MC4r agonist peptides and methods of use thereof

Abstract

Provided herein are melanocortin 4 receptor (MC4R) agonist peptides and methods of use thereof for the treatment and / or prevention of eating disorders (e.g., overeating), metabolic disorders (e.g., disorders resulting in positive energy imbalance), emotional / mental disorders, and / or dietary or syndromic obesity. In particular, provided herein are MC4R agonist peptides that exhibit enhanced selectivity for MC4R over other melanocortin receptors (e.g., MC1R, MC3R, MC5R). The peptides herein may exhibit enhanced in vitro potency, in vivo efficacy, pharmacokinetic properties, and / or stability compared to other known melanocortin 4 receptor binding peptides.

Description

[0001] Atorney Docket No. UM-43572.601

[0002] Client Ref. No. UM 2025-153 MC4R AGONIST PEPTIDES AND METHODS OF USE THEREOF

[0003] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0004] This invention was made with government support under DK127817 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0005] CROSS-REFERENCE

[0006] This application claims the benefit of U.S. Provisional Patent Application No.

[0007] 63 / 754,711, filed February 6, 2025, U.S. Provisional Patent Application No. 63 / 757,844, filed February 13, 2025, and U.S. Provisional Patent Application No. 63 / 782,434, filed April 2, 2025, each of which is incorporated by reference herein in its entirety.

[0008] SEQUENCE LISTING STATEMENT

[0009] The content of the electronic sequence listing titled UM_43572_601_SequenceListing.xml (Size: 217,690 bytes; and Date of Creation: February 6, 2026) is herein incorporated by reference in its entirety.

[0010] FIELD

[0011] Provided herein are melanocortin 4 receptor (MC4R) agonist peptides and methods of use thereof for the treatment and / or prevention of eating disorders (e.g., overeating), metabolic disorders (e.g., disorders resulting in positive energy imbalance), emotional / mental disorders, and / or dietary or syndromic obesity. In particular, provided herein are MC4R agonist peptides that exhibit enhanced selectivity for MC4R over other melanocortin receptors (e.g., MC1R, MC3R, MC5R). The peptides herein may exhibit enhanced in vitro potency, in vivo efficacy, pharmacokinetic properties, and / or stability compared to other known melanocortin 4 receptor binding peptides.

[0012] BACKGROUND

[0013] Obesity is a multifactorial condition involving an excessive amount of body fat.

[0014] Obesity is a medical problem that increases the risk of other diseases and health problems, such as heart disease, diabetes, high blood pressure and certain cancers. A large body of research has established the critical role of hypothalamic AgRP neural circuits in stimulating feeding and intense effort has focused on identifying pharmacological targets that suppressAtorney Docket No. UM-43572.601

[0015] Client Ref. No. UM 2025-153 these circuits as potential therapeutics for obesity. Therapeutics for the treatment / prevention of dietary or syndromic obesity, the upstream causes, and downstream effects are needed.

[0016] SUMMARY

[0017] Provided herein are melanocortin 4 receptor (MC4R) agonist peptides and methods of use thereof for the treatment and / or prevention of eating disorders (e.g., overeating), metabolic disorders (e.g., disorders resulting in positive energy imbalance), emotional / mental disorders, and / or dietary or syndromic obesity. In particular, provided herein are MC4R agonist peptides that exhibit enhanced selectivity for MC4R over other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). The peptides herein may exhibit enhanced in vitro potency, in vivo efficacy, pharmacokinetic properties, and / or stability compared to other known melanocortin 4 receptor binding peptides.

[0018] In some embodiments, peptide herein exhibits selectivity for MC4R over other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). In some embodiments, a peptide herein exhibits 2-fold or greater selectivity for MC4R over MC3R (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or more or ranges therebetween). In some embodiments, an MC4R agonist peptide herein exhibits 2-fold or greater selectivity for MC4R over MC1R (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or more or ranges therebetween). In some embodiments, an MC4R agonist peptide herein exhibits 2-fold or greater selectivity for MC4R over MC2R (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or more or ranges therebetween). In some embodiments, an MC4R agonist peptide herein exhibits 2-fold or greater selectivity for MC4R over MC5R (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or more or ranges therebetween).

[0019] In some embodiments, provided herein are methods, e.g., of treating an eating disorder (e.g., overeating), a metabolic disorder (e.g., disorders resulting in positive energy imbalance), emotional / mental disorders, and / or obesity in a subject, the methods comprising administering a melanocortin 4 receptor (MC4R) agonist to a subject suffering from the eating disorder. In some embodiments, the eating disorder is characterized by overeating. In some embodiments, the eating disorder is characterized by one or more emotional / mentalAtorney Docket No. UM-43572.601

[0020] Client Ref. No. UM 2025-153 symptoms. In some embodiments, the eating disorder is characterized by anxiety and / or depression. In some embodiments, the eating disorder is stress-induced overeating. In some embodiments, the MC4R agonist is a peptide. In some embodiments, the administration is repeated on a recurring basis for a period of at least 1 week (e.g., 1 week, 2 weeks, 1 month, 2 months, 4 months, 6 months, 9 months, 1 year, 2 years, 3, years, 4 years, or more). In some embodiments, the administration is repeated on a daily basis. In some embodiments, the administration is repeated on a twice-daily basis. In some embodiments, the administration is repeated on alternate days. In some embodiments, the administration is repeated on a weekly basis. In some embodiments, the administration is repeated on a recurring basis for a period of at least 1 month (e.g., 1 month, 2 months, 4 months, 6 months, 9 months, 1 year, 2 years, 3, years, 4 years, or more). In some embodiments, the administration is repeated on a recurring basis for a period of at least 1 year.

[0021] In some embodiments, provided herein are compositions comprising a peptide having 4 or fewer (e.g., 4, 3, 2, 1, 0) substitutions relative to SEQ ID NO: 1; wherein AA1 is linked to AA2 if AA1B is absent; wherein the peptide comprises at least one of: bAla at AAO; Phe, Hph, Gly, Ala at AA1B; Pro, D-Nle, D-Hle, D-Alb, or beta-Ala at AA3; Agp, Nar, Cit, or Alb at AA4; D-Fpy, D-Fpt, or D-Fpm at AA5; Wpb at AA7; or Abu, Hie, Cpa, Phg, D-Hph, or Om at AA9. In some embodiments, the peptide comprises 100% sequence similarity to SEQ ID NO: 1. In some embodiments, the peptide comprises SEQ ID NO: 1.

[0022] In some embodiments, provided herein are compositions comprising a peptide having 4 or fewer (e.g., 4, 3, 2, 1, 0) substitutions relative to SEQ ID NO: 2; wherein AA1 is linked to AA2 if AA1B is absent; wherein the peptide comprises at least one of: bAla at AAO; Phe, Hph, Gly, Ala at AA1B; Pro, D-Nle, D-Hle, D-Alb, or beta-Ala at AA3; Agp, Nar, Cit, or Alb at AA4; D-Fpy, D-Fpt, or D-Fpm at AA5; Wpb at AA7; or Abu, Hie, Cpa, Phg, D-Hph, or Om at AA9. In some embodiments, the peptide comprises 100% sequence similarity to SEQ ID NO: 2. In some embodiments, the peptide comprises SEQ ID NO: 2.

[0023] In some embodiments, AA4 is Alb. In some embodiments, AA4 is Cit. In some embodiments, AA7 is Wpb.

[0024] In some embodiments, the peptide has 4 or fewer (e.g., 4, 3, 2, 1, 0) substitutions relative to one of SEQ ID NOS: 3-22, 42-71, 81-86, and / or 95-111. In some embodiments, the peptide has 100% sequence similarity with one of SEQ ID NOS: 3-22, 42-71, 81-86, and / or 95-111. In some embodiments, the peptide comprises the sequence of one of SEQ ID NOS: 3-22, 42-71, 81-86, and / or 95-111. In some embodiments, the peptide furtherAtorney Docket No. UM-43572.601

[0025] Client Ref. No. UM 2025-153 comprises an X and / or Y cap as defined herein (e.g., N-terminal acetyl, N-terminal cap described herein, and / or C-terminal carboxamide).

[0026] In some embodiments, provided herein are compositions comprising a peptide having 4 or fewer (e.g., 4, 3, 2, 1, 0) substitutions relative to SEQ ID NO: 90; wherein the peptide comprises at least one of: Aib, D-Nle, D-Alb, or beta- Ala at AA3; Agp, Nar, Cit, or Alb at AA4; D-Fpy at AA5; or Wpb at AA7. In some embodiments, the peptide comprises 100% sequence similarity to SEQ ID NO: 90. In some embodiments, the peptide comprises SEQ ID NO: 90.

[0027] In some embodiments, provided herein are compositions comprising a peptide having 4 or fewer (e.g., 4, 3, 2, 1, 0) substitutions relative to SEQ ID NO: 91; wherein the peptide comprises at least one of: Aib, D-Nle, D-Alb, or beta- Ala at AA3; Agp, Cit, or Nar, at AA4; or Wpb at AA7. In some embodiments, the peptide comprises 100% sequence similarity to SEQ ID NO: 91. In some embodiments, the peptide comprises SEQ ID NO: 91.

[0028] In some embodiments, AA7 is Wpb.

[0029] In some embodiments, the peptide has 4 or fewer (e.g., 4, 3, 2, 1, 0) substitutions relative to one of SEQ ID NOS: 23-41, 72-80, 87-89, and / or 93-94. In some embodiments, the peptide has 100% sequence similarity with one of SEQ ID NOS: 23-41, 72-80, 87-89, and / or 93-94. In some embodiments, the peptide comprises the sequence of one of SEQ ID NOS: 23-41, 72-80, 87-89, and / or 93-94. In some embodiments, the peptide further comprises an X and / or Y cap as defined herein (e.g., N-terminal acetyl and / or C-terminal carboxamide).

[0030] In some embodiments, the peptide is a melanocortin 4 receptor (MC4R) agonist. In some embodiments, the peptide is orally bioavailable. In some embodiments, the peptide exhibits enhanced potency (e.g., greater MC4R activity or decrease in 24 hour food intake upon administration) compared to setmelanotide. In some embodiments administration of the peptide to a subject does not cause adverse cardiovascular effects. In some embodiments, a peptide exhibits two or more of the above effects.

[0031] In some embodiments, provided herein are pharmaceutical formulations comprising a composition comprising a peptide described herein and a pharmaceutically acceptable carrier. In some embodiments, the composition (or the peptide) is present at a subtherapeutic dose. In some embodiments, the composition (or the peptide) is present at a substandard dose.

[0032] In some embodiments, the pharmaceutical composition farther comprises an antiobesity agent. In some embodiments, the anti-obesity agent is selected from a GLP-1 agonist, an anorectic, a pro-satiety agent, a lipase inhibitor, and combinations thereof. In someAtorney Docket No. UM-43572.601

[0033] Client Ref. No. UM 2025-153 embodiments, the anti-obesity agent is a GLP-1 agonist. In some embodiments, the GLP-1 agonist is selected from exendin-4, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, semaglutide, taspoglutide, tirzepatide, retatrutide, CNTO736, CNT03649, HM11260C (LAPS-Exendin), NN9926 (OG9S7GT), TT401, ZYOG1, maritide, orforglipron, NN-9932, and HRS-953. In some embodiments, the anti-obesity agent is an anorectic. In some embodiments, the anorectic is selected from amphetamine, dexamphetamine, amfepramone, clobenzorex, mazindol, phentermine, phentermine and topiramate, and lorcaserin. In some embodiments, the anti-obesity agent is a pro-satiety agent. In some embodiments, the pro-satiety agent is selected from neurotrophic factor, amylin, calcitonin, cholecystokinin (CCK), leptin, metreleptin oxyntomodulin, pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), pramlitide, cagrilintide, eloralintide, petrelintide, AZD6234, GUBO 14295, and KBP336. In some embodiments, the anti-obesity agent is a lipase inhibitor. In some embodiments, the lipase inhibitor is selected from caulerpenyne, cetilistat, ebelactone A and B, esterastin, lipstatin, orlistat, percyquinin, panclicin A-E, valilactone and vibralactone. In some embodiments, the anti-obesity agent is a combination of two or more agents selected from phentermine / topiramate, naltrexone / bupropion, cagrilintide / semaglutide, amycretin, retatrutide, survodutide, pemvidutide, and mazdutide. In some embodiments, the anti-obesity agent is present at a sub-therapeutic dose. In some embodiments, the anti-obesity agent is present at a sub-standard dose. In some embodiments, the peptide is present at a sub-therapeutic dose. In some embodiments, the peptide is present at a sub-standard dose.

[0034] In some embodiments, provided herein are methods of treating a subject for a disease, condition, or disorder comprising administering a composition (e.g., comprising an MC4R agonist peptide herein) to a subject. In some embodiments, the subject suffers from positive energy balance as the cause or result of the disease, condition, or disorder. In some embodiments, the disease, condition, or disorder is characterized by overeating. In some embodiments, the disease, condition, or disorder characterized by one or more emotional / mental symptoms. In some embodiments, the disease, condition, or disorder is caused by or is the result of obesity. In some embodiments, the subject suffers from diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, and / or arthritis. In some embodiments, subject suffers from genetic obesity. In some embodiments, the subject suffers from congenital leptin deficiency, leptin receptor deficiency, MC4R deficiency, POMC deficiency, PCSK1 deficiency, LEPR deficiency, Chung-Jansen syndrome, Ulnar-Mammary syndrome, PLXNA deficiency, SRC1 deficiency, heterozygous POMC / PSK1 deficiency,Atorney Docket No. UM-43572.601

[0035] Client Ref. No. UM 2025-153 hypothalamic obesity, heterozygous LEPR deficiency, MC4R haploid insufficiency, Prader-Willi syndrome, Bardet-Biedl syndrome, Alstrom syndrome, Cohen syndrome, Smith-Magenis syndrome, 16pl 1.2 deletion syndrome, ASIP obesity syndrome, Kleine-Levin, binge eating disorders, Down syndrome, Albright’s hereditary osteodystrophy with pseudohypoparathy, loss of frmction ADCY3, SIM1 deficiency, lp36 deletion distal syndrome, or WAGR syndrome.

[0036] In some embodiments, methods comprise co-administering a peptide described herein with nutritional therapy, psychotherapy, or other pharmaceutical agents. In some embodiments, methods comprise co-administering a peptide described herein with an antiobesity agent. In some embodiments, the anti-obesity agent is selected from a GLP-1 agonist, an anorectic, a pro-satiety agent, and a lipase inhibitor. In some embodiments, the antiobesity agent is a GLP-1 agonist. In some embodiments, the GLP-1 agonist is selected from exendin-4, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, semaglutide, taspoglutide, tirzepatide, retatrutide, CNTO736, CNT03649, HM11260C (LAPS-Exendin), NN9926 (OG9S7GT), TT401, ZYOG1, maritide, orforglipron, NN-9932, andHRS-953. In some embodiments, the anti-obesity agent is an anorectic. In some embodiments, the anorectic is selected from amphetamine, dexamphetamine, amfepramone, clobenzorex, mazindol, phentermine, phentermine and topiramate, and lorcaserin. In some embodiments, the anti-obesity agent is a pro-satiety agent. In some embodiments, the pro-satiety agent is selected from neurotrophic factor, amylin, calcitonin, cholecystokinin (CCK), leptin, metreleptin oxyntomodulin, pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), pramlitide, cagrilintide, eloralintide, petrelintide, AZD6234, GUB014295, and KBP336. In some embodiments, the anti-obesity agent is a lipase inhibitor. In some embodiments, the lipase inhibitor is selected from caulerpenyne, cetilistat, ebelactone A and B, esterastin, lipstatin, orlistat, percyquinin, panclicin A-E, valilactone and vibralactone. In some embodiments, the anti-obesity agent is a combination of two or more agents, such as phentermine / topiramate, naltrexone / bupropion, cagrilintide / semaglutide, amycretin, retatrutide, survodutide, pemvidutide, and mazdutide. In some embodiments, the anti-obesity agent is administered at a sub-therapeutic dose. In some embodiments, the anti-obesity agent is administered at a sub-standard dose. In some embodiments, the peptide (e.g., MC4R agonist peptide) is administered at a sub-therapeutic dose. In some embodiments, the peptide (e.g., MC4R agonist peptide) is administered at a sub-standard dose.

[0037] In some embodiments, the administration is repeated on a recurring basis for a period of at least 1 week. In some embodiments, the administration is repeated on a daily basis. InAtorney Docket No. UM-43572.601

[0038] Client Ref. No. UM 2025-153 some embodiments, the administration is repeated on a recurring basis for a period of at least 1 month. In some embodiments, the administration is repeated on a recurring basis for a period of at least 1 year.

[0039] In some embodiments, a method results in a reduction of mass to the subject (e.g., >5% reduction in body mass, >10% reduction, >15%, >20%, >25%, >30%, or more). In some embodiments, greater than 50% of the reduction in mass is loss of fat (e.g., >50%, >60%, >70%, >80%, >90%, or more). In some embodiments, at least 25% (e.g., 25%, 30%, 40%, 50%, 60%, 70%, 80%, or more) of the reduction of mass is maintained at least 3 months (e.g., 3 months, 4 months, 5 months, 6 months, 9 months, 1 year, or more) after administering the composition to the subject (or after cessation of administering of the composition).

[0040] In some embodiments, provided herein is the use of a composition comprising an MC4R agonist peptide herein in the treatment or prevention of a condition, disease, or disorder.

[0041] In some embodiments, provided herein is the use of a composition comprising an MC4R agonist peptide as a medicament.

[0042] In some embodiments, provided herein is the use of a composition comprising an MC4R agonist peptide in the manufacture of a medicament.

[0043] In some embodiments, a peptide here comprises one or more non-proteinogenic amino acids or amino acid analogs. In some embodiments, a peptide herein is cyclic molecule.

[0044] In some embodiments, provided herein are methods of treating a subject for a disease, condition, or disorder comprising administering a composition comprising a peptide described herein to the subject. In some embodiments, the subject suffers from positive energy balance as the cause or result of the disease, condition, or disorder. In some embodiments, the composition (peptide) is administered to treat (or prevent) positive energy balance. In some embodiments, the subject suffers from a disease, condition, or disorder characterized by overeating. In some embodiments, the composition (peptide) is administered to treat (or prevent) overeating. In some embodiments, the subject suffers from a disease, condition, or disorder characterized by one or more emotional / mental symptoms. In some embodiments, the composition (peptide) is administered to treat (or prevent) one or more emotional / mental symptoms. In some embodiments, the disease, condition, or disorder is caused by or is the result of obesity. In some embodiments, the composition (peptide) is administered to treat (or prevent) obesity. In some embodiments, the subject suffers from (orAtorney Docket No. UM-43572.601

[0045] Client Ref. No. UM 2025-153 is at increased risk of) diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, and / or arthritis. In some embodiments, the composition (peptide) is administered to treat (or prevent) diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, and / or arthritis. In some embodiments, the composition is co-administered with nutritional therapy, psychotherapy, or other pharmaceutical agents. In some embodiments, administration is repeated on a recurring basis for a period of at least 1 week. In some embodiments, administration is repeated on a daily basis. In some embodiments, administration is repeated on a recurring basis for a period of at least 1 month. In some embodiments, administration is repeated on a recurring basis for a period of at least 1 year.

[0046] In some embodiments, provided herein is the use of a composition comprising a peptide described herein in the treatment or prevention of a condition, disease, or disorder. In some embodiments, provided herein is the use of composition comprising a peptide described herein as a medicament. In some embodiments, provided herein are compositions comprising a peptide described herein for use in the manufacture of a medicament.

[0047] In some embodiments, provided herein are compositions (e.g., pharmaceutical compositions) comprising a melanocortin 4 receptor (MC4R) agonist peptide. In some embodiments, a peptide of a pharmaceutical composition herein is selective for MC4R over other melanocortin receptors.

[0048] In some embodiments, the peptides herein find use in the treatment and / or prevention of various conditions, such as those related to obesity and / or overeating. In some embodiments, the MC4R agonist peptides herein find use in the treatment or prevention of obesity, diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc. In some embodiments, the subject suffers from obesity. In some embodiments, the subject suffers from dietary obesity. In some embodiments, the subject suffers from an obesity syndrome due to melanocortin-4 receptor haploinsufficiency. In some embodiments, the subject suffers from an obesity syndrome due to other genetic conditions, including those responsible for hypothalamic obesity, such as craniopharyngioma. In some embodiments, the subject is at risk of overeating or becoming obese. In some embodiments, the subject has recovered from obesity or an overeating disorder and is at risk of relapsing.

