylate, and PEG 10,000-dicarboxylate. It is selected.

[0034] Suitable PEG-carboxylic acids are available from Nanocs, Inc. (New York, NY). , JenKem Technology (Allen, TX), or NOF Corp It is commercially available from America (White Plains, NY).

[0035] In certain embodiments, two or more anionic excipients (e.g., PEG- Carboxylic acids, anionic phospholipids, and fatty acids) can be used in the formulation. For example, PEG-carboxylic acids and anionic phospholipids, PEG-carboxylic acids and lipids Acids, or fatty acids and anionic phospholipids, may be used in the pharmaceutical formulations of the present invention. In another embodiment, the phospholipid is made of a fatty acid, an anionic phospholipid, and a PEG-carboxylic acid. Three or more selected anionic excipients are present in the pharmaceutical formulation. In certain embodiments, The combination of anionic excipients was stearic acid and mPEG-2,000-DSPE, DPPA and PEG-10,000-dicarboxylate, DPPA and PEG-3 ,350, DPPA, DPPA and mPEG-3,350 and mPEG-2,000 In another embodiment, the anionic ligand described herein is selected from -DSPE. The combination of mPEG-2000-DSPE and CMC was also used. In addition, it can be further combined with carboxymethylcellulose (CMC).

[0036] Cationic peptides: As used herein, a "cationic polypeptide" is a polypeptide that has a pH of about 5.0. It means any polypeptide that carries a positive charge. Cationic polypeptides are naturally occurring It may comprise amino acid residues, non-naturally occurring amino acid residues, or a mixture thereof. The positive charge of a cationic polypeptide is located in the side chain of an amino acid in the polypeptide sequence. Cations present as part of the alpha amino group of an amino acid in the The positive charge of cationic polypeptides can also occur at the termini of the peptide sequence or at the Cationic functional groups ligated to the polypeptide at the side chains of the amino acids in the sequence There may be more than one cationic functional group present in a cationic polypeptide. For example, cationic polypeptides may contain 1, 2, 3, 4, 5, 6, or more cationic polypeptides. Each of these can provide a positive charge. Such functional groups include, for example, amino groups (primary, secondary, or tertiary), quaternary Ammonium group, guanidino group, amidino group, pyridine group, imidazole group, phosphonium In certain embodiments, the cationic group includes an amino group, a glycol group, and a sulfonium group. It is an anidino group or an imidazole group.

[0037] The cationic polypeptides of the present invention have one or more chiral centers and therefore many All stereoisomers and mixtures thereof are included within the scope of the present invention. The racemate is separated using preparative HPLC and a column with a chiral stationary phase. The individual enantiomers may be separated or isolated using methods known to those skilled in the art. Moreover, the chiral intermediate compounds may be separated and used to produce the chiral intermediate compounds of the present invention. It may be used to prepare chiral compounds.

[0038] The cationic polypeptides described herein may exist in one or more tautomeric forms. All tautomers and mixtures thereof are included within the scope of the present invention.

[0039] Cationic polypeptides suitable for use in the present invention include polypeptides (polypeptides eptides categories) and the specific polypeptides and and the polypeptides represented by Formula I. The peptides are pharmacologically active.

[0040] LHRH (GnRH) agonists: Leuprolide: Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Ar g-Pro-NHEt (SEQ ID NO: 46) Buserelin: Pyr-His-Trp-Ser-Tyr-D-Ser(tBu)-Leu -Arg-Pro-NHEt (SEQ ID NO: 47): Histrelin: Pyr-His-Trp-Ser-Tyr-D-His(Bzl)-Le u-Arg-Pro-NHEt (SEQ ID NO: 48) Goserelin: Pyr-His-Trp-Ser-Tyr-D-Ser(tBu)-Leu -Arg-Pro-Azagly-NH2 (SEQ ID NO: 49) Deslorelin Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg -Pro-NHEt (SEQ ID NO: 50) Nafarelin Pyr-His-Trp-Ser-Tyr-D-Nal(2)-Leu- Arg-Pro-Gly-NH2 (SEQ ID NO: 51) Triptorelin Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Ar g-Pro-Gly-NH2 (SEQ ID NO: 52) Aborelin Pyr-His-Trp-Ser-Tyr-D-Trp(2-Me)-Le u-Arg-Pro-NHEt (SEQ ID NO: 53) GnRH antagonists: Abarelix: Ac-D-Nal(2)-D-Phe(4Cl)-D-Pal(3)- Ser-(Me)Tyr-D-Asn-Leu- Lys(iPr)-Pro-D-Al a-NH2 (SEQ ID NO: 54) Cetrorelix: Ac-D-Nal(2)-D-Phe(4Cl)-D-Pal(3)- Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2 (sequence no. No.:55) Degarelix: Ac-D-Nal(2)-D-Phe(4Cl)-Pal(3)-Ser -Phe(4-(4S)-hexahydro-2,6-dioxo-4-pyrimidinyl(carbo (4-guanidino)-D-Phe(4-amino)-Leu-Lys(iPr)-Pro- D-AlaNH2 (SEQ ID NO: 56) Ganirelix: Ac-D-Nal(2)-D-Cpa(4)-D-Pal(3)-Ser -Tyr-D-Harg(Et)2-Leu- Harg(Et)2-Pro-D-Al a-NH2 (SEQ ID NO: 57) Somatostatin analogues: Octreotide D-Phe-[Cys-Phe-D-Trp-Lys-Thr-Cys ]-Thr-ol (SEQ ID NO:58) Lanreotide D-Nal(2)-[Cys-Tyr-D-Trp-Lys-Val-C ys]-Thr-NH2 (SEQ ID NO: 59) Vapreotide D-Phe-[Cys-Tyr-D-Trp-Lys-Val-Cys] -Trp-NH2 (SEQ ID NO: 60) Pasireotide (Signifor) Cyclo[-(4R)-4-(2-aminoethyl carbamazepine] Bamoyloxy)-L-prolyl-L-phenylglycyl-D-tryptophyl-L-lysine Cyl-4-O-benzyl-L-tyrosyl-L-phenylalanyl-] (SEQ ID NO: 60) Other peptides: Glucagon Amylin Pramlintide Insulin Glucagon-like peptide-1 (GLP-1) GLP-1 agonists Exenatide Parathyroid hormone (PTH) Adrenocorticotropic hormone (ACTH) Botulinum toxin Amyloid peptides Cholecystokinin Calcitonin Conotoxins Prialt Gastric inhibitory peptide Insulin-like growth factors, including recombinantly produced IGF-1, such as Increlex child Growth hormone-releasing factor antibacterial factors Glatiramer (Copaxone) Hematide (peginesatide) Nesiritide ANF ​​Peptide Angiotensin Peptides ACTH Human Growth Hormone (hGH), including recombinantly produced hGH Melanocortin Opioid peptides Dynorphin Oxytocin Oxytocin analogs, including antagonists Vasopressin and analogs, and Somatostatin and its analogs Cationic polypeptides of formula I: The cationic polypeptide for use in the present invention is a cationic polypeptide of formula (I) or a pharma- ceutically acceptable salt thereof [ka]

[0041] or a pharma- ceutically acceptable salt thereof; 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, respectively, is independently 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 a, or A 1 is an optionally substituted C1-C12 alkyl, an optionally substituted C6-C1 8 aryl, optionally substituted C5 to C18 heteroaryl, and aralkyl. The aryl portion is an optionally substituted C6 to C18 aryl, The alkyl portion is an optionally substituted C1-C12 alkyl or heteroaralkyl. and the heteroaryl portion is an optionally substituted C5-C18 heteroaryl. and the alkyl portion is an optionally substituted C1-C12 alkyl. 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 pairwise selected to be able to form covalent bonds between their respective side chains. And, A 3 is absent or Ala, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, or Aib, A 4 is absent or Atc, Ala, QAla, Aib, Sar, Ser, Thr , Pro, Hyp, Asn, Gln, optionally substituted His, Trp, Tyr, L an amino acid residue selected from the group consisting of ys, Arg, sChp, or residue X, wherein X is An amino acid represented by the structural formula: [ka]

[0042] A 5 is optionally substituted Phe, optionally substituted 1-Nal, or 2-Nal, which may be A 6 is Arg, A 7 is Trp, All amino acid residues may be in either the L- or D-configuration.

[0043] In an exemplary embodiment, A 3 and A 4 But both are not absent. The values ​​and preferred values ​​of the variables are as defined above and below for formula (I). do.

[0044] In an exemplary embodiment, A 4 If is an amino acid, A 3 It is neither Aib nor Gly. Values ​​and preferred values ​​for the remaining variables are as defined above for formula (I). do.

[0045] 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 values ​​and preferred The values ​​are as defined above for formula (I).

[0046] In an exemplary embodiment, each A 2 and A 8 Cys, hCys, or Pe n, (a) A 4 If absent, A 3 is not L-His, ( b) A 3 If absent, A 4 is not L-His, (c)A 4 If is His, If, A 3 is not Glu, Leu, or Lys. The values ​​of the remaining variables and the preferred The values ​​are as defined above for formula (I).

[0047] In an exemplary embodiment, 1) A 3 and A 4 However, both are not absent. )A 4 If is an amino acid, A 3 is neither Aib nor Gly, 3) A 4 His Yes, A 5 is D-Phe or 2-Nal, A 3 D-amino acids and LA Not la, 4) each A 2 and A 8 from Cys, hCys, or Pen If selected, (a) A 4 If absent, A 3 is not L-His, (b) A 3 If absent, A 4 is not L-His, (c)A 4 If is His, then A 3 is not Glu, Leu, or Lys. The values ​​and preferred values ​​of the remaining variables are: As defined above for formula (I).

[0048] In another embodiment, in the polypeptide of formula (I), A 4 are L-amino acids. In terms of form, A 4 is absent. Values ​​and preferred values ​​of the remaining variables are as follows for formula (I): As defined above.

[0049] In an exemplary embodiment, A 5 is optionally substituted 1-Nal or It can be optionally substituted 2-Nal, for example optionally substituted D-2-Nal. . A 5 can be substituted at any of the five aromatic carbons, and the substituents are F, CI, B r, I, -CH3, -OH, -CN, amine, -NO2, or -OCH3 do.

[0050] In a further embodiment, in the polypeptide of formula (I), A 5 is an optionally substituted D- Phe. 5 can be substituted at any of the five aromatic carbons, and the substituents are F, CI, Br, I, -CH3, -OH, -CN, amine, -NO2, or -OCH3 Selected from: A 5 Suitable examples are 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'-NO 2), 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'-CN), Ph e(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), Ph e(2',4'-di-CN), Phe(2',4'-di-CH3), Phe(2',4' -di-OCH3), Phe(3',4'-diF), Phe(3',4'-diCl), Ph e(3',4'-di-Br), Phe(3',4'-di-l), Phe(3',4'-di-C N), Phe(3',4'-di-CH3), Phe(3',4'-di-OCH3), Ph e(3',5'-diF), Phe(3',5'-diCl), Phe(3',5'-diBr ), Phe(3',5'-di-l), Phe(3', 5'-di-CN), Phe(3', 5'-diCH3), Phe(3',5'-di-OCH3), or Phe(3',4', 5'-triF) and D-amino acid residues selected from the group consisting of, but not limited to, the following: The values ​​and preferred values ​​of the variables are as defined above for formula (I).

[0051] In a further embodiment, in the polypeptide of formula (I), A 5 is an optionally substituted D- 2-Nal. A 5 can be substituted at any of the five aromatic carbons, is F, CI, Br, I, -CH3, -OH, -CN, amine, -NO2, or -OC Selected from H3.

[0052] In another embodiment, in the polypeptide of formula (I), A 4 However, any substitutable and His optionally substituted at the substituted (cancellable) position, the substituents being F, CI, B r, I, -CH3, -OH, -CN, amine, -NO2, benzyl, or -OCH3 Values ​​and preferred values ​​for the remaining variables are defined above for formula (I). That is correct.

[0053] In certain embodiments, compounds of the invention have an EC 50 (MC1R) / E C50 (MC4R), about 0.01 nM to about 10 nM, for example 0.01 EC for MC4R of ~3 nM 50 Another embodiment is a polypeptide of formula (I) having the formula: In one embodiment, the polypeptide of the present invention is a polypeptide represented by any of the following structural formulas: Chid: [ka] TIFF2025004067000011.tif204165

[0054] or a pharma- ceutically acceptable salt thereof.

[0055] In other embodiments, a polypeptide of the invention has any one of the following structural formulas: [ka]

[0056] or a pharma- ceutically acceptable salt thereof.

[0057] In further embodiments, the polypeptide of the invention is represented by any one of the following structural formulas: A polypeptide represented by: [ka]

[0058] or a pharma- ceutically acceptable salt thereof.

[0059] In another embodiment, the polypeptide of the present invention is a polypeptide represented by formula (I): Including A 4 But, Atc, Ala, QAla, Aib, Sar, Ser, Thr, Pro , Hyp, Asn, Gln, substitutions His, Trp, Tyr, Lys, Arg, sChp, or residue X. An example of such a peptide is A peptide represented by any one of the formulas: Ac-Arg-Cyclo[Cys-D-Ala-His(3-Me)-D-Phe-Arg -Trp-Cys]-NH2 (SEQ ID NO:6), Ac-Arg-Cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Arg -Trp-Cys]-NH2 (SEQ ID NO: 7), Ac-Arg-cyclo[Cys-D-Ala-Trp-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 8), Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 9), Ac-Arg-Cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 10), Ac-Arg-cyclo[Cys-D-Ala-Arg-D-Phe-Arg-Trp-C ys]-NH (SEQ ID NO: 11), Ac-Arg-Cyclo[Cys-D-Ala-Tyr-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 12), Ac-Arg-cyclo[Cys-D-Ala-D-Pro-D-Phe-Arg-Trp -Cys]-NH2 (SEQ ID NO: 13), Ac-Arg-Cyclo[Cys-D-Ala-Pro-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 14), Ac-Arg-Cyclo[Cys-D-Ala-Pro-D-Phe(pF)-Arg- Trp-Cys]-NH2 (SEQ ID NO: 15), Ac-Arg-Cyclo[Cys-D-Ala-Atc-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 16), Ac-Arg-cyclo[Cys-D-Ala-QAla-D-Phe-Arg-Trp- Cys]-NH2 (SEQ ID NO: 17), Ac-Arg-Cyclo[Cys-D-Ala-sChp-D-Phe-Arg-Trp- Cys]-NH2 (SEQ ID NO: 18), or Ac-Arg-Cyclo[Cys-D-Ala-XD-Phe-Arg-Trp-Cys ]-NH2 (SEQ ID NO: 19) or This includes pharma- ceutically acceptable salts thereof.

[0060] In an exemplary embodiment, the polypeptide of the invention has any one of the following structural formulas: A polypeptide represented by: Ac-Arg-Cyclo[hCys-Ala-D-Phe-Arg-Trp-Cys]-N H2 (SEQ ID NO: 20), Ac-Arg-Cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Cys] -NH2 (SEQ ID NO: 21), Ac-Arg-cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Pen] -NH2 (SEQ ID NO: 22), Ac-Arg-cyclo[Glu-D-Ala-D-Phe-Arg-Trp-Dpr]- NH2 (SEQ ID NO: 23), Ac-Arg-cyclo[Glu-Ala-D-Phe-Arg-Trp-Dpr]-NH 2 (SEQ ID NO: 24), Ac-Arg-Cyclo[hCys-Aib-D-Phe-Arg-Trp-Cys]-N H2 (SEQ ID NO: 25), Ac-Arg-cyclo[hCys-Sar-D-Phe-Arg-Trp-Cys]-N H2 (SEQ ID NO: 26), Ac-Arg-Cyclo[hCys-Val-D-Phe-Arg-Trp-Cys]-N H2 (SEQ ID NO: 27), Ac-Arg-cyclo[hCys-D-Val-D-Phe-Arg-Trp-Cys] -NH2 (SEQ ID NO: 28), Ac-Arg-cyclo[hCys-Gln-D-Phe-Arg-Trp-Cys]-N H2 (SEQ ID NO: 29), Ac-Arg-cyclo[hCys-D-Gln-D-Phe-Arg-Trp-Cys] -NH2 (SEQ ID NO: 30), Ac-Arg-cyclo[hCys-Ala-D-Phe-Arg-Trp-Pen]-N H2 (SEQ ID NO: 31), Ac-Arg-cyclo[D-Pen-D-Ala-D-Phe-Arg-Trp-hCy s]-NH2 (SEQ ID NO: 32), Ac-Arg-cyclo[Cys-D-Ala-D-Phe-Arg-Trp-hCys] -NH2 (SEQ ID NO: 33), Ac-Arg-cyclo[Pen-D-Ala-D-Phe-Arg-Trp-hCys] -NH2 (SEQ ID NO: 34), Ac-Arg-cyclo[D-hCys-D-Ala-D-Phe-Arg-Trp-Cy s]-NH2 (SEQ ID NO: 35), Ac-Arg-cyclo[hCys-Pro-D-Phe-Arg-Trp-Cys]-N H2 (SEQ ID NO:36), or Ac-Arg-cyclo[hCys-D-Pro-D-Phe-Arg-Trp-Cys] -NH2 (SEQ ID NO: 37) or a pharma- ceutically acceptable salt thereof.

