Long-acting amylin receptor agonists and uses thereof

Abstract

The present disclosure relates to the field of medicine. In particular, the present disclosure belongs to the field of diabetes, obesity and / or chronic weight management, dyslipidemia, and / or NASH treatment. The present disclosure relates to compounds that can stimulate amylin receptors to reduce food intake, body weight, blood glucose, and / or triglycerides, and thus can be used to treat diabetes, obesity, and / or dyslipidemia. The present disclosure also includes pharmaceutical compositions containing such compounds, and the therapeutic use of such compounds and compositions.

Description

[Technical field]

[0001] The present disclosure relates to the field of medicine. In particular, the present disclosure is in the field of treating diabetes, obesity and / or chronic weight control, dyslipidemia and / or non-alcoholic steatohepatitis (NASH). The present disclosure relates to compounds that stimulate amylin receptors and thus can reduce food intake, body weight, glucose HbA1c (glycated hemoglobin), and / or triglycerides, and can be used to treat diabetes, obesity and / or chronic weight control, dyslipidemia, and / or NASH. The present disclosure also includes pharmaceutical compositions containing such compounds, and therapeutic uses of such compounds and pharmaceutical compositions. [Background technology]

[0002] Over the past few decades, the prevalence of diabetes has been increasing. Type 2 diabetes mellitus ("T2DM") is the most common form of diabetes, accounting for 90% of all diabetes cases. T2DM is characterized by high blood glucose levels and is primarily associated with insulin resistance. The desired treatment for diabetic patients should lower blood glucose levels and stabilize HbA1c.

[0003] Obesity is a complex medical disorder that leads to excessive accumulation of adipose tissue mass.Today, obesity is a global public health concern that is often associated with undesirable health outcomes and morbidity.In the desired treatment of obese patients, excess weight should be reduced, obesity-related comorbidities should be improved, and / or long-term weight loss should be maintained.

[0004] Dyslipidemia is abnormal levels of cholesterol and other lipids (also called fats) in the blood. Dyslipidemia increases the chances of developing clogged arteries (atherosclerosis) and heart attacks, strokes, or other cardiovascular problems, especially in individuals who smoke.

[0005] NASH stands for non-alcoholic steatohepatitis. It is a hepatic manifestation of metabolic disorders and is the most severe form of non-alcoholic fatty liver disease (NAFLD). NASH is closely related to the comorbidities of diabetes and obesity.

[0006] Amylin is a 37 amino acid peptide hormone that is co-secreted with insulin from the beta cells of the pancreas and is deficient in people with diabetes. It inhibits glucagon secretion, slows gastric emptying, and acts as a satiety agent. As a result, amylin helps regulate the amount of glucose in the body after a meal. However, the physiochemical properties of human amylin make its use as a medicine difficult. For example, amylin is difficult to formulate because it is chemically unstable and precipitates at physiological pH, necessitating formulation in acidic solutions. Furthermore, the half-life of the drug is less than one hour, and because the drug is used at mealtimes, patients would need to take the drug multiple times a day to use it as a therapy.

[0007] Pramlintide is a commercially available amylin agonist peptide used in the treatment of diabetes as an add-on to insulin. Pramlintide is chemically unstable at neutral pH due to both the presence of disulfide bonds (-SS-) and deamidation. It is therefore provided in an acidic formulation. Compared to human amylin, the amino acids at positions 25, 28 and 29 of pramlintide are replaced with prolines. These modifications reduce the tendency of the peptide towards fibril formation. However, pramlintide still has a very short plasma half-life and therefore must be injected 2-3 times a day, which is inconvenient and therefore leads to poor adherence to dosing frequency, which may further result in incomplete or suboptimal efficacy of the drug. In addition, its low pH formulation is not compatible with the neutral pH formulations used for insulin and GLP-1 analogues, thereby complicating its co-administration with such compounds, which may themselves be synergistic and therefore have improved clinical efficacy.