[0049] In some embodiments, provided herein are methods of treating an eating disorder comprising administering a composition (e.g., pharmaceutical compositions) comprising a peptide herein to a subject suffering from the eating disorder. In some embodiments, the eating disorder is characterized by overeating. In some embodiments, the eating disorder is characterized by one or more emotional / mental symptoms. In some embodiments, theAtorney Docket No. UM-43572.601

[0050] Client Ref. No. UM 2025-153 composition is co-administered with nutritional therapy, psychotherapy, weight-management routines, a weight-loss device, bariatric surgery, diet, other weight-loss therapeutics, etc.

[0051] In some embodiments, methods are provided in which administration of a composition (e.g., pharmaceutical compositions) comprising a peptide herein is repeated on a recurring basis for a period of at least 1 week. In some embodiments, the administration is repeated on a daily basis. In some embodiments, the administration is repeated on a recurring basis for a period of at least 1 month. In some embodiments, the administration is repeated on a recurring basis for a period of at least 1 year.

[0052] In some embodiments, provided herein is the use of a composition (e.g., pharmaceutical compositions) comprising a peptide herein in the treatment or prevention of an eating disorder. In some embodiments, provided herein is the use of a composition (e.g., pharmaceutical compositions) comprising a peptide herein as a medicament. In some embodiments, provided herein is the use of a composition (e.g., pharmaceutical compositions) comprising a peptide herein the manufacture of a medicament.

[0053] BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1. Exemplary MC4R agonist, CTX-1718, with high potency and receptorsubtype specificity. Live-cell concentration-response curves for cAMP production in clonal HEK293 cell lines expressing the human melanocortin receptors indicated. The GloSensor split-firefly luciferase reporter was used for real-time detection of receptor-induced intracellular cAMP concentrations, using the Hamamatsu FDSS plate reader. EMAX, Maximum effect relative to the native ligand a-MSH. Data points represent the mean from a representative experiment with 6 to 12 replicates. Box indicates EC50 values determined using GraphPad Prism.

[0055] Figure 2. Effect of chronic administration of courage compounds on weight loss and food intake. Fifteen-week old male C57BL / 6J mice were singly housed and placed on high-fat diets (60% kcal fat, Research Diets D12492) for 37 days. Mice were then exposed for 5 days (days 33-37), including handling and daily subcutaneous saline injections. On day 38, mice (N=7 / group) were then treated, immediately before lights out, with subcutaneous injections of vehicle (saline), semaglutide (10 nmol / kg), and / or melanocortin compound (Img / kg) as indicated in panels A-J below. Beginning on day 39, food intake and body weight were then measured at lights off, prior to injections, for 14 days in a row. Data show percent reduction in body weight (A-E), and cumulative food intake in grams (F-J).Atorney Docket No. UM-43572.601

[0056] Client Ref. No. UM 2025-153 Figure 3A-G. Molecular structures of exemplary peptides. (A) CTX-1687 (SEQ ID NO: 23) - comprises only a cyclic portion; cyclized between N-terminal S-acetyl and AA8. . (B) CTX-1691 (SEQ ID NO: 27) - comprises only a cyclic portion and a non-cyclic C-terminal tail; cyclized between N-terminal S-acetyl and AA8. (C) CTX-1661 (SEQ ID NO: 5) - comprises a cyclic portion and a non-cyclic N-terminal amino acid; cyclized between AA2 and AA8 by an -S-S- group. (D) CTX-1678 (SEQ ID NO: 18) - comprises a cyclic portion and non-cyclic N- and C-terminal amino acids; cyclized between AA2 and AA8 by an -S-S-group. (E) CTX-1682 (SEQ ID NO: 21) - comprises a cyclic portion and non-cyclic N-terminal amino acids; cyclized between AA2 and AA8 by an -S-S- group. (F) CTX-1761 (SEQ ID NO: 81) - comprises a cyclic portion, a non-cyclic N-terminal amino acid, and a phenyl-(CH2)3-C[=O] cap; cyclized between AA2 and AA8 by an -S-S- group. (G) CTX-1764 (SEQ ID NO: 83) - comprises a cyclic portion and a non-cyclic N-terminal amino acid; cyclized between AA2 and AA8 by a -S-CH2-S- group.

[0057] Figure 4. Daily subcutaneous treatment with CTX-1649 (Img / kg) and tirzepatide (2nmol / kg) or cagrilintide (lOnmol / kg).

[0058] Figure 5. Co-administration of CTX peptides, but not Setmelanotide, blunts the rate of weight regain following weight loss induced by the combination of a CTX compound plus a GLP1 drug.

[0059] Figure 6. Combination therapy with CTX-melanocortin peptides preferentially increases fat mass loss. At the conclusion of drug treatment period (left panel), body composition was determined by NMR, using a Broker minispec. Combination treatment with tirzepatide and CTX-1649 doubled fat mass loss (middle panel) compared to tirzepatide alone, with no increase in loss of lean mass (right panel). N=5.

[0060] Figure 7. CTX Peptides show significant improvement in the treatment of MC4R haploinsufficiency, relative to setmelanotide. (top panel). A subset of peptides were tested for their ability to inhibit food intake in a 24hr feeding assay in the MC4R+ / - mouse. Several peptides show greatly enhanced in vivo potency. Many peptides also exhibited significant improvement in MC4R selectivity in this assay (blue bars) relative to setmelanotide (red bar). Increased ratio of MC4R potency to MC1R potency is shown in green bars (log scale), (botom panel) One such compound identified in this manner, CTX-1715, was then tested in a 14 day weight loss assay. Using obese male MC4R+ / - mice, animals were treated daily with setmelanotide or CTX-1715 (each at Img / kg). Setmelanotide produced -2.5% weight loss, while CTX-15 yielded 12.5-15% weight loss. N=7 / group. Fat mass loss, determined by NMR, is shown on the right.Atorney Docket No. UM-43572.601

[0061] Client Ref. No. UM 2025-153 Figure 8. Cardiovascular safety of CTX peptides compared to setmelanotide. Five rhesus macaques were instrumented for measurement of blood pressure and heart rate.

[0062] Setmelanotide, CTX-1715, and CTX- 1649 were each administered 1 mg / kg as a subcutaneous injection, daily over three days. Recordings were continuous, and AUC values were calculated for systolic blood pressure and heart rate for each day. Results of each treatment were compared to control injection of saline.

[0063] Figure 9. CTX- 1649 and CTX-1715 can be detected in plasma after oral gavage in the mouse. Female CD1 mice were gavaged with CTX-1649 or CTX-1715 at 20mg / kg, and the permeabilizers indicated (n-6 per condition). Permeabilizing agents SNAC (200mg / kg) or Caprate (390mg / kg) were included. Mice were fasted overnight with access to water, and gavaged in the morning with volumes of ~300ml. Retro-orbital bleeds were taken at the times indicated, plasma prepared and analyzed by mass spectrometry. Concentrations were determined by comparing experimental samples with standard curves, created with known peptide dilutions in control plasma.

[0064] DEFINITIONS

[0065] Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments described herein, some preferred methods, compositions, devices, and materials are described herein. However, before the present materials and methods are described, it is to be understood that this invention is not limited to the particular molecules, compositions, methodologies or protocols herein described, as these may vary in accordance with routine experimentation and optimization. It is also to be understood that the terminology used in the description is for the purpose of describing the particular versions or embodiments only, and is not intended to limit the scope of the embodiments described herein.

[0066] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. However, in case of conflict, the present specification, including definitions, will control. Accordingly, in the context of the embodiments described herein, the following definitions apply.

[0067] As used herein and in the appended claims, the singular forms “a”, “an” and “the” include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to “an agonist peptide” is a reference to one or more agonist peptides and equivalents thereof known to those skilled in the art, and so forth.Atorney Docket No. UM-43572.601

[0068] Client Ref. No. UM 2025-153 As used herein, the term “comprise” and linguistic variations thereof denote the presence of recited feature(s), element(s), method step(s), etc. without the exclusion of the presence of additional feature(s), element(s), method step(s), etc. Conversely, the term “consisting of’ and linguistic variations thereof, denotes the presence of recited feature(s), element(s), method step(s), etc. and excludes any unrecited feature(s), element(s), method step(s), etc., except for ordinarily-associated impurities. The phrase “consisting essentially of’ denotes the recited feature(s), element(s), method step(s), etc. and any additional feature(s), element(s), method step(s), etc. that do not materially affect the basic nature of the composition, system, or method. Many embodiments herein are described using open “comprising” language. Such embodiments encompass multiple closed “consisting of’ and / or “consisting essentially of’ embodiments, which may alternatively be claimed or described using such language.

[0069] As used herein, the term “MC4R agonist” refers to an agent (e.g., peptide, etc.) that binds to MC4R and promotes MC4R to produce its biological activity to at least the same degree as a natural ligand for MC4R (e.g., a-melanocyte stimulating hormone (a-MSH) or adrenocorticotropic hormone). In some embodiments, an MC4R agonist binds to MC4R in the same location as a natural MC4R ligand.

[0070] As used herein, the term “CTX peptides” refers to the peptides described and within the scope herein.

[0071] As used herein, the term “subject” broadly refers to any animal, including but not limited to, human and non-human animals (e.g., dogs, cats, cows, horses, sheep, poultry, fish, crustaceans, etc.). As used herein, the term “patient” typically refers to a subject that is being treated for a disease or condition.

[0072] As used herein, the term “obesity” refers to a medical condition with excess body fat accumulation and people are generally defined to be obese when their body mass index (BMI; a value of body mass (kg) over body height squared (m)) is 30 or higher. Obesity is most commonly caused by energy imbalance due to excessive food intake compared to energy consumption over a long period of time (“positive energy balance”). Obesity, being a metabolic disease that affects the entire body, increases the possibility of developing of diabetes and hyperlipidemia, increases the risk of the incidence of sexual dysfiinction, arthritis, and cardiovascular disease, and is associated with cancer development in some cases.

[0073] As used herein, the term “subject at risk for a disease,” for example, “a subject at risk for diabetes” or “a subject at risk for hypertension” refers to a subject with one or more riskAtorney Docket No. UM-43572.601

[0074] Client Ref. No. UM 2025-153 factors (e.g., obesity, overeating, etc.) for developing the disease. Depending upon the specific disease, risk factors may include, but are not limited to, gender, age, genetic predisposition, environmental exposures, and previous incidents of diseases, lifestyle, etc.

[0075] As used herein, the term “effective amount” refers to the amount of a composition sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages and is not intended to be limited to a particular formulation or administration route.

[0076] As used herein, the terms “administration” and “administering” refer to the act of giving a drug, prodrug, or other agent, or therapeutic treatment to a subject or in vivo, in vitro, or ex vivo cells, tissues, and organs. Exemplary routes of administration to the human body can be through space under the arachnoid membrane of the brain or spinal cord (intrathecal), the eyes (ophthalmic), mouth (oral), skin (topical or transdermal), nose (nasal), lungs (inhalant), oral mucosa (buccal), ear, rectal, vaginal, by injection (e.g., intravenously, subcutaneously, intratumorally, intraperitoneally, etc.) and the like.

[0077] As used herein, the terms “co-administration” and “co-administering” refer to the administration of at least two agent(s) (e.g., an MC4R agonist and one or more additional therapeutics) or therapies to a subject. In some embodiments, the co-administration of two or more agents or therapies is concurrent (e.g., in a single formulation / composition or in separate formulations / compositions). In other embodiments, a first agent / therapy is administered prior to a second agent / therapy. Those of skill in the art understand that the formulations and / or routes of administration of the various agents or therapies used may vary. The appropriate dosage for co-administration can be readily determined by one skilled in the art. In some embodiments, when agents or therapies are co-administered, the respective agents or therapies are administered at lower dosages than appropriate for their administration alone. Thus, co-administration is especially desirable in embodiments where the co-administration of the agents or therapies lowers the requisite dosage of a potentially harmful (e.g., toxic) agent(s), and / or when co-administration of two or more agents results in sensitization of a subject to beneficial effects of one of the agents via co-administration of the other agent.

[0078] As used herein, the term “therapeutically effective dose” refers to the amount of a pharmaceutical agent sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages and is not intended to be limited to a particular formulation or administration route.Atorney Docket No. UM-43572.601

[0079] Client Ref. No. UM 2025-153 As used herein, the term “standard dose” refers to the amount of a pharmaceutical agent that is provided to a subject for the treatment / prevention of a particular condition under standard medical practice and / or regulatory guidelines. The standard dose may be defined by a range of doses and may be somewhat variable depending on patient-specific and / or condition-specific factors. For example, the standard dose may be higher for a subject with an extreme condition or with a higher body weight.

[0080] As used herein, the term “sub-therapeutic dose,” refers to a dose of an agent which, when used in a standard protocol, would elicit litle or no positive effect in the intended treatment, for example, a dose having a negligible or statistically insignificant anorectic effect.

[0081] As used herein, the term “sub-standard dose,” refers to a dose of an agent which is less than the standard, accepted, or recommended dose for a given subject for a given condition.

[0082] As used herein, the term “pharmaceutical composition” refers to the combination of an active agent with a carrier, inert or active, making the composition especially suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo.

[0083] The terms “pharmaceutically acceptable” or “pharmacologically acceptable,” as used herein, refer to compositions that do not substantially produce adverse reactions, e.g., toxic, allergic, or immunological reactions, when administered to a subject.

[0084] As used herein, the term “pharmaceutically acceptable carrier” refers to any of the standard pharmaceutical carriers including, but not limited to, phosphate buffered saline solution, water, emulsions (e.g., such as an oil / water or water / oil emulsions), and various types of weting agents, any and all solvents, dispersion media, coatings, sodium lauryl sulfate, isotonic and absorption delaying agents, disintegrants (e.g., potato starch or sodium starch glycolate), and the like. The compositions can also include stabilizers and preservatives. For examples of carriers, stabilizers, and adjuvants, see, e.g., Martin, Remington's Pharmaceutical Sciences, 15th Ed., MackPubl. Co., Easton, Pa. (1975), incorporated herein by reference in its entirety.

[0085] As used herein, the term “pharmaceutically acceptable salt” refers to any pharmaceutically acceptable salt (e.g., acid or base) of a compound of the present invention which, upon administration to a subject, is capable of providing a compound of this invention or an active metabolite or residue thereof. As is known to those of skill in the art, “salts” of the compounds of the present invention may be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric,Atorney Docket No. UM-43572.601

[0086] Client Ref. No. UM 2025-153 nitric, perchloric, fiimaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic, formic, benzoic, malonic, naphthalene-2 -sulfonic, benzenesulfonic acid, and the like. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.

[0087] As used herein, the term “instructions for administering said compound to a subject,” and grammatical equivalents thereof, includes instructions for using the compositions contained in a kit for the treatment of conditions (e.g., providing dosing, route of administration, decision trees for treating physicians for correlating patient-specific characteristics with therapeutic courses of action).

[0088] The term “amino acid” refers to natural amino acids, unnatural amino acids, and amino acid analogs, all in their D and L stereoisomers, unless otherwise indicated, if their structures allow such stereoisomeric forms. Embodiments herein refer to various amino acid abbreviations (single-leter or three-leter abbreviations) that will be understood by those in the field. Any amino acid abbreviations not defined herein refer to their field-accepted meaning. For example, “NMe” preceding an amino acid name refers to an “N-methyl” group on the amino acid, “Nle” is “norleucine,” “Abu” is “a- Aminobutyric acid,” “Aib” is “2-Aminoisobutyric acid,” “Tic” is “l,2,3,4-tetrahydroisoquinoline-3-carboxylic acid,” “Hph” is “homophenylalanine,” “Bip” is “(4-biphenyl)-L-alanine,” “D-Phe(4tBu)” is “D-4-tert-butyl-phenylalanine,” “bAla” is “beta alanine,” “hCys” is “homocysteine, ”“Hle” is “homoleucine,” “Cit” is “citrulline,” “Om” is “ornithine,” “Pal(2’)” is “3-(2-pyridyl)-alanine,” “Pal(3’)” is “3-(3-pyridyl)-alanine,”, “Pal(4’)” is “3-(4-pyridyl)-alanine,” “Dab” is “2,4-diamino-butanoic acid,” “4-guanidyl-Dab” is “2-amino-4-guanidino-butanoic acid,” “Dap” is “2-amino-proprionic acid,” “4-carbamoyl-Dab” is “2-amino-4-carbamoylamino-butanoic acid,” “3-carbamoyl-Dap” is “2-amino-3-carbamoylamino-proprionic acid,” “Agp” is “2-amino-3-guanidino-proprionic acid,” “Nar” is “norarginine” or “2-amino-4-guanidino-butanoic acid” (also abbreviated as “Agb”), “Alb” is “albizziin” or “2-amino-3-carbamoylamino-proprionic acid,” “D-Phe(4-Br)” is “D-4-bromo-phenylalanine,” “D-Phe(4-I)” is “D-4-iodo-phenylalanine,” “D-Phe(4-F)” is “D-4-fluoro-phenylalanine,” “Phe(4-Br)” is “4-bromo-phenylalanine,” “Phe(4-I)” is “4-iodo-phenylalanine,” “Phe(4-F)” is “4-fluoro-phenylalanine,” “Tyr(4-OMe)” is “4-O-methyl- tyrosine,” “Dip” is “diphenylalanine,” “aMe-D-Phe” is “alpha-methyl-D-phenylalanine” or “2-methyl-D-phenylalanine,” “D-Phe(4-NH-Atorney Docket No. UM-43572.601

[0089] Client Ref. No. UM 2025-153 Ac)” is “D-phenylalanine(4-acetylamino),” “Phe(4tBu)” is “4-tert-butyl-phenylalanine,” “D-Fpy” is “D-phenylalanine(4-cyano),” “D-Fpt” is “D-phenylalanine(4-trifluoromethyl),” “D-Fpm” is “D-phenylalanine(4-methyl),” “Pen” is “Penicillamine,” “Cpa” is “cyclopentylalanine,” “Phg” is “phenylglycine” or “2-phenyl-glycine),” “Nva” is “norvaline,” “Wpb” is “5 -bromo-tryptophan, ” and the single-leter or three-leter abbreviations for the common proteinogenic amino acids are provided below.

[0090] The term “proteinogenic amino acids” refers to the 20 amino acids coded for in the human genetic code, and includes alanine (Ala or A), arginine (Arg or R), asparagine (Asn or N), aspartic acid (Asp or D), cysteine (Cys or C), glutamine (Gin or Q), glutamic acid (Glu or E), glycine (Gly or G), histidine (His or H), isoleucine (He or I), leucine (Leu or L), Lysine (Lys or K), methionine (Met or M), phenylalanine (Phe or F), proline (Pro or P), serine (Ser or S), threonine (Thr or T), tryptophan (Trp or W), tyrosine (Tyr or Y) and valine (Vai or V). Selenocysteine and pyrrolysine may also be considered proteinogenic amino acids

[0091] The term “non-proteinogenic amino acid” refers to an amino acid that is not naturallyencoded or found in the genetic code of any organism and is not incorporated biosynthetically into proteins during translation. Non-proteinogenic amino acids may be “unnatural amino acids” (amino acids that do not occur in nature) or “naturally-occurring non-proteinogenic amino acids” (e.g., norvaline, ornithine, homocysteine, etc.). Examples of non-proteinogenic amino acids include, but are not limited to, azetidinecarboxylic acid, 2-aminoadipic acid, 3 -aminoadipic acid, beta-alanine, naphthylalanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisbutyric acid, 2-aminopimelic acid, tertiary-butylglycine, 2,4-diaminoisobutyric acid, desmosine, 2,2’-diaminopimelic acid, 2,3-diaminopropionic acid, N-ethylglycine, N-ethylasparagine, homoproline, hydroxy lysine, allo-hydroxy lysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylalanine , N-alkylglycine including N-methylglycine, N-methylisoleucine, N-alkylpentylglycine including N-methylpentylglycine. N-methylvaline, naphthylalanine, norvaline, norleucine (“Norleu”), octylglycine, ornithine, pentylglycine, pipecolic acid, thioproline, homolysine, and homoarginine. Non-proteinogenic also include D-amino acid forms of any of the amino acids herein, as well as non-alpha amino acid forms of any of the amino acids herein (beta-amino acids, gamma-amino acids, delta-amino acids, etc.), all of which are in the scope herein and may be included in peptides herein.