[0061] In another embodiment, the polypeptide of the present invention is a polypeptide represented by formula (I): Including A 3 But, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, From Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, or Aib is a selected amino acid residue, 4 But, Atc, Ala, QAla, Aib, Sar, Ser, Thr, Pro, Hyp, Asn, Gln, substituted His, Trp, Tyr, Ly s, Arg, sChp, or residue X. An example of a polypeptide is a polypeptide represented by any one of the following structural formulas: Ac-Arg-Cyclo[Cys-Val-Gln-D-Phe-Arg-Trp-Cys ]-NH2 (SEQ ID NO: 38), Ac-Arg-Cyclo[Cys-D-Val-Gln-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO:39), or Ac-Arg-Cyclo[Cys-D-Val-His(1-Me)-D-Phe-Arg -Trp-Cys]-NH2 (SEQ ID NO: 40) or a pharma- ceutically acceptable salt thereof.

[0062] In further embodiments, the polypeptide of the invention is represented by any one of the following structural formulas: A polypeptide represented by: Ac-TzAla-cyclo[Cys-Ala-Gln-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 41) or Ac-Glu-Cyclo[Cys-Ala-His-D-Phe-Arg-Trp-Cys ]-NH2 (SEQ ID NO: 42) or a pharma- ceutically acceptable salt thereof.

[0063] In another embodiment, the polypeptide of the invention is represented by any one of the following structural formulas: Polypeptides to be synthesized: Ac-Arg-Cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Arg -Trp-Cys]-NH2 (SEQ ID NO: 7) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 9) Ac-Arg-Cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 10), or a pharma- ceutically acceptable salt thereof.

[0064] In further embodiments, the polypeptide of the invention is represented by any one of the following structural formulas: A polypeptide represented by: Ac-Arg-Cyclo[Cys-D-Leu-His-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO:2), Ac-Arg-Cyclo[Cys-D-Ile-His-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO:3), Ac-Arg-Cyclo[Cys-D-Tle-His-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO: 4), Ac-Arg-Cyclo[Cys-D-Val-His-D-Phe-Arg-Trp-C ys]-NH2 (SEQ ID NO:5), or a pharma- ceutically acceptable salt thereof.

[0065] In further embodiments, the polypeptide of the invention is represented by any one of the following structural formulas: A polypeptide represented by: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-2-Nal-A rg-Trp-Cys]-NH2 (SEQ ID NO: 84), Ac-Arg-cyclo[Cys-D-Ala-Gln-D-2-Nal-Arg-Trp -Cys]-NH2 (SEQ ID NO: 85), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-2-Nal-Arg-Trp -Cys]-NH2 (SEQ ID NO: 86) or a pharma- ceutically acceptable salt thereof. Furthermore, the peptide of the present invention has the following structural formula: A polypeptide represented by any one of: Ac-Arg-Cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Arg -Trp-Cys]-OH (SEQ ID NO: 87), Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp-C ys]-OH (SEQ ID NO:88), or Ac-Arg-Cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp-C ys]-OH (SEQ ID NO: 89) or a pharma- ceutically acceptable salt thereof.

[0066] In another embodiment, the cationic peptide for use in the present invention has the formula (II): It is a thione peptide or a pharma- ceutically acceptable salt thereof.

[0067] An isolated polypeptide having the following structural formula (II): [ka]

[0068] or a pharma- ceutically acceptable salt thereof, 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, respectively, is independently H or C1-C6 alkyl; A 1 is absent or A 1 are Arg, Lys, Orn, His, Nle, Phe, Val, Leu, Trp, Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzAl a, or A 1 is an optionally substituted C1-C12 alkyl, an optionally substituted C6-C1 8 aryl, optionally substituted C5 to C18 heteroaryl, and aralkyl. The aryl portion is an optionally substituted C6 to C18 aryl, The alkyl portion is an optionally substituted C1-C12 alkyl or heteroaralkyl. and the heteroaryl portion is an optionally substituted C5-C18 heteroaryl. and the alkyl portion is an optionally substituted C1-C12 alkyl. 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 pairwise selected to be able to form covalent bonds between their respective side chains. And, A 3 is absent or Ala, Tle, Val, Leu, Ile, Cha, Pro, Ser, Thr, Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, Aib, or a residue Y, where Y is an amino acid residue selected from the group consisting of and wherein the amino acid is selected from the group consisting of: [ka]

[0069] R 11 and R 12 are each independently H, -CH3, phenyl, or benzyl and R 21 , R 22 , R 23 , and R 24 are each independently H, -CH3, or -CF 3, phenyl, benzyl, F, CI, Br, I, -OCH3, or -OH; R 31 , R 32 , R 33 , R 34 , R 41 , R42 , and R 43 are independent of each other. H, -CH3, -CF3, phenyl, benzyl, F, CI, Br, I, -OCH3, or -OH, A 4 is absent or Atc, Ala, QAla, Aib, Sar, Ser, Thr , Pro, Hyp, Asn, Gln, optionally substituted His, Trp, Tyr, L an amino acid residue selected from the group consisting of ys, Arg, sChp, or residue X, wherein X is An amino acid represented by the structural formula: [ka] TIFF2025004067000017.tif60135

[0070] 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 -CF 3, phenyl, benzyl, F, CI, Br, I, -OCH3, or -OH; R 71 , R 72 , R 73 , R 74 , R 8 1. R 82 , and R 83 are independent of each other. H, -CH3, -CF3, phenyl, benzyl, F, CI, Br, I, -OCH3, or -OH, A 5 is optionally substituted Phe, optionally substituted 1-Nal, or 2-Nal, which may be A6 is Arg, A 7 is Trp, All amino acid residues may be in either the L- or D-configuration.

[0071] In an exemplary embodiment, A 3 and A 4 are each independently represented by the following structural formula: The amino acid residue is selected from the amino acids shown below. [ka]

[0072] Values ​​and preferred values ​​for the remaining variables are as defined herein for Formula (I). It is.

[0073] Alone or with "hydroxyalkyl", "alkoxyalkyl", "alkylamine" "Alkyl" used as part of a larger component such as It refers to a linear or branched saturated aliphatic group having, typically, 1 to 12 carbon atoms. In particular, the aliphatic group may have 1 to 8, 1 to 6, or 1 to 4 carbon atoms. The term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t Illustrated by groups such as ethyl-butyl, n-hexyl, and the like. can be.

[0074] "Haloalkyl" refers to an alkyl group substituted with one or more halogen atoms.

[0075] "Halogen" and "halo" refer to fluoro, chloro, bromo, or iodo.

[0076] "Cyano" refers to the group --CN.

[0077] "Ph" refers to a phenyl group.

[0078] "Carbonyl" refers to the divalent group -C(O)-.

[0079] "Aralkyl" used alone or as part of a larger component, as in "aralkyl" "Role" refers to an aromatic carbocyclic group having 6 to 18 carbon atoms with a single ring or multiple condensed rings. The term "aryl" also refers to a group fused to a cycloalkyl or heterocycloalkyl group. Examples of aryl groups include phenyl, Benzo[d][1,3]dioxole, naphthyl, phenanthrenyl, and others Includes those listed below.

[0080] "Aryloxy" refers to the group -OAr, where O is an oxygen atom and Ar is as defined above. is an aryl group as defined above.

[0081] "Aralkyl" includes benzyl, -(CH2)2phenyl, -(CH2)3phenyl, - Replaced with an aryl moiety, such as CH(phenyl)2, and the like It refers to an alkyl having at least one alkyl hydrogen atom.

[0082] Used alone or as part of a larger moiety, such as in "heteroaralkyl" "Heteroaryl" refers to an aryl group consisting of 1 to 4 ring heteroaryls independently selected from nitrogen, oxygen, and sulfur. A 5- to 18-membered monocyclic, bicyclic, or tricyclic heteroaromatic ring system containing a cyclic atom. The term "heteroaryl" also refers to a group fused to a cycloalkyl or heterocycloalkyl group. Particular examples of heteroaryl groups include 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 1,3,4-triazinyl, 1,2,3-triazinyl, benzofuryl, [2,3- Dihydro]benzofuryl, isobenzofuryl, benzothienyl, benzotriazolyl, isobenzofuryl Sobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzimidazolyl aryl, imidazo[1,2-a]pyridyl, benzothiazolyl, benzoxazolyl, Nolidinyl, quinazolinyl, phthalazinyl, quinoxalinyl , cinnolinyl, napthyridinyl, pyrido[3,4-b ]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4,3-b]pyridyl, quinolyl , isoquinolyl, tetrazolyl, 1,2,3,4-tetrahydroquinolyl, 1,2,3, 4-Tetrahydroisoquinolyl, purinyl, pteridinyl, carbazolyl, xanthenyl , benzoquinolyl, and the like.

[0083] "Heteroaralkyl" includes -CH2-pyridinyl, -CH2-pyrimidinyl, and the like. At least one alkyl group replaced with a heteroaryl moiety such as It refers to an alkyl group having a hydrogen atom.

[0084] "Alkoxy" refers to the group -OR, where R is an "alkyl", "cycloalkyl", Examples of alkoxy groups are, for example, methoxy, Includes ethoxy, etheneoxy, and the like.

[0085] "Hydroxyalkyl" and "alkoxyalkyl" refer to hydroxyl and alkoxyalkyl, respectively. and alkyl groups substituted with alkoxy.

[0086] "Amino" means -NH2, "alkylamine" and "dialkylamine" are and -NHR and -NR2, respectively, where R is an alkyl group. "Dicycloalkylamine" and "dicycloalkylamine" are compounds in which R is a cycloalkyl group. "Cycloalkylalkylamine" refers to an amine in which R is a cycloalkyl group. "Cycloalkylalkyl][Alkyl] is -NHR. "Cycloalkylamine" means a cycloalkylamine in which one R is cycloalkyl and the other R is alkyl. It means -N(R)2.

[0087] "Acyl" refers to R"-C(O), where R" is H, alkyl, substituted alkyl, or heteroaryl. Alkyl, substituted heteroalkyl, alkenyl, substituted alkenyl, aryl, alkylaryl or substituted alkylaryl, and in the general formula of certain embodiments, " is shown.

[0088] Suitable substituents for "alkyl", "aryl", or "heteroaryl" are are substituents that will result in the formation of stable compounds of the invention. Examples of suitable substituents are halo, Gen, -CN, -OH, -NH2, (C1-C4) alkyl, (C1-C4) haloalkyl Aryl, aryl, heteroaryl, (C3-C7)cycloalkyl, (5-7 membered)heteroaryl Chloroalkyl, -NH(C1-C6)alkyl, -N((C1-C6)alkyl)2, ( C1-C6) alkoxy, (C1-C6) alkoxycarbonyl, -CONH2, -OC ONH2, -NHCONH2, -N(C1-C6) alkylCONH2, -N(C1-C 6) AlkylCONH(C1-C6)alkyl, -NHCONH(C1-C6)alkyl , -NHCON((C1-C6) alkyl)2, -N(C1-C6) alkylCON(( C1-C6)alkyl)2, -NHC(S)NH2, -N(C1-C6)alkylC(S )NH2, -N(C1~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(C 1~C6) alkyl, -OCONH(C1~C6) alkyl-CON((C1~C6) a alkyl), -C(S)(C1-C6) alkyl, -S(O) p (C1-C6) Alkyl , -S(O) p NH2, -S(O) p NH(C1-C6) alkyl, -S(O)pN(( C1-C6) alkyl)2, -CO(C1-C6) alkyl, -OCO(C1-C6) a alkyl, -C(O)O(C1-C6) alkyl, -OC(O)O(C1-C6) alkyl , -C(O)H, or -COH. The substituents are halogen, -CN, -OH, -NH2, (C1-C4) alkyl, (C1 (C1-C4) haloalkyl, (C1-C4) alkoxy, phenyl, and (C3-C7) cycloalkyl. Within the framework of the present invention, the term "substitution" also refers to the substitution of a hydrogen atom with a heavy atom. It is also intended to encompass the situation where p is an integer having a value of 1 or 2, and p is replaced with a hydrogen atom. do.

[0089] Suitable substituents on the substituted Phe include 1 to 5 substituents on any aromatic carbon, is F, CI, Br, I, -CH3, -OH, -CN, amine, -NO2, or -OC H3. Examples include 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), P he(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'-CN), Phe(4'-C H3), Phe(4'-OCH3), Phe(4'-CF3), Phe(4'-NO2) , Phe(4'-t-Bu), Phe(2',4'-diF), Phe(2',4'-diC l), Phe(2',4'-diBr), Phe(2',4'-dil), Phe(2',4 '-di-CN), Phe(2',4'-di-CH3), 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-CH3), Phe(3',4'-di-OCH3), Phe(3',5 '-diF), Phe(3',5'-diCl), Phe(3',5'-diBr), Phe( 3',5'-di-l), Phe(3',5'-di-CN), Phe(3',5'-di-CH 3), Phe(3',5'-di-OCH3), or Phe(3',4',5'-triF ).

[0090] Suitable substituents on a substituted His include 1 to 3 substituents on any substitutable ring atom. The substituents are F, CI, Br, I, -CH3, -OH, -CN, amine, -NO2, benzene. Examples are 1-methyl-histidine and 3-methyl-histidine. Contains tyl-histidine.

[0091] Name "(amino acid) n " means that an amino acid is repeated n times. For example, The names "(Pro)2" and "(Arg)3" refer to the proline and arginine residues that This means that each is repeated two or three times.

[0092] Pharmaceutically acceptable salts of the compounds disclosed herein are included in the present invention. For example, acid salts of compounds containing amines or other basic groups can be prepared using suitable organic or inorganic salts. The compound is obtained by reacting an organic acid with the compound to provide a pharma- ceutically acceptable anionic salt form. Examples of anionic salts include acetate, benzenesulfonate, benzoate, diethyl ether, and the like. Carbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride Salt, citrate, dihydrochloride, edetate, edisylate, estrus, escillate Late, fumarate, gluceptate, gluconate, glutamate, glycolylate Glycollylarsanilate, Hexylresorcinate ( hexylresorcinate), hydrobromide, hydrochloride, hydroxynaphthoic acid Salt, iodide, isethionate, lactate, lactobionate, malate, maleate Salt, mandelate, mesylate, methyl sulfate, mucate, naphthyl Salts, nitrates, pamoates, pantothenates, phosphates / diphosphates, polygalacturonates , salicylates, stearates, basic acetates, succinates, sulfates, tannates, Tartrate, tetrahydrofuran, tosylate, triethiodide, and trifluoroacetate salts include.

[0093] Salts of compounds containing an acidic functional group can be prepared by reacting with a suitable base. Such pharma- ceutically acceptable salts can be prepared by adding a pharma- ceutically acceptable cation. This can be made with bases such as alkali metal salts (especially sodium and and potassium), alkaline earth metal salts (especially calcium and magnesium), aluminum and ammonium salts as well as trimethylamine, triethylamine, mol Pholine, pyridine, piperidine, picoline, dicyclohexylamine, N,N'-dibenzylamine Diethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl) Amines, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, Dehydroabietylamine, N,N'-bisdehydroabietylamine, glucamine, N - Physiologically acceptable compounds such as methylglucamine, collidine, quinine, quinoline, etc. The glycerol derivatives are made from organic bases and basic amino acids such as lysine and arginine. Contains salt.