[0008] Human amylin binds to two different receptor complexes. These two complexes also contain calcitonin receptor and receptor activating protein, RAMP1 or RAMP3. Calcitonin receptors are found in many tissues throughout the body and are thought to be involved in the regulation of bone metabolism. However, little is known about the physiological function of calcitonin receptors in humans other than bone regulation, and therefore, molecules with high affinity for calcitonin receptors may increase the risk of off-target toxicity. Therefore, it is believed that amylin-based polypeptides with increased selectivity for amylin receptors compared to calcitonin receptors may provide advantageous pharmacokinetic and pharmacological profiles.

[0009] Due to the close relationship between calcitonin and amylin receptors, some cross-reactivity to calcitonin receptors may be expected for amylin receptor agonists. As an example, the amylin agonist peptide pramlintide is 14-fold more potent than amylin receptors, but has some affinity for calcitonin receptors.

[0010] Polypeptides comprising human amylin agonist peptides and having an albumin binding moiety have been disclosed. See WO 2010 / 046357, WO 2009 / 034119, and WO 2009 / 034119. Although these polypeptides with albumin binding moieties show improved pharmacokinetic (PK) or pharmacodynamic (PD) properties compared to pramlintide, they may still show poor physical stability under certain conditions. Furthermore, polypeptides generally do not show selectivity for amylin receptors over calcitonin receptors. US 2014 / 0087995 relates to polypeptides comprising amylin agonist peptides having selectivity for amylin receptors over calcitonin receptors. However, there remains a need for amylin agonist peptide drug candidates with higher potency, lower developability risk, lower immunogenicity risk, improved chemical stability, and compatibility with formulations at neutral pH.

[0011] There is a need for compounds (e.g., peptides) that exhibit selectivity for agonism of amylin receptors over calcitonin receptors. Additionally, there is a need for amylin receptor agonists with extended duration of action and retained efficacy. Therapeutically desirable compounds will stimulate amylin receptors and provide one or more advantageous properties, such as reduced food intake, reduced body weight, reduced blood glucose levels, reduced HbA1c, reduced triglycerides, and / or reduced insulin levels. Additionally, therapeutically desirable compounds may have one or more additional advantageous properties, such as a long duration of action with retained or improved efficacy in stimulating amylin receptors, a low risk of immunogenic response, and / or a low risk of fibrillation.

[0012] Furthermore, a combination of the amylin receptor agonist of the present disclosure, optionally in combination with an incretin or incretin analog, is desirable to provide treatment for diabetes, obesity, NASH, and / or dyslipidemia. Such a combination will also preferably be more effective than either molecule alone. For example, such treatment with such a combination may allow for the use of lower doses of either or both molecules compared to each molecule alone, potentially resulting in fewer side effects (or shorter duration of one or the other therapy) while maintaining efficacy. It is believed that the novel combination(s) provided herein will be an effective treatment for diabetes, obesity, NASH, and / or dyslipidemia. Summary of the Invention

[0013] Accordingly, the present disclosure provides a method for treating diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of an amylin receptor agonist of the present invention and an effective amount of an additional agent.

[0014] Thus, the present disclosure describes novel compounds (peptides) that stimulate amylin receptors and have one or more of the following properties: (1) effective reduction in food intake and body weight; (2) lowering of glucose and insulin; (3) reduced immunogenicity risk and low risk of fibrillation compared to pramlintide; (4) significantly extended half-life compared to pramlintide. The extended half-life of the compounds disclosed herein allows these compounds to be dosed only once a week when used in therapy. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] One embodiment of the present disclosure comprises: Xaa 1 -C-Xaa 3 -TATCAT-Xaa 10 -Xaa 11 -Xaa 12 -AE-Xaa15 -LVRSS-Xaa 21 -Xaa 22 -FGP-Xaa 26 -LPPTEVGSNTY-NH 2 (In the sequence, Xaa 1 is K or γE, and Xaa 3 is E, N, or G; Xaa 10 is G or Q, and Xaa 11 is Orn or K, and Xaa 12 is L or αMeL, and Xaa 15 is αMeF or F, and Xaa 21 is N or H, and Xaa 22 is NMeD, NMeN, or N; Xaa 26 is I or K) (SEQ ID NO: 14) or a pharma- ceutically acceptable salt thereof. Optionally, the compound comprising SEQ ID NO: 14 further comprises additional elements to extend the time-action profile of the compound. These additional elements include an Fc portion of an immunoglobulin, a fragment of an Fc portion of an immunoglobulin, human serum albumin (HSA), a variant of a VHH (variable domain of a heavy chain antibody), a nanobody, a fragment of human serum albumin, a C, or a pharma- ceutically acceptable salt thereof, with or without a linker, at any suitable position in the sequence. 20 Monoacid, C 20 The additional moieties include diacids, and polyethylene glycol (PEG) moieties, or other types of high molecular weight polymers. Preferably, the additional moieties are attached to lysines on the compounds, optionally using linkers. Furthermore, in any of the embodiments disclosed herein, the amylin agonist peptides and compounds containing amylin agonist peptides may further include such moieties, either in addition to or as a replacement for any already present moieties that are attached or linked to the peptide sequence and extend the time-action profile.