[0092] The term “amino acid analog” refers to an amino acid (e.g., natural or unnatural, proteinogenic or non-proteinogenic) where one or more of the C-terminal carboxy group, theAtorney Docket No. UM-43572.601

[0093] Client Ref. No. UM 2025-153 N-terminal amino group and side-chain bioactive group has been chemically blocked, reversibly or irreversibly, or otherwise modified to another bioactive group. For example, aspartic acid-(beta-methyl ester) is an amino acid analog of aspartic acid; N-ethylglycine is an amino acid analog of glycine; or alanine carboxamide is an amino acid analog of alanine. Other amino acid analogs include methionine sulfoxide, methionine sulfone, S-(carboxymethyl)-cysteine, S-(carboxymethyl)-cysteine sulfoxide and S-(carboxymethyl)-cysteine sulfone.

[0094] As used herein, the term “peptide” refers an oligomer to short polymer of amino acids linked together by peptide bonds. In contrast to other amino acid polymers (e.g., proteins, polypeptides, etc.), peptides are of about 30 amino acids or less in length. A peptide may comprise natural amino acids, non-natural amino acids, proteinogenic amino acids, non-proteinogenic amino acids, amino acid analogs, and / or modified amino acids. A peptide may be a subsequence of naturally occurring protein or a non-natural (artificial) sequence.

[0095] As used herein, the term “cyclic peptide” refers to a cyclic derivative of a peptide in which two amino acids that are not adjacent in the linear sequence are linked to form a loop in the peptide. For example, one or more additional groups suitable for cyclization may be added to facilitate cyclization of the peptide or peptide segment. A cyclic peptide may contain an intramolecular disulfide bond (e.g., -S— S-), an intramolecular amide bond between two residues, (e.g., — CONH— or — NHCO— ), an intramolecular S-alkyl bond (e.g., — S— (CH2)n— CONH— or -NH-CO(CH2)n-S-, wherein n is 1-6; -S-CH2-S-), etc.

[0096] Cyclization may be also carried out by triazine chemistry (e.g., as exemplified in Scham, D. et al. (2001) J. Org, Chem 66; 507; incorporated by reference in its entirety). Cyclic peptides or peptide segments are denoted with the prefix “cyclo” in front of the peptide sequence and the cyclic part of the sequence within parenthesis (e.g., “Arg-cyclo(Cys-D-Ala-His-D-Phe-Arg-Trp-Cys)” wherein the two Cys residues are linked to form a cyclic peptide segment of “Cys-D-Ala-His-D-Phe-Arg-Trp-Cys”).

[0097] As used herein, the term “artificial” refers to compositions and systems that are designed or prepared synthetically, and are not naturally occurring. For example, an artificial peptide, peptoid, or nucleic acid is one comprising a non-natural sequence (e.g., a peptide without 100% identity with a naturally-occurring protein or a fragment thereof).

[0098] As used herein, a “conservative” amino acid substitution refers to the substitution of an amino acid in a peptide or polypeptide with another amino acid having similar chemical properties, such as size or charge. For purposes of the present disclosure, each of theAtorney Docket No. UM-43572.601

[0099] Client Ref. No. UM 2025-153 following eight groups contains amino acids that are conservative substitutions for one another:

[0100] 1) Alanine (A) and Glycine (G);

[0101] 2) Aspartic acid (D) and Glutamic acid I;

[0102] 3) Asparagine (N) and Glutamine (Q);

[0103] 4) Arginine I and Lysine (K);

[0104] 5) Isoleucine (I), Leucine (L), Methionine (M), and Valine (V);

[0105] 6) Phenylalanine (F), Tyrosine (Y), and Tryptophan (W);

[0106] 7) Serine (S) and Threonine (T); and

[0107] 8) Cysteine I and Methionine (M).

[0108] Naturally occurring residues may be divided into classes based on common side chain properties, for example: polar positive (or basic) (histidine (H), lysine (K), and arginine I); polar negative (or acidic) (aspartic acid (D), glutamic acid I); polar neutral (serine (S), threonine (T), asparagine (N), glutamine (Q)); non-polar aliphatic (alanine (A), valine (V), leucine (L), isoleucine (I), methionine (M)); non-polar aromatic (phenylalanine (F), tyrosine (Y), tryptophan (W)); proline and glycine; and cysteine. As used herein, a “semiconservative” amino acid substitution refers to the substitution of an amino acid in a peptide or polypeptide with another amino acid within the same class.

[0109] In some embodiments, unless otherwise specified, a conservative or semiconservative amino acid substitution may also encompass non-naturally occurring amino acid residues that have similar chemical properties to the natural residue. These non-natural residues are typically incorporated by chemical peptide synthesis rather than by synthesis in biological systems. These include, but are not limited to, peptidomimetics and other reversed or inverted forms of amino acid moieties. Embodiments herein may, in some embodiments, be limited to natural amino acids, non-natural amino acids, and / or amino acid analogs.

[0110] Non-conservative substitutions may involve the exchange of a member of one class for a member from another class.

[0111] As used herein, the term “sequence identity” refers to the degree of which two polymer sequences (e.g., peptide, polypeptide, nucleic acid, etc.) have the same sequential composition of monomer subunits. The term “sequence similarity” refers to the degree with which two polymer sequences (e.g., peptide, polypeptide, nucleic acid, etc.) differ only by conservative and / or semi-conservative amino acid substitutions. The “percent sequence identity” (or “percent sequence similarity”) is calculated by: (1) comparing two optimallyAtorney Docket No. UM-43572.601

[0112] Client Ref. No. UM 2025-153 aligned sequences over a window of comparison (e.g., the length of the longer sequence, the length of the shorter sequence, a specified window, etc.), (2) determining the number of positions containing identical (or similar) monomers (e.g., same amino acids occurs in both sequences, similar amino acid occurs in both sequences) to yield the number of matched positions, (3) dividing the number of matched positions by the total number of positions in the comparison window (e.g., the length of the longer sequence, the length of the shorter sequence, a specified window), and (4) multiplying the result by 100 to yield the percent sequence identity or percent sequence similarity. For example, if peptides A and B are both 20 amino acids in length and have identical amino acids at all but 1 position, then peptide A and peptide B have 95% sequence identity. If the amino acids at the non-identical position shared the same biophysical characteristics (e.g., both were acidic), then peptide A and peptide B would have 100% sequence similarity. As another example, if peptide C is 20 amino acids in length and peptide D is 15 amino acids in length, and 14 out of 15 amino acids in peptide D are identical to those of a portion of peptide C, then peptides C and D have 70% sequence identity, but peptide D has 93.3% sequence identity to an optimal comparison window of peptide C. For the purpose of calculating “percent sequence identity” (or “percent sequence similarity”) herein, any gaps in aligned sequences are treated as mismatches at that position.

[0113] Any peptides described herein as having a particular percent sequence identity or similarity (e.g., at least 70%) with a reference sequence ID number, may also be expressed as having a maximum number of substitutions (or terminal deletions) with respect to that reference sequence. For example, a sequence having at least Y% sequence identity (e.g., 90%) with SEQ ID NO:Z (e.g., 20 amino acids) may have up to X substitutions (e.g., 2) relative to SEQ ID NO:Z, and may therefore also be expressed as “having X (e.g., 2) or fewer substitutions relative to SEQ ID NO:Z.”

[0114] DETAILED DESCRIPTION

[0115] Provided herein are melanocortin 4 receptor (MC4R) agonist peptides and methods of use thereof for the treatment and / or prevention of eating disorders (e.g., overeating), metabolic disorders (e.g., disorders resulting in positive energy imbalance), emotional / mental disorders, and / or dietary or syndromic obesity. In particular, provided herein are MC4R agonist peptides that exhibit enhanced selectivity for MC4R over other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). The peptides herein may exhibit enhanced inAtorney Docket No. UM-43572.601

[0116] Client Ref. No. UM 2025-153 vitro potency, in vivo efficacy, pharmacokinetic properties, and / or stability compared to other known melanocortin 4 receptor binding peptides.

[0117] Central regulation of feeding and body weight is primarily controlled by neural circuits located in the hypothalamus and hindbrain (Refs. 1-3; herein incorporated by reference in their entireties). The central melanocortin system, composed of a set of two neuronal cell types located in the hypothalamic arcuate nucleus, the agouti related peptide neurons (AgRP neurons) and the pro-opiomelanocortin neurons (POMC neurons), engages this hypothalamic and hindbrain circuitry to potently regulate feeding and body weight (Refs.

[0118] 4-6; herein incorporated by reference in their entireties). AgRP and POMC neurons project to largely overlapping brain regions to exert opposing effects on feeding and body weight. For example, AgRP neurons synthesize and release the melanocortin receptor antagonist / inverse agonist, agouti related peptide (AgRP), GABA, and neuropeptide Y to stimulate feeding and body weight (Ref. 7; herein incorporated by reference in its entirety). In contrast, POMC neurons synthesize and release the endogenous melanocortin receptor agonist, alpha melanocyte stimulating hormone (a-MSH), in addition to fast excitatory / inhibitory neurotransmiters to suppress feeding and reduce body weight (Refs. 4, 8; herein incorporated by reference in their entireties).

[0119] Hypothalamic AgRP neurons play a potent role in stimulating feeding (Refs. 6, 9, 10; herein incorporated by reference in their entireties). Ablation of AgRP neurons in adult mice leads to starvation and death, while stimulation rapidly and robustly stimulates food intake and body weight in sated animals (Refs. 11-13; herein incorporated by reference in their entireties). In addition to stimulating feeding, AgRP neuronal activation also suppresses competing need states, such as anxiety and fear, thereby promoting food seeking behavior in response to negative energy balance (Refs. 14-15; herein incorporated by reference in their entireties). Intense effort has focused on identifying pharmacological targets which suppress AgRP neural circuits as a potential therapeutic strategy for obesity.

[0120] The melanocortin 4 receptor (MC4R), a component of the leptin-melanocortin pathway, plays a part in bodyweight regulation (Clement et al. The Lancet. 2020 Dec;8(12):960-970.; incorporated by reference in its entirety). Pro-opiomelanocortin (POMC)-derived peptides act on neurons expressing the Melanocortin 4 receptor (MC4R) to reduce body weight. Setmelanotide is a highly potent MC4R agonist that leads to weight loss in diet-induced obese animals and in obese individuals with complete POMC deficiency (Collet et al. Mol Metab. 2017 Oct;6(10): 1321 -1329.; incorporated by reference in its entirety):Atorney Docket No. UM-43572.601

[0121] Client Ref. No. UM 2025-153

[0122]

[0123] (setmelanotide). In some embodiments, the peptides provided herein are variants of setmelanotide, having sequence similarity and / or an analogous cyclization motif. In some embodiments, peptides herein comprise amino substitutions, deletions, additions, etc. relative to setmelanotide.

[0124] Embodiments herein provide for the modulation (e.g., activation) of MC4R in order to achieve a desired impact on a condition of energy metabolism (e.g., obesity), eating habits (e.g., overeating, etc.), or downstream effect thereof (e.g., hypertension, heart disease, diabetes, etc.). In some embodiments, MC4R agonist peptides for activating MC4R are administered to a subject and / or co-administered with one or more additional therapeutics / therapies.

[0125] In some embodiments, provided herein are methods of treating, preventing, and / or ameliorating the symptoms of overeating, obesity, diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc. by enhancing the activity of MC4R in a subject via administration of a MC4R agonist peptide. As used herein, “treatment” or “treating” of a subject (e.g., a mammal, such as a human) refers to alleviating a disease or condition partially or entirely; preventing the disease or condition; decreasing the likelihood of occurrence or recurrence of the disease or condition; slowing the progression or development of the disease or condition; or eliminating, reducing, reversing, alleviating, ameliorating, inhibiting, or preventing the onset, progression, development, severity or recurrence of one or more symptoms associated with the disease or condition. Treatment may be performed either prophylactically or subsequent to the initiation of a pathologic event or contact with an etiologic agent. Also included are “prophylactic” treatments, which can be directed to reducing the rate of progression of the disease or condition being treated, delaying the onset of that disease or condition, or reducing the severity of its onset. “Treatment” or “prophylaxis” does not necessarily indicate complete eradication, cure, or prevention of the disease or condition, or associated symptoms thereof. In some embodiments, “treatment” orAtorney Docket No. UM-43572.601

[0126] Client Ref. No. UM 2025-153 “treating” includes a partial remission. In some embodiments, “treatment” or “treating” includes a complete remission.

[0127] In some embodiments, the subject suffers from overeating, obesity (e.g., dietary obesity, syndromic obesity (e.g., melanocortin-4 receptor haploinsufficiency), etc.), diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc. In some embodiments, the subject is at risk of developing obesity, diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc., or having a recurrence of one of the aforementioned conditions.

[0128] In some embodiments, provided herein are MC4R agonist peptides. In some embodiments, provided herein are peptides of the sequence X-AA0-AA1-AA1B-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9-AA10-AA11-Y (SEQ ID NO: 92); wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is acetyl, chloro acetyl, 4-I-phenyl-(CH2)3-C[=O] (or 4-I-phenyl with an alternative linker), or absent; wherein AAO is absent or bAla; wherein AA1 is Arg or absent; wherein AA1B is Nle, Phe, HpH, Gly, Ala, or absent; wherein AA2 is Cys, hCys, or absent; wherein AA3 is D-Ala, Glu, D-Glu, D-Gly, D-Aib, Gly, Ala, NMe-Ala, Aib, Abu, D-Abu, Pro, D-Nle, D-Hle, D-Alb, beta-Ala, or absent; wherein AA4 is Arg, D-Arg, NMe-Arg, NMe-D-Arg, Cit, D- Cit, His, D-His, Nme-His, NMe-D-His, Pro, D-Om, Om, Homo- Arg, Homo-Cit, Homo-D-Cit, Pal(2’), Pal(3’), Pal(4’), D-Pal(2’), D-Pal(3’), 4-guanidyl-Dab, 4-guanidyl-D-Dab, 3-guanidyl-Dap, 3-guanidyl-D-Dap, 5-carbamoyl-Dab, 5-carbamoyl-D-Dab, 3-carbamoyl-Dap, 3-carbamoyl-D-Dap, Agp, Nar, Alb, or D-Pal(4’); wherein AA5 is Phe, D-Phe, D-Phe(4-Br), D-Phe(4-I), D-Phe(4-F), D-Phe(4-tBu), Phe(4-Br), Phe(4-F), D-Tyr, Tyr(4-OMe), or D-Tyr(4-OMe), D-Hph, D-Bip, D-Tic, D-Dip, D-Trp, aMe-D-Phe, D-Phe(4-NH-Ac), NMe-D-Phe, Phe(4-tBu), Trp, Hph, Bip, Tic, Dip, aMe-Phe, Phe(4-NH-Ac), NMe-Phe, Tyr, Phe(4-I), D-Phe(4-I), Phe(4-tBu)], D-Phe(4-guanidyl), D-Fpy, D-Fpt, D-Fpm, or Phe(4-guanidyl); wherein AA6 is Arg, NMe-Arg, D-Arg, Cit, NMe-D-Arg, or D-Cit; wherein AA7 is Trp, D-Trp, NMe-Trp, Phe, D-Phe, D-Ala, D-Tic, NMe-D-Trp, Ala, Wpb, or Tic; wherein AA8 is Cys or L-Pen; wherein AA9 is Lys, Arg, Abu, Hie, Cpa, Phg, D-Hph, Nva, Om, or absent; wherein AA10 is Pro, Phe, D-Phe, or absent; wherein AA11 is Vai, Gly, or absent; wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent; wherein AA1 is linked to AA2 if AA1B is absent; wherein AA9 is present if AA10 is present; wherein AA9 and AA10 are present if AA11 is present; wherein the peptide comprises at least one of: bAla at AAO; Phe, HpH, Gly, Ala at AA1B; hCys at AA2; Pro, D-Nle, D-Hle,Atorney Docket No. UM-43572.601

[0129] Client Ref. No. UM 2025-153 D-Alb, beta- Ala at AA3; Agp, Nar, Thr, Cit, or Alb at AA4; D-Fpy, D-Fpt, D-Fpm at AA5; Wpb at AA7; L-Pen at AA8; or Abu, Hie, Cpa, Phg, D-Hph, Nva, or Om at AA9.

[0130] In some embodiments, peptides comprise conservative or semiconservative substitutions relative to SEQ ID NO: 92. In some embodiments, a conservative or semiconservative substitution may also comprise a non-proteinogenic amino acid or amino acid analog with similar characteristics.

[0131] In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) conservative sequence similarity with SEQ ID NO: 92. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) semi-conservative sequence similarity with SEQ ID NO: 92. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) sequence identity with SEQ ID NO: 92.

[0132] In some embodiments, provided herein are peptides having 4 or fewer (e.g., 4, 3, 2, 1) substitutions (e.g., conservative, semi-conservative, unconservative, etc.) relative to one or more of SEQ ID NOS: 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, and / or 111.

[0133] In some embodiments, provided herein are cyclic MC4R agonist peptides of the sequence X-AA0-AAl-AAlB-c[AA2-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y (SEQ ID NO: 1); wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is acetyl, 4-I-phenyl-(CH2)3-C[=O] (or 4-I-phenyl with an alternative linker), R1 as disclosed herein, or absent; wherein AA0 is absent or bAla; wherein AA1 is Arg or absent; wherein AA1B is Nle, Phe, Hph, Gly, Ala, D-Ala, or absent; wherein AA2 is Cys; wherein AA3 is D-Ala, Pro, D-Nle, D-Hle, Aib, Gly, beta- Ala, D-Alb, or absent; wherein AA4 is His, Alb, Nar, Arg, Cit, Om, Pal(4’), Pal(2’), Pal(3’), or homoCit; wherein AA5 is D-Phe, D-Fpy, D-Fpm, D-Fpt, D-Tyr(4-OMe), D-Phe(4-Br), or Phe; wherein AA6 is Arg; wherein AA7 is Trp, Wpb, or Phe; wherein AA8 is Cys; wherein AA9 is Nva, Hie, Cpa, Phg, Abu, Om, D-Hph, or absent; wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent; wherein AA1 is linked to AA2 if AA1B is absent; and wherein the peptide comprises at least one of: bAla at AA0; Phe, Hph, Gly, Ala atAtorney Docket No. UM-43572.601

[0134] Client Ref. No. UM 2025-153 AA1B; Pro, D-Nle, D-Hle, D-Alb, or beta-Ala at AA3; Agp, Nar, Cit, or Alb at AA4; D-Fpy, D-Fpt, or D-Fpm at AA5; Wpb at AA7; or Abu, Hie, Cpa, Phg, D-Hph, or Om at AA9.

[0135] In some embodiments, cyclic peptides comprise conservative or semiconservative substitutions relative to SEQ ID NO: 1. In some embodiments, a conservative or semiconservative substitution may also comprise a non-proteinogenic amino acid or amino acid analog with similar characteristics.

[0136] In some embodiments, provided herein are cyclic peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) conservative sequence similarity with SEQ ID NO: 1. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) semi-conservative sequence similarity with SEQ ID NO: 1. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) sequence identity with SEQ ID NO: 1.

[0137] In some embodiments, provided herein are cyclic MC4R agonist peptides of the sequence X-AA0-AAl-AAlB-c[AA2-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y (SEQ ID NO: 2); wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is acetyl, 4-I-phenyl-(CH2)3-C[=O] (or 4-I-phenyl with an alternative linker), R1 as disclosed herein, or absent; wherein AA0 is absent or bAla; wherein AA1 is Arg or absent; wherein AA1B is Nle, Phe, Hph, Gly, Ala, D-Ala, or absent; wherein AA2 is Cys; wherein AA3 is D-Ala, Pro, D-Nle, D-Hle, Aib, or absent; wherein AA4 is His, Alb, Nar, Arg, Cit, Om, Pal(4’); wherein AA5 is D-Phe, D-Fpy, D-Fpm, D-Fpt; wherein AA6 is Arg; wherein AA7 is Trp or Wpb; wherein AA8 is Cys; wherein AA9 is Hie, Cpa, Phg, Abu, Om, D-Hph, or absent; wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent; wherein AA1 is linked to AA2 if AA1B is absent; wherein the peptide comprises at least one of: bAla at AA0; Phe, Hph, Gly, Ala at AA1B; Pro, D-Nle, D-Hle, D-Alb, or beta-Ala at AA3; Agp, Nar, Cit, or Alb at AA4; D-Fpy, D-Fpt, or D-Fpm at AA5; Wpb at AA7; or Abu, Hie, Cpa, Phg, D-Hph, or Om at AA9.