[0094] Pharmaceutical Compositions The present invention relates to cationic polypeptides, PEG-carboxylic acids, and polypeptides having 10 or more carbon atoms. an anionic excipient selected from fatty acids, anionic phospholipids, and combinations thereof; The present invention further relates to a pharmaceutical composition comprising an ionic complex comprising the ionic complex and a pharma- ceutically acceptable carrier. In one embodiment, the pharmaceutical composition may further comprise an additional excipient, such as a carbo For example, the cellulose acetate copolymers described herein may further include hydroxymethylcellulose (CMC). All of the anionic phospholipids used were combined with mPEG-2000-DSPE and CMC. For example, it can be combined with carboxymethylcellulose (CMC).

[0095] The ratio of the concentrations of the cationic peptide and the anionic excipient is based on the charge of the anionic excipient. It is determined with respect to the molar ratio of cationic charges in the polypeptide. For example, For one positive charge, the amount of anionic excipient ranges from 1:1 to 1:10. The ratio of anionic excipients to additional positive charges in the polypeptide may be may be adjusted accordingly. By varying the amount of anionic excipient within this ratio, Thus, the in vivo release characteristics of the polypeptide may be modulated. Typically, a ratio of 100 to 150 typically results in a slower release of the polypeptide from the site of administration than a ratio of 100 to 150, which ... This can result in a composition that provides a fast release.

[0096] The term "pharmaceutical acceptable carrier" in the pharmaceutical composition of the present invention means a biocompatible The polar nature of the liquid means that it remains complex in its ionic form. The biocompatible polar liquid is PEG (polyethylene glycol, e.g., 1 polyethylene glycol having an average molecular weight of 00 to 5000), polyol (e.g. Propylene glycol (PG), tripropylene glycol, glycerol, ethanol benzyl alcohol, DMSO, NMP, DMF, water, pH buffer solutions, and mixtures thereof Further dilutions and further mixtures as described below. It will be appreciated that excipients can be included in the pharmaceutical composition.

[0097] In certain embodiments, the pharmaceutical compositions of the present invention form a drug depot when injected into a subject. In some embodiments, the drug depot comprises a drug that remains in the body for a period of time after injection into a subject. In one embodiment, at least one portion of the pharmaceutical composition is When injected into a subject, it precipitates to form a drug depot and acts pharmacologically over a period of time. In some embodiments, the compositions of the present invention release the active compound in vivo. and a drug depot for sustained, steady-state release of therapeutically effective levels of the polypeptide. This allows for high concentrations of the polypeptide of formula (I) that are suitable for producing An exemplary range of concentration of the polypeptide of structural formula (I) in is about 0.0001 mg / mL. ~ about 100 mg / mL.

[0098] The pharmaceutical compositions of the present invention may contain further excipients (herein referred to as co-excipients). Suitable examples of further excipients include those described herein. pH stable buffers, preservatives, surfactants, stabilizers, antioxidants as defined in the specification , isotonicity agents, and ionic and non-ionic polymers.

[0099] Such auxiliary excipients are ionic complexes or further anionic excipients of the hydrophobic class. For example, added to aid in the formation of homogeneous suspensions and dispersions of lipids or fatty acids. The classes of auxiliary excipients are lecithin, soybean oil, castor oil, migliore migliol, polyethylene glycol (M ranging from 200 to 5,000) Dispersants and additives such as cellulose acetate, methylcellulose, and carboxymethylcellulose. Contains emulsifiers.

[0100] As used herein, the term "surfactant" refers to a surfactant that is a surfactant of the liquid in which it is dissolved. It refers to a surface active agent or substance that tends to reduce surface tension. Suitable surfactants are poly Resorbate, Poloxamer, Triton, Sodium Dodecyl Sulfate, Sodium Lauryl Sulfate For example, surfactants include polyoxyethylene (20 ) sorbitan monolaurate (e.g., Tween® from Sigma-Aldrich) Polyoxyethylene (20) sorbitan monopalmitate (Tween ( Polyoxyethylene (20) sorbitan monooleate (Tween (registered trademark) 80), Poloxamer 188, Polyoxyethylene-Polyoxypropylene Block copolymers (e.g., Pluronic® from Sigma-Aldrich) F-68), polyethylene glycol 660-12-hydroxystearate (So luto 1 (R) HS 15, BASF), Cocoamidopropyl Betaine, Linoleic Acid Irbetaine, myristyl betaine, cetyl betaine, polyoxyethylene castor oil (C Remophor® (now Kolliphor BASF), and lecithin Includes.

[0101] As used herein, the term "tonicity agent" refers to an agent that is used to adjust the tonicity of a formulation. Tonicity generally refers to the osmotic pressure of an aqueous solution compared to that of human serum. The formulation can be hypotonic, isotonic, or hypertonic. Preferably, it is isotonic. Isotonic formulations can be prepared by converting the formulation into a liquid or solid form, e.g., a lyophilized form. It is a liquid that is reduced from a normal state, and is compared to physiological saline and blood serum. The tonicity of the solution is the same as that of some other solution that is injected. This can help reduce pain and irritation in a subject. Dextrose, glycerin, hydroxyethyl starch, lactose, mannitol (e.g. D-mannitol), raffinose, sorbitol, sucrose, trehalose Contains sodium chloride, calcium chloride, magnesium chloride, and potassium chloride .

[0102] As used herein, the term "buffer" refers to an agent that stabilizes the pH of a pharmaceutical composition. Suitable buffers are well known in the art and are described in the literature. Examples of suitable buffers include histidine buffer, citrate buffer, Phar, succinate buffer, acetate buffer, and phosphate buffer or a mixture The most preferred buffers include citrate, L-histidine, or L-histidine. Other preferred buffers include a mixture of acetate and L-histidine hydrochloride. Regardless of the buffer used, the pH should be adjusted according to the methods known in the art. Acids or bases that are used in the manufacture of the product, such as hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, and citric acid, sodium hydroxide, The acid value can be adjusted with sodium hydroxide, potassium hydroxide, or potassium hydroxide.

[0103] Suitable ionic polymers for use as further excipients are ionic carboxymethyl Cellulose (CMC), hyaluronic acid, poly(glutamic acid), poly(aspartic acid) , poly(glutamic acid-co-glycine), poly(aspartic acid-co-glycine), poly (glutamic acid-co-alanine), poly(aspartic acid-co-alanine), starch lycolate, polygalacturonic acid, poly(acrylic acid), carrageenan, and alginate Contains acid.

[0104] In one embodiment, the CMC has an average molecular weight range of about 5,000 to about 700,000. In some embodiments, methyl cellulose and carboxymethyl cellulose are , enhances a sustained release drug depot of an active compound, over a period of time when injected into a subject. It may also help to adjust the viscosity of the ionic complex to release the active compound.

[0105] Suitable non-ionic polymers for use as further excipients are those having a molecular weight of about 100 to 100,000. For example, about 100 to about 5000, about 100 to about 10,000 The polyethylene glycols include those having MW ranging from about 100 to about 60,000. Other weight ranges include from about 200 to about 60,000. A suitable example of a neutral polymer is PE Polyethylene glycol polymers include PE-3350 and PEG-3400. Both G and mPEG are either monomethoxy or dimethoxy. PEG can be structurally represented as follows: HO-(CH2CH2O) n -CH2CH2-OH The value of "n" varies with molecular weight. For example, PEG-3500 acid has a molecular weight of 3,500 The molecular weight of

[0106] mPEG can have one of the following two structures: Monomethoxy mPEG: CH3O-(CH2CH2O) n -CH2CH2-OH. One end is capped as a methoxy group and the other end is capped with a hydroxyl (OH) group. Yes.

[0107] Dimethoxy mPEG: CH3O-(CH2CH2O) n -CH2CH2-OCH3.

[0108] Both ends are capped with methoxy groups.

[0109] The diluent may be used to further dissolve or suspend the ionic complex of the present invention. The diluent may be added to the pharmaceutical composition. Contains biologically compatible materials. Low viscosity non-aqueous injectable liquids include castor oil, vegetable oil, mineral oil, etc. Oil, squalene, monoglycerides, diglycerides, triglycerides, or glycerides In some embodiments, the diluent comprises Miglyol® 812 (Sasol GmbH, Germany), Labrafac® WL13 49 Caprylic acid triglyceride from Gattefosse company, France (Lipoid company, Germany) or Lipoid MCT (Lipoid company, Germany) ) and others of the same kind.

[0110] As used herein, a "drug depot" refers to a portion of a body that is in contact with the body after injection of a pharmaceutical composition of the present invention. The composition refers to a precipitate that may form during administration of the active compound when injected into a subject. A drug depot can be formed, releasing the active compound over a period of time. In the present invention, at least one portion of the composition, when injected into a subject, precipitates and dissolves in water over a period of time. The pharmacologically active compound is released over a period of time.

[0111] "Preservatives" are used to reduce bacterial activity or undesirable chemical changes in a formulation. A compound that can be added to the formulation. Examples of preservatives are benzyl alcohol, ethanol, etc. Alcohol, Methanol, Isopropanol, Butylparaben, Ethylparaben, Methylparaben , propylparaben, catechol, 2-chlorophenol, m-cresol, phenol Resorcinol, Xylitol, 2,6-Dimethylcyclohexanol, 2-Methyl -2,4-Pentanediol, Polyvinylpyrrolidone, Benzethonium chloride, Merciole Thimerosal, Benzoic acid, Benzalkonium chloride, Chlorobutanol, Sodium benzoate These include thorium, sodium propionate, and cetylpyridinium chloride.

[0112] Treatment methods The present invention relates to an ionic complex and a cationic polypeptide of the ionic complex, and the pharmacological activity thereof. Ionic Complexes for Treating Subjects Suffering from Diseases or Disorders Responsive to Sex - Patent application The present invention also relates to the use of a pharmaceutical composition comprising the compound of formula (I) or (II) in the treatment of a disorder to be treated. The method is responsive to modulation of MC4R in a subject in need of treatment. The MC4R modulators, such as those described herein, are cationic polypeptides. In certain embodiments, the method comprises administering to the subject an effective amount of an ionic complex comprising ... Impaired response to MC4R regulation is associated with type 1 diabetes, type 2 diabetes, obesity, and insulin resistance. , metabolic syndrome, male erectile dysfunction, female sexual dysfunction, non-alcoholic fatty liver disease , nonalcoholic steatohepatitis, alcoholism, substance abuse disorders including eating disorders, cachexia, inflammation, and anxiety.

[0113] In certain embodiments, ionic complexes and pharmaceutical compositions comprising the cationic polypeptide of formula I The melanocortin receptor (MC4R) and melanocortin receptor (MC1R) are The present invention has higher selectivity and potency for the mitochondrial cell-3 receptor (MC3R). The onotropic complexes and pharmaceutical compositions have the following properties: increased blood pressure effects, increased heart rate, increased response to sexual stimulation Reduces unwanted effects such as increased skin pigmentation and or can be eliminated.

[0114] As used herein, the phrase "a compound responsive to modulation of the melanocortin-4 receptor" refers to a compound that is "Disorder of MC4R" refers to any disorder that can be treated by activating (stimulating) or inhibiting MC4R. Examples of such disorders are described in more detail below.

[0115] As used herein, the term "modulator" refers to a compound that interacts with a target receptor and A compound that affects the biological function of a receptor. Examples of modulators are full agonists, partial agonists, and Includes partial agonists, neutral antagonists, and inverse agonists.

[0116] As used herein, the term "agonist" refers to an agonist that inhibits the activity of its target, here MC When interacting with (e.g., binding to) 4R, the signal transduction of MC4R beyond its basal level is Agonists refer to any chemical compound, naturally occurring or synthetic, that enhances signal transmission activity. A stimulant is a superagonist (i.e., it acts more potently than the endogenous agonist at the target receptor). It is possible to produce a greater maximal response and thus have a compound with efficacy of 100% or more. Compounds that induce a maximal response after receptor occupancy and activation, and full agonists (i.e., Agonists are compounds that are able to activate the receptor but not inhibit it. The MC4R receptor may be a compound that is unable to induce a maximal response in the vasopressin receptor system. Exemplary agonists are described in detail below.

[0117] As used herein, the term "antagonist" refers to an antagonist that inhibits the activity of its target, Upon interacting with (e.g., binding to) MC4R, Agonist refers to any chemical compound that blocks the signaling activity of a compound.

[0118] As used herein, the term "inverse agonist" refers to an antagonist that inhibits the activity of its target, In this case, when interacting with (e.g., binding to) MC4R, the The term refers to any chemical compound that reduces the basal level of signaling activity of a

[0119] As used herein, an "effective amount" refers to an amount that is therapeutically effective to treat a target disorder. or prophylactically sufficient, as an ionic complex or a pharmaceutical composition comprising the ionic complex. An example of an effective amount is typically about 0.0001 ml g / kg body weight to about 500 mg / kg body weight. An example range is about 0.001 mg / kg to about 500 mg / kg of body weight. For example, an effective amount is about 0.005 mg / kg In other embodiments, the range can range from about 0 to about 500 mg / kg. In other embodiments, an effective amount may range from 0.0001 mg / kg to about 5 mg / kg. , about 0.01 mg / kg body weight to 50 mg / kg body weight or 0.01 mg / kg body weight to 20 mg / kg body weight range.

[0120] As used herein, the term "second agent" refers to any of the agents described herein. In combination with the peptides described herein, or exhibit a synergistic effect with the peptides described herein. Any active pharmaceutical ingredient that exhibits a combined effect that is greater than additive (i.e., As used herein, "enhancing a therapeutic effect" refers to Examples of enhanced therapeutic efficacy include those described herein. a reduction in the effective dose of the peptides described herein; This includes extending the therapeutic window. A second agent can be administered, examples of which are described in more detail below.

[0121] The second agent may be administered prior to, simultaneously with, or in combination with the administration of the peptides described herein. Thus, the peptides described herein can be administered either intravenously or at a later time. and the second agent can be administered together in a single formulation or in separate formulations. For example, the compounds described herein can be administered simultaneously or sequentially. The peptide and the second agent described in are administered sequentially in separate compositions. In such cases, the peptides described herein may be administered prior to or after the second therapeutic agent. Moreover, the peptides and second agents described herein can be administered may or may not be administered on a similar dosing schedule. The peptides and second therapeutic agents described herein may be Various half-life periods may be used so that the peptide is administered more frequently than the second therapeutic agent, or vice versa. The present invention may have different time periods and / or may operate on different time scales. The peptides described herein can be followed by a second agent, which can be either Successive applications of the therapeutic agent result in further enhanced therapeutic efficacy. Either the described peptides or the second agent may be administered acutely or chronically. This can be done.

[0122] An effective amount is a compound having MC4R modulator activity or a pharma- ceutically acceptable salt thereof. By simultaneously administering the first amount and the second amount of at least one second agent, In one embodiment, the method or composition of the invention may be implemented as described herein. The peptide and the second agent described above are each administered in an effective amount. (i.e., in amounts that would be therapeutically effective when administered alone). In one embodiment, the peptide and second agent described herein are each It is administered in an amount that alone does not produce a therapeutic effect (a sub-therapeutic dose). The peptides described herein can be administered in an effective amount, and a second agent is administered in a sub-therapeutic dose. The peptides described herein can be administered in sub-therapeutic doses, The two agents are administered in effective amounts. In an exemplary embodiment, The combination of the peptide and the second agent may be any of the peptides or The second agent exhibits an enhanced therapeutic or synergistic effect as compared to the second agent alone.

[0123] The presence of synergy should be determined using appropriate methods for evaluating drug interactions. A suitable method is, for example, the Sigmoid-Emax equation (Holford, NHGand Scheiner, LB, Clin. Pharmacokin et.6:429-453(1981)), Loewe additivity equation (Loewe, S. and Muischnek, H., Arch. Exp. Patho Pharmacol. 114:313-326(1926)), and the half-effect equation Formula (Chou,TCand Talalay,P.,Adv.Enzyme Reg ul.22 27-55(1984)). Each of the equations referred to above is Experimental data were analyzed to generate corresponding graphs to aid in the evaluation of the effects of drug combinations. The corresponding graphs related to the above mentioned equations are Concentration-effect curves, isobologram curves, and combination indexes ( combination index) curve.

[0124] As used herein, the term "subject" refers to a mammal, preferably a human. However, there are also animals that require veterinary care, such as companion animals (e.g. cats, dogs, cats and the like), livestock (e.g. cows, sheep, pigs, horses, and other similar species), and laboratory animals (e.g., rats, mice, guinea pigs, and "" can also mean "the term "" (and the like).

