Orally deliverable non-naturally occurring melanocortin analogs

Abstract

Provided herein are non-naturally occurring melanocortin analogs. The non-naturally occurring melanocortin analogs of the present technology may be useful in treating, preventing, reducing, or otherwise ameliorating one or more disorders associated with the melanocortin system. In some embodiments, the non-naturally occurring melanocortin analog is administered orally to a subject in need thereof.

Description

ORALLY DELIVERABLE NON-NATURALLY OCCURRINGMELANOCORTIN ANALOGSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 554,944, filed February 16, 2024; U.S. Provisional Patent Application No. 63 / 557,409, filed February 23, 2024; U.S. Provisional Patent Application No. 63 / 572,896, filed April 1 , 2024; U.S. Provisional Patent Application No. 63 / 573,433, filed April 2, 2024; U.S. Provisional Patent Application No. 63 / 632,496, filed April 10, 2024; U.S. Provisional Patent Application No. 63 / 637,285, filed April 22, 2024; U.S. Provisional Patent Application No. 63 / 640,864, filed April 30, 2024; U.S. Provisional Patent Application No. 63 / 647,545, filed May 14, 2024; U.S. Provisional Patent Application No. 63 / 650,368, filed May 21 , 2024; U.S. Provisional Patent Application No. 63 / 654,876, filed May 31 , 2024; U.S. Provisional Patent Application No. 63 / 656,579, field June 5, 2024; U.S. Provisional Patent Application No. 63 / 663,689, filed June 24, 2024; U.S. Provisional Patent Application No. 63 / 675,219, filed July 24, 2024; U.S. Provisional Patent Application No. 63 / 681 ,055, filed August 8, 2024; and of U.S. Provisional Patent Application No. 63 / 692,605, filed September 9, 2024, all of which are incorporated herein by reference in their 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 hereby incorporated by reference in its entirety. The xml copy, created on February 13, 2025, is titled “146316_8023_WO03_SL.xml” and is 363,952 bytes in size.BACKGROUND

[0003] Melanocortins are a group of small peptides that bind to a family of five known melanocortin receptors (MC1 R through MC5R). They are derived from a common precursor protein, pro-opiomelanocortin (POMC), which is expressed in the neurons of the central and peripheral nervous system, and in the pituitary gland. The proteolytic cleavage of POMCresults in a-, [3- and y-melanocortin and adrenocorticotrophic hormone (ACTH), in addition to several other biologically important peptides.

[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. However, in regulating these physiological effects, melanocortin analogs have also been shown to cause certain side effects, such as, for example hypertension and cardiac arrhythmias.

[0005] Despite recent attempts to reduce melanocortin peptide side effects or enhance melanocortin in vivo activity, there still exists a need for highly stable and selective melanocortin analogs in the treatment and prevention of various melanocortin-related diseases.SUMMARY

[0006] Provided herein are non-naturally occurring melanocortin analogs comprising a sequence of Formula (IA(iii)):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA(iii)), wherein:X1is Nle;R1is Asp or Glu;R2is Pro;R3is His;R4is dPhe;R5is Arg;R6is Trp;R7is Lys or Orn;Y1is dVal;Y2is dPro; andthe non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp or Glu at R1and Lys or Orn at R7.

[0007] In some embodiments, the non-naturally occurring melanocortin analog is administered via oral administration.BRIEF DESCRIPTION OF THE DRAWINGS

[0008] FIG 1. illustrates 24-hour plasma concentrations of non-naturally occurring melanocortin analogs of the present technology following oral administration of Compounds Ac-Nle-c[Asp-Pro-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 80; F2) and Ac-Nle- c[Asp-His-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 74; F5) to Cynomolgus Monkeys at 30.0 mg / kg.

[0009] FIGS. 2A-2I illustrates 24-hour plasma concentrations of non-naturally occurring melanocortin analogs of the present technology following PO administration of Compound D (Ac-Nle-c[Glu-Pro-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2(SEQ ID NO: 118; 07)) (FIGS. 2A-2C) or Compound E (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2(SEQ ID NO: 117; O10)) (FIGS. 2D-2I) to Cynomolgus Monkeys at 3 mg / kg, 10.0 mg / kg, 30.0 mg / kg, or 60 mg / kg.

[0010] FIGS. 3A-3F illustrates 24-hour plasma and CSF concentrations of non- naturally occurring melanocortin analogs of the present technology following PO administration of Compounds D (Ac-Nle-c[Glu-Pro-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2 (SEQ ID NO: 118; 07)) (FIGS. 3A-3D) and E (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]- dVal-dPro-NH2(SEQ ID NO: 117; 010)) (FIGS. 3E and 3F) of FIGS. 2A-2I to Rats at 10.0 mg / kg and 30.0 mg / kg.

[0011] FIGS. 4A-4C show changes in daily caloric intake (FIG. 4A), cumulative caloric intake (FIG. 4B), percent change in caloric consumption from baseline through day 6 (FIG. 4C) for diet-induced obese monkeys orally administered 10 mg / kg of 07 (Ac-Nle-c[Glu-Pro- p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 118) or 010 (Ac-Nle-c[Glu-His- p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 117).

[0012] FIGS. 5A-5P show changes in cumulative caloric intake, percent change in caloric consumption from baseline, food intake of a normal diet, food intake of a high fat diet,food intake in calories, and food preference for diet-induced obese monkeys orally administered 07 (Ac-Nle-c[Glu-Pro-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 118), 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 117), 011 (Ac-Nle-c(Glu-Pro-dPhe-Arg-Trp-Orn)-dVal-dPro-NH2; SEQ ID NO: 119), or A07D (Ac-Nle- c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 56).

[0013] FIG. 5Q shows normalized cumulative food consumption in rats subcutaneously administered MC4R selective agonists (A07D, 07, 011 ) (n=5 per group) or MC3R / MC4R co-agonists (010) (n=5) compared to saline controls (n=4). Rats were administered 0.5 mg / kg A07D, 07, 010, or 011 for days 1 -7 and 1 mg / kg for days 8-17.

[0014] FIG. 5R shows an average percent change in caloric consumption from baseline in moneys orally administered 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro- NH2; SEQ ID NO: 117) at a starting dose of 10 mg / kg and increased to 20 mg / kg.

[0015] FIGS. 6A-6K show changes in body weight for diet included obese monkeys orally administered 07 (Ac-Nle-c[Glu-Pro-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 118) or 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 117).

[0016] FIG. 6L shows changes in body weight as a percentage of day 1 in rats administered saline, a melanocortin 4 receptor (MC4R) selective agonist (A07D (Ac-Nle- c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 56), 07 (Ac-Nle-c[Glu-Pro- p(F)dPhe-Arg-Trp-Om]-dVal-dPro-NH2; SEQ ID NO: 118), or O11 (Ac-Nle-c(Glu-Pro-dPhe- Arg-Trp-Orn)-dVal-dPro-NH2; SEQ ID NO: 119)), or a melanocortin 3 receptor (MC3R) / MC4R coagonist ((010)(Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 117)).

[0017] FIG. 6M shows an average percent change in normalized body weight from baseline in moneys orally administered 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]- dVal-dPro-NH2; SEQ ID NO: 117) at a starting dose of 10 mg / kg and increased to 20 mg / kg.

[0018] FIGS. 7A-7N show changes in body composition for the diet-induced obese monkeys of which the caloric intake data is presented in FIGS. 14A-14C.

[0019] FIGS. 8A-8D show blood chemistry levels in diet-induced obese monkeys orally administered 10 mg / kg 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 117) once daily for week 1 , 20mg / kg 010 once daily for week 2, and 15 mg / kg 010 twice daily weeks 3 and 4.

[0020] FIGS. 9A-9C show measurements of systolic blood pressure (FIG. 9A), diastolic blood pressure (FIG. 9B), and heart rate (FIG. 9C) in rats subcutaneously administered 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Om]-dVal-dPro-NH2; SEQ ID NO: 117) at 0.5 mg / kg, 1 mg / kg, or 3 mg / kg.

[0021] FIGS. 10A-1 OD show measurements of diastolic blood pressure (DBP), systolic blood pressure (SBP), heart rate, and heart rate corrected QT interval (QTc) in rats administered setmelanotide at 0.5 mg / kg, 1.0 mg / kg, 3.0 mg / kg, or 6.0 mg / kg, relative to control rats administered saline.

[0022] FIGS. 11A-11 H show measurements of normalized heart rate (FIG. 11 A), QTc (FIG. 11 B), systolic blood pressure (FIG. 11 C), diastolic blood pressure (FIG. 11 D), mean blood pressure (FIG. 11 E), QT interval (FIG. 11 F), QRS internal (FIG. 11 G), and RR internal (FIG. 11 H) in male cynomolgus monkeys orally (PO) administered 010 (Ac-Nle-c[Glu-His- p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 117) at 3, 10, 30, or 60 mg / kg, relative to those subcutaneously (SC) administered setmelanotide at 3 mg / kg or orally administered saline. (N = 3 to 5 animals per group).

[0023] FIG. 12 shows results of 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal- dPro-NH2; SEQ ID NO: 117) screened for binding against a panel of 87 targets at 10pM. X- axis represents the percent inhibition of control at 10 M. (MC1 R: melanocortin 1 receptor; MC4R: melanocortin 4 receptor).

[0024] FIG. 13 shows exemplary adipose tissue staining in diet-induced obese monkeys orally administered 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro- NH2; SEQ ID NO: 117) at 10 mg / kg.

[0025] FIG. 14 shows plasma concentration of 07 after oral administration to cynomolgus monkeys at 10 mg / kg or 30 mg / kg.

[0026] FIG. 15 shows plasma concentration of 010 (Ac-Nle-c[Glu-His-p(F)dPhe-Arg- Trp-Orn]-dVal-dPro-NH2; SEQ ID NO: 117) in monkeys orally administered 3mg / kg, 10 mg / kg, 30 mg / kg, or 60 mg / kg O10.DETAILED DESCRIPTION

[0027] The present technology comprises methods of treating, preventing, or reducing one or more symptoms or conditions associated with metabolic dysfunction in a subject in need thereof using a non-naturally occurring melanocortin analog. In some embodiments, the non-naturally occurring melanocortin analog is administered orally. In some embodiments, the method comprises suppressing appetite in a subject in need thereof using a non-naturally occurring melanocortin analog in accordance with the present technology. In some embodiments, the method comprises promoting fat loss in a subject in need thereof using a non-naturally occurring melanocortin analog in accordance with the present technology. In some embodiments, the method comprises reducing body weight and / or fat mass in a subject in need thereof using a non-naturally occurring melanocortin analog, or a pharmaceutical composition thereof, of the present technology.

[0028] 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. 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.

[0029] 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 orfunctions 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

[0030] 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 present technology belongs. For the purposes of the present technology, the following terms are defined below.

[0031] 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. Byway of example, “an element” means one element or more than one element.

[0032] 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.

[0033] The terms “administering” or “administer” include delivery of therapies (e.g., combination therapies, non-naturally occurring melanocortin analogs (also referred to herein as peptides and synthetic peptides), of the present technology to a subject either by local or systemic administration. Administration may be topical (including ophthalmic and to mucous membranes including vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer), intratracheal, intranasal, epidermal and transdermal, oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration.

[0034] 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 thepharmaceutical composition which elicits the intended effect of an administered therapeutic. This may be any component that drives the pharmacological activity or direct effect in the diagnosis, cure, mitigation, treatment, or prevention of the conditions associated with the present technology, such as but not limited to, reduced appetite and weight loss.

[0035] 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.

[0036] As used herein, a “pharmaceutically acceptable carrier” of the first or the pharmaceutical composition 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. The term “carrier” encompasses any excipient, binder, diluent, filler, salt, buffer, solubilizer, lipid, stabilizer, or other material well known in the art for use in pharmaceutical formulations. The choice of a carrier for use in a composition will depend upon the intended route of administration for the composition. The preparation of pharmaceutically acceptable carriers and formulations containing these materials is described in, e.g., Remington's Pharmaceutical Sciences, 21 st Edition, ed. University of the Sciences in Philadelphia, Lippincott, Williams & Wilkins, Philadelphia Pa., 2005, which is incorporated herein by reference in its entirety). Some examples of physiologically acceptable carriers include antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN® (ICI, Inc.; Bridgewater, N.J.), polyethylene glycol (PEG), and PLURONICS™ (BASF; Florham Park, N.J.). An “excipient” of the first or the pharmaceutical composition refers to an inert substance added to a composition to further facilitate administration of a compound. Examples, without limitation,of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0037] The term “weight loss agent” refers to a therapeutic agent useful for the treatment or prevention of metabolic dysfunction or one or more symptoms associated with metabolic dysfunction in a subject. In some embodiments, such weight loss agents may be effective to treat, reduce, prevent, or otherwise be useful for a subject having a disease or condition that is not metabolic dysfunction, or besides metabolic dysfunction. The present technology is expected to be useful for subjects that may receive, have received, or are receiving one or more doses of a weight loss agent regardless of the underlying disease or condition that the subject has or develops.

[0038] The terms “treat,” “treatment,” and “treating” may also refer to the reduction or inhibition of the progression and / or duration of a disease (e.g., metabolic dysfunction), the reduction or amelioration of the severity of the disease, and / or the amelioration of one or more symptoms thereof resulting from the administration of one or more therapies.

[0039] As used herein, the term “prevent,” “preventing,” or “prevention” means no disorder or disease development if none had occurred, or no further disorder or disease development if there had already been development of the disorder or disease. Also considered is the ability of one to prevent some or all of the symptoms associated with the disorder or disease.

[0040] As used herein, the terms “effective amount” or “therapeutically effective amount ,” refer to that amount of the active ingredient being administered which will relieve to some extent one or more of the symptoms of the disease being treated. The result may be a reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate “effective amount” may differ from one individual to another. An appropriate “effective amount” in any individual case may be determined using techniques, such as a dose escalation study.

[0041] The term “after administration” refers to any duration of time after the non- naturally occurring melanocortin analog or pharmaceutical composition thereof, and / or the weight loss agent has been administered to a subject. “After administration” may also referto the duration of time after one dose has been completed or after more than one dose has been completed, such as two doses, three doses, four doses, and the like. In some embodiments, “after administration” refers to completion of dosing regimen that includes one or more doses. Likewise, the term “prior to administration” refers to any duration of time before the non-naturally occurring melanocortin analog or pharmaceutical composition thereof, and / or the weight loss agent has been administered to a subject. Unless otherwise specified, durations of time encompassed by “after administration” or “prior to administration” may include seconds, minutes, hours, days, weeks, months, and years.

[0042] The term “Body Mass Index” or “BMI” refers to a value derived from an individual’s body weight and height. Specifically, BMI is determined by body weight (kilograms) divided by the square of height (m2) and is expressed in units of “kg / m2”. "Normal" BMI ranges are known to a person of ordinary skill in the art and consider factors such as patient sex, age, height, race, and body type. Typically, a normal BMI range is about 18.5 kg / m2 to about 25 kg / m2.

[0043] The terms “subject” and “patient” refer to anyone being evaluated for disease or condition or being administered a therapeutic or pharmaceutical composition. This includes people without diagnosed or confirmed disease or condition. This also includes people with diagnosed or confirmed disease or condition, such as metabolic dysfunction.

[0044] The term “control subject,” as used herein, refers to any subject used as a basis for comparison to the subject (e.g., test subject). A control subject includes, but is not limited to, any subject who has not been administered the therapeutic or pharmaceutical composition (e.g., the non-naturally occurring melanocortin analog, a therapeutically effective amount of the non-naturally occurring melanocortin analog or a pharmaceutical composition thereof) or administered a placebo.

[0045] “Melanocortin analogs,” “non-naturally occurring melanocortin analogs,” “melanocortin peptides,” “melanocortin receptor 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 naturally occurring non-naturally occurring melanocortin analogs (i.e.,“synthetic peptides” or “synthetic analogs”) and truncated and / or modified versions of melanocortin full-length protein or peptides. For example, the full-length proopiomelanocortin 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 Lawry, 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. The binding to the melanocortin receptor is inhibitory (antagonist). In addition to peptides, the non-naturally occurring 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. “Non-naturally occurring melanocortin analogs” may be structurally similar and / or functionally similar to biological melanocortin proteins in their ability to bind melanocortin receptors. Further, the non- naturally occurring 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.

[0046] 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 and their analogs are the pharmacophore of melanocortin for the regulated physiological effect. Therefore, non-naturally occurring melanocortin pharmacophore analogs may 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.

[0047] A “melanocortin receptor agonist” or “melanocortin agonist” is a naturally occurring substance or manufactured drug substance or composition that may interact with a melanocortin receptor and initiate a pharmacological response characteristic of the melanocortin receptor.

[0048] The terms “bind,” “binding,” “complex,” and “complexing,” refer to all types of physical and chemical binding, reactions, complexing, attraction, chelating, and the like.

[0049] The “peptides” of the present technology may 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.

[0050] 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.

[0051] 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.

[0052] “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 presenttechnology 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 amino 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.

[0053] 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 of the present technology 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.

[0054] 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.

[0055] In the peptides of the present technology, conventional amino acid residues have their conventional meaning as given in Chapter 2400, of the Manual of PatentExamining 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; “He” 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 may be used. Thus, for example, “L-Phe” or “IPhe” is L-phenylalanine; “D-Phe” or “dPhe” is D- phenylalanine; 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.

[0056] 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 a-carbon). Other types of amino acids exist when the amino group is attached to a different carbon atom. For example, beta (P)-amino acids, the carbon atom to which the amino group is attached is separated from the carboxylate group by one carbon atom, Cp.

[0057] When [3-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.

[0058] 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

[0059] 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” refers to a cyclic structure, which is also shown in the literature as “c” or referred to as a “lactam.” “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.

[0060] Additional abbreviations are used as follows: Pen is Penicillamine; Tic is tetrahydroquinoline-3-carboxylic acid; dBip is D-biphenylalanine; Nal(1 ’) is T- naphthylalanine; Nal(2’) is 2’-naphthylalanine; Orn is ornithine; Bip is biphenylalanine; and p(CI)Phe is para-chloro-phenylalanine (I - iodo, F - fluoro).

[0061] A peptide or aliphatic moiety 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.

[0062] An “amine” includes compounds that contain an amine group ( — NH2).

[0063] 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.

[0064] Amino acids, including stereoisomers and modifications of naturally occurring amino acids, 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 (peptide mimetics), and the like, including all of the foregoing, are sometimes referred to herein as “residues.”

[0065] “Substantial degradation” refers to the degradation of the N-terminal extension, the C-terminal extension, both N- and C-terminal degradation or degradation to other regions of the melanocortin peptide by physiological enzymes and other factors, in such a manner or to a degree that side effects appear. According to one aspect, a melanocortin analog having a C-terminal extension that resists substantial degradation is one where no more than 50% of the administered peptide causes side effects and / or displays a low half-life. In some aspects, no more than 25% of the administered peptide causes side effects and / or displays a low half-life. More preferably, in some aspects, less than 10% of the administered peptide causes side effects and / or displays a low half-life, as compared to a melanocortin analog that lacks a C-terminal extension.

[0066] As used herein, a “composition” refers to a mixture of the active ingredient with other chemical components.

[0067] The disclosure of all publications, patents, and published patent applications listed herein are hereby incorporated by reference in their entireties, including but not limited to U.S. Patent Nos. 8,541 ,545 and 9,534,018.Non-naturally Occurring Melanocortin Analogs

[0068] The non-naturally occurring melanocortin analogs of the present technology may comprise a non-naturally occurring melanocortin analog or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof. Non-naturally occurring melanocortin analogs of the present technology may be selective for the melanocortin 3 receptor (MC3R) over other melanocortin receptors, i.e., the melanocortin 1 receptor (MC1 R), the melanocortin 2 receptor (MC2R), the melanocortin 1 receptor (MC4R), and the melanocortin 5 receptor (MC5R). Some of the non-naturally occurring melanocortin analogs may bind only the MC3R.

[0069] Alternatively, the non-naturally occurring melanocortin analogs of the present technology may be selective for the MC1 R and / or MC5R over other melanocortin receptors, i.e., the MC2R, MC3R, and MC4R. Some of the non-naturally occurring melanocortin analogs may bind only the MC1 R or the MC5R. Alternatively, some of the non-naturally occurring melanocortin analogs may bind the MC1 R with greater affinity than the MC5R, whereas other melanocortin analogs may bind the MC5R with greater affinity than the MC1 R. Certain melanocortin analogs may bind the MC1 R with the same or generally similar affinity as the MC5R.

[0070] The non-naturally occurring melanocortin analogs of the present technology may be full agonists or full antagonists for one or more melanocortin receptors. A full agonist may comprise a non-naturally occurring melanocortin analog having a maximum effect (Emax) agonist value of greater than or equal to 85%. Similarly, a full antagonist may comprise a non-naturally occurring melanocortin analog having an Emax antagonist value of greater than or equal to 85%.

[0071] The non-naturally occurring melanocortin analogs of the present technology may be partial agonists or partial antagonists A partial agonist may comprise a non-naturally occurring melanocortin analog having a maximum effect Emax agonist value of less than85%. Similarly, a partial antagonist may comprise a non-naturally occurring melanocortin analog having an Emax antagonist value of less than 85%.

[0072] If a non-naturally occurring melanocortin analog’s Emax agonist value is greater than it’s Emax antagonist value, then the non-naturally occurring melanocortin analog may be classified as an agonist (e.g., a full agonist or a partial agonist).

[0073] If a non-naturally occurring melanocortin analog’s Emax antagonist value is greater than it’s Emax agonist value, then the non-naturally occurring melanocortin analog may be classified as an antagonist (e.g., a full antagonist or a partial antagonist).

[0074] The non-naturally occurring melanocortin analogs of the present technology may be one or more of (i) a full MC3R antagonist having no MC4R activity; (ii) a partial MC3R antagonist having no MC4R activity; (iii) a full MC3R agonist having no MC4R activity, (iv) a partial MC3R agonist having no MC4R activity; (v) a full MC1 R agonist and a partial MC5R agonist; (vi) a full MC1 R agonist and a full MC5R agonist; (vii) a full MC5R agonist having no MC1 R activity; (viii) a partial MC5R agonist having no MC1 R activity); (ix) a full MC1 R agonist having no MC5R activity; (x) a partial MC1 R agonist having no MC5R activity; and (xi) a partial MC1 R agonist and a full MC5R agonist.

[0075] The non-naturally occurring melanocortin analogs of the present technology may avoid cardiac activation typically seen in conventional melanocortin peptide and small molecule agonists. For example, a subject may maintain a stable heart rate, systolic blood pressure, and / or diastolic blood pressure following administration of a melanocortin analog of the present technology.

[0076] The non-naturally occurring melanocortin analogs in accordance with the present technology may have certain structural features that impart specific properties on the analogs, such as, for example, degradation resistance, enhanced epithelial, gastrointestinal, and / or blood brain barrier transport, and binding affinity for the melanocortin 4 receptor and / or melanocortin 3 receptor. Accordingly, in some embodiments, the non- naturally occurring melanocortin analogs have one or more beta hairpin (P-hairpin) and / or beta turn (|3-turn) structures. The presence of amino acids that are structurally rigid, such as, for example, Pro and dPro, may lead to formation of p-hairpin and / or p-turn structures inthe non-naturally occurring melanocortin analog. Additionally, dMet and disulfide bridges (e.g., cyclization via disulfide bond), as well as Pro-Gly and Gly-Gly moieties may induce and / or stabilize beta-turn structures of the non-naturally occurring melanocortin analogs. In general, cyclization may stabilize beta-turns, and D-amino acids may induce and / or stabilize beta-turns. Further, in some embodiments, melanocortin analogs include D-valine-D-proline (dVal-dPro) chain as their C-terminus, which may provide enhanced transport and resistance to degradation.