[0138] In some embodiments, cyclic peptides comprise conservative or semiconservative substitutions relative to SEQ ID NO: 2. In some embodiments, a conservative or semiconservative substitution may also comprise a non-proteinogenic amino acid or amino acid analog with similar characteristics.

[0139] In some embodiments, provided herein are cyclic peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) conservative sequence similarity with SEQ ID NO: 2. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) semi-conservative sequence similarity withAtorney Docket No. UM-43572.601

[0140] Client Ref. No. UM 2025-153 SEQ ID NO: 2. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) sequence identity with SEQ ID NO: 2.

[0141] In some embodiments, provided herein are cyclic peptides having 4 or fewer (e.g., 4, 3, 2, 1) substitutions (e.g., conservative, semi-conservative, unconservative, etc.) relative to one or more of SEQ ID NOS: 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 81, 82, 83, 84, 85, 86, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, and / or 111 wherein the peptides are cyclized between AA8 and AA2.

[0142] In some embodiments, provided herein are cyclic MC4R agonist peptides of the sequence c[X-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y (SEQ ID NO: 90); wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is S-acetyl; wherein AA3 is Aib, D-Ala, Gly, D-Nle, D-Alb, or beta- Ala; wherein AA4 is Arg, Cit, His, Nar, Alb, Om, Pal(4’), homoCit, Pal(2’), Pal(3’); wherein AA5 is D-Phe, D-Fpy, D-Fpm, D-Fpt, D-Tyr(4-OMe), D-Phe(4-Br), or Phe; wherein AA6 is Arg; wherein AA7 is Trp, Wpb, D-Trp, or Phe; wherein AA8 is Cys; wherein AA9 is Nva, Hie, Cpa, Phg, Abu, Om, D-Hph, or absent; wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent; wherein the peptide comprises at least one of: D-Nle, D-Alb, or beta-Ala at AA3; Agp, Nar, Cit, or Alb at AA4; D-Fpy at AA5; or Wpb at AA7.

[0143] In some embodiments, cyclic peptides comprise conservative or semiconservative substitutions relative to SEQ ID NO: 90. In some embodiments, a conservative or semiconservative substitution may also comprise a non-proteinogenic amino acid or amino acid analog with similar characteristics.

[0144] In some embodiments, provided herein are cyclic peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) conservative sequence similarity with SEQ ID NO: 90. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) semi-conservative sequence similarity with SEQ ID NO: 90. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) sequence identity with SEQ ID NO: 90.

[0145] In some embodiments, provided herein are cyclic MC4R agonist peptides of the sequence c[X-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y (SEQ ID NO: 91); wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is S-Atorney Docket No. UM-43572.601

[0146] Client Ref. No. UM 2025-153 acetyl; wherein AA3 is Aib, D-Ala, Gly, D-Nle, D-Alb, or beta- Ala; wherein AA4 is Arg, Cit, His, or Nar; wherein AA5 is D-Phe; wherein AA6 is Arg; wherein AA7 is Trp or Wpb; wherein AA8 is Cys; wherein AA9 is Nva, Hie, Cpa, Phg, or absent; wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent; wherein the peptide comprises at least one of: D-Nle, D-Alb, or beta- Ala at AA3; Agp, Cit, or Nar, at AA4; or Wpb at AA7.

[0147] In some embodiments, cyclic peptides comprise conservative or semiconservative substitutions relative to SEQ ID NO: 91. In some embodiments, a conservative or semiconservative substitution may also comprise a non-proteinogenic amino acid or amino acid analog with similar characteristics.

[0148] In some embodiments, provided herein are cyclic peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) conservative sequence similarity with SEQ ID NO: 91. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) semi-conservative sequence similarity with SEQ ID NO: 91. In some embodiments, provided herein are peptides having at least 70% (e.g., >70%, >75%, >80%, >85%, >90%, >95%, 100%) sequence identity with SEQ ID NO: 91.

[0149] In some embodiments, provided herein are peptides having 4 or fewer (e.g., 4, 3, 2, 1) substitutions (e.g., conservative, semi-conservative, unconservative, etc.) relative to one or more of SEQ ID NOS: 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 72, 73, 74, 75, 76, 77, 78, 79, 80, 87, 88, 89, 93, and / or 94.

[0150] In some embodiments, an N-terminal cap moiety is linked to the most N-terminal amino acid of the peptide. In some embodiments, the N-terminal cap moiety is an acetyl group. In some embodiments, the N-terminal cap moiety is 4-I-phenyl-(CH2)3-C[=O] (or 4-1-phenyl with an alternative linker). In some embodiments, the N-terminal cap moiety is a chloro acetyl group. In embodiments in which the N-terminal cap is a chloro acetyl group, the N-terminal cap may react with a Cys residue within the peptide (e.g., AA8 of SEQ ID NOS: 90 and 91) to form a thio ether linkage, resulting in a thio-ether cyclized peptide (a cyclic peptide). In some embodiments, the N-terminal cap moiety is a pharmacokinetic (PK) modifying group. Such groups are described in Refs. 59-61 (incorporated by reference in their entireties) and include groups comprising a PK-modifying moiety (e.g., 4-I-phenyl, 4-1-phenyl-(CH2)3-C[=O], Cl 8 diacid, etc.), an amino acid linker moiety (e.g., Gly, yGlu, etc.), a PEG moiety (e.g., methoxy PEG (e.g., mPEG-2, mPEG-3, mPEG-4, mPEG-5, mPEG-6, orAtorney Docket No. UM-43572.601

[0151] Client Ref. No. UM 2025-153 more)), or a combination thereof. Non-limiting examples of an N-terminal PK modifying group include 4-I-phenyl-(CH2)3-C[=O]-PEG-Gly.

[0152] In some embodiments, an N-terminal cap moiety is of the general structure:

[0153]

[0154] or

[0155]

[0156] A - 0, 1 or 2 c - Q, 1 or 2

[0157] Non-limiting examples of an N-terminal cap moiety having such a general structure include Cl 8 diacid-yGlu-mPEG2 (“PKcapl” or “Rl”) and Aryl(4-I)- Gly-mPEG2 (“PKcap2” or “R2”):

[0158]

[0159] Aryl(4-I) Gly mPEG2

[0160] Other PK-modifying caps are within the scope herein.

[0161] In some embodiments, peptides herein comprise natural amino acids, unnatural amino acids, modified amino acids, non-proteinogenic amino acids, amino acid analogs, etc.

[0162] In some embodiments, all or a portion of the peptide is cyclic. For example, the exemplary peptides provided in Table 1 and Table 2, as depicted in those tables, comprise a cyclic portion.

[0163] In some embodiments of cyclic MC4R agonist peptides as exemplified in SEQ ID NOS: 1 and 2, the amino acid corresponding to AA2 is Cys, the amino acid corresponding to AA8 is Cys, AA2 is linked to AA8 through a disulfide bond (e.g., -S-S-) or a dithiol bond (e.g., -S-CH2-S-), and the peptide segment corresponding to AA2-AA8 of SEQ ID NOS: 1Atorney Docket No. UM-43572.601

[0164] Client Ref. No. UM 2025-153 and 2 is cyclic. Peptides in Tables 1 and 2 having a pair of Cys* residues comprise a -S-CH2-S- linkage; although, in certain embodiments, any of the peptides within the scope herein cyclized by a disulfide bond (e.g., -S-S-), a dithiol bond (e.g., -S-CH2-S-), or any other suitable linkage. In some embodiments, the peptide farther comprises an amino acid corresponding to AAO, AA1 or AA1B of SEQ ID NOS: 1 and 2 linked to the amino acid corresponding to AA2 of SEQ ID NOS: 1 and 2, and / or an amino acid corresponding to AA9 of SEQ ID NOS: 1 and 2 linked to the amino acid corresponding to AA8 of SEQ ID NOS: 1 and 2.

[0165] In some embodiments of cyclic MC4R agonist peptides as exemplified in SEQ ID NOS: 90 and 91, the S-acetyl (or other suitable group) corresponding to X and the Cys amino acid corresponding to AA8 are linked to form a cyclic peptide. In some embodiments, the peptide further comprises an amino acid corresponding to AA9 of SEQ ID NOS: 90 and 91 linked to the amino acid corresponding to AA8.

[0166] In some embodiments, the peptides of Table 1 and Table 2, or other peptides within the scope herein (e.g., corresponding to SEQ ID NOS: 1-111) may be provided as linear peptides. In some embodiments, peptides herein (e.g., peptides corresponding SEQ ID NOS: 1-111, peptides of Table 1, 2, etc.) are cyclic but comprise an alternative cyclization. For example, insertion or substitution of Cys or D-Cys, or other suitable residues between or for residues in the peptides allow for cyclization of a peptide segment other than AA2-AA8 (e.g., X-AA8, AA3-AA8, etc.). In some embodiments, a peptide herein may be cyclized by methods understood in the field and described herein. For example, a first amino acid within the sequence of the peptide may be substituted for a Cys or D-Cys, the amino acid corresponding to a second amino of the peptide may be substituted for a Cys, Om, or D-Cys, and the amino acid corresponding to first amino acid is linked to the second amino acid, rendering the peptide segment between those amino acids of the peptide cyclic. Any pairs of amino acids within the peptides herein can be used for cyclization. In some embodiments, any of AA1, AA1B, AA2, AA3, AA4, AA5, AA6, AA7, AA8, and AA9 may be amino acids capable of forming a cyclic section of peptide. For example, a five-amino-acid cyclic segment may be formed between AA1 and AA5, AA1B and AA6, AA2 and AA7, AA3 and AA8, AA4 and AA9. In some embodiments, a four-amino-acid cyclic segment may be formed between AA1 and AA4, AA1B and AA5, AA2 and AA6, AA3 and AA7, AA4 and AA8, AA5 and AA9. In some embodiments, a six-amino-acid cyclic segment may be formed between AA1 and AA6, AA1B and AA7, AA2 and AA8, AA3 and AA9. In some embodiments, the endpoint amino acids of the cyclic portion are Cys or D-Cys and Cys, Om,Atorney Docket No. UM-43572.601

[0167] Client Ref. No. UM 2025-153 or D-Cys. Therefore, in some embodiments, any of AA1-AA9 may be Cys, Om, or D-Cys in cyclic peptide, depending on the location of the cyclic portion. Other cyclic connections are within the scope herein.

[0168] In some embodiments, a cyclic peptide comprises a linking moiety between the linked amino acids (e.g., .. ,-S-(linking moiety)-S-...). Exemplary linking moieties include a methyl (-CH2-) group or ethyl (-(CH2)2-) group.

[0169] In some embodiments, a MC4R agonist peptide descried herein is linked at the N-and / or C-terminus to one or more additional amino acids, peptides, proteins, or other carriers. In some embodiments, provided herein is a fusion of a MC4R agonist peptide descried herein with one or more additional peptide or polypeptide sequences. In some embodiments, an additional peptide or polypeptide fused to the MC4R agonist peptide is a carrier that confers solubility, localization (within a cell, tissue, subject, etc.), stability, cell permeability, etc. In some embodiments, an additional peptide or polypeptide fiised to the MC4R agonist peptide is a therapeutic peptide or polypeptide. In some embodiments, any 1-10 additional amino acids may be fiised to the N-terminus or C-terminus of a MC4R agonist peptide descried herein. In some embodiments, a peptide or polypeptide of 10-50, 50-100, 100-200, or more amino acids is fused to the N-terminus or C-terminus of a MC4R agonist peptide descried herein.

[0170] In some embodiments, a MC4R agonist is administered to a subject (e.g., by any suitable route of administration and within any suitable pharmaceutical formulation). In some embodiments, the MC4R agonist peptide binds to MC4R in the subject. In some embodiments, the MC4R agonist peptide binds to MC3R in the subject. In some embodiments, the activity of MC4R is enhanced (MC4R is activated) by the administration of the MC4R agonist peptide.

[0171] In some embodiments, methods herein comprise administering a MC4R agonist peptide to a subject at risk of and / or suffering from overeating, obesity (e.g., dietary obesity, syndromic obesity (e.g., melanocortin-4 receptor haploinsufficiency), etc.), diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc. In some embodiments, administration of the MC4R agonist peptide results in decreased eating, decreased bodyweight, and / or other changes in observable / measurable characteristic / biomarkers for obesity, diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc.

[0172] In some embodiments, the MC4R agonist peptide is administered locally. In some embodiments, the MC4R agonist peptide is administered systemically. In some embodiments,Atorney Docket No. UM-43572.601

[0173] Client Ref. No. UM 2025-153 the MC4R agonist peptide is administered in a manner such that it reaches and / or localizes in the brain. In some embodiments, the MC4R agonist is administered in a manner such that it reaches and / or localizes in the hypothalamus. In some embodiments, the MC4R agonist peptide is administered in a manner such that it reaches and / or localizes in AgRP neurons. In some embodiments, the MC4R agonist peptide is administered in a manner such that it reaches and / or localizes in POMC neurons.

[0174] In some embodiments, a MC4R agonist peptide binds MC4R selectively over other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). In some embodiments, a MC4R agonist peptide binds MC4R with an affinity that is at least 2-fold greater (e.g., 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, lOx, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, lOOx, 200x, 500x, lOOOx, 2000x, 5000x, or more) than the binding affinity of the MC4R agonist peptide with one or more other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). In some embodiments, a MC4R agonist peptide herein binds MC4R selectively over MC3R. In some embodiments, a MC4R agonist peptide binds MC4R with an affinity that is at least 2-fold greater (e.g., 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, lOx, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, lOOx, 200x, 500x, lOOOx, 2000x, 5000x, or more) than the binding affinity of the MC4R agonist with MC3R.

[0175] In some embodiments, a MC4R agonist peptide binds MC4R selectively over other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). In some embodiments, a MC4R agonist peptide binds MC4R with an affinity that is at least 2-fold greater (e.g., 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, lOx, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, lOOx, 200x, 500x, lOOOx, 2000x, 5000x, or more) than the binding affinity of the MC4R agonist peptide with one or more other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R).

[0176] In some embodiments, a peptide herein is a MC4R agonist and activates MC4R selectively over one or more other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). In some embodiments, a peptide herein is a MC4R agonist and activates MC4R at least 2-fold greater (e.g., 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, lOx, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, lOOx, 200x, 500x, lOOOx, 2000x, 5000x, or more) than one or more other melanocortin receptors (e.g., MC1R, MC2R, MC3R, MC5R). In some embodiments, a peptide herein is a MC4R agonist and enhances the activity of MC4R selectively over MC3R. In some embodiments, a peptide herein is a MC4R agonist and activates MC4R at least 2-fold greater (e.g., 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, lOx, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, lOOx, 200x, 500x, lOOOx, 2000x, 5000x, or more) than MC3R.Atorney Docket No. UM-43572.601

[0177] Client Ref. No. UM 2025-153 In some embodiments, a MC4R agonist peptide is co-administered with an additional agent or therapy. In some embodiments, the MC4R agonist peptides described herein are anorexia-sensitizing agents. As described in Inti. Pat. App. No. PCT / US2024 / 042767 (incorporated by reference in its entirety), administration of an activator of MC4R, even at a sub-therapeutic or sub-standard dose (e.g., below a dose capable of producing a statistically significant reduction in food intake, body weight, etc.), sensitizes a subject to anti-obesity and / or anorectic agents, such that co-administration of a (1) MC4R activator with (2) an antiobesity and / or anorectic agent enhances the efficacy of the anti-obesity and / or anorectic agent, allowing for greater benefit and / or reduced dose (e.g., with reduced side effects) from the anti-obesity and / or anorectic agent. Therefore, in certain embodiments, a subtherapeutic (or substandard) dose of an MC4R agonist peptide is co-administered with a standard or therapeutic dose of an anti-obesity and / or anorectic agent to produce reduced side effects from the MC4R agonist and / or enhanced effects (e.g., greater reduction in food intake, body weight, etc.). In other embodiments, a standard or therapeutic dose an MC4R agonist peptide is co-administered with a substandard or subtherapeutic dose of an anti-obesity and / or anorectic agent to produce reduced side effects from the anti-obesity and / or anorectic agent and / or enhanced effects (e.g., greater reduction in food intake, body weight, etc.). In some embodiments, a subtherapeutic (or substandard) dose of an MC4R agonist peptide is coadministered with a substandard or subtherapeutic dose of an anti-obesity and / or anorectic agent to produce reduced side effects and / or enhanced effects (e.g., greater reduction in food intake, body weight, etc.).

[0178] In some embodiments, a MC4R agonist peptide is co-administered with an additional agent or therapy. In some embodiments, the co-administered agent is for the treatment or prevention of the same condition / disease / symptom as the MC4R agonist peptide (e.g., overeating, obesity, diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc.). In some embodiments, the co-administered agent is for the treatment or prevention of a side-effect of the MC4R agonist peptide. In some embodiments, the coadministered agent is for the treatment or prevention of a comorbidity not treated of prevented by the MC4R agonist peptide.

[0179] In some embodiments, the MC4R agonist peptide is co-administered with psychotherapy. The term "psychotherapy" refers to use of non-pharmacological therapies a clinician or therapist uses any of a variety of techniques that involve verbal and other interactions with a patient to affect a positive therapeutic outcome. Such techniques include, but are not limited to, behavior therapy, cognitive therapy, psychodynamic therapy,Atorney Docket No. UM-43572.601

[0180] Client Ref. No. UM 2025-153 psychoanalytic therapy, group therapy, family counseling, art therapy, music therapy, vocational therapy, humanistic therapy, existential therapy, transpersonal therapy, clientcentered therapy (also called person-centered therapy), Gestalt therapy, biofeedback therapy, rational emotive behavioral therapy, reality therapy, response based therapy, Sandplay therapy, status dynamics therapy, hypnosis and validation therapy. Any suitable psychotherapy techniques, including those listed above, may be co-administered with a MC3R antagonist or partial agonist peptide for the treatment / prevention of appropriate conditions / diseases (e.g., overeating, obesity, diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, arthritis, etc.).

[0181] In some embodiments, the MC4R agonist peptide is co-administered with one or more drugs for treating or treating / preventing obesity, preventing weight gain or overeating, or inducing weight loss, such as semaglutide (WEGOVY, OZEMPIC, RYBELSUS), dulaglutide (TRULICITY), phentermine (ADIPEX, IONAMIN, SUPRENZA), diethylpropion, Phentermine-Topiramate extended release (QSYMIA), Bupropion / Naltrexone (CONTRA VE), Liraglutide (SAXENDA, VICTOZA), exenatide (BYETTA, BYDUREON), lixisenatide (ADLYXIN), RESOIO, ST4-001, iBIO-610, etc.

[0182] In some embodiments, the MC4R agonist peptide is co-administered with one or more anti-obesity agents, such as a GLP-1 agonist. Suitable GLP-1 agonists for use in the compositions and methods herein include, but are not limited to, exendin-4, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, semaglutide, taspoglutide, tirzepatide, retatrutide, CNTO736, CNT03649, HM11260C (LAPS-Exendin), NN9926 (OG9S7GT), TT401, ZYOG1, Maritide, orforglipron, NN-9932, andHRS-953.

[0183] In some embodiments, the MC4R agonist peptide is co-administered with one or more anti-obesity agents, such as an anorectic. Suitable anorectics for use in the compositions and methods herein include, but are not limited to, amphetamine, dexamphetamine, amfepramone, clobenzorex, mazindol, phentermine (with or without topiramate) and lorcaserin.

[0184] In some embodiments, the MC4R agonist peptide is co-administered with one or more anti-obesity agents, such as a pro-satiety agent. Suitable pro-satiety agents for use in the compositions and methods herein include, but are not limited to, neurotrophic factor (e.g., axokine) and longer-acting analogs of amylin, calcitonin, cholecystokinin (CCK), leptin, metreleptin oxyntomodulin, pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), pramlitide, cagrilintide, eloralintide, petrelintide, AZD6234, GUB014295, KBP336, and variants thereof (e.g., PYY3-36). In some embodiments, the MC4R agonist peptide is co-Atorney Docket No. UM-43572.601

[0185] Client Ref. No. UM 2025-153 administered with an amylin analog, such as pramlitide, cagrilintide, eloralintide, petrelintide, AZD6234, GUB014295, KBP336, etc. In some embodiments, a pro-satiety agent (e.g., CCK) is co-administered with an inhibitor of MC3R expression or activity.