[0016] In some embodiments of the compound comprising SEQ ID NO: 14, Xaa 26 is lysine. In some embodiments of compounds comprising SEQ ID NO: 14, the lysine at position 26, if present, is of the formula: (γE)2 -CO-(CH 2 ) 18 -CO 2 H. In some embodiments of compounds comprising SEQ ID NO: 14, the lysine at position 26, if present, is represented by the formula: AEEA 2 -γE-CO-(CH 2 ) 18 -CO 2 H. In some embodiments of compounds comprising SEQ ID NO: 14, the lysine at position 26, if present, is represented by the formula: γE-AEEA 2 -CO-(CH 2 ) 18 -CO 2 H. In some embodiments of compounds comprising SEQ ID NO: 14, the lysine at position 26, if present, is represented by the formula: (γE) 2 -AEEA-CO-(CH 2 ) 18 -CO 2 The amylin agonist peptide is attached to the fatty acid via a linker shown in formula H. In other embodiments of the compound comprising SEQ ID NO: 14, the lysine at position 26, if present, is attached to the fatty acid via another linker known in the art. Additionally, in any of the embodiments disclosed herein, the amylin agonist peptides and compounds comprising amylin agonist peptides may include another linker for attaching the fatty acid as shown in the formulas herein.

[0017] In some embodiments of the compound comprising SEQ ID NO: 14, Xaa 1 is lysine. In some embodiments of compounds comprising SEQ ID NO: 14, the lysine at position 1, if present, is of the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2H. In other embodiments of compounds comprising SEQ ID NO: 14, the lysine at position 1, if present, is attached to the fatty acid via another linker known in the art.

[0018] In some embodiments of the compound comprising SEQ ID NO: 14, there is a disulfide bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7. In preferred embodiments of the compound comprising SEQ ID NO: 14, there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7.

[0019] One embodiment herein includes the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeN-FGPKLPPTEVGSNTY-NH 2 (SEQ ID NO:1). An alternative embodiment herein consists of the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeN-FGPKLPPTEVGSNTY-NH 2 (SEQ ID NO:1). Below is a depiction of compound I using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (where the peptide bond is formed with the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeN at position 22, and tyrosine at position 37:

[0020] [ka] Compound I; SEQ ID NO:1

[0021] Alternatively, the compound can have at least about 90% to about 99% sequence similarity to SEQ ID NO: 1. Alternatively, the compound can have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 1.

[0022] In some embodiments of the compound comprising SEQ ID NO:1, the lysine at position 26 is linked to a fatty acid via a linker. In a preferred embodiment of the compound comprising SEQ ID NO:1, the lysine at position 26 is represented by the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 It is attached to the fatty acid via a linker, represented by H.

[0023] One embodiment herein includes the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeN-FGPKLPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is represented by the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO: 2) to the fatty acid. An alternative embodiment herein consists of the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeN-FGPKLPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is represented by the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:2) to the fatty acid. Below is a depiction of compound II using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (the peptide bond is formed using the side chain carboxylic acid at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeN at position 22, lysine at position 26, and tyrosine at position 37:

[0024] [ka] Compound II; SEQ ID NO:2

[0025] Alternatively, the compound can have at least about 90% to about 99% sequence similarity to SEQ ID NO: 2. Alternatively, the compound can have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 2.