[0077] In some embodiments, the non-naturally occurring melanocortin analog has activity on one or more of MC1 R, MC3R, and MC5R. In some embodiments, the non- naturally occurring melanocortin analog comprises a sequence of Formula (I):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(I), wherein:X1is absent or norleucine (Nle);R1is selected from the group consisting of D-penicillamine (dPen), Nle, arginine (Arg), D-arginine (dArg), aspartic acid (Asp), and glutamic acid (Glu);R2is selected from the group consisting of Asp, Arg, tryptophan (Trp), proline (Pro), histidine (His), glycine (Gly), penicillamine (Pen), and dPen;R3is absent or is selected from the group consisting of D-isoleucine (dlle), His, Pro, Arg, D-valine (dVal), Gly, D-tryptophan (dTrp), alanine (Ala), D-alanine (dAla), leucine (Leu), D-leucine (dLeu), D-phenylalanine (dPhe), D-glutamine (dGIn), and D-biphenylalanine (dBip);R4is selected from the group consisting of dPhe, dBip, 2’-D-naphthylalanine (dNal(2’)), para-chloro-D-phenylalanine (p(CI)dPhe), para-iodo-D-phenylalanine (p(l)dPhe), and para-fluoro-D-phenylalanine (p(F)dPhe);R5is selected from the group consisting of Arg, Pro, and His;R6is selected from the group consisting of Trp, phenylalanine (Phe), dPhe, T-D- naphthylalanine (dNal(1 ’)), and tetrahydro-isoquinoline-3-carboxylic acid (Tic);R7is selected from the group consisting of lysine (Lys), cysteine (Cys), dPen, and ornithine (Orn);Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R1or R2and R7when R1or R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; a lactam bridge between R1or R2and R7when R1or R2is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn, provided that: when R4is dPhe, then (i) R3is absent, dlle, dVal, dLeu, or dAla; or (ii) R3is His and R5is Pro or His; or (iii) R3is Pro and R1is Nle or Asp, wherein when R1is Nle, then R5is Pro or R6is dNal(1 ’), and when R1is Asp, then (i) R2is Pro or (ii) R2is Trp and either R5is His or R6is selected from Phe, dPhe, and dNal(1 ’); when R4is p(F)dPhe, then R3is selected from dPhe, dBip, and dTrp; when R4is p(CI)dPhe, then R3is selected from dBip, dGIn, and dTrp; when R4is p(l)dPhe, then R6is Tic; and when R4is dNal(2’), then R3is not Pro and the non-naturally occurring melanocortin analog is cyclized through a disulfide bond between dPen at R1or R2and dPen at R7.

[0078] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I), wherein:X1is absent or norleucine (Nle);R1is selected from the group consisting of D-penicillamine (dPen), Nle, arginine (Arg), D-arginine (dArg), aspartic acid (Asp), and glutamic acid (Glu);R2is selected from the group consisting of Asp, Arg, tryptophan (Trp), proline (Pro), histidine (His), glycine (Gly), penicillamine (Pen), and dPen;R3is absent or is selected from the group consisting of D-isoleucine (dlle), His, Pro, Arg, D-valine (dVal), Gly, D-tryptophan (dTrp), alanine (Ala), D-alanine (dAla), leucine (Leu), D-leucine (dLeu), D-phenylalanine (dPhe), D-glutamine (dGIn), and D-biphenylalanine (dBip);R4is selected from the group consisting of dPhe, dBip, 2’-D-naphthylalanine (dNal(2’)), para-chloro-D-phenylalanine (p(CI)dPhe), para-iodo-D-phenylalanine (p(l)dPhe), and para-fluoro-D-phenylalanine (p(F)dPhe);R5is selected from the group consisting of Arg, Pro, and His;R6is selected from the group consisting of Trp, phenylalanine (Phe), dPhe, T-D- naphthylalanine (dNal(T)), and tetrahydro-isoquinoline-3-carboxylic acid (Tic);R7is selected from the group consisting of lysine (Lys), cysteine (Cys), dPen, and ornithine (Orn);Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R1or R2and R7when R1or R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; a lactam bridge between R1or R2and R7when R1or R2is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn, provided that the non-naturally occurring melanocortin analog does not comprise a sequence selected from the group consisting of:Ac-Nle-c[dPen-Pro-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 38);Ac-dArg-c[dPen-Pro-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 39);Ac-Arg-c[dPen-Pro-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 40);Ac-Nle-c[dPen-Pro-dPhe-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 41 );Ac-Nle-c[Asp-Ala-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 42);Ac-Nle-c[Asp-dBip-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 43);Ac-Nle-c[Asp-dGln-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 44);Ac-Nle-c[Asp-dPhe-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 45);Ac-Nle-c[Asp-dTrp-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 46);Ac-Nle-c[Asp-Trp-Ala-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 47);Ac-Nle-c[Asp-Trp-Leu-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 48);Ac-Nle-c[Asp-Trp-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 49);Ac-Nle-c[Asp-Leu-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 50);Ac-Nle-c[Asp-Phe-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 51 );Ac-dPhe-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 52);Ac-Nle-c[Asp-Pro-dPhe-Arg-Nal(1’)-Lys]-dVal-dPro-NH2(SEQ ID NO: 53);Ac-Nle-c[Asp-Pro-dPhe-Arg-Phe-Lys]-dVal-dPro-NH2(SEQ ID NO: 54);Ac-Nle-c[Asp-Trp-Pro-dPhe-Arg-Nal(1')-Lys]-dVal-dPro-NH2(SEQ ID NO: 55);Ac-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 56);Ac-Ala-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 57);Ac-dArg-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 58);Ac-Arg-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 59);Ac-Nle-c[Asp-His-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 60);Ac-Nle-c[Asp-Pro-dPhe-His-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 61 );Ac-Nle-c[Asp-Trp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 62);Ac-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-OH (SEQ ID NO: 63);Ac-Nle-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 64);Ac-Nle-c[Asp-His-dPhe-Arg-Trp-Lys]-d al-dPro-NH2 (SEQ ID NO: 65);Ac-Nle-c[Asp-Pro-His-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 66);Ac-Nle-c[Asp-dGln-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 67);Ac-Nle-c[Asp-Trp-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 68);Ac-Ala-c[Asp-His-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 69);Ac-dArg-c[Asp-His-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 70);Ac-Arg-c[Asp-His-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 71 );Ac-Nle-Nle-c[Asp-His-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 72);Ac-Nle-c[Asp-His-p(F)dPhe-His-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 73);Ac-Nle-c[Asp-His-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 74);Ac-Ala-c[Asp-Pro-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 75);Ac-dArg-c[Asp-Pro-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 76);Ac-Arg-c[Asp-Pro-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 77);Ac-Nle-Nle-c[Asp-Pro-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 78);Ac-Nle-c[Asp-Pro-p(F)dPhe-His-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 79);Ac-Nle-c[Asp-Pro-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 80);Ac-Nle-c[Asp-dPhe-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 81 );Ac-Nle-c[Asp-Trp-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 82);Ac-Nle-c[Asp-Pro-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 83);Ac-dArg-c[Asp-His-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 84);Ac-Arg-c[Asp-His-p(CI)dPhe-Arg-Trp-Lys]-d al-dPro-NH2 (SEQ ID NO: 85);Ac-Nle-Nle-c[Asp-His-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2 (SEQ ID NO: 86);Ac-Nle-c[Asp-His-p(CI)dPhe-His-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 87);Ac-Nle-c[Asp-His-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 88);Ac-Nle-c[Asp-Pro-p(l)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 89);Ac-Nle-c[Asp-His-p(l)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 90);Ac-dArg-c[Asp-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 91 );Ac-Nle-c[Asp-Gly-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 92);Ac-Nle-c[Asp-Pro-His-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 93);Ac-Nle-c[Asp-Trp-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 94);Ac-Nle-c[Asp-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 95);Ac-Nle-c[Asp-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 96);Ac-Nle-c[Asp-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro-OH (SEQ ID NO: 97);Ac-Nle-c[Asp-Pro-dNal(2')-His-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 98);Ac-Nle-c[Pen-Pro-dNal(2')-Arg-Trp-Cys]-dVal-dPro-NH2(SEQ ID NO: 99);Ac-Nle-c[Asp-Pro-dNal(2')-Arg-Phe-Lys]-dVal-dPro-NH2(SEQ ID NO: 100);Ac-Nle-c[Glu-Pro-dNal(2')-Arg-Trp-Orn]-dVal-dPro-NH2(SEQ ID NO: 101 );Ac-Nle-c[Asp-Pro-dNal(2')-Arg-dPhe-Lys]-dVal-dPro-NH2(SEQ ID NO: 102);Ac-Nle-c[Asp-Ala-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 103);Ac-Nle-c[Asp-dAla-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 104);Ac-Nle-c[Asp-Gly-Gly-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 105);Ac-Nle-c[Asp-dVal-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 106);Ac-Nle-c[Asp-Trp-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 107);Ac-Nle-Nle-c[Asp-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 108);Ac-Nle-c[Asp-Pro-dNal(2')-Arg-dNal(1 ’)-Lys]-dVal-dPro-NH2(SEQ ID NO: 109);Ac-Nle-c[Asp-Leu-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 110);Ac-Nle-c[Asp-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro (SEQ ID NO: 111);Ac-Nle-c[Asp-Pro-Gly-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 112);Ac-Nle-c[Asp-dLeu-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 113);Ac-Arg-c[Asp-dAla-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 114);Ac-dArg-c[Asp-dAla-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 115); andAc-Nle-c[Asp-Pro-Pro-dNal(2')-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 116).

[0079] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I), wherein R4is dPhe. Accordingly, in some embodiments, the sequence of Formula (I) is a sequence of Formula (IA):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA), wherein:X1is absent or Nle;R1is selected from the group consisting of Nle, Arg, dArg, Asp, and Glu;R2is selected from the group consisting of Asp, Trp, Pro, Pen, and dPen;R3is absent or is selected from the group consisting of Pro, His, dVal, dLeu, dAla, and dlle;R4is dPhe;R5is selected from the group consisting of Arg, Pro, and His;R6is selected from the group consisting of Trp, Phe, dPhe, and dNal(1 ’);R7is selected from the group consisting of Lys, Cys, dPen, and Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R2and R7when R2and R7are dPen;a disulfide bond between R2and R7when R2is Pen and R7is Cys; a lactam bridge between R1or R2and R7when R1or R2is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn.

[0080] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA), wherein:X1is absent or Nle;R1is selected from the group consisting of Nle, Arg, dArg, Asp, and Glu;R2is selected from the group consisting of Asp, Trp, Pro, Pen, and dPen;R3is absent or is selected from the group consisting of Pro, His, dVal, dLeu, dAla, and dlle;R4is dPhe;R5is selected from the group consisting of Arg, Pro, and His;R6is selected from the group consisting of Trp, Phe, dPhe, and dNal(1 ’);R7is selected from the group consisting of Lys, Cys, dPen, and Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R2and R7when R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; a lactam bridge between R1or R2and R7when R1or R2is Asp and R7is Lys; anda lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn, provided that: when R3is His, then R5is Pro or His; and when R3is Pro, then R1is Nle or Asp, wherein when R1is Nle, then R5is Pro or R6is dNal(1 ’), and when R1is Asp, then (i) R2is Pro or (ii) R2is Trp and either R5is His or R6is selected from Phe, dPhe, and dNal(1’).

[0081] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA), wherein:X1is absent or Nle;R1is selected from the group consisting of Nle, Arg, dArg, Asp, and Glu;R2is selected from the group consisting of Asp, Trp, Pro, Pen, and dPen;R3is absent or is selected from the group consisting of Pro, His, dVal, dLeu, dAla, and dlle;R4is dPhe;R5is selected from the group consisting of Arg, Pro, and His;R6is selected from the group consisting of Trp, Phe, dPhe, and dNal(1 ’);R7is selected from the group consisting of Lys, Cys, dPen, and Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R2and R7when R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; a lactam bridge between R1or R2and R7when R1or R2is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn,provided that the non-naturally occurring melanocortin analog does not comprise a sequence selected from the group consisting of:Ac-Nle-c[dPen-Pro-dPhe-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 41 );Ac-Nle-c[Asp-Ala-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 42);Ac-Nle-c[Asp-dBip-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 43);Ac-Nle-c[Asp-dGln-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 44);Ac-Nle-c[Asp-dPhe-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 45);Ac-Nle-c[Asp-dTrp-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 46);Ac-Nle-c[Asp-Trp-Ala-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 47);Ac-Nle-c[Asp-Trp-Leu-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 48);Ac-Nle-c[Asp-Trp-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 49);Ac-Nle-c[Asp-Leu-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 50);Ac-Nle-c[Asp-Phe-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 51 );Ac-dPhe-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 52);Ac-Nle-c[Asp-Pro-dPhe-Arg-Nal(1’)-Lys]-dVal-dPro-NH2(SEQ ID NO: 53);Ac-Nle-c[Asp-Pro-dPhe-Arg-Phe-Lys]-dVal-dPro-NH2(SEQ ID NO: 54);Ac-Nle-c[Asp-Trp-Pro-dPhe-Arg-Nal(1 ')-Lys]-dVal-dPro-NH2(SEQ ID NO: 55);Ac-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 56);Ac-Ala-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 57);Ac-dArg-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 58);Ac-Arg-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 59);Ac-Nle-c[Asp-His-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 60);Ac-Nle-c[Asp-Pro-dPhe-His-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 61 );Ac-Nle-c[Asp-Trp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 62); andAc-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-OH (SEQ ID NO: 63).

[0082] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I) or (IA), wherein R4is dPhe and X1is absent. Accordingly, in some embodiments, the sequence of Formula (I) of (IA) is a sequence of Formula (IA(i)):R1-R2-R3-R4-R5-R6-R7-Y1-Y2( I A( i)), wherein:R1is selected from the group consisting of Nle, Arg, and dArg;R2is selected from the group consisting of Asp, Pen, and dPen;R3is absent or is selected from the group consisting of Pro, His, dVal, dAla, and dlle;R4is dPhe;R5is Arg or Pro;R6is Trp or dNal(T);R7is selected from the group consisting of Lys, Cys, or dPen;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R2and R7when R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; and a lactam bridge between R2and R7when R2is Asp and R7is Lys.

[0083] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(i)):R1-R2-R3-R4-R5-R6-R7-Y1-Y2( I A( i)), wherein:R1is selected from the group consisting of Nle, Arg, and dArg;R2is selected from the group consisting of Asp, Pen, and dPen;R3is absent or is selected from the group consisting of Pro, His, dVal, dAla, and dlle;R4is dPhe;R5is Arg or Pro;R6is Trp or dNal(T);R7is selected from the group consisting of Lys, Cys, or dPen;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R2and R7when R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; and a lactam bridge between R2and R7when R2is Asp and R7is Lys, provided that: when R3is His, then R5is Pro; and when R3is Pro, then R5is Pro or R6is dNal(1 ’)•

[0084] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(i)), wherein:R1is selected from the group consisting of Nle, Arg, and dArg;R2is selected from the group consisting of Asp, Pen, and dPen;R3is absent or is selected from the group consisting of Pro, His, dVal, dAla, and dlle;R4is dPhe;R5is Arg or Pro;R6is Trp or dNal(T);R7is selected from the group consisting of Lys, Cys, or dPen;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a disulfide bond between R2and R7when R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; and a lactam bridge between R2and R7when R2is Asp and R7is Lys, provided that the non-naturally occurring melanocortin analog does not comprise a sequence selected from the group consisting of:Ac-Nle-c[dPen-Pro-dPhe-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 41 );Ac-dPhe-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 52);Ac-Nle-c[Asp-Pro-dPhe-Arg-Nal(1’)-Lys]-dVal-dPro-NH2(SEQ ID NO: 53);Ac-Nle-c[Asp-Pro-dPhe-Arg-Phe-Lys]-dVal-dPro-NH2(SEQ ID NO: 54);Ac-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 56);Ac-Ala-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 57);Ac-dArg-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 58);Ac-Arg-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 59);Ac-Nle-c[Asp-Pro-dPhe-His-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 61 ); andAc-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-OH (SEQ ID NO: 63).

[0085] Alternatively, in some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I) or (IA), wherein R4is dPhe and X1is Nle. Accordingly, in some embodiments, the sequence of Formula (I) of (IA) is a sequence of Formula (IA(ii)):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA(ii)), wherein:X1is Nle;R1is Asp or Glu;R2is Trp or Pro;R3is selected from the group consisting of Pro, His, dVal, and dLeu;R4is dPhe;R5is Arg or His;R6is selected from the group consisting of Trp, Phe, dPhe, and dNal(1 ’);R7is Lys or Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a lactam bridge between R1and R7when R1is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn.

[0086] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(ii)):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA(ii)), wherein:X1is Nle;R1is Asp or Glu;R2is Trp or Pro;R3is selected from the group consisting of Pro, His, dVal, and dLeu;R4is dPhe;R5is Arg or His;R6is selected from the group consisting of Trp, Phe, dPhe, and dNal(1 ’);R7is Lys or Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of: a lactam bridge between R1and R7when R1is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn, provided that: when R3is His, then R5is His; and when R3is Pro, then (i) R2is Pro or (ii) R2is Trp and either R5is His or R6is selected from Phe, dPhe, and dNal(T).

[0087] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(ii)), wherein:X1is Nle;R1is Asp or Glu;R2is Trp or Pro;R3is selected from the group consisting of Pro, His, dVal, and dLeu;R4is dPhe;R5is Arg or His;R6is selected from the group consisting of Trp, Phe, dPhe, and dNal(1 ’);R7is Lys or Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a moiety selected from the group consisting of:a lactam bridge between R1and R7when R1is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn, provided that the non-naturally occurring melanocortin analog does not comprise a sequence of:Ac-Nle-c[Asp-Trp-Pro-dPhe-Arg-Nal(T)-Lys]-dVal-dPro-NH2 (SEQ ID NO: 55); orAc-Nle-c[Asp-Trp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 62).

[0088] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I), (IA) or (IA(ii)), wherein R4is dPhe, X1is Nle, and R3is His. Accordingly, in some embodiments, the sequence of Formula (I) of (IA) is a sequence of Formula (IA(iii)):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA(iii)), wherein:X1is Nle;R1is Asp or Glu;R2is Pro;R3is His;R4is dPhe;R5is Arg;R6is Trp;R7is Lys or Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp or Glu at R1and Lys or Orn at R7.

[0089] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I), wherein R4is p(CI)dPhe or p(F)dPhe. Accordingly, in some embodiments, the sequence of Formula (I) is a sequence of Formula (IB):R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IB), wherein:R1is Nle;R2is Asp;R3is absent or is selected from the group consisting of dBip, dGIn, dTrp, and dPhe;R4is p(CI)dPhe or p(F)dPhe;R5is Arg;R6is Trp;R7is Lys;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp at R2and Lys at R7.

[0090] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IB):R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IB), wherein:R1is Nle;R2is Asp;R3is absent or is selected from the group consisting of dBip, dGIn, dTrp, and dPhe;R4is p(CI)dPhe or p(F)dPhe;R5is Arg;R6is Trp;R7is Lys;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp at R2and Lys at R7, provided that: when R4is p(F)dPhe, then R3is selected from dPhe, dBip, and dTrp; and when R4is p(CI)dPhe, then R3is selected from dBip, dGIn, and dTrp.

[0091] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IB), wherein:R1is Nle;R2is Asp;R3is absent or is selected from the group consisting of dBip, dGIn, dTrp, and dPhe;R4is p(CI)dPhe or p(F)dPhe;R5is Arg;R6is Trp;R7is Lys;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp at R2and Lys at R7, provided that the non-naturally occurring melanocortin analog does not comprise a sequence selected from the group consisting of:Ac-Nle-c[Asp-dGln-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 67); orAc-Nle-c[Asp-dPhe-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 81 ).

[0092] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I), wherein R4is d B ip or p(l)dPhe. In such embodiments, R6is Tic. Accordingly, in some embodiments, the sequence of Formula (I) is a sequence of Formula (IC):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2( IC), wherein:X1is Nle;R1is Asp;R2is selected from the group consisting of His, Pro, and Arg;R3is Arg or Pro;R4is p(l)dPhe or dBip;R5is Arg;R6is Tic;R7is Lys;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp at R1and Lys at R7.

[0093] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IC):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2( IC), wherein:X1is Nle;R1is Asp;R2is selected from the group consisting of His, Pro, and Arg;R3is Arg or Pro;R4is p(l)dPhe or dBip;R5is Arg;R6is Tic;R7is Lys;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp at R1and Lys at R7.

[0094] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I), wherein R4is dNal(2‘). Accordingly, in some embodiments, the sequence of Formula (I) is a sequence of Formula (ID):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(ID), wherein:X1is absent or Nle;R1is Nle or dPen;R2is dPen or Gly;R3is selected from the group consisting of Gly, Ala, Leu, dLeu, and dVal;R4is dNal(2’);R5is Arg;R6is Trp;R7is dPen;Y1is dVal;Y2is dPro; andthe non-naturally occurring melanocortin analog is cyclized through a disulfide bond between dPen at R1or R2and dPen at R7.

[0095] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (ID):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(ID), wherein:X1is absent or Nle;R1is Nle or dPen;R2is dPen or Gly;R3is selected from the group consisting of Gly, Ala, Leu, dLeu, and dVal;R4is dNal(2’);R5is Arg;R6is Trp;R7is dPen;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a disulfide bond between dPen at R1or R2and dPen at R7.

[0096] In some embodiments, the non-naturally occurring melanocortin analog has one or more beta hairpin (P-hairpin) and / or beta turn (P-turn) structures. In some embodiments, R3of the sequence according to any one of Formulae (l)-(ID), which may be D-proline, L- proline, hydroxyproline, D-hydroxyproline, D-alanine, D-methionine, D-valine, prolylglycine (Pro-Gly), glycine, transPro(guan), cisPro(guan), provides the p-hairpin and / or p-turn structures of the non-naturally occurring melanocortin analog. In some embodiments, the disulfide bond of the sequence according to any one of Formulae (l)-(ID), if present, provides the p-hairpin and / or p-tum structures of the non-naturally occurring melanocortin analog.

[0097] As will be appreciated by the skilled artisan, non-naturally occurring melanocortin analogs comprising a sequence of any one of Formulae (l)-(ID), have an N- terminus and a C-terminus. The melanocortin analogs of the present technology are written beginning with the N-terminus at the left-most amino acid residue and ending with the C- terminus at the right most residue. Accordingly, the N-terminus of a non-naturally melanocortin analog comprising a sequence of any one of Formulae (l)-(ID) may be at any of X1, X2, X3, and R1. Analogously, the C-terminus of a non-naturally occurring melanocortin analog comprising a sequence of any one of Formulae (l)-(ID) may be at any of R7, R8, Y1, Y2, Y3, Y4, Y5, Y6, and Y7.

[0098] In some embodiments, the N-terminus of the non-naturally occurring melanocortin analog is modified by an acyl group. In some embodiments, the acyl group is acetyl group

[0099] In some embodiments, the N-terminus of the non-naturally occurring melanocortin analog is not modified.

[0100] As discussed above, Y1Y2Y3Y4Y5Y6Y7represents a C-terminus of the non- naturally occurring melanocortin analog. In some embodiments, Y1-Y7are absent. In some embodiments, Y1is present and Y2-Y7are absent. In some embodiments, Y1and Y2are present and Y3-Y7are absent. In some embodiments, Y1-Y3are present and Y4-Y7are absent. In some embodiments, Y1-Y4are present and Y5-Y7are absent. In some embodiments, Y1-Y5are present and Y6-Y7are absent. In some embodiments, Y1-Y6are present and Y7is absent. In some embodiments, Y1-Y7are present.

[0101] In some embodiments, the C-terminus of the non-naturally occurring melanocortin analog is modified by an amide groupthe sequence of any one of Formulae (l)-(ID), a non-naturally occurring melanocortin analog with a C- terminus modified by an amide may be represented by a terminal -NH2.

[0102] In some embodiments, the C-terminus of the non-naturally occurring melanocortin analog is not modified. In the sequence of any one of Formulae (l)-(ID), a non- naturally occurring melanocortin analog with an unmodified C-terminus may be represented by -OH.

[0103] Non-naturally occurring melanocortin analogs comprising a sequence of any one of Formulae (l)-(ID) are cyclized. For example, the non-naturally occurring melanocortin analog may be cyclized through a moiety selected from the group consisting of: a disulfide bond between R1or R2and R7when R1or R2and R7are dPen; a disulfide bond between R2and R7when R2is Pen and R7is Cys; a lactam bridge between R1or R2and R7when R1or R2is Asp and R7is Lys; and a lactam bridge between R1and R7when R1is Glu, R3is His, and R7is Orn.