[0186] In some embodiments, the MC4R agonist peptide is co-administered with one or more anti-obesity agents, such as a lipase inhibitor. Suitable lipase inhibitors for use in the compositions and methods herein include, but are not limited to, caulerpenyne, cetilistat, ebelactone A and B, esterastin, lipstatin, orlistat, percyquinin, panclicin A-E, valilactone and vibralactone. In some embodiments, a lipase inhibitor (e.g., lipstatin) is co-administered with an inhibitor of MC3R expression or activity.

[0187] In some embodiments, the MC4R agonist peptide is co-administered with one or more anti-obesity agents, such as a leptin receptor agonist. Suitable leptin agonists include, but are not limited to leptin, metraleptin, and leptin receptor agonist antibodies.

[0188] In some embodiments, the MC4R agonist peptide is co-administered with one or more combination weight loss agents, such as phentermine / topiramate (Qsymia), naltrexone / bupropion, cagrilintide / semaglutide (CagriSema), amycretin, retatrutide, survodutide, pemvidutide, mazdutide, etc.

[0189] In some embodiments, a MC4R agonist is co-administered with an antidepressant agent. Suitable antidepressants for co-administration may include serotonin and noradrenaline reuptake inhibitors (e.g., duloxetine (Cymbalta), venlafaxine (Effexor), desvenlafaxine (Pristiq), etc.), selective serotonin reuptake inhibitors (e.g., italopram (Celexa), escitalopram (Lexapro), fluoxetine (Prozac, Sarafem), fluvoxamine (Luvox), paroxetine (Paxil), sertraline (Zoloft), etc.), tricyclic antidepressants (e.g., amitriptyline (Elavil), amoxapine- clomipramine (Anafranil), desipramine (Norpramin), doxepin (Sinequan), imipramine (Tofranil), nortriptyline (Pamelor), protriptyline (Vivactil), trimipramine (Surmontil), etc.), monoamine oxidase inhibitors (e.g., phenelzine (Nardil), tranylcypromine (Parnate), isocarboxazid (Marplan), selegiline (EMSAM, Eldepryl), etc.), noradrenaline and specific serotoninergic antidepressants (e.g., Mianserin (Tolvon), Mirtazapine (Remeron, Avanza, Zispin, etc.), etc.

[0190] In some embodiments, a MC4R agonist peptide is co-administered with an antianxiety agent. Suitable antianxiety medications for co-administration may include selective serotonin reuptake inhibitors, serotonin-norepinephrine reuptake inhibitors, tricyclics, benzodiazepines (e.g., alprazolam (Xanax), chlordiazepoxide (Librium), diazepam (Valium), lorazepam (Ativan) etc.), beta-blockers (e.g., atenolol (Tenormin), propranolol (Inderal), etc.), buspirone (BuSpar), monoamine oxidase inhibitors, etc.Atorney Docket No. UM-43572.601

[0191] Client Ref. No. UM 2025-153 In some embodiments, a MC4R agonist peptide is co-administered with a mood stabilizer. Suitable mood stabilizers for co-administration may include lithium, anticonvulsants (e.g., valproate, lamotrigine, carbamazepine, etc.), etc.

[0192] In some embodiments, any suitable routes and / or modes of administering the agents herein (e.g., MC4R agonist, co-administered agent, etc.) find use in embodiments herein. In some embodiments, the compositions and methods described herein act upon the central nervous system (CNS) and therefore routes and / or modes of administration that facilitate entry of the agents into the CNS are utilized. In some embodiments, the compositions and methods described herein act upon the brain of a subject and therefore routes and / or modes of administration that facilitate entry of the agents into the brain (e.g., allow agents to cross the blood-brain barrier) are utilized. In some embodiments, the compositions and methods described herein act upon the hypothalamus of a subject and therefore routes and / or modes of administration that facilitate delivery of the agents to the hypothalamus are utilized. In some embodiments, the compositions and methods described herein act upon the arcuate nucleus of the hypothalamus of a subject and therefore routes and / or modes of administration that facilitate delivery of the agents to the arcuate nucleus are utilized. In some embodiments, the compositions and methods described herein act upon the AgRP neurons of a subject and therefore routes and / or modes of administration that facilitate delivery of the agents to AgRP neurons are utilized. In some embodiments, the compositions and methods described herein act upon the POMC neurons of a subject and therefore routes and / or modes of administration that facilitate delivery of the agents to POMC neurons are utilized.

[0193] In some embodiments, routes of administration, formation of the desired agent, and the pharmaceutical composition are selected to provide efficient and effective delivery. In some embodiments, the therapeutic agents herein (e.g., MC4R agonist peptide, coadministered agent, etc.) are provided in pharmaceutical formulations for administration to a subject by a suitable route. The pharmaceutical formulations described herein can be administered to a subject by multiple administration routes, including but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular), intranasal, buccal, topical, rectal, or transdermal administration routes. Moreover, the pharmaceutical compositions described herein (e.g., comprising a MC3R antagonist or partial agonist peptide, a co-administered agent, etc.) are formulated into any suitable dosage form, including but not limited to, aqueous oral dispersions, liquids, gels, syrups, elixirs, slurries, suspensions, aerosols, fast melt formulations, effervescent formulations, lyophilized formulations, tablets, powders, pills, dragees, and capsules.Atorney Docket No. UM-43572.601

[0194] Client Ref. No. UM 2025-153 In some embodiments, the therapeutic agents herein (e.g., MC4R agonist peptide, coadministered agent, etc.) are formulated for administration to a subject. Any suitable pharmaceutical formulations may find use in embodiments herein. In some embodiments, therapeutic agents herein (e.g., MC4R agonist peptide, co-administered agent, etc.) are formulated for controlled drug release (e.g., in microparticeles (e.g., PLGA microspheres). For example, peptides can be incorporated into PLGA microspheres via techniques such as solvent evaporation, solvent diffusion, or emulsion-based methods. Such formulation may provide gradual release over extended periods (days to months), minimized need for frequent administration, protection of the peptide from enzymatic degradation, and / or targeted delivery. In some embodiments, therapeutic agents herein (e.g., MC4R agonist peptide, coadministered agent, etc.) are formulated in lipid-based carriers. In some embodiments, peptides are encapsulated in the aqueous core of the liposome (for hydrophilic peptides) or in the lipid bilayer (for hydrophobic peptides). Liposomes can be produced by methods such as reverse-phase evaporation, thin-film hydration, or extrusion. In some embodiments, therapeutic agents herein (e.g., MC4R agonist peptide, co-administered agent, etc.) are formulated in nanoparticles, such as polymeric nanoparticles, solid lipid nanoparticles (SLNs), gold nanoparticles, etc. Peptides can be encapsulated or conjugated onto nanoparticles using methods like nanoprecipitation, solvent evaporation, or surface fimctionalization techniques. The particle size can range from 10 nm to several micrometers, depending on the delivery system's design. In some embodiments, therapeutic agents herein (e.g., MC4R agonist peptide, co-administered agent, etc.) are formulated within hydrogels. Hydrogels are a versatile class of materials used for drug delivery, offering a hydrated, biocompatible environment for peptide encapsulation. Hydrogels can be made from natural polymers (e.g., alginate, chitosan, hyaluronic acid) or synthetic polymers (e.g., poly(ethylene glycol), poly(N-isopropylacrylamide)). Peptides are loaded into the hydrogel network either by physical entrapment or chemical conjugation.

[0195] One may administer the compounds and / or compositions in a local rather than systemic manner, for example, via injection of the compound directly into an organ or tissue, often in a depot preparation or sustained release formulation. Such long-acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Furthermore, one may administer the drug in a targeted drug delivery system, for example, in a liposome coated with organ-specific antibody. The liposomes will be targeted to and taken up selectively by the organ. In addition, the drug may be provided inAtorney Docket No. UM-43572.601

[0196] Client Ref. No. UM 2025-153 the form of a rapid release formulation, in the form of an extended-release formulation, or in the form of an intermediate release formulation.

[0197] Pharmaceutical preparations for oral use can be obtained by mixing one or more solid excipients with the therapeutic agent (e.g., MC4R agonist peptide, a co-administered agent, etc.) with any suitable substituents and functional groups disclosed herein, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets, pills, or capsules. Suitable excipients include, for example, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. If desired, disintegrating agents may be added, such as the cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0198] In some embodiments, agents are delivered by inhalation. For administration by inhalation, the agents described herein (e.g., MC4R agonist peptide, a co-administered agent, etc.) may be in a form as an aerosol, a mist or a powder. In some embodiments, pharmaceutical compositions described herein are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount.

[0199] Buccal formulations that include the agents described herein (e.g., MC4R agonist peptide, a co-administered agent, etc.) may be administered using a variety of formulations which include, but are not limited to, U.S. Pat. Nos. 4,229,447, 4,596,795, 4,755,386, and 5,739,136.

[0200] In some embodiments, the agents described herein (e.g., MC4R agonist peptide, a coadministered agent, etc.) are delivered transdermally. Transdermal formulations described herein may be administered using a variety of devices including but not limited to, U.S. Pat. Nos. 3,598,122, 3,598,123, 3,710,795, 3,731,683, 3,742,951, 3,814,097, 3,921,636, 3,972,995, 3,993,072, 3,993,073, 3,996,934, 4,031,894, 4,060,084, 4,069,307, 4,077,407, 4,201,211, 4,230,105, 4,292,299, 4,292,303, 5,336,168, 5,665,378, 5,837,280, 5,869,090, 6,923,983, 6,929,801 and 6,946,144; incorporated by reference in their entireties.Atorney Docket No. UM-43572.601

[0201] Client Ref. No. UM 2025-153 In some embodiments, the agents described herein (e.g., MC4R agonist peptide, a coadministered agent, etc.) are delivered by parenteral administration (e.g., intramuscular, subcutaneous, intravenous, epidural, intracerebral, intracerebroventricular, etc.).

[0202] Formulations suitable for parenteral administration may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or vehicles including water, ethanol, polyols (propyleneglycol, polyethylene-glycol, glycerol, cremophor and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Agents described herein (e.g., MC4R agonist peptide, a co-administered agent, etc.) may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank’s solution, Ringer’s solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally recognized in the field. For other parenteral injections, appropriate formulations may include aqueous or nonaqueous solutions, preferably with physiologically compatible buffers or excipients. Such excipients are generally recognized in the field.

[0203] In certain embodiments, delivery systems for pharmaceutical agents (e.g., MC4R agonist, a co-administered agent, etc.) may be employed, such as, for example, liposomes and emulsions. In certain embodiments, compositions provided herein also include an mucoadhesive polymer, selected from among, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate and dextran.

[0204] In some embodiments, an agent (e.g., MC4R agonist peptide, a co-administered agent, etc.) is administered in a therapeutically effective amount. Thus, a therapeutically effective amount is an amount that is capable of at least partially preventing or reversing a disease, disorder, or symptoms thereof. The dose required to obtain an effective amount may vary depending on the agent, formulation, disease or disorder, and individual to whom the agent is administered.

[0205] Determination of effective amounts may involve in vitro assays in which varying doses of agent are administered to cells in culture and the concentration of agent effective forAtorney Docket No. UM-43572.601

[0206] Client Ref. No. UM 2025-153 ameliorating some or all symptoms is determined in order to calculate the concentration required in vivo. Effective amounts may also be based in in vivo animal studies.

[0207] Pharmaceutical compositions may be in unit dosage forms suitable for single administration of precise dosages. In unit dosage form, the formulation is divided into unit doses containing appropriate quantities of one or more agents (e.g., MC4R agonist peptide, a co-administered agent, etc.).

[0208] Dosing and administration regimes are tailored by the clinician, or others skilled in the pharmacological arts, based upon well-known pharmacological and therapeutic considerations including, but not limited to, the desired level of therapeutic effect, and the practical level of therapeutic effect obtainable.

[0209] In some embodiments, and upon the clinician’s discretion, the administration of the compounds may be administered for an extended period of time, including throughout the duration of the patient’s life in order to treat the disorder or ameliorate or otherwise control or limit the symptoms of the patient’s disease.

[0210] In a case wherein the patient’s status does improve, upon the clinician’s discretion the administration of the agents (e.g., MC4R agonist peptide, a co-administered agent, etc.) may be given continuously; alternatively, the dose of drug being administered may be temporarily reduced or temporarily suspended for a certain length of time (i.e., a “drug holiday”). The length of the drug holiday can vary between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dose reduction during a drug holiday may be from about 10% to about 100%, including, by way of example only, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0211] In some embodiments, once improvement of the patient's symptoms / disorder / condition has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, can be reduced, as a ftmction of the symptoms, to a level at which the improved disease, disorder or condition is retained. Patients can, however, require intermitent treatment on a long-term basis upon any recurrence of symptoms.

[0212] In some embodiments, the amount of a given agent that will correspond to such an amount will vary depending upon factors such as the particular compound, disease and itsAtorney Docket No. UM-43572.601

[0213] Client Ref. No. UM 2025-153 severity, the identity (e.g., weight) of the subject or host in need of treatment, but can nevertheless be determined in a manner recognized in the field according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the route of administration, the condition being treated, and the subject or host being treated. In general, however, doses employed for adult human treatment will typically be in the range of about 0.02 - about 5000 mg per day, in some embodiments, about 1 - about 1500 mg per day. The desired dose may conveniently be presented in a single dose or as divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example as two, three, four or more sub-doses per day.

[0214] As discussed above, provided in certain embodiments herein are combination therapies in which a MC4R agonist peptide is co-administered with an additional agent for the treatment of the disorder / condition, a side effect of the primary agent, or a comorbidity of the disorder / condition. Co-administered agents do not have to be administered in the same pharmaceutical composition, and may, because of different physical and chemical characteristics, have to be administered by different routes. Co-administered agents may be administered concurrently (in the same or separate formulations / compositions) or at separate times (separated by minutes, hours, days, etc.) The co-administered agents may be administered concurrently (e.g., simultaneously, essentially simultaneously or within the same treatment protocol) or sequentially, depending upon the nature of the disease, disorder, or condition, the condition of the patient, and the actual choice of agent used. The determination of the order of administration, and the number of repetitions of administration of each therapeutic agent during a treatment protocol, is well within the knowledge of the clinician after evaluation of the disease being treated and the condition of the patient.

[0215] Therapeutically-effective dosages can vary when the drugs are used in treatment combinations. Methods for experimentally determining therapeutically-effective dosages of drugs and other agents for use in combination treatment regimens are described in the literature. For example, the use of metronomic dosing, i.e., providing more frequent, lower doses in order to minimize toxic side effects, has been described extensively in the literature. Combination treatment fiirther includes periodic treatments that start and stop at various times to assist with the clinical management of the patient.

[0216] For combination therapies described herein, dosages of the co-administered agents will of course vary depending on the type of co-drug employed, on the specific drug employed, on the disease being treated and so forth. In addition, when co-administered withAtorney Docket No. UM-43572.601

[0217] Client Ref. No. UM 2025-153 one or more biologically active agents, the compound provided herein may be administered either simultaneously with the biologically active agent(s), or sequentially.

[0218] In some embodiments, MC4R activation hypersensitizes a subject to the effects (e.g., anorectic effects) of a co-administered anti-obesity agent. In some embodiments, the hypersensitization results in enhanced efficacy of the co-administered anti-obesity agent, thereby increasing the effect (e.g., beyond the therapeutic effect of the MC4R activation alone, beyond the therapeutic effect of the anti-obesity agent alone, beyond the additive therapeutic effect of the MC4R activation and anti-obesity agent), and increasing the effectiveness of the therapy. In some embodiments, the hypersensitization results in a reduction in the therapeutically effective dose when compared to the therapeutically effective dose for the administration of the anti-obesity agent administered alone. In some embodiments, when co-administered with MC4R activation, an anti-obesity agent is administered at a sub-therapeutic dose. In some embodiments, when co-administered with MC4R activation, an anti-obesity agent is administered at a sub-standard dose. In some embodiments, administering an anti-obesity agent at a sub-therapeutic and / or sub-standard dose (e.g., co-administering with MC4R activation) results in a reduction in side effects associated with the treatment when compared to the side effects associated with the therapeutically effective dose and / or standard dose for administration of the anti-obesity agent alone.

[0219] In some embodiments, standard doses for various therapeutics (e.g., anti-obesity agents) are well understood in the field. In some embodiments, methods are provided comprising activating MC4R activity and administering an anti-obesity agent at a substandard dose (e.g., co-administering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity and a sub-standard dose of an anti-obesity agent. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of an anti-obesity agent. In some embodiments, pharmaceutical compositions are provided comprising an activator of MC4R and a substandard dose of an anti-obesity agent.

[0220] The standard dose for subcutaneous administration of liraglutide is 3 mg per week. In some embodiments, methods are provided comprising inhibiting enhancing MC4R expression or activity and administering liraglutide at a sub-standard dose (e.g., coadministering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R antagonist) and a sub-standard dose of liraglutide. In some embodiments, pharmaceutical compositions areAtorney Docket No. UM-43572.601

[0221] Client Ref. No. UM 2025-153 provided comprising a sub-standard dose of liraglutide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of liraglutide. In some embodiments, a dose of less than 3 mg of liraglutide is administered and / or provided in a pharmaceutical formulation (e.g., 2.5 mg, 2 mg, 1.5 mg, 1 mg, 0.5 mg, 0.1 mg, or less, or ranges therebetween).

[0222] The standard dose for subcutaneous administration of metreleptin is 2.5-10 mg per day. In some embodiments, methods are provided comprising enhancing MC4R activity and administering metreleptin at a sub-standard dose (e.g., co-administering with MC4R activation). In some embodiments, methods are provided comprising co-administering an an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of metreleptin. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of metreleptin. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of metreleptin. In some embodiments, a dose of less than 2.5-10 mg of metreleptin is administered and / or provided in a pharmaceutical formulation (e.g., 7.5 mg, 5.0 mg, 2.5 mg, 2 mg, 1.5 mg, 1 mg, 0.5 mg, 0.1 mg, or less, or ranges therebetween).

[0223] The standard dose for oral administration of orlistat is 60 mg (over-the-counter dose) or 120 mg (prescription dose) with each meal. In some embodiments, methods are provided comprising enhancing MC4R activity) and administering orlistat at a sub-standard dose (e.g., co-administering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of orlistat. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of orlistat. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of orlistat. In some embodiments, a dose of less than 60 mg of orlistat is administered and / or provided in a pharmaceutical formulation (e.g., 50 mg, 40 mg, 30 mg, 20 mg, 10 mg, 5 mg, 2 mg, 1 mg, or less, or ranges therebetween).

[0224] The standard dose for oral administration of phentermine and topiramate coadministration is 3.75-15 mg of phentermine (e.g., 3.75 mg, 7.5 mg, 11.25 mg, 15 mg, etc.) and 23-92 mg of topiramate (e.g., 23 mg, 46 mg, 69 mg, 92 mg, etc.) per day. In some embodiments, methods are provided comprising enhancing MC4R activity and administering phentermine and topiramate at a sub-standard dose (e.g., co-administering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of phentermineAtorney Docket No. UM-43572.601

[0225] Client Ref. No. UM 2025-153 and topiramate. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of phentermine and topiramate. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of phentermine and topiramate. In some embodiments, a dose of less than 3.75-15 mg of phentermine (e.g., 10 mg, 7.5 mg, 5 mg , 4 mg, 3 mg, 2 mg, 1 mg, 0.5 mg, or less, or ranges therebetween) and / or less than 23-92 mg of topiramate (e.g., 80 mg, 60 mg, 40 mg , 20 mg, 10 mg, 5 mg, 2 mg, 1 mg, or less, or ranges therebetween) is administered and / or provided in a pharmaceutical formulation.

[0226] The standard dose for oral administration of naltrexone / bupropion is 90-360 mg per day. In some embodiments, methods are provided comprising enhancing MC4R activity and administering naltrexone / bupropion at a sub-standard dose (e.g., co-administering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of naltrexone / bupropion. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of naltrexone / bupropion. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of naltrexone / bupropion. In some embodiments, a dose of less than 90-360 mg of naltrexone / bupropion is administered and / or provided in a pharmaceutical formulation (e.g., 300 mg, 250 mg, 200 mg, 150 mg, 100 mg, 75 mg, 50 mg, 25 mg, 10 mg, or less, or ranges therebetween).