[0026] One embodiment herein includes the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPKLPPTEVGSNTY-NH 2 (SEQ ID NO:3). An alternative embodiment herein consists of the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPKLPPTEVGSNTY-NH 2 (SEQ ID NO:3). Below is a depiction of compound III using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (where the peptide bond is formed with the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, and tyrosine at position 37:

[0027] [ka] Compound III; SEQ ID NO:3

[0028] Alternatively, the compound can have at least about 90% to about 99% sequence similarity to SEQ ID NO: 3. Alternatively, the compound can have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to compound III (SEQ ID NO: 3).

[0029] In some embodiments of the compound comprising SEQ ID NO: 3, the lysine at position 26 is linked to a fatty acid via a linker. In a preferred embodiment of the compound comprising SEQ ID NO: 3, the lysine at position 26 is represented by the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H is attached to the fatty acid linker moiety.

[0030] One embodiment herein includes the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPKLPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is represented by the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO: 4). An alternative embodiment herein is a linker moiety having the following sequence: γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPKLPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is of the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:4) is attached to a fatty acid linker moiety. Below is a depiction of compound IV using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, lysine at position 26, and tyrosine at position 37:

[0031] [ka] Compound IV; SEQ ID NO:4

[0032] Alternatively, the compound can have at least about 90% to about 99% sequence similarity to SEQ ID NO: 4. Alternatively, the compound can have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 4.

[0033] One embodiment herein includes the sequence: KCETATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPILPPTEVGSNTY-NH 2 (SEQ ID NO: 5). An alternative embodiment herein is the following sequence: KCETATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPILPPTEVGSNTY-NH 2 (SEQ ID NO:5). Below is a depiction of compound V using standard single letter amino acid code, except that the structures of the amino acid residues at cysteine ​​positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, and tyrosine at position 37 have been expanded:

[0034] [ka] Compound V; SEQ ID NO:5

[0035] Alternatively, the compound may have at least about 90% to about 99% sequence similarity to SEQ ID NO: 5. Alternatively, the compound may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 5.

[0036] In some embodiments of the compound comprising SEQ ID NO:5, the lysine at position 1 is linked to a fatty acid via a linker. In a preferred embodiment of the compound comprising SEQ ID NO:5, the lysine at position 1 is represented by the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2It is attached to the fatty acid via a linker, represented by H.

[0037] One embodiment herein is the following sequence: KCETATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPILPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 1 is of the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO: 6). An alternative embodiment herein is a nucleotide sequence of the following sequence: KCETATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPILPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 1 is of the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:6). Below is a depiction of compound VI using standard single letter amino acid code, except that the structures of the amino acid residues at lysine at position 1, cysteine ​​at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, and tyrosine at position 37 have been expanded:

[0038] [ka] Compound VI: SEQ ID NO:6

[0039] Alternatively, the compound can have at least about 90% to about 99% sequence similarity to SEQ ID NO: 6. Alternatively, the compound can have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to compound VI (SEQ ID NO: 6).

[0040] One embodiment herein includes the sequence: KCETATCATG-Orn-αMeL-AEFLVRSSHNFGPILPPTEVGSNTY-NH 2 (SEQ ID NO: 7). An alternative embodiment herein is the following sequence: KCETATCATG-Orn-αMeL-AEFLVRSSHNFGPILPPTEVGSNTY-NH 2 (SEQ ID NO: 7). The following is a depiction of compound VII, except that the structures of the amino acid residues Cysteine ​​at positions 2 and 7, Orn at position 11, αMeL at position 12, and Tyrosine at position 37 are expanded:

[0041] [ka] Compound VII; SEQ ID NO:7

[0042] Alternatively, the compound may have at least about 90% to about 99% sequence similarity to SEQ ID NO: 7. Alternatively, the compound may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 7.