[0104] In some embodiments of the sequence of any one of Formulae (l)-(ID), R4is dPhe. In further embodiments, the non-naturally occurring melanocortin analog is cyclized through a lactam bond between R2and R7. In still further embodiments, R1is Nle, R3is selected from Pro, His, dlle, and dVal, R5is Arg or Pro, and R6is Trp or dNal(1 ’)• In some embodiments, the sequence of any one of Formulae (l)-(ID) is selected from the group consisting of:Ac-Nle-c[Asp-His-dPhe-Pro-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 3);Ac-Nle-c[Asp-Pro-dPhe-Pro-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 4);Ac-Nle-c[Asp-dVal-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 14);Ac-Nle-c[Asp-Pro-dPhe-Arg-dNal(1’)-Lys]-dVal-dPro-NH2(SEQ ID NO: 15); andAc-Nle-c[Asp-dlle-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 36), wherein c represents cyclization through R2and R7via a lactam bond.

[0105] In other further embodiments, R1is Arg or dArg, R3is dAla, R5is Arg, and R6is Trp. In some embodiments, the sequence of any one of Formulae (l)-(ID) is: Ac-Arg-c[Asp-dAla-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 31 ); orAc-dArg-c[Asp-dAla-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 32), wherein c represents cyclization through R2and R7via a lactam bond.

[0106] Alternatively, in some embodiments of the sequence of any one of Formulae (I)- (ID), R4is dPhe and the non-naturally occurring melanocortin analog is cyclized through a lactam bond between R1and R7. In further embodiments, R2is Trp and R3is selected from Pro, dLeu and dVal. In some embodiments, the sequence of any one of Formulae (l)-(ID) is selected from the group consisting of:Ac-Nle-c[Asp-Trp-dLeu-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 19);Ac-Nle-c[Asp-Trp-dVal-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 20);Ac-Nle-c[Asp-Trp-Pro-dPhe-His-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 21 );Ac-Nle-c[Asp-Trp-Pro-dPhe-Arg-dNal(1')-Lys]-dVal-dPro-NH2(SEQ ID NO: 22);Ac-Nle-c[Asp-Trp-Pro-dPhe-Arg-Phe-Lys]-dVal-dPro-NH2(SEQ ID NO: 23); andAc-Nle-c[Asp-Trp-Pro-dPhe-Arg-dPhe-Lys]-dVal-dPro-NH2(SEQ ID NO: 24), wherein c represents cyclization through R1and R7via a lactam bond.

[0107] In other further embodiments, R2is Pro and R3is Pro or His. In some embodiments, the sequence of any one of Formulae (l)-(ID) is selected from the group consisting of:Ac-Nle-c[Asp-Pro-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 2);Ac-Nle-c[Asp-Pro-His-dPhe-His-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 35); andAc-Nle-c[Glu-Pro-His-dPhe-His-Trp-Orn]-dVal-dPro-NH2(SEQ ID NO: 37), wherein c represents cyclization through R1and R7via a lactam bond.

[0108] In some embodiments of the sequence of any one of Formulae (l)-(ID), R4is dPhe and the non-naturally occurring melanocortin analog is cyclized through a disulfide bond between R2and R7. In further embodiments, R3is absent. In such embodiments, the sequence of any one of Formulae (l)-(ID) is:Ac-Nle-c[Pen-dPhe-Arg-Trp-Cys]-dVal-dPro-NH2(SEQ ID NO: 33); orAc-Nle-c[dPen-dPhe-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 34), represents cyclization through R2and R7via a disulfide bond.

[0109] In some embodiments of the sequence of any one of Formulae (l)-(ID), R4is p(F)dPhe. In further embodiments, the non-naturally occurring melanocortin analog is cyclized through a lactam bond between R2and R7. In still further embodiments, R1is Nle, R3is selected from dPhe, dBip, and dTrp, R5is Arg, and R6is Trp. In some embodiments, the sequence of any one of Formulae (l)-(ID) is selected from the group consisting of: Ac-Nle-c[Asp-dPhe-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 16);Ac-Nle-c[Asp-dBip-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 17); andAc-Nle-c[Asp-dTrp-p(F)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 18), represents cyclization through R2and R7via a lactam bond.

[0110] In some embodiments of the sequence of any one of Formulae (l)-(ID), R4is p(CI)dPhe. In further embodiments, the non-naturally occurring melanocortin analog is cyclized through a lactam bond between R2and R7. In still further embodiments, R1is Nle, R3is selected from dGIn, dBip, and dTrp, R5is Arg, and R6is Trp. In some embodiments, the sequence of any one of Formulae (l)-(ID) is selected from the group consisting of: Ac-Nle-c[Asp-dBip-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 11 );Ac-Nle-c[Asp-dGln-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 12); andAc-Nle-c[Asp-dTrp-p(CI)dPhe-Arg-Trp-Lys]-dVal-dPro-NH2(SEQ ID NO: 13), represents cyclization through R2and R7via a lactam bond.

[0111] In some embodiments of the sequence of any one of Formulae (l)-(ID), R4is p(l)dPhe or dBip. In further embodiments, R6is Tic. In still further embodiments, X1is Nle, R2is selected from His, Pro, and Arg, R3is Arg or Pro, R5is Arg. In some embodiments, the sequence of any one of Formulae (l)-(ID) is selected from the group consisting of: Ac-Nle-c[Asp-His-Arg-p(l)dPhe-Arg-Tic-Lys]-dVal-dPro-NH2(SEQ ID NO: 5);Ac-Nle-c[Asp-His-Arg-dBip-Arg-Tic-Lys]-dVal-dPro-NH2(SEQ ID NO: 6);Ac-Nle-c[Asp-Pro-Arg-p(l)dPhe-Arg-Tic-Lys]-dVal-dPro-NH2(SEQ ID NO: 7);Ac-Nle-c[Asp-Pro-Arg-dBip-Arg-Tic-Lys]-dVal-dPro-NH2(SEQ ID NO: 8);Ac-Nle-c[Asp-Arg-Pro-p(l)dPhe-Arg-Tic-Lys]-dVal-dPro-NH2(SEQ ID NO: 9); andAc-Nle-c[Asp-Arg-Pro-dBip-Arg-Tic-Lys]-dVal-dPro-NH2(SEQ ID NO: 10), represents cyclization through R1and R7via a lactam bond.

[0112] In some embodiments of the sequence of any one of Formulae (l)-(ID), R4is dNal(2’). In further embodiments, the non-naturally occurring melanocortin analog is cyclized through a disulfide bond between dPen at R1or R2and dPen at R7In still further embodiments, X1is Nle or absent, R1is Nle or dPen, R3is selected from Ala, Leu, dLeu, dVal, and Gly, R5is Arg, and R6is Trp. In some embodiments, the sequence of any one of Formulae (l)-(ID) is selected from the group consisting of:Ac-Nle-c[dPen-Ala-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 25);Ac-Nle-c[dPen-Leu-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 26);Ac-Nle-c[dPen-dLeu-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 27);Ac-Nle-c[dPen-dVal-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 28);Ac-Nle-c[dPen-Gly-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 29); andAc-Nle-c[dPen-Gly-Gly-dNal(2')-Arg-Trp-dPen]-dVal-dPro-NH2(SEQ ID NO: 30), represents cyclization through R1or R2and R7via a disulfide bond.

[0113] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (I). In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 2-37.

[0114] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA). In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 2-3, 14, 15, 19- 24, and 31-37.

[0115] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(i)). In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 2-3, 14, 15, and 19-24.

[0116] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(ii)). In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 2, 19-24, 35 and 37.

[0117] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(iii)). In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of SEQ ID NOs: 35 or 37.

[0118] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IB). In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 11-13 and 16- 18. In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 11 -13. In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 16-18.

[0119] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IC). In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 5-10. In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 5, 7, and 9. In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 6, 8, and 10.

[0120] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (ID). In some embodiments, the non-naturally occurring melanocortin analog comprises any one of the sequences of SEQ ID NOs: 25-30.Non-Naturally Occurring Melanocortin Analog Synthesis

[0121] The non-naturally occurring melanocortin analogs of the present technology may be readily synthesized by any known conventional procedure for the formation of a peptide linkage between amino acids. Such conventional procedures include, for example, any solution phase procedure permitting a condensation between the free alpha amino group of an amino acid or residue thereof having the carboxyl group or other reactive groups protected and the free primary carboxyl group of another amino acid or residue thereofhaving the amino group or other reactive groups protected. In an exemplary procedure, the peptides of the present technology may be synthesized by solid-phase synthesis and purified according to methods known in the art. Any of a number of well-known procedures utilizing a variety of resins and reagents may be used to prepare the peptides of the present technology.

[0122] The process for synthesizing the peptides may be carried out by a procedure whereby each amino acid in the desired sequence is added one at a time in succession to another amino acid or residue thereof or by a procedure whereby peptide fragments with the desired amino acid sequence are first synthesized conventionally and then condensed to provide the desired peptide. The resulting peptide is then cyclized to yield a cyclic peptide.

[0123] Solid phase peptide synthesis methods are well known and practiced in the art. In such methods, the synthesis of peptides may be carried out by sequentially incorporating the desired amino acid residues one at a time into the growing peptide chain according to the general principles of solid phase methods. These methods are disclosed in numerous references, including 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 (1980), the disclosure of which is incorporated herein by reference in its entirety.

[0124] In chemical syntheses of peptides, reactive side chain groups of the various amino acid residues are protected with suitable protecting groups, which prevent a chemical reaction from occurring at that site until the protecting group is removed. Also common is the protection of the alpha amino group of an amino acid residue or fragment while that entity reacts at 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 for solid phase synthesis methods and solution phase synthesis methods groups are known to those having ordinary skill in the art.

[0125] Alpha amino groups may be 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 is useful for alpha amino protection.

[0126] Guanidino groups may be protected by a suitable protecting group, such as nitro, p-toluenesulfonyl (Tosyl), Z, pentamethylchromanesulfonyl (Pmc), adamantyloxycarbonyl, pentamethyldihydrobenzofuran-5-sulfonyl (Pbf) and Boc. Pmc is a useful protecting group for Arg.

[0127] Solid phase synthesis is commenced from the C-terminal end of the peptide by coupling a protected alpha amino acid to a suitable resin. Such starting material is prepared by attaching an alpha amino-protected amino acid by an ester linkage to a p- benzyloxybenzyl alcohol (Wang) resin or a 2-chlorotrityl chloride resin, by an amide bond between an Fmoc-Linker, such as p-[(R,S)-a-[1-(9H-fluor-en-9-yl)-methoxyformamido]-2,4- dimethyloxybenzyl]-phenoxyacetic acid (Rink linker) to a benzhydrylamine (BHA) resin, or by other means well known in the art. Fmoc-Linker-BHA resin supports are commercially available and generally used when feasible. The resins are carried through repetitive cycles as necessary to add amino acids sequentially. The alpha amino Fmoc protecting groups are removed under basic conditions. Piperidine, piperazine, diethylamine, or morpholine (20- 40% v / v) in N,N-dimethylformamide (DMF) may be used for this purpose.

[0128] Following removal of the alpha amino protecting group, the subsequent protected amino acids are coupled stepwise in the desired order to obtain an intermediate, protected peptide-resin. The activating reagents used for coupling of the amino acids in the solid phase synthesis of the peptides are well known in the art. After the peptide is synthesized, if desired, the orthogonally protected side chain protecting groups may be removed using methods well known in the art for further derivatization of the peptide.

[0129] Reactive groups in a peptide may be selectively modified, either during solid phase synthesis or after removal from the resin. For example, peptides may be modified to obtain N-terminus modifications, such as acetylation, while on resin, or may be removed from the resin by use of a cleaving reagent and then modified. Methods for N-terminus modification, such as acetylation, and for C-terminus modification, such as amidation, are known in the art. Similarly, methods for modifying side chains of amino acids are well knownto those skilled in the art of peptide synthesis. The choice of modifications made to reactive groups present on the peptide will be determined, in part, by the characteristics that are desired in the peptide.

[0130] The peptide may be cyclized prior to cleavage from the peptide resin. For cyclization through reactive side chain moieties, the desired side chains are deprotected, and the peptide suspended in a suitable solvent and a cyclic coupling agent added. Suitable solvents include, for example DMF, dichloromethane (DCM) or 1 -methyl-2-pyrrolidone (NMP). Suitable cyclic coupling reagents include, for example, 2-(1 H-benzotriazol-1 -yl)- 1 ,1 ,3,3-tetramethyluronium tetrafluoroborate (TBTU), 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 (TATLI), 2-(2-oxo-1 (2H)-pyridyl)-1 , 1 ,3,3- tetramethyluronium tetrafluoroborate (TPTU) or N,N'-dicyclohexylcarbodiimide / 1- hydroxybenzotriazole (DCCI / HOBt). Coupling is convention initiated by use of a suitable base, such as N,N-diispropylethylamine (DIPEA), sym-collidine, or N-methylmorpholine (NMM).

[0131] Following cleavage of peptides from the solid phase following their synthesis, the peptide may be purified by any number of methods, such as reverse phase high performance liquid chromatography (RP-HPLC), using a suitable column, such as a C18 column. Other methods of separation or purification, such as methods based on the size or charge of the peptide, may also be employed. Once purified, the peptide may be characterized by any number of methods, such as high performance liquid chromatograph (HPLC), amino acid analysis, mass spectrometry, and the like.Salt Forms of Non-Naturallv Occurring Melanocortin Analogs

[0132] The non-naturally occurring melanocortin analog peptides of the present technology may be in the form of any salt. The term “pharmaceutically acceptable salts” refers to salts prepared from non-toxic bases or acids including inorganic or organic bases and inorganic or organic acids. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts,manganous, potassium, sodium, zinc, and the like. Exemplary salts are the ammonium, calcium, lithium, magnesium, potassium, and sodium salts. Salts derived from organic nontoxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

[0133] When the non-naturally occurring melanocortin analogs of the present technology are basic, acid addition salts may be prepared from non-toxic acids, including inorganic and organic acids. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, carboxylic, citric, ethanesulfonic, formic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, malonic, mucic, nitric, pamoic, pantothenic, phosphoric, propionic, succinic, sulfuric, tartaric, p-toluenesulfonic acid, trifluoroacetic acid, and the like. Acid addition salts of the peptides of the present technology are prepared in a suitable solvent from the peptide and an excess of an acid, such as hydrochloric, hydrobromic, sulfuric, phosphoric, acetic, trifluoroacetic, citric, tartaric, maleic, succinic or methanesulfonic acid. The acetate salt form is especially useful. Where the peptides of the present technology include an acidic moiety, suitable salts may include alkali metal salts, such as sodium or potassium salts, or alkaline earth metal salts, such as calcium or magnesium salts.Conjugates

[0134] The present technology further includes conjugates comprising a non-naturally occurring melanocortin analog. In some embodiments, the non-naturally melanocortin analog is conjugated to a pharmaceutical agent. Non-limiting examples of suitable pharmaceutical agents include peptides, monoclonal antibodies, and small molecules. In some embodiments, the non-naturally occurring melanocortin analog is conjugated to semaglutide. In other embodiments, the non-naturally occurring melanocortin analog isconjugated to setmelanotide. In yet other embodiments, the non-naturally occurring melanocortin analog is conjugated to a small molecule MCR agonist.

[0135] Conjugates of the present technology further comprise a linker connecting the non-naturally occurring melanocortin analog to the pharmaceutical agent. In some embodiments, the linker is rigid. In some embodiments, the linker is flexible. In some embodiments, the linker is a cleavable linker. In some embodiments, the linker comprises a glycine-serine (Gly / Ser) linker, a proline-threonine-glycine linker, an alanine linker, a lysine linker, a threonine linker, a valine-glycine-serine-threonine linker, an elastin-like peptide linker, a hexahistidine linker, a polyethylene glycol linker, a fatty acid linker, or a hydrocarbon linker.

[0136] In some embodiments, the linker is a peptide linker. The peptide linkers of the present technology may vary from 2 to 31 amino acids of any primary sequence in length and do not impose any constraints on the conformation or interactions of the linked partners. In some embodiments, the linkers vary from about 2-30, 2-29, 2-28, 2-27, 2-26, 2-25, 2-24,2-23, 2-22, 2-21 , 2-20, 2-19, 2-18, 2-17, 2-16, 2-15, 2- 14, 2-13, 2-12, 2-11 , 2-10, 2-9, 2-8,2-7, 2-6, 2-5, 2-4, 2-3, 3-31 , 3-30, 3-29, 3-28, 3-27, 3-26, 3-25, 3-24, 3-23, 3-22, 3-21 , 3-20,3-19, 3-18, 3-17, 3-16, 3-15, 3-14, 3-13, 3-12, 3-11 , 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-31 ,4-30, 4-29, 4-28, 4-27, 4-26, 4-25, 4-24, 4-23, 4-22, 4-21 , 4- 20, 4-19, 4-18, 4-17, 4-16, 4- 15, 4-14, 4-13, 4-12, 4-11 , 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-31 , 5- 30, 5-29, 5-28, 5-27, 5-26,5-25, 5-24, 5-23, 5-22, 5-21 , 5-20, 5-19, 5-18, 5-17, 5-16, 5-15, 5- 14, 5-13, 5-12, 5-11 , 5- 10, 5-9, 5-8, 5-7, 5-6, 6-31 , 6-30, 6-29, 6-28, 6-27, 6-26, 6-25, 6-24, 6-23, 6-22, 6-21 , 6-20,6-19, 6-18, 6-17, 6-16, 6-15, 6-14, 6-13, 6-12, 6-11 , 6-10, 6-9, 6-8, 6- 7, 7-31 , 7-30, 7-29, 7- 28, 7-27, 7-26, 7-25, 7-24, 7-23, 7-22, 7-21 , 7-20, 7-19, 7-18, 7-17, 7- 16, 7-15, 7-14, 7-13,7-12, 7-11 , 7-10, 7-9, 7-8, 8-31 , 8-30, 8-29, 8-28, 8-27, 8-26, 8-25, 8-24, 8-23, 8-22, 8-21 ,8-20, 8-19, 8-18, 8-17, 8-16, 8-15, 8-14, 8-13, 8-12, 8-11 , 8-10, 8-9, 9-31 , 9-30, 9-29, 9-28,9-27, 9-26, 9-25, 9-24, 9-23, 9-22, 9-21 , 9-20, 9-19, 9-18, 9-17, 9-16, 9-15, 9-14, 9-13, 9-12,9-11 , 9-10, 10-31 , 10-30, 10-29, 10-28, 10-27, 10-26, 10-25, 10-24, 10-23, 10-22, 10-21 , I Q-20, 10-19, 10-18, 10-17, 10-16, 10-15, 10-14, 10-13, 10-12, 10-11 , 11-31 , 11-30, 11-29, 11-28, 11 -27, 11 -26, 11-25, 11-24, 11 -23, 11 -22, 11-21 , 11-20, 11 -19, 11-18, 11-17, 11-16, 11 -15, 11 -14, 11 -13, 11-12, 12-31 , 12-30, 12-29, 12-28, 12-27, 12-26, 12-25,12-24, 12-23, 12-22, 12-21 , 12-20, 12-19, 12-18, 12-17, 12-16, 12-15, 12-14, 12-13, 13-31 , 13-30, 13-29, 13-28, 13-27, 13-26, 13-25, 13-24, 13-23, 13-22, 13-21 , 13-20, 13-19, 13-18, 13-17, 13-16, 13-15, 13-14, 14-31 , 14-30, 14-29, 14-28, 14-27, 14-26, 14-25, 14-24, 14-23, 14-22, 14-21 , 14-20, 14-19, 14-18, 14-17, 14-16, 14-15, 15-31 , 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21 , 15-20, 15-19, 15-18, 15-17, 15-16, 16-31 , 16-30, 16-29, 16-28, 16-27, 16-26, 16-25, 16-24, 16-23, 16-22, 16-21 , 16-20, 16-19, 16-18, 16-17, 17-31 , 17-30, 17-29, 17-28, 17-27, 17-26, 17-25, 17-24, 17-23, 17-22, 17-21 , 17-20, 17-19, 17-18, 18-31 , 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21 , 18-20, 18-19, 19-31 , 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21 , 19-20, 20-31 , 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21 , 21 -31 , 21-30, 21-29, 21-28, 21-27, 21 -26, 21 -25, 21-24, 21-23, 21 -22, 22-31 , 22-30, 22-29, 22-28, 22-27, 22-26, 22-25, 22-24, 22-23, 23-31 , 23-30, 23-29, 23-28, 23-27, 23-26, 23-25, 23-24, 24-31 , 24-30, 24-29, 24-28, 24-27, 24-26, 24-25, 25-31 , 25-30, 25-29, 25-28, 25-27, 25-26, 26-31 , 26-30, 26-29, 26-28, 26-27, 27-31 , 27-30, 27-29, 27-28, 28-31 , 28-30, 28-29, 29-31 , 29-30, or 30-31 amino acids of any primary sequence in length. The peptide linkers may be designed as appropriate for an intended use.

[0137] The peptide linkers may comprise one or more of a Gly-rich linker (e.g., a flexible linker connecting various domains in a single protein without interfering with the function of each domain; a linker forming stable covalently linked dimers; a linker to connect two independent domains that create a ligand-binding site or recognition sequence), a Serine linker (e.g., a coiled structure linker); a coiled structure linker comprising a Gin, Arg, Glu, Ser, and / or Pro amino acids; a rigid space linker comprising one or more of a Pro, Arg, Phe, Thr, Glu, and / or Gin residues; a linker comprising a flexible Gly-rich regions that may may generate loops connecting domains; or a linker comprising a Thr, Ser, Gly, and / or Ala residue.

[0138] In some embodiments, the linker comprises an amino acid sequence about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 100% identical to the amino acid sequence of any one of Linker A, Linker B, Linker C, Linker D, Linker E, Linker F, Linker G, Linker H, Linker I, Linker J, Linker K, Linker L, Linker M, Linker N, Linker O, Linker P, Linker Q, Linker R, or Linker S in Table 0.-SO-

[0139] In some embodiments, the linker comprises an amino acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 100% identical to the amino acid sequence of any one of Linker A, Linker B, Linker C, Linker D, Linker E, Linker F, Linker G, Linker H, Linker I, Linker J, Linker K, Linker L, Linker M, Linker N, Linker 0, Linker P, Linker Q, Linker R, or Linker S in Table 0.In some embodiments, the linker comprises an amino acid sequence at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 100% identical to the amino acid sequence of any one of Linker A, Linker B, Linker C, Linker D, Linker E, Linker F, Linker G, Linker H, Linker I, Linker J, Linker K, Linker L, Linker M, Linker N, Linker O, Linker P, Linker Q, Linker R, or Linker S in Table 0.Table 0. Peptide linkersPharmaceutical Compositions

[0140] In some embodiments, the non-naturally occurring melanocortin analogs of the present technology are present in a pharmaceutical composition.

[0141] In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in a concentration of 0.1 mg / mL to 500 mg / mL, relative to a total volume of the composition. For example, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in a concentration of 0.1 mg / mL to 500 mg / mL, 0.5 mg / mL to 250 mg / mL, 1 mg / mL to 100 mg / mL, 2.5 mg / mL to 50 mg / mL, or 5 mg / mL to 25 mg / mL, relative to a total volume of the composition. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in a concentration of about 50 mg / mL, relative to a total volume of the pharmaceutical composition.

[0142] In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of any one of Formulae (l)-(IC), and the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in a concentration of 5 mg / mL to 100 mg / mL, relative to a total volume of the pharmaceutical composition. For example, the non-naturally occurring melanocortin analog comprising a sequence of any one of Formulae (l)-(IC) is present in the pharmaceutical composition in a concentration of 5 mg / mL to 100 mg / mL, 10 mg / mL to 75 mg / mL, 15 mg / mL to 50 mg / mL, 20 mg / mL to 40 mg / mL, or 25 mg / mL to 30 mg / mL, relative to a total volume of the pharmaceutical composition. In some embodiments, the non-naturally occurring melanocortin analog comprising a sequence of any one of Formulae (l)-(IC) is present in the pharmaceutical composition in a concentration of about 50 mg / mL, relative to a total volume of the pharmaceutical composition.

[0143] In some embodiments, the composition comprises the non-naturally occurring melanocortin analog of a sequence of any one of Formulae (l)-(IC) at a concentration atabout 0.001 nmol, 0.005 nmol, 0.01 nmol, 0.02 nmol, 0.05 nmol, 0.1 nmol, 0.25 nmol, 0.5 nmol, 1 nmol, 2.5 nmol, 5 nmol, 10 nmol, 20 nmol, 25 nmol, 50 nmol, 100 nmol, 250 nmol, 500 nmol, or 1000 nmol, or more, depending on the specific peptide selected, the desired response, the route of administration, the formulation and other factors known to those of skill in the art. In some embodiments, the non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(iii)).

