Formulations for non-naturally occurring melanocortin receptor analogs for modulating weight gain

Abstract

Provided herein are stable aqueous pharmaceutical compositions and formulation comprising non-naturally occurring melanocortin antagonist analogs. The non-naturally occurring melanocortin antagonist analog comprises a sequence of Formula (I) Ac-Nle-c[Asp-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond.

Description

INJECTABLE FORMULATIONS FOR NON-NATURALLY OCCURRING MELANOCORTIN RECEPTOR ANTAGONIST ANALOGS FOR MODULATING WEIGHT GAINCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 704,513, filed October 7, 2024, which is incorporated herein by reference in its entirety.INCORPORATION BY REFERENCE OF SEQUENCE LISTING

[0002] This application contains an ST.26 compliant Sequence Listing, which is submitted concurrently in xml format and is hereby incorporated by reference in its entirety. The .xml copy, created on October 3, 2025 is titled “183B-412934-WO_SL.xml” and is 4,498 bytes in size.BACKGROUND

[0003] A major challenge in the development of peptide-based therapeutics formulations is maintaining solubility, stability, and potency of the peptide. It is particularly desirable to avoid aggregates and particulates in injectable formulations that require sterile filtration before administration (e.g., parenteral injection). Further, the formation of aggregates in injectable formulations may cause adverse reactions at the injection site.

[0004] Melanocortin analogs have been synthesized for the potential regulation and treatment of many conditions, including weight regulation (e.g., obesity, anorexia, and cachexia), hormonal secretion, and hyposecretion of many exocrine glands. Despite recent advances, there is still a need for melanocortin analog-containing compositions that are stable, effective, and suitable for parenteral administration, including subcutaneous injection.SUMMARY

[0005] Provided herein are pharmaceutical compositions having a pH of about 4.0 to about 6.5, comprising a non-naturally occurring melanocortin analog having the sequence:-1 -168785213.5Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), an osmolyte, a pH adjusting agent, and a buffer, wherein c in the sequence represents cyclization via a lactam bond.

[0006] Further provided herein are methods of preparing pharmaceutical compositions and formulations according to the present technology.BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIGS. 1 A-1 F show the total impurities over time in Formulations 1 -8 containing 50 mg (FIGS. 1 A and 1 D), 25 mg (FIGS. 1 B and 1 E), and 12.5 mg (FIGS. 1 C and 1 F) B07 (Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 2) stored at 5 °C (FIGS. 1 A-1 C) and 25 °C (FIGS. 1 D-1 F).DETAILED DESCRIPTION

[0008] The present technology provides stable aqueous pharmaceutical compositions of non-naturally occurring melanocortin analogs. Pharmaceutical compositions described herein have a pH of less than 7.0 and include a non-naturally occurring melanocortin analog and one or more pharmaceutically acceptable carriers and / or excipients. In some embodiments, the pharmaceutical compositions have a pH of about 4.0 to about 6.5.

[0009] Pharmaceutical compositions comprising non-naturally occurring melanocortin analogs of the present technology may be administered to a subject in need thereof to treat and / or prevent conditions associated with weight regulation, such as, for example, cachexia. To achieve desired therapeutic outcomes, the pharmaceutical compositions may be formulated for specific routes of administration, such as parenteral delivery, including, but not limited to, subcutaneous delivery. Accordingly, the one or more pharmaceutically acceptable carriers and / or excipients included in the pharmaceutical compositions described herein may be useful for subcutaneous delivery of the non-naturally occurring melanocortin peptides. In some embodiments, the one or more carriers and / or excipients are selected based on desirable properties, such as reduced or lack of delivery site reactions, solubility, stability (e.g., shelf-life), pH, buffering, pharmacokinetics, bioavailability, or the like, of the pharmaceutical composition. In some embodiments, the one or more pharmaceutically-2-168785213.5acceptable carriers and / or excipients may include a buffer, an osmolyte, and a pH adjusting agent.

[0010] Methods of preparing the stable aqueous pharmaceutical compositions are also described herein. In some embodiments, the pharmaceutical compositions are prepared by combining an aqueous peptide composition with a pharmaceutically acceptable carrier composition.

[0011] The following description is merely exemplary in nature and is not intended to limit the present technology, its applications, or its uses. It should be understood that throughout the drawings, corresponding reference numerals indicate like or corresponding parts and features. The description of specific examples indicated in various embodiments of the present technology are intended for purposes of illustration only and are not intended to limit the scope of the present technology disclosed herein. Moreover, recitation of multiple embodiments having stated features is not intended to exclude other embodiments having additional features or other embodiments incorporating different combinations of the stated features.

[0012] Furthermore, the detailed description of various embodiments herein makes reference to the accompanying drawing / FIGS, which show various embodiments by way of illustration. While the embodiments are described in sufficient detail to enable those skilled in the art to practice the present technology, it should be understood that other embodiments may be realized, and that logical and mechanical changes may be made without departing from the spirit and scope of the present technology. Thus, the detailed description herein is presented for purposes of illustration only and not of limitation. For example, steps or functions recited in any description, method, system, or process, may be executed in any order and are not limited to the order presented. Moreover, any of the steps or functions thereof may be outsourced to or performed by one or more third parties.Definitions

[0013] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the-3-168785213.5present technology belongs. For the purposes of the present technology, the following terms are defined below.

[0014] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0015] The term “about” means a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by acceptable levels in the art. Typically, such variation may be as much 10% above and below a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length and such variation may be influenced by standard applicable measurement practices. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth.

[0016] As used herein, a “composition” or a “pharmaceutical composition” refers to a mixture of the active ingredient with other chemical components, such as pharmaceutically acceptable carriers and / or excipients.

[0017] As used herein, a “pharmaceutically acceptable carrier” refers to a carrier or diluent that does not cause significant irritation to an organism, does not abrogate the biological activity and properties of the administered active ingredient, and / or does not interact in a deleterious manner with the other components of the composition in which it is contained.

[0018] “Melanocortin analogs,” “melanocortin peptides,” or “melanocortins,” are used interchangeably and refer to melanocortin-receptor ligands, which are macromolecules containing at least one melanocortin pharmacophore. Melanocortin analogs are typically peptides that bind melanocortin receptors under physiological conditions. Melanocortin analogs include non-naturally occurring melanocortin peptides and truncated and / or modified versions of melanocortin full-length protein or peptides. For example, the full-length pro-opiomelanocortin protein (POMC), prior to proteolytic cleavage of “sub-peptides,” consists of 241 amino acids. Tissue-specific proteolytic cleavage of POMC yields peptides ranging in size from 13 amino acids to 76 amino acids. See Bicknell and-4-168785213.5Lawry, Encyclopedia of Stress, vol. 3, 257-265, Academic Press (2000). Synthesized, non- naturally occurring melanocortin analogs having increased melanocortin receptor activity as discussed herein are approximately 7-12 amino acids in size. Melanocortin analogs exhibit binding functionality with melanocortin receptors. In addition to peptides, the melanocortin analogs include small molecule analogs of melanocortin or portions thereof comprised of organic compounds, inorganic compounds, or combinations of peptide and small molecule — i.e., peptide mimetics, or various combinations thereof. “Melanocortin peptides” can be structurally similar and / or functionally similar to biological melanocortin proteins in their ability to bind melanocortin receptors. Further, the melanocortin analogs generally contain the pharmacophore: His-Phe-Arg-Trp (SEQ ID NO: 1 ) or a modified version thereof, or a structural or functional peptide mimetic thereof.

[0019] A “pharmacophore” is the minimum set of amino acid residues necessary to achieve a physiological effect; or a small molecule that is (with respect to a receptor) a structural mimic of the amino acid residues required for binding to and activation of a receptor. His-Phe-Arg-Trp (SEQ ID NO: 1 ) and their analogs are the pharmacophore of melanocortin for the regulated physiological effect. Therefore, non-naturally occurring melanocortin pharmacophore analogs can be small peptides or organic molecules designed to mimic the appearance or function (including activation or deactivation of receptor activity) of the melanocortin pharmacophore core sequence peptide.

[0020] A melanocortin receptor “antagonist” or “melanocortin antagonist” is a naturally occurring substance or manufactured drug substance or composition that opposes the melanocortin receptor-associated responses normally induced by a melanocortin receptor agonist agent.

[0021] The terms “bind,” “binding,” “complex,” and “complexing,” refer to all types of physical and chemical binding, reactions, complexing, attraction, chelating and the like.

[0022] The “peptides” described herein can be (a) naturally occurring, (b) produced by chemical synthesis, (c) produced by recombinant DNA technology, (d) produced by biochemical or enzymatic fragmentation of larger molecules, (e) produced by methods resulting from a combination of methods (a) through (d) listed above, or (f) produced by any other means for producing peptides.-5-168785213.5

[0023] The term “peptide” as used herein includes any structure comprised of two or more amino acids, including chemical modifications and derivatives of amino acids. The amino acids forming all or a part of a peptide may be naturally occurring amino acids, stereoisomers and modifications of such amino acids, non-protein amino acids, post- translationally modified amino acids, enzymatically modified amino acids, constructs or structures designed to mimic amino acids, and the like, so that the term “peptide” includes pseudopeptides and peptidomimetics, including structures which have a non-peptidic backbone. The term “peptide” also includes dimers or multimers of peptides. A “manufactured” peptide includes a peptide produced by chemical synthesis, recombinant DNA technology, biochemical, or enzymatic fragmentation of larger molecules, combinations of the foregoing or, in general, made by any other method. The term “peptide” includes peptides containing a variable number of amino acid residues, optionally with nonamino acid residue groups at the N- and C-termini, such groups including acyl, acetyl, alkenyl, alkyl, N-alkyl, amine, or amide groups, among others.

[0024] By employing chemical synthesis, a useful means of production, it is possible to introduce various amino acids which do not naturally occur along the chain, modify the N- or C-terminus, and the like, thereby providing for improved stability and formulation, resistance to protease degradation, and the like.

[0025] “Amino acids” are molecules containing an amine group, a carboxylic acid group, and a side-chain that is specific to each amino acid. The key elements of an amino acid are carbon, hydrogen, oxygen, and nitrogen and have the generic formula H2N — CHR — COOH, wherein R represents a side chain group. The various a-amino acids differ in the side-chain moiety that is attached to the a-carbon. The “amino acids” of the present technology include the known naturally occurring protein amino acids, which are referred to by both their common three letter abbreviation and single letter abbreviation. See generally Synthetic Peptides: A User’s Guide, G. A. Grant, editor, W.H. Freeman & Co., New York (1992), the teachings of which are incorporated herein by reference, including the text and table set forth at pages 11 through 24. As set forth above, the term “amino acid” also includes stereoisomers and modifications of naturally occurring protein amino acids, nonprotein amino acids, post-translationally modified amino acids, enzymatically synthesized-6-168785213.5amino acids, derivatized amino acids, constructs or structures designed to mimic amino acids, and the like. Modified and unusual amino acids are described generally in Synthetic Peptides: A User’s Guide, supra; Hruby et al., Biochem. J. 268:249-262 (1990); and Toniolo, Int. J. Peptide Protein Res. 35:287-300 (1990); the teachings of all of which are incorporated herein by reference.

[0026] The phrase “amino acid side chain moiety” used herein, including as used in the specification and claims, includes any side chain of any amino acid, as the term “amino acid” is defined herein. This thus includes the side chain moiety present in naturally occurring amino acids. It further includes side chain moieties in modified naturally occurring amino acids, such as glycosylated amino acids. It further includes side chain moieties in stereoisomers and modifications of naturally occurring protein amino acids, non-protein amino acids, post-translationally modified amino acids, enzymatically synthesized amino acids, derivatized amino acids, constructs, or structures designed to mimic amino acids, and the like. For example, the side chain moiety of any amino acid disclosed herein is included within the definition. A “derivative” of an amino acid side chain moiety is included within the definition of an amino acid side chain moiety.

[0027] The “derivative” of an amino acid side chain moiety includes any modification to or variation in any amino acid side chain moieties, including a modification of naturally occurring amino acid side chain moieties. By way of example, derivatives of amino acid side chain moieties include straight chain or branched, cyclic or noncyclic, substituted or unsubstituted, saturated or unsaturated, alkyl, aryl or aralkyl moieties.

[0028] In the peptides described herein, conventional amino acid residues have their conventional meaning as given in Chapter 2400, of the Manual of Patent Examining Procedure, 8th Ed. Thus, “Ala” is alanine; “Arg” is arginine; “Asn” is asparagine; “Asp” is aspartic acid; “Cys” is cysteine; “Gin” is glutamine; “Glu” is glutamic acid; “His” is histidine; “lie” is isoleucine; “Leu” is leucine; “Lys” is lysine; “Met” is methionine; “Phe” is phenylalanine; “Pro” is proline; “Ser” is serine; Thr is threonine; “Trp” is tryptophan; “Tyr” is tryosine; and “Vai” is valine. Unless otherwise indicated, all amino acids abbreviations represent either isomer, i.e., the L-isomer, the D-isomer, or combinations thereof can be used. Thus, for example, “L-Phe” or “IPhe” is L-phenylalanine; “D-Phe” or “dPhe” is D--7-168785213.5phenylalanine; dVal is D-valine; dPro is D-proline; “D- / L-Phe” or“d / IPhe” is D-phenylalanine, L-phenylalanine, or combinations thereof; “Phe” is also D-phenylalanine, L-phenylalanine, or combinations thereof, and so on.

[0029] An alpha (a)-amino acid has the generic formula H2N— CaHR— COOH, where R is a side chain moiety and the amino group is attached to the carbon atom immediately adjacent to the carboxylate group (i.e., the oc-carbon). Other types of amino acids exist when the amino group is attached to a different carbon atom, such as, for example p-amino acids and y-amino acids.

[0030] When -amino acids are incorporated into peptides, two main types of p- peptides exist: those with the side chain residue, R, on the carbon next to the amine are called p3peptides and those with the side chain residue on the carbon next to the carbonyl group are called p2amino acids. Further, p-amino acids may adopt L- or D- stereochemistry. Unless otherwise indicated, all p-amino acid abbreviations represent either isomer, i.e., the L-isomer, the D-isomer, or combinations thereof.

[0031] Gamma (y)-amino acids are amino acids with the carbon atom to which the amino group attaches is separated from the carboxylate moiety by two carbon atoms.

[0032] For additional modified and unusual amino acids, see §2422 of the MPEP, particularly Table 4 at 2400-24. Additionally, “Ac” indicates N-acetyl and “cyclo” and “c” refers to a cyclic structure. “NH2” indicates an amine group, typically added on the C- terminus of a polypeptide. Accordingly, as used herein, an — NH2 moiety on the C-terminus of a peptide indicates an amidated C-terminus.

[0033] The term “acyl” includes a group RCO — , where R is an organic group. An example is the acetyl group CH3CO — , referred to herein as “Ac.”

[0034] A peptide is “acylated” when an alkyl or substituted alkyl group as defined above is bonded through one or more carbonyl { — (C=O) — } groups. A peptide is most usually acylated at the N-terminus.