[0043] In some embodiments of the compound comprising SEQ ID NO: 7, the lysine at position 1 is linked to a fatty acid linker moiety. In a preferred embodiment of the compound comprising SEQ ID NO: 7, the lysine at position 1 is linked to a fatty acid linker moiety of the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 It is attached to the fatty acid via a linker, represented by H.

[0044] One embodiment herein is the following sequence: KCETATCATG-Orn-αMeL-AEFLVRSSHNFGPILPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 1 is of the formula: (γE) 2 -CO-(CH 2 ) 18 -CO2 H (SEQ ID NO: 8). An alternative embodiment herein is a linker having the following sequence: KCETATCATG-Orn-αMeL-AEFLVRSSHNFGPILPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 1 is of the formula: (γE) 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:8) to the fatty acid. Below is a depiction of compound VIII using the standard single letter amino acid code, except that the structures of the amino acid residues Lysine at position 1, Orn at positions 2 and 7, αMeL at position 12, and Tyrosine at position 37 have been expanded:

[0045] [ka] Compound VIII; SEQ ID NO:8

[0046] Alternatively, the compound may have at least about 90% to about 99% sequence similarity to SEQ ID NO: 8. Alternatively, the compound may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 8.

[0047] One embodiment herein is the following sequence: γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is of the formula: 2 -γE-CO-(CH 2 ) 18 -CO 2H (SEQ ID NO: 9). An alternative embodiment herein is a linker having the following sequence: γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is of the formula: 2 -γE-CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:9) is attached to the fatty acid via a linker. Below is a depiction of compound IX using the standard single letter amino acid code, except that the structures of the amino acid residues are expanded for glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, lysine at position 26, and tyrosine at position 37:

[0048] [ka] Compound IX; SEQ ID NO:9

[0049] Alternatively, the compound may have at least about 90% to about 99% sequence similarity to SEQ ID NO: 9. Alternatively, the compound may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 9.

[0050] One embodiment herein includes the following sequence: γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is represented by the formula: γE-AEEA 2 -CO-(CH 2 ) 18 -CO 2H (SEQ ID NO: 10). An alternative embodiment herein is a linker having the following sequence: γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is of the formula: γE-AEEA 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:10). Below is a depiction of compound X using the standard single letter amino acid code, except that for glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, lysine at position 26, and tyrosine at position 37, the structures of those amino acid residues have been expanded:

[0051] [ka] Compound X; SEQ ID NO: 10

[0052] Alternatively, the compound can have at least about 90% to about 99% sequence similarity to SEQ ID NO: 10. Alternatively, the compound can have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 10.

[0053] One embodiment herein includes the following sequence: γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is represented by the formula: (γE) 2 -AEEA-CO-(CH 2 ) 18 -CO 2H (SEQ ID NO: 11). An alternative embodiment herein is a γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is of the formula: (γE) 2 -AEEA-CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:11). Below is a depiction of compound XI using the standard single letter amino acid code, except that for glutamic acid (γE) at position 1 (wherein the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, lysine at position 26, and tyrosine at position 37, the structures of those amino acid residues have been expanded:

[0054] [ka] Compound XI; SEQ ID NO:11

[0055] Alternatively, the compound can have at least about 90% to about 99% sequence similarity to SEQ ID NO: 11. Alternatively, the compound can have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 11.

[0056] One embodiment herein includes the following sequence: γE-CNTATCATQ-Orn-LAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is represented by the formula: 2 -γE-CO-(CH 2 ) 18 -CO 2H (SEQ ID NO: 12) to the fatty acid. An alternative embodiment has the following sequence: γE-CNTATCATQ-Orn-LAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is of the formula: 2 -γE-CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:12) to the fatty acid. Below is a depiction of compound XII using the standard single letter amino acid code, except that for glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, ornithine at position 11, lysine at position 26, and tyrosine at position 37, the structures of those amino acid residues have been expanded:

[0057] [ka] Compound 12; SEQ ID NO:12

[0058] Alternatively, the compound may have at least about 90% to about 99% sequence similarity to SEQ ID NO: 12. Alternatively, the compound may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 12.