[0144] The non-naturally occurring melanocortin analogs may be formulated with one or more pharmaceutically acceptable carriers and / or excipients. The carriers and / or excipients of the present technology facilitate delivery of the non-naturally occurring melanocortin analog to a subject. Other pharmaceutically acceptable carriers and / or excipients may be included in the pharmaceutical composition to enhance dispersion, solubility, and / or stability of the non-naturally occurring melanocortin analog, and / or to reduce adverse injection site reactions.

[0145] In some embodiments, the pharmaceutical composition comprises 0.1 to 99.9999 wt.%, 1 to 99.999 wt.%, 5 to 99.99 wt.%, 10 to 99.9 wt.%, 15 to 99 wt.%, 20 to 90 wt.%, 30 to 85 wt.%, 40 to 80 wt.%, 50 to 75 wt.%, or 60 to 70 wt.% of the pharmaceutically acceptable carrier and / or excipient relative to a total weight of the pharmaceutical composition.

[0146] In some embodiments, the pharmaceutically acceptable carrier and / or excipient of the pharmaceutical composition is selected from the group consisting of water, a buffer, an inorganic salt, a fatty acid, a vegetable oil, a synthetic fatty ester, a surfactant, and a polymer.

[0147] In some embodiments, the pharmaceutically acceptable carrier and / or excipient of the pharmaceutical composition is water. In some embodiments, the pharmaceutically acceptable carrier and / or excipient of the pharmaceutical composition is a buffer.

[0148] In some embodiments, the pharmaceutical composition contains about 0.1 to about 99.9999 wt.%, about 1 to about 99.999 wt.%, about 5 to about 99.99 wt.%, about 10 to about 99.9 wt.%, about 15 to about 99 wt.%, about 20 to about 90 wt.%, about 30 to about 85 wt.%, about 40 to about 80 wt.%, about 50 to about 75 wt.%, or about 60 to about 70wt.% of one or more pharmaceutically acceptable carriers and / or excipients relative to a total weight of the pharmaceutical composition.

[0149] In some embodiments, the pharmaceutical composition contains at least 0.1 to at least 99.9999 wt.%, at least 1 to at least 99.999 wt.%, at least 5 to at least 99.99 wt.%, at least 10 to at least 99.9 wt.%, at least 15 to at least 99 wt.%, at least 20 to at least 90 wt.%, at least 30 to at least 85 wt.%, at least 40 to at least 80 wt.%, at least 50 to at least 75 wt.%, or at least 60 to at least 70 wt.% of one or more pharmaceutically acceptable carriers and / or excipients relative to a total weight of the pharmaceutical composition.

[0150] In some embodiments, the pharmaceutical composition contains at least about 0.1 to at least about 99.9999 wt.%, at least about 1 to at least about 99.999 wt.%, at least about 5 to at least about 99.99 wt.%, at least about 10 to at least about 99.9 wt.%, at least about 15 to at least about 99 wt.%, at least about 20 to at least about 90 wt.%, at least about 30 to at least about 85 wt.%, at least about 40 to at least about 80 wt.%, at least about 50 to at least about 75 wt.%, or at least about 60 to at least about 70 wt.% of one or more pharmaceutically acceptable carriers and / or excipients relative to a total weight of the pharmaceutical composition.

[0151] Any pharmaceutically acceptable carriers and / or excipients known in the art may be included in the pharmaceutical composition. Non-limiting examples of pharmaceutically acceptable carriers and / or excipients include buffers, binders, excipients, stabilizers, lubricants, oils, adjuvants, preservatives, lipids, and antioxidants. The pharmaceutical composition may comprise any combination of the one or more pharmaceutically acceptable carriers and / or excipients previously described in relation to the first and pharmaceutical compositions. In some embodiments, the one or more pharmaceutically acceptable carriers and / or excipients comprise water.

[0152] The carriers and / or excipients of the composition may generally include one or more of the following components: (i) one or more antioxidants, (ii) one or more preservatives, (iii) one or more buffers, (iv) one or more tonicity adjustors, (v) one or more surfactants, (vi) flavor, (vii) propellants, and / or (viii) a vehicle or solvent. In some embodiments, all components are compatible with the non-naturally occurring melanocortinanalog (i.e. , do not react or cause the non-naturally occurring melanocortin analog to react) and are homogeneously dispersed or dissolved uniformly in the composition.

[0153] In some embodiments, the one or more pharmaceutically acceptable carriers and / or excipients are isotonic. In some embodiments, the carrier and / or excipient is isotonic to nasal fluids.

[0154] In some embodiments, the pharmaceutical composition further comprises a pharmaceutical salt. Any pharmaceutical salt known in the art may be included in the pharmaceutical composition. For examples, to achieve a desirable tonicity, the pharmaceutical composition may include a salt selected from the group consisting of sodium chloride, sodium succinate, sodium sulfate, potassium chloride, magnesium chloride, magnesium sulfate, and calcium chloride.

[0155] In some embodiments, the salt is present in the pharmaceutical composition in a concentration of about 0.1 mg / mL to about 50 mg / mL, about 1 mg / mL to about 25 mg / mL, or about 5 mg / mL to about 10 mg / mL, relative to a total volume of the composition.

[0156] In some embodiments, the salt is present in the pharmaceutical composition in a concentration of at least 0.1 mg / mL to at least 50 mg / mL, at least 1 mg / mL to at least 25 mg / mL, or at least 5 mg / mL to at least 10 mg / mL, relative to a total volume of the composition.

[0157] In some embodiments, the salt is present in the pharmaceutical composition in a concentration of at least about 0.1 mg / mL to at least about 50 mg / mL, at least about 1 mg / mL to at least about 25 mg / mL, or at least about 5 mg / mL to at least about 10 mg / mL, relative to a total volume of the composition.

[0158] Pharmaceutically acceptable carriers and / or excipients that may be included in the pharmaceutical composition generally include a pH buffered aqueous solution comprising one or more of the following components: (a) sodium acetate, (b) Ths, and (c) water. In some embodiments, all components are compatible with the non-naturally occurring melanocortin analog (i.e., do not react or cause the non-naturally occurring melanocortin analog to react) and are homogeneously dispersed or dissolved uniformly in the composition.

[0159] In the pH buffered solution of the pharmaceutical composition, the water may act as a diluent and include, without limitation, water for injection (WFI), sterile water, bacteriostatic water for injection (BWFI), distilled water, bidistilled water, deionized water, deionized distilled water, and reverse osmosis water. In some embodiments, the water present in the pH buffered aqueous solution is water for injection.

[0160] In some embodiments, the pharmaceutical composition includes water in an amount of about 1 wt% to about 90 wt%, about 10 wt% to about 75 wt%, or about 25 wt% to about 50 wt%, relative to a total weight of the composition.

[0161] In some embodiments, the pharmaceutical composition includes water in an amount of at least 1 wt% to at least 90 wt%, at least 10 wt% to at least 75 wt%, or at least 25 wt% to at least 50 wt%, relative to a total weight of the composition.

[0162] In some embodiments, the pharmaceutical composition includes water in an amount of at least about 1 wt% to at least about 90 wt%, at least about 10 wt% to at least about 75 wt%, or at least about 25 wt% to at least about 50 wt%, relative to a total weight of the composition.

[0163] In some embodiments, sodium acetate is present in the pharmaceutical composition in a concentration of 0.5 mg / mL to 50 mg / mL, relative to a total volume of the composition. For example, sodium acetate may be present in the pharmaceutical composition in a concentration of 0.5 mg / mL to 50 mg / mL, 1 mg / mL to 40 mg / mL, 2 mg / mL to 30 mg / mL, 4 mg / mL to 20 mg / mL, 5 mg / mL to 15 mg / mL, 6 mg / mL to 12 mg / mL, or 8 mg / mL to 10 mg / mL, relative to a total volume of the composition.

[0164] In some embodiments, sodium acetate is present in the pharmaceutical composition in a concentration of about 6 mg / mL to about 8 mg / mL, relative to a total volume of the composition. For example, sodium acetate may be present in the pharmaceutical composition in a concentration of 6 mg / mL, 6.5 mg / mL, 7 mg / mL, 7.1 mg / mL, 7.5 mg / mL, or 8 mg / mL, relative to a total volume of the composition.

[0165] In some embodiments, sodium acetate is present in the pharmaceutical composition in a molar concentration of 5 mM to 700 mM, relative to a total volume of the composition. For example, sodium acetate may be present in the pharmaceuticalcomposition in a molar concentration of 5 mM to 700 mM, 10 mM to 600 mM, 20 mM to 500 mM, 30 mM to 400 mM, 40 mM to 300 mM, 50 mM to 200 mM, 60 mM to 100 mM, or 70 mM to 80 mM, relative to a total volume of the composition.

[0166] In some embodiments, sodium acetate is present in the pharmaceutical composition in a molar concentration of about 80 mM to about 100 mM, relative to a total volume of the composition. For example, sodium acetate may be present in the pharmaceutical composition in a molar concentration of 80 mM, 85 mM, 87 mM, 90 mM, 95 mM, or 100 mM, relative to a total volume of the composition.

[0167] The term “Tris” refers to tris(hydroxymethyl)aminomethane, which is also known as Tris buffer, Tris base, TRIS, tromethamine, tromethamine buffer, Trizma®, Trisamine, Trometamol, Tromethane, Trisaminol, or THAM. In some embodiments, Tris is present in the pharmaceutical composition in a concentration of 0.5 mg / mL to 50 mg / mL, relative to a total volume of the composition. For example, Tris may be present in the pharmaceutical composition in a concentration of 0.5 mg / mL to 50 mg / mL, 1 mg / mL to 40 mg / mL, 2 mg / mL to 30 mg / mL, 4 mg / mL to 20 mg / mL, 5 mg / mL to 15 mg / mL, 6 mg / mL to 12 mg / mL, or 8 mg / mL to 10 mg / mL, relative to a total volume of the composition.

[0168] In some embodiments, Tris is present in the pharmaceutical composition in a concentration of about 6 mg / mL to about 8 mg / mL, relative to a total volume of the composition. For example, Tris may be present in the pharmaceutical composition in a concentration of 6 mg / mL, 6.5 mg / mL, 7 mg / mL, 7.3 mg / mL, 7.6 mg / mL, or 8 mg / mL, relative to a total volume of the composition.

[0169] In some embodiments, Tris is present in the pharmaceutical composition in a molar concentration of 2 mM to 500 mM, relative to a total volume of the composition. For example, Tris may be present in the pharmaceutical composition in a molar concentration of 2 mM to 500 mM, 5 mM to 400 mM, 10 mM to 300 mM, 20 mM to 200 mM, 30 mM to 150 mM, 40 mM to 100 mM, 50 mM to 80 mM, or 60 mM to 70 mM, relative to a total volume of the composition.

[0170] In some embodiments, Tris is present in the pharmaceutical composition in a molar concentration of about 50 mM to about 70 mM, relative to a total volume of thecomposition. For example, Tris may be present in the pharmaceutical composition in a molar concentration of 50 mM, 55 mM, 60 mM, 65 mM, or 70 mM, relative to a total volume of the composition.

[0171] In some embodiments, the pH buffered aqueous solution provides the pharmaceutical composition with a pH equivalent or close to the physiological pH levels. This may reduce adverse injection site reactions and also provide the non-naturally occurring melanocortin analog with enhanced stability and resistance to aggregation and degradation.

[0172] In some embodiments, a weight ratio of sodium acetate to Tris is about 1 :4 to about 4:1 , about 2:7 to about 7:2, about 1 :3 to about 3:1 , about 2:5 to about 5:2, about 1 :2 to about 2:1 , about 2:3 to about 3:2, or about 1 :1. In some embodiments, the weight ratio of sodium acetate to Tris is about 1 :1.

[0173] In some embodiments, a weight ratio of sodium acetate to Tris is at least 1 :4 to at least 4:1 , at least 2:7 to at least 7:2, at least 1 :3 to at least 3:1 , at least 2:5 to at least 5:2, at least 1 :2 to at least 2:1 , at least 2:3 to at least 3:2, or at least 1 :1. In some embodiments, the weight ratio of sodium acetate to Tris is at least 1 :1.

[0174] In some embodiments, a weight ratio of sodium acetate to Tris is at least about 1 :4 to at least about 4:1 , at least about 2:7 to at least about 7:2, at least about 1 :3 to at least about 3:1 , at least about 2:5 to at least about 5:2, at least about 1 :2 to at least about 2:1 , at least about 2:3 to at least about 3:2, or at least about 1 :1. In some embodiments, the weight ratio of sodium acetate to Tris is at least about 1 :1.

[0175] In some embodiments, a weight ratio of the non-naturally occurring melanocortin analog to sodium acetate is about 1 :1 to about 20:1 , about 3:2 to about 15:1 , about 2:1 to about 12:1 , about 3:1 to about 10:1 , about 4:1 to about 9:1 , about 5:1 to about 8:1 , or about 6:1 to about 7:1. In some embodiments, the weight ratio of the non-naturally occurring melanocortin analog to sodium acetate is about 7:1 .

[0176] In some embodiments, a weight ratio of the non-naturally occurring melanocortin analog to sodium acetate is at least 1 :1 to at least 20:1 , at least 3:2 to at least 15:1 , at least 2:1 to at least 12:1 , at least 3: 1 to at least 10: 1 , at least 4: 1 to at least 9:1 , atleast 5:1 to at least 8:1 , or at least 6:1 to at least 7:1 . In some embodiments, the weight ratio of the non-naturally occurring melanocortin analog to sodium acetate is at least 7:1.

[0177] In some embodiments, a weight ratio of the non-naturally occurring melanocortin analog to sodium acetate is at least about 1 :1 to at least about 20:1 , at least about 3:2 to at least about 15:1 , at least about 2:1 to at least about 12:1 , at least about 3:1 to at least about 10:1 , at least about 4:1 to at least about 9:1 , at least about 5:1 to at least about 8:1 , or at least about 6:1 to at least about 7:1 . In some embodiments, the weight ratio of the non-naturally occurring melanocortin analog to sodium acetate is at least about 7:1.

[0178] In some embodiments, a weight ratio of the non-naturally occurring melanocortin analog to T ris is about 1 : 1 to about 20: 1 , about 3:2 to about 15:1 , about 2: 1 to about 12:1 , about 3: 1 to about 10:1 , about 4: 1 to about 9: 1 , about 5: 1 to about 8:1 , or about 6:1 to about 7:1. In some embodiments, the weight ratio of the non-naturally occurring melanocortin analog to Tris is about 7:1 .

[0179] In some embodiments, a weight ratio of the non-naturally occurring melanocortin analog to Tris is at least 1 :1 to at least 20:1 , at least 3:2 to at least 15:1 , at least 2:1 to at least 12:1 , at least 3:1 to at least 10:1 , at least 4:1 to at least 9:1 , at least 5:1 to at least 8:1 , or at least 6:1 to at least 7:1. In some embodiments, the weight ratio of the non-naturally occurring melanocortin analog to Tris is at least 7:1.

[0180] In some embodiments, a weight ratio of the non-naturally occurring melanocortin analog to Tris is at least about 1 :1 to at least about 20:1 , at least about 3:2 to at least about 15:1 , at least about 2:1 to at least about 12:1 , at least about 3:1 to at least about 10:1 , at least about 4:1 to at least about 9:1 , at least about 5:1 to at least about 8:1 , or at least about 6:1 to at least about 7:1. In some embodiments, the weight ratio of the non- naturally occurring melanocortin analog to Tris is at least about 7:1.

[0181] In addition to sodium acetate and Tris, the pharmaceutical composition may include other buffering agents. Non-limiting examples of additional buffering agents include saline, phosphate, phosphoric acid, citrate, succinate, gluconate, histidine, acetic acid, ascorbate, tartartic acid, maleic acid, glycine, lactate, lactic acid, ascorbic acid, imidazole, bicarbonate, carbonic acid, succinic acid, sodium benzoate, benzoic acid, gluconate,edetate, malate, imidazole, and mixtures thereof. In some embodiments, the pharmaceutical composition comprises acetic acid as an additional buffering agent.

[0182] The pharmaceutical composition may further comprise one or more chelating agents. Suitable chelating agents include, but are not limited to edetate disodium dihydrate, calcium disodium edetate, sodium edetate, calcium versetamide sodium, calteridol, and diethylenetriaminepentaacetic acid. In some embodiments, the pharmaceutical composition further comprises edetate disodium dihydrate.

[0183] The pharmaceutical composition may further comprise a preservative agent. Exemplary preservative agents include, but are not limited to, ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, phenol, m-cresol, benzyl alcohol, alpha-tocopherol, citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, benzalkonium chloride, phenoxyethanol, and methyl paraben. In some embodiments, when the pharmaceutical composition comprises a preservative agent, the preservative agent is phenol, benzyl alcohol, or a combination thereof.

[0184] If present in the pharmaceutical composition, the concentration of the preservative agent may range from 0.001 mg / mL to 50 mg / mL, 0.01 mg / mL to 25 mg / mL, 0.1 mg / mL to 10 mg / mL, or 1 mg / mL to 5 mg / mL, relative to a total volume of the composition.

[0185] The pharmaceutical composition may further comprise an emulsifier. Nonlimiting examples of emulsifiers that may be included in the pharmaceutical composition include sodium carboxymethylcellulose, cetyl alcohol, glycerol monostearate, methylcellulose, and stearic acid. In some embodiments, when the pharmaceutical composition comprises an emulsifier, the emulsifier is sodium carboxymethylcellulose.

[0186] The pharmaceutical composition may further comprise a lipid. Lipids may enhance solubility and / or improve permeability of the non-naturally occurring melanocortin analog. In some embodiments, the lipid is a phospholipid. Non-limiting examples of phospholipids that may be included in the pharmaceutical composition include eggphosphatidylcholine, hydrogenated soybean phoshphaditylcholine, glycerophosphocholine, lecithin, and N-(carbonyl-methoxypolyethylene glycol 2000)-1 ,2-distearoyl-glycero-3- phosphoethanolamine sodium salt. In some embodiments, when the pharmaceutical composition comprises a lipid, the lipid is N-(carbonyl-methoxypolyethylene glycol 2000)- 1 ,2-distearoyl-glycero-3-phosphoethanolamine sodium salt

[0187] The pharmaceutical composition may further comprise a bulking agent. Inclusion of a bulking agent may increase the stability of the pharmaceutical composition. Non-limiting examples of bulking agents that may be included in the pharmaceutical composition include sucrose, lactose, trehalose, mannitol, sorbitol, glucose, raffinose, glycine, histidine, and polyvinyl pyrrolidone. In some embodiments, when the pharmaceutical composition comprises a bulking agent, the bulking agent is mannitol.

[0188] In some embodiments, the pharmaceutical composition is in the form of an aqueous solution or a suspension. In some embodiments, the pharmaceutical composition is in the form of an emulsion. In some embodiments, the pharmaceutical composition is in the form of an aqueous solution. In some embodiments, the pharmaceutical composition is in the form of an aqueous solution which is clear, colorless, and / or free of visible foreign matter.

[0189] In some embodiments, the pharmaceutical composition has a pH ranging from about 6.5 to about 8.5. In some embodiments, the pharmaceutical composition has a pH of about 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1 , 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1 , 8.2, 8.3, 8.4, or 8.5.

[0190] In some embodiments, the pharmaceutical composition has a pH ranging from at least 6.5 to at least 8.5. In some embodiments, the pharmaceutical composition has a pH of at least 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1 , 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1 , 8.2, 8.3, 8.4, or 8.5.

[0191] In some embodiments, the pharmaceutical composition has a pH ranging from at least about 6.5 to at least about 8.5. In some embodiments, the pharmaceutical composition has a pH of at least about 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1 , 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1 , 8.2, 8.3, 8.4, or 8.5.

[0192] In some embodiments, the pharmaceutical composition is basic and has a pH of about 7.1 , 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1 , 8.2, 8.3, 8.4, or 8.5. In some embodiments, the pharmaceutical composition has a pH ranging from about 7.3 to about 7.4. In some embodiments, the pharmaceutical composition has a pH of 7.3 or 7.4.

[0193] In some embodiments, the pharmaceutical composition is basic and has a pH of at least 7.1 , 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1 , 8.2, 8.3, 8.4, or 8.5. In some embodiments, the pharmaceutical composition has a pH ranging from at least 7.3 to at least 7.4. In some embodiments, the pharmaceutical composition has a pH of 7.3 or 7.4.

[0194] In some embodiments, the pharmaceutical composition is basic and has a pH of at least about 7.1 , 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1 , 8.2, 8.3, 8.4, or 8.5. In some embodiments, the pharmaceutical composition has a pH ranging from at least about 7.3 to at least about 7.4. In some embodiments, the pharmaceutical composition has a pH of 7.3 or 7.4.

[0195] In some embodiments, the pharmaceutical composition has an osmolality ranging from 250 mOsm / kg to 350 mOsm / kg. For example, the pharmaceutical composition may have an osmolality ranging from 250 mOsm / kg to 360 mOsm / kg, 260 mOsm / kg to 340 mOsm / kg, 270 mOsm / kg to 330 mOsm / kg, 280 mOsm / kg to 320 mOsm / kg, 290 mOsm / kg to 310 mOsm / kg, or about 300 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolarity of about 250 mOsm / kg, about 260 mOsm / kg, about 270 mOsm / kg, about 280 mOsm / kg, about 290 mOsm / kg, about 300 mOsm / kg, about 310 mOsm / kg, about 320 mOsm / kg, about 330 mOsm / kg, about 340 mOsm / kg, about 350 mOsm / kg, or about 360 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolality ranging from about 275 mOsm / kg to about 330 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolality of about 279 mOsm / kg, about 314 mOsm / kg, or about 329 mOsm / kg.

[0196] In some embodiments, the pharmaceutical composition has an osmolality ranging from 250 mOsm / kg to 350 mOsm / kg. For example, the pharmaceutical composition may have an osmolality ranging from 250 mOsm / kg to 360 mOsm / kg, 260 mOsm / kg to 340 mOsm / kg, 270 mOsm / kg to 330 mOsm / kg, 280 mOsm / kg to 320 mOsm / kg, 290 mOsm / kg to 310 mOsm / kg, or at least 300 mOsm / kg. In some embodiments, the pharmaceuticalcomposition has an osmolarity of at least 250 mOsm / kg, at least 260 mOsm / kg, at least 270 mOsm / kg, at least 280 mOsm / kg, at least 290 mOsm / kg, at least 300 mOsm / kg, at least 310 mOsm / kg, at least 320 mOsm / kg, at least 330 mOsm / kg, at least 340 mOsm / kg, at least 350 mOsm / kg, or at least 360 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolality ranging from at least 275 mOsm / kg to at least 330 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolality of at least 279 mOsm / kg, at least 314 mOsm / kg, or at least 329 mOsm / kg.

[0197] In some embodiments, the pharmaceutical composition has an osmolality ranging from 250 mOsm / kg to 350 mOsm / kg. For example, the pharmaceutical composition may have an osmolality ranging from 250 mOsm / kg to 360 mOsm / kg, 260 mOsm / kg to 340 mOsm / kg, 270 mOsm / kg to 330 mOsm / kg, 280 mOsm / kg to 320 mOsm / kg, 290 mOsm / kg to 310 mOsm / kg, or at least about 300 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolarity of at least about 250 mOsm / kg, at least about 260 mOsm / kg, at least about 270 mOsm / kg, at least about 280 mOsm / kg, at least about 290 mOsm / kg, at least about 300 mOsm / kg, at least about 310 mOsm / kg, at least about 320 mOsm / kg, at least about 330 mOsm / kg, at least about 340 mOsm / kg, at least about 350 mOsm / kg, or at least about 360 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolality ranging from at least about 275 mOsm / kg to at least about 330 mOsm / kg. In some embodiments, the pharmaceutical composition has an osmolality of at least about 279 mOsm / kg, at least about 314 mOsm / kg, or at least about 329 mOsm / kg.