[0125] As used herein, "treatment" refers to the alleviation of the progression of clinical symptoms associated with a target disorder. For example, " "Treatment" means achieving, partially or substantially, one or more of the following results: To reduce or completely reduce the Type 2 diabetes, prediabetic conditions, hemoglobin A1C (Hb1Ac) of 6% or more ) blood levels, hyperinsulinemia, hyperlipidemia, insulin insensitivity, glucose intolerance ameliorating or improving clinical symptoms or indicators associated with obesity, such as; Slowing, inhibiting, or preventing the progression of obesity and obesity-related symptoms; or partially or completely prevent the onset or development of obesity or obesity-related symptoms. Slowing, inhibiting, or preventing the progression of obesity. Reducing or preventing can, for example, slow the progression of obesity in a normal weight subject. The term "treating" includes preventing, inhibiting, or inhibiting metabolic syndrome. Risk of coronary artery disease, stroke, and diabetes (e.g., type 2) associated with dromes Partially or completely reducing any one or more of the five listed above Metabolic syndrome-related indicators such as It further includes ameliorating or improving clinical symptoms or signs. "Treatment of metabolic syndrome, including insulin resistance and glucose clearance" ROHM-related parameters as well as heart rate and blood pressure, joint disease, inflammation, sleep apnea Other eating disorders, including binge eating, binge eating and bulimia; adjunctive therapy for weight loss surgery; and progression of cardiovascular disease parameters, including adjunctive weight loss therapy prior to orthopedic surgery. "Prophylactic treatment" includes delaying, inhibiting, or preventing the onset of a target disorder. Treatment to prevent, inhibit, or reduce the presence of a disease prior to the onset of clinical symptoms Point.

[0126] Impaired responsiveness Disorders responsive to modulation of MC4R and more generally, the polypeptides listed above Examples of disorders responsive to the pharmacological action of drugs and specific polypeptides are general inflammation, inflammatory Acute and chronic inflammatory diseases such as enteropathy, encephalitis, sepsis, and septic shock Autoimmune components such as rheumatoid arthritis, gouty arthritis, and multiple sclerosis Obesity, eating disorders, and weight gain such as Prader-Willi syndrome Associated metabolic and medical conditions; anorexia, bulimia, and AIDS in frail elderly Metabolic and medical diseases involving weight loss such as wasting, cachexia, cancer cachexia and wasting Diabetes and diabetological related conditions d) conditions and complications of diabetes such as retinopathy; such as skin cancer and prostate cancer. Neoplastic growths; endometriosis and uterine bleeding in women, sexual dysfunction, erectile dysfunction, and Reproductive or sexual medical conditions such as decreased sexual response in women; organ transplant rejection Treatment or injury to the organism, such as ischemia, and reperfusion injury, spinal cord injury and wound healing Treatments to accelerate wound healing, as well as chemotherapy, radiation therapy, temporary or permanent Diseases or conditions resulting from weight loss caused by immobilization or dialysis; bleeding vascular shock, cardiogenic shock, hypovolemic shock, cardiovascular failure, and cardiac cachexia Cardiovascular diseases or conditions such as; acute respiratory distress syndrome, chronic obstructive pulmonary disease, asthma and lung diseases or conditions such as pulmonary fibrosis; associated with allergies or organ transplant rejection To increase immune tolerance and fight attacks against the immune system, such as those that cause psoriasis, skin Skin pigmentation depletion, acne, Treatment of dermatological diseases and conditions such as keloid formation, and skin cancer; pain perception Regulating and treating neuropathic pain, anxiety, depression, memory, and memory dysfunction, etc. behavioral, central nervous system, or neuronal conditions and disorders; alcohol consumption , alcohol abuse, and / or conditions and diseases related to alcoholism; and renal conditions or diseases, such as treatment of renal cachexia or natriuresis Further examples include thyroxine release, aldosterone synthesis and release, body temperature, blood pressure, heart rate number, vascular tone, cerebral blood flow, blood glucose level, bone metabolism, bone formation or development, ovarian weight, Placental development, Prolactin and FSH secretion, Intrauterine fetal growth, Parturition, Spermatogenesis, Sebum and pheromone secretion, neuroprotection and neurogrowth, as well as motivation, learning, and other The invention also includes normalizing or homeostatic activity in a subject, including regulating behavior. This includes binge eating, bulimia, or other eating disorders.

[0127] In an exemplary embodiment, the disorder responsive to modulation of the MC4R receptor is type 1 diabetes, type 2 diabetes, Diabetes, obesity, insulin resistance, metabolic syndrome, cardiovascular disease, or low High density lipoprotein / high density lipoprotein triglyceride imbalance, non-alcoholic fatty liver disease, and substance abuse disorders.

[0128] In an exemplary embodiment, the disorder responsive to modulation of the MC4R receptor is type 1 diabetes, type 2 diabetes, diabetes, obesity, insulin resistance, or metabolic syndrome.

[0129] obesity As used herein, the term "obesity" refers to a weight loss of about 30 kg / m 2 or more Above, for example, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37kg / m 2 vs. those with a body mass index (BMI) of 10.0 or higher In certain embodiments, the obese subject is Have a BMI within the range defined as "obese" by the Control. Last accessed URL on October 28, 2016: http: / / www.cdc.gov / See obesity / defming.html. For example, some implementations In terms of morphology, BMI >= 30.0 kg / m 2 Adults with obesity are obese.

[0130] Diabetes and Related Disorders In an exemplary embodiment, the subject treated by the methods provided herein is a patient suffering from diabetes. "You are at increased risk of having or developing a diabetes-related disorder." The disorders were diabetes mellitus (type 1 (OMIM 222100) and type 2 (OMIM 12585 3)), insulin resistance, and metabolic syndrome.

[0131] In an exemplary embodiment, the subject to be treated is a patient suffering from diabetes (type 1 or type 2); In an exemplary embodiment, the patient has: insulin resistance, or metabolic syndrome. The disorder is diabetes, e.g., type 2 diabetes. In an exemplary embodiment, the subject is "Definition and diagnosis of diabetes mellitus and inter Mediated hyperglycaemia” at the World Health Organization Organization and International Diabetes Federation Risk of having or developing type 2 diabetes as defined by deration In an exemplary embodiment, a diabetic subject has a fasting Blood glucose level or 2-hour plasma glucose (2 hours after oral administration of 75 mg glucose) >= 2 In an exemplary embodiment, a diabetic or prediabetic subject is Increased levels of glycated hemoglobin, e.g., 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, 6.0, 6.2, 6.4, 6.6.6.8, 7.0, 7.2, 7. In an exemplary embodiment, the diabetic and pre-diabetic The subject of interest has a genetic polymorphism in or near one or more of the genes in Table 1 below. For example, alterations in expression levels, e.g., expression levels in coding sequences and / or and may be identified or further characterized by a polymorphism that results in an increase or decrease in a mutation. It may be possible to do so. [Table 2] TIFF2025004067000020.tif34128

[0132] In an exemplary embodiment, a subject to treatment by the methods provided by the present invention Further genes that can be used to identify or characterize a subject include FTO (OMIM 610966), JAZF1 (OMIM 606246), and HHEX (OMIM 604420).

[0133] In an exemplary embodiment, a subject to treatment by the methods provided by the present invention The subject has type 1 diabetes. In an exemplary embodiment, the subject with type 1 diabetes is Peptide assays, e.g., less than about 1.0 nmol / L, e.g., 1.2, 1.1, 1. Less than 0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4 nmol / L, e.g. 0. Fasting C-peptide levels below 33, 0.25, 0.2, or 0.1 nmol / L In an exemplary embodiment, C-peptide levels are characterized as a function of oral glucose After the challenge (2 hours after oral administration of 75 g glucose) Measured and below 0.54 nmol / L, e.g. 0.50, 0.45, 0.40, 0.3 Increase of less than 5, 0.30, 0.25, 0.20, 0.15, or 0.10 nmol / L Impaired fasting glucose (110-125 mg / dL) or impaired glucose tolerance (75 g 2 hours after oral glucose challenge: 140-199 mg / dL) To identify or further characterize the decline in beta cell function in subjects with diabetes In an exemplary embodiment, a type 1 diabetes patient is treated with islet cell antigens and / or or autoantibodies against insulin, e.g., 65 kDa GAD (OMIM 138 275) and / or autoantibodies directed against the phosphatase-associated IA-2 molecule. Identified or further characterized by the presence of

[0134] Insulin resistance In an exemplary embodiment, the disorder is diagnosed by any means known in the art. may be identified, characterized by a reduced ability of insulin to lower blood glucose levels. In an exemplary embodiment, insulin resistance is characterized by or further identified by the presence of one or more polymorphisms in one or more of the following genes: (e.g., polymorphisms that lead to altered expression levels, e ... (including variants of coding sequences for gene products such as proteins and the like, and variants of coding sequences for gene products such as proteins and the like): ETN, PTPN1, TCF1 (OMIM 142410; see e.g. polymorphism 0011) ), PPP1R3A (OMIM 600917; e.g., polymorphisms 0001, 000 3), PTPN1 (OMIM 176885; see e.g., polymorphism 0001), see for example polymorphism 0006), ENPP1 (OMIM 173335; see for example polymorphism 0006 ), IRS1 (OMIM 147545; see e.g., polymorphism 0002), EPHX2 (OMIM 132811; see e.g., polymorphism 0001), (OMIM 164160, see e.g. polymorphisms 0001 and 0002), Leptin receptor (OMIM 601007, e.g. polymorphisms 0001, 0002, 0004 , and 0005), or insulin receptor (INSR, OMIM 1 47670, see e.g. polymorphisms 0001-0037).

[0135] Metabolic syndrome In an exemplary embodiment, the disorder is metabolic syndrome. As used, the term "metabolic syndrome" refers to a set of conditions that occur together and are associated with coronary artery disease, stroke, and other conditions. American Heart Association (AMEA) refers to a group of conditions that increase the risk of developing type 2 or 3 diabetes. Can Heart Association and National Heart,L According to the U.S. and Blood Institute, the condition is called Metformin Syndrome (also known as syndrome X). Tabooic syndrome is present when a subject has three or more of the following signs: 1) Blood pressure equal to or higher than 130 / 85 mmHg; 2) fasting blood sugar (glucose) equal to or greater than 100 mg / dL; 3) Large waist circumference (the length around the waist): - Males - over 40 inches; -Females - 35 inches and above; 4) Low HDL cholesterol: -Men - less than 40 mg / dL; -Women - Less than 50 mg / dL; 5) Triglycerides equal to or greater than 150 mg / dL.

[0136] Metabolic syndrome is characterized by changes in a subject's blood pressure, blood glucose levels, HDL cholesterol, cholesterol levels, LDL cholesterol levels, total cholesterol levels, and triglycerides This can be diagnosed by testing blood levels.

[0137] In an exemplary embodiment, the subject is diagnosed with central obesity (waist circumference >= 80cm for women). m;>=90cm for Asian men, including Latin American men, and for all other men >=94cm), BMI >30kg / m 2 , elevated triglycerides (>=150 mg / dL or specific treatment for this lipid abnormality), lowering HDL cholesterol (male <40 mg / dL in men and <50 mg / dL in women, or specific treatment for hypertension), elevated blood pressure (sBP>=130mmHg or dBP>=85 mmHg or treatment of previously diagnosed hypertension) or elevated fasting plasma glucose (FPG> = 100 mg / dL or previous type 2 diabetes diagnosis), including any combination thereof. In an exemplary embodiment, the subject to be treated by the methods provided herein is The elephant was featured in The IDF, published in 2006, which is incorporated by reference in its entirety. worldwide consensus definition of the m etabolic syndrome” at the International Diab Metabolic syndrome, as defined by the Etes Federation The subject may have or be at increased risk of having central obesity (as described above). and / or BMI > 30 kg / m 2 ) and elevated triglycerides, Either a decrease in HDL cholesterol, an increase in blood pressure, or an increase in fasting blood glucose In an exemplary embodiment, metabolic syndrome is characterized in that the subject has 27 (see, e.g., OMIM 605552) and / or 17p12 (see, e.g., OMIM 605552) Mutations at loci selected from the following: Characterized or further characterized by the presence

[0138] Disorders caused by MC4R mutations The present invention relates to a method for determining the response of MC4R to α-melanocortin stimulating hormone (α-MSH). The present invention relates to a method for treating a disorder in a subject suffering from attenuation, the method comprising administering to a subject a melanocortin-containing The method includes administering an effective amount of an agonist of the mitochondrial cell 4 receptor (MC4R). In some embodiments, the subject has a MC for α-melanocortin stimulating hormone (α-MSH). Heterozygous carriers of MC4R mutations that result in attenuated 4R responses. MC4R is a cellular signaling molecule that is capable of signaling to MC4R receptors, since the conjugated carriers retain the ability to respond to natural ligands of MC4R. Administration of an agonist corrects MC4R-associated disorders in heterozygous carriers. Treatment is independent of knowledge of the type of MC4R mutation.

[0139] In an exemplary embodiment, the disorder is obesity, e.g., MC4R-associated obesity. In an exemplary embodiment, the disorder is metabolic syndrome.

[0140] The human MC4R gene (hMC4R) has the GenBank accession number CH471077. It is a well-characterized protein encoded by a genomic sequence that corresponds to the

[0141] Mutations in the MC4R receptor are a related cause of severe childhood obesity. The carrier prevalence for MC4R mutations in the obese population is approximately 2.5%. was noted, with the highest prevalence at 6% among severely obese children. Humans with mutations are compared to mice with mutations in the MC4 receptor gene. As described, they show more or less the same phenotype. levels, gonadotropin, thyroid, and sex steroid levels unchanged, and lean body weight , bone mineral density, and linear growth velocity with apparent hyperphagia, hyperinsulinemia, and body fat mass. In contrast to MC4 receptor deletion, hyperphagia and hyperinsulinemia are not observed in humans. There is a tendency for sedation to occur with age in subjects. The phenotype in telozygous carriers is moderate compared to homozygous carriers. The hyperphagia observed in experimental feeding studies was observed in people with leptin deficiency. MC4 receptor dysfunction seen in vitro in assays The severity of the mutation depends on the amount of food ingested in the test meal by subjects carrying that particular mutation. At least 90% of obesity-related symptoms are predictive and correlate with the onset and severity of the obesity phenotype. Various MC4 receptor mutations are associated with obesity, and further mutations in the MC4 receptor , are likely to be found and lead to a similar obesity phenotype.

[0142] Examples of MC4R mutations that cause obesity in humans are included, relevant parts, by reference. Farooqi et al., The Journal of l of Clinical Investigation,July 2000,vo l.106(2), pp.271-279 and Vaisse et al.,The Journal of Clinical Investigation,July 2 000, vol. 106(2), pp. 253-262.

[0143] Further mutations that potentially cause obesity in humans are involved Xiang et al., "Pharmacokinetics and Biochemical Properties," incorporated herein by reference. logical characterization of 30 human mel anocortin-4 receptor polymorphisms with the endogenous proopiomelanocortin-deriv ed agonists,synthetic agonists,and the e endogenous agouti-related protein antagonist ist.”Biochemistry,2010 Jun 8;49(22):4583 As described in -600, R18H, R18L, S36Y, P48S, V50 M, F51L, E61K, I69T, D90N, S94R, G98R, I121T, A1 54D, Y157S, W174C, G181D, F202L, A219V, I226T, G231S, G238D, N240S, C271R, S295P, P299L, E308 K, I317V, L325F, and 750DelGA.

[0144] Further examples of mutations that potentially cause obesity in humans are found at the URL http: / / www.htt Accession number 155541 (MC4 R) (more precisely, under accession numbers 155541.0001-155541.0023) Online Mendelian Inheritance in Man(OMIM ), as listed in the Human Genes and Genetic Diseases Database. Representative examples 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;1 5-BP DEL (delta 88 to 92 codons); and SER127LEU. The relevant portions of the OMIM database are incorporated herein by reference. .

[0145] In an exemplary embodiment, the MC4R mutation results in retention of MC4R signaling activity. Glass.

[0146] Mutations in the genomic sequence encoding MC4R can be made using methods well known to those skilled in the art. For example, genomic sequences can be detected by methods such as those described in, for example, Farooqi et al. et al,The Journal of Clinical Investigat ion,July 2000,vol.106 (2),pp.271-279 and V aisse et al, The Journal of Clinical Inves. stigation,July 2000,vol.106(2),pp.253-26 Cloning is performed using nucleotide primers such as those described in 2. The cloned sequences can be sequenced using commercially available sequencers and and software can be used to analyze.