[0059] One embodiment herein includes the following sequence: γE-CGTATCATG-Orn-LAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 (wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is represented by the formula: γE 2 -CO-(CH 2 ) 18 -CO 2H (SEQ ID NO: 13). An alternative embodiment herein is a γE-CGTATCATG-Orn-LAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 wherein there is a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, and the lysine at position 26 is of the formula: 2 -CO-(CH 2 ) 18 -CO 2 H (SEQ ID NO:13). Below is a depiction of compound XIII using the standard single letter amino acid code, except that the structures of the amino acid residues have been expanded for glutamic acid (γE) at position 1 (wherein the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, ornithine at position 11, lysine at position 26, and tyrosine at position 37:

[0060] [ka] Compound 13; SEQ ID NO:13

[0061] Alternatively, the compound may have at least about 90% to about 99% sequence similarity to SEQ ID NO: 13. Alternatively, the compound may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence similarity to SEQ ID NO: 13.

[0062] The present disclosure provides a compound consisting of SEQ ID NO:1, a compound consisting of SEQ ID NO:2, a compound consisting of SEQ ID NO:3, a compound consisting of SEQ ID NO:4, a compound consisting of SEQ ID NO:5, a compound consisting of SEQ ID NO:6, a compound consisting of SEQ ID NO:7, a compound consisting of SEQ ID NO:8, a compound consisting of SEQ ID NO:9, a compound consisting of SEQ ID NO:10, a compound consisting of SEQ ID NO:11, a compound consisting of SEQ ID NO:12, and / or a compound consisting of SEQ ID NO:13, or a pharma- ceutically acceptable salt thereof. The present disclosure also provides a pharmaceutical composition comprising a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, and one or more pharma- ceutically acceptable carriers, diluents, or excipients. The present disclosure also provides pharmaceutical compositions comprising a compound consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, and one or more pharma- ceutically acceptable carriers, diluents, or excipients.

[0063] The present disclosure further provides a compound comprising SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, or a pharma- ceutically acceptable salt thereof. In another embodiment, the present disclosure provides a compound consisting of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, or a pharma- ceutically acceptable salt thereof. Preferably, the compound of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25 comprises a disulfide or thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7. More preferably, the compound of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25 has a thioacetal bridge between the cysteine ​​at position 2 and the cysteine ​​at position 7, which is attached to a fatty acid linker moiety. Preferably, the fatty acid moiety is attached to a lysine.

[0064] The present disclosure provides a method of treating diabetes, obesity, dyslipidemia, and / or NASH in a patient, comprising administering to a patient in need of treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising same. The present disclosure further provides a method of reducing food intake, reducing body weight, reducing blood glucose, reducing HbA1c, and / or reducing triglycerides, comprising administering to a patient in need thereof an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising same. Preferably, the present disclosure provides a method of treating diabetes, obesity, dyslipidemia, and / or NASH in a patient, the method comprising administering to a patient in need of treatment an effective amount of a compound comprising SEQ ID NO: 2 or a pharmaceutical composition comprising same. More preferably, the present disclosure provides a method of treating diabetes in a patient, the method comprising administering to a patient in need of treatment an effective amount of a compound comprising SEQ ID NO: 2 or a pharmaceutical composition comprising same. Alternatively, the present disclosure provides a method of treating obesity in a patient, the method comprising administering to a patient in need of treatment an effective amount of a compound comprising SEQ ID NO: 2 or a pharmaceutical composition comprising same.

[0065] The present application provides a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in therapy. The present application further provides a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in the treatment of diabetes, obesity, dyslipidemia, and / or NASH. The present application also provides a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in therapy, treatment of diabetes, obesity, dyslipidemia, and / or NASH. More preferably, the present application provides a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in the treatment of diabetes. Alternatively, the present application provides a compound comprising SEQ ID NO: 2, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising same, for use in the treatment of obesity.Alternatively, the present application provides a compound comprising SEQ ID NO: 2, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising same, for use in the treatment of dyslipidemia.Alternatively, the present application provides a compound comprising SEQ ID NO: 2, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising same, for use in the treatment of NASH.