[0198] In some embodiments, the pharmaceutical composition has a viscosity ranging from about 0.5 cP to about 5 cP. For example, the pharmaceutical composition may have a viscosity ranging from about 0.5 cP to about 5 cP, about 0.75 cP to about 4.5 cP, about 1 .0 cP to about 4 cP, about 1 .2 cP to about 3.5 cP, about 1 .3 cP to about 3 cP, about 1 .4 cP to about 2.5 cP, about 1.5 cP to about 2 cP, or about 1.6 cP to about 1.8 cP. In some embodiments, the pharmaceutical composition has a viscosity of about 0.5 cP, 0.6 cP, 0.7 cP, 0.8 cP, 0.9 cP, 1 .0 cP, 1 .1 cP, 1 .2 cP, 1 .3 cP, 1 .4 cP, 1 .5 cP, 1 .6 cP, 1 .7 cP, 1 .8 cP, 1 .9 cP, or 2.0 cP. In some embodiments, the pharmaceutical composition has a viscosity of about 1 .4 cP or about 1 .6 cP.

[0199] In some embodiments, the pharmaceutical composition has a viscosity ranging from at least 0.5 cP to at least 5 cP. For example, the pharmaceutical composition may have a viscosity ranging from at least 0.5 cP to at least 5 cP, at least 0.75 cP to at least 4.5 cP, at least 1 .0 cP to at least 4 cP, at least 1 .2 cP to at least 3.5 cP, at least 1 .3 cP to at least 3 cP, at least 1 .4 cP to at least 2.5 cP, at least 1 .5 cP to at least 2 cP, or at least 1 .6 cP to at least 1.8 cP. In some embodiments, the pharmaceutical composition has a viscosity of at least 0.5 cP, 0.6 cP, 0.7 cP, 0.8 cP, 0.9 cP, 1.0 cP, 1.1 cP, 1.2 cP, 1.3 cP, 1.4 cP, 1.5 cP, 1.6 cP, 1.7 cP, 1.8 cP, 1.9 cP, or 2.0 cP. In some embodiments, the pharmaceutical composition has a viscosity of at least 1 .4 cP or at least 1 .6 cP.

[0200] In some embodiments, the pharmaceutical composition has a viscosity ranging from at least about 0.5 cP to at least about 5 cP. For example, the pharmaceutical composition may have a viscosity ranging from at least about 0.5 cP to at least about 5 cP, at least about 0.75 cP to at least about 4.5 cP, at least about 1 .0 cP to at least about 4 cP, at least about 1.2 cP to at least about 3.5 cP, at least about 1 .3 cP to at least about 3 cP, at least about 1 .4 cP to at least about 2.5 cP, at least about 1 .5 cP to at least about 2 cP, or at least about 1.6 cP to at least about 1.8 cP. In some embodiments, the pharmaceutical composition has a viscosity of at least about 0.5 cP, 0.6 cP, 0.7 cP, 0.8 cP, 0.9 cP, 1.0 cP, 1.1 cP, 1.2 cP, 1.3 cP, 1.4 cP, 1.5 cP, 1.6 cP, 1.7 cP, 1.8 cP, 1.9 cP, or 2.0 cP. In some embodiments, the pharmaceutical composition has a viscosity of at least about 1 .4 cP or at least about 1 .6 cP.

[0201] In some embodiments, the pharmaceutical composition disclosed is formulated for parenteral administration, such as, for example, in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. The term “parenteral,” as used herein, includes subcutaneous, intravenous, intraperitoneal, intramuscular, and intralesional, or infusion techniques.

[0202] When the pharmaceutical composition is formulated for parenteral administration (e.g., subcutaneous administration), the active ingredient(s) (e.g., the non- naturally occurring melanocortin analog) may be dissolved or suspended in the aforementioned carrier and / or excipient. Additional aqueous or non-aqueous carriers that may facilitate dissolution of the active ingredient include, but are not limited to, ethanol,benzyl alcohol, DMSO, polyethylene glycol, propylene glycol, corn oil, cottonseed oil, peanut oil, sesame oil, and / or various buffers.

[0203] In some embodiments, the pharmaceutical composition is formulated for parenteral administration (e.g., subcutaneous administration) and comprises a non-naturally occurring melanocortin analog in a concentration of about 0.001 nmol, 0.005 nmol, 0.01 nmol, 0.02 nmol, 0.05 nmol, 0.1 nmol, 0.25 nmol, 0.5 nmol, 1 nmol, 2.5 nmol, 5 nmol, 10 nmol, 20 nmol, 25 nmol, 50 nmol, 100 nmol, 250 nmol, 500 nmol, or 1000 nmol, or even more, depending on the specific peptide selected, the desired therapeutic response, the route of administration, the formulation and other factors known to those of skill in the art.

[0204] In some embodiments, the pharmaceutical composition is formulated for parenteral administration (e.g., subcutaneous administration) and comprises sodium acetate in a concentration of about 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 76 mM, 77 mM, 78 mM, 79 mM, 80 mM, 81 mM, 82 mM, 83 mM, 84 mM, 85 mM, 86 mM, 87 mM, 88 mM, 89 mM, 90 mM, 91 mM, 92 mM, 93 mM, 94 mM, 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 105 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, or 200 mM.

[0205] In some embodiments, the pharmaceutical composition is formulated for parenteral administration (e.g., subcutaneous administration) and comprises Tris in a concentration of about 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 3 5mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, or 120 mM.

[0206] In some embodiments, the pharmaceutical composition comprises one or more antioxidants. For example, the pharmaceutical composition may comprise ascorbic acid, cysteine, sodium metabisulfite, propyl gallate, butylated hydroxytoluene, and / or butylated hydroxyanisole.

[0207] In some embodiments, the pharmaceutical composition comprises a surfactant, such as a sorbitan ester.

[0208] In some embodiments, the pharmaceutical composition comprises a flavoring or scent, such as an aromatic oil.

[0209] In some embodiments, the non-naturally occurring melanocortin analog is solubilized or suspended in a solvent or vehicle. The solvent or vehicle may be purified water, ethyl alcohol, and / or propylene glycol. In some embodiments, the pharmaceutical composition comprises between 0.03 wt% and 1 wt% melanocortin analog solubilized or suspended in a solvent or vehicle. For example, the pharmaceutical composition may comprise the non-naturally occurring melanocortin analog in an amount of about 0.03 wt%, about 0.05 wt%, about 0.1 wt%, about 0.15 wt%, about 0.2 wt%, about 0.25 wt%, about 0.3 wt%, about 0.35 wt%, about 0.4 wt%, about 0.45 wt%, about 0.5 wt%, about 0.55 wt%, about 0.6 wt%, about 0.65 wt%, about 0.7 wt%, about 0.75 wt%, about 0.8 wt%, about 0.85 wt%, about 0.9 wt%, about 0.95 wt%, or about 1 wt%.

[0210] In some embodiments, the pharmaceutical composition may comprise the non- naturally occurring melanocortin analog in an amount of at least 0.03 wt%, at least 0.05 wt%, at least 0.1 wt%, at least 0.15 wt%, at least 0.2 wt%, at least 0.25 wt%, at least 0.3 wt%, at least 0.35 wt%, at least 0.4 wt%, at least 0.45 wt%, at least 0.5 wt%, at least 0.55 wt%, at least 0.6 wt%, at least 0.65 wt%, at least 0.7 wt%, at least 0.75 wt%, at least 0.8 wt%, at least 0.85 wt%, at least 0.9 wt%, at least 0.95 wt%, or at least 1 wt%.

[0211] In some embodiments, the pharmaceutical composition may comprise the non- naturally occurring melanocortin analog in an amount of at least about 0.03 wt%, at least about 0.05 wt%, at least about 0.1 wt%, at least about 0.15 wt%, at least about 0.2 wt%, at least about 0.25 wt%, at least about 0.3 wt%, at least about 0.35 wt%, at least about 0.4 wt%, at least about 0.45 wt%, at least about 0.5 wt%, at least about 0.55 wt%, at least about 0.6 wt%, at least about 0.65 wt%, at least about 0.7 wt%, at least about 0.75 wt%, at least about 0.8 wt%, at least about 0.85 wt%, at least about 0.9 wt%, at least about 0.95 wt%, or at least about 1 wt%.

[0212] The non-naturally occurring melanocortin analogs of the present technology may be formulated for administration using any means known in the art, including orally, rectally, vaginally, ocularly, intranasally, topically, parenterally, or by injection. If administered by injection, the peptide injection may be intravenous (IV), subcutaneous (SC),intramuscular (IM), intraperitoneal (IP), intracerebroventricular (ICV), or other means known in the art. The non-naturally occurring melanocortin analog of the combination therapy may be formulated by any means known in the art, including but not limited to formulation as tablets, capsules, caplets, suspensions, powders, lyophilized preparations, suppositories, pessaries, ocular drops, skin patches, orally soluble formulations, enteric formulations, solutions sprays, aerosols and the like, and may be mixed and formulated with buffers, binders, excipients, stabilizers, lubricants, oils, adjuvants, anti-oxidants and other agents known in the art. In general, any route of administration by which the peptides are introduced across an epidermal layer of cells may be employed. Administration includes topical delivery. Administration includes delivery across the blood brain barrier. Administration includes delivery through mucous membranes, buccal administration, ophthalmic administration, oral administration, dermal administration, inhalation administration, nasal administration, urethral administration, vaginal administration, rectal administration, and the like.

[0213] In some embodiments, the pharmaceutical composition formulated for intranasal administration comprises a non-naturally occurring melanocortin analog at a concentration at about 0.001 nmol, 0.005 nmol, 0.01 nmol, 0.02 nmol, 0.05 nmol, 0.1 nmol, 0.25 nmol, 0.5 nmol, 1 nmol, 2.5 nmol, 5 nmol, 10 nmol, 20 nmol, 25 nmol, 50 nmol, 100 nmol, 250 nmol, 500 nmol, or 1000 nmol, or more, depending on the specific peptide selected, the desired therapeutic response, the route of administration, the formulation and other factors known to those of skill in the art.

[0214] In some embodiments, the pharmaceutical composition is formulated for oral administration. For example, the pharmaceutical composition may be in the form of a tablet, capsule, lozenge, pill, sachet, or any other orally deliverable form know in the art.

[0215] The composition may be formulated to be delivered by nose drop, spray device, or topical solution. In some embodiments, the pharmaceutical composition may be formulated as an aerosol, atomizer, inhalation, insufflation, metered-dose inhaler, or nebulizer. In some embodiments, the pharmaceutical composition includes a propellant, such as hydrofluoroalkane.

[0216] In some embodiments, the pharmaceutical composition may be configured to be administered using a spray device or nasal inhaler. The spray device or nasal inhalermay be configured to deliver 1 ug to 100ug per spray. In some embodiments, the spray device or nasal inhaler may be configured to deliver 1 ug to 100ug, 5ug to 90ug, 10ug to 80ug, 15ug to 70ug, 20ug to 60ug, 25ug, to 50ug, or 30ug to 40ug per spray.Dosing

[0217] In some embodiments, the non-naturally occurring melanocortin analog or a pharmaceutical composition thereof (e.g., the pharmaceutical composition) is administered hourly (such as every hour, every 2 hours, every 4 hours, every 8 hours, etc.), once a day, or twice a day. In some embodiments, the non-naturally occurring melanocortin analog is administered every morning, every evening, or every afternoon. In some embodiments, the non-naturally occurring melanocortin analog is administered before a meal, after a meal, or with a meal.

[0218] In some embodiments, the non-naturally occurring melanocortin analog or pharmaceutical composition thereof may be administered as a dosing regimen comprising once, twice, or three times daily administration on a (i) weekly; (ii) every other week; (iii) one week of therapy followed by two, three or four weeks off; (iv) two weeks of therapy followed by one, two, three or four weeks off; (v) three weeks of therapy followed by one, two, three, four or five week off; (vi) four weeks of therapy followed by one, two, three, four or five week off; (vii) five weeks of therapy followed by one, two, three, four or five week off; or (viii) monthly schedule. The (i)-(viii) schedules may be repeated 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , or 12 times or more. In some embodiments, the non-naturally occurring melanocortin analog or pharmaceutical composition thereof is administered at various dosages during the dosing regimen (e.g., a first dose with an effective amount of 10 mg / kg and a second dose with an effective amount of 5 mg / kg).

[0219] In some embodiments, the non-naturally occurring melanocortin analog or pharmaceutical composition thereof is administered once every other day, once every 2 or 3 days, once every third day, once per week, once every other week, once every third week, once every month, once every six weeks, once every other month, once every three months, once every six months, or once per year.

[0220] Administration of the non-naturally occurring melanocortin analog or pharmaceutical composition thereof at any of the dosing frequencies of the present technology may be repeated for a total of about 2 dosages, about 3 dosages, about 4 dosages, about 5 dosages, about 10 dosages, about 15 dosages, about 20 dosages, about 30 dosages, about 40 dosages, about 50 dosages or more.

[0221] Administration of the non-naturally occurring melanocortin analog or pharmaceutical composition thereof at any of the dosing frequencies of the present technology may be repeated for a total of at least 2 dosages, at least 3 dosages, at least 4 dosages, at least 5 dosages, at least 10 dosages, at least 15 dosages, at least 20 dosages, at least 30 dosages, at least 40 dosages, at least 50 dosages or more.

[0222] Administration of the non-naturally occurring melanocortin analog or pharmaceutical composition thereof at any of the dosing frequencies of the present technology may be repeated for a total of at least about 2 dosages, at least about 3 dosages, at least about 4 dosages, at least about 5 dosages, at least about 10 dosages, at least about 15 dosages, at least about 20 dosages, at least about 30 dosages, at least about 40 dosages, at least about 50 dosages or more.

[0223] In some embodiments, the frequency of dosages of the non-naturally occurring melanocortin analog or pharmaceutical composition thereof is the same during a treatment regimen. In other embodiments, the frequency of dosages of the non-naturally occurring melanocortin analog or pharmaceutical composition thereof is different during a treatment regimen. The non-naturally occurring melanocortin analog or the pharmaceutical composition thereof may be administered even less frequently. Alternatively, the dosage regimen may be decreased or increased from an initial dosing regimen for days, weeks, months, or years. In some embodiments, the dosing regimen is repeated at other intervals.

[0224] In some embodiments, the non-naturally occurring melanocortin analog is administered at least once daily in an amount ranging from 0.001 mg / kg to 25 mg / kg, 0.01 mg / kg to 20 mg / kg, 0.05 mg / kg to 15 mg / kg, 0.075 mg / kg to 10 mg / kg, 0.1 mg / kg to 8 mg / kg, 0.2 mg / kg to 6 mg / kg, 0.3 mg / kg to 4 mg / kg, 0.4 mg / kg to 2 mg / kg, or 0.5 mg / kg to 1 mg / kg per body weight of the subject. In some embodiments, the non-naturally occurring melanocortin analog is administered at least once daily in an amount ranging from about 0.5mg / kg to about 10 mg / kg, about 1 mg / kg to about 7.5 mg / kg, or about 2.5 mg / kg to about 5 mg / kg per body weight of the subject.

[0225] The methods disclosed herein may be performed on the subject for about 1 day,1 week, 1 month, 3 months, 6 months, 1 year, or 5 years. In some embodiments, the combination therapy (i.e., pharmaceutical combination) is administered to the subject for about 1 day, about 2 days, about 5 days, about 6 days, about 1 week, about 1 month, about2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, about 1 year, about 2 years, about 3 years, about 4 years, or about 5 years.

[0226] The methods disclosed herein may be performed on the subject for at least 1 day, 1 week, 1 month, 3 months, 6 months, 1 year, or 5 years. In some embodiments, the combination therapy (i.e., pharmaceutical combination) is administered to the subject for at least 1 day, at least 2 days, at least 5 days, at least 6 days, at least 1 week, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, at least 3 years, at least 4 years, or at least 5 years.

[0227] The methods disclosed herein may be performed on the subject for at least about 1 day, 1 week, 1 month, 3 months, 6 months, 1 year, or 5 years. In some embodiments, the combination therapy (i.e., pharmaceutical combination) is administered to the subject for at least about 1 day, at least about 2 days, at least about 5 days, at least about 6 days, at least about 1 week, at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 9 months, at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years.

[0228] In some embodiments, a method disclosed herein is performed on the subject for 1 day, 7 days, 14 days, 21 days, 28 days, 35 days, 40 days, 45 days, 50 days, 60 days, 75 days, 90 days, 100 days, 110 days, or 120 days.

[0229] In some embodiments, at least one active ingredient of the combination therapy is administered to a subject in a continuous doing schedule. As used herein, a “continuous dosing schedule” is an administration or dosing regimen without dose interruptions, e.g.,without days off treatment. Repetition of 21 -day or 28-day treatment cycles without dose interruptions is an exemplary continuous dosing schedule.

[0230] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 15 mg / kg to about 100 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 30 mg / kg or 60 mg / kg per body weight of the subject once daily.

[0231] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 0.1 mg to about 100 mg once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 10 mg or about 50 mg once daily.

[0232] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 5 mg / kg to about 1000 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 10 mg / kg to about 550 mg / kg per body weight of the subject once daily. In some embodiments, non-naturally occurring melanocortin analog is administered at a dose of about 15 mg / kg to about 100 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 20 mg / kg to about 75 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 30 mg / kg or 60 mg / kg per body weight of the subject once daily.

[0233] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 5 mg / kg to at least 1000 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 10 mg / kg to at least 550 mg / kg per body weight of the subject once daily. In some embodiments, non-naturally occurring melanocortin analog is administered at a dose of at least 15 mg / kg to at least 100 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 20 mg / kg to at least 75 mg / kg per body weight of thesubject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 30 mg / kg or 60 mg / kg per body weight of the subject once daily.

[0234] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 5 mg / kg to at least about 1000 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 10 mg / kg to at least about 550 mg / kg per body weight of the subject once daily. In some embodiments, non-naturally occurring melanocortin analog is administered at a dose of at least about 15 mg / kg to at least about 100 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 20 mg / kg to at least about 75 mg / kg per body weight of the subject once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 30 mg / kg or 60 mg / kg per body weight of the subject once daily.

[0235] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 50 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 125 mg / kg, about 150 mg / kg, about 175 mg / kg, about 200 mg / kg, about 250 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, or about 500 mg / kg, about 550 mg / kg, about 600 mg / kg, about 650 mg / kg, about 700 mg / kg, about 750 mg / kg, about 800 mg / kg, about 850 mg / kg, about 900 mg / kg, about 950 mg / kg, or about 1000 mg / kg per body weight of the subject once daily.

[0236] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 5 mg / kg, at least 10 mg / kg, at least 15 mg / kg, at least 20 mg / kg, at least 25 mg / kg, at least 30 mg / kg, at least 35 mg / kg, at least 40 mg / kg, at least 50 mg / kg, at least 65 mg / kg, at least 70 mg / kg, at least 75 mg / kg, at least 80 mg / kg, at least 85 mg / kg, at least 90 mg / kg, at least 95 mg / kg, at least 100 mg / kg, at least 125 mg / kg, at least 150 mg / kg, at least 175 mg / kg, at least 200 mg / kg, at least 250 mg / kg, at least 300 mg / kg, at least 350 mg / kg, at least 400 mg / kg, at least 450 mg / kg, or at least 500 mg / kg, at least550 mg / kg, at least 600 mg / kg, at least 650 mg / kg, at least 700 mg / kg, at least 750 mg / kg, at least 800 mg / kg, at least 850 mg / kg, at least 900 mg / kg, at least 950 mg / kg, or at least 1000 mg / kg per body weight of the subject once daily.

[0237] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 5 mg / kg, at least about 10 mg / kg, at least about 15 mg / kg, at least about 20 mg / kg, at least about 25 mg / kg, at least about 30 mg / kg, at least about 35 mg / kg, at least about 40 mg / kg, at least about 50 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, at least about 100 mg / kg, at least about 125 mg / kg, at least about 150 mg / kg, at least about 175 mg / kg, at least about 200 mg / kg, at least about 250 mg / kg, at least about 300 mg / kg, at least about 350 mg / kg, at least about 400 mg / kg, at least about 450 mg / kg, or at least about 500 mg / kg, at least about 550 mg / kg, at least about 600 mg / kg, at least about 650 mg / kg, at least about 700 mg / kg, at least about 750 mg / kg, at least about 800 mg / kg, at least about 850 mg / kg, at least about 900 mg / kg, at least about 950 mg / kg, or at least about 1000 mg / kg per body weight of the subject once daily.

[0238] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 5 mg / kg per body weight of the subject once daily.

[0239] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 10 mg / kg per body weight of the subject once daily.

[0240] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 15 mg / kg per body weight of the subject once daily.

[0241] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 20 mg / kg per body weight of the subject once daily.

[0242] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 25 mg / kg per body weight of the subject once daily.

[0243] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 30 mg / kg per body weight of the subject once daily.

[0244] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 35 mg / kg per body weight of the subject once daily.

[0245] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 40 mg / kg per body weight of the subject once daily.

[0246] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 50 mg / kg per body weight of the subject once daily.

[0247] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 5 mg / kg per body weight of the subject once daily.

[0248] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 10 mg / kg per body weight of the subject once daily.

[0249] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 15 mg / kg per body weight of the subject once daily.

[0250] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 20 mg / kg per body weight of the subject once daily.

[0251] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 25 mg / kg per body weight of the subject once daily.

[0252] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 30 mg / kg per body weight of the subject once daily.

[0253] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 35 mg / kg per body weight of the subject once daily.

[0254] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 40 mg / kg per body weight of the subject once daily.

[0255] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least 50 mg / kg per body weight of the subject once daily.

[0256] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 5 mg / kg per body weight of the subject once daily.

[0257] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 10 mg / kg per body weight of the subject once daily.

[0258] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 15 mg / kg per body weight of the subject once daily.

[0259] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 20 mg / kg per body weight of the subject once daily.

[0260] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 25 mg / kg per body weight of the subject once daily.

[0261] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 30 mg / kg per body weight of the subject once daily.

[0262] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 35 mg / kg per body weight of the subject once daily.

[0263] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 40 mg / kg per body weight of the subject once daily.

[0264] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of at least about 50 mg / kg per body weight of the subject once daily.

[0265] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 50 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 125 mg / kg, about 150 mg / kg, about 175 mg / kg, about 200 mg / kg, about 250 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, about 500 mg / kg, about 550 mg / kg, about 600 mg / kg, about 650 mg / kg, about 700 mg / kg, about 750 mg / kg, about 800 mg / kg, about 850 mg / kg, about 900 mg / kg, about 950 mg / kg, or about 1000 mg / kg per body weight of the subject twice daily.

[0266] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 5 mg / kg per body weight of the subject twice daily.

[0267] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 10 mg / kg per body weight of the subject twice daily.

[0268] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 15 mg / kg per body weight of the subject twice daily.

[0269] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 20 mg / kg per body weight of the subject twice daily.

[0270] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 25 mg / kg per body weight of the subject twice daily.

[0271] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 30 mg / kg per body weight of the subject twice daily.

[0272] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 35 mg / kg per body weight of the subject twice daily.

[0273] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 40 mg / kg per body weight of the subject twice daily.

[0274] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 50 mg / kg per body weight of the subject twice daily.

[0275] In some embodiments, the non-naturally occurring melanocortin analog is administered at one or more doses. In some embodiments, the non-naturally occurring melanocortin analog is administered at two or more doses.

[0276] In some embodiments, the non-naturally occurring melanocortin analog is administered at a first dose once daily or twice daily. In some embodiments, the non- naturally occurring melanocortin analog is administered at a second dose once daily or twice daily.

[0277] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose that is a projected human equivalent dose (HED) based on a nonhuman primate dose. In some embodiments, the HED based on a 10 mg / kg nonhuman primate dose ranges from about 300 mg to about 550 mg.

[0278] In some embodiments, the non-naturally occurring melanocortin analog is administered at a first dose once daily for about 5 days to about 10 days. In some embodiments, the non-naturally occurring melanocortin analog is administered at a second dose once daily for about 5 days to about 10 days, after administration of the first dose. In some embodiments, the non-naturally occurring melanocortin analog is administered at a third dose twice daily for about 7 days to about 21 days, after administration of the second dose. In some embodiments, the second dose is greater than the first dose. In some embodiments, the third dose comprises a cumulative dose that is greater than the second dose.

[0279] In some embodiments, the non-naturally occurring melanocortin analogs or a pharmaceutical composition thereof (e.g., the pharmaceutical composition) is administered at a second dose that is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 250%, 300%, 350%, 400%, 450%, or 500% greater than the first dose.