[0035] An “amine” includes compounds that contain an amine group (— NH2).-8-168785213.5

[0036] An “amide” includes compounds that have a trivalent nitrogen attached to a carbonyl group (i.e., — CO — NH2), such as for example methylamide, ethylamide, propylamide, and the like. A peptide is most usually amidated at the C-terminus by the addition of an amine ( — NH2) moiety to the C-terminal carboxyl group.

[0037] As used herein, sugar alcohols are polyols, typically derived from sugars. The term “sugar” refers to monosaccharides, disaccharides, and / or polysaccharides. Examples of sugars include, but are not limited to, sucrose, lactose, trehalose, maltose, dextrose, and the like.

[0038] The terms “active ingredient” and “active compound” refer to a biologically active substance, whether naturally or non-naturally occurring, that is the main component of the pharmaceutical composition which elicits the intended effect of an administered therapeutic. This can be any component that drives the pharmacological activity or direct effect in the diagnosis, cure, mitigation, treatment, or prevention of disease, such as cachexia.

[0039] A “formulation” or “formula” refers to the combination of substances including an active ingredient and a component, such as a pharmaceutically acceptable carrier, to form a pharmaceutical composition. The pharmaceutical composition can be formulated by any means known in the art, including but not limited to formulation as, suspensions, powders, lyophilized preparations, and may be mixed and formulated with buffers, binders, excipients, stabilizers, lubricants, oils, adjuvants, antioxidants and other agents known in the art to yield a formulation suitable for subcutaneous administration.

[0040] The formulations of the present technology are injected parenterally, such as subcutaneously. The terms “administering” or “administer” include delivery of the formulations of the present technology to a subject by parenteral administration, such as subcutaneous administration.

[0041] “Cachexia” refers to a state of general ill health and malnutrition characterized by loss of body mass including loss of weight, loss of muscle mass (skeletal, smooth, and / or cardiac muscle), loss of fat mass, or a combination thereof, and wasting. It is often associated with and induced by certain diseases or conditions such as, but not limited to,-9-168785213.5cancer, cystic fibrosis, or AIDS. The term “cancer cachexia” refers to cachexia induced by cancer.

[0042] “Anorexia” refers to a loss of appetite, whether brought on by medical, physiological, or psychological factors. Anorexia is often closely associated with, and generally contributes to, cachexia seen in patients with advanced cancers and other conditions.

[0043] As used herein, the term “isotonic” refers to fluids such as a solution having the same effective osmotic pressure as another solution, especially one in a cell or a body fluid. In some instances, an isotonic solution has the same concentration of solutes as the blood, such as an isotonic saline solution.

[0044] The terms “treat,” “treatment,” and “treating” refer to a manner of providing a pharmaceutical composition and / or melanocortin analog to alleviate disease outcomes. This includes utilizing administration techniques as described in the context of the present technology. Efficacy of treatment can be determined by measuring biomarkers as described in the context of the present technology.

[0045] The terms “subject” and “patient” refer to anyone being evaluated for disease or being administered a therapeutic or pharmaceutical composition. This includes people without diagnosed or confirmed disease. This also includes people with diagnosed or confirmed disease, such as cachexia.Pharmaceutical Compositions and Formulations

[0046] The present technology provides pharmaceutical compositions and formulations comprising a non-naturally occurring melanocortin analog and one or more pharmaceutically acceptable carriers and / or excipients.

[0047] In some embodiments, the non-naturally occurring melanocortin analog comprises the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond. In further embodiments, the non- naturally occurring melanocortin analog is in the form of a salt. In still further embodiments, the non-naturally occurring melanocortin analog is in the form of an acetate salt.-10-168785213.5

[0048] The non-naturally occurring melanocortin analog may be present in the pharmaceutical composition or formulation in an amount of about 5 mg / mL to about 200 mg / mL. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 5 mg / mL to about 150 mg / mL, about 5 mg / mL to about 130 mg / mL, about 5 mg / mL to about 100 mg / mL, about 5 mg / mL to about 70 mg / mL or about 5 mg / mL to about 50 mg / mL. In some embodiments, the non- naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 10 mg / mL to about 150 mg / mL, about 10 mg / mL to about 130 mg / mL, about 10 mg / mL to about 100 mg / mL, about 10 mg / mL to about 70 mg / mL or about 10 mg / mL to about 50 mg / mL. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 10 mg / mL to about 130 mg / mL. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 10 mg / mL to about 50 mg / mL.

[0049] In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 10 mg / mL, about 12.5 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, or about 50 mg / mL. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 12.5 mg / mL. In other embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 25 mg / mL. In still other embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in an amount of about 50 mg / mL.

[0050] Pharmaceutically acceptable carriers and / or excipients included in the pharmaceutical compositions and formulations of the present technology may be selected based on the carrier or excipient’s ability to confer desirable properties, such as reduced or lack of delivery site reactions, solubility, stability, pH, buffering, pharmacokinetics, bioavailability, and the like, to the pharmaceutical composition or formulation. For example, the one or more pharmaceutically acceptable carriers and / or excipients of the pharmaceutical compositions and formulations described herein may include a buffer, an osmolyte, and a pH adjusting agent.-11 -168785213.5

[0051] The pharmaceutical compositions and formulations of the present technology include a buffer. The buffer may be an organic buffer, i.e., a buffer that does not comprise or chelate a metal cation. In some embodiments, the buffer comprises one or more of histidine, Tris, arginine, MES, ADA, PIPES, ACES, MOPSO, MOPS, Bes, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, HEPPS, TPAS, acetoamidoglycine, glycinamide, glycylglycine, tricine, and bicine. In some embodiments, the buffer comprises histidine.

[0052] Alternatively, the buffer may be an inorganic buffer, i.e., a buffer that comprises or chelates a metal ion. In some embodiments, the buffer comprises an acetate. In further embodiments, the buffer comprises sodium acetate. A sodium acetate buffer may be included in the pharmaceutical compositions and formulations described herein as anhydrous sodium acetate or a sodium acetate hydrate.

[0053] The buffer may be present in the pharmaceutical composition or formulation at a concentration of about 5 mM to about 150 mM. In some embodiments, the buffer is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 150 mM, about 5 mM to about 125 mM, about 5 mM to about 100 mM, about 10 mM to about 120 mM, about 10 mM to about 100 mM, about 5 mM to about 80 mM, about 10 mM to about 80 mM, about 5 mM to about 50 mM, about 10 mM to about 50 mM, about 5 mM to about 25 mM, or about 10 mM to about 25 mM. In some embodiments, the buffer may be present in the pharmaceutical composition or formulation at a concentration of about 10 mM, about 12.5 mM, about 15 mM, about 17.5 mM, about 20 mM, about 22.5 mM, or about 25 mM.

[0054] In some embodiments, the buffer comprises histidine and the histidine is present in the pharmaceutical composition or formulation at a concentration of about 5 mM to about 150 mM. In some embodiments, the histidine is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 150 mM, about 5 mM to about 125 mM, about 5 mM to about 100 mM, about 10 mM to about 120 mM, about 10 mM to about 100 mM, about 5 mM to about 80 mM, about 10 mM to about 80 mM, about 5 mM to about 50 mM, about 10 mM to about 50 mM, about 5 mM to about 25 mM, or about 10 mM to about 25 mM. In some embodiments, the histidine is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 25 mM. In some embodiments, the histidine is present in the pharmaceutical composition or formulation at a-12-168785213.5concentration of about 10 mM, about 12.5 mM, about 15 mM, about 17.5 mM, about 20 mM, about 22.5 mM, or about 25 M.

[0055] In some embodiments, the buffer comprises sodium acetate and the sodium acetate is present in the pharmaceutical composition or formulation at a concentration of about 5 mM to about 150 mM. In some embodiments, the sodium acetate is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 150 mM, about 5 mM to about 125 mM, about 5 mM to about 100 mM, about 10 mM to about 120 mM, about 10 mM to about 100 mM, about 5 mM to about 80 mM, about 10 mM to about 80 mM, about 5 mM to about 50 mM, about 10 mM to about 50 mM, about 5 mM to about 25 mM, or about 10 mM to about 25 mM. In some embodiments, the sodium acetate is present in the pharmaceutical composition or formulation at a concentration of about 10 mM, about 12.5 mM, about 15 mM, about 17.5 mM, about 20 mM, about 22.5 mM, or about 25 mM.

[0056] A buffer may be included to maintain a certain pH range of the pharmaceutical composition or formulation. For example, the pharmaceutical compositions and formulations of the present technology may include a buffer to maintain a pH range that is acceptable for physiological use and that maintains dissolution of the non-naturally occurring melanocortin analog in the composition or formulation. Such pH range may be from about 4.0 to about 7.0. Accordingly, the pharmaceutical composition or formulation has a pH of about 4.0 to about 7.0. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.0 to about 6.5. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.0 to about 6.0. In some embodiments, pharmaceutical composition or formulation has a pH of about 4.0 to about 5.5. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.0 to about 5.0. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.5 to about 7.0. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.5 to about 6.5. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.5 to about 6.0. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.5 to about 5.5. In some embodiments, the pharmaceutical-13-168785213.5composition or formulation has a pH of about 5.0 to about 7.0. In some embodiments, the pharmaceutical composition or formulation has a pH of about 5.0 to about 6.5.

[0057] In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.0 to about 4.5, about 4.2 to about 4.7, about 4.4 to about 4.9, about, about 4.6 to about 5.1 , about 4.8 to about 5.3, or about 5.0 to about 5.5. In some embodiments, the pharmaceutical composition or formulation has a pH of about 5.0 to about 5.5, about 5.2 to about 5.7, about 5.4 to about 5.9, about, about 5.6 to about 6.1 , about 5.8 to about 6.3, or about 6.0 to about 6.5. In some embodiments, the pharmaceutical composition or formulation has a pH of about 4.9. In some embodiments, the pharmaceutical composition or formulation has a pH of about 5.3. In some embodiments, the pharmaceutical composition or formulation has a pH of about 6.3.

[0058] The pharmaceutical compositions and formulations of the present technology include an osmolyte. The osmolyte may be a sugar or derivative thereof, glycerol, glycerin, a gelatin, a propylene glycol, a propylparaben, PVP, PLGA, or PEG. In some embodiments, the osmolyte is a sugar or derivative thereof such as, for example, a sugar alcohol. Nonlimiting examples of sugar alcohols include sorbitol, mannitol, erythritol, threitol, lactitol, maltitol, glycerol, xylitol, and cellulose. In some embodiments, the osmolyte is mannitol.

[0059] The osmolyte may be present in the pharmaceutical composition or formulation in an amount of about 5 mg / mL to about 100 mg / mL. In some embodiments, the osmolyte is present in the pharmaceutical composition or formulation in an amount of about 10 mg / mL to about 100 mg / mL, about 10 mg / mL to about 90 mg / mL, about 10 mg / mL to about 80 mg / mL, about 10 mg / mL to about 70 mg / mL about 10 mg / mL to about 60 mg / mL, about 10 mg / mL to about 50 mg / mL, or about 10 mg / mL to about 40 mg / mL. In some embodiments, the osmolyte is present in the pharmaceutical composition or formulation in an amount of about 15 mg / mL to about 50 mg / mL, about 10 mg / mL to about 45 mg / mL, about 15 mg / mL to about 45 mg / mL, about 15 mg / mL to about 40 mg / mL about 20 mg / mL to about 50 mg / mL, about 20 mg / mL to about 45 mg / mL, or about 20 mg / mL to about 40 mg / mL.

[0060] In some embodiments, the osmolyte is present in the pharmaceutical composition or formulations in an amount of about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 27.5 mg / mL, about 30 mg / mL, about 32.5 mg / mL, about 35 -14-168785213.5mg / mL, about 37.5 mg / mL, about 40 mg / mL, about 45 mg / mL, or about 50 mg / mL. In some embodiments, the osmolyte is present in the pharmaceutical composition or formulation in an amount of about 25 mg / mL. In other embodiments, the osmolyte is present in the pharmaceutical composition or formulation in an amount of about 27.5 mg / mL. In still other embodiments, the osmolyte is present in the pharmaceutical composition or formulation in an amount of about 35 mg / mL. In yet other embodiments, the osmolyte is present in the pharmaceutical composition or formulation in an amount of about 37.5 mg / mL.

[0061] In some embodiments, the osmolyte is mannitol and the mannitol is present in the pharmaceutical composition or formulation in an amount of about 10 mg / mL to about 100 mg / mL, about 10 mg / mL to about 90 mg / mL, about 10 mg / mL to about 80 mg / mL, about 10 mg / mL to about 70 mg / mL about 10 mg / mL to about 60 mg / mL, about 10 mg / mL to about 50 mg / mL, or about 10 mg / mL to about 40 mg / mL. In some embodiments, the mannitol is present in the pharmaceutical composition or formulation in an amount of about 15 mg / mL to about 50 mg / mL, about 10 mg / mL to about 45 mg / mL, about 15 mg / mL to about 45 mg / mL, about 15 mg / mL to about 40 mg / mL about 20 mg / mL to about 50 mg / mL, about 20 mg / mL to about 45 mg / mL, or about 20 mg / mL to about 40 mg / mL.

[0062] In some embodiments, the osmolyte is mannitol and the mannitol is present in the pharmaceutical composition or formulations in an amount of about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 27.5 mg / mL, about 30 mg / mL, about 32.5 mg / mL, about 35 mg / mL, about 37.5 mg / mL, about 40 mg / mL, about 45 mg / mL, or about 50 mg / mL. In some embodiments, the mannitol is present in the pharmaceutical composition or formulation in an amount of about 25 mg / mL. In other embodiments, the osmolyte is mannitol and the mannitol is present in the pharmaceutical composition or formulation in an amount of about 27.5 mg / mL. In still other embodiments, the mannitol is present in the pharmaceutical composition or formulation in an amount of about 35 mg / mL. In yet other embodiments, the mannitol is present in the pharmaceutical composition or formulation in an amount of about 37.5 mg / mL.

[0063] An osmolyte may be included in the pharmaceutical composition or formulation to achieve a certain osmolality. For example, the pharmaceutical compositions and formulations of the present technology may include an osmolyte to achieve an isotonic or-15-168785213.5hypertonic solution to minimize injection site pain and cellular damage. An isotonic solution may have an osmolality of about 250 mOsm / kg to about 350 mOsm / kg and a hypertonic solution may have an osmolality of about 350 mOsm / kg to about 600 mOsm / kg. Accordingly, in some embodiments, the osmolyte is included in the pharmaceutical composition or formulation to achieve an osmolality of about 250 mOsm / kg to about 600 mOsm / kg. In some embodiments, the osmolyte is included in the pharmaceutical composition or formulation to achieve an osmolality of about 280 mOsm / kg to about 560 m / Osm, about 280 mOsm / kg to about 520 m / Osm, about 280 mOsm / kg to about 500 mOsm, about 280 mOsm / kg to about 460 m / Osm, about 280 mOsm / kg to about 420 m / Osm, about 280 mOsm / kg to about 400 mOsm, about 280 mOsm / kg to about 360 m / Osm, or about 280 mOsm / kg to about 320 m / Osm.