[0066] The present application provides a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, or a composition comprising same, for use in reducing food intake, reducing body weight, reducing blood glucose levels, reducing HbA1c, and / or reducing triglycerides. More preferably, the present application provides a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, or a composition comprising same, for use in reducing food intake, reducing body weight, reducing blood glucose levels, reducing HbA1c, and / or reducing triglycerides.

[0067] The present disclosure further provides for the use of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of diabetes, obesity, dyslipidemia, and / or NASH. Preferably, the present disclosure further provides for the use of a compound comprising SEQ ID NO:2, in the manufacture of a medicament for the treatment of diabetes, obesity, dyslipidemia, and / or NASH.

[0068] The present disclosure further provides the use of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for reducing food intake, reducing body weight, reducing blood glucose, reducing HbA1c, and / or reducing triglycerides. Preferably, the present invention further provides the use of a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for reducing food intake, reducing body weight, reducing blood glucose, reducing HbA1c, and / or reducing triglycerides.

[0069] 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 disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the amylin agonist peptides, pharmaceutical compositions, and methods, the preferred methods and materials are described herein.

[0070] Also, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that a plurality of elements is present, unless the context clearly requires that there is one and only one element. Thus, the indefinite article "a" or "an" normally means "at least one."

[0071] As used herein, "about" means within a statistically meaningful range of a value or values, such as, for example, a specified concentration, length, molecular weight, pH, sequence identity, time frame, temperature, volume, etc. Such values ​​or ranges may be within an order of magnitude, typically within 20%, more typically within 10%, and even more typically within 5% of a given value or range. The allowable variation encompassed by "about" will depend on the particular system under study and can be readily appreciated by one of ordinary skill in the art.

[0072] As used herein, with respect to one or more receptors, "activity," "activate," "activating," and the like, refer to the ability of a compound, such as a peptide herein, to bind to the receptor and induce a response at the receptor, as measured using assays known in the art, such as the in vitro assays described below.

[0073] As used herein, the term "amino acid" refers to both naturally occurring amino acids and non-coded amino acids. As is known in the art, an "amino acid" is a molecule characterized from a chemical standpoint by the presence of one or more amine groups and one or more carboxylic acid groups, and may contain other functional groups. Furthermore, amino acids are typically designated using standard one-letter codes (e.g., L is leucine), and α-methyl substituted residues of natural amino acids (e.g., α-methylleucine or αMeL, and α-methylphenylalanine or αMeF), as well as other specific non-coded amino acids, such as "γE", "NMeN", "NMeD", "Orn", etc. The structures of the non-coded amino acids of the present disclosure are shown below:

[0074] [ka]

[0075] As used herein, "γE" means γ-glutamic acid (except that the peptide bond is formed using the side chain carboxylic acid group in the γ position rather than the typical α position). As used herein, "Orn" means L-ornithine. As used herein, "αMeL" means α-methyl-L-leucine. As used herein, "αMeF" means α-methyl-L-phenylalanine. As used herein, "NMeN" means N-methyl-asparagine and "NMeD" means N-methyl-aspartic acid.

[0076] As used herein, "AEEA" means 2-[2-(2-amino-ethoxy)-ethoxy]acetic acid, the structure of which is as follows:

[0077] [ka]

[0078] As used herein, "amylin agonist peptide" means a compound, such as a synthetic peptide or polypeptide, that activates a target receptor and elicits at least one in vivo or in vitro effect elicited by a natural agonist for that receptor.

[0079] As used herein, "chronic weight management" refers to a method of maintaining weight loss as an adjunct to a reduced calorie diet and increased physical activity in humans currently or previously characterized as obese.

[0080] The term "diabetes" refers to a disease in which the body's ability to produce or respond to the hormone insulin is impaired, resulting in abnormal metabolism of carbohydrates and elevated levels of glucose in the blood and urine. As used herein, the term "diabetes" can refer to a chronic condition that affects the way the body processes blood sugar or glucose, e.g., type 2 diabetes mellitus (T2DM), a chronic condition in which the pancreas produces little or no insulin, e.g., type 1 diabetes mellitus (T1DM), a condition in which blood glucose levels are high but not high enough to be type 2 diabetes, e.g., prediabetes, a form of hyperglycemia that affects pregnant women, e.g., gestational diabetes.