[0147] The activity of MC4R can be measured by methods well known to those skilled in the art. For example, cells can be transiently transfected with cloned MC4R DNA. and contacting the transfected cells with an agonist of MC4R (e.g., α-MSH). This increases the intracellular level of cAMP, a second messenger of MC4R, for example by increasing the bert et al, Journal of Endocrinology (2010 ) 207, pp. 177-183, measured by the electrochemiluminescence assay The reduction in MC4R signaling may be due to the lack of a given agonist by wild-type MC4R. The intracellular levels of cAMP produced in response to stress were compared with those produced by mutant MC4R. This can be verified by comparing the levels

[0148] MC4R modulators (eg, agonists) also inhibit the normal function of the natural agonist of MC4R. It may also be used to treat patients suffering from other disorders such as depression. Examples of such patients include those with leptin-dependent pathways (Nature Clinical Practice tice Endocrinology and Metabolism,2006;2 ;6;318 and NEng J Med:2007;356;3;237), Pro Opiomelanocortin processing (Nature Genetics, 1998, 15 5;Cell Metabolism,2006;3;135;Annals Acad Med, 2009, 38;1;34), or encoding a prohormone convertase Heterozygous or homozygous for mutations in important genes This includes individuals who are monozygous.

[0149] Mode of Administration The methods described herein are useful for carrying out the methods described herein. Administration of the ionic complex or pharmaceutical composition may be continuous, hourly, four times daily, three times daily, Twice daily, once daily, once every other day, twice weekly, once weekly, once every two weeks, once a month or once every 2 months, once every 3 months, once every 4 months, or once every 5 months or once every 6 months or more, or some other intermittent dosing schedule The ionic complex composition of the present invention can be administered once daily, once weekly, or once every two weeks. once every 4 weeks, once every 2 months, once every 3 months, once every 4 months, Suitable for effective therapeutic dosing over a range of once every 5 months or once every 6 months There are.

[0150] Suitable methods of administration include, but are not limited to, peripheral administration. An example of peripheral administration is , Administration of oral, subcutaneous, intraperitoneal, intramuscular, intravenous, rectal, transdermal, buccal, sublingual, inhalation, pulmonary, intranasal A preferred embodiment uses subcutaneous administration.

[0151] Combination therapy Any peptide described herein may be part of an ionic complex or uncomplexed (e.g. For example, the peptide may be a peptide pre-complexed to one or more other pharma- ceutical active compounds. (the "second agent") to inhibit any of the disorders responsive to modulation of MC4R. Such combined administration can be used for the treatment of any of the following conditions: and one or more second drugs. Such single dosage forms may be tablets, capsules, aerosols, inhalation powders, etc. Alternatively, the combination administration may be in the form of two This can be by administration of two different dosage forms, one of which is a dosage form as described herein. Other dosage forms contain one or more peptides as described in the above, and one or more second agents. In this case, the dosage forms may be the same or different. Although not meant to be limiting, the following combination therapies may be used: Here, we illustrate some combination therapies.

[0152] The peptides described herein (e.g., as part of an ionic complex or uncomplexed) (as) are associated with various weight and eating-related conditions, such as obesity and / or overweight It may be combined with one or more second agents useful in the treatment of disorders. Drug 2 acts on energy consumption, glycolysis, gluconeogenesis, glycogenolysis, lipolysis, lipogenesis, Fat absorption, fat storage, fat excretion, hunger and / or satiety and / or craving mechanisms Anti-obesity drugs that affect rhythm, appetite / motivation, food intake, or gastrointestinal motility Drugs that reduce energy intake may be used in conjunction with behavioral therapy in weight loss programs. These include a variety of pharmacological agents called appetite suppressants that are used as adjuncts to appetite suppressants.

[0153] Generally, the total dosage of the obesity control agent or pharmaceutical agent will be within the range of 1 to 4 mg / kg of the dose described herein. When used in combination with one or more peptides, administer in a single or 2-4 divided doses. 0.1 to 3,000 mg / day, preferably about 1 to 1,000 mg / day, more preferably about This can range from 1 to 200 mg per day. However, the exact dose should be determined by your doctor. The dosage is determined by the potency of the compound administered, the age, weight, condition, and response of the patient, etc. It depends on factors such as:

[0154] One or more peptides described herein (part of an ionic complex or uncomplexed) The compound (as a diabetic retinopathy) may be combined with one or more second agents useful in the treatment of diabetes. This can be done.

[0155] One or more of the peptides described herein may also or alternatively be insulin resistance; impaired glucose tolerance; type 2 diabetes; metabolic syndrome; dyslipidemia (hyperlipidemia) hypertension; heart disease (e.g. coronary heart disease, myocardial infarction); cardiovascular disorders Nonalcoholic fatty liver disease (including nonalcoholic steatohepatitis); Joint disorders (secondary (including osteoarthritis); gastroesophageal reflux; sleep apnea; atherosclerosis; stroke; major and microvascular disease; steatosis (e.g., in the liver); gallstones; and gallbladder disorders. For the treatment of obesity and / or overweight related diseases, disorders, and / or conditions, such as It may be further usefully combined with one or more second agents.

[0156] Second Agent The one or more second agents can be, for example: Insulin and insulin analogues; Sulfonylureas (e.g., glipizide) and meglitinides (e.g., repaglinide) and nateglinide) (sometimes called "short-acting secretory insulin secretagogues, including insulin secretagogues (also called "insulin secretagogues"); Agents that improve incretin action: Incretins, incretin mimetics, incretins Agents that improve cretin function, e.g., GLP-1, GIP; GLP-1 agonists (e.g. exenatide and liraglutide (VICTOZA)), DPP-4 inhibitors ( (e.g., vildagliptin, saxagliptin, and sitagliptin) Permethrins, such as thiazolidinediones (e.g., pioglitazone and rosiglitazone), Insulin sensitization with oxisome proliferator-activated receptor gamma (PPARγ) agonists Agents having any combination of PPAR alpha, gamma, and delta activity. Substances that: Agents that regulate hepatic glucose balance, such as biguanides (e.g., methotrexate, Lumin), fructose 1,6-bisphosphatase inhibitor, glycogen phosphorylator glycogen synthase kinase inhibitors, glycogen synthase kinase inhibitors, and glucokinase activators; Alpha-glucosidase inhibitors (e.g., miglitol and acarbose) Agents designed to reduce / delay glucose absorption from the unguinea pig intestine; Antibody that antagonizes the action of glucagon, such as amylin analogues (e.g., pramlintide), Agents that nicotine or reduce its secretion; Sodium-dependent glucose transporter type 2 (SGLT-2) inhibitors (e.g., dapagliflozin) Agents that prevent glucose reabsorption by the kidney, such as rifapril; Aldose reductase inhibitors (e.g., epalrestat and ranirestat), etc. Agents designed to treat complications of long-term hyperglycemia such as; Actions to treat complications related to micro-angiopathy substance for use; HMG-CoA reductase inhibitors (statins, e.g., rosuvastatin) and other Anti-dyslipidemic agents such as cholesterol lowering agents; PPARα agonists (fibrates, e.g. gemfibrozil and fenofibrate ; Bile acid sequestrants (e.g., cholestyramine); Cholesterol absorption inhibitors (e.g., plant sterols (i.e., phytosterols), synthesis inhibitors); Cholesteryl ester transfer protein (CETP) inhibitors; inhibitors of the ileal bile acid transport system (IBAT inhibitors); bile acid binding resin; Nicotinic acid (niacin) and its analogues; Antioxidants such as probucol; Omega-3 fatty acids; Beta blockers (e.g., atenolol), alpha blockers (e.g., doxazosin), and and adrenergic receptor antagonists such as mixed alpha / beta blockers (e.g., labetalol). Antihypertensive drugs, including antihypertensive agents; Adrenergic receptor agonists, including alpha-2 agonists (e.g., clonidine); Angiotensin-converting enzyme (ACE) inhibitors (e.g. lisinopril), dihydropyridin amines (e.g., nifedipine), phenylalkylamines (e.g., verapamil), and Calcium channel blockers such as benzothiazepines (e.g., diltiazem); Angiotensin II receptor antagonists (e.g., candesartan); aldosterone receptor antagonists (e.g., eplerenone); Centrally acting adrenergic agonists, such as central alpha agonists (e.g., clonidine) agonists; as well as diuretics (e.g. furosemide); Hemostatic modifiers, including antithrombotic agents such as activators of fibrinolysis; Thrombin antagonists; Factor VIIa inhibitors; vitamin K antagonists (e.g. warfarin), heparin and its low molecular weight analogues, factor Xa inhibitors, and direct thrombin inhibitors ct thrombin inhibitors (e.g. argatroban) anticoagulants; cyclooxygenase inhibitors (e.g. aspirin), adenosine diphosphate (ADP) receptor inhibitors (e.g., clopidogrel), phosphodiesterase inhibitors (e.g., cilostazol), glycoprotein IIB / IIA inhibitors (e.g. tirofiban) and antiplatelet agents such as adenosine reuptake inhibitors (e.g., dipyridamole). medicine; Noradrenergic agents (e.g., phentermine) and serotonergic agents (e.g., Appetite suppressants (e.g., sibutramine), including ephedrine, pancreatic lipase inhibitors agents (e.g., orlistat), microsomal transfer protein (MTP) modifiers, DGAT inhibitors and cannabinoids anti-obesity agents such as CB1 receptor antagonists (e.g., rimonabant); Such as orexin receptor modulators and melanin-concentrating hormone (MCH) modulators. Feeding behavior modifiers; Neuropeptide Y (NPY) / NPY receptor modulators; Pyruvate dehydrogenase kinase (PDK) modulators; Serotonin receptor modulators; leptin / leptin receptor modulators; Ghrelin / ghrelin receptor modulators; Agents that enhance beta cell function; Agents that stimulate energy expenditure (e.g. beta-adrenergic stimulants, UCP-1 agonists, brown fat modulators and stimulators); agents that induce 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 Selective serotonin reuptake inhibitors (SSR1) (e.g. fluoxetine), noradrenaline Renaline reuptake inhibitors (NARIs), noradrenaline serotonin reuptake inhibitors (SNRI), triple monoamine reuptake blockers reuptake blockers (e.g., tesofensine), and monoamine agonists. Amino acid oxidase inhibitors (MAOIs) (e.g., toloxatone and amifluramine) or a pharma- ceutically acceptable salt thereof.

[0157] In an exemplary embodiment, an MC4R agonist (e.g., part of an ionic complex or a non-ionic complex) is The complex (MC4R agonist) and the second agent were administered in a very low calorie diet (VLCD). ) or a low-calorie diet (LCD) administered simultaneously, sequentially, or separately.

[0158] Method for preparing ionic complexes The present invention further relates to methods for preparing the ionic complexes and pharmaceutical compositions of the present invention. The method comprises the steps of: a cationic polypeptide and a PEG-carboxylic acid, The anionic excipient is selected from a fatty acid having a The method includes the step of: reacting a cationic polypeptide and a cation under conditions for forming an ionic complex. and an anionic excipient, and preparing a pharmaceutical composition comprising the ionic complex. A step of manufacturing.

[0159] For example, PEG-carboxylic acids in aqueous media (e.g., water), an anionic excipient selected from a fatty acid, a phospholipid, or a combination thereof, and additional excipients by autoclaving the mixture under suitable conditions. The mixture can be uniformly dispersed. Suitable conditions are, for example, about 3 minutes. An example of suitable conditions may include a temperature of 121° C. to 134° C. for about 1 hour. The use of an autoclave also involves sterilizing the mixture for a period of 5 minutes and a temperature of approximately 121°C. The sterile aqueous solution of the cationic peptide is then mixed with the sterile homogenous ionic complex of the present invention. Depending on the nature of the anionic excipient, it may be added to the mixture to provide a A clear aqueous solution or a homogeneous suspension of the ionic complex can be obtained. PEG carboxylates and mPEs used in the examples described in the specification G2000-DSPE typically results in a clear aqueous solution containing ionic complexes, whereas The use of DPPA lipids typically results in a homogenous suspension containing ionic complexes.

[0160] Alternatively, the mixture of ionic complexes may be dissolved in water with the cationic polypeptide. The agent is dissolved and the resulting mixture is filtered through a 0.2 micron filter. It can be prepared by sterile filtration at 37°C.

[0161] Cationic polypeptides and PEG-carboxylic acids, lipids with 10 or more carbon atoms Ionic complexes comprising an anionic excipient selected from acids, phospholipids, and combinations thereof 1. A method for producing a body, comprising: a) preparing a mixture of an anionic excipient and an aqueous excipient diluent; b) autoclaving the mixture under conditions sufficient to sterilize the excipients; c) mixing a sterile peptide solution containing the cationic polypeptide and an aqueous peptide diluent with an excipient; The method comprises the step of adding to the mixture.

[0162] In one embodiment, the excipient mixture is a suspension. In another embodiment, the excipient mixture is , a solution.

[0163] In another embodiment, the present invention provides a method for the synthesis of a cationic polypeptide and a PEG-carboxylic acid, 10 an anion selected from fatty acids having 1 or more carbon atoms, phospholipids, and combinations thereof; 1. A method for making an ionic complex comprising an ionic excipient, the method comprising: a) preparing a solution of anionic excipient and aqueous excipient diluent; b) filtering the solution of step a through a 0.2 micron filter; c) step 20 of a sterile peptide solution comprising a cationic polypeptide and an aqueous peptide diluent; b) adding to the excipient solution.

[0164] In another embodiment, the present invention provides a method for the synthesis of a cationic polypeptide and a PEG-carboxylic acid, 10 an anion selected from fatty acids having 1 or more carbon atoms, phospholipids, and combinations thereof; 1. A method for making an ionic complex comprising an ionic excipient, the method comprising: a) Prepare an aqueous solution containing an anionic excipient, an aqueous excipient diluent, and a cationic polypeptide. A step of manufacturing b) Sterilize the resulting aqueous solution by filtering through a 0.2 micron filter. The present invention relates to a method comprising the steps of:

[0165] Suitable examples of excipients and cationic polypeptide excipients include polyols (e.g., propylene glycol, propylene glycol ether, propylene glycol terpolymers, propylene glycol ether ... Polyethylene glycol, tripropylene glycol, glycerol, ethanol, benzyl alcohol Coal, DMSO, NMP, DMF, water, pH stable buffer solutions, and mixtures thereof) .

[0166] [Example] Peptide synthesis Cationic polypeptides suitable for use in the present invention can be prepared by conventional solid phase peptide synthesis. For example, peptide 1 and other MC4R analytes described herein were prepared using the same method. The log is disclosed in U.S. Patent No. 8,349,797, the entire contents of which are hereby incorporated by reference. The peptide chain can be prepared according to the method described in the specification of the same application. The solid support resin is coupled to a suitable solid support resin known to be suitable for the synthesis of amides. Starting from the C-terminal amino acid derivative, the C-terminal amino acid was elongated in a stepwise manner. For the synthesis of doped functionalized peptides, Rink amide MBHA resin is the solid support. For the synthesis of peptides with a C-terminal free carboxyl function, 2-Chlorotrityl chloride resin that forms ester bonds with moc-amino acids, Wang Resins such as cellulose acetate or Merrifield resins may be utilized. These ester-linked Fmoc-amino acid resin types are available from a variety of sources. , are generally used where appropriate.