[0064] In some embodiments, the pharmaceutical composition or formulation is isotonic. In further embodiments, the osmolality of the pharmaceutical composition of formulation is about 250 mOsm / kg to about 350 mOsm / kg. In some embodiments, the osmolality of the pharmaceutical composition or formulation is about 260 mOsm / kg to about 330 mOsm / kg, 270 mOsm / kg to about 330 mOsm / kg, about 275 mOsm / kg to about 310 mOsm / kg, about 275 mOsm / kg to about 300 mOsm / kg, or about 280 mOsm / kg to about 290 mOsm / kg. In some embodiments, the osmolality of the pharmaceutical composition or formulation is about 280 mOsm / kg, 281 mOsm / kg, 282 mOsm / kg, 283 mOsm / kg, 284 mOsm / kg, 286 mOsm / kg, 287 mOsm / kg, 288 mOsm / kg, 289 mOsm / kg, or 290 mOsm / kg.

[0065] In some embodiments, the pharmaceutical composition or formulation is hypertonic. In further embodiments, the osmolality of the pharmaceutical composition of formulation is about 350 mOsm / kg to about 600 mOsm / kg. In some embodiments, the osmolality of the pharmaceutical composition or formulation is about 360 mOsm / kg to about 550 mOsm / kg, about 365 mOsm / kg to about 530 mOsm / kg, about 370 mOsm / kg to about 510 mOsm / kg, about 375 mOsm / kg to about 500 mOsm / kg, about 380 mOsm / kg to about 490 mOsm / kg, about 385 mOsm / kg to about 470 mOsm / kg, or about 390 mOsm / kg to about 450 mOsm / kg. In some embodiments, the osmolality of the pharmaceutical composition or formulation is about 400 mOsm / kg to about 600 mOsm / kg, about 420 mOsm / kg to about-16-168785213.5580 mOsm / kg, about 440 mOsm / kg to about 560 mOsm / kg, about 460 mOsm / kg to about 540 mOsm / kg, or about 480 mOsm / kg to about 520 mOsm / kg.

[0066] The pharmaceutical compositions and formulations of the present technology include a pH adjusting agent. The pH adjusting agent may be an acid or a base. For example, if the pH of the pharmaceutical composition or formulation is too basic before addition of the pH adjusting agent, then the pH adjusting agent may be an acid. On the other hand, if the pH of the pharmaceutical composition or formulation is too acidic before addition of the pH adjusting agent, then the pH adjusting agent may be a base.

[0067] The pH adjusting agent may be present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 10 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 0.1 mg / mL to about 9.0 mg / mL, about 0.1 mg / mL to about 8.0 mg / mL, about 0.1 mg / mL to about 7.0 mg / mL, about 0.1 mg / mL to about 6.0 mg / mL, about 0.1 mg / mL to about 5.0 mg / mL, about 0.1 mg / mL to about 4.0 mg / mL, about 0.1 mg / mL to about 3.0 mg / mL, or about 0.1 mg / mL to about 2.0 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 10 mg / mL, about 0.2 mg / mL to about 10 mg / mL, about 1.0 mg / mL to about 5.0 mg / mL, 0.5 mg / mL to about 5.0 mg / mL, or about 02. mg / mL to about 5 mg / mL.

[0068] When the non-naturally occurring melanocortin analog is in the form of an acetate salt, the ratio of acetate to analog may be 1 :1. Alternatively, the ratio of acetate to analog may not be 1 :1 , i.e., the ratio of acetate to analog may be less than or greater than 1 :1 . Accordingly, when a melanocortin analog acetate salt has a high ratio of acetate to analog, the pharmaceutical composition may be more acidic and may require a greater amount of the pH adjusting agent. Alternatively, when a melanocortin analog acetate salt has a low ratio of acetate to analog, the pharmaceutical composition may be less acidic and may require a lower amount of the pH adjusting agent.

[0069] In some embodiments, the pH adjusting agent is present in the pharmaceutical composition in an amount of about 0.01 mg / mL to about 5.0 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an -17-168785213.5amount of about 0.01 mg / mL to about 4.0 mg / mL, about 0.01 mg / mL to about 3.0 mg / mL, about 0.01 mg / mL to about 2.0 mg / mL, about 0.1 mg / mL to about 5.0 mg / mL, about 0.1 mg / mL to about 4.0 mg / mL, about 0.1 mg / mL to about 3.0 mg / mL, or about 0.1 mg / mL to about 2.0 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition in an amount of about 0.1 mg / mL to about 2.0 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 0.1 mg / mL to about 1 .8 mg / mL, about 0.1 mg / mL to about 1.6 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.2 mg / mL to about 1.8 mg / mL, about 0.2 mg / mL to about 1.6 mg / mL, or about 0.2 mg / mL to about 1.5 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL to about 1 .4 mg / mL, about 0.4 mg / mL to about 1.2 mg / mL, about 0.4 mg / mL to about 1.0 mg / mL, about 0.4 mg / mL to about 0.8 mg / mL, about 0.4 mg / mL to about 0.6 mg / mL, about 0.8 mg / mL to about 1 .4 mg / mL about 0.9 mg / mL to about 1 .3 mg / mL, or about 1 .0 mg / mL to about 1 .2 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1 .0 mg / mL, about 1 .1 mg / mL, about 1 .2 mg / mL, or about 1 .4 mg / mL.

[0070] In some embodiments, the pH adjusting agent is present in the pharmaceutical composition in an amount of about 1.0 mg / mL to about 5.0 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 4.5 mg / mL, about 1 .5 mg / mL to about 4.0 mg / mL, about 2.0 mg / mL to about 3.5 mg / mL, or about 2.5 mg / mL to about 3.0 mg / mL. In some embodiments, the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 1 .5 mg / mL, about 1 .0 mg / mL to about 3.0 mg / mL, about 1 .0 mg / mL to about 5.0 mg / mL about 1 .0 mg / mL to about 2.5 mg / mL, or about 2.5 mg / mL to about 5.0 mg / mL.

[0071] In some embodiments, the pH adjusting agent is a base. Non-limiting examples of bases include histidine, arginine, sodium hydroxide, potassium hydroxide, ammonium-18-168785213.5hydroxide, ammonium carbonate, sodium bicarbonate, sodium carbonate, and sodium borate. In some embodiments, the base is sodium hydroxide.

[0072] In some embodiments, the pH adjusting agent is sodium hydroxide and the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 10 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 9.0 mg / mL, about 0.01 mg / mL to about 8.0 mg / mL, about 0.01 mg / mL to about 7.0 mg / mL, about 0.01 mg / mL to about 6.0 mg / mL, about 0.01 mg / mL to about 5.0 mg / mL, about 0.01 mg / mL to about 4.0 mg / mL, about 0.01 mg / mL to about 3.0 mg / mL, or about 0.01 mg / mL to about 2.0 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.1 mg / mL to about 9.0 mg / mL, about 0.1 mg / mL to about 8.0 mg / mL, about 0.1 mg / mL to about 7.0 mg / mL, about 0.1 mg / mL to about 6.0 mg / mL, about 0.1 mg / mL to about 5.0 mg / mL, about 0.1 mg / mL to about 4.0 mg / mL, about 0.1 mg / mL to about 3.0 mg / mL, or about 0.1 mg / mL to about 2.0 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 10 mg / mL, about 0.2 mg / mL to about 10 mg / mL, about 1 .0 mg / mL to about 5.0 mg / mL, 0.5 mg / mL to about 5.0 mg / mL, or about 0.2 mg / mL to about 5 mg / mL.

[0073] In some embodiments, the sodium hydroxide is present in an amount of about 0.1 mg / mL to about 2.0 mg / mL, 0.1 mg / mL to about 1 .8 mg / mL, about 0.1 mg / mL to about 1.6 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.2 mg / mL to about 1.8 mg / mL, about 0.2 mg / mL to about 1.6 mg / mL, or about 0.2 mg / mL to about 1.5 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL to about 1 .4 mg / mL, about 0.4 mg / mL to about 1.2 mg / mL, about 0.4 mg / mL to about 1.0 mg / mL, about 0.4 mg / mL to about 0.8 mg / mL, about 0.4 mg / mL to about 0.6 mg / mL, about 0.8 mg / mL to about 1 .4 mg / mL about 0.9 mg / mL to about 1.3 mg / mL, or about 1.0 mg / mL to about 1.2 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1 .0 mg / mL, about 1 .1 mg / mL, about 1 .2 mg / mL,-19-168785213.5or about 1 .4 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.4 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.5 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .2 mg / mL.

[0074] In some embodiments, the sodium hydroxide is present in the pharmaceutical composition in an amount of about 1.0 mg / mL to about 5.0 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 4.5 mg / mL, about 1 .5 mg / mL to about 4.0 mg / mL, about 2.0 mg / mL to about 3.5 mg / mL, or about 2.5 mg / mL to about 3.0 mg / mL. In some embodiments, the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 1 .5 mg / mL, about 1 .0 mg / mL to about 3.0 mg / mL, about 1 .0 mg / mL to about 5.0 mg / mL about 1 .0 mg / mL to about 2.5 mg / mL, or about 2.5 mg / mL to about 5.0 mg / mL.

[0075] In some embodiments, the pH adjusting agent is histidine and the histidine is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 10 mg / mL. In some embodiments, the histidine is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / mL, about 0.01 mg / mL to about 4.0 mg / mL, about 0.01 mg / mL to about 3.0 mg / mL, about 0.01 mg / mL to about 2.0 mg / mL, or about 0.01 mg / mL to about 1.0 mg / mL. In some embodiments, the histidine is present in the pharmaceutical composition or formulation in an amount of about 0.1 mg / mL to about 5.0 mg / mL, about 0.1 mg / mL to about 4.0 mg / mL, about 0.1 mg / mL to about 3.0 mg / mL, about 0.1 mg / mL to about 2.0 mg / mL, about 0.5 mg / mL to about 5.0 mg / mL, about 0.5 mg / mL to about 4.0 mg / mL, about 0.5 mg / mL to about 3.0 mg / mL, or about 0.5 mg / mL to about 2.0 mg / mL. In some embodiments, the histidine is present in the pharmaceutical composition or formulation in an amount of about 1.0 mg / mL to about 5.0 mg / mL, about 1.0 mg / mL to about 4.0 mg / mL, about 1.0 mg / mL to about 3.0 mg / mL, or about 1 .0 mg / mL to about 2.0 mg / mL. .-20-168785213.5

[0076] In some embodiments, the histidine is present in an amount of about 0.1 mg / mL to about 2.0 mg / mL, 0.1 mg / mL to about 1 .8 mg / mL, about 0.1 mg / mL to about 1 .6 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.2 mg / mL to about 1.8 mg / mL, about 0.2 mg / mL to about 1 .6 mg / mL, or about 0.2 mg / mL to about 1 .5 mg / mL. In some embodiments, the histidine is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL to about 1.4 mg / mL, about 0.4 mg / mL to about 1.2 mg / mL, about 0.4 mg / mL to about 1 .0 mg / mL, about 0.4 mg / mL to about 0.8 mg / mL, about 0.4 mg / mL to about 0.6 mg / mL, about 0.8 mg / mL to about 1 .4 mg / mL about 0.9 mg / mL to about 1 .3 mg / mL, or about 1 .0 mg / mL to about 1 .2 mg / mL. In some embodiments, the histidine is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1 .0 mg / mL, about 1 .1 mg / mL, about 1 .2 mg / mL, or about 1 .4 mg / mL.

[0077] In some embodiments, the pH adjusting agent is an acid. Non-limiting examples of acids include acetic acid, citric acid, fumaric acid, hydrochloric acid, and nitric acid. In some embodiments, the acid is acetic acid, such as glacial acetic acid.

[0078] In some embodiments, the pH adjusting agent is glacial acetic acid and the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / mL, about 0.01 mg / mL to about 4.0 mg / mL, about 0.01 mg / mL to about 3.0 mg / mL, about 0.01 mg / mL to about 2.0 mg / mL, or about 0.01 mg / mL to about 1 .0 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.1 mg / mL to about 9.0 mg / mL, about 0.1 mg / mL to about 8.0 mg / mL, about 0.1 mg / mL to about 7.0 mg / mL, about 0.1 mg / mL to about 6.0 mg / mL, about 0.1 mg / mL to about 5.0 mg / mL, about 0.1 mg / mL to about 4.0 mg / mL, about 0.1 mg / mL to about 3.0 mg / mL, or about 0.1 mg / mL to about 2.0 mg / mL.. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 10 mg / mL, about 0.2 mg / mL to about 10 mg / mL, about 1 .0 mg / mL to about 5.0 mg / mL, 0.5 mg / mL to about 5.0 mg / mL, or about 0.2 mg / mL to about 5 mg / mL..-21 -168785213.5

[0079] In some embodiments, the glacial acetic acid is present in an amount of about 0.1 mg / mL to about 2.0 mg / mL, 0.1 mg / mL to about 1 .8 mg / mL, about 0.1 mg / mL to about 1.6 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.2 mg / mL to about 1.8 mg / mL, about 0.2 mg / mL to about 1.6 mg / mL, or about 0.2 mg / mL to about 1.5 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL to about 1 .4 mg / mL, about 0.4 mg / mL to about 1.2 mg / mL, about 0.4 mg / mL to about 1.0 mg / mL, about 0.4 mg / mL to about 0.8 mg / mL, about 0.4 mg / mL to about 0.6 mg / mL, about 0.8 mg / mL to about 1 .4 mg / mL about 0.9 mg / mL to about 1.3 mg / mL, or about 1.0 mg / mL to about 1.2 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.2 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1 .0 mg / mL, about 1 .1 mg / mL, about 1 .2 mg / mL, or about 1.4 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.4 mg / mL, including about 0.42 mg / mL to about 0.44 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 1 .1 mg / mL, including about 1.13 mg / mL to about 1 .15 mg / mL.

[0080] In some embodiments, the glacial acetic acid is present in the pharmaceutical composition in an amount of about 1.0 mg / mL to about 5.0 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 4.5 mg / mL, about 1 .5 mg / mL to about 4.0 mg / mL, about 2.0 mg / mL to about 3.5 mg / mL, or about 2.5 mg / mL to about 3.0 mg / mL. In some embodiments, the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 1 .0 mg / mL to about 1 .5 mg / mL, about 1 .0 mg / mL to about 3.0 mg / mL, about 1 .0 mg / mL to about 5.0 mg / mL about 1 .0 mg / mL to about 2.5 mg / mL, or about 2.5 mg / mL to about 5.0 mg / mL.

[0081] The pharmaceutical compositions and formulations of the present technology further comprise water. The water may be sterile water for injection (SWFI), sterile water, bacteriostatic water for injection (BWFI), distilled water, bidistilled water, deionized water,-22-168785213.5deionized distilled water, and reverse osmosis water. In some embodiments, the water present in the pharmaceutical composition or formulation is sterile water for injection.