[0280] In some embodiments, the non-naturally occurring melanocortin analogs or a pharmaceutical composition thereof (e.g., the pharmaceutical composition) is administered at a third dose that is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 250%, 300%, 350%, 400%, 450%, or 500% greater than the first dose and / or the second dose.

[0281] In some embodiments, the dosage of the second dose is greater than the dosage of the first dose. In some embodiments, the first dose is administered about once a day and the second dose is administered about twice a day.

[0282] In some embodiments, the dosage of the third dose is greater than the dosage of the first dose and / or the second dose. In some embodiments, the first dose and / or the second dose is administered about once a day, and the third dose is administered about twice a day.

[0283] In some embodiments, the non-naturally occurring melanocortin analog is administered using two or more different administration routes. The two or more different administration routes may comprise an oral administration and a subcutaneousadministration. The oral administration may occur before, during, or after the subcutaneous administration.

[0284] In some embodiments, the first dose and the second dose comprise different routes of administration. In some embodiments, the first dose comprises an oral dose and the second dose comprises a subcutaneous dose. In some embodiments, the first dose and / or second dose and the third dose comprise different routes of administration. In some embodiments, the first dose and / or the second dose comprises an oral dose and the third dose comprises a subcutaneous dose.

[0285] In some embodiments, the second dose is administered at least about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, or about 6 weeks after the first dose.

[0286] In some embodiments, the third dose is administered at least about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, or about 6 weeks after the first dose and / or the second dose.

[0287] In some embodiments, the non-naturally occurring melanocortin analog or a pharmaceutical composition thereof (e.g., the pharmaceutical composition) is administered at two or more dosage amounts, each dosage amount differing from the other. The first dosage amount, the second dosage amount, and the third dosage amount may each be greater than, less than, equal to, or substantially equivalent to another dosage amount. For example, the first dosage amount is less than a second dosage amount which is greater than a third dosage amount also greater than the first dosage amount. The dosage amounts refer to doses administered more than once over a period of time, such as a first dosage amount administered daily, BID, TID, or weekly, a second dosage amount administered daily, BID, TID, or weekly, and a third dosage amount administered daily, BID, TID, or weekly. In some embodiments, the first dosage amount, the second dosage amount, or the third dosage amount may be administered every other day, every second day, every third day, twice per week, or twice per month. For example, the first dosage amount of 10 mg / kg may be administered once a day for 7 days, followed by the second dosage amount of 20mg / kg once a day for 7 days, followed by the third dosage amount of 15 mg / kg twice a day for at least about 7 days. In some embodiments, the third dosage amount is administered for at least about 7 days, at least about 14 days, or at least about 21 days.

[0288] In some embodiments, the non-naturally occurring melanocortin analog or a pharmaceutical composition thereof (e.g., the pharmaceutical composition) may be administered using more than one route of administration. For example, the first dosage amount and the second dosage amount may be administered by two different routes of administration. In addition, the first dosage amount and the second dosage amount administered via different routes of administration may be the same or different dosage amounts. In some embodiments, the first dosage amount is administered orally and the second dosage amount is administered subcutaneously. The dosage frequency and / or duration of the dosage amounts may be the same or may be different. For example, the frequency of each dosage amount may be more than once per day, once per day, once every other day, once per week, or once every other week, or once per month. As another example, the duration of each dosage amount may be one day, two days, three days, more than three days, one week, more than one week, two weeks, more than two weeks, one month, or more than one month. For example, a dosing regimen may comprise a first dosage amount administered once a day or twice a day for about 5 weeks, and a second dosage amount administered subcutaneously once a day or twice a day for about 1 week. In some embodiments, the second dosage amount is administered the day after the duration of the first dosage amount is complete, or two days, three days, more than three days, one week, more than one week, two weeks, more than two weeks, one month, or more than one month after the first dosage amount is complete. In some embodiments, the second dosage amount is administered at least about 1 week after administration of the first dosage is complete.

[0289] In some embodiments, the non-naturally occurring melanocortin analog or a pharmaceutical composition thereof (e.g., the pharmaceutical composition) is administered at a dose of (i) 10 mg / kg once a day for 7 days, followed by (ii) 20 mg / kg once a day for 7 days, followed by (iii) 15 mg / kg twice a day for at least about 7 days. In some embodiments, (iii) is administered for at least about 7 days, at least about 14 days, or at least about 21 days.

[0290] In some embodiments, the non-naturally occurring melanocortin analog or a pharmaceutical composition thereof (e.g., the pharmaceutical composition) comprises a (i) first dose administered orally once a day or twice a day for about 5 weeks, and (ii) second dose administered subcutaneously once a day or twice a day for about 1 week. In some embodiments, the second dose is administered at least about 1 week after administration of the first dose is complete.

[0291] In some embodiments, the non-naturally occurring melanocortin analog is administered (i) at a first dose of about 5 mg / kg to about 20 mg / kg administered once a day for about 5 days to about 10 days; (ii) at a second dose of about 10 mg / kg to about 40 mg / kg administered once a day after administration of the first dose for about 5 days to about 10 days; (iii) at a third dose of about 5 mg / kg to about 30 mg / kg administered twice a day after administration of the second dose for at least about 5 days to about 10 days. In some embodiments, the third dose is administered twice a day for at least about 7 days or more.

[0292] In some embodiments, the non-naturally occurring melanocortin analog is administered (i) at a first dose of about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, or about 20 mg / kg administered once a day for about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days; (ii) at a second dose of about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, or about 40 mg / kg administered once a day after administration of the first dose about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days; (iii) at a third dose of about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, or about 30 mg / kg administered twice a day after administration of the second dose for at least about 5 days, about, 6 days, about 7 days, about 8 days, about 9 days, or about 10 days. In some embodiments, the third dose is administered twice a day for at least about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, or about 14 days or more.

[0293] In some embodiments, the non-naturally occurring melanocortin analog is administered (i) at a first dose of about 5 mg / kg administered once a day for about 7 days; (ii) at a second dose of about 15 mg / kg administered once a day after administration of the first dose for about 7 days; (iii) at a third dose of about 10 mg / kg administered twice a dayafter administration of the second dose for at least about 7 days. In some embodiments, the third dose is administered twice a day for at least about 14 days or more.

[0294] In some embodiments, the non-naturally occurring melanocortin analog is administered (i) at a first dose of about 10 mg / kg administered once a day for about 7 days; (ii) at a second dose of about 20 mg / kg administered once a day after administration of the first dose for about 7 days; (iii) at a third dose of about 15 mg / kg administered twice a day after administration of the second dose for at least about 7 days. In some embodiments, the third dose is administered twice a day for at least about 14 days or more.

[0295] In some embodiments, the non-naturally occurring melanocortin analog is administered (i) at a first dose of about 15 mg / kg administered once a day for about 7 days; (ii) at a second dose of about 25 mg / kg administered once a day after administration of the first dose for about 7 days; (iii) at a third dose of about 20 mg / kg administered twice a day after administration of the second dose for at least about 7 days. In some embodiments, the third dose is administered twice a day for at least about 14 days or more.

[0296] In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 0.1 mg to about 100 mg once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 0.5 mg to about 75 mg once daily. In some embodiments, the non-naturally occurring melanocortin analog is administered at a dose of about 5 mg to about 60 mg once daily. In some embodiments, wherein the non-naturally occurring melanocortin analog is administered at a dose of about 10 mg or about 50 mg once daily.

[0297] In some embodiments, the non-naturally occurring melanocortin analog is administered as a single dosage unit. In some embodiments, the non-naturally occurring melanocortin analog dose is administered as multiple dosage units. In some embodiments, the multiple dosage units individually comprise about 2.5 mg / mL to about 100 mg / mL of the non-naturally occurring melanocortin analog. In some embodiments, the multiple dosage units individually comprise about 2.5 mg / mL to about 10 mg / mL of the non-naturally occurring melanocortin analog. In some embodiments, the multiple dosage units individually comprise about 6 mg / mL of the non-naturally occurring melanocortin analog. In some embodiments, the non-naturally occurring melanocortin analog is administered as about 5dosage units to about 10 dosage units. In some embodiments, the non-naturally occurring melanocortin analog is administered as 5 dosage units. In some embodiments, the non- naturally occurring melanocortin analog is administered as 10 dosage units.

[0298] In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in a concentration of 5 mg / mL to 100 mg / mL, relative to a total volume of the pharmaceutical composition. In some embodiments, the non- naturally occurring melanocortin analog is present in the pharmaceutical composition in a concentration of about 10 mg / mL, about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 50 mg / mL, or about 60 mg / mL, relative to a total volume of the pharmaceutical composition. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition in a concentration of about 50 mg / mL, relative to a total volume of the pharmaceutical composition.

[0299] In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition at a dose of about 5 mg to about 1000 mg in a dose volume of about 1 mL to 20 mL, and wherein the administration is performed once, twice, 3 times, 4 times, 5 times, or 10 times per day. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition at a dose of about 50 mg to about 100 mg in a dose volume of about 2 mL to 10 mL, and wherein the administration is performed once, twice, 3 times, 4 times, 5 times, or 10 times per day. In some embodiments, the non-naturally occurring melanocortin analog is present in the pharmaceutical composition at a dose of about 75 mg in a dose volume of about 5 mL, and wherein the administration is performed once, twice, 3 times, 4 times, 5 times, or 10 times per day. In some embodiments, the administration is performed once per day.Methods

[0300] The present technology comprises methods of treating, preventing, reducing, or otherwise ameliorating one or more symptoms or conditions associated with the melanocortin system. In some embodiments, the methods treat, prevent, or otherwise reduce a disease, a condition, or a disorder associated with the melanocortin system or a symptom thereof. The melanocortin system of the present technology may compriseregulatory pathways exerted through melanocortin receptor signaling (e.g., activation or inhibition of a melanocortin receptor or a signal transduction therefrom).

[0301] In some embodiments, the present technology comprises treating, preventing, reducing, or otherwise ameliorating one or more symptoms, conditions, diseases, or disorders associated with melanocortin system dysfunction (e.g., dysfunction in energy balance regulation, appetite, and / or energy expenditure). Nonlimiting examples of such symptoms, conditions, diseases, or disorders include cachexia, lethargy, appetite, sleep, arousal, libido, locomotion, cardiovascular anomalies, vasodilatation, hypertension, hypotension, sodium regulation, pain, pain perception, homeostasis, endocrine and exocrine gland secretion, inflammation, addictive behavior, increasing endogenous opioid activity, or decreasing opioid tolerance.

[0302] In some embodiments, the present technology is useful in treating, preventing, mitigating, or reducing symptoms of a condition or disease associated with dysfunction of the gut-brain axis. In some embodiments, the present technology is useful in treating, preventing, mitigating, or reducing symptoms of irritable bowel syndrome.

[0303] In some embodiments, the present technology is useful in treating, preventing, reducing, or otherwise ameliorating bodily conditions such as weight regulation (e.g., obesity, anorexia, and cachexia), hormonal secretion (e.g., dry eye and / or dry mouth syndrome), immuno-relevant conditions, or sexual dysfunction.

[0304] In some embodiments, the subject of the present technology is administered a non-naturally occurring melanocortin analog at least about 30 seconds, 1 minute, 5 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 12 hours, 24 hours, 36 hours, 2 days, 3 days, 4 days, 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 15 weeks, 20 weeks, 30 weeks, 40 weeks, 1 year, or 2 years after diagnosis or identification of the disease, condition, disorder, or the symptom thereof.

[0305] In some embodiments, the methods comprise administering a non-naturally occurring melanocortin analog at a first dose and a second to a subject in need thereof, wherein the second dose is greater than the first dose. In some embodiments, the subject is administered a third dose that is greater than the first and the second dose.

[0306] In some embodiments, the methods comprise administered the non-naturally occurring melanocortin analog to the subject in increase doses or increased dosages, relative to the first dose.Maintenance Doses

[0307] In some embodiments, a maintenance dose of the non-naturally occurring melanocortin agonist is administered to the subject to maintain or enhance one or more outcomes of the methods of the present technology.Subjects

[0308] In some embodiments, the subject is a mammal, including but not limited to a human, a non-human primate such as a chimpanzee, a domestic livestock or a farm animal such as a cow, a bison, sheep, a pig, a goat, a horse, a chicken, and a rooster, a domestic pet animal such as a dog, a cat, a rat, a mouse, and a rabbit, and a laboratory subject such as a rodent, including a rat, a mouse, and a guinea pig. In some embodiments, the subject is a human. In some embodiments, the subject is an animal such as a rat or a dog.Controls

[0309] The controls of the present technology may comprise the subject at baseline. In some embodiments, the controls of the methods of the present technology comprise a subject that is not administered a non-naturally occurring melanocortin analog of the present technology or a subject subjected to a method lacking one or more steps of the methods of the present technology.Pharmacokinetics and Pharmacodynamics

[0310] The non-naturally occurring melanocortin analogs of the present technology exhibit pharmacokinetic (pK) and / or pharmacodynamic (pD) parameters. Such pK and / or pD may be expressed or otherwise determined relative to a control, which, in some instances, may be a non-naturally occurring melanocortin analog lacking one or more features of the non-naturally occurring melanocortin analogs of the present technology.

[0311] In some embodiments, the pK and / or pD of the non-naturally occurring melanocortin analogs may be assessed using concentration and / or temporal measurements(e.g., Tfinai, Cmax, T (h)), AUC, or Tmax In some embodiments, the non-naturally occurring melanocortin analogs have reduced clearance and / or metabolism, increased uptake, absorption, and / or stability, relative to a control.Clearance

[0312] “Clearance” may refer to the elimination, absorption, and / or metabolism of the non-naturally occurring melanocortin analogs in the subject’s plasma. Clearance may be assessed as volume of plasma cleared of the non-naturally occurring melanocortin analogs over time (e.g., mL / min, L / hr, or L / day) and / or may be normalized to body weight of the subject (e.g., mL / min / kg). Reduced clearance may also be represented by an increase in half-life or volume of distribution (Vd). In some embodiments, measuring clearance comprises measuring a terminal elimination rate constant (Az) or an inter-compartmental clearance (Q).

[0313] In some embodiments, the reduction in clearance comprises a measurement about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 day, 6 days, or 1 week after administration of the non-naturally occurring melanocortin analogs. In some embodiments, the reduced clearance comprises an intestinal fluid clearance, a gastric fluid clearance, a liver clearance, or a liver microsome clearance.

[0314] In some embodiments, the reduction in clearance comprises a measurement during administration of the non-naturally occurring melanocortin analogs. In some embodiments, the reduction in clearance comprises a measurement at the completion of administration of the non-naturally occurring melanocortin analogs.Concentration

[0315] In some embodiments, the non-naturally occurring melanocortin analogs of the present technology comprise an increased tissue, plasma, serum, or aqueous concentration relative to a control. “Concentration” may comprise a measurement reflecting one or more of the absolute amounts of the non-naturally occurring melanocortin analogs, the absorption of the non-naturally occurring melanocortin analogs, the metabolism of non-naturally occurring melanocortin analogs, or the elimination of non-naturally occurring melanocortinanalogs. The aqueous concentration may comprise an intestinal fluid concentration or a gastric fluid concentration.

[0316] The increased tissue, plasma, and / or serum concentration may be an increase in concentration of the non-naturally occurring melanocortin analogs at a given time point relative to a control administered at the same dose and measured at the same time point. The concentration may be measured at an intermediate time point or a final time point and may be measured as a mean residence time (MRT), an average concentration (Cavg), a trough concentration (Ctrough), or a concentration at the end of administration (e.g., infusion) time (CT).

[0317] In some embodiments, the increased concentration is reflected by an increase in peak plasma concentration (Cmax). An increase in Cmax may suggest increased absorption, reduced metabolism, or slower elimination of the non-naturally occurring melanocortin analogs, relative to a control. In some embodiments, Cmax comprises a dose normalized Cmax (DNCmax).

[0318] In some embodiments, the increased concentration is reflected by an increase in minimum plasma concentration (Cmin). An increase in Cmin may suggest increased absorption, reduced metabolism, or slower elimination of the non-naturally occurring melanocortin analogs, relative to a control. In some embodiments, Cmin comprises a dose normalized Cmin (DNCmin).

[0319] In some embodiments, the increased concentration is reflected by a reduction in time to reach Cmax (Tmax). A reduced Tmax may suggest increased absorption, reduced metabolism, or slower elimination of the non-naturally occurring melanocortin analogs, relative to a control.

[0320] In some embodiments, the increased concentration is reflected by a final measurable concentration (Tfinai). An increased Tfinai may suggest increased absorption, reduced metabolism, or slower elimination of the non-naturally occurring melanocortin analogs, relative to a control.

[0321] In some embodiments, the increased concentration is reflected by an increase in area under the curve (AUC). An increase in AUC may signify increased exposure to thenon-naturally occurring melanocortin analogs and / or may suggest increased absorption, reduced metabolism, or slower elimination of the non-naturally occurring melanocortin analogs, relative to a control. The AUC measurement may comprise an Area Under the Curve for Concentration of Drug in Non-Compartmental Analysis (DNAUC).

[0322] In some embodiments, the increased concentration is reflected by a reduction in a partition coefficient or an increase in a partition coefficient, relative to a control. The reduced partition coefficient may reflect an increase in aqueous solubility (e.g., an intestinal fluid or a gastric fluid), relative to the control. In some embodiments, the increased partition coefficient may reflect an increase in membrane permeability, relative to the control.

[0323] In some embodiments, the increase in tissue, plasma, and / or serum concentration comprises a measurement about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1 .5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 day, 6 days, or 1 week after administration of the non- naturally occurring melanocortin analogs.

[0324] In some embodiments, the increase in tissue, plasma, and / or serum concentration comprises a measurement at least 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1 .5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 day, 6 days, or 1 week after administration of the non- naturally occurring melanocortin analogs.

[0325] In some embodiments, the increase in tissue, plasma, and / or serum concentration comprises a measurement at least about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 day, 6 days, or 1 week after administration of the non-naturally occurring melanocortin analogs.

[0326] In some embodiments, the increase in tissue, plasma, and / or serum concentration comprises a measurement during administration of the non-naturally occurring melanocortin analogs.

[0327] In some embodiments, the increase in tissue, plasma, and / or serum concentration comprises a measurement at the completion of administration of the non- naturally occurring melanocortin analogs.

[0328] In some embodiments, the concentration of the non-naturally occurring melanocortin analog in the plasma or a tissue of the subject is at least about 5 ng / mL to at least about 2000 ng / mL about 24 hours after administration of the non-naturally occurring melanocortin analog.

[0329] In some embodiments, the non-naturally occurring melanocortin analog is administered to the subject until the concentration of the non-naturally occurring melanocortin analog in the plasma or a tissue of the subject is at least about 5 ng / mL to at least about 2000 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 5 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 10 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 50 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 100 ng / mL after administration of the non- naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 200 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 300 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 400 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 500 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the wherein concentration is at least about 750 ng / mL after administration of the non- naturally occurring melanocortin analog. In some embodiments, the concentration is at least about 1000 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the is at least about 1500 ng / mL after administration of the non- naturally occurring melanocortin analog. In some embodiments, the concentration is at leastabout 2000 ng / mL after administration of the non-naturally occurring melanocortin analog. In some embodiments, the concentration is a Cmax.Distribution

[0330] In some embodiments, the non-naturally occurring melanocortin analogs of the present technology comprise an increased distribution relative to a control. The increased distribution may be an increase in distribution of the non-naturally occurring melanocortin analogs at a given time point relative to a control administered at the same dose and measured at the same time point. The distribution may be measured at an intermediate time point or a final time point.

[0331] In some embodiments, the measurement of distribution comprises measuring a volume of distribution at the terminal phase (Vd or Vdii), a central volume of distribution (V), a peripheral volume of distribution (V2), an apparent volume of distribution (Vz), or a measurement of distribution comprises measuring a volume of distribution at steady state (Vss). A high or increased Vd, Vdu, Vz, and / or Vssmay suggest large distribution beyond the tissue, plasma, and / or serum compartment, relative to the control.

[0332] In some embodiments, the increase in distribution comprises a measurement about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 day, 6 days, or 1 week after administration of the non-naturally occurring melanocortin analogs.

[0333] In some embodiments, the increase in distribution comprises a measurement at least 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 day, 6 days, or 1 week after administration of the non-naturally occurring melanocortin analogs.

[0334] In some embodiments, the increase in distribution comprises a measurement at least about 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 day, 6 days, or 1 week after administration of the non-naturally occurring melanocortin analogs.

[0335] In some embodiments, the increase in distribution comprises a measurement during administration of the non-naturally occurring melanocortin analogs.

[0336] In some embodiments, the increase in distribution comprises a measurement at the completion of administration of the non-naturally occurring melanocortin analogs.Additional pD and pK Embodiments

[0337] In some embodiments, the non-naturally occurring melanocortin analog of the present technology exhibits one or more of the following:(a) an increase in half-life relative to a control;(b) a reduction in clearance relative to a control;(c) an increase in tissue concentration relative to a control;(d) an increase in plasma concentration relative to a control;(e) an increase in serum concentration relative to a control;(f) an increase in distribution relative to a control;(g) an increase in an AUC measurement relative to a control;(h) an increase in a DNALIC measurement relative to a control;(i) an increase in Tfinai relative to a control;(j) an increase in Cmax relative to a control;(k) an increase in Cmin relative to a control;(l) an increase in DNCmin relative to a control;(m) an increase in MRT relative to a control;(n) an increase in Cavg relative to a control;(o) an increase in Ctrough relative to a control;(p) an increase in CT relative to a control;(q) an increase in Vd relative to a control;(r) a reduction in Tmax relative to a control; or(s) a reduction in Q relative to a control.

[0338] In some embodiments, the non-naturally occurring melanocortin analog of the present technology exhibits one or more of the following, relative to a control:(a) an increase in half-life by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(b) a reduction in clearance by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control;(c) an increase in tissue concentration by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(d) an increase in plasma concentration by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(e) an increase in serum concentration by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(f) an increase in distribution by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(g) an increase in an AUC by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% measurement relative to a control;(h) an increase in a DNAUC measurement by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(i) an increase in Ttinai by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(j) an increase in Cmax by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(k) an increase in Cmin by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(l) an increase in DNCmin by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(m) an increase in MRT by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(n) an increase in Cavg by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(o) an increase in Ctrough by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(p) an increase in CT by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(q) an increase in Vd by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1000% relative to a control;(r) a reduction in Tmax by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control; or(s) a reduction in Q by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to a control.EXAMPLES

[0339] 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

[0340] The non-naturally occurring melanocortin analogs of the present technology 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.

[0341] 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.

[0342] 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(1980)). An exemplary solid-phase synthesis of non-naturally occurring melanocortin analogs is provided below.

[0343] Initially, the C-terminal amino acid residue of the non-naturally occurring melanocortin analog was coupled to a solid-phase support, e.g., a solid-phase resin. 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-chlorotrity I chloride resin via an ester linkage. Some methods of coupling rely on the formation of an aminde 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.

[0344] 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. 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. 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

[0345] During synthesis of the non-naturally occurring melanocortin analogs, reactive side chain groups of the various amino acid residues were protected with suitable protectinggroups, which prevented undesirable chemical reaction from occurring at that site until the protecting group was removed.

[0346] 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- m ethoxybenzyloxycarbonyl (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.

[0347] Alpha aminoprotecting 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

[0348] 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.

[0349] 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 (TBTU), 2-(1 H- benzotriazol-1 -yl)-1 ,1 ,3,3-tetramethyluronium hexafluorophosphate (HBTU), 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 (TPTU), N,N'-dicyclohexylcarbodiimide / 1 - hydroxybenzotriazole (DCCI / HOBt)) 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

[0350] The agonist and antagonist activity of exemplary non-naturally occurring melanocortin analogs at the melanocortin receptors (e.g., MC1 R, MC3R, MC4R, and MC5R) were 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

[0351] Compounds were 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 solvent volume defined. 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. Each 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.Table 1. Cell linesCompound Testing

[0352] Compounds were 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-0232C) 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

[0353] Cyclic AMP (cAMP) Homogenous Time-Resolved Fluorescence (HTRF) assay for Gs coupled receptor:

[0354] 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 1 mM IBMX or 25pM Rolipram).

[0355] Dose response curves were performed in parallel with the reference compounds.

[0356] 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.

[0357] For antagonist test (384well): 5 l 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

[0358] On each day of experimentation and prior to the testing of compounds, reference compounds were tested at several concentrations in duplicate (n=2) to obtain a dose-response curve and an estimated EC50 and / or IC50 values.