[0167] Synthesis of disulfide-cyclized peptides Linear derivatives of disulfide cyclic peptide amides were synthesized using a solid-phase peptide synthesizer with Fm The Fmoc-Rink amide resin was placed in the reaction vessel and assembled using . It was then filled with NMP and diluted with 20% piperidine in NMP for 15 minutes. The resin was treated and then washed three times with NMP. Tested (Kaiser, E., Colescot, R.L., Bossinge, C. D. & Cook,PIAnal.Biochem.,1990,34:595- 598). It was resuspended in NMP and the first C-terminal Fmoc-amino acid required was added. The derivative and HOBt were mixed. The coupling reaction was carried out using HBTU reagent and DIPEA. After mixing for 2-3 hours, the coupling was complete with 10% CO from the reaction mixture. This was confirmed by a negative Kaiser test on a small aliquot of the removed resin. The resin was washed three times with NMP. The Fmoc group was then cleaved by cleavage as previously described. and the entire cycle is completed as described to afford the second C-terminal Fmoc-amino acid derivative. The same cycle of reaction was repeated over time with each incoming amino acid. The chloranil color test (Vojkovsky, T. Pept. Res., 1999) 95,8:236-237) by Fmoc deprotection of proline residues in peptide sequences. For a positive test, in addition, the completion of the coupling of the amino acid to proline (negative crosslinking) The Ranil test was used instead of the Kaiser test to test the N-terminal acetylated In the case of peptides with pyridine groups, the Fmoc-deprotected peptide resin was reacted with acetic anhydride and pyridine. The resins after testing negative for the Kaiser test were treated with NMDA for 10 min. The Fmoc amino acid derivatives were washed with dichloromethane and dried in vacuum. The following trifunctional amino acid derivatives were used in the synthesis of these peptides: The following were used: Fmoc-Cys(Trt)-OH, Fmoc-Trp(Boc)-OH, Fmo c-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, Fmo c-His(1-Me)-OH, Fmoc-His(3-Me)-OH, and Fmoc -Glu(OBuTo)-OH.

[0168] To cleave the peptide from the resin and deprotect the side chain functional groups, the peptide resin was diluted with 2% The solution was taken up in TIS / 5% water / 5% (w / v) DTT / 88% TFA. The aqueous solution was stirred for 3.5 h. The mixture was mixed for 10 min and then filtered. The filtrate was mixed with cold anhydrous ether. The precipitate was centrifuged. The solvent was decanted and the peptide pellet was resuspended in fresh ether. The ether workup was repeated two more times. The peptide was dried in vacuum. The peptide product was diluted to a concentration of 2 mg / mL in 5% acetic acid and purified using 0.5 M iodine / methanol. The ethanol was added dropwise with vigorous stirring until a stable pale yellow color of the aqueous solution was achieved. The aqueous solution was stirred for an additional 10 minutes. The excess iodine was then removed by stirring until the mixture became colorless. The reaction was quenched by adding 1M sodium thiosulfate under mixing at RT. The solution was lyophilized and the crude powder was purified by preparative HPLC using a reversed phase C-18 column. The purified product fractions were pooled and lyophilized. The peptides were purified by electrospray spectrometry. The compounds were analyzed by mass spectrometry using single ionization techniques and identified and mass corrected.

[0169] Synthesis of lactam cyclized peptides Cyclic lactam peptides were also synthesized by standard solid phase peptide synthesis methods. For peptides with terminal Dpr, Fmoc-Dpr(Mtt)-BHA resin was used as the immobilized The resulting mixture was transferred to a 2-phase peptide synthesizer reactor. The Fmoc group was then added as described above. and the next Fmoc-protected amine, e.g., Fmoc-Trp(Boc)-OH, is The acid was coupled to the resin by standard coupling procedures. The Fmoc protecting group was removed. The remaining amino acids are then subjected to coupling and deprotection procedures until the amino acid sequence is completed. By repeating the process, each was added in the correct order. The ring-Fmoc-Glu (OPip) was then used. The fully assembled peptide was then The disulfide series was synthesized by N-terminally cleaving the peptides using the method described previously for disulfide series of peptides. The orthogonal protected side chains were then removed. For example, 2-phenyl Glu as its isopropyl (OPip) ester or as its 4-methyltrityl (Mtt) ester The peptide resin bearing the orthogonally protected side chain of Dpr was prepared using 1% TFA in dichloromethane. The deprotected peptide resin was suspended in NMP and cleaved with HBTU D After cyclization (negative Kaiser test), the peptide resin was treated with DC The cyclic peptide was purified by washing with water and 1,2-ethanedithiol (E DT) to remove any remaining protecting groups using trifluoroacetic acid (TFA). The product was collected by precipitation upon addition of cold anhydrous ether. The final purification was performed by reversed-phase HPLC using a reversed-phase C-18 column. The purified peptides were collected by lyophilization and electrospray spectrometry. was used and analyzed for its mass by mass spectrometry. [Table 3] TIFF2025004067000022.tif214140TIFF2025004067000023.tif216140TIFF2025004067000024.tif88140

[0170] Certain cationic polypeptides in Table 1 have activity in the assays described below. The data are provided in Table 2.

[0171] Peptide Testing: Radioligand binding assay: The binding constant (K d ) or inhibitory concentration (IC 50 Receptor binding assays for determining This may be done by any means known in the art.

[0172] As an example, cell membrane preparations for binding assays have been prepared using hMC receptor subtypes 1, 3, and 4. , or prepared from CHO-K1 cells transfected to stably express 5 do.[ 125 I](Tyr 2 )-(Nle 4 -D-Phe 7 )-alpha-MSH([ 125 Competitive inhibition of [I]-NDP-α-MSH binding was measured using polypropylene 96-well plates. Briefly, cell membranes (1-10 μg / ml) prepared as described above were used. The cells were incubated with increasing concentrations of test compounds and and 0.1-0.3 nM 125 I]-NDP-α-MSH and 0.2% BSA, 5 mM M MgCl2, 1 mM CaC l 2, and containing 0.1 mg / mL bacitracin Incubate in 50 mM Tris-HCl, pH 7.4. 12 5 I]-NDP-α-MSH ligand was prepared by mixing 0.1% (w / v) polyethyleneimine (PE I) Presoaked GF / C glass fiber filter plate (Unifilter®) The free [ 125 I]-ND The filter is then cooled to 50°C at a pH of 7.4 and a temperature of approximately 0–4°C. The plates are washed three times with 1 mM Tris-HCl and then assayed for radioactivity. The combined data is analyzed by computer-assisted nonlinear regression analysis.

[0173] Cyclic AMP stimulation assay: Functional assays that determine the agonist or antagonist status of the cyclic peptides of the present invention Assays may be performed by any means known in the art.

[0174] Electrochemiluminescence (ECL) assay Stimulation of intracellular cyclic AMP (cAMP) levels by peptides was observed by electrochemiluminescence (ECL) assay. (ECL) assay (Meso Scale Discovery, Gaithersb urg, MD, USA; hereafter referred to as "MSD") in a dose-dependent manner. Briefly, CHO-K1 cells stably expressing hMC receptor subtypes were cultured in vitro and cultured in vitro. Suspend in MPI 1640® Assay Buffer (RMPI 1640 Buffer) The buffer contains 0.5 mM IBMX and 0.2% protein cocktail (MS D-blockers A)). Transgenic mice stably expressing hMC receptor subtypes 1, 3, 4, or 5 were Approximately 7,000 transgenic CHO-K1 cells were cultured per well on an integrated carbon electrode. A 384-well multi-array plate ( Increasing concentrations of test compound are added and the cells are incubated at 37° C. for approximately 40 Incubate for 1 min. Add 0.2% protein cocktail and 2.5 nM TAG (Trademark) ) Cell lysis buffer (pH 7.3, Mg) containing ruthenium-labeled cAMP (MSD) HEPES-buffered saline solution with Cl2 and TritonX-100® and incubate the cells at room temperature for approximately 90 minutes. At the end of the period, add ECL co-reactants in lead buffer (pH 7.8). Add Tris buffer solution (containing ribosomal phosphate buffer and Triton X-100) to lyse the cells. The cAMP levels in the lysates were measured using a Sector Imager 6000 reader (registered trademark). The data were determined directly by ECL detection using the MSDS (MSD) system. Analyzed using assisted nonlinear regression analysis (XL fit; IDBS) and reported as EC50 values. The EC50 is 50% of the maximum reaction response, e.g., in the assay described above. Agonist compounds required to obtain 50% of the maximal level of cAMP as determined using The concentration of is shown.

[0175] cAMP Measurement Assay: Human MC4-R transfected cells were grown to confluence in 96-well plates. Grow cells to 0.2 mL (approximately 250,000 cells plated per well). mM isobutylmethylxanthine (IBMX) and graded concentrations of peptide or surrogate Treat in triplicate sets with peptides in the presence of 20 nM NDP-MSH. Similarly, cells treated with 20 nM NDP-MSH alone were incubated in 200 μL of The volume was used as a positive control. A far blank was also included. After 1 hour incubation at 37°C, the cells were incubated for 5 Lyse the cells by adding 250 μL of cell lysis buffer. Total cAMP accumulated in the reaction medium was determined according to the procedure specified by the kit supplier. A commercially available low pH cAMP assay kit (Amersham Biosciences) was used. Quantification was performed using alpha- Peptides that show the same or higher cAMP accumulation as MSH are considered agonists. The data for the agonists are plotted to determine the EC50 value. The negative control (in the absence of alpha-MSH) Peptides that show accumulation in the same range as the blank buffer (containing 100% ethanol) are ineffective at the concentrations tested. The peptides that showed the attenuation of the product were also present in the assay when alpha-MSH and cAMP If there is inhibition of hMC-1, it is considered an antagonist. This can be performed with hMC-R, hMC-3R, and hMC-5R cells.

[0176] Measurement of cAMP accumulation using the β-galactosidase (β-Gal) reporter system: Enzyme fragment complementation with β-galactosidase (β-Gal) as a functional reporter system (enzyme fragment complementation) (EFC) system A chemiluminescence readout system was used. Assay systems are available (cAMP Hunter GPCR assay system). stem, Discoverx Corp, Fremont, CA). Two complementary parts; EA for the enzyme acceptor and ED for the enzyme donor The assay utilizes the β-Gal enzyme, which is split into two parts: the ED moiety fused to cAMP. The cAMP complex competes with the cAMP produced by the cells for binding to the cAMP-specific antibody. EA then forms any unbound ED-cAMP and active β-Gal. This active enzyme then converts the chemiluminescent substrate to a standard An output signal is generated that is recorded in a microplate reader.

[0177] Briefly, 10,000 cells per well were plated overnight and then each well was The wells (cells incubated with 10 μL assay buffer) were incubated at 37°C for 30 min. During the incubation, 4× serial concentrations of test compound and cAMP antigen in cell assay buffer (5 μL) were added. Incubate with the enzyme reagent (5 μL). Contains transporter substrate (Emerald II-Galacton Star, 5:1) Then, 20 μL of cell lysis buffer was added and incubated at room temperature for 60 minutes. Next, add 20 μL of EA β-Gal fragment reagent. Incubate for 120 minutes at room temperature. After incubation, chemiluminescence was measured by a plate reader (Envision). and the data is used to calculate an EC50 value for the test peptide.

[0178] The results are shown in Table 2. [Table 4] TIFF2025004067000026.tif48160

[0179] Preparation of the ionic complexes and pharmaceutical compositions of the present invention [Example 1] Preparation of Formulation 2A (Peptide 1 and mPEG-10,000-monocarboxylate) Mixture of mPEG-10,000-mono-carboxylate (5.4 g) in 8.4 g water The mixture was placed in a vial, sealed, and autoclaved at 121°C for 15 minutes. Upon cooling, the vials were transferred to a sterile flow hood. The vial was opened and a 0.2 μm filter was used to pre-filter peptide 1. Sterile aqueous solution (100 mg in 1 mL) was mixed with it. The formulated peptide was homogenous. A clear, viscous aqueous solution was obtained. The concentration of peptide 1 was assayed by HPLC analysis. was 9.9 mg / mL.

[0180] [Example 2] Preparation of Formulation 2B (Peptide 1 and PEG-10,000-dicarboxylate) A mixture of PEG-10,000-dicarboxylate (2.7 g) in 2.0 g of water was The vials were stoppered, sealed and autoclaved at 121°C for 15 minutes. Upon cooling, the vial was transferred to a sterile flow hood. The vial was opened and diluted with sterile water containing peptide 1, which had been pre-filtered through a 0.2 μm filter. The peptide was mixed with the soluble solution (50 mg in 0.5 mL). The resulting solution was a viscous aqueous solution at 100°C. The concentration of peptide 1 was 1.0, as assayed by HPLC analysis. The concentration was 0.8 mg / mL.

[0181] [Example 3] Preparation of Formulation 2C (Peptide 1 and mPEG-20,000-mono-carboxylate) 3.5 g of propylene glycol-ethanol-water mixture (40:15:45 ratio v / v A mixture of PEG-m20,000-carboxylate (1.8 g) in 100 mL of 10 ... The vials were stoppered, sealed, and autoclaved at 121°C for 15 minutes to obtain a uniform permeability. A clear, viscous aqueous solution was obtained. Upon cooling, the vial was transferred to a sterile flow hood. Open the tube and add a sterile aqueous solution of peptide 1 (0. The formulated peptide was a homogeneous, clear, viscous aqueous solution. The concentration of peptide 1 was 6 mg / mL as assayed by HPLC analysis. there were.

[0182] [Example 4] Preparation of formulation 2D (peptide 1 and sodium DPPA) Sodium DPPA (302 mg) in 8.6 g of a 1:1 propylene glycol-water mixture The mixture was placed in a vial. The vial was capped, sealed, and incubated at 121°C for 15 min. Upon cooling, the vial was transferred to a sterile flask and centrifuged to give a homogenous lipid dispersion. Transfer to raw food. Open the vial and add pre-filtered peptidase 1 (PEP) through a 0.2 μm filter. A sterile aqueous solution of tide 1 (100 mg in 1 mL) was mixed with it. The formulated peptide was The resulting solution was a homogeneous, non-viscous dispersion. The concentration of Tid1 was 9.6 mg / mL.

[0183] [Example 5] Preparation of Formulation 2E (Peptide 1 and Sodium Stearate) A mixture of mannitol (300 mg) and sodium stearate (165 mg) The vial was then placed in 8.3 g of water. The vial was then sealed and heated at 75° C. for 0.5 hours. Upon cooling, about 400 μL of 1.0N acetic acid was added to obtain a clear aqueous solution. The pH was adjusted to within 7. The vials were stoppered, sealed, and incubated at 121° C. for 15 min. Upon cooling, the vials were transferred to a sterile flow hood. Open the tube and add a sterile aqueous solution of peptide 1 (1. The formulated peptide was a homogeneous light milky white liquid. The product was obtained as a white, opaque, non-viscous suspension. The concentration of dopamine 1 was 9.7 mg / mL.

[0184] [Example 6] Formulation #3B (a PEG containing sodium stearate and mPEG-2,000-DSPE) Preparation of peptide 1) mPEG-2000-DSPE (1.85 g) and sodium stearate (100 The mixture (1.1 mg) was placed in 10.8 mL of water in a vial. The vial was capped and 1 The mixture was heated at 75°C for 2 h. Upon cooling, approximately 240 μL of acetic acid was added to give a concentration of 6-7 The vial was stoppered, sealed, and incubated at 121°C for 15 min. Upon cooling, the vials were transferred to a sterile flow hood. , a sterile aqueous solution of peptide 1 (1 in 1.2 g) pre-filtered through a 0.2 μm filter The formulated peptide was a uniform light milky opaque non-transparent liquid. A viscous suspension was obtained. The concentration of peptide 1 was 10 The concentration was .1mg / mL.

[0185] [Example 7] Formulation #3C (Peptide 1 and Sodium DPPA and PEG-10,000-dicarboxylic Acid Preparation of carboxylate PEG-10,000-dicarboxylate (1.68 g) and sodium DPPA The mixture (362.4 mg) was placed in 9.1 mL of water in a vial. The vial was capped. The flask was sealed and autoclaved at 121°C for 15 minutes to obtain a homogeneous, milky, opaque suspension. Upon cooling, the vials were transferred to a sterile flow hood. The vials were opened and A sterile aqueous solution of peptide 1 (12 in 1.2 g) pre-filtered through a 0.2 μm filter The formulated peptide was mixed with 100 mg of 100% ethanol. The concentration of peptide 1 was 10.6 mg / mL as assayed by HPLC analysis. there were.

[0186] [Example 8] Preparation of Formulation #3D (Peptide 1 and Sodium DPPA and PEG-3,350) PEG-3,350 (1.8 g contained in 10.8 mL aqueous solution) and sodium The mixture was placed in a vial. The vial was capped and sealed. Autoclaving at 121°C for 15 minutes resulted in a homogeneous milky opaque suspension. Upon cooling, the vials were transferred to a sterile flow hood. The vials were opened and 0.2 μm A prefiltered sterile aqueous solution of peptide 1 (120 mg in 1.2 g) was The formulated peptide was obtained as a homogeneous, milky, opaque suspension. The concentration of peptide 1 was 10.1 mg / mL as assayed by PLC analysis.