[0082] The pharmaceutical compositions and formulations of the present technology may further comprise a surfactant. Non-limiting examples of surfactant that may be included in the pharmaceutical compositions or formulations include anionic surfactants such as alkyl carboxylate-fatty acid salts, sulfates (e.g., sodium lauryl sulfate and alkyl ether sulfates), sulfonates (e.g., dioctyl sodium sulfosuccinate and alkyl benzenesulfonates), and phosphate esters (e.g., alkyl aryl ether phosphates and alkyl ether phosphates); cationic surfactants such as quaternary ammoniums (e.g., cetrimonium bromide, cetylpyridinium chloride, and dimethyldioctadecylammonium chloride), amine-based surfactants (e.g., triethylamine hydrochloride and octenidine dihydrochloride) and pyridinium surfactants (e.g., benzethonium chloride); non-ionic surfactants such as polyol esters (e.g., fatty acid esters of sorbitan), polyoxyethylene esters (e.g., polysorbates), and poloxamers (e.g., poloxamer 188); and amphoteric surfactants such as phospholipids (e.g., phosphatidylcholine and lecithin), carboxylic acid / quaternary ammoniums (e.g., cocamidopropyl betaine and amidosulfobetaine-16), phosphoric acid / quaternary ammoniums (e.g., hexadecyl phosphocholine), and betaines (e.g., alkylamidopropyl betaine). In some embodiments, the surfactant is a non-ionic surfactant. In further embodiments, the surfactant is a polysorbate.

[0083] Additionally, or alternatively, the pharmaceutical compositions and formulations of the present technology may further comprise a preservative. Non-limiting examples of preservatives that may be included in the pharmaceutical compositions or formulations include m-cresol, phenol, benzyl alcohol, chlorobutanol, methylparaben, phenoxyethanol, propylparaben, and thimerosal. In some embodiments, the preservative is selected from m- cresol, phenol, and benzyl alcohol. In some embodiments, the preservative is m-cresol. In some embodiments, the preservative is phenol. In some embodiments, the preservative is benzyl alcohol.

[0084] A preservative may be included in the pharmaceutical compositions and formulations described herein in an amount sufficient to maintain stability of the composition or formulation over time. For example, the preservative may be present in the pharmaceutical compositions and formulations in an amount of about 0.1 wt% to about 1.0-23-168785213.5wt%, relative to a total weight of the composition or formulation. In some embodiments, the preservative is present in the pharmaceutical compositions and formulations in an amount of about 0.1 wt% to about 0.8 wt%, about 0.2 wt% to about 0.6 wt%, or about 0.3 wt% to about 0.5 wt%, relative to a total weight of the composition or formulation. In some embodiments, the preservative is present in the pharmaceutical compositions and formulations in an amount of about 0.3 wt%, relative to a total weight of the composition or formulation. In some embodiments, the preservative is present in the pharmaceutical compositions and formulations in an amount of about 0.5 wt%, relative to a total weight of the composition or formulation.

[0085] In some embodiments, the pharmaceutical composition or formulation comprises substantially no chloride ions. In some embodiments, the pharmaceutical composition or formulation is free from chloride ions.

[0086] In some embodiments, the pharmaceutical composition or formulation comprises substantially no metal ions. In some embodiments, the pharmaceutical composition or formulation is free from metal ions.

[0087] In some embodiments, the pharmaceutical composition has a pH of about 5.3, and comprises a non-naturally occurring melanocortin analog in an amount of 12.5 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]- dVal-dPro-NH2 (SEQ ID NO: 2), mannitol in an amount of about 30 mg / mL to about 40 mg / mL, an acid in an amount of about 1.3 mg / mL to about 1.5 mg / mL, and histidine at a concentration of about 25 mM, wherein c in the sequence represents cyclization via a lactam bond. In further embodiments, the mannitol is present in the composition in an amount of about 37.5 mg / mL. In still further embodiments, the acid is glacial acetic acid.

[0088] In some embodiments, the pharmaceutical composition has a pH of about 5.3, and comprises a non-naturally occurring melanocortin analog in an amount of 25 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]- dVal-dPro-NH2 (SEQ ID NO: 2), mannitol in an amount of about 30 mg / mL to about 40 mg / mL, an acid in an amount of about 0.42 mg / mL to about 0.44 mg / mL, and histidine at a concentration of about 25 mM, wherein c in the sequence represents cyclization via a lactam-24-168785213.5bond. In further embodiments, the mannitol is present in the composition in an amount of about 35 mg / mL. In still further embodiments, the acid is glacial acetic acid.

[0089] In some embodiments, the pharmaceutical composition has a pH of about 4.0 to about 6.5, and comprises a non-naturally occurring melanocortin analog in an amount of 50 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg- Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), mannitol in an amount of about 25 mg / mL to about 35 mg / mL, a base in an amount of about 0.4 mg / mL to about 1 .2 mg / mL, and a buffer at a concentration of about 25 mM, wherein c in the sequence represents cyclization via a lactam bond. In some embodiments, the mannitol is present in the composition in an amount of about 25 mg / mL. In other embodiments, the mannitol is present in the composition in an amount of about 27.5 mg / mL. In further embodiments, the base is sodium hydroxide. In some embodiments, the buffer is histidine. In other embodiments, the buffer is sodium acetate. In some embodiments, the sodium hydroxide is present in an amount of 0.4 mg / mL, the buffer is histidine, and the pH of the pharmaceutical composition is about 5.3. In other embodiments, the sodium hydroxide is present in an amount of 1 .2 mg / mL, the buffer is histidine, and the pH of the pharmaceutical composition is about 6.3. In still other embodiments, the sodium hydroxide is present in an amount of 0.5 mg / mL, the buffer is sodium acetate, and the pH of the pharmaceutical composition is about 5.0.Methods of Preparing Pharmaceutical Compositions and Formulations

[0090] The present technology further provides methods of preparing pharmaceutical compositions and formulations described herein, e.g., pharmaceutical compositions comprising a non-naturally occurring melanocortin analog and one or more pharmaceutically acceptable carriers and / or excipients. In some embodiments, the method produces a pharmaceutical composition or formulation of a non-naturally occurring melanocortin analog that has a pH of about 4.0 to about 6.5.

[0091] Methods of preparing the pharmaceutical compositions and formulations comprise providing an aqueous peptide composition of the non-naturally occurring melanocortin analog. In some embodiments, the aqueous peptide composition is provided by dissolving the non-naturally occurring melanocortin analog in water. In some embodiments, when a surfactant is included in the pharmaceutical composition or -25-168785213.5formulation, the surfactant is also dissolved in the water to provide the aqueous peptide composition. A surfactant may be included in the aqueous peptide composition to prevent the peptide from adhering to the walls of the reaction vessel or tubing.

[0092] In some embodiments, the non-naturally occurring melanocortin analog has the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond. In further embodiments, the non-naturally occurring melanocortin analog is in the form of a salt. In still further embodiments, the non- naturally occurring melanocortin analog is in the form of an acetate salt. To achieve complete dissolution of the non-naturally occurring melanocortin analog in the water, the aqueous peptide solution may be stirred for a period of time sufficient to achieve a homogenous solution. The period of time sufficient to achieve homogeneity may depend on the batch size of the pharmaceutical composition or formulation. In some embodiments, the aqueous peptide composition is stirred for about 10 minutes to about 24 hours. In some embodiments, the aqueous peptide composition is stirred for about 10 minutes to about 60 minutes, about 15 minutes to about 50 minutes, about 20 minutes to about 40 minutes. In some embodiments, the aqueous peptide composition is stirred for about 30 minutes to achieve homogeneity of the composition. Alternatively, in some embodiments, the aqueous peptide composition is stirred for about 1 hour to about 24 hours. In some embodiments, the aqueous peptide composition is stirred for about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, or about 24 hours to achieve homogeneity of the composition. The aqueous peptide composition may be mixed and maintained at ambient temperature.

[0093] When the non-naturally occurring melanocortin analog is in the form of a salt, such as, for example, an acetate salt, the aqueous peptide composition may be acidic. In some embodiments, the aqueous peptide composition has a pH of about 2.5 to about 5.0. In some embodiments, the aqueous peptide composition has a pH of about 2.5, about 2.8, about 3.0, about 3.2, about 3.5, about 3.8, about 4.0, about 4.2, about 4.5, about 4.8, or about 5.0.-26-168785213.5

[0094] Methods of preparing the pharmaceutical compositions and formulations comprise providing a pharmaceutically acceptable carrier composition. In some embodiments, the pharmaceutically acceptable carrier composition is provided by dissolving one or more pharmaceutically acceptable carriers and / or excipients in water. In further embodiments, the pharmaceutically acceptable carrier composition is provided by dissolving an osmolyte, a pH adjusting agent, and a buffer in water. In some embodiments, when a surfactant is included in the pharmaceutical composition or formulation, the surfactant is also dissolved in the water to provide the pharmaceutically acceptable carrier composition. The pharmaceutically acceptable carrier composition may be mixed for an amount of time sufficient to achieve a homogenous composition. The period of time sufficient to achieve homogeneity may depend on the batch size of the pharmaceutical composition or formulation. In some embodiments, the aqueous peptide composition is stirred for about 5 minutes to about 24 hours. In some embodiments, the aqueous peptide composition is stirred for about 5 minutes to about 40 minutes, about 10 minutes to about 30 minutes, about 15 minutes to about 20 minutes. In some embodiments, the aqueous peptide composition is stirred for about 15 minutes to achieve homogeneity of the composition. Alternatively, in some embodiments, the aqueous peptide composition is stirred for about 1 hour to about 24 hours. In some embodiments, the aqueous peptide composition is stirred for about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, or about 24 hours to achieve homogeneity of the composition. The pharmaceutically acceptable carrier composition may be mixed and maintained at ambient temperature.

[0095] The pharmaceutically acceptable carrier composition may have a pH that is sufficiently basic to neutralize the aqueous peptide composition, e.g., a pH that is sufficiently basic to produce a pharmaceutical composition or formulation having a pH of about 4.0 to about 7.0. For example, the pharmaceutically acceptable carrier composition may have a pH of about 5.0 to about 11 .0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 7.0 to about 1 1 .0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 8.0 to about 11 .0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 9.0 to about 11.0. In some embodiments, the pharmaceutically acceptable carrier -27-168785213.5composition has a pH of about 10.0 to about 11.0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 10.5 to about 11.0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 8.0 to about 10.5. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 8.5 to about 10.0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 9.0 to about 9.5.

[0096] In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 4.0 to about 7.0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 4.0 to about 6.5. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 4.5 to about 6.5. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 5.5 to about 6.5. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 4.5 to about 6.0. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 4.5 to about 5.5. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 5.5 to about 5.9. In some embodiments, the pharmaceutically acceptable carrier composition has a pH of about 6.2 to about 6.6.

[0097] Methods of preparing the pharmaceutical compositions and formulations comprise combining an aqueous peptide composition with a pharmaceutically acceptable carrier composition. In some embodiments, the aqueous peptide composition is added to the pharmaceutically acceptable carrier composition. In other embodiments, the pharmaceutically acceptable carrier composition is added to the aqueous peptide composition. Once the aqueous peptide composition and the pharmaceutically acceptable carrier composition are combined, the combined solution may be stirred for an amount of time sufficient to achieve a stable homogeneous composition. In some embodiments, the combined solution is stirred for about 10 minutes to about 24 hours. In some embodiments, the aqueous peptide composition is stirred for about 10 minutes to about 60 minutes, about 15 minutes to about 50 minutes, about 20 minutes to about 40 minutes. In some embodiments, the combined solution is stirred for about 30 minutes to achieve homogeneity. Alternatively, in some embodiments, the aqueous peptide composition is stirred for about 1-28-168785213.5hour to about 24 hours. In some embodiments, the aqueous peptide composition is stirred for about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, or about 24 hours to achieve homogeneity of the composition. The combined solution may be stirred at ambient temperature.

[0098] While the aqueous peptide composition and the pharmaceutically acceptable carrier composition are being combined, a local pH of the pharmaceutical composition may be maintained such that no local region has a pH that is equal to the isoelectric point of the melanocortin analog so as to prevent aggregation of the melanocortin analog. As used herein, a “local pH” refers to the pH of the composition surrounding the site where the composition is being formed, i.e., where the aqueous peptide composition and the pharmaceutically acceptable carrier composition are combined. The area of the site where the local pH is maintained may be relative to the process used to combine the aqueous peptide composition and the pharmaceutically acceptable carrier composition. For example, if the pharmaceutically acceptable carrier composition is poured into the aqueous peptide composition, or vice versa, the area of the site where the local pH is maintained may be larger than the area of the site where the local pH is maintained if the pharmaceutically acceptable carrier composition is cannulated, syringed, or pipetted into the aqueous peptide composition, or vice versa. Methods described herein may prevent aggregation of the non- naturally occurring melanocortin analog by combining the aqueous peptide composition with the pharmaceutically acceptable carrier composition via slow addition, such as for example, drop-wise addition, timed addition, portioned addition, and the like.

[0099] In some embodiments, once the aqueous peptide composition and the pharmaceutically acceptable carrier composition are combined and a homogenous solution is provided, the pH of the solution is adjusted to achieve the desired pH of the pharmaceutical composition or formulation. In further embodiments, the desired pH of the pharmaceutical composition or formulation is about 4.0 to about 6.5. In other embodiments, the pH of the homogenous solution is the desired pH of the pharmaceutical composition or formulation. Accordingly, in some embodiments, the pH is not adjusted.-29-168785213.5

[0100] In some embodiments, the pharmaceutical composition or formulations produced by the methods described herein comprise substantially no precipitated non- naturally occurring melanocortin analog. In some embodiments, the pharmaceutical composition or formulations produced by the methods described herein are free from precipitated non-naturally occurring melanocortin analog.

[0101] In some embodiments, the method is a method of producing a pharmaceutical composition of a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, the method comprising combining an aqueous peptide composition of the non-naturally occurring melanocortin analog with a pharmaceutically acceptable carrier composition having a pH of about 5.5 to about 11 .0 and comprising an osmolyte, a pH adjusting agent, and a buffer, wherein the pharmaceutical composition has a pH of about 4.0 to about 6.5.

[0102] In some embodiments, the method is a method of producing a pharmaceutical formulation having a pH of about 4.0 to about 6.5, comprising combining an aqueous peptide composition of a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, with a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer; and preventing aggregation of the analog by maintaining a local pH of the pharmaceutical composition such that no local region has a pH that is equal to the isoelectric point of the analog while the aqueous peptide composition and the pharmaceutically acceptable carrier composition are being combined.