[0359] Reference values thus obtained for the test were compared to historical values obtained from the same receptor and used to validate the experimental session.

[0360] 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.Non-naturallv Occurring Melanocortin Analog Grouping

[0361] Group A included non-naturally occurring melanocortin analogs A1 to A10, all of which are cyclic peptides comprising the motif Pro-dPhe-Arg-Trp (SEQ ID NO: 122), or a derivative thereof. Derivatives of the motif having SEQ ID NO: 122 may include substitution of Pro for another amino acid, for example, dlle, dVal, His, or dAla, or deletion of Pro. Group A non-naturally occurring melanocortin analogs are provided in Table 2.Table 2. Group A non-naturally occurring melanocortin analogs

[0362] Group B included non-naturally occurring melanocortin analogs B1 to B2, all of which are cyclic peptides comprising the derivatives of the motif Pro-dPhe-Arg-Trp (SEQ ID NO: 122). Derivatives of the motif having a sequence of SEQ ID NO: 122 include insertion of a residue, for example His, between Pro and dPhe. Group B non-naturally occurring melanocortin analogs are provided in Table 3.Table 3. Group B non-naturally occurring melanocortin analogs

[0363] Group C included non-naturally occurring melanocortin analogs C1 to C6, all of which are cyclic peptides comprising the motif Trp-Pro-dPhe-Arg-Trp (SEQ ID NO: 120) or a derivative thereof. Derivatives of the motif having SEQ ID NO: 120 may include substitution of Pro for another amino acid, for example, dLeu or dVel, or substitution of Trp for another amino acid, for example, dNal(1 ’). Group C non-naturally occurring melanocortin analogs are provided in Table 4.Table 4. Group C non-naturally occurring melanocortin analogs

[0364] Group D included non-naturally occurring melanocortin analogs D1 to D, all of which are cyclic peptides comprising the motif Ala-dNal(2')-Arg-Trp or a derivative thereof. Derivatives of the motif Ala-dNal(2')-Arg-Trp may include substitution of Ala for another amino acid, for example, Leu, dLey, Vai, or Gly. Group D non-naturally occurring melanocortin analogs are provided in Table 5.Table 5. Group D non-naturally occurring melanocortin analogs

[0365] Group E included non-naturally occurring melanocortin analogs D1 to D, all of which are cyclic peptides comprising the motif Arg-p(l)dPhe-Arg-Tic or a derivative thereof. Derivatives of the motif Ala Arg-p(l)dPhe-Arg-Tic may include substitution of p(l)dPhe for another amino acid, for example, dBip. Group E non-naturally occurring melanocortin analogs are provided in Table 6.Table 6. Group E non-naturally occurring melanocortin analogs

[0366] Group F included non-naturally occurring melanocortin analogs F1 , F3, F4 and F6-F8, all of which are cyclic peptides comprising the motif dTrp-p(CI)dPhe-Arg-Trp (SEQ ID NO: 121 ) or a derivative thereof. Derivatives of the motif having SEQ ID NO: 121 may include substitution of dTrp for another amino acid, for example, dGIn, dPhe, or dBip, or substitution of p(CI)dPhe for another amino acid, for example, p(F)dPhe. Group D non- naturally occurring melanocortin analogs are provided in Table 7.Table 7. Group F non-naturally occurring melanocortin analogsAgonist activity of melanocortin analogs on melanocortin 1 and 5 receptors

[0367] Administration of some non-naturally occurring melanocortin analogs activated melanocortin 1 receptor (MC1 R) and / or melanocortin 5 receptor (MC5R) activity, as measured by cAMP levels (Table 8).Table 8. Dose-response results of melanocortin analogs and control against the melanocortin 1 receptor (MC1 R) and melanocortin 5 receptor (MC5R)n.c. = not calculatedAntagonist activity of melanocortin analogs on melanocortin 3 and 5 receptors

[0368] Administration of some non-naturally occurring melanocortin analogs inhibited melanocortin 3 receptor (MC3R) and / or melanocortin 5 receptor (MC4R) activity, as measured by cAMP levels.Table 9. Dose-response results of melanocortin analogs and control against the melanocortin 3 receptor (MC3R) and melanocortin 5 receptor (MC4R)n.c. = not calculated n.t. = not testedExample 5: Pharmacokinetic Assessment of Non-Naturally Occurring Melanocortin Analogs Following Oral Administration to Rats and Cynomolgus MonkeysStudy Objective

[0369] The objective of this study is to determine the pharmacokinetics of non-naturally occurring melanocortin analogs of the present technology following oral gavage administration in male rats and single oral administration to non-naive male cynomolgus monkeys. In rats, the test article will be monitored in plasma for up to 24 hours post each dose. Study design and sample collection will be conducted as outlined in Table 10 and Table 11.Table 10. Study DesignTable 11. Sample CollectionsE: Anticoagulant: Potassium (K2) EDTA;P: plasmaC: cerebrospinal fluid‘Sampled only for certain compounds

[0370] Dose Solution Analysis Samples: After each dose preparation, remove approximately 0.5 mL aliquots from the formulations, transfer the aliquots into amber HPLC vials and stored at -60°C or lower until assayed in duplicate for dose validation.

[0371] Disposition of Remaining Test Article Formulations: Remaining formulations will be stored at -60°C or lower.

[0372] Disposition of Remaining Test Article (dry powder or solid): Remaining test article will be stored at room temperature desiccated, and protected from light until shipment or discard.

[0373] Animals will be fasted overnight through 4 hours post dosing.Vehicle and Formulation Preparation:

[0374] Appropriate amount of test article will be accurately weighed and mixed with appropriate volume of vehicle to get a clear solution or suspension.

[0375] Formulation samples will be removed from each of the formulation solutions, transferred into 1.5 mL of polypropylene microcentrifuge tubes and run dose validation by LC / UV or LC-MS / MS.

[0376] Compounds to be assessed are outlined in Table 12.Table 12. CompoundsCF: FW / MW / purityAnimal Specifications: Cynomolgus Monkeys

[0377] Cynomolgus Monkey specifications are outlined in Table 13.Table 13. Cynomolgus Monkey Specifications

[0378] Environmental Conditions: Environment controls will be set to maintain a temperature range of 20-26°C, a relative humidity range of 40 to 70%, and a 12-hour light / 12-hour dark cycle. The light / dark cycle may be interrupted for study-related activities.

[0379] Housing: Animals will be group-housed (up to four animals / sex / cage) in polysulfone cages with certified aspen shaving bedding or corncob bedding during acclimation and study period. While animals may be individually housed after surgery orwhen there is special requirement in protocol, as well as, for behavioral or health reasons or due to cage mate death.

[0380] Diet and Feeding: Animals were offered certified rodent breeding and growth diet ad libitum every day, unless fasted for study procedures. Each lot of the diet is analyzed for nutrients, chemical contaminant and microorganisms, the results are reviewed and evaluated by veterinarians before provided to animals.

[0381] Drinking Water: Autoclaved RO (reverses osmosis) water will be available to all animals, ad libum.

[0382] Feed and Water Analyses: Autoclaved RO water will be provided ad libitum via water bottle. Water samples are periodically analyzed by a certified laboratory for specified microorganisms and environment contaminants. The diet is routinely analyzed by the manufacturer for specified microorganisms, nutritional components and environmental contaminants.

[0383] Environmental Enrichment: Enrichment toys will be provided.

[0384] Dose Administration: The dose volume will be determined by the animals' body weight collected on the morning of dosing day.Animal Specifications: Rats

[0385] Rat specifications are outlined in Table 14.Table 14. Rat SpecificationsObservations and Examinations

[0386] Clinical Observations: All animals will be observed at dosing and each scheduled collection. All abnormalities will be recorded.

[0387] Body Weight: All animals will be weighed on the dosing day prior to dosing to determine the dose volume to be administered.Sample Collection and Processing

[0388] Blood Sample Collection and Process: At least 0.1 mL blood will be collected at each time point. All blood samples will be collected via jugular vein. All blood samples will be transferred into low binding EP tube with anticoagulant (0.5 M Potassium (K2) EDTA will be pre-added as a ratio of 50:1 for blood: anticoagulant), 0.05% Triton X-100 (e.g., 100uL Blood+2uL 2.5% Triton X-100) will be used for desorption the blood samples will be placed on wet ice.

[0389] Blood samples will be centrifuged within 1 hr of collection at 3,200 g 4°C for 10 minutes. Following centrifugation, plasma samples will be transferred into their respective pre-labeled low binding EP tube and immediately frozen over dry ice. The plasma samples will be stored lower than -60°C until bioanalysis.

[0390] LC-MS / MS method development:

[0391] A LC-MS / MS method for the quantitative determination of test compound in biological matrix will be developed.

[0392] N in 1 cassette LC-MS / MS method may be developed for samples coming from different studies as long as these studies belong to the same sponsor.

[0393] Cassette administration assay could be performed if the mass difference (AMass) among different analytes is >4 Da. In this case, interference evaluation is not necessary.

[0394] If AMass among different analytes is less than 4 Da, there is a potential risk that interference would occur during LC-MS / MS analysis. If such kind of cassette assay is still requested by client, interference among analytes will not be evaluated but the LC separation of those analytes by using a generic method will be attempted.

[0395] Sample analysis:

[0396] A calibration curve with at least 6 non-zero calibration standards will be applied for each batch including LLOQ.

[0397] If sample number within a batch is < 12, at least one set of standard curve separated with two parts through begin and end of the sequence should be included in the run and QCs are not required. The recommended injection order is C8, C6, C4, C2, study samples, C7, C5, C3, C1.

[0398] If sample number within a batch is > 12, one standard curve and two sets of QCs with low, middle and high concentrations will be applied for bioanalysis. Meanwhile, QCs number should be more than 5% of study sample number.

[0399] Samples, coming from one client with the same type of matrix in different studies, are allowed to be quantified in one analysis run by using the developed N in 1 cassette LC-MS / MS method.

[0400] Acceptance criteria:

[0401] (1 ) Linearity: At least 75% of the calibration standards should fall within ±20% of their nominal values for biofluid and within ±25% of their nominal values for tissue and feces samples. If the endpoints, such as LLOQ and ULOQ, on the calibration curve are eliminated, the calibration curve will be truncated. The truncated calibration curve should consist of at least 75% of the initial STDs.

[0402] (2) Accuracy: At least 67% of QCs should fall within ±20% of their nominal values for biofluid and within ±25% of their nominal values for tissue and feces samples. At least half of QCs at each concentration should be passed.

[0403] (3) Specificity: The mass response of analyte in the double blank and blank should be < 50% LLOQ.

[0404] (4) Sensitivity: The LLOQ will be tried to target < 3 ng / mL. Any adjustment ofLLOQ will be informed to client in advance.

[0405] (5) Carryover: The mean calculated carryover peak area in the double blanks or blanks immediately after the highest standard injection should be less than that of LLOQ. If the carryover couldn’t meet the criteria, the impact of the carryover on unknown samples should be re-evaluated according to the below procedure:

[0406] Carryover should be re-evaluated based on absolute carryover. Absolute carryover is calculated by carryover contribution multiplies carryover impact, where the carryover contribution is calculated by the area ratio of the double blank or blank with the highest carryover (Area max of carryover blank) to the ULOQ with the minimum calculated value (Area min of ULOQ), and the carryover impact is calculated by the area ratio of one injection (Area of one injection) to the following injection (Area of the following injection). The absolute carryover should be below the acceptable accuracy of the studies (e.g., 20% or 25%).

[0407] Carryover contribution = Areamax of carryover blank / Areamin of ULOQ Carryover impact = Area of one injection / Area of the following injection Absolute carryover = Carryover contribution * Carryover impactData Analysis

[0408] Plasma concentration versus time data for Compounds A-C in Rats and Cynomolgus Monkeys will be plotted in graph and analyzed by non-compartmental approaches. Related PK parameters will be calculated according to dosing route, e.g., Cl, Vdss and CO for intravenous administration, Cmax, Tmax or %F for extravascular administration, and T1 / 2, AUC(o-t), AUC(o -inf), MRT(o-t), MRT(o-inf) for all routes. Preliminary plasma results after24 hours for 60 mg / kg oral administration in Rats and 30mg / kg of Compounds A-C in Cynomolgus Monkeys are shown in Table 15 and FIG. 1.Table 15. Plasma Pharmacokinetic Results of Compounds A-C after 24 hours

[0409] Plasma concentration versus time data for Compounds D (07) (FIGS. 2A-2C) and E (010) (FIGS. 2D-2I) in Cynomolgus Monkeys was plotted in graphs and analyzed by non-compartmental approaches. Related PK parameters were calculated according to dosing route, e.g., Cl, Vdss and CO for intravenous administration, Cmax, Tmax or %F for extravascular administration, and , AUC(o-t), AUC(o-int), MRT(o-t), MRT(o-inf) for all routes. Preliminary plasma results after 24 hours of 3 mg / kg, 10 mg / kg, 30 mg / kg, or 60 mg / kg PO administration of Compound D (Ac-Nle-c[Glu-Pro-p(F)dPhe-Arg-Trp-Om]-dVal-dPro-NH2; 07) and Compound E (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Orn]-dVal-dPro-NH2; 010) in Cynomolgus Monkeys are shown in Table 16.Table 16. Plasma Pharmacokinetic Results of Compounds D (07) and E (010) in Cynomolgus Monkeys after 24 hours

[0410] Preliminary plasma and cerebrospinal fluid (CSF) results after 24 hours following oral administration of Compound D (Ac-Nle-c[Glu-Pro-p(F)dPhe-Arg-Trp-Orn]- dVal-dPro-NH2; 07), Compound E (Ac-Nle-c[Glu-His-p(F)dPhe-Arg-Trp-Om]-dVal-dPro- NH2; 010), and Compound F (Ac-Nle-c[Asp-Pro-dPhe-Arg-Trp-Lys]-dVal-dPro-NH2) in Rats are shown in Table 17 and Table 18. Graphs of plasma concentration and CSF concentration in Rats are shown in FIGS. 3A-3F (PO administration).

[0411] Plasma and CSF concentrations over 24 hours after oral administration of Compound D (07), Compound E (010), or setmelanotide were also assessed (FIGS. 4H- 4K). It was predicted that 010 will be efficacious for treating general obesity in part due to the PK values in plasma and CSF and the higher affinity for MC3R and MC4R relative to setmelanotide.Table 17. Plasma Pharmacokinetic Results of Compounds D (07) and E (010) in Rats after 24 hoursTable 18. CSF Pharmacokinetic Results of Compounds D (07) and E (010) in Rats after24 hoursND = Not determined

[0412] Dose proportionality of Compound D (07) and Compound E (010) following a single administration via PO in Rats is shown in Table 19.Table 19. Dose Proportionality of Compounds D and E in Rats after a single PO administrationPlasma Concentration

[0413] Plasma pharmacokinetic measurements using 30 mg / kg were assessed in Cynomolgus monkey plasma up to 24 hours. Results are detailed in Table 20.Table 20: Pharmacokinetic Data

[0414] The antagonist activity of exemplary non-naturally occurring melanocortin analogs at specific ion channels was measured using the Qube electrophysiological platform. The non-naturally occurring melanocortin analogs and specific ion channel targets are provided in Table 21 .Table 21. Ion channel targets and melanocortin analogsMethods

[0415] The non-naturally occurring melanocortin analogs identified above were tested for antagonist activity at various ion channels at concentrations ranging from 0.1 mM to 30 mM. In each experiment and if applicable, the respective reference compounds were tested concurrently with the test compounds, and the data were compared with known historical values. hNayl .5 Sodium Channel Assay - Qube APC

[0416] Onset and steady state block of peak Navi .5 current is measured using a pulse pattern, repeated every 5 sec, consisting of a hyperpolarizing pulse to -120mV for a 200ms duration, depolarization to -15mV amplitude for a 40ms duration, followed by step to 40mVfor 200ms and finally a 100ms ramp (1 .2 V / s) to a holding potential of -80mV. Peak current is measured during the step to -15mV.

[0417] The parameters measured were difference between the peak inward current on stepping to -15mV (i.e. , peak of the current) and the leak current. All data were filtered for seal quality, seal drop, and current amplitude. The peak current amplitude was calculated before and after compound addition and the amount of block was assessed by dividing the Test compound current amplitude by the Control current amplitude. Control is the mean Nav1.5 current amplitude collected 10 seconds at the end of the vehicle control; Test Compound is the mean Nav1.5 current amplitude collected 10 seconds at the end of test concentration application for each concentration. hKv4.3 / hKChlP2 Potassium Channel Assay - Qube APC

[0418] After whole cell configuration is achieved, the cells are held at -80m V. Onset and steady state block of hKv4.3 current is measured using a pulse pattern from -80mV to 40mV amplitude for a 110ms duration, and finally a 100ms ramp (1.2 V / s) to -80m V. This paradigm is delivered once every 5s to monitor the current amplitude.

[0419] The parameters measured were the maximum outward current evoked on stepping to 40mV from holding potential of -80mV. All data were filtered for seal quality, seal drop, and current amplitude. The peak current amplitude was calculated before and after compound addition and the amount of block was assessed by dividing the Test compound current amplitude by the Control current amplitude. Control data is the mean Kv4.3 / KChlP2 current amplitude collected 10 seconds at the end of the vehicle control period; Test compound data is the mean Kv4.3 / KChlP2 current amplitude collected 10 seconds at the end of test concentration application for each concentration. hCayl .2 (L-tvoe) CiPA Calcium Channel Assay - Qube APC

[0420] Onset and steady state block of peak hCavl .2 current is measured using a pulse pattern, repeated every 15 sec. Cells were held at -80mV for a 50ms before stepping to -90mV for 100ms to measure leak current and then stepped back to -80mV for 50ms, depolarization to OmV amplitude for a 40ms duration, followed by step to 40mV for 200msand finally a 100ms ramp (1.2 V / s) to a holding potential of -80 mV. Peak current is measured during the step to OmV. Each concentration is applied for 5 minutes.

[0421] The calcium current amplitude is calculated by measuring the difference between the peak inward current on stepping to OmV or the peak inward current at the ramp (i.e. peak of the current) and the leak current. The calcium current is assessed in vehicle control conditions and at the end of each five (5) minute compound application. hNayl .5 Late Current Sodium Channel Assay - Qube APC

[0422] Onset and steady state block of Late Navi .5 current is measured using a pulse pattern, repeated every 5 sec, consisting of a hyperpolarizing pulse to -120mV for a 200ms duration, depolarization to -15mV amplitude for a 40ms duration, followed by step to 40mV for 200ms and finally a 100ms ramp (1.2 V / s) to a holding potential of -80mV. Late current is measured as charge current elicited during the ramp with 50nM ATXII.

[0423] The parameters measured were the ramp current charge (AUC) evoked on ramping back to -80mV from 40mV test pulse in the presence of 50nM ATXII. All data were filtered for seal quality, seal drop, and current. The peak and ramp current amplitude was calculated before and after compound addition and the amount of current was assessed by dividing the Test compound current by the Control current. Control data is the mean hNavl .5 late current collected 15 seconds at the end of 50nM ATXII application (50nM ATXII control); Test compound data is the mean ramp hNavl .5 current collected 15 seconds at the end of test concentration application for each concentration. hERG Potassium Channel Assay - Qube APC

[0424] After whole cell configuration is achieved, the cells are held at -80mV. Cells are held at this voltage for 50ms to measure the leak current, which is subtracted from the tail current on-line. The cells are depolarized to +40mV for 500ms and then to -80 mV over a 100ms ramp to elicit the hERG tail current. This paradigm is delivered once every 8s to monitor the current amplitude. All compounds were tested in the presence of 0.1 % Pluronic F-68 Non-lonic Surfactant and at approximately room temperature.

[0425] The parameters measured were the maximum tail current evoked ramping back to -80mV from the test pulse of 40mV. All data were filtered for seal quality, seal drop, andcurrent amplitude. The peak current amplitude was calculated before and after compound addition and the amount of block was assessed by dividing the Test compound current amplitude by the Control current amplitude. Control data is the mean hERG current amplitude collected for three pulses (24 seconds) at the end of the vehicle control; Test compound data is the mean hERG current amplitude collected for three pulses (24 seconds) at the end of test concentration application for each concentration. hKCNQ1 / hminK Potassium Channel Assay - Qube APC

[0426] After whole cell configuration is achieved, the cells are held at -80mV. KCNQ1 / minK currents are evoked by a 1000ms pulse from -80mV to 60mV followed by a ramp from 60mV to -80mV over 115ms with the outward peak currents measured upon depolarization of the cell membrane. This paradigm is delivered once every 15s to monitor the current amplitude.

[0427] The parameters measured were the maximum outward current evoked on stepping to +60mV from a holding potential of -80mV. All data were filtered for seal quality, seal drop, and current amplitude. The peak current amplitude was calculated before and after compound addition and the amount of block was assessed by dividing the Test compound current amplitude by the Control current amplitude. Control data is the mean hKCNQ1 / hminK current amplitude collected 30 seconds at the end of vehicle control period; Test compound data is the mean hKCNQ1 / hminK current amplitude collected 30 seconds at the end of test concentration application for each concentration. hKir2.1 Potassium Channel Assay - Qube APC

[0428] After whole cell configuration is achieved, the cells are held at -30mV. Kir2.1 currents are evoked by a single 500ms pulse to -120mV before returning to the holding potential of -30mV. This paradigm is delivered once every 20s to monitor the current amplitude.

[0429] The parameters measured were the maximum inward current elicited on stepping to -120mV for 500ms from a holding potential of -30 mV. All data were filtered for seal quality, seal drop, and current amplitude. The peak current amplitude was calculated before and after compound addition. Residual non-Kir2.1 current was eliminated vianormalization to residual current after application of 100uM Barium Chloride. The amount of Test compound effect was then assessed by dividing the Test compound current amplitude by the Control current amplitude. Control data is the mean Kir2.1 current amplitude collected 40 seconds at the end of the vehicle control; Test compound data is the mean Kir2.1 current amplitude collected 30 seconds at the end of test concentration application for each concentration.Results

[0430] Where presented, ICso values were determined by a non-linear, least squares regression analysis. Reference standards were run as an integral part of each assay to ensure the validity of the results obtained. Results from the ion channel assessment of non- naturally occurring melanocortin analogs are provided in Table 22.Table 22. Dose-response results of melanocortin analogs and reference compounds against ion channels.*N / C: Indicates observed mean maximal inhibition was <25%Example 7: Assessment of physical biological properties of non-naturally occurring melanocortin analogs

[0431] Physical biological properties including solubility, in vitro absorption and in vitro metabolism of nine exemplary non-naturally occurring melanocortin analogs were assessed according to the following procedures. In each experiment and if applicable, the respective reference compound was tested concurrently with the test compounds, and the data were compared with known values. Stock solutions of the tested compounds were prepared at a concentration of 0.01 M in DMSO.Solution Properties

[0432] Solution properties of non-naturally occurring melanocortin analogs and reference compounds in various biological media were assessed according to the conditions provided in Table 23.Table 23. Assay conditions for in vitro absorption assessmentAqueous Solubility

[0433] Aqueous solubility (pM) was determined by comparing the peak area of the principal peak in a calibration standard (200 pM) containing organic solvent (methanol / water, 60 / 40, v / v) with the peak area of the corresponding peak in a buffer sample. In addition, chromatographic purity (%) was defined as the peak area of the principal peak relative to the total integrated peak area in the HPLC chromatogram of the calibration standard. A chromatogram of the calibration standard of each test compound, along with a UV / VIS spectrum with labeled absorbance maxima, was generated.