[0187] [Example 9] Formulation #3E (peptide 1 and sodium DPPA, PEG-3,350, and sodium Preparation of sodium carboxymethyl ester (CMC) Aqueous solution of sodium CMC (Av MW 90,000) (72 mg in 9.0 mL) was mixed with PEG-3,350 (1.8 g). The resulting clear aqueous solution To the flask was added DPPA (216 mg). The vial was stoppered, sealed, and heated at 121 °C for 15 min. Autoclaving for 1 min resulted in a homogeneous milky opaque suspension. The vial was then transferred to a sterile flow hood. The vial was opened and filtered through a 0.2 μm filter. A previously filtered sterile aqueous solution of peptide 1 (120 mg in 1.2 g) was mixed with it. The formulated peptide was obtained as a homogeneous, milky, opaque suspension. When assayed again, the concentration of peptide 1 was 10.9 mg / mL.

[0188] [Example 10] Formulation #3F (Peptide 1 and mPEG-2,000-DSPE and sodium CM C) Preparation Sodium CMC (Av MW 90,000) (72 mg in 10.8 mL or 6. An aqueous solution of mPEG-2,000-DSPE (7 mg / mL) was mixed with mPEG-2,000-DSPE (1.85 g). The vials were stoppered, sealed, and autoclaved at 121°C for 15 minutes to obtain a homogeneous, clear solution. Upon cooling, the vial was transferred to a sterile flow hood. In the vial containing the mixture of thorium CMC and mPEG-2,000-DSPE The contents were mixed overnight resulting in a clear aqueous solution which was sterile filtered through a 0.2 μm filter. The vials were opened inside a sterile flow hood and prefiltered with a 0.2 μm filter. A filtered sterile aqueous solution of Peptide 1 (120 mg in 1.2 g) was mixed with it. The peptide was obtained as a homogeneous, clear aqueous solution. The concentration of peptide 1 was 10.5 mg / mL.

[0189] [Example 11] Preparation of Formulation #4B (Peptide 1 and mPEG-2,000-DSPE) mPEG-2,000-DSPE (1.855 g) and mannitol (181 mg) The mixture was taken up in 8.8 mL of water for injection (purged with nitrogen for 10 minutes) in a vial. The flask was stoppered and gently swirled to thoroughly wet all solid ingredients with water. The contents of the vial at this stage were then autoclaved at 121°C for 15 minutes. The solution was clear and aqueous. Upon cooling, the vial was transferred to a sterile flow hood. Open and separate the sterile aqueous solution of peptide 1 (1.2) pre-filtered through a 0.2 µm filter. The peptide was mixed with 120 mg of 1000 mL ... The concentration of peptide 1 was 8.7 mg / mL as assayed by HPLC analysis. It was.

[0190] [Example 12] Preparation of Formulation #4C (Peptide 1 and mPEG-2,000-DSPE and CMC) An aqueous solution of sodium CMC (Av MW 90,000) at a concentration of 8.9 mg / mL was prepared in nitrogen-purged water for injection. 9.8 mL of this solution was placed in a vial. , mPEG-2,000-DSPE (1.853 g) and mannitol (182 mg) The vial was capped and gently swirled to ensure that all solid components were thoroughly mixed with the solvent. It was then autoclaved at 121°C for 15 minutes. The contents of the vial were a clear aqueous solution. During cooling, the vial was placed in a sterile flow hood. The vial was opened and the peptide 1, pre-filtered through a 0.2 μm filter, was added to the flask. The bacterial aqueous solution (120 mg in 1.2 g) was mixed with it for 1 hour. A homogeneous, clear aqueous solution was obtained. The concentration of peptide 1 was assayed by HPLC analysis. was 8.0 mg / mL.

[0191] [Example 13] Preparation of Formulation #4D (Peptide 1 and mPEG-2,000-DSPE and DPPA) Manufacturing mPEG-2,000-DSPE (1.86 g), sodium DPPA (363 mg) A mixture of mannitol (60.2 mg) and 9 mL of water for injection (10 mL) was added to a vial. The vial was capped, gently swirled, and completely washed with water. The solid components were then completely wetted. They were then autoclaved at 121° C. for 15 minutes. The contents of the vial at this stage were a light milky homogenous suspension. The vial was then transferred to a sterile flow hood. The vial was opened and filtered through a 0.2 μm filter. A pre-filtered sterile aqueous solution of peptide 1 (120 mg in 1.2 g) was added to it for 1 h. Upon mixing, a homogeneous, light milky, opaque, non-viscous suspension was obtained. When assayed, the concentration of peptide 1 was 9.2 mg / mL.

[0192] [Example 14] Preparation of Formulation #4E (Peptide 1 and mPEG-2,000-DSPE and CMC) An aqueous solution of sodium CMC at a concentration of 7.2 mg / mL was prepared in nitrogen purged water for injection. The solution was taken in a vial and 10 mL of the solution was added to the solution. PE (0.94 g) and mannitol (181.7 mg) were added. The vial was capped. The mixture was then gently swirled to thoroughly wet all solid components with the solvent. Autoclave at 121°C for 15 minutes. The contents of the vial at this stage were clear and sterile. Upon cooling, the vial was transferred to a sterile flow hood. Open and pre-filter a sterile aqueous solution of peptide 1 (1.2 g The peptide was mixed with 120 mg of ethanol for 1 hour. The peptide was formulated as a homogeneous, clear aqueous solution. The concentration of peptide 1 was 9.6 mg / mL as assayed by HPLC analysis. there were.

[0193] [Example 15] Preparation of Formulation #4F (Peptide 1 and mPEG-2,000-DSPE and DPPA) Manufacturing mPEG-2,000-DSPE (0.954 g), sodium DPPA (183.7 A mixture of 61.7 mg of mannitol and 9.6 mL of injection solution was placed in a vial. Water (10 min, nitrogen purged). The vial was capped, gently swirled, and then soaked in water. It was then autoclaved at 121°C for 15 minutes. The contents of the vial at this stage was a light milky homogenous suspension. Upon cooling, the vials were transferred to a sterile flow hood. The vials were opened and filtered using a 0.2 μm filter. A sterile aqueous solution of peptide 1 (120 mg in 1.2 g) prefiltered through a filter was The mixture was mixed for 1 hour to give a homogeneous, light milky, opaque, non-viscous suspension. When assayed by precipitation, the concentration of peptide 1 was 9.9 mg / mL.

[0194] [Example 16] Peptide 1, mPEG-2,000-DSPE, CMA, and D-mannitol Preparation of formulations [Table 5]

[0195] The following preparations are based on a 10 mL formulation to provide the 1 mL final volume in the table above. Can be easily tailored.

[0196] Sodium CMC (Av MW 90,000) (80 mg) and D-mannitol (220 mg) in a pre-weighed vial with a stir bar and stopper. Into this solution, solid sodium mPEG-2,000- Add DSPE (1 g). Stopper and seal the vial. Place it in a 60°C flask. Place in a water bath and stir the contents. Alternatively, the vial is maintained at 60°C. The contents of the vial may be cloudy at first, but The solution will turn into a clear, homogenous aqueous solution within 1-2 hours. Allow the vial to come to room temperature. Then, open it and extract peptide 1 (100 mg in 1 mL, based on its net peptide content) ) into the water. The contents may become cloudy for a moment, but will become clear as soon as they are mixed. The solution will turn into a homogenous aqueous solution. Weigh the vial and determine the weight of the total content required. The resulting solution is adjusted to 10.1 gm by adding an amount of water for injection. The resulting formulation is mixed and filtered through a 0.2 μm filter inside a sterile flow hood. The peptide was obtained as a homogeneous, clear aqueous solution and assayed by HPLC. Verify that the peptide-1 is at a concentration of 10 mg / ml.

[0197] [Example 17] Preparation of formulations with peptide 1, sodium DPPA, and PEG-3,350 [Table 6]

[0198] The following preparations are based on a 10 mL formulation to provide the 1 mL final volume in the table above. Can be easily tailored.

[0199] In a pre-weighed stoppered vial, add 1.2 gm of PEG-3350 and 8 gm of Add 1 mL of water for injection. The contents are dissolved by mixing and contain 302 mg of sodium D Add PPA. Stopper the vial, seal it, and place it in an autoclave at 121°C for 1 hour. Autoclaving for 5 minutes resulted in a homogeneous, milky, opaque suspension. Transfer the vial to a sterile flow hood. Open the vial and filter with a 0.2 μm filter. Prepare a pre-filtered sterile aqueous solution of peptide 1 (100 mg in 1 mL water, the pure peptide Mix the total contents of the vial with an appropriate amount of Water for Injection. Adjust to 10.1 gm by adding additional. Upon mixing, the formulated peptide will become a uniform milky white. The purified peptide was assayed by HPLC to obtain Verify that the peptide-1 is at a concentration of 10 mg / ml.

[0200] [Example 18] Administration of formulations to cynomolgus monkeys to assess pharmacokinetics Groups of cynomolgus monkeys (average weight range 3-7 kg) were cultured as described above. Groups of 6 were randomized for formulation 1 and formulation 3A. Each monkey was administered a single bolus dose of the formulation, calculated at 0.5 mg / kg body weight, into the shoulder. Blood samples were taken starting 30 minutes after dosing and at 36 or 48 hours. Plasma was collected and analyzed using LC-MS / MS technology for peptide analysis. The pharmacokinetic data are shown in Table 3 below. The objective profiles are illustrated in Figures 1, 2, and 3. [Table 7] TIFF2025004067000030.tif107147

[0201] The results of a comparative pharmacokinetic study with ionic complexes of peptide-1 demonstrated Thus, there was a significant decrease in C-max (1100 ng / mL of peptide 1 Cmax vs. 228 ng / mL observed with conjugate 3D. The carboxylic conjugates showed weaker C-max in a similar range compared to peptide-1. In addition, T-max was also significantly increased with some of these ionic complexes (many The ionic complex had a T-max of several hours, whereas the T-max of peptide-1 was was 0.5 hours).

[0202] The teachings of all patents, published applications, and references cited herein are incorporated by reference in their entirety. No. 6,399,433, filed on Oct. 23, 2003, and are incorporated by reference in their entirety.

[0203] While the present invention has been particularly shown and described with reference to illustrative embodiments thereof, the appended claims are intended to provide a further understanding of the invention as claimed. No variations in form and detail may be made without departing from the scope of the invention as encompassed by the claims. It will be understood by those skilled in the art that various changes may be made therein.

Claims

1. Cationic polypeptides and PEG-carboxylic acids, lipids having 10 or more carbon atoms Ionic complexes comprising an anionic excipient selected from acids, phospholipids, and combinations thereof body.

2. The molar ratio of the cationic peptide to the anionic excipient ranges from about 1:1 to about 1:

10. The ionic complex of claim 1 .

3. The anionic excipient is lauric acid, myristic acid, palmitic acid, stearic acid, Lachysinic acid, myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid acid, vaccenic acid, linoleic acid, linoelaidic acid, α-linolenic acid, arachidonic acid, 2. The fatty acid of claim 1, wherein the fatty acid is selected from the group consisting of cosapentaenoic acid, ... and phytanic acid.

3. The ionic complex according to any one of claims 1 to 2.

4. The ionic complex of claim 3, wherein the fatty acid is stearic acid.

5. The anionic excipient according to any one of claims 1 to 2, wherein the anionic excipient is a phospholipid. sexual complex.

6. The phospholipid is L-α-phosphatidic acid, 1-oleoyl lysophosphatidic acid, L-α-phosphatidylglycerol, 1,2-di-O-tetradecyl-sn-glycerol -3-phospho-(1'-rac-glycerol), 1,2-dimyristoyl-sn-glycerol Cero-3-phospho-(1'-rac-glycerol), 1,2-dimyristoyl-sn- Glycerol-3-phospho-L-serine, 1,2-dipalmitoyl-sn-glycero-3-phospho Phosphatidic acid, 1,2-distearoyl-sn-glycero-3-phosphatidic acid, m PEG-2,000-DSPE, mPEG-5,000-DSPE, 1-(1,2-dihe (Xadecanoylphosphatidyl)inositol-4,5-bisphosphate, trisodium salt, and 1-(1,2-dihexadecanoylphosphatidyl)inositol-3,4 , 5-triphosphate, tetrasodium salt, 1-palmitoyl-2-oleoyl phosphat Tidylglycerol, 1-palmitoyl-2-arachidonoyl-sn-glycero-3-hydroxy Sufoglycerol, DSPG, DPPG, DEPG, DOPG, DEPA, DOPA, D SPS, DPPS, DEPS, DOPS, L-α-lysophosphatidylserine, L-α- Lysophosphatidylinositol, Tetradecylphosphonic acid, L-α-phosphatidylinositol Inositol-4-phosphate, L-α-phosphatidylinositol-4,5-bisphosphatase 1,2-diphytanoyl-sn-glycero-3-phosphate, -O-tetradecyl-sn-glycero-3-phospho-(1'-rac-glycerol), mPEG-2,000-DSPE, mPEG-5,000-DSPE, and mixtures thereof The ionic complex according to claim 5 , wherein the ionic complex is selected from the group consisting of

7. The phospholipid is L-α-phosphatidic acid, 1-oleoyl lysophosphatidic acid, Tetradecylphosphonic acid, L-α-phosphatidylglycerol, L-α-phosphatidyl Ruinositol, L-α-phosphatidylserine, 1,2-di-O-tetradecyl-sn -glycero-3-phospho-(1'-rac-glycerol), 1,2-dimyristoyl- sn-Glycero-3-phospho-(1'-rac-glycerol), 1,2-dimyristyl 1,2-distearoyl-sn-glycero-3-phospho-L-serine, 3-phospho-L-serine, distearoylphosphatidylglycerol, dipalmitoyl 1,2-dipalmitoyl-sn-glycero-3-phosphatidylglycerol, Phatidic acid (DPPA), 1,2-distearoyl-sn-glycero-3-phosphatidylinositol PEG-2,000-DSPE, mPEG-5,000-DSPE, 1-(1 , 2-dihexadecanoylphosphatidyl)inositol-4,5-bisphosphate, Trisodium salt, and 1-(1,2-dihexadecanoylphosphatidyl)inositol tetrasodium salt, tetrahydrofuran salt, and mixtures thereof. The pharmaceutical composition according to claim 5 .

8. The anionic lipid is 1,2-dipalmitoyl-sn-glycero-3-phosphatidine The ionic complex of claim 5 which is an acid.

9. The anionic lipid is mPEG-2,000-DSPE, and DSPE is 1,2- The compound according to claim 5, which is distearoyl-sn-glycero-3-phosphoethanolamine. Ionic complexes of.

10. The method according to any one of claims 1 to 2, wherein the anionic excipient is a PEG-carboxylic acid. Ionic complexes as described above.

11. The PEG-carboxylic acid may be PEG-10,000-monocarboxylate, PEG- 20,000-monocarboxylate, mPEG 1,000-monocarboxylate, m PEG-2,000-monocarboxylate, mPEG-5,000-monocarboxylate mPEG-10,000-monocarboxylate, mPEG-20,000-monocarboxylate Carboxylate, mPEG-30,000-monocarboxylate, mPEG-40, PEG-1,000-monocarboxylate, PEG-1,000-dicarboxylate, PEG-2 ,000-dicarboxylate, PEG-3,500-dicarboxylate, PEG-5 ,000-dicarboxylate, PEG-7,500-dicarboxylate, PEG-1 PEG-40,000-dicarboxylate, and Y-shaped PEG-40,000-monocarboxylate 11. The ionic complex of claim 10, wherein the ionic complex is selected from the group consisting of aryl esters.

12. The PEG-carboxylic acid is PEG-5000-monocarboxylate, PEG-10 ,000-monocarboxylate, PEG-20,000-monocarboxylate, mP EG-5000-monocarboxylate, mPEG-10,000-monocarboxylate mPEG-20,000-monocarboxylate, PEG-5000-dicarboxylate PEG-10,000-dicarboxylate, and PEG-10,000-dicarboxylate. The ionic complex according to claim 1.

13. 10. The ionic complex of claim 1 comprising a combination of anionic excipients.

14. The combination of anionic excipients is stearic acid and mPEG-2,000-DS. PE, DPPA and PEG-10,000-dicarboxylate, DPPA and mP EG-2,000-DSPE, mPEG-2,000-DSPE and CMC, and 14. The ionic complex of claim 13, selected from mPEG-2,000-DSPE.