[0103] In some embodiments, the method is a method of producing a pharmaceutical formulation of a non-naturally occurring melanocortin analog having a pH of about 4.0 to about 6.5, comprising combining an aqueous peptide composition of the non-naturally occurring melanocortin analog with a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer, wherein the non-naturally occurring melanocortin analog has the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]- dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond and the pharmaceutically acceptable carrier composition has a pH of about 5.5 to about 11 .0.-30-168785213.5EXAMPLES

[0104] The following examples are intended to illustrate various embodiments of the present technology. As such, the specific embodiments discussed are not to be construed as limitations on the scope of the present technology. It will be apparent to one skilled in the art that various equivalents, changes, and modifications may be made without departing from the scope of present technology, and it is understood that such equivalent embodiments, are to be included herein. Further, all references cited herein are hereby incorporated by reference in their entirety, as if fully set forth herein.Example 1: Peptide Synthesis-Generic

[0105] The non-naturally occurring melanocortin analogs described herein were synthesized by conventional procedures (e.g., solution-phase procedure, solid-phase synthesis) for the formation of a peptide linkage between amino acids. The solution-phase procedure involved a condensation between the free alpha amino group of an amino acid or derivative thereof having the carboxyl group or other reactive groups protected and the free primary carboxyl group of another amino acid or derivative thereof having the amino group or other reactive groups protected. The solid-phase synthesis utilized a variety of resins and reagents and may involve additional purification steps.

[0106] The process for synthesizing the non-naturally occurring melanocortin analogs was generally performed by a procedure as follows. Each amino acid in the desired sequence of the non-naturally occurring melanocortin analogs was added one at a time in succession to another amino acid or derivative thereof or by a procedure whereby peptide fragments with the desired amino acid sequence were first synthesized conventionally and then condensed to provide the desired peptide. In most cases, the resulting peptide was then cyclized to yield a cyclic peptide.

[0107] Solid-phase peptide synthesis was carried out by sequentially incorporating the desired amino acid residues one at a time into the growing peptide chain coupled to a solidphase support according to the general principles of solid phase methods (see Merrifield, Angew Chem. 24:799-810 (1985) and Barany et al., The Peptides, Analysis, Synthesis and Biology, Vol. 2, Gross E. and Meienhofer J., Eds. Academic Press 1 -284-31 -168785213.5(1980)). An exemplary solid-phase synthesis of non-naturally occurring melanocortin analogs is provided below.

[0108] Initially, the C-terminal amino acid residue of the non-naturally occurring melanocortin analog was coupled to a support, e.g., a solid-phase or tag support. Coupling of the C-terminal amino acid residue and the solid-phase support may be carried out according to any method know in the art. Depending on the coupling method, the alphaamine of the C-terminal amino acid residue may or may not be protected with an amine protecting group, as described below. Likewise, the carboxyl group of the amino acid residue may or may not be activated prior to coupling to the solid-phase support in order to increase its electrophilicity. Some methods of coupling rely on the formation of an ester bond between the carboxyl group of the amino acid and a reactive handle on the solid-phase resin. For example, an amino acid residue may be coupled to a p-benzyloxybenzyl alcohol resin (Wang) or a 2-chlorotrityl chloride resin via an ester linkage. Some methods of coupling rely on the formation of an amide bond between the carboxyl group of the amino acid and a reactive handle on the solid-phase resin For example, an amino acid residue may be coupled to a benzhydrylamine (BHA) resin through an Fmoc-linker such as, for example, p- [(R,S)-a-[1 -(9H-fluor-en-9-yl)-methoxyformamido]-2,4-dimethyloxybenzyl]-phenoxyacetic acid (Rink linker) via an amide linkage.

[0109] The non-naturally occurring melanocortin analog was then synthesized by sequential amino acid addition or combination of peptide fragments. Subsequently, the peptide was cleaved from the solid-phase support and purified by methods known in the art, such as, for example, reverse phase high performance liquid chromatography (RP-HPLC) using a suitable column, such as a C18 column, or crystallization. Additionally, or alternatively, other methods of separation or purification were employed, including, but not limited to, methods based on the size or charge of the peptide, and affinity tag-based purification. Once purified, the peptide was characterized by methods such as high- performance liquid chromatograph (HPLC), amino acid analysis, mass spectrometry, and the like.Example 2: Peptide Synthesis-Protecting Groups-32-168785213.5

[0110] During synthesis of the non-naturally occurring melanocortin analogs, reactive side chain groups of the various amino acid residues were protected with suitable protecting groups, which prevented undesirable chemical reaction from occurring at that site until the protecting group was removed.

[0111] Additionally, protection of the alpha amino group of an amino acid residue or fragment was performed while that entity reacting with the carboxyl group, followed by the selective removal of the alpha amino protecting group to allow a subsequent reaction to take place at that site. Specific protecting groups for solid phase synthesis methods and solution phase synthesis methods are known to those having ordinary skill in the art. Alpha amino groups were protected by a suitable protecting group, including a urethane-type protecting group, such as benzyloxycarbonyl (Z) and substituted benzyloxycarbonyl, such as p- chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p- biphenyl-isopropoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc) and p- methoxybenzyloxycarbonyl (Moz); aliphatic urethane-type protecting groups, such as t- butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropoxycarbonyl, and allyloxycarbonyl. Fmoc was also used for alpha amino protection. Guanidino groups, if present, were protected by a suitable protecting group, such as nitro, p-toluenesulfonyl (Tos), Z, pentamethylchromanesulfonyl (Pmc), adamantyloxycarbonyl, pentamethyldihydrobenzofuran-5-sulfonyl (Pbf) and Boc. Pmc was used as a protecting group for Arg.

[0112] Alpha amino protecting groups may be removed under basic conditions, such as, for example, using a solution of piperidine, piperazine, diethylamine, or morpholine (20- 40% v / v) in N,N-dimethylformamide (DMF). In synthesis methods in which alpha amino protecting groups were used, protecting groups were removed after synthesis of the peptide and before or after cleavage of the solid-phase support.Example 3: Peptide Synthesis-Additional Modifications

[0113] If necessary, the peptides were further modified to obtain N-terminus modifications, such as acetylation, while on resin, or were removed from the resin by use of a cleaving reagent and then modified. Likewise, C-terminus modification (e.g., amidation), was performed if needed.-33-168785213.5

[0114] Additionally, the cyclized peptide structures were obtained prior to cleavage from the peptide resin. For cyclization through reactive side chain moieties, the desired side chains were deprotected, and the peptide suspended in a suitable solvent and a cyclic coupling agent added. Suitable solvents, for example DMF, dichloromethane (DCM) or 1 - methyl-2-pyrrolidone (NMP), were used for the cyclization. Suitable cyclic coupling reagents (e.g., 2-(1 H-benzotriazol-1 -yl)-1 ,1 ,3,3-tetramethyluronium tetrafluoroborate (TBTLI), 2-(1 H- benzotriazol-1 -yl)-1 ,1 ,3,3-tetramethyluronium hexafluorophosphate (HBTLI), benzotriazole- 1 -yl-oxy-tris(dimethylamino)phosphoniumhexafluorophosphate (BOP), benzotriazole-1 -yl- oxy-tris(pyrrolidino)phosphoniumhexafluorophosphate (PyBOP), 2-(7-aza-1 H-benzotriazol- 1 -yl)-1 ,1 ,3,3-tetramethyluronium tetrafluoroborate (TATU), 2-(2-oxo-1 (2H)-pyridyl)-1 ,1 ,3,3- tetramethyluronium tetrafluoroborate (TPTLI), N,N'-dicyclohexylcarbodiimide / 1 - hydroxybenzotriazole (DCCI / HOBt) hexafluorophosphate azabenzotriazole tetramethyl uranium (HATU), or ethyl (hydroxyimino)cyanoacetate / diisopropylcarbodiimide (Oxyma / DIG)) were also used for the cyclization. Coupling was initiated by a suitable base, such as N,N-diispropylethylamine (DIPEA), sym-collidine or N-methylmorpholine (NMM).Example 4: Biological Data

[0115] The agonist and antagonist activity of B07 at the melanocortin receptors (e.g., MC1 R, MC3R, MC4R, and MC5R) was measured via cAMP accumulation assay, according to the following procedure. Experimental design and execution were conducted by Epics Therapeutics S.A. EuroscreenFast (Bruxelles, Belgium).Compound Handing

[0116] B07 (Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 2) was delivered as powder (1 mg) or 10 mM solutions (100 pl) in 100% DMSO. Powders were solubilized in 100% DMSO at a concentration of 10 mM (master solution) in a defined solvent volume. Serial dilutions were performed from master solution in 100% DMSO to obtain intermediate concentrations 200-, 300- or 400-fold higher than the concentrations to be tested, depending on the assay. The sample was diluted 100-fold in the assay buffer and dispensed in a test plate. Amounts, solvents, and dilutions were estimated based on standard small-molecule drugs. Cell lines used for functional assays are shown in Table 1 .-34-168785213.5Table 1. Cell linesCompound Testing

[0117] B07 was tested for (i) agonist and / or antagonist activity at the human MC3(FAST-0232C) and MC4 (FAST-0233C) receptors, (ii) agonist activity at the human MC1 (FAST-0230C) receptor, and / or (iii) agonist activity at the human MC5 (FAST-0233C) receptor at the following nanomolar concentrations, in duplicate: 0.0001 , 0.001 , 0.01 , 0.03, 0.1 , 0.3, 1 , 10, 100, and 1 ,000.Testing Protocol

[0118] Cyclic AMP (cAMP) Homogenous Time-Resolved Fluorescence (HTRF) assay for Gs coupled receptor:

[0119] CHO-K1 cells expressing recombinant human receptor grown prior to the test in media without antibiotic were detached by gentle flushing with PBS-EDTA (5 mM EDTA), recovered by centrifugation and resuspended in assay buffer (KRH: 5 mM KCI, 1.25 mM MgSO4, 124 mM NaCI, 25 mM HEPES, 13.3 mM Glucose, 1.25 mM KH2PO4, 1.45 mM CaCI2, 0.5 g / l BSA, supplemented with I mM IBMX or 25pM Rolipram).

[0120] Dose response curves were performed in parallel with the reference compounds.

[0121] For agonist test (384well): 5 pl of cells were mixed with 5 pl of the test compound at increasing concentrations and then incubated 30 min at room temperature. After addition of the lysis buffer containing cAMP-d2 and anti-cAMP cryptate detection reagents, plates were incubated 1 -hour at room temperature, and fluorescence ratios were measured according to the manufacturer specification, with the HTRF kit.-35-168785213.5

[0122] For antagonist test (384well): 5 pl of cells were mixed in the wells of an assay plate with 5 pl of a mix of test compound at increasing concentrations and reference agonist for a final concentration corresponding to the historical EC80. The plates were then incubated 30 min at room temperature. After addition of the lysis buffer containing cAMP-d2 and anti-cAMP cryptate detection reagents, plates were incubated 1 -hour at room temperature, and fluorescence ratios were measured according to the manufacturer specification, with the HTRF kit.Quality Control for Compound Testing

[0123] On each day of experimentation and prior to the testing of B07, reference compounds were tested at several concentrations in duplicate (n=2) to obtain a doseresponse curve and an estimated EC50 and / or IC50 values.

[0124] Reference values thus obtained for the test were compared to historical values obtained from the same receptor and used to validate the experimental session.

[0125] A session was considered as valid only if the reference value was found to be within a 0.5 logs interval from the historical value.For replicate determinations, the maximum variability tolerated in the test was of + / -20% around the average of the replicates.Agonist activity of melanocortin analogs on melanocortin 1 receptor and melanocortin 5 receptor

[0126] Administration of B07 activated melanocortin 1 receptor (MC1 R) and melanocortin 5 receptor (MC5R) activity, as measured by cAMP levels (Table 3).Table 2. Dose-response results of B07 and control against the melanocortin 1 receptor (MC1 R) and the melanocortin 5 receptor-36-168785213.5A = > 80% EmaxAgonist activity of melanocortin analogs on melanocortin 3 receptor and melanocortin 4 receptor

[0127] Administration of B07 did not activated melanocortin 3 receptor (MC3R) or melanocortin 4 receptor (MC4R) activity at all, as measured by cAMP levels (Table 3).Table 3. Dose-response results of B07 and control against the melanocortin 3 receptor (MC3R) and melanocortin 4 receptor (MC4R) in agonist mode= > 100 nMC = < 50% EmaxAntagonist activity of melanocortin analogs on melanocortin 3 receptor and melanocortin 4 receptor

[0128] Administration of B07 inhibited melanocortin 3 receptor (MC3R) and melanocortin 4 receptor (MC4R), as measured by cAMP levels (Table 4).Table 4. Dose-response results of B07 and control against the melanocortin 3 receptor (MC3R) and melanocortin 4 receptor (MC4R) in antagonist mode** = 1 -100 nMA = > 80% Emax168785213.5Example 5: Stable Aqueous Compositions of Melanocortin Analog B07

[0129] Three aqueous compositions comprising 50 mg / mL of B07 (Ac-Nle-c[Asp-Pro- dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 2) were tested for stability. The details of each composition are provided in Table 5.Table 5. Aqueous Compositions of 50 mg / mL B07

[0130] The compositions were prepared via the following stepwise compounding process.

[0131] Step 1 : A stock solution comprising the histidine or sodium acetate buffer, the mannitol, and the glacial acetic acid or sodium hydroxide was prepared in excess. The details of the stock solution are provided in Table 6.Table 6. Stock Solution for 50 mg / mL Melanocortin Analog Formulations-38-168785213.5

[0132] The stock solution was prepared by first measuring out 85% of the batch quantity of water to a container. Then, 150 grams of mannitol and 23.28 g of histidine was transferred to the container and the solution was stirred to dissolve the solids. After all solids were dissolved, 7.2 g of sodium hydroxide (equivalent to 7.2 g of sodium hydroxide if a premade solution is being used) was added and stirred to dissolve. Complete dissolution of the added solids in the water was confirmed, then the remaining quantity of water was added while stirring to make up to the final volume. The contents were then mixed to achieve homogenous solution, and the pH of the solution was checked and adjusted to achieve a desired pH based on the desired pH of the final pharmaceutical formulation. For example, for a final pharmaceutical formulation having a pH of about 6.3 (e.g., Formulation 1 ), the pH of the carrier stock solution was adjusted to about 10.5 to about 1 1 .0, whereas for a final formulation having a pH of about 5.3 (e.g., Formulation 2), the pH of the carrier stock solution was adjusted to about 9.3. The stock solution was then kept at room temperature until the next step.

[0133] Step 2: The stock solution and an aqueous solution of the melanocortin analog were combined to provide a stable aqueous composition.

[0134] The aqueous solution of the melanocortin analog was prepared by measuring exactly half the batch quantity of water needed for the composition. With the contents gently mixed, the peptide was added to the water in small quantities and the solution was mixed until all solids dissolved. Foam formation was minimized by maintaining low stirring speeds and avoiding a vortex when possible. After all the melanocortin analog was added, complete dissolution of all solids was confirmed. An exact quantity of the stock solution was then added to the aqueous peptide composition while mixing the contents and the solution was then mixed for another 30 minutes to ensure homogeneity of contents. The pH of the homogenous composition was recorded and adjusted as necessary to achieve the target pH of the composition.