[0227] The standard dose for subcutaneous administration of semaglutide for the treatment of obesity is 0.25-2.0 mg per week. In some embodiments, methods are provided comprising enhancing MC4R activity and administering semaglutide at a sub-standard dose (e.g., coadministering with MC4R activation). In some embodiments, methods are provided comprising co-administering an inh activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of semaglutide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of semaglutide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of semaglutide. In some embodiments, a dose of less than 0.25-2.0 mg of semaglutide is administered and / or provided in a pharmaceutical formulation (e.g., 1.5 mg, 1.0 mg, 0.5 mg 0.2 mg, 0.15 mg, 0.1 mg , 0.05 mg, or less, or ranges therebetween).

[0228] The standard dose for subcutaneous administration of setmelanotide for the treatment of obesity is 1-3 mg per week. In some embodiments, methods are provided comprising enhancing MC4R activity and administering setmelanotide at a sub-standard dose (e.g., co-Atorney Docket No. UM-43572.601

[0229] Client Ref. No. UM 2025-153 administering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of setmelanotide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of setmelanotide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of setmelanotide. In some embodiments, a dose of less than 1-3 mg of setmelanotide is administered and / or provided in a pharmaceutical formulation (e.g., 2.5 mg, 2 mg, 1.5 mg , 1 mg, 0.5 mg, 0.3 mg, 0.2 mg, 0.1 mg, or less, or ranges therebetween).

[0230] The standard dose for subcutaneous administration of tirzepatide for the treatment of obesity is 2.5-15 mg per week. In some embodiments, methods are provided comprising enhancing MC4R activity and administering tirzepatide at a sub-standard dose (e.g., coadministering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of tirzepatide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of tirzepatide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of tirzepatide. In some embodiments, a dose of less than 2.5-15 mg of tirzepatide is administered and / or provided in a pharmaceutical formulation (e.g., 10 mg, 7.5 mg, 5 mg, 2.5 mg, 2 mg, 1.5 mg, 1 mg, 0.5 mg, or less, or ranges therebetween).

[0231] The standard dose for subcutaneous administration of leptin for the treatment of obesity is 2.5-10 mg per day. In some embodiments, methods are provided comprising enhancing MC4R activity and administering leptin at a sub-standard dose (e.g., coadministering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of leptin. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of leptin. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of leptin. In some embodiments, a dose of less than 2.5-15 mg of leptin is administered and / or provided in a pharmaceutical formulation (e.g., 7.5 mg, 5 mg, 2.5 mg, 2 mg, 1.5 mg, 1 mg, 0.5 mg, or less, or ranges therebetween).

[0232] The standard dose for subcutaneous administration of exendin-4 or exenatide for the treatment of obesity is 2 mg per week. In some embodiments, methods are provided comprising enhancing MC4R activity and administering exendin-4 or exenatide at a substandard dose (e.g., co-administering with MC4R activation). In some embodiments, methodsAtorney Docket No. UM-43572.601

[0233] Client Ref. No. UM 2025-153 are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of exendin-4 or exenatide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of exendin-4 or exenatide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of exendin-4 or exenatide. In some embodiments, a dose of less than 2 mg of exendin-4 or exenatide is administered and / or provided in a pharmaceutical formulation (e.g., 1.5 mg, 1 mg, 0.5 mg, 0.1 mg, or less, or ranges therebetween).

[0234] The standard dose for subcutaneous administration of albiglutide for the treatment of obesity is 30 mg per week. In some embodiments, methods are provided comprising enhancing MC4R activity) and administering albiglutide at a sub-standard dose (e.g., coadministering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of albiglutide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of albiglutide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of albiglutide. In some embodiments, a dose of less than 30 mg of albiglutide is administered and / or provided in a pharmaceutical formulation (e.g., 25 mg, 20 mg, 15 mg, 10 mg, 5 mg, 1 mg, 0.5 mg, or less, or ranges therebetween).

[0235] The standard dose for subcutaneous administration of dulaglutide for the treatment of obesity is 0.75-4.5 mg per week. In some embodiments, methods are provided comprising enhancing MC4R activity and administering dulaglutide at a sub-standard dose (e.g., coadministering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of dulaglutide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of dulaglutide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of dulaglutide. In some embodiments, a dose of less than 0.75-4.5 mg of dulaglutide is administered and / or provided in a pharmaceutical formulation (e.g., 4 mg, 2 mg , 1 mg, 0.5 mg, 0.2 mg, 0.1 mg, or less, or ranges therebetween).

[0236] The standard dose for subcutaneous administration of lixisenatide for the treatment of obesity is 10-20 pg per day. In some embodiments, methods are provided comprising enhancing MC4R activity and administering lixisenatide at a sub-standard dose (e.g., coadministering with MC4R activation). In some embodiments, methods are providedAtorney Docket No. UM-43572.601

[0237] Client Ref. No. UM 2025-153 comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of lixisenatide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of lixisenatide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of lixisenatide. In some embodiments, a dose of less than 10-20 mg of lixisenatide is administered and / or provided in a pharmaceutical formulation (e.g., 15 mg, 10 mg, 5 mg, 1 mg, 0.5 mg, or less, or ranges therebetween).

[0238] The standard dose for subcutaneous administration of taspoglutide is 10-20 pg per day. In some embodiments, methods are provided comprising enhancing MC4R expression or activity and administering taspoglutide at a sub-standard dose (e.g., co-administering with MC4R activation). In some embodiments, methods are provided comprising co-administering an activator of MC4R activity (e.g., an MC4R agonist) and a sub-standard dose of taspoglutide. In some embodiments, pharmaceutical compositions are provided comprising a sub-standard dose of taspoglutide. In some embodiments, pharmaceutical compositions are provided comprising an MC4R activator (e.g., agonist) and a sub-standard dose of taspoglutide. In some embodiments, a dose of less than 10-20 mg of taspoglutide is administered and / or provided in a pharmaceutical formulation (e.g., 15 mg, 10 mg, 5 mg, 1 mg, 0.5 mg, or less, or ranges therebetween).

[0239] In some embodiments, a sub-standard dose is less than 10-90% of the standard dose (e.g., <10%, <20%, <30%, <40%, <50%, <60%, <70%, <80%, <90%, etc.).

[0240] EXPERIMENTAL

[0241] Example 1

[0242] Pharmacological in vitro Assays

[0243] Determination of intracellular cAMP levels in live cells: A genetically-encoded cAMP split-luciferase reporter stably-expressing cell line (Promega, Madison, WI)(Binkowski et al., 2011, ACS chemical biology 6, 1193-1197.; incorporated by reference in its entirety) was used as the basis for the generation of stable clones expressing the human MC4R receptor (a gift from Promega), or the human MC3R (generated in-house by clonal selection). The stable cell lines were grown and maintained in selection media consisting of Dulbecco’s modified Eagle media (DMEM) with 4.5 g / 1 D-glucose, and 4 mM L-glutamine (Thermo Fisher Scientific, Waltham, MA), supplemented with 10% fetal bovine serum, 100 units / ml penicillin, 100 pg / ml streptomycin, 2.5 pg / ml amphotericin B, 200 pg / ml hygromycin B (for positive selection of the GScAMP22f luciferase reporter), andAtorney Docket No. UM-43572.601

[0244] Client Ref. No. UM 2025-153 Geneticin™ (G418) 700 pg / ml (for MC3R or MC4R selection). Actual serum concentration during the assay is estimated to be about 1%. Cell line identity is routinely verified by qPCR and MC1R-, MC5R-, MC3R- and MC4R-specific oligonucleotides.

[0245] The assay for the determination of the cAMP response in live cells was described previously. (Yu et al., 2020, Science 368, 428-433; incorporated by reference in its entirety). Cells were seeded at a density of 20,000 cells per well using 384-well poly-D lysine-coated, clear botom, and black-wall assay plates (Coming Inc. Coming, NJ). Cells were allowed to atach to the plates for 18 to 24 h after which growth media was removed and 20 pl of 4% D-luciferin (Promega) in CO2 independent, serum-free medium (Thermo Fisher Scientific) was added to each well. The luciferase substrate was allowed to permeate the cells for 120 min at 37°C. Intracellular cAMP levels were measured using an FDSS 7000EX Functional Drug Screening System (Hamamatsu Photonics, Hamamatsu, Japan) in the Center for Chemical Genomics at the Life Sciences Institute. This instrument allowed the in-line addition of test-peptides and receptor agonists while simultaneously acquiring the luminescence signal from live cells. Assay read steps were set as follows: baseline acquisition of 2 min, the addition of 10 pl of varying 3x concentrations of test-peptides or vehicle followed by 11 min measurement (measurement window 1), and 10 pl addition of 4x concentration of the endogenous melanocortin agonist a-MSH (Bachem, Bubendorf, Switzerland) followed by an additional 11 min response measurement (measurement window 2). The resulting final concentration of a-MSH was close to the respective receptor EC90 dose for each receptor. Intraplate concentration response curves for a-MSH and SHU-9119 (Phoenix Pharmaceuticals, Burlingame CA) were included as reference controls. A submaximal forskolin (20 pM) concentration was also included to serve as a normalization reference to account for cell number variations and differences in assay transducer efficiency between cell lines.

[0246] With this set up it was possible to evaluate the direct effect of the test-peptides on the MC1R, MC5R, MC3R and MC4R cell lines during measurement window 1, while the antagonist profile in the presence of EC90 a-MSH was determined on measurement window 2. For data analysis baseline luminescence (i.e. the maximum luminescence signal from the initial 0 to 2 min window) was subtracted from the maximum luminescence obtained during measurement window 1 (2 to 13 min) and measurement window 2 (13 to 24 min) to yield the test-peptide elicited responses. EC50 or IC50 potency values were determined by non-linear regression by fiting the data to a sigmoid four-parameter variable slope model using the GraphPad Prism version 8.4 software package (San Diego CA).Attorney Docket No. UM-43572.601

[0247] Client Ref. No. UM 2025-153 Exemplary results of pharmacological in vitro assays are provided in Tables 1-2.Attorney Docket No. UM-43572.601

[0248] Client Ref. No. UM 2025-153

[0249] Table 1. MC4R cAMP (EC50) and efficacy (%) for exemplary MC4R agonists.

[0250]

[0251] Atorney Docket No. UM-43572.601

[0252] Client Ref. No. UM 2025-153

[0253]

[0254] Atorney Docket No. UM-43572.601

[0255] Client Ref. No. UM 2025-153

[0256]

[0257] Atorney Docket No. UM-43572.601

[0258] Client Ref. No. UM 2025-153

[0259]

[0260] Attorney Docket No. UM-43572.601

[0261] Client Ref. No. UM 2025-153

[0262] Table 2. Potency-based MC4R selectivity versus MC3R and MC1R for exemplary MC4R agonists

[0263]

[0264] Atorney Docket No. UM-43572.601

[0265] Client Ref. No. UM 2025-153

[0266]

[0267] Atorney Docket No. UM-43572.601

[0268] Client Ref. No. UM 2025-153

[0269]

[0270] Atorney Docket No. UM-43572.601

[0271] Client Ref. No. UM 2025-153

[0272]

[0273] Atorney Docket No. UM-43572.601

[0274] Client Ref. No. UM 2025-153 Example 2

[0275] Experiments conducted during development of embodiments herein have demonstrated favorable characteristics of multiple substitutions exemplified in the peptides herein, for example:

[0276] • Cit substitution for His at AA4 increases MC4R selectivity over MC1R;

[0277] • Wpb substitution for Trp at AA7 increases MC4R potency;

[0278] • Cit, hCit, Pal(2’), Pal(3’), Pal(4‘) substitution for His at AA4 of peptides cyclized from AA2 to AA8 increases MC4R selectivity;

[0279] • Wpb substitution for Trp at AA7 and Cit / Alb substitution for His at AA4 of peptides cyclized from AA2 to AA8 increases MC4R selectivity; and

[0280] • Cit substitution for His and AA4 and Wpb substitutions for Trp at AA7 of peptides cyclized from AA3 to AA8 increases MC4R selectivity.

[0281] Example 3

[0282] Experiments conducted during development of embodiments herein have demonstrated that peptides described herein and previously (e.g., CTX-1715 andCTX-1718; CTX-1228 (Arg-Nle-cycZo(Cys-Arg-DPhe-Arg-Trp-Cys); SEQ ID NO: 112); and CTX-1649 (cyclo[S-acetyl-Aib-Cit-DPhe-Arg-Trp-Cys]-NH2); SEQ ID NO: 113) outperform Setmelanotide in synergizing with Semaglutide to induce weight loss. CTX-1715, CTX-1718, and CTX-1649 are also differentiated from Setmelanotide in being MClR-sparing, as defined by a MC4R / MC1R ratio >100 and / or an MC1R Emax<25%. In the 24 hour acute feeding assay in WT mice on normal chow, Setmelanotide and peptides described herein both showed an ability to synergize with Semaglutide at subtherapeutic doses. In this chronic assay, Setmelanotide shows no ability to synergize with Semaglutide-induced weight loss. CTX-1649 also demonstrates potent synergistic weight loss with Semaglutide at subtherapeutic doses out to 10 days of administration, further differentiating this compound from Setmelanotide. Thus, CTX-1649 is differentiated from Setmelanotide in three characteristics, 1) MClR-sparing, 2) synergy with Semaglutide in magnitude of weight loss, and 3) ability to act at true subtherapeutic doses.

[0283] Example 4

[0284] Synergistic weight lossAtorney Docket No. UM-43572.601

[0285] Client Ref. No. UM 2025-153 14-week-old male WT C57BL / 6J DIO mice were obtained from the Jackson Laboratory (Cat. # 380050). Mice were single-housed and maintained on high-fat chow (Research Diets #12492, 60 kcal% fat, 5.2 kcal / gram) under a 12hr:12hr lightdark cycle throughout the experiment. Upon reaching 35-40 grams of body weight, mice were randomized into the treatment groups shown in each experiment. Before commencing treatments, mice were acclimated to daily subcutaneous injections of sterile saline (100-200 pL) for 5 days prior to experimentation. Mice were then treated once daily with subcutaneous administration of tirzepatide (2nmol / kg), cagrilintide (lOnmol / kg), tirzepatide (2nmol / kg) plus CTX-1649 (Img / kg), or cagrilintide (lOnmol / kg) plus CTX-1649 (Img / kg) for the length of time indicated. In the tirzepatide trial (left panel), drug treatment was terminated, and weight regain was observed (pink section labelled “reb”). Data (Figure 4) show %change in body weigh from day 0, N-7 per group.

[0286] Example 5

[0287] Delayed weight regain after cessation of treatment

[0288] 14-week-old male WT C57BL / 6J DIO mice were obtained from the Jackson Laboratory (Cat. # 380050). Mice were single-housed and maintained on high-fat chow (Research Diets #12492, 60 kcal% fat, 5.2 kcal / gram) under a 12hr:12hr lightdark cycle throughout the experiment. Upon reaching 35-40 grams of body weight, mice were randomized into the treatment groups shown in each experiment. Before commencing treatments, mice were acclimated to daily subcutaneous injections of sterile saline (100-200 pL) for 5 days prior to experimentation. Mice were then treated once daily with subcutaneous administration of saline, semaglutide (lOnmol / kg), setmelanotide, CTX-1715, or CTX-1649 (Img / kg), or semaglutide (lOnmol / kg) plus the melanocortin peptide indicated (Img / kg) for the length of time indicated. The same saline and semaglutide curves are reproduced in each panel. The left panel indicates effects of treatment with setmelanotide or setmelanotide plus semaglutide. The middle panel indicates effects of treatment with CTX-1715 or CTX-1715 plus semaglutide. And the right panel indicates the effect of treatment of CTX-1649, or CTX-1649 plus semaglutide. Gray shading indicates the treatment period, the white area indicates the weight regain following removal of all drug treatment. Data show %change in body weigh from day 0, N-7 per group. The delayed weight regain after combination treatment can also be seen for tirzepatide plus CTX-1649 in Figure 5.

[0289] Example 6Atorney Docket No. UM-43572.601

[0290] Client Ref. No. UM 2025-153 Combination therapy results in proportionately more fat mass loss rather than lean mass loss

[0291] Combination therapy with CTX-melanocortin peptides preferentially increases fat mass loss (Figure 6). Combination treatment with tirzepatide and CTX-1649 doubled fat mass loss (middle panel) compared to tirzepatide alone, with no increasing loss of lean mass (right panel). N=5.

[0292] Example 7

[0293] CTX Peptides show significant improvement in the treatment of MC4R haploinsufficiency, relative to setmelanotide (Figure 7). A subset of peptides were tested for their ability to inhibit food intake in a 24hr feeding assay in the MC4R+ / - mouse (Fig. 7, top panel). Several peptides show greatly enhanced in vivo potency. Many peptides also exhibited significant improvement in MC4R selectivity in this assay (blue bars) relative to setmelanotide (red bar). Increased ratio of MC4R potency to MC1R potency is shown in green bars (log scale). One exemplary compound, CTX-1715, was then tested in a 14 day weight loss assay (Fig. 7, botom panel). Using obese male MC4R+ / - mice, animals were treated daily with setmelanotide or CTX-1715 (each at Img / kg). Setmelanotide produced -2.5% weight loss, while CTX-15 yielded 12.5-15% weight loss. N=7 / group. Fat mass loss, determined by NMR.

[0294] Example 8

[0295] Cardiovascular safety

[0296] Setmelanotide demonstrates no cardiovascular effects in humans at therapeutic doses. Using an SC dose of Img / kg in instrumented Rhesus Group average of 5 animals over each of 3 days of treatment @ Img / mL, CTX-1715 shows similar or fewer CV effects than Setmelanotide at equivalent doses in NHP (Figure 8). CTX-1649 similarly shows no cardiovascular effects in NHP.

[0297] Example 9

[0298] Oral Bioavailability

[0299] CTX-1649 and CTX-1715 can be detected in plasma after oral gavage in the mouse (Figure 9).Atorney Docket No. UM-43572.601

[0300] Client Ref. No. UM 2025-153 SEQUENCES

[0301] In the following sequences, “c[” indicates the beginning of a cyclic portion and “]” indicates the end of a cyclic portion. As depicted in Figure 3, the first amino acid following the “c[” is linked to the last amino acid prior to the “]”. Peptides comprising the following sequences (or having a degree of sequence identity thereto) may farther comprise N-terminal or C-terminal caps or modifications, such as the peptides listed in Tables 1 and 2.

[0302] SEQ IDNO: 1 X-AA0-AAl-AAlB-c[AA2-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is acetyl, 4-I-phenyl-(CH2)3-C[=O] (or 4-I-phenyl with an alternative linker), R1 as disclosed herein, or absent;

[0303] wherein AAO is absent or bAla;

[0304] wherein AA1 is Arg or absent;

[0305] wherein AA1B is Nle, Phe, Hph, Gly, Ala, D-Ala, or absent;

[0306] wherein AA2 is Cys;

[0307] wherein AA3 is D-Ala, Pro, D-Nle, D-Hle, Aib, Gly, beta-Ala, D-Alb, or absent;

[0308] wherein AA4 is His, Alb, Nar, Arg, Cit, Om, Pal(4’), Pal(2’), Pal(3’), or homoCit;

[0309] wherein AA5 is D-Phe, D-Fpy, D-Fpm, D-Fpt, D-Tyr(4-OMe), or Phe; wherein AA6 is Arg;

[0310] wherein AA7 is Trp, Wpb, or Phe;

[0311] wherein AA8 is Cys;

[0312] wherein AA9 is Nva, Hie, Cpa, Phg, Abu, Om, D-Hph, or absent; wherein AA1 is linked to AA2 if AA1B is absent; and

[0313] wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent.