15. The cationic polypeptide has one, two, three, four, five, six, or more positive charges. The ionic complex according to claim 1 .

16. The cationic polypeptide has the structural formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, 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, respectively, are independently H or C1-C6 alkyl; _ １ は、; , Tyr, Ala, Ser, Thr, Gln, Asn, Asp, Glu, or TzA is an amino acid residue selected from A １ is absent or A １ is an optionally substituted C1-C12 alkyl, an optionally substituted C6-C 18 aryl, optionally substituted C5-C18 heteroaryl, and aralkyl The aryl moiety is an optionally substituted C6-C18 aryl. The alkyl portion is optionally substituted C1-C12 alkyl or heteroaralkyl. and the heteroaryl moiety is an optionally substituted C5-C18 heteroaryl. and the alkyl portion is an optionally substituted C1-C12 alkyl; A ２ and A ８ are each independently Cys, hCys, Pen, Asp, Glu , Lys, Orn, Dbu, or Dpr; ２ Oh B.A. ８ are chosen in pairs so that they can form covalent bonds between their respective side chains. Selected, A ３ We are here to help you, we ... , Lys, Arg, His, Phe, Gln, Sar, Gly, Asn, or Aib or absent, A ４ is His, Atc, Ala, QAla, which may be absent or substituted; Aib, Sar, Ser, Thr, Pro, Hyp, Asn, Gln, Trp, Tyr, an amino acid residue selected from Lys, Arg, sChp, or residue X, wherein X is An amino acid represented by the following structural formula: 【Chemistry 2】 A ５ is optionally substituted Phe, optionally substituted 1-Nal, or 2-Nal which is optionally substituted; A ６ is Arg, A ７ is Trp, Any amino acid residue may be in the L- or D-configuration; However, A ３ and A ４ provided that both are not absent. The ionic complex of claim 1 represented by formula (I):

17. A ３ The ionic complex of claim 16, wherein is a D-amino acid.

18. A ４ The ionic complex of claim 16, wherein is an L-amino acid.

19. A ４ The ionic complex of claim 16, wherein is absent.

20. A ５ is optionally substituted D-Phe, The ionic complexes described herein.

21. A ５ may be substituted at any of the five aromatic carbons, the substituents being F, CI, Br, I, -CH ３ , -OH, -CN, amine, -NO ２ , or -OCH ３ Selected from The ionic complex according to any one of claims 16 to 20.

22. A ５ が、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'-ジ-CH ３ )、Phe(2',4'-ジ-OCH ３ ()、Phe (3',4'-diF), Phe(3',4'-diCl), Phe(3',4'-diBr) , Phe (3',4'-di-l), Phe (3',4'-di-CN), Phe (3',4' -zi-CH ３ ), Phe(3',4'-di-OCH ３ ), Phe(3',5'-diF), Phe (3',5'-diCl), Phe (3',5'-diBr), Phe (3',5'- di-l), Phe(3',5'-di-CN), Phe(3',5'-diCH ３ ), Phe( 3',5'-di-OCH ３ ), or Phe(3',4',5'-triF) The ionic complex according to any one of claims 16 to 20, wherein the D-amino acid residue is

23. A ４ is His optionally substituted at any substitutable position, and the substituents are F, CI, Br, I, -CH ３ , -OH, -CN, amine, -NO ２ , or -OCH ３ from The ionic surfactant according to any one of claims 16 to 18 and claims 20 to 22 is selected from the group consisting of ionic surfactants, Complex.

24. The cationic polypeptide is represented by the following structural formula: 【Chemistry 3】 or a pharma- ceutically acceptable salt thereof.

25. The cationic polypeptide is represented by the following structural formula: 【Chemistry 4】 【change】 or a pharma- ceutically acceptable salt thereof.

26. The cationic polypeptide is represented by the following structural formula: 【Chemistry 5】 or a pharma- ceutically acceptable salt thereof.

27. The cationic polypeptide is represented by the following structural formula: 【Chemistry 6】 or a pharma- ceutically acceptable salt thereof.

28. A ４ To see, to ... p, Asn, Gln, substituted His, Trp, Tyr, Lys, Arg, sChp, or 17. The ionic complex of claim 16, wherein residue X is an amino acid residue selected from:

29. The cationic polypeptide has any one of the following structural formulas: Ac-Arg-cyclo[Cys-D-Ala-His(3-Me)-D-Phe-Ar g-Trp-Cys]-NH ２ (SEQ ID NO: 6), Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Ar g-Trp-Cys]-NH ２ (SEQ ID NO: 7), Ac-Arg-cyclo[Cys-D-Ala-Trp-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 8), Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 9), Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 10), Ac-Arg-cyclo[Cys-D-Ala-Arg-D-Phe-Arg-Trp- Cys]-NH (SEQ ID NO: 11), Ac-Arg-cyclo[Cys-D-Ala-Tyr-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 12), Ac-Arg-cyclo[Cys-D-Ala-D-Pro-D-Phe-Arg-Tr p-Cys]-NH ２ (Allocation number: 13), Ac-Arg-Cyclo[Cys-D-Ala-Pro-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 14), Ac-Arg-cyclo[Cys-D-Ala-Pro-D-Phe(p-F)-Arg -Trp-Cys]-NH ２ (SEQ ID NO: 15), Ac-Arg-cyclo[Cys-D-Ala-Atc-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 16), Ac-Arg-cyclo[Cys-D-Ala-QAla-D-Phe-Arg-Trp -Cys]-NH ２ (SEQ ID NO: 17), Ac-Arg-cyclo[Cys-D-Ala-sChp-D-Phe-Arg-Trp -Cys]-NH ２ (SEQ ID NO:18), or Ac-Arg-cyclo[Cys-D-Ala-X-D-Phe-Arg-Trp-Cy s]-NH ２ (SEQ ID NO: 19) or a pharma- ceutically acceptable salt thereof. Merge.

30. The cationic polypeptide has any one of the following structural formulas: Ac-Arg-cyclo[hCys-Ala-D-Phe-Arg-Trp-Cys]- N.H. ２ (SEQ ID NO: 20), Ac-Arg-cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Cys ]-NH ２ (SEQ ID NO: 21), Ac-Arg-cyclo[hCys-D-Ala-D-Phe-Arg-Trp-Pen ]-NH ２ (SEQ ID NO: 22), Ac-Arg-cyclo[Glu-D-Ala-D-Phe-Arg-Trp-Dpr] -NH ２ (SEQ ID NO: 23), Ac-Arg-cyclo[Glu-Ala-D-Phe-Arg-Trp-Dpr]-N H ２ (SEQ ID NO: 24), Ac-Arg-cyclo[hCys-Aib-D-Phe-Arg-Trp-Cys]- N.H. ２ (SEQ ID NO: 25), Ac-Arg-cyclo[hCys-Sar-D-Phe-Arg-Trp-Cys]- N.H. ２ (SEQ ID NO: 26), Ac-Arg-cyclo[hCys-Val-D-Phe-Arg-Trp-Cys]- N.H. ２ (SEQ ID NO: 27), Ac-Arg-cyclo[hCys-D-Val-D-Phe-Arg-Trp-Cys ]-NH ２ (SEQ ID NO: 28), Ac-Arg-cyclo[hCys-Gln-D-Phe-Arg-Trp-Cys]- N.H. ２ (SEQ ID NO: 29), Ac-Arg-cyclo[hCys-D-Gln-D-Phe-Arg-Trp-Cys ]-NH ２ (SEQ ID NO: 30), Ac-Arg-cyclo[hCys-Ala-D-Phe-Arg-Trp-Pen]- N.H. ２ (SEQ ID NO: 31), Ac-Arg-cyclo[D-Pen-D-Ala-D-Phe-Arg-Trp-hC ys]-NH ２ (SEQ ID NO: 32), Ac-Arg-cyclo[Cys-D-Ala-D-Phe-Arg-Trp-hCys ]-NH ２ (SEQ ID NO: 33), Ac-Arg-cyclo[Pen-D-Ala-D-Phe-Arg-Trp-hCys ]-NH ２ (SEQ ID NO: 34), Ac-Arg-cyclo[D-hCys-D-Ala-D-Phe-Arg-Trp-C ys]-NH ２ (SEQ ID NO: 35), Ac-Arg-cyclo[hCys-Pro-D-Phe-Arg-Trp-Cys]- N.H. ２ ,or Ac-Arg-cyclo[hCys-D-Pro-D-Phe-Arg-Trp-Cys ]-NH ２ (SEQ ID NO:37) or a pharma- ceutically acceptable salt thereof. Merge.

31. A ３ To see, to ... , Arg, His, Phe, Gln, Sar, Gly, Asn, or Aib is an amino acid residue that A ４ To see, to ... p, Asn, Gln, substituted His, Trp, Tyr, Lys, Arg, sChp, or 17. The ionic complex of claim 16, wherein residue X is an amino acid residue selected from:

32. The cationic polypeptide has any one of the following structural formulas: Ac-Arg-cyclo[Cys-Val-Gln-D-Phe-Arg-Trp-Cy s]-NH ２ (SEQ ID NO: 38), Ac-Arg-cyclo[Cys-D-Val-Gln-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO:39), or Ac-Arg-cyclo[Cys-D-Val-His(1-Me)-D-Phe-Ar g-Trp-Cys]-NH ２ (SEQ ID NO:40) or a pharma- ceutically acceptable salt thereof. Merge.

33. The cationic polypeptide has any one of the following structural formulas: Ac-TzAla-cyclo[Cys-Ala-Gln-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO:41) or Ac-Glu-cyclo[Cys-Ala-His-D-Phe-Arg-Trp-Cy s]-NH ２ (SEQ ID NO:42) or a pharma- ceutically acceptable salt thereof. Merge.

34. The cationic polypeptide has any one of the following structural formulas: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Ar g-Trp-Cys]-NH ２ (SEQ ID NO:7) Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO:9) Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO: 10), or a pharma- ceutically acceptable salt thereof. Merge.

35. The cationic polypeptide has any one of the following structural formulas: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-2-Nal- Arg-Trp-Cys]-NH ２ (SEQ ID NO: 84), Ac-Arg-cyclo[Cys-D-Ala-Gln-D-2-Nal-Arg-Tr p-Cys]-NH2 (SEQ ID NO:85), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-2-Nal-Arg-Tr p-Cys]-NH2 (SEQ ID NO:86) or a pharma- ceutically acceptable salt thereof. Merge.

36. The cationic polypeptide has any one of the following structural formulas: Ac-Arg-cyclo[Cys-D-Ala-His(1-Me)-D-Phe-Ar g-Trp-Cys]-OH (SEQ ID NO: 87), Ac-Arg-cyclo[Cys-D-Ala-Gln-D-Phe-Arg-Trp- Cys]-OH (SEQ ID NO:88), or Ac-Arg-cyclo[Cys-D-Ala-Asn-D-Phe-Arg-Trp- Cys]-OH (SEQ ID NO:89) or a pharma- ceutically acceptable salt thereof. Merge.

37. The cationic polypeptide has any one of the following structural formulas: Ac-Arg-cyclo[Cys-D-Leu-His-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO:2), Ac-Arg-cyclo[Cys-D-Ile-His-D-Phe-Arg-Trp- Cys]-NH2 (SEQ ID NO:3), Ac-Arg-cyclo[Cys-D-Tle-His-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO:4), or Ac-Arg-cyclo[Cys-D-Val-His-D-Phe-Arg-Trp- Cys]-NH ２ (SEQ ID NO:5) or a pharma- ceutically acceptable salt thereof. body.

38. The cationic polypeptide is selected from the group consisting of leuprolide, buserelin, histrelin, and gosserelin. , deslorelin, nafarelin, triptorelin, aborelin, abarelix, seto Lorelix, degarelix, ganirelix, octreotide, lanreotide, vapreviral amide, pasireotide, glucagon, amylin, pramlintide, insulin, glucagon-like GLP-1, GLP-1 agonist, human growth hormone, Exenatti parathyroid hormone, adrenocorticotropic hormone, botulinum toxin, amyloid peptide, Cholecystokinin, calcitonin, conotoxin, prialt, gastric inhibitory peptide, in Thyroid-like growth factor, growth hormone releasing factor, antibacterial factor, glatiramer, Hematide , nesiritide, ANF peptide, angiotensin peptide, ACTH, melanocortin , opioid peptides, dynorphins, oxytocin, oxytocin analogs, vasopressin Resin, vasopressin analogs, somatostatin, and somatostatin analogs The ionic complex according to claim 1 , wherein the ionic complex is selected from the group consisting of:

39. The ionic complex according to any one of claims 1 to 38 and a pharma- ceutically acceptable salt thereof A pharmaceutical composition comprising a carrier.

40. The pharma- ceutically acceptable carrier may be PEG, polyol, ethanol, DMSO, Selected from NMP, DMF, benzyl alcohol, water, a pH stable buffer solution, and mixtures thereof.

40. The pharmaceutical composition of claim 39,

41. The pharma- ceutically acceptable carrier is PEG with an average molecular weight of 100 to 5,000. The pharmaceutical composition of claim 40.

42. The polyol is propylene glycol, tripropylene glycol, glycerol 41. The pharmaceutical composition of claim 40, wherein said compound is selected from the group consisting of:

43. The concentration of the cationic polypeptide is in the range of about 0.01 mg / mL to about 100 mg / mL. The pharmaceutical composition of claim 39.

44. 4. The method of claim 3, wherein the concentration of the cationic polypeptide is from about 1 mg / mL to about 50 mg / mL.

4. The pharmaceutical composition described in 3.

45. a molar ratio based on the charge of the cationic polypeptide to the charge of the anionic excipient , about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8 , about 1:9, or 1:

10. Pharmaceutical compositions.

46. The ionic complex precipitates in a physiological environment to form a drug depot. Item 46. The pharmaceutical composition according to any one of Items 39 to 45.

47. 47. Any of claims 39 to 46, wherein the composition comprises at least one further excipient. The pharmaceutical composition described in claim 1.

48. The additional excipients may be a pH stable buffer, a preservative, a surfactant, a stabilizer, an antioxidant, 48. The composition of claim 47, wherein the polymer is selected from the group consisting of a tonicity agent, a tonicity agent, a non-ionic polymer, and an ionic polymer. The pharmaceutical composition described.

49. The cationic polypeptide of the ionic complex according to any one of claims 1 to 38 has A method for treating a subject suffering from a disease or disorder responsive to pharmacological activity. The method includes administering to a subject a therapeutically effective amount of an ionic complex according to claims 1 to 38. How to do it.

50. 50. The method of claim 49, wherein the disease or disorder is responsive to modulation of an MCR receptor. 。

51. The disease or disorder is selected from the group consisting of type 1 diabetes, type 2 diabetes, obesity, insulin resistance, and metastasis.

51. The method of claim 50, selected from the treatment of chronic obstructive pulmonary disease.

52. Cationic polypeptides and PEG-carboxylic acids, lipids having 10 or more carbon atoms Ionic complexes comprising an anionic excipient selected from acids, phospholipids, and combinations thereof 1. A method for producing a body, comprising: a) preparing a mixture of said anionic excipient and an aqueous excipient diluent; b) autoclaving the mixture under conditions sufficient to sterilize the excipients. P, c) providing a sterile peptide solution comprising the cationic polypeptide and an aqueous peptide diluent; adding to the excipient mixture.

53. 53. The method of claim 52, wherein the excipient mixture is a suspension.

54. 53. The method of claim 52, wherein the excipient mixture is an aqueous solution.

55. Cationic polypeptides and PEG-carboxylic acids, lipids having 10 or more carbon atoms Ionic complexes comprising an anionic excipient selected from acids, phospholipids, and combinations thereof 1. A method for producing a body, comprising: a) preparing an aqueous solution of said anionic excipient and aqueous excipient diluent; b) filtering the aqueous solution of step a through a 0.2 micron filter; c) sequestering the sterile peptide solution containing the cationic polypeptide and an aqueous peptide diluent; The method comprises adding to the aqueous excipient solution of step b.

56. Cationic polypeptides and PEG-carboxylic acids, lipids having 10 or more carbon atoms Ionic complexes comprising an anionic excipient selected from acids, phospholipids, and combinations thereof 1. A method for producing a body, comprising: a) a composition comprising the anionic excipient, an aqueous excipient diluent, and the cationic polypeptide; preparing an aqueous solution; b) Sterilize the resulting aqueous solution by filtering through a 0.2 micron filter. The method comprises the step of sterilizing.

Images