[0135] The appearance, pH, osmolality, and assay / impurity assay of samples at RT (Ambient), 40 °C, and 60 °C conditions at T=0, 3, 7 and 14 days were analyzed to determine the stability of the compositions and are shown in Tables 7-1 1 .Table 7. Formulation 1 Details-39-168785213.5Table 8. Additional Formulation 1 DetailsTable 9. Formulation 2 DetailsTable 9. Additional Formulation 2 DetailsTable 10. Formulation 3 Details-40-168785213.5Table 11. Additional Formulation 3 Details

[0136] Two aqueous compositions comprising 25 mg / mL of B07 (Ac-Nle-c[Asp-Pro- dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 2) were also tested for stability. The compositions were prepared via the following stepwise compounding process.

[0137] Step 1 : A stock solution comprising histidine, mannitol, and glacial acetic acid was prepared in excess. The details of the stock solution are provided in Table 12.Table 12. Stock Solution for 25 mg / mL Melanocortin Analog Formulationsa) Density of 50 mM Histidine with 70.6 mg / mL Mannitol and 0.865 mg / mL Glacial Acetic Acid STOCK solution: 1 .024 g / mL b) Adjust the batch size for 50 mM Histidine with 70.6 mg / mL Mannitol and 0.865 mg / mL Glacial Acetic Acid Stock preparation as required.

[0138] The stock solution was prepared by first measuring out 85% of the batch quantity of water to a container. Then, 21 1.80 grams of mannitol and 23.28 g of histidine was transferred to the container and the solution was stirred to dissolve the solids. After all solids were dissolved, 2.6 g of glacial acetic acid was added and stirred to dissolve. Complete dissolution of the added solids in the water was confirmed, then the remaining -41-168785213.5quantity of water was added while stirring to make up to the final volume. The contents were then mixed for at least 15 minutes to achieve homogenous solution, and the pH of the solution was checked and adjusted to achieve a pH of 6.2 to 6.6. The stock solution was then kept at room temperature until the next step.

[0139] Step 2: The stock solution and an aqueous solution of the melanocortin analog were combined to provide a stable aqueous composition.

[0140] The aqueous solution of the melanocortin analog was prepared by measuring exactly half the batch quantity of water needed for the composition. With the contents gently mixed, the peptide was added to the water in small quantities and the solution was mixed until all solids dissolved. Foam formation was minimized by maintaining low stirring speeds and avoiding a vortex when possible. After all the melanocortin analog was added, complete dissolution of all solids was confirmed. An exact quantity of the stock solution was then added to the aqueous peptide composition while mixing the contents and the solution was then mixed for another 30 minutes to ensure homogeneity of contents. The pH of the homogenous composition was recorded and adjusted as necessary to achieve the target pH of the composition (target pH of 5.3).

[0141] The details of the 25 mg / mL melanocortin analog aqueous formulations are shown in Tables 13 and 14.Table 13. 25 mg / mL Melanocortin Analog Formulation Details - Small Batch-42-168785213.5Table 14. 25 mg / mL Melanocortin Analog Formulation Details - Large Batch

[0142] Additionally, two aqueous compositions comprising 12.5 mg / mL of B07 (Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 2) were tested for stability. The compositions were prepared via the following stepwise compounding process.

[0143] Step 1 : A stock solution comprising histidine, mannitol, and glacial acetic acid was prepared in excess. The details of the stock solution are provided in Table 15.Table 15. Stock Solution for 12.5 mg / mL Melanocortin Analog Formulationsa) Density of 50 mM Histidine with 70.6 mg / mL Mannitol and 0.865 mg / mL Glacial Acetic Acid STOCK solution: 1 .024 g / mL b) Adjust the batch size for 50 mM Histidine with 70.6 mg / mL Mannitol and 0.865 mg / mL Glacial Acetic Acid Stock preparation as required.

[0144] The stock solution was prepared by first measuring out 85% of the batch quantity of water to a container. Then, 225.00 grams of mannitol and 23.28 g of histidine was transferred to the container and the solution was stirred to dissolve the solids. After all solids were dissolved, 6.89 g of glacial acetic acid was added and stirred to dissolve. Complete dissolution of the added solids in the water was confirmed, then the remaining-43-168785213.5quantity of water was added while stirring to make up to the final volume. The contents were then mixed for at least 15 minutes to achieve homogenous solution, and the pH of the solution was checked and adjusted to a pH of 5.5 to 5.9. The stock solution was then kept at room temperature until the next step.

[0145] Step 2: The stock solution and an aqueous solution of the melanocortin analog were combined to provide a stable aqueous composition.

[0146] The aqueous solution of the melanocortin analog was prepared by measuring exactly half the batch quantity of water needed for the composition. With the contents gently mixed, the peptide was added to the water in small quantities and the solution was mixed until all solids dissolved. Foam formation was minimized by maintaining low stirring speeds and avoiding a vortex when possible. After all the melanocortin analog was added, complete dissolution of all solids was confirmed. An exact quantity of the stock solution was then added to the aqueous peptide composition while mixing the contents and the solution was then mixed for another 30 minutes to ensure homogeneity of contents. The pH of the homogenous composition was recorded and adjusted as necessary to achieve the target pH of the composition (target pH of 5.3).

[0147] The details of the 25 mg / mL melanocortin analog aqueous formulations are shown in Tables 16 and 17.Table 16. 12.5 mg / mL Melanocortin Analog Formulation Details - Small Batch-44-168785213.5Table 17. 12.5 mg / mL Melanocortin Analog Formulation Details - Large BatchExample 6: Stability Data for Melanocortin Analog Formulations

[0148] Impurity formation over time in aqueous formulations comprising 50 mg / mL, 25 mg / mL, or 12.5 mg / mL non-naturally occurring melanocortin analog was performed and the corresponding impurity data was assessed. The non-naturally occurring melanocortin receptor analog present in the aqueous compositions had the sequence of: Ac-Nle-c[Asp- Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2; B07). The details of the analysis method and the resulting impurity information are provided below.

[0149] Total impurity regression analysis was used to determine impurity formation in drug products over time. Under this method, impurities are identified in various conditions including (a) real time stability data for drug product maintained at about 2 °C to about 8 °C; (b) accelerated stability data for drug product maintained at 25 °C; and (c) super-accelerated stability data for drug product maintained at 40 °C. The individual impurities identified in drug products subjected to the three stability data conditions for up to nine months are provided in Tables 18-20.Table 18. Individual Impurities in 50 mg B07 Drug Product-45-168785213.5Table 19. Individual Impurities in 25 mg B07 Drug Product-46-168785213.5Table 20. Individual Impurities in 12.5 mg B07 Drug Product-47-168785213.5

[0150] FIGS. 1A-1 F show linear regression models of the total impurity data for samples maintained at 5 °C (FIGS. 1 A-1 C) and 25 °C (FIGS. 1 D-1 F) over the course of nine months.

[0151] Additionally, particulate, osmolality, and pH were tested at the same time points that the total impurity analysis was performed for each sample. The data are shown in Tables 21 -23.Table 21. Particulate, Osmolality, and pH of 50 mg B07 Drug Product-48-168785213.5Table 22. Particulate, Osmolality, and pH of 25 mg B07 Drug ProductTable 23. Particulate, Osmolality, and pH of 12.5 mg B07 Drug ProductExample 7: Additional Stable Aqueous Compositions of Melanocortin Analog B07

[0152] Eight aqueous formulations of B07 including sodium acetate as a buffer, as opposed to histidine, were prepared and tested for stability. Table 24 provides details of the formulations.Table 24. Second Generation Stable Aqueous Injectable Formulations of B07-49-168785213.5

[0153] The formulations were maintained at various different conditions and tested for osmolality, pH, purity, and particulate matter (via dynamic light scattering (DLS) radius) at various time points for up to four months. The results are shown in Tables 25-30.Table 25. Osmolality (mOSm / kg) of Formulations 1 -8Table 26. pH of Formulations 1 -8-50-168785213.5Table 27. HPLC Purity (%) of Formulations 1-8Table 28. HPLC Assay Concentration of B07 (mg / mL) of Formulations 1-8Table 29. DLS Radius (nm) of Formulations 1-8-51-168785213.5*Microscopic aggregation was observed at 1 and 2 months for F1 stored at 5 °C and at 1 month for F1 stored at 25 °C and 40 °C**Microscopic aggregation was observed at 2 months for F2 stored at all three temperatures.

[0154] An impurity peak at around 12.2 minutes of the HPLC assay was observed across all samples including baseline. The area of the peak for each formulation at all conditions is provided below in Table 30.Table 30. Area of impurity (area %) at around 12.2 min of HPLC Assay of Formulations 1 -8

[0155] Based on the results of the stability analysis for Formulations 1 -8, Formulations 9-20D were prepared to achieve even greater stability of the injectable formulation comprising melanocortin analogs (Table 31 ).Table 31. Third Generation Stable Aqueous Injectable Formulations of 50 mg / mL B07-52-168785213.5

[0156] Formulations 9-19 will be maintained at 5 °C, 25 °C, and 40 °C and tested for pH, osmolality, purity, and particulate matter at various time points for up to six months to determine the stability of each formulation.Additional Embodiments

[0157] Various embodiments of the present technology are set forth below in paragraphs

[0158] to

[0267] :

[0158] 1. A pharmaceutical composition having a pH of about 4.0 to about 6.5, comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), an osmolyte, a pH adjusting agent, and a buffer, wherein c in the sequence represents cyclization via a lactam bond.

[0159] 2. A pharmaceutical composition having a pH of about 4.0 to about 6.5, comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), an osmolyte, a pH adjusting agent, and a buffer, wherein a cationic residue of the non-naturally occurring melanocortin analog is protonated and the c in the sequence represents cyclization via a lactam bond.-53-168785213.5

[0160] 3. A pharmaceutical formulation comprising an aqueous peptide composition and a pharmaceutically acceptable carrier composition, the aqueous peptide composition comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond and the pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer.

[0161] 4. A pharmaceutical formulation having a pH of about 4.0 to about 6.5 and comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, the formulation produced by: combining an aqueous peptide composition comprising the melanocortin analog, and a pharmaceutically acceptable carrier composition having a pH of about 5.5 to about 11 .0 comprising an osmolyte, a pH adjusting agent, and a buffer; and preventing aggregation of the analog by maintaining a local pH of the pharmaceutical formulation such that no local region of the analog has a pH equal to the isoelectric point of the analog while the aqueous peptide composition and the pharmaceutically acceptable carrier composition are being combined.

[0162] 5. A pharmaceutical formulation having a pH of about 4.0 to about 6.5 and comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, the formulation produced by combining an aqueous peptide composition comprising the non-naturally occurring melanocortin analog, and a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer and having a pH of about 5.5 to about 1 1 .0.

[0163] 6. The pharmaceutical composition or pharmaceutical formulation of embodiment any one of embodiments 1 -4, wherein the pharmaceutical composition or the pharmaceutical formulation has a pH of about 4.0 to about 6.5.-54-168785213.5

[0164] 7. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -6, wherein the buffer is present at a concentration of about 10 m / l to about 200 mM.

[0165] 8. The pharmaceutical composition or the pharmaceutical formulation of embodiment 7, wherein the buffer is present at a concentration of about 5 mM to about 100 mM.

[0166] 9. The pharmaceutical composition or the pharmaceutical formulation of embodiment 7 or 8, wherein the buffer is present at a concentration of about 10 mM to about 25 mM.

[0167] 10. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -9, wherein the buffer comprises one or more of histidine, Tris, arginine, MES, ADA, PIPES, ACES, MOPSO, MOPS, Bes, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, HEPPS, TPAS, acetoamidoglycine, glycinamide, glycylglycine, tricine, and bicine.

[0168] 11. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -10, wherein the buffer comprises histidine.

[0169] 12. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -9, wherein the buffer comprises sodium acetate

[0170] 13. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -12, wherein the non-naturally occurring melanocortin analog is present in an amount of about 10 mg / mL to about 130 mg / mL.

[0171] 14. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -13, wherein the non-naturally occurring melanocortin analog is present in an amount of about 10 mg / mL to about 50 mg / mL.

[0172] 15. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -14, wherein the non-naturally occurring melanocortin analog is present in an amount of about 12.5 mg / mL.-55-168785213.5

[0173] 16. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -14, wherein the non-naturally occurring melanocortin analog is present in an amount of about 25 mg / mL.

[0174] 17. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -14, wherein the non-naturally occurring melanocortin analog is present in an amount of about 50 mg / mL.

[0175] 18. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -17, wherein the osmolyte is selected from the group consisting of a sugar or derivative thereof, a glycerol, a glycerin, a gelatin, a propylene glycol, a propylparaben, PVP, PLGA, and PEG.

[0176] 19. The pharmaceutical composition or pharmaceutical formulation of embodiment 18, wherein the sugar or the derivative thereof is a sugar alcohol.

[0177] 20. The pharmaceutical composition or pharmaceutical formulation of embodiment 19, where the sugar alcohol is mannitol.

[0178] 21. The pharmaceutical composition or pharmaceutical formulation of embodiment 20, wherein the mannitol is present in an amount of about 10 mg / mL to about 50 mg / mL.

[0179] 22. The pharmaceutical composition or pharmaceutical formulation of embodiment 20 or 21 , wherein the mannitol is present in an amount of about 25 mg / mL.

[0180] 23. The pharmaceutical composition or pharmaceutical formulation of embodiment 20 or 21 , wherein the mannitol is present in an amount of about 27.5 mg / mL.

[0181] 24. The pharmaceutical composition or pharmaceutical formulation of embodiment 20 or 21 , wherein the mannitol is present in an amount of about 35 mg / mL.

[0182] 25. The pharmaceutical composition or pharmaceutical formulation of embodiment 20 or 21 , wherein the mannitol is present in an amount of about 37.5 mg / mL.

[0183] 26. The pharmaceutical composition of formulation of any one of embodiments 1 -25, wherein the pH adjusting agent is present in an amount of about 0.01 mg / mL to about 10 mg / mL.-56-168785213.5

[0184] 27. The pharmaceutical composition or formulation of any one of embodiments 1 -26, wherein the pH adjusting agent is a base.

[0185] 28. The pharmaceutical composition or formulation of embodiment 27, wherein the base is sodium hydroxide.

[0186] 29. The pharmaceutical composition or formulation of embodiment 28, wherein the sodium hydroxide is present in an amount of about 0.01 mg / mL to about 5.0 mg / L.

[0187] 30. The pharmaceutical composition or formulation of embodiment 28 or29, wherein the sodium hydroxide is present in an amount of about 0.4 mg / mL.

[0188] 31. The pharmaceutical composition or formulation of embodiment 28 or29, wherein the sodium hydroxide is present in an amount of about 1 .2 mg / mL.

[0189] 32. The pharmaceutical composition or formulation of embodiment 28 or29, wherein the sodium hydroxide is present in an amount of about 1 .6 mg / mL.