[0314] SEQ ID NO: 2 X-AA0-AAl-AAlB-c[AA2-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is acetyl, 4-I-phenyl-(CH2)3-C[=O] (or 4-I-phenyl with an alternative linker), R1 as disclosed herein, or absent;

[0315] wherein AAO is absent or bAla;

[0316] wherein AA1 is Arg or absent;Atorney Docket No. UM-43572.601

[0317] Client Ref. No. UM 2025-153 wherein AA1B is Nle, Phe, Hph, Gly, Ala, D-Ala, or absent;

[0318] wherein AA2 is Cys;

[0319] wherein AA3 is D-Ala, Pro, D-Nle, D-Hle, Aib, or absent;

[0320] wherein AA4 is His, Alb, Nar, Arg, Cit, Om, Pal(4’);

[0321] wherein AA5 is D-Phe, D-Fpy, D-Fpm, D-Fpt;

[0322] wherein AA6 is Arg;

[0323] wherein AA7 is Trp or Wpb;

[0324] wherein AA8 is Cys;

[0325] wherein AA9 is Hie, Cpa, Phg, Abu, Om, D-Hph, or absent; and wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent

[0326] SEQ ID NO: 3 (e.g., CTX-1659) - Phe c[Cys DAla His DPhe Arg Trp Cys]

[0327] SEQ ID NO: 4 (e.g., CTX-1660) - Hph c[Cys DAla His DPhe Arg Trp Cys]

[0328] SEQ ID NO: 5 (e.g., CTX-1661) - Arg c[Cys Pro His DPhe Arg Trp Cys]

[0329] SEQ ID NO: 6 (e.g., CTX-1662) - Arg c[Cys DNle His DPhe Arg Trp Cys]

[0330] SEQ ID NO: 7 (e.g., CTX-1663) - Arg c[Cys DHle His DPhe Arg Trp Cys]

[0331] SEQ ID NO: 8 (e.g., CTX-1665) - Arg c[Cys DAla Nar DPhe Arg Trp Cys]

[0332] SEQ ID NO: 9 (e.g., CTX-1666) - Arg c[Cys DAla Alb DPhe Arg Trp Cys]

[0333] SEQ ID NO: 10 (e.g., CTX-1667) - Arg c[Cys DAla His DFpy Arg Trp Cys]

[0334] SEQ ID NO: 11 (e.g., CTX-1668) - Arg c[Cys DAla His DFpt Arg Trp Cys]

[0335] SEQ ID NO: 12 (e.g., CTX-1669) - Arg c[Cys DAla His DFpm Arg Trp Cys]

[0336] SEQ ID NO: 13 (e.g., CTX-1673) - Arg c[Cys DAla His DPhe Arg Wpb Cys]Atorney Docket No. UM-43572.601

[0337] Client Ref. No. UM 2025-153 SEQ ID NO: 14 (e.g., CTX-1674) - Arg c[Cys DAla His DPhe Arg Trp Cys] Abu

[0338] SEQ ID NO: 15 (e.g., CTX-1675) - Arg c[Cys DAla His DPhe Arg Trp Cys] Hie

[0339] SEQ ID NO: 16 (e.g., CTX-1676) - Arg c[Cys DAla Arg DPhe Arg Trp Cys] Hie

[0340] SEQ ID NO: 17 (e.g., CTX-1677) - Arg c[Cys DAla Cit DPhe Arg Trp Cys] Hie

[0341] SEQ ID NO: 18 (e.g., CTX-1678) - Arg c[Cys DAla His DPhe Arg Trp Cys] Cpa

[0342] SEQ ID NO: 19 (e.g., CTX-1679) - Arg c[Cys DAla His DPhe Arg Trp Cys] Phg

[0343] SEQ ID NO: 20 (e.g., CTX-1680) - Arg c[Cys DAla His DPhe Arg Trp Cys] DHph

[0344] SEQ ID NO: 21 (e.g., CTX-1682) - Arg Nle c[Cys Nar DPhe Arg Trp Cys]

[0345] SEQ ID NO: 22 (e.g., CTX-1683) - Arg Nle c[Cys Alb DPhe Arg Trp Cys]

[0346] SEQ ID NO: 23 (e.g., CTX-1687) - c[S-acetyl DNle His DPhe Arg Trp Cys]

[0347] SEQ ID NO: 24 (e.g., CTX-1688) - c[S-acetyl Dalb His DPhe Arg Trp Cys]

[0348] SEQ ID NO: 25 (e.g., CTX-1689) - c[S-acetyl beta-Ala His DPhe Arg Trp Cys]

[0349] SEQ ID NO: 26 (e.g., CTX-1690) - c[S-acetyl beta-Ala Arg DPhe Arg Trp Cys]

[0350] SEQ ID NO: 27 (e.g., CTX-1691) - c[S-acetyl DAla Arg DPhe Arg Trp Cys] Nva

[0351] SEQ ID NO: 28 (e.g., CTX-1692) - c[S-acetyl DAla Arg DPhe Arg Trp Cys] Hie

[0352] SEQ ID NO: 29 (e.g., CTX-1693) - c[S-acetyl DAla Arg DPhe Arg Trp Cys] Cpa

[0353] SEQ ID NO: 30 (e.g., CTX-1694) - c[S-acetyl DAla Arg DPhe Arg Trp Cys] PhgAtorney Docket No. UM-43572.601

[0354] Client Ref. No. UM 2025-153 SEQ ID NO: 31 (e.g., CTX-1695) - c[S-acetyl beta-Ala Cit DPhe Arg Trp Cys]

[0355] SEQ ID NO: 32 (e.g., CTX-1696) - c[S-acetyl DNle Cit DPhe Arg Trp Cys]

[0356] SEQ ID NO: 33 (e.g., CTX-1697) - c[S-acetyl DAlb Cit DPhe Arg Trp Cys]

[0357] SEQ ID NO: 34 (e.g., CTX-1698) - c[S-acetyl Gly Cit DPhe Arg Trp Cys]

[0358] SEQ ID NO: 35 (e.g., CTX-1699) - c[S-acetyl Gly Cit DPhe Arg Trp Cys]

[0359] SEQ ID NO: 36 (e.g., CTX-1700) - c[S-acetyl Gly Cit DPhe Arg Trp Cys] Hie

[0360] SEQ ID NO: 37 (e.g., CTX-1701) - c[S-acetyl Gly Cit DPhe Arg Trp Cys] Cpa

[0361] SEQ ID NO: 38 (e.g., CTX-1702) - c[S-acetyl Gly Cit DPhe Arg Trp Cys] Phg

[0362] SEQ ID NO: 39 (e.g., CTX-1708) - c[S-acetyl Gly Nar DPhe Arg Trp Cys]

[0363] SEQ ID NO: 40 (e.g., CTX-1709) - c[S-acetyl Gly His DFpy Arg Trp Cys]

[0364] SEQ ID NO: 41 (e.g., CTX-1710) - c[S-acetyl Gly His DPhe Arg Wpb Cys]

[0365] SEQ ID NO: 42 (e.g., CTX-1711) - Arg c[Cys Aib Arg DPhe Arg Wpb Cys]

[0366] SEQ ID NO: 43 (e.g., CTX-1712) - Arg c[Gly Cys Aib Arg DPhe Arg Wpb Cys]

[0367] SEQ ID NO: 44 (e.g., CTX-1713) - Arg c[Ala Cys Aib Arg DPhe Arg Wpb Cys]

[0368] SEQ ID NO: 45 (e.g., CTX-1714) - Arg c[DAla Cys Aib Arg DPhe Arg Wpb Cys]

[0369] SEQ ID NO: 46 (e.g., CTX-1715) - Arg c[Cys Aib Arg DPhe Arg Wpb Cys] Abu

[0370] SEQ ID NO: 47 (e.g., CTX-1716) - Arg c[Cys Aib Nar DPhe Arg Wpb Cys]Atorney Docket No. UM-43572.601

[0371] Client Ref. No. UM 2025-153 SEQ ID NO: 48 (e.g., CTX-1717) - Arg c[Cys Aib Nar DPhe Arg Wpb Cys] Abu

[0372] SEQ ID NO: 49 (e.g., CTX-1718) - Arg c[Cys Aib Cit DPhe Arg Wpb Cys]

[0373] SEQ ID NO: 50 (e.g., CTX-1719) - c[Cys Aib Alb DPhe Arg Wpb Cys]

[0374] SEQ ID NO: 51 (e.g., CTX-1720) - c[Arg Cys Aib Nar DFpy Arg Trp Cys]

[0375] SEQ ID NO: 52 (e.g., CTX-1721) - bAla Arg c[Cys Aib Nar DPhe Arg Wpb Cys]

[0376] SEQ ID NO: 53 (e.g., CTX-1722) - Arg c[Cys Aib Nar DFpy Arg Wpb Cys]

[0377] SEQ ID NO: 54 (e.g., CTX-1723) - Arg c[Cys Aib Alb DFpy Arg Trp Cys]

[0378] SEQ ID NO: 55 (e.g., CTX-1724) - bAla c[Arg Cys Aib Arg DPhe Arg Wpb Cys]

[0379] SEQ ID NO: 56 (e.g., CTX-1726) - Arg c[Cys Aib Cit DFyp Arg Trp Cys]

[0380] SEQ ID NO: 57 (e.g., CTX-1727) - bAla Arg c[Cys Aib Cit DPhe Arg Wpb Cys]

[0381] SEQ ID NO: 58 (e.g., CTX-1729) - Arg Nle c[Cys Om DFpy Arg Trp Cys]

[0382] SEQ ID NO: 59 (e.g., CTX-1730) - Arg Nle c[Cys Om DPhe Arg Wpb Cys]

[0383] SEQ ID NO: 60 (e.g., CTX-1732) - Arg Nle c[Cys Arg DFpy Arg Trp Cys]

[0384] SEQ ID NO: 61 (e.g., CTX-1733) - Arg Nle c[Cys Arg DPhe Arg Wpb Cys]

[0385] SEQ ID NO: 62 (e.g., CTX-1735) - Arg Nle c[Cys Nar DFpy Arg Trp Cys]

[0386] SEQ ID NO: 63 (e.g., CTX-1736) - Arg Nle c[Cys Nar DPhe Arg Wpb Cys]

[0387] SEQ ID NO: 64 (e.g., CTX-1737) - Arg Nle c[Cys Nar DFpy Arg Wpb Cys]Atorney Docket No. UM-43572.601

[0388] Client Ref. No. UM 2025-153 SEQ ID NO: 65 (e.g., CTX-1738) - Arg Nle c[Cys His DFpy Arg Trp Cys]

[0389] SEQ ID NO: 66 (e.g., CTX-1739) - Arg Nle c[Cys His DPhe Arg Wpb Cys]

[0390] SEQ ID NO: 67 (e.g., CTX-1741) - Arg Nle c[Cys Alb DFpy Arg Trp Cys]

[0391] SEQ ID NO: 68 (e.g., CTX-1742) - Arg Nle c[Cys Alb DPhe Arg Wpb Cys]

[0392] SEQ ID NO: 69 (e.g., CTX-1743) - Arg Nle c[Cys Alb DFpy Arg Wpb Cys]

[0393] SEQ ID NO: 70 (e.g., CTX-1744) - Arg Nle c[Cys Arg DPhe Arg Wpb Cys] Abu

[0394] SEQ ID NO: 71 (e.g., CTX-1745) - Arg Nle c[Cys Cit DPhe Arg Wpb Cys] Om

[0395] SEQ ID NO: 72 (e.g., CTX-1746) - c[S-acetyl Gly His DFpy Arg Wpb Cys]

[0396] SEQ ID NO: 73 (e.g., CTX-1747) - c[S-acetyl Gly Alb DPhe Arg Wpb Cys]

[0397] SEQ ID NO: 74 (e.g., CTX-1749) - c[S-acetyl Gly Cit DPhe Arg Wpb Cys]

[0398] SEQ ID NO: 75 (e.g., CTX-1751) - c[S-acetyl Gly Arg DPhe Arg Wpb Cys]

[0399] SEQ ID NO: 76 (e.g., CTX-1753) - c[S-acetyl bAla His DFpy Arg Wpb Cys]

[0400] SEQ ID NO: 77 (e.g., CTX-1754) - c[S-acetyl bAla His DPhe Arg Wpb Cys]

[0401] SEQ ID NO: 78 (e.g., CTX-1755) - c[S-acetyl bAla Arg DPhe Arg Wpb Cys]

[0402] SEQ ID NO: 79 (e.g., CTX-1756) - c[S-acetyl bAla Arg DPhe Arg Wpb Cys] Om

[0403] SEQ ID NO: 80 (e.g., CTX-1757) - c[S-acetyl bAla Arg DPhe Arg Wpb Cys] Abu

[0404] SEQ ID NO: 81 (e.g., CTX-1761) - Arg c[Cys Aib Cit DPhe Arg Wpb Cys]Atorney Docket No. UM-43572.601

[0405] Client Ref. No. UM 2025-153 SEQ ID NO: 82 (e.g., CTX-1763) - Arg c[Cys Aib Pal(4’) DPhe Arg Wpb Cys]

[0406] SEQ ID NO: 83 (e.g., CTX-1764) - Arg c[Cys* Aib Cit DPhe Arg Wpb Cys*]

[0407] SEQ ID NO: 84 (e.g., CTX-1765) - Arg Nle c[Cys Cit DPhe Arg Wpb Cys]

[0408] SEQ ID NO: 85 (e.g., CTX-1766) - Arg Nle c[Cys Pal(4’) DPhe Arg Wpb Cys]

[0409] SEQ ID NO: 86 (e.g., CTX-1770) - Arg Nle c[Cys* Cit DPhe Arg Wpb Cys*]

[0410] SEQ ID NO: 87 (e.g., CTX-1771) - c[S-acetyl Aib Cit DPhe Arg Trp Cys Om]

[0411] SEQ ID NO: 88 (e.g., CTX-1772) - c[S-acetyl betaAla Arg DPhe Arg Wpb Cys] Om]

[0412] SEQ ID NO: 89 (e.g., CTX-1773) - c[S-acetyl Gly Cit DPhe Arg Wpb Cys] Om

[0413] SEQ ID NO: 90 c[X-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y;

[0414] wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is S-acetyl;

[0415] wherein AA3 is Aib, D-Ala, Gly, D-Nle, D-Alb, or beta- Ala;

[0416] wherein AA4 is Arg, Cit, His, Nar, Alb, Om, Pal(4’), homoCit, Pal(2’), Pal(3’);

[0417] wherein AA5 is D-Phe, D-Fpy, D-Fpm, D-Fpt, D-Tyr(4-OMe), or Phe; wherein AA6 is Arg;

[0418] wherein AA7 is Trp, Wpb, D-Trp, or Phe;

[0419] wherein AA8 is Cys;

[0420] wherein AA9 is Nva, Hie, Cpa, Phg, Abu, Om, D-Hph, or absent wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent

[0421] SEQ ID NO: 91 c[X-AA3-AA4-AA5-AA6-AA7-AA8]-AA9-Y;

[0422] wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is S-acetyl;Atorney Docket No. UM-43572.601

[0423] Client Ref. No. UM 2025-153 wherein AA3 is Aib, D-Ala, Gly, D-Nle, D-Alb, or beta- Ala;

[0424] wherein AA4 is Arg, Cit, His, or Nar;

[0425] wherein AA5 is D-Phe;

[0426] wherein AA6 is Arg;

[0427] wherein AA7 is Trp or Wpb;

[0428] wherein AA8 is Cys;

[0429] wherein AA9 is Nva, Hie, Cpa, Phg, or absent

[0430] wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent

[0431] SEQ ID NO: 92 X-AA0-AA1-AA1B-AA2-AA3-AA4-AA5-AA6-AA7-AA8-AA9-AA10-AA11-Y;

[0432] wherein X is a N-terminal cap moiety linked to the most N-terminal amino acid of the peptide and is acetyl, chloro acetyl, 4-I-phenyl-(CH2)3-C[=O] (or 4-I-phenyl with an alternative linker), or absent;

[0433] wherein AAO is absent or bAla;

[0434] wherein AA1 is Arg or absent;

[0435] wherein AA1B is Nle, Phe, HpH, Gly, Ala, or absent; wherein AA2 is Cys, hCys, or absent;

[0436] wherein AA3 is D-Ala, Glu, D-Glu, D-Gly, D-Aib, Gly, Ala, NMe-Ala, Aib, Abu, D-Abu, Pro, D-Nle, D-Hle, D-Alb, beta- Ala, or absent;

[0437] wherein AA4 is Arg, D-Arg, NMe-Arg, NMe-D-Arg, Cit, D- Cit, His, D-His, Nme-His, NMe-D-His, Pro, D-Om, Om, Homo- Arg, Homo-Cit, Homo-D-Cit, Pal(2’), Pal(3’), Pal(4’), D-Pal(2’), D-Pal(3’), 4-guanidyl-Dab, 4-guanidyl-D-Dab, 3-guanidyl-Dap, 3-guanidyl-D-Dap, 5-carbamoyl-Dab, 5-carbamoyl-D-Dab, 3-carbamoyl-Dap, 3-carbamoyl-D-Dap, Agp, Nar, Alb, or D-Pal(4’);

[0438] wherein AA5 is Phe, D-Phe, D-Phe(4-Br), D-Phe(4-I), D-Phe(4-F), D-Phe(4-tBu), Phe(4-Br), Phe(4-F), D-Tyr, Tyr(4-OMe), or D-Tyr(4-OMe), D-Hph, D-Bip, D-Tic, D-Dip, D-Trp, aMe-D-Phe, D-Phe(4-NH-Ac), NMe-D-Phe, Phe(4-tBu), Trp, Hph, Bip, Tic, Dip, aMe-Phe, Phe(4-NH-Ac), NMe-Phe, Tyr, Phe(4-I), D-Phe(4-I), Phe(4-tBu)], D-Phe(4-guanidyl), D-Fpy, D-Fpt, D-Fpm, or Phe(4-guanidyl);

[0439] wherein AA6 is Arg, NMe-Arg, D-Arg, Cit, NMe-D-Arg, or D-Cit;Atorney Docket No. UM-43572.601

[0440] Client Ref. No. UM 2025-153 wherein AA7 is Trp, D-Trp, NMe-Trp, Phe, D-Phe, D-Ala D-Tic, NMe-D-Trp, Ala, Wpb, or Tic;

[0441] wherein AA8 is Cys or L-Pen;

[0442] wherein AA9 is Lys, Arg, Abu, Hie, Cpa, Phg, D-Hph, Nva, Om, or absent; wherein AA10 is Pro, Phe, D-Phe, or absent;

[0443] wherein AA11 is Vai, Gly, or absent;

[0444] wherein Y is a C-terminal cap linked to the most C-terminal amino acid of the peptide and is NH2 or absent;

[0445] wherein AA1 is linked to AA2 if AA1B is absent;

[0446] wherein AA9 is present if AA10 is present; wherein AA9 and AA10 are present if AA11 is present;

[0447] wherein the peptide comprises at least one of:

[0448] bAla at AAO;

[0449] Phe, HpH, Gly, Ala at AA1B;

[0450] hCys at AA2;

[0451] Pro, D-Nle, D-Hle, D-Alb, beta- Ala at AA3;

[0452] Agp, Nar, Thr, Cit, or Alb at AA4;

[0453] D-Fpy, D-Fpt, D-Fpm at AA5;

[0454] Wpb at AA7;

[0455] L-Pen at AA8; or

[0456] Abu, Hie, Cpa, Phg, D-Hph, Nva, or Om at AA9.

[0457] SEQ ID NO: 93 (e.g., CTX-1780) - c[S-acetyl-Aib-Cit-DPhe-Arg-Wpb-hCys]

[0458] SEQ ID NO: 94 (e.g., CTX-1781) - c[S-acetyl-Aib-Cit-DPhe-Arg-Wpb-Pen]

[0459] SEQ ID NO: 95 (e.g., CTX-1789) - Ac-Arg-c[Cys*-Aib-Arg-DPhe-Arg-Wpb-Cys*]-Abu

[0460] SEQ ID NO: 96 (e.g., CTX-1790) - Ac-Arg-c[Cys*-Aib-Cit-DPhe-Arg-Wpb-Cys*]-Abu

[0461] SEQ ID NO: 97 (e.g., CTX-1793) - Ac-Arg-c[Cys-Gly-Cit-DPhe-Arg-Wpb-Cys]-Abu

[0462] SEQ ID NO: 98 (e.g., CTX-1794) - Arg-c[Cys-Gly-Cit-DPhe-Arg-Wpb-Cys]Atorney Docket No. UM-43572.601

[0463] Client Ref. No. UM 2025-153 SEQ ID NO: 99 (e.g., CTX-1795) - Arg-c[Cys-Gly-Thr-DPhe-Arg-Wpb-Pen]-Abu

[0464] SEQ ID NO: 100 (e.g., CTX-1796) - Arg-c[Cys-Gly-Thr-DPhe-Arg-Wpb-Pen]

[0465] SEQ ID NO: 101 (e.g., CTX-1797) - Arg-c[Pen-Gly-Cit-DPhe-Arg-Wpb-Cys]-Abu

[0466] SEQ ID NO: 102 (e.g., CTX-1798) - Arg-c[Pen-Gly-Cit-DPhe-Arg-Wpb-Cys]

[0467] SEQ ID NO: 103 (e.g., CTX-1799) - Arg-c[Pen-Gly-Thr-DPhe-Arg-Wpb-Pen]-Abu

[0468] SEQ ID NO: 104 (e.g., CTX-1800) - Arg-c[Pen-Gly-Thr-DPhe-Arg-Wpb-Pen]

[0469] SEQ ID NO: 105 (e.g., CTX-1801) - Arg-c[hCys-Gly-Cit-DPhe-Arg-Wpb-Pen]-Abu

[0470] SEQ ID NO: 106 (e.g., CTX-1802) - Arg-c[hCys-Gly-Cit-DPhe-Arg-Wpb-Pen]

[0471] SEQ ID NO: 107 (e.g., CTX-1803) - Arg-c[hCys-Gly-Thr-DPhe-Arg-Wpb-Pen]-Abu

[0472] SEQ ID NO: 108 (e.g., CTX-1804) - Arg-c[hCys-Gly-Thr-DPhe-Arg-Wpb-Pen]

[0473] SEQ ID NO: 109 (e.g., CTX-1805) - c[hCys-Aib-Thr-DPhe-Arg-Wpb-Pen]

[0474] SEQ ID NO: 110 (e.g., CTX-1806) - c[hCys-Gly-Cit-DPhe-Arg-Wpb-Pen]

[0475] SEQ ID NO: 111 (e.g., CTX-1807) - c[Cys-Aib-Cit-DPhe-Arg-Wpb-Cys]

[0476] REFERENCES

[0477] The following references are herein incorporated by reference in their entireties.