[0434] A chromatogram of the test compound (200 pM) along with a UV / VIS spectrum with labeled absorbance maxima, was generated.Protein Binding

[0435] The peak areas of the test compound in the buffer and test samples were used to calculate percent binding and recovery according to the following formulas:Protein binding(%) = ((AreaP-Areab) / AreaP) x 100Recovery(%) = ((AreaP-Areab) / Areac) x 100 where: AreaP= peak area of analyte in protein matrix; Areab = peak area of analyte in buffer; and Areac= peak area of analyte in control sample.Partition Coefficient

[0436] The total amount of compound was determined as the peak area of the principal peak in a calibration standard (100 pM) containing organic solvent (methanol / water, 60 / 40, v / v). The amount of compound in buffer was determined as the combined, volume-corrected, and weighted areas of the corresponding peaks in the aqueous phases of three organic- aqueous samples of different composition. An automated weighting system was used toensure the preferred use of raw data from those samples with well quantifiable peak signals. The amount of compound in organic was calculated by subtraction. Subsequently, Log D was calculated as the Log10 of the amount of compound in the organic phase divided by the amount of compound in the aqueous phase.Half-Life Determination

[0437] At the end of the incubation at each of the time points, an equal volume of an organic mixture (acetonitrile / methanol, 50 / 50, v / v) was added to the incubation mixture. Samples were analyzed by HPLC-MS / MS and corresponding peak areas were recorded for each analyte. The ratio of precursor compound remaining after each time point relative to the amount present at time 0, expressed as percent, is reported as chemical stability. The half-life (T1 / 2) was estimated from the slope of the initial linear range of the logarithmic curve of compound remaining (%) versus time, assuming first order kinetics.

[0438] Results of the solubility assessment detailed above are provided in Tables 24- 33.Table 24. Protein binding of melanocortin analogs in plasmaTable 25. Half-life of melanocortin analogs in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF)Table 26. Aqueous solubility of melanocortin analogs in plasma (PBS), simulated gastric fluid (SGF), and simulated intestinal fluid (SIF)*pH = 7.4Table 27. Partition coefficient of melanocortin analogs and reference compounds*log D, n-octanol / PBS, pH 7.4In Vitro Absorption

[0439] In vitro absorption of non-naturally occurring melanocortin analogs was determined using permeability assays. Assay conditions are provided in Table 28.Table 28. Assay conditions for in vitro absorption assessmentPermeability

[0440] The apparent permeability coefficient (Papp) of the test compound was calculated as follows:

[0441] Papp(cm / s) = (VR*CR,end / Dt) X (1 / A*(CD,mid_CR,mid)

[0442] where VR is the volume of the receiver chamber; CR.end is the concentration of the test compound in the receiver chamber at the end time point; At is the incubation time; A is the surface area of the cell monolayer; CD, mid is the calculated mid-point concentration of the test compound in the donor side, which is the mean value of the donor concentration at time 0 minute and the donor concentration at the end time point; and CR.mid is the midpoint concentration of the test compound in the receiver side, which is one half of the receiver concentration at the end time point. Concentrations of the test compound were expressed as peak areas of the test compound.Recovery of the Test Compound from the Permeability Assay

[0443] The recovery of the test compound was calculated as follows:

[0444] Recovery(%) = ((VD*CD,end+VR*CR,end) / D*CDo) x 100

[0445] where VD and VR are the volumes of the donor and receiver chambers, respectively; CD, end is the concentration of the test compound in the donor sample at the end time point; CR.end is the concentration of the test compound in the receiver sample at the end time point; and CDO is the concentration of the test compound in the donor sample at time zero. Concentrations of the test compound are expressed as peak areas of the test compound.Fluorescein assessment for Permeability assays

[0446] Fluorescein was used as the cell monolayer integrity marker. Fluorescein permeability assessment (in the A-B direction at pH 7.4 on both sides) was performed after the permeability assay for the test compound. The cell monolayer that had a fluorescein permeability of less than 1 .5 x 10'6cm / s for Caco-2 and MDR1 -MDCKII cells and 2.5 x 10'6cm / s for MDCKII cells was considered intact, and the permeability result of the test compound from intact cell monolayer is reported.

[0447] Results of the in vitro absorption assessments described above are provided in Table 29.Table 29. In vitro absorption of melanocortin analogs and reference compoundsIn Vitro Metabolism

[0448] In vitro metabolism of non-naturally occurring melanocortin analogs was determined using the assay conditions provided in Table 30.Table 30. Assay conditions for in vitro metabolism assessment

[0449] Metabolic stability, expressed as percent of the parent compound remaining, was calculated by comparing the peak area of the compound at the time point relative to that at time-0. The half-life (T1 / 2) was estimated from the slope of the initial linear range of the logarithmic curve of compound remaining (%) vs. time, assuming the first-order kinetics. The apparent intrinsic clearance (CLmt, in pL / min / pmol, pL / min / mg or pL / min / Mcell) was calculated according to the following formula:CLint= 0.693 / (Ti / 2*(mg protein / pL or million cells / pL or pmol CYP isoyme / pL)).

[0450] Results from the intrinsic clearance assay are provided in Table 31 .Table 31. In vitro metabolism of melanocortin analogs and reference compoundsPhysiochemical and ADME Properties

[0451] Physiochemical and Absorption, Distribution, Metabolism, and Excretion (ADME) properties of 010 were assessed (Tables 32 and 33). 010 may be efficiently manufactured using standard solid-phase peptide synthesis (SPPS). B07, a melanocortin antagonist, has high structural homology to 010. B07 has been made in GMP batches up to 1 kg and it is amenable to efficient SPPS with high overall yields. Non-canonical residues and cyclic structure may restricts fermentation-based approaches, and peptide purification is performed using standard RP-HPLC techniques.

[0452] 010 may be amenable to liquid-phase peptide synthesis (LPPS) for late-stage clinical and commercial manufacturing, offering potential for significant reductions in manufacturing cost. It is anticipated that typical scale-up cost reductions (e.g., use of bulk residues, process optimization, captive manufacturing cost setups) will emerge.Table 32: 010 Physicochemical PropertiesTable 33: O10 ADME Properties* SGF: simulated gastric fluid** SIF: simulated intestinal fluidExample 8: CYP Inhibition of Non-Naturally Occurring Melanocortin Analogs

[0453] The inhibitory potential of exemplary non-naturally occurring melanocortin analogs was tested on seven human Cytochrome P450 (CYP) enzymes: CYP1A, CYP2B6, CYP2C19, CYPC8, CYP2C9, CYP2D6, and CYP3A. Exemplary melanocortin analogs 07 and 010 and comparator melanocortin analog B07 (Ac-Nle-c[Asp-Pro-dNal(2’)-Arg-Trp- Lys]-dVal-dPro-NH2; SEQ ID NO: 95) were tested at concentrations ranging from 0.1 mM to 100 mM for inhibition of the seven CYPs. The CYP inhibition assays were performed using human liver microsomes (HLM) and human recombinant CYP isozymes in 96-well plate format. Direct inhibition (e.g., zero-min incubation) and time-dependent (e.g., 30 min preincubation) inhibition assays were performed. The results of the CYP inhibition assays of 07 and 010 at 10 mM are shown in Table 34 and the results of the comparator CYP inhibition assays including ICso values at each of the CYP enzymes are shown in Table 35.Table 34. CYP Inhibition Studies of 010Table 35. CYP Inhibition of TCMCB07Example 9: Caloric Intake, Body Weight, and Body CompositionFood Intake and Body Composition in Monkeys

[0454] Changes in caloric intake and body weight were assessed in diet-induced obese Cynomolgus monkeys (n=6). Monkeys were orally administered 10 mg / kg 07 (MC4R selective agonist) or O10 (MC3R / MC4R coagonist) once daily. One day 15, dosing changed to 15 / mg / kg administered twice daily, and dosing was maintained until at least day 28 andmonkeys were assessed until at least day 35. Control monkeys were administered saline. Daily caloric intake (FIGS. 4A), cumulative caloric intake (FIGS. 4B and 5A), percent change in caloric consumption, relative to baseline (FIGS. 4C, 5B), and caloric intake per body weight were measured in addition to food intake (FIGS. 15A-5R; Tables 36-54). Cumulative caloric intake and percent change in caloric intake, relative to baseline, were each reduced in the monkeys administered 07 or 010, relative to those administered saline.

[0455] Food preference towards a lower-fat diet in the monkeys administered 010 was also observed, where 010-treated monkeys consumed 50% more reduced-fat feed (FIG. 5N) and 20% less high-fat feed (FIG. 50) on average compared to monkeys administered saline.

[0456] Body composition was assessed, including body weight (kg), body height (m), BMI (kg / m2), whole body bone mineral content (g), trunk bone mineral content (g), bone density (g / cm2), trunk fat mass (g), trunk fat mass (%), trunk muscle mass (g), trunk muscle mass (%), fat mass (g), fat mass (%), muscle mass (g), muscle mass (%), and lean mass (%) (Tables 55-60) (FIGS. 7A-7N). Diet-induced obese monkeys administered 010 showed a 7.5% weight loss in 5 weeks, relative to baseline.Table 36: Animal Pre-screening and Day 1 Food IntakeTable 37: Days 2-4 Food IntakeTable 38: Days 5-7 Food IntakeTable 39: Days 8-10 Food IntakeTable 40: Days 11-13 Food IntakeTable 41 : Day 14 Food IntakeTable 42: Days 15 and 16 Food IntakeTable 43: Days 17 and 18 Food IntakeTable 44: Days 19 and 20 Food IntakeTable 45: Days 21 and 22 Food IntakeTable 46: Days 24 and 25 Food IntakeTable 47: Days 26 and 27 Food IntakeTable 48: Days 28 and 29 Food IntakeTable 49: Day 30 Food IntakeTable 50: Food Intake (Calorie) Prescreening and Days 1-4Table 51: Days 5-10 Food Intake (Calorie)Table 52: Days 11 -14 Food Intake (Calorie)Table 53: Days 15-22 Food Intake (Calorie)Table 54: Days 24-30 Food Intake (Calorie)Table 55: Body Composition at Baseline (Part 1 )Table 56: Body Composition at Baseline (Part 2)Table 57: Body Composition Day 29 (Part 1)Table 58: Body Composition Day 29 (Part 2)Table 59: Body Composition Change (Part 1)Table 60: Body Composition Change (Part 2)

[0457] By day 8, the body weight change relative to baseline was most reduced in the monkeys administered 010, compared to monkeys administered saline (FIGS. 6A-6M) (Tables 61 and 62). Monkeys were administered 010 orally for 36 days and abstained from dosing until day 43. From days 43-50, monkeys were subcutaneously administered 010, with dosing discontinued after day 50 (FIG. 6I). A decrease in percent change in body weight was observed in monkeys administered 010, relative to saline. Following dosing discontinuation, body weight was maintained and did not rebound during the day 50-71 observations (FIG.6J).Table 61 : Body Weight (kg) (Day 1 -22)Table 62: Body Weight (Percent Change)

[0458] Blood chemistry, clinical chemistry, gastric impact, and nausea levels were also assessed (FIGS. 8A-8D; Tables 63-74). Blood levels chemistry levels were unchanged by day 29 from baseline in monkeys orally administered 010 relative to those administered saline. Cage side observations showed no signs of gastric distress (assessed by stool changes) or nausea (assessed by food refusal).Table 63: Clinical Chemistry Day 1 (Part A)Table 64: Clinical Chemistry Day 1 (Part B)Table 65: Clinical Chemistry Day 15 (Part A)Table 66: Clinical Chemistry Day 15 (Part B)Table 67: Clinical Chemistry Day 29 (Part A)Table 68: Clinical Chemistry Day 29 (Part B)Table 69: Complete Blood Count Day 1 (Part A)Table 70: Complete Blood Count Day 1 (Part B)Table 71: Complete Blood Count Day 15 (Part A)Table 72: Complete Blood Count Day 15 (Part B)Table 73: Complete Blood Count Day 29 (Part A)Table 74: Complete Blood Count Day 29 (Part B)Pigmentation Observations

[0459] Changes in pigmentation were observed in the monkeys orally administered 010, where, at high doses of 010, a tanning effect was observed. The tanning effect comprised skin darkening without hyperpigmentation (e.g., an uneven distribution of melanin) compared to baseline. Notably, the tanning effect was only seen in typically sun- exposed skin, with no observation of ectopic tanning (e.g., palms, gums, etc.). Monkeys were never exposed to sunlight. Pigmentation resolved back to baseline levels rapidly after treatment was discontinued.

[0460] The slope of the 010 weight loss curve (FIG. 6I, 6M, 6K) suggests that a lower dose of 010 may be employed for the same efficacy. This suggests presence of a therapeutic window between weight loss and pigmentation (i.e., tanning effect). Combinations with weight loss agents may permit usage of lower doses of both agents, limiting side effects, and increasing therapeutic window further.Food Intake and Body Composition in Rats

[0461] Normalized cumulative food consumption (FIG. 5Q), and percent change in body weight (FIG. 6L) were assessed in rats administered MC4R selective agonists (A07D, 07, 011 ) (n=5) or MC3R / MC4R co-agonists (010) compared to saline controls (n=4). For each treatment group, rats were subcutaneously administered 0.5 mg / kg of A07D, 07, 010, or 011 for days 1 -7 and 1 mg / kg for days 8-17. 011 and 07 did not appear to influence food consumption as a percent of food consumption in saline-treated rats, relative to 010 (FIG. 5Q). At 17 days, 010 treated rats gained only about 0.64% of their body weight compared to saline, which gained about 8.98% body weight (**) (FIG. 16L). 010-treated rats gained less weight than 07-treated rats (****), A07D-treated rats (*), and 011 -treated rats (p=.O66). 010, an MC3R / MC4R coagonist, also exhibited a stronger influence on caloric consumption, as measures by cumulative food consumption) than the MC4R selective agonists. However, both the MC4R selective agonists and the MC3R / MC4R coagonists were each more effective in reducing body weight relative to saline controls.Example 10: Cardiac Effects07, 010, & 011

[0462] Changes in cardiac effects were assessed in rats subcutaneously administered 07, 010, or 011 at either 0.5 mg / kg, 1 mg / kg, or 3 mg / kg. Control rats were administered saline. Animal specifications are outlined in Table 75.Table 75: Animal SpecificationsAnimal Housing

[0463] 6-7 week old male SD rats were obtained and housed in a controlled environment (target conditions: temperature 20 to 24°C, relative humidity 30 to 70%). Temperature and relative humidity was monitored daily. An electronic time-controlled lighting system was used to provide a 12-hour light / 12-hour dark cycle. Rats were housed in plastic cages. Enrichment toys were provided. During the housing and study period, the rats were fed with normal chow and fresh water, ad libitum.Animal Acclimation

[0464] After arrival, the animals acclimated to environment for at least 5 days.Study Design

[0465] Before the experiment, the weight of each animal was measured and rats were be separated into different groups based on weight, with 3 rats per group (Table 76).Table 76: Study GroupsBlood Pressure and 2-Lead ECG Monitoring

[0466] After acclimatization, animals were anesthetized with urethane as well as subjected to femoral vein for drug injection and carotid common artery to measure blood pressure. Meanwhile, electrocardiogram (ECG) was detected by subcutaneous puncture needle electrodes. The body temperature of the rats was maintained at about 37°C during assessments and experimentation. The blood pressure (BP) and ECG was recorded for 15 minutes before administration and for 3 hours after administration continuously for each rat. Readouts

[0467] Systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR) were recorded with blood pressure measurements.

[0468] QRS, ST, QT, P wave, T wave, and HR were assessed with ECG measurements.

[0469] Recording duration occurred from -15 min (pre-dose) to 180 min (after dosing).Blood Collection

[0470] After recording, animals were euthanized with CO2 immediately and blood was collected for in vitro analysis, including plasma corticosterone by ELISA Kit.Data Processing and Analysis

[0471] All values were expressed as mean ± SEM. The significances of the differences among groups and within groups was evaluated by one-way AN OVA followed by Dunnett’s test using Graph Pad statistic software. A p value of less than 0.05 was considered statistically significant. Results for the BP analysis are shown for all groups in Tables 77-79 and for groups 1 and 5-7 in FIGS. 8A-8D. ECG results are shown in Tables 80-84.Table 77: Blood Pressure Analysis (Part 1 )Table 78: Blood Pressure Analysis (Part 2)GroupTable 79: Blood Pressure Analysis (Part 3)Table 80: ECG Results (Part 1 )Table 81 : ECG Results (Part 2)Table 82: ECG Results (Part 3)Table 83: ECG Results (Part 4)Table 84: ECG Results (Part 5)

[0472] No significant changes in systolic blood pressure (SBP) (FIG. 9A), diastolic blood pressure (DBP) (FIG. 9B), nor heart rate (HR) (FIG. 9C) were observed in rats administered 010 at any dose, relative to controls. Comparably, setmelanotide-treated animals showed at some doses, there were changes in QTc, suggesting altered cardiac repolarization. (FIGS. 10A-10D).

[0473] Normalized heart rate (HR), QTc, systolic blood pressure (SBP), diastolic blood pressure (DBP), mean blood pressure (MBP), QT interval, QRS interval, and RR interval were also assessed in male cynomolgus monkeys orally administered 3 mg / kg, 10 mg / kg, 30 mg / kg, or 60 mg / kg 010 (FIGS. 11A-11 H). Relative to control cynomolgus monkeys administered saline, no significant cardiac effects were observed in monkeys administered 010 at any dose. In contrast, monkeys administered setmelanotide experienced an increase in SBP, DBP, and MBP, relative to monkeys administered saline. This increase was also observed during the dark cycle (“lights off” FIGS. 11 C-11 E). No increase in these parameters were observed at any time point in monkeys administered 010, relative to those administered salineExample 11: Bioavailability of Non-Naturally Occurring Melanocortin Analogs

[0474] Various pharmacokinetic parameters were assessed in the plasma of male cynomolgus monkeys fasted prior to dosing A07D at 10 mg / kg or 30 mg / kg orally. Results are shown in Table 85.Table 85: A07D Results

[0475] ND = Not determined (Parameters not determined due to inadequately defined terminal elimination phase).

[0476] BQL = Below the lower limit of quantitation (LLOQ).

[0477] If the adjusted rsq (linear regression coefficient of the concentration value on the terminal phase) is less than 0.9, T1 / 2 might not be accurately estimated.

[0478] If the % AUCExtra > 20%, AUCo-inf, Cl, MRTo-inf and Vdss might not be accurately estimated.

[0479] If the % AUMCExtra > 20%, MRTo-inf and Vdssmight not be accurately estimated.

[0480] The adjusted linear regression coefficient of the concentration value on the terminal phase is less than 0.9, T1 / 2 might not be accurately estimated.

[0481] a: Bioavailability (%) was calculated using AUCo-inf (if all AUCExtra < 20%) or AUCo-iast (if one or more AUCExtra > 20%) with Nominal Dose.

[0482] Plasma concentration of 07 and L2 were next assessed up to 24 hours following oral administration to cynomolgus monkeys at 10 mg / kg or 30 mg / kg, results of which are shown in FIGS. 14 and 16. 010 plasma concentration up to 24 hours following oral administration to cynomolgus monkeys at 3 mg / kg, 10 mg / kg, 30 mg / kg, or 60 mg / kg was also assessed, as shown in FIG. 15.Example 12: Assessing Off-Target Binding of O1Q

[0483] O10 was assessed for interaction with RFamide receptors, which are hypothesized to cause cardiac activation in MC4R-targeting small molecules and peptides (Table 86).Table 86: 010 Assessment for RFamide Receptor Binding

[0484] Generally, 010 did not agonize or antagonize RFamide receptors up to 1000 nM (top concentration tested), 1000x-100,000x above their respective EC50s on MC4R. Additionally, no hERG inhibition observed from 010 at concentrations up to 30 pM.

[0485] 010 was next screened for binding against a panel of 87 receptors and channels at 10 pM. Results showing an inhibition or stimulation higher than 50% were considered to represent significant effects of the test compounds. O10 only showed >50% inhibition of control on melanocortin receptors (FIG. 12). 010 was shown to have minimal inhibition at 10 pM for cytochrome P450 (CYP) enzymes (Table 87 and Table 88). 010 showed no meaningful impact on CYP induction or inhibition at physiologically relevant concentrations, demonstrating minimal drug-drug interactions.Table 87: Assessing Time Dependent CYP InhibitionTable 88: Assessing 010 and CYP Induction*Positive controls: Omeprazole (50 pM) for CYP1A2; Phenobarbital (1 mM) for CYP2B6; Rifampicin (10 pM) for CYP3A4.Example 13: Assessing Adipose Tissue Effects of O1Q

[0486] To assess adipose tissue effects from 010 administration, fat tissue was harvested from diet-induced obese cynomolgus monkeys orally administered 10 mg / kg 010or saline control. 010-treated monkeys showed (1 ) smaller lipid droplets, (2) darker, dense staining likely attributed to higher mitochondrial concentrations, and (3) increased vascularization, relative to saline controls (FIG. 13; H&E staining, 200x), consistent with beige / brown adipocytes. This suggests that 010 administration may stimulate metabolic remodeling through adipose tissue browning, which in turn may increase metabolic rate and calorie burn and may improve insulin sensitivity.Example 14: Assessing TCMCB07 BioavailabilityOral Bioavailability

[0487] To assess the oral bioavailability of TCMCB07 (Ac-Nle-c[Asp-Pro-dNal(2’)-Arg- Trp-Lys]-dVal-dPro-NH2; SEQ ID NO: 95), TCMCB07 was subjected to in vitro assays to evaluate absorption and stability. TCMCB07 levels were first assessed for stability in human plasma, simulated gastric fluid (SGF), and simulated intestinal fluid (SIF) over a 120-minute incubation period (FIG. 17).

[0488] TCMCB07 plasma concentration was next measured following oral administration in cynomolgus monkeys at 30.0 mg / kg (FIG. 18), the results of which are shown in Table 89. TCMCB07 plasma concentration in male and female-divided cynomolgus monkeys at the same concentration or oral administration are shown in Table 90.Table 89: Pharmacokinetics of TCMCB07 (ng / mL)ND = Not determinedBQL = Below the lower limit of quantitation (LLOQ)Table 90: TCMCB07 Plasma Concentration in Male and Female MonkeysSubcutaneous Administration

[0489] TCMCB07 pharmacokinetics were next assessed following 10 mg / kg SC administration in male Sprague Dawley rats (Table 91 ). This showed that TCMCB07 was detectable in the plasma for at least 24 hours following SC administration.Table 91 : Plasma TCMCB07 levels in Rodents

[0490] TCMCB07 plasma concentration was further assessed in rats and Beagle dogs following IP or oral administration (Table 92). This showed that TCMCB07 was detectable in the plasma across species tested and through multiple administration routes.Table 92: TCMCB07 Administration in Different Species - Plasma LevelsAdditional Embodiments

[0491] Various embodiments of the present technology are set forth below in paragraphs

[0493] to

[0496] :

[0492] Embodiment 1. A non-naturally occurring melanocortin analog comprising a sequence of Formula (IA(iii)):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA(iii)), wherein:X1is Nle;R1is Asp or Glu;R2is Pro;R3is His;R4is dPhe;R5is Arg;R6is Trp;R7is Lys or Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp or Glu at R1and Lys or Orn at R7

[0493] Embodiment 2. The non-naturally occurring melanocortin analog of embodiment 1 , wherein the sequence of Formula (IA(iii)) comprises a sequence of SEQ ID NO: 35 or 37.

[0494] Embodiment 3. The non-naturally occurring melanocortin analog of embodiment 1 or 2, wherein the sequence of Formula (IA(iii)) comprises a sequence of SEQ ID NO: 35.

[0495] Embodiment 4. The non-naturally occurring melanocortin analog of embodiment 1 or 2, wherein the sequence of Formula (IA(iii)) comprises a sequence of SEQ ID NO: 37.

[0496] From the foregoing, it will be appreciated that specific embodiments of the present technology have been described herein for purposes of illustration, but that various modifications may be made without deviating from the scope of the present technology. Accordingly, the present technology is not limited except as by the appended claims.

Claims

CLAIMSI / We claim:

1. A non-naturally occurring melanocortin analog comprises a sequence of Formula (IA(iii)):X1-R1-R2-R3-R4-R5-R6-R7-Y1-Y2(IA(iii)), wherein:X1is Nle;R1is Asp or Glu;R2is Pro;R3is His;R4is dPhe;R5is Arg;R6is Trp;R7is Lys or Orn;Y1is dVal;Y2is dPro; and the non-naturally occurring melanocortin analog is cyclized through a lactam bridge between Asp or Glu at R1and Lys or Orn at R7.

2. The non-naturally occurring melanocortin analog of claim 1 , wherein the sequence of Formula (IA(iii)) comprises a sequence of SEQ ID NO: 35 or 37.

3. The non-naturally occurring melanocortin analog of claim 1 or 2, wherein the sequence of Formula (IA(iii)) comprises a sequence of SEQ ID NO: 35.

4. The non-naturally occurring melanocortin analog of claim 1 or 2, wherein the sequence of Formula (IA(iii)) comprises a sequence of SEQ ID NO: 37.

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