[0190] 33. The pharmaceutical composition or formulation of embodiment 27, wherein the base is histidine.

[0191] 34. The pharmaceutical composition or formulation of embodiment 33, wherein the histidine is present in an amount of about 0.01 mg / mL to about 5.0 mg / mL.

[0192] 35. The pharmaceutical composition or formulation of any one of embodiments 1 -25, wherein the pH adjusting agent is an acid.

[0193] 36. The pharmaceutical composition or formulation of embodiment 35, wherein the acid is glacial acetic acid.

[0194] 37. The pharmaceutical composition or formulation of embodiment 36, wherein the glacial acetic acid is present in an amount of about 0.01 mg / mL to about 5.0 mg / mL.

[0195] 38. The pharmaceutical composition or formulation of any one of embodiments 1 -37, further comprising a surfactant.-57-168785213.5

[0196] 39. The pharmaceutical composition or formulation of embodiment 38, wherein the surfactant is selected from an alkyl-carboxylate-fatty acid salt, a sulfate, a sulfonate, a phosphate ester, a quaternary ammonium surfactant, a pyridinium surfactant, a polyol ester, a polyoxyethylene ester, a poloxamer, a phospholipid, and a betaine.

[0197] 40. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -39, further comprising substantially no calcium ions.

[0198] 41. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -40, further comprising substantially no metal ions.

[0199] 42. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -41 , having an osmolality of about 250 mOsm / kg to about 600 mOsm / kg.

[0200] 43. The pharmaceutical composition or the pharmaceutical formulation of any one of embodiments 1 -42, having an osmolality of about 270 mOsm / kg to about 330 mOsm / kg.

[0201] 44. The pharmaceutical composition or formulation of any one of embodiments 1 -43, further comprising a preservative.

[0202] 45. The pharmaceutical composition or formulation of embodiment 44, wherein the preservative is selected from the group consisting of m-cresol, phenol, and benzyl alcohol.

[0203] 46. The pharmaceutical composition or formulation of embodiment 44 or45, wherein the preservative is present in an amount of about 0.1 wt% to about 1.0 wt%, relative to a total weight of the composition or formulation.

[0204] 47. A pharmaceutical composition having a pH of about 5.3, the composition comprising: a non-naturally occurring melanocortin analog in an amount of 12.5 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal- dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond; mannitol in an amount of about 30 mg / mL to about 40 mg / mL;-58-168785213.5an acid in an amount of about 0.2 mg / mL to about 1 .6 mg / mL; and histidine at a concentration of about 25 mM.

[0205] 48. The pharmaceutical composition of embodiment 47, wherein the mannitol is present in an amount of about 37.5 mg / mL.

[0206] 49. The pharmaceutical composition of embodiment 47 or 48, wherein the acid is glacial acetic acid.

[0207] 50. A pharmaceutical composition having a pH of about 5.3, the composition comprising: a non-naturally occurring melanocortin analog in an amount of 25 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal- dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond; mannitol in an amount of about 30 mg / mL to about 40 mg / mL; an acid in an amount of about 0.2 mg / mL to about 1 .6 mg / mL; and histidine at a concentration of about 25 mM.

[0208] 51 . The pharmaceutical composition of embodiment 50, wherein the mannitol is present in an amount of about 35 mg / mL.

[0209] 52. The pharmaceutical composition of embodiment 50 or 51 , wherein the acid is glacial acetic acid.

[0210] 53. A pharmaceutical composition having a pH of about 4.0 to about 6.5, the composition comprising: a non-naturally occurring melanocortin analog in an amount of 50 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal- dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond; mannitol in an amount of about 25 mg / mL to about 35 mg / mL; a base in an amount of about 0.1 mg / mL to about 2.0 mg / mL; and a buffer at a concentration of about 25 mM.-59-168785213.5

[0211] 54. The pharmaceutical composition of embodiment 53, wherein the mannitol is present in an amount of about 25 mg / mL.

[0212] 55. The pharmaceutical composition of embodiment 53, wherein the mannitol is present in an amount of about 27.5 mg / mL.

[0213] 56. The pharmaceutical composition of any one of embodiments 53-55, wherein the base is sodium hydroxide.

[0214] 57. The pharmaceutical composition of embodiment 56, wherein the sodium hydroxide is present in an amount of 0.4 mg / mL, the buffer is histidine, and the pH of the pharmaceutical composition is about 5.3.

[0215] 58. The pharmaceutical composition of embodiment 56, wherein the sodium hydroxide is present in an amount of 1 .2 mg / mL, the buffer is histidine, and the pH of the pharmaceutical composition is about 6.3.

[0216] 59. The pharmaceutical composition of embodiment 56, wherein the sodium hydroxide is present in an amount of 0.5 mg / mL, the buffer is sodium acetate, and the pH of the pharmaceutical composition is about 5.0.

[0217] 60. A method of producing a pharmaceutical composition of a non-naturally occurring melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp- Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, the method comprising combining an aqueous peptide composition of the non-naturally occurring melanocortin analog with a pharmaceutically acceptable carrier composition having a pH of about 5.5 to about 11 .0 and comprising an osmolyte, a pH adjusting agent, and a buffer, wherein the pharmaceutical composition has a pH of about 4.0 to about 6.5.

[0218] 61. A method of producing a pharmaceutical formulation having a pH of about 4.0 to about 6.5, the method comprising: combining an aqueous peptide composition of a non-naturally occurring melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, with a pharmaceutically-60-168785213.5acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer; and preventing aggregation of the analog by maintaining a local pH of the pharmaceutical composition such that no local region has a pH that is equal to the isoelectric point of the analog while the aqueous peptide composition and the pharmaceutically acceptable carrier composition are being combined.

[0219] 62. A method of producing a pharmaceutical formulation of a non-naturally occurring melanocortin analog having a pH of about 4.0 to about 6.5, the method comprising combining an aqueous peptide composition of the non-naturally occurring melanocortin analog with a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer, wherein the non-naturally occurring melanocortin analog has the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond and the pharmaceutically acceptable carrier composition has a pH of about 5.5 to about 1 1 .0.

[0220] 63. A method of producing a pharmaceutical formulation of a non-naturally occurring melanocortin analog, the method comprising: providing a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a histidine buffer; dissolving a non-naturally melanocortin having the sequence: Ac-Nle-c[Asp-Pro- dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond in water to provide an aqueous peptide composition; and adding the pharmaceutically acceptable carrier composition to the aqueous peptide composition while mixing to maintain dissolution of the non-naturally occurring melanocortin analog, wherein the pharmaceutical formulation has a pH of about 4.0 to about 6.5.

[0221] 64. The method of any one of embodiment 60-62, wherein combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition comprises adding the pharmaceutically acceptable carrier composition to the aqueous peptide composition.-61 -168785213.5

[0222] 65. The method of any one of embodiment 60-62, wherein combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition comprises adding the aqueous peptide composition to the pharmaceutically acceptable carrier composition.

[0223] 66. The method of any one of embodiments 60-65, wherein the pharmaceutical composition contains substantially no precipitated non-naturally occurring melanocortin analog.

[0224] 67. The method of any one of embodiments 60-66, further comprising, after combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition, stirring the combination for a period of time sufficient to achieve a stable homogenous composition.

[0225] 68. The method of embodiment 67, wherein the period of time sufficient to achieve a stable homogenous composition is about 10 minutes to about 24 hours.

[0226] 69. The method of any one of embodiments 60-68, further comprising, after combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition, adjusting a pH of the combination to achieve a pH of about 5.0 to about 6.5.

[0227] 70. The method of embodiment 69, wherein adjusting the pH of the combination comprises adding sodium hydroxide, histidine, or glacial acetic acid to the combination.

[0228] 71. The method of any one of embodiments 60-62, wherein the buffer comprises one or more of histidine, Tris, arginine, MES, ADA, PIPES, ACES, MOPSO, MOPS, Bes, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, HEPPS, TPAS, acetoamidoglycine, glycinamide, glycylglycine, tricine, and bicine.

[0229] 72. The method of any one of embodiments 60-71 , wherein the buffer comprises histidine.

[0230] 73. The method of any one of embodiments 60-70, wherein the buffer comprises sodium acetate.-62-168785213.5

[0231] 74. The method of any one of embodiments 60-73, wherein the buffer is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 200 mM.

[0232] 75. The method of embodiment 74, wherein the buffer is present in the pharmaceutical composition or formulation at a concentration of about 5 mM to about 100 mM.

[0233] 76. The method of embodiment 74 or 72, wherein the buffer is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 25 mM.

[0234] 77. The method of any one of embodiments 60-76, wherein the non- naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 10 mg / mL to about 130 mg / mL.

[0235] 78. The method of any one of embodiments 60-77, wherein the non- naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation at a concentration of about in an amount of about 10 mg / mL to about 50 mg / mL.

[0236] 79. The method of any one of embodiments 60-78, wherein the non- naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 12.5 mg / mL.

[0237] 80. The method of any one of embodiments 60-78, wherein the non- naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 25 mg / mL.

[0238] 81 . The method of any one of embodiments 60-78, wherein the non- naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 50 mg / mL.

[0239] 82. The method of any one of embodiments 60-81 , wherein the osmolyte is selected from the group consisting of a sugar or derivative thereof, a glycerol, a glycerin, a gelatin, a propylene glycol, a propylparaben, PVP, PLGA, and PEG.-63-168785213.5

[0240] 83. The method of embodiment 82, wherein the sugar or the derivative thereof is a sugar alcohol.

[0241] 84. The method of embodiment 83, wherein the sugar alcohol is mannitol.

[0242] 85. The method of embodiment 84, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 20 mg / mL to about 40 mg / mL.

[0243] 86. The method of embodiment 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 25 mg / mL.

[0244] 87. The method of embodiment 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 27.5 mg / mL.

[0245] 88. The method of embodiment 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 35 mg / mL.

[0246] 89. The method of embodiment 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 37.5 mg / mL.

[0247] 90. The method of any one of embodiments 60-89, wherein the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 10 mg / mL.

[0248] 91. The method of any one of embodiments 60-90, wherein the pH adjusting agent is a base.

[0249] 92. The method of embodiment 91 , wherein the base is sodium hydroxide.

[0250] 93. The method of embodiment 92, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / L.

[0251] 94. The method of embodiment 92 or 93, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.4 mg / mL.

[0252] 95. The method of embodiment 92 or 93, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .2 mg / mL.-64-168785213.5

[0253] 96. The method of embodiment 92 or 93, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .6 mg / mL.

[0254] 97. The method of embodiment 91 , wherein the base is histidine.

[0255] 98. The method of embodiment 97, wherein the histidine is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / L.

[0256] 99. The method of any one of embodiments 1 -89, wherein the pH adjusting agent is an acid.

[0257] 100. The method of embodiment 99, wherein the acid is glacial acetic acid.

[0258] 101. The method of embodiment 100, wherein the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / mL.

[0259] 102. The method of any one of embodiments 60-101 , wherein the pharmaceutical composition or formulation further comprises a surfactant.

[0260] 103. The method of embodiment 102, wherein the surfactant is selected from an alkyl-carboxylate-fatty acid salt, a sulfate, a sulfonate, a phosphate ester, a quaternary ammonium surfactant, a pyridinium surfactant, a polyol ester, a polyoxyethylene ester, a poloxamer, a phospholipid, and a betaine.

[0261] 104. The method of any one of embodiments 60-103, wherein the pharmaceutically acceptable carrier composition comprises substantially no calcium ions.

[0262] 105. The method any one of embodiments 60-104, wherein the pharmaceutically acceptable carrier composition comprises substantially no metal ions.

[0263] 106. The method of any one of embodiments 60-105, wherein the pharmaceutical composition or formulation has an osmolality of about 250 mOsm / kg to about 600 mOsm / kg.

[0264] 107. The method of any one of embodiments 60-106, wherein the pharmaceutical composition or formulation has an osmolality of about 270 mOsm / kg to about 330 mOsm / kg.-65-168785213.5

[0265] 108. The method of any one of embodiments 60-107, wherein the pharmaceutical composition or formulation further comprises a preservative.

[0266] 109. The method of embodiment 108, wherein the preservative is selected from the group consisting of m-cresol, phenol, and benzyl alcohol.

[0267] 110. The method of embodiment 108 or 109, wherein the preservative is present in the pharmaceutical composition or formulation in an amount of about 0.1 wt% to about 1 .0 wt%, relative to a total weight of the composition or formulation.

[0268] From the foregoing, it will be appreciated that specific embodiments of the invention have been described herein for purposes of illustration, but that various modifications may be made without deviating from the scope of the invention. Accordingly, the invention is not limited except as by the appended claims.-66-168785213.5

Claims

CLAIMS l / We claim:

1. A pharmaceutical composition having a pH of about 4.0 to about 6.5, comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), an osmolyte, a pH adjusting agent, and a buffer, wherein c in the sequence represents cyclization via a lactam bond.

2. A pharmaceutical composition having a pH of about 4.0 to about 6.5, comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), an osmolyte, a pH adjusting agent, and a buffer, wherein a cationic residue of the non-naturally occurring melanocortin analog is protonated and the c in the sequence represents cyclization via a lactam bond.

3. A pharmaceutical formulation comprising an aqueous peptide composition and a pharmaceutically acceptable carrier composition, the aqueous peptide composition comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond and the pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer.

4. A pharmaceutical formulation having a pH of about 4.0 to about 6.5 and comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, the formulation produced by:-67-168785213.5combining an aqueous peptide composition comprising the melanocortin analog, and a pharmaceutically acceptable carrier composition having a pH of about 5.5 to about 11 .0 comprising an osmolyte, a pH adjusting agent, and a buffer; and preventing aggregation of the analog by maintaining a local pH of the pharmaceutical formulation such that no local region of the analog has a pH equal to the isoelectric point of the analog while the aqueous peptide composition and the pharmaceutically acceptable carrier composition are being combined.

5. A pharmaceutical formulation having a pH of about 4.0 to about 6.5 and comprising a non-naturally occurring melanocortin analog having the sequence: Ac-Nle- c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, the formulation produced by combining an aqueous peptide composition comprising the non-naturally occurring melanocortin analog, and a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer and having a pH of about 5.5 to about 1 1 .0.

6. The pharmaceutical composition or pharmaceutical formulation of claim any one of claims 1 -4, wherein the pharmaceutical composition or the pharmaceutical formulation has a pH of about 4.0 to about 6.5.

7. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -6, wherein the buffer is present at a concentration of about 10 mM to about 200 mM.

8. The pharmaceutical composition or the pharmaceutical formulation of claim 7, wherein the buffer is present at a concentration of about 5 mM to about 100 mM.

9. The pharmaceutical composition or the pharmaceutical formulation of claim 7 or 8, wherein the buffer is present at a concentration of about 10 mM to about 25 mM.-68-168785213.

510. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -9, wherein the buffer comprises one or more of histidine, Tris, arginine, MES, ADA, PIPES, ACES, MOPSO, MOPS, Bes, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, HEPPS, TPAS, acetoamidoglycine, glycinamide, glycylglycine, tricine, and bicine.1 1 . The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -10, wherein the buffer comprises histidine.

12. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -9, wherein the buffer comprises sodium acetate13. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -12, wherein the non-naturally occurring melanocortin analog is present in an amount of about 10 mg / mL to about 130 mg / mL.

14. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -13, wherein the non-naturally occurring melanocortin analog is present in an amount of about 10 mg / mL to about 50 mg / mL.

15. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -14, wherein the non-naturally occurring melanocortin analog is present in an amount of about 12.5 mg / mL.

16. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -14, wherein the non-naturally occurring melanocortin analog is present in an amount of about 25 mg / mL.

17. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -14, wherein the non-naturally occurring melanocortin analog is present in an amount of about 50 mg / mL.-69-168785213.

518. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -17, wherein the osmolyte is selected from the group consisting of a sugar or derivative thereof, a glycerol, a glycerin, a gelatin, a propylene glycol, a propylparaben, PVP, PLGA, and PEG.

19. The pharmaceutical composition or pharmaceutical formulation of claim 18, wherein the sugar or the derivative thereof is a sugar alcohol.

20. The pharmaceutical composition or pharmaceutical formulation of claim 19, where the sugar alcohol is mannitol.21 . The pharmaceutical composition or pharmaceutical formulation of claim 20, wherein the mannitol is present in an amount of about 10 mg / mL to about 50 mg / mL.

22. The pharmaceutical composition or pharmaceutical formulation of claim 20 or 21 , wherein the mannitol is present in an amount of about 25 mg / mL.

23. The pharmaceutical composition or pharmaceutical formulation of claim 20 or 21 , wherein the mannitol is present in an amount of about 27.5 mg / mL.

24. The pharmaceutical composition or pharmaceutical formulation of claim 20 or 21 , wherein the mannitol is present in an amount of about 35 mg / mL.

25. The pharmaceutical composition or pharmaceutical formulation of claim 20 or 21 , wherein the mannitol is present in an amount of about 37.5 mg / mL.

26. The pharmaceutical composition of formulation of any one of claims 1 -25, wherein the pH adjusting agent is present in an amount of about 0.01 mg / mL to about 10 mg / mL.-70-168785213.

527. The pharmaceutical composition or formulation of any one of claims 1 -26, wherein the pH adjusting agent is a base.

28. The pharmaceutical composition or formulation of claim 27, wherein the base is sodium hydroxide.

29. The pharmaceutical composition or formulation of claim 28, wherein the sodium hydroxide is present in an amount of about 0.01 mg / mL to about 5.0 mg / L.

30. The pharmaceutical composition or formulation of claim 28 or 29, wherein the sodium hydroxide is present in an amount of about 0.4 mg / mL.31 . The pharmaceutical composition or formulation of claim 28 or 29, wherein the sodium hydroxide is present in an amount of about 1 .2 mg / mL.

32. The pharmaceutical composition or formulation of claim 28 or 29, wherein the sodium hydroxide is present in an amount of about 1 .6 mg / mL.

33. The pharmaceutical composition or formulation of claim 27, wherein the base is histidine.

34. The pharmaceutical composition or formulation of claim 33, wherein the histidine is present in an amount of about 0.01 mg / mL to about 5.0 mg / mL.

35. The pharmaceutical composition or formulation of any one of claims 1 -25, wherein the pH adjusting agent is an acid.

36. The pharmaceutical composition or formulation of claim 35, wherein the acid is glacial acetic acid.-71 -168785213.

537. The pharmaceutical composition or formulation of claim 36, wherein the glacial acetic acid is present in an amount of about 0.01 mg / mL to about 5.0 mg / mL.

38. The pharmaceutical composition or formulation of any one of claims 1 -37, further comprising a surfactant.

39. The pharmaceutical composition or formulation of claim 38, wherein the surfactant is selected from an alkyl-carboxylate-fatty acid salt, a sulfate, a sulfonate, a phosphate ester, a quaternary ammonium surfactant, a pyridinium surfactant, a polyol ester, a polyoxyethylene ester, a poloxamer, a phospholipid, and a betaine.

40. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -39, further comprising substantially no calcium ions.41 . The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -40, further comprising substantially no metal ions.

42. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -41 , having an osmolality of about 250 mOsm / kg to about 600 mOsm / kg.

43. The pharmaceutical composition or the pharmaceutical formulation of any one of claims 1 -42, having an osmolality of about 270 mOsm / kg to about 330 mOsm / kg.

44. The pharmaceutical composition or formulation of any one of claims 1 -43, further comprising a preservative.

45. The pharmaceutical composition or formulation of claim 44, wherein the preservative is selected from the group consisting of m-cresol, phenol, and benzyl alcohol.-72-168785213.

546. The pharmaceutical composition or formulation of claim 44 or 45, wherein the preservative is present in an amount of about 0.1 wt% to about 1 .0 wt%, relative to a total weight of the composition or formulation.

47. A pharmaceutical composition having a pH of about 5.3, the composition comprising: a non-naturally occurring melanocortin analog in an amount of 12.5 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal- dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond; mannitol in an amount of about 30 mg / mL to about 40 mg / mL; an acid in an amount of about 0.2 mg / mL to about 1 .6 mg / mL; and histidine at a concentration of about 25 mM.

48. The pharmaceutical composition of claim 47, wherein the mannitol is present in an amount of about 37.5 mg / mL.

49. The pharmaceutical composition of claim 47 or 48, wherein the acid is glacial acetic acid.

50. A pharmaceutical composition having a pH of about 5.3, the composition comprising: a non-naturally occurring melanocortin analog in an amount of 25 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal- dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond; mannitol in an amount of about 30 mg / mL to about 40 mg / mL; an acid in an amount of about 0.2 mg / mL to about 1 .6 mg / mL; and histidine at a concentration of about 25 mM.51 . The pharmaceutical composition of claim 50, wherein the mannitol is present in an amount of about 35 mg / mL.-73-168785213.

552. The pharmaceutical composition of claim 50 or 51 , wherein the acid is glacial acetic acid.

53. A pharmaceutical composition having a pH of about 4.0 to about 6.5, the composition comprising: a non-naturally occurring melanocortin analog in an amount of 50 mg / mL, the melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal- dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond; mannitol in an amount of about 25 mg / mL to about 35 mg / mL; a base in an amount of about 0.1 mg / mL to about 2.0 mg / mL; and a buffer at a concentration of about 25 mM.

54. The pharmaceutical composition of claim 53, wherein the mannitol is present in an amount of about 25 mg / mL.

55. The pharmaceutical composition of claim 53, wherein the mannitol is present in an amount of about 27.5 mg / mL.

56. The pharmaceutical composition of any one of claims 53-55, wherein the base is sodium hydroxide.

57. The pharmaceutical composition of claim 56, wherein the sodium hydroxide is present in an amount of 0.4 mg / mL, the buffer is histidine, and the pH of the pharmaceutical composition is about 5.3.

58. The pharmaceutical composition of claim 56, wherein the sodium hydroxide is present in an amount of 1 .2 mg / mL, the buffer is histidine, and the pH of the pharmaceutical composition is about 6.3.-74-168785213.

559. The pharmaceutical composition of claim 56, wherein the sodium hydroxide is present in an amount of 0.5 mg / mL, the buffer is sodium acetate, and the pH of the pharmaceutical composition is about 5.0.

60. A method of producing a pharmaceutical composition of a non-naturally occurring melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp- Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, the method comprising combining an aqueous peptide composition of the non-naturally occurring melanocortin analog with a pharmaceutically acceptable carrier composition having a pH of about 5.5 to about 11 .0 and comprising an osmolyte, a pH adjusting agent, and a buffer, wherein the pharmaceutical composition has a pH of about 4.0 to about 6.5.61 . A method of producing a pharmaceutical formulation having a pH of about 4.0 to about 6.5, the method comprising: combining an aqueous peptide composition of a non-naturally occurring melanocortin analog having the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond, with a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer; and preventing aggregation of the analog by maintaining a local pH of the pharmaceutical composition such that no local region has a pH that is equal to the isoelectric point of the analog while the aqueous peptide composition and the pharmaceutically acceptable carrier composition are being combined.

62. A method of producing a pharmaceutical formulation of a non-naturally occurring melanocortin analog having a pH of about 4.0 to about 6.5, the method comprising combining an aqueous peptide composition of the non-naturally occurring melanocortin analog with a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a buffer, wherein the non-naturally occurring melanocortin analog has the sequence: Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2),-75-168785213.5wherein c represents cyclization via a lactam bond and the pharmaceutically acceptable carrier composition has a pH of about 5.5 to about 1 1 .0.

63. A method of producing a pharmaceutical formulation of a non-naturally occurring melanocortin analog, the method comprising: providing a pharmaceutically acceptable carrier composition comprising an osmolyte, a pH adjusting agent, and a histidine buffer; dissolving a non-naturally melanocortin having the sequence: Ac-Nle-c[Asp-Pro- dNal(2’)-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 2), wherein c represents cyclization via a lactam bond in water to provide an aqueous peptide composition; and adding the pharmaceutically acceptable carrier composition to the aqueous peptide composition while mixing to maintain dissolution of the non-naturally occurring melanocortin analog, wherein the pharmaceutical formulation has a pH of about 4.0 to about 6.5.

64. The method of any one of claim 60-62, wherein combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition comprises adding the pharmaceutically acceptable carrier composition to the aqueous peptide composition.

65. The method of any one of claim 60-62, wherein combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition comprises adding the aqueous peptide composition to the pharmaceutically acceptable carrier composition.

66. The method of any one of claims 60-65, wherein the pharmaceutical composition contains substantially no precipitated non-naturally occurring melanocortin analog.

67. The method of any one of claims 60-66, further comprising, after combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition,-76-168785213.5stirring the combination for a period of time sufficient to achieve a stable homogenous composition.

68. The method of claim 67, wherein the period of time sufficient to achieve a stable homogenous composition is about 10 minutes to about 24 hours.

69. The method of any one of claims 60-68, further comprising, after combining the aqueous peptide composition and the pharmaceutically acceptable carrier composition, adjusting a pH of the combination to achieve a pH of about 5.0 to about 6.5.

70. The method of claim 69, wherein adjusting the pH of the combination comprises adding sodium hydroxide, histidine, or glacial acetic acid to the combination.71 . The method of any one of claims 60-62, wherein the buffer comprises one or more of histidine, Tris, arginine, MES, ADA, PIPES, ACES, MOPSO, MOPS, Bes, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, HEPPS, TPAS, acetoamidoglycine, glycinamide, glycylglycine, tricine, and bicine.

72. The method of any one of claims 60-71 , wherein the buffer comprises histidine.

73. The method of any one of claims 60-70, wherein the buffer comprises sodium acetate.

74. The method of any one of claims 60-73, wherein the buffer is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 200 mM.

75. The method of claim 74, wherein the buffer is present in the pharmaceutical composition or formulation at a concentration of about 5 mM to about 100 mM.-77-168785213.

576. The method of claim 74 or 72, wherein the buffer is present in the pharmaceutical composition or formulation at a concentration of about 10 mM to about 25 mM.

77. The method of any one of claims 60-76, wherein the non-naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 10 mg / mL to about 130 mg / mL.

78. The method of any one of claims 60-77, wherein the non-naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation at a concentration of about in an amount of about 10 mg / mL to about 50 mg / mL.

79. The method of any one of claims 60-78, wherein the non-naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 12.5 mg / mL.

80. The method of any one of claims 60-78, wherein the non-naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 25 mg / mL.81 . The method of any one of claims 60-78, wherein the non-naturally occurring melanocortin analog is present in the pharmaceutical composition or formulation in an amount of about 50 mg / mL.

82. The method of any one of claims 60-81 , wherein the osmolyte is selected from the group consisting of a sugar or derivative thereof, a glycerol, a glycerin, a gelatin, a propylene glycol, a propylparaben, PVP, PLGA, and PEG.

83. The method of claim 82, wherein the sugar or the derivative thereof is a sugar alcohol.-78-168785213.

584. The method of claim 83, wherein the sugar alcohol is mannitol.

85. The method of claim 84, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 20 mg / mL to about 40 mg / mL.

86. The method of claim 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 25 mg / mL.

87. The method of claim 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 27.5 mg / mL.

88. The method of claim 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 35 mg / mL.

89. The method of claim 85, wherein the mannitol is present in the pharmaceutical composition or formulation in an amount of about 37.5 mg / mL.

90. The method of any one of claims 60-89, wherein the pH adjusting agent is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 10 mg / mL.91 . The method of any one of claims 60-90, wherein the pH adjusting agent is a base.

92. The method of claim 91 , wherein the base is sodium hydroxide.

93. The method of claim 92, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / L.-79-168785213.

594. The method of claim 92 or 93, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 0.4 mg / mL.

95. The method of claim 92 or 93, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .2 mg / mL.

96. The method of claim 92 or 93, wherein the sodium hydroxide is present in the pharmaceutical composition or formulation in an amount of about 1 .6 mg / mL.

97. The method of claim 91 , wherein the base is histidine.

98. The method of claim 97, wherein the histidine is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / L.

99. The method of any one of claims 1 -89, wherein the pH adjusting agent is an acid.

100. The method of claim 99, wherein the acid is glacial acetic acid.

101. The method of claim 100, wherein the glacial acetic acid is present in the pharmaceutical composition or formulation in an amount of about 0.01 mg / mL to about 5.0 mg / mL.

102. The method of any one of claims 60-101 , wherein the pharmaceutical composition or formulation further comprises a surfactant.

103. The method of claim 102, wherein the surfactant is selected from an alkylcarboxylate-fatty acid salt, a sulfate, a sulfonate, a phosphate ester, aquaternary ammonium surfactant, a pyridinium surfactant, a polyol ester, a polyoxyethylene ester, a poloxamer, a phospholipid, and a betaine.-80-168785213.5104. The method of any one of claims 60-103, wherein the pharmaceutically acceptable carrier composition comprises substantially no calcium ions.

105. The method any one of claims 60-104, wherein the pharmaceutically acceptable carrier composition comprises substantially no metal ions.

106. The method of any one of claims 60-105, wherein the pharmaceutical composition or formulation has an osmolality of about 250 mOsm / kg to about 600 mOsm / kg.

107. The method of any one of claims 60-106, wherein the pharmaceutical composition or formulation has an osmolality of about 270 mOsm / kg to about 330 mOsm / kg.

108. The method of any one of claims 60-107, wherein the pharmaceutical composition or formulation further comprises a preservative.

109. The method of claim 108, wherein the preservative is selected from the group consisting of m-cresol, phenol, and benzyl alcohol.1 10. The method of claim 108 or 109, wherein the preservative is present in the pharmaceutical composition or formulation in an amount of about 0.1 wt% to about 1 .0 wt%, relative to a total weight of the composition or formulation.-81 -168785213.5

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