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[0517] 33. Carotenuto, A. et al. Discovery of Novel Potent and Selective Agonists at the Melanocortin-3 Receptor. J. Med. Chem. (2015). doi:10.1021 / acs.jmedchem.5b01285 34. Herpertz-dahlmann, B., Holtkamp, K. & Konrad, K. Eating disorders: Anorexia and bulimia nervosa. Handb. Clin. Neurol. (2012). doi:10.1016 / B978-0-444-52002- 9.00026-7

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[0519] 36. Jerlhag, E. et al. Ghrelin stimulates locomotor activity and accumbal dopamine- overflow via central cholinergic systems in mice: Implications for its involvement in brain reward. Addict. Biol. (2006). doi:10.1111 / j.l369-1600.2006.00002.x

[0520] 37. Adermark, L. et al. Ghrelin administration into tegmental areas stimulates locomotor activity and increases extracellular concentration of dopamine in the nucleus accumbens. Addict. Biol. (2007). doi:10.1111 / j.l369-1600.2006.00041.x

[0521] 38. Marks, D. L., Hruby, V., Brookhart, G. & Cone, R. D. The regulation of food intake by selective stimulation of the type 3 melanocortin receptor (MC3R). Peptides (2006). doi: 10.1016 / j .peptides.2005.01.025

[0522] 39. Lee, M. et al. Effects of selective modulation of the central melanocortin-3 -receptor on food intake and hypothalamic POMC expression. Peptides (2008).

[0523] doi: 10.1016 / j .peptides.2007.11.005

[0524] 40. Lippert, R. N., Ellacot, K. L. J. & Cone, R. D. Gender-specific roles for the melanocortin-3 receptor in the regulation of the mesolimbic dopamine system in mice. Endocrinology (2014). doi:10.1210 / en.2013-2049

[0525] 41. Pandit, R. et al. Melanocortin 3 receptor signaling in midbrain dopamine neurons increases the motivation for food reward. Neuropsychopharmacology (2016).

[0526] doi:10.1038 / npp.2016.19Atorney Docket No. UM-43572.601

[0527] Client Ref. No. UM 2025-153 Mavrikaki, M. et al. Melanocortin-3 receptors in the limbic system mediate feeding-related motivational responses during weight loss. Mol. Metab. (2016). doi:10.1016 / j.molmet.2016.05.002

[0528] Pei, H. et al. Lateral Hypothalamic Mc3R-Expressing Neurons Modulate Locomotor Activity, Energy Expenditure, and Adiposity in Male Mice. Endocrinology 160, 343-358 (2018).

[0529] Stemson, S. M. & Eiselt, A.-K. Three Pillars for the Neural Control of Appetite. Annu. Rev. Physiol. (2016). doi:10.1146 / annurev-physiol-021115-104948

[0530] Rossi, M. A. & Stuber, G. D. Overlapping Brain Circuits for Homeostatic and Hedonic Feeding. Cell Metabolism (2018). doi:10.1016 / j.cmet.2017.09.021

[0531] Sweeney, P. & Yang, Y. Neural Circuit Mechanisms Underlying Emotional Regulation of Homeostatic Feeding. Trends in Endocrinology and Metabolism (2017). doi:10.1016 / j.tem.2017.02.006

[0532] Hay, P. J., Touyz, S. & Sud, R. Treatment for severe and enduring anorexia nervosa: A review. Australian and New Zealand Journal of Psychiatry (2012).

[0533] doi: 10.1177 / 0004867412450469

[0534] Bulik, C. M. et al. Prevalence, heritability, and prospective risk factors for anorexia nervosa. Arch. Gen. Psychiatry (2006). doi:10.1001 / archpsyc.63.3.305

[0535] Keski-Rahkonen, A. et al. Epidemiology and course of anorexia nervosa in the community. Am. J. Psychiatry (2007). doi:10.1176 / appi.ajp.2007.06081388 Marks, D. L., Butler, A. A., Turner, R., Brookhart, G. & Cone, R. D. Differential role of melanocortin receptor subtypes in cachexia. Endocrinology (2003).

[0536] doi: 10.1210 / en.2002-221099

[0537] Fazeli, P. K. et al. Treatment with a ghrelin agonist in outpatient women with anorexia nervosa: A randomized clinical trial. J. Clin. Psychiatry (2018).

[0538] doi: 10.4088 / JCP.17ml 1585

[0539] Fazeli, P. et al. Short-term treatment with a ghrelin agonist significantly improves gastric emptying in anorexia nervosa. Endocrine reviews. Conference: 98th annual meeting and expo of the endocrine society, ENDO 2016. United states. Conference start: 20160401. Conference end: 20160404 (2016). doi:10.1210 / endo-meetings.2016.OABA.2. SUN-606

[0540] Carlini, V. P. et al. Ghrelin increases anxiety-like behavior and memory retention in rats. Biochem. Biophys. Res. Commun. (2002).

[0541] Carvajal, P., Carlini, V. P., Schioth, H. B., de Barioglio, S. R. & Salvatierra, N. A. Central ghrelin increases anxiety in the Open Field test and impairs retention memory in a passive avoidance task in neonatal chicks. Neurobiol. Learn. Mem. (2009).Atorney Docket No. UM-43572.601

[0542] Client Ref. No. UM 2025-153 doi:10.1016 / j.nlm.2008.12.008

[0543] Sweeney, P. & Yang, Y. An excitatory ventral hippocampus to lateral septum circuit that suppresses feeding. Nat. Commun. (2015). doi:10.1038 / ncommsl0188 Sweeney, P. & Yang, Y. An Inhibitory Septum to Lateral Hypothalamus Circuit That Suppresses Feeding. J. Neurosci. (2016). doi:10.1523 / jneurosci.2042-16.2016 Sweeney, P., Li, C. & Yang, Y. Appetite suppressive role of medial septal glutamatergic neurons. Proc. Natl. Acad. Sci. (2017). doi:10.1073 / pnas.1707228114 Singh, A., Dirain, M., Witek, R., Rocca J.R., Edison, A.S., and Haskell-Luevano, C. Structure- Activity Relationships of Peptides Incorporating a Bioactive Reverse-Turn Heterocycle at the Melanocortin Receptors: Identification of a 5800-fold Mouse Melanocortin-3 Receptor (mMC3R) Selective Antagonist / Partial Agonist versus the Mouse Melanocortin-4 Receptor (mMC4R). J. Med. Chem. 56, 2747-2763, 2013.

Claims

Atorney Docket No. UM-43572.601Client Ref. No. UM 2025-153 CLAIMS1. A composition comprising a peptide having 4 or fewer substitutions relative to SEQ ID NO: 1;wherein AA1 is linked to AA2 if AA1B is absent;wherein the peptide comprises at least one of:bAla at AAO;Phe, Hph, Gly, Ala at AA1B;Pro, D-Nle, D-Hle, D-Alb, or beta- Ala at AA3;Agp, Nar, Cit, or Alb at AA4;D-Fpy, D-Fpt, or D-Fpm at AA5;Wpb at AA7; orAbu, Hie, Cpa, Phg, D-Hph, or Om at AA9.

2. The composition of claim 1, wherein the peptide comprises 100% sequence similarity to SEQ ID NO: 1.

3. The composition of claim 1, wherein the peptide comprises SEQ ID NO: 1.

4. A composition comprising a peptide having 4 or fewer substitutions relative to SEQ ID NO: 2;wherein AA1 is linked to AA2 if AA1B is absent;wherein the peptide comprises at least one of:bAla at AAO;Phe, Hph, Gly, Ala at AA1B;Pro, D-Nle, D-Hle, D-Alb, or beta- Ala at AA3;Agp, Nar, Cit, or Alb at AA4;D-Fpy, D-Fpt, or D-Fpm at AA5;Wpb at AA7; orAbu, Hie, Cpa, Phg, D-Hph, or Om at AA9.

5. The composition of claim 4, wherein the peptide comprises 100% sequence similarity to SEQ ID NO: 2.Atorney Docket No. UM-43572.601Client Ref. No. UM 2025-153 6. The composition of claim 4, wherein the peptide comprises SEQ ID NO: 2.

7. The composition of one of claims 1-6, wherein AA4 is Alb.

8. The composition of claim 1-6, wherein AA4 is Cit.

9. The composition of one of claims 1-8, wherein AA7 is Wpb.

10. The composition of claim 1, wherein the peptide has 4 or fewer substitutions relative to one of SEQ ID NOS: 3-22, 42-71, 81-86, and / or 95-111.

11. The composition of claim 1, wherein the peptide has 100% sequence similarity with one of SEQ ID NOS: 3-22, 42-71, 81-86, and / or 95-111.

12. The composition of claim 11, wherein the peptide comprises the sequence of one of SEQ ID NOS: 3-22, 42-71, 81-86, or 95-111.

13. A composition comprising a peptide having 4 or fewer substitutions relative to SEQ ID NO: 90;wherein the peptide comprises at least one of:Aib, D-Nle, D-Alb, or beta- Ala at AA3;Agp, Nar, Cit, or Alb at AA4;D-Fpy at AA5; orWpb at AA7.

14. The composition of claim 13, wherein the peptide comprises 100% sequence similarity to SEQ ID NO: 90.

15. The composition of claim 13, wherein the peptide comprises SEQ ID NO: 90.

16. A composition comprising a peptide having 4 or fewer substitutions relative to SEQ ID NO: 91;wherein the peptide comprises at least one of:Aib, D-Nle, D-Alb, or beta- Ala at AA3;Atorney Docket No. UM-43572.601Client Ref. No. UM 2025-153 Agp, Cit, or Nar, at AA4; orWpb at AA7.

17. The composition of claim 16, wherein the peptide comprises 100% sequence similarity to SEQ ID NO: 91.

18. The composition of claim 16, wherein the peptide comprises SEQ ID NO: 91.

19. The composition of one of claims 13-18, wherein AA7 is Wpb.

20. The composition of claim 13, wherein the peptide has 4 or fewer substitutions relative to one of SEQ ID NOS: 23-41, 72-80, 87-89 and / or 93-94.

21. The composition of claim 13, wherein the peptide has sequence similarity with one of SEQ ID NOS: 23-41, 72-80, 87-89 and / or 93-94.

22. The composition of claim 21, wherein the peptide comprises the sequence of one of SEQ ID NOS: 23-41, 72-80, 87-89 or 93-94.

23. The composition of one of claims 1-22, wherein the peptide a melanocortin 4 receptor (MC4R) agonist.

24. The composition of one of claims 1-23, wherein the composition is orally bioavailable.

25. A pharmaceutical formulation comprising the composition of one of claims 1-24 and a pharmaceutically acceptable carrier.

26. The pharmaceutical composition of claim 25, wherein the composition is present at a subtherapeutic dose.

27. The pharmaceutical composition of claim 25, wherein the composition is present at a substandard dose.Attorney Docket No. UM-43572.601Client Ref. No. UM 2025-153 28. The pharmaceutical composition of claim 25, farther comprising an antiobesity agent.

29. The pharmaceutical composition of claim 28, wherein the anti-obesity agent is selected from a GLP-1 agonist, an anorectic, a pro-satiety agent, a lipase inhibitor, and combinations thereof.

30. The pharmaceutical composition of claim 29, wherein the anti-obesity agent is a GLP-1 agonist.

31. The pharmaceutical composition of claim 30, wherein the GLP-1 agonist is selected from exendin-4, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, semaglutide, taspoglutide, tirzepatide, retatrutide, CNTO736, CNT03649, HM11260C (LAPS-Exendin), NN9926 (OG9S7GT), TT401, ZYOG1, maritide, orforglipron, NN-9932, andHRS-953.

32. The pharmaceutical composition of claim 29, wherein the anti-obesity agent is an anorectic.

33. The pharmaceutical composition of claim 32, wherein the anorectic is selected from amphetamine, dexamphetamine, amfepramone, clobenzorex, mazindol, phentermine, phentermine and topiramate, and lorcaserin.

34. The pharmaceutical composition of claim 29, wherein the anti-obesity agent is a pro-satiety agent.

35. The pharmaceutical composition of claim 34, wherein the pro-satiety agent is selected from neurotrophic factor, amylin, calcitonin, cholecystokinin (CCK), leptin, metreleptin oxyntomodulin, pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), pramlitide, cagrilintide, eloralintide, petrelintide, AZD6234, GUB014295, and KBP336.

36. The pharmaceutical composition of claim 29, wherein the anti-obesity agent is a lipase inhibitor.Attorney Docket No. UM-43572.601Client Ref. No. UM 2025-15337. The pharmaceutical composition of claim 36, wherein the lipase inhibitor is selected from caulerpenyne, cetilistat, ebelactone A and B, esterastin, lipstatin, orlistat, percyquinin, panclicin A-E, valilactone and vibralactone.

38. The pharmaceutical composition of claim 29, wherein the anti-obesity agent is a combination of two or more agents selected from phentermine / topiramate, naltrexone / bupropion, cagrilintide / semaglutide, amycretin, retatrutide, survodutide, pemvidutide, and mazdutide.

39. The pharmaceutical composition of one of claims 28-38, wherein the antiobesity agent is present at a sub-therapeutic dose.

40. The pharmaceutical composition of one of claims 28-38, wherein the antiobesity agent is present at a sub-standard dose.

41. The pharmaceutical composition of one of claims 28-40, wherein the peptide is present at a sub-therapeutic dose.

42. The pharmaceutical composition of one of claims 28-40, wherein the peptide is present at a sub-standard dose.

43. A method of treating a subject for a disease, condition, or disorder comprising administering a composition of one of claims 1-24 or a pharmaceutical composition of one of claims 25-42 to a subject.

44. The method of claim 43, wherein the subject suffers from positive energy balance as the cause or result of the disease, condition, or disorder.

45. The method of claim 43, wherein the disease, condition, or disorder is characterized by overeating.Atorney Docket No. UM-43572.601Client Ref. No. UM 2025-153 46. The method of claim 43, wherein the disease, condition, or disorder characterized by one or more emotional / mental symptoms.

47. The method of claim 43, wherein the disease, condition, or disorder is caused by or is the result of obesity.

48. The method of claim 43, wherein the subject suffers from diabetes, heart disease, hypertension, sleep apnea, depression, kidney disease, and / or arthritis.

49. The method of claim 43, wherein the subject suffers from genetic obesity.

50. The method of claim 49, wherein the subject suffers from congenital leptin deficiency, leptin receptor deficiency, MC4R deficiency, POMC deficiency, PCSK1 deficiency, LEPR deficiency, Chung-Jansen syndrome, Ulnar-Mammary syndrome, PLXNA deficiency, SRC1 deficiency, heterozygous POMC / PSK1 deficiency, hypothalamic obesity, heterozygous LEPR deficiency, MC4R haploiud insufficiency, Prader-Willi syndrome, Bardet-Biedl syndrome, Alstrom syndrome, Cohen syndrome, Smith-Magenis syndrome, 16pl 1.2 deletion syndrome, ASIP obesity syndrome, Kleine-Levin, binge eating disorders, Down syndrome, Albright’s hereditary osteodystrophy with pseudohypoparathy, loss of fimction ADCY3, SIM1 deficiency, lp36 deletion distal syndrome, or WAGR syndrome.

51. The method of one of claims 43-50, wherein the composition is coadministered with nutritional therapy, psychotherapy, or other pharmaceutical agents.

52. The method of claim 51 , wherein the composition is co-administered with an anti-obesity agent.

53. The method of claim 52, wherein the anti-obesity agent is selected from a GLP-1 agonist, an anorectic, a pro-satiety agent, a lipase inhibitor, and combinations thereof.

54. The method of claim 53, wherein the anti-obesity agent is a GLP-1 agonist.

55. The method of claim 54, wherein the GLP-1 agonist is selected from exendin-4, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, semaglutide, taspoglutide,Atorney Docket No. UM-43572.601Client Ref. No. UM 2025-153 tirzepatide, retatrutide, CNTO736, CNT03649, HM11260C (LAPS-Exendin), NN9926 (OG9S7GT), TT401, ZYOG1, maritide, orforglipron, NN-9932, and HRS-953.

56. The method of claim 53, wherein the anti-obesity agent is an anorectic.

57. The method of claim 56, wherein the anorectic is selected from amphetamine, dexamphetamine, amfepramone, clobenzorex, mazindol, phentermine, phentermine and topiramate, and lorcaserin.

58. The method of claim 53, wherein the anti-obesity agent is a pro-satiety agent.

59. The method of claim 58, wherein the pro-satiety agent is selected from neurotrophic factor, amylin, calcitonin, cholecystokinin (CCK), leptin, metreleptin oxyntomodulin, pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), pramlitide, cagrilintide, eloralintide, petrelintide, AZD6234, GUB014295, and KBP336.

60. The method of claim 53, wherein the anti-obesity agent is a lipase inhibitor.

61. The method of claim 60, wherein the lipase inhibitor is selected from caulerpenyne, cetilistat, ebelactone A and B, esterastin, lipstatin, orlistat, percyquinin, panclicin A-E, valilactone and vibralactone.

62. The method of claim 53, wherein the anti-obesity agent is a combination of two or more agents selected from phentermine / topiramate, naltrexone / bupropion, cagrilintide / semaglutide, amycretin, retatrutide, survodutide, pemvidutide, and mazdutide.

63. The method of claim one of claims 52-62, wherein the anti-obesity agent is administered at a sub-therapeutic dose.

64. The method of claim one of claims 52-62, wherein the anti-obesity agent is administered at a sub-standard dose.

65. The method of claim one of claims 52-64, wherein the peptide is administered at a sub-therapeutic dose.Atorney Docket No. UM-43572.601Client Ref. No. UM 2025-15366. The method of claim one of claims 52-64, wherein the peptide is administered at a sub-standard dose.

67. The method of one of claims 43-66, wherein the administration is repeated on a recurring basis for a period of at least 1 week.

68. The method of claim 67, wherein the administration is repeated on a daily basis.

69. The method of claim 67, wherein the administration is repeated on a recurring basis for a period of at least 1 month.

70. The method of claim 69, wherein the administration is repeated on a recurring basis for a period of at least 1 year.

71. The method of one of claims 43-70, wherein the method results in a reduction of mass to the subject.

72. The method of claim 71, wherein the greater than 50% of the reduction in mass is loss of fat.

73. The method of claim 72, wherein the greater than 75% of the reduction in mass is loss of fat.

74. The method of one of claims 43-73, wherein at least 25% of the reduction of mass is maintained 3 months after administering the composition to the subject.

75. Use of a composition of one of claims 1-24 or a pharmaceutical composition of one of claims 25-42 in the treatment or prevention of a condition, disease, or disorder.

76. Use of a composition of one of claims 1-24 or a pharmaceutical composition of one of claims 25-42 as a medicament.Atorney Docket No. UM-43572.601Client Ref. No. UM 2025-153 77. Use of a composition of one of claims 1-24 or a pharmaceutical composition of one of claims 25-42 for use in the manufacture of a medicament.

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