[0081] The term "dyslipidemia" refers to a disorder of lipoprotein metabolism, including overproduction or deficiency of lipoproteins. Dyslipidemia may be manifested in elevated total cholesterol, low-density lipoprotein (LDL) cholesterol and triglyceride concentrations, and / or decreased high-density lipoprotein (HDL) concentrations in the blood. Dyslipidemia may or may not occur in association with diabetes.

[0082] As used herein, a "fatty acid" consists of a linear chain of an even number of carbon atoms with hydrogen atoms along the length of the chain and a carboxyl group (-COOH) at one (mono-acid) or both (diacid) ends of the chain. In a preferred embodiment, the "fatty acid" moiety is 20 It is a diacid.

[0083] [ka]

[0084] As used herein, "thioacetal bridge" refers to the organic synthesis of a thioacetal group to functionally recrosslink a disulfide bond(s) within a peptide structure. 2 -S-) is shown below with a methylene group inserted between the two sulfur atoms.

[0085] [ka]

[0086] As used herein, the term "treatment" or "treating" refers to the management and care of a patient having a condition for which administration of an amylin receptor peptide agonist is indicated to combat or alleviate the symptoms and complications of the condition. Treating includes administering one of the compounds disclosed herein or a pharmaceutical composition containing the same to a patient in need of the compound or a pharmaceutical composition containing the same to prevent the onset of the symptoms or complications, to alleviate the symptoms or complications, or to eliminate a disease, condition, or disorder. Preferably, treating includes administering a compound of the present disclosure or a pharmaceutical composition containing the compound of the present disclosure to a patient in need of a compound of the present disclosure or a pharmaceutical composition containing the compound of the present disclosure to result in a net reduction in body weight, reduced food intake, reduced blood glucose levels, reduced HbA1c, and / or reduced triglyceride levels. The patient to be treated is a mammal, preferably a human.

[0087] As used herein, the term "effective amount" refers to an amount or dose of one or more of the compounds described herein that, upon single or multiple administration to an individual in need thereof, provides the desired effect in such an individual during diagnosis or treatment (i.e., can produce a clinically measurable difference in the individual's condition, such as, for example, a net reduction in body weight, a reduction in food intake, a reduction in blood glucose levels, a reduction in HbA1c, and / or a reduction in triglyceride levels). Effective amounts can be readily demonstrated by those skilled in the art by the use of known techniques and by observing results obtained under similar circumstances. In determining an effective amount for an individual, numerous factors are taken into account, including, but not limited to, the species of mammal, its size, age, and general health, the particular disease or disorder involved, the involvement or severity of the disease or disorder, the individual's response, the particular compound administered, the mode of administration, the bioavailability characteristics of the administered preparation, the selected dosage, the use of concomitant drugs, and other relevant circumstances. Preferably, an effective amount of the disclosed compound or pharmaceutical composition containing the disclosed compound administered to a patient in need thereof will result in a net reduction in body weight, reduced food intake, reduced blood glucose levels, reduced HbA1c, and / or reduced triglyceride levels. Dosages can include a higher initial loading dose followed by smaller doses. Effective doses of the compounds provided herein can be 0.05 μg / kg to 5000 μg / kg, or 0.01 nmol / kg to 1000 nmol / kg.

[0088] As used herein, "half-life" or "t 1 / 2 " refers to the time it takes for half of the amount of a compound, such as a peptide, herein to be removed by biological processes from a fluid, such as serum or plasma, or other physiological space of an individual. Alternatively, t 1 / 2 It can also mean the time it takes for such an amount of peptide to lose half of its pharmacological, physiological, or radiological activity.

[0089] As used herein, "median effective concentration" or "EC 50 " means the concentration of a compound that results in 50% activation / stimulation of an assay endpoint, such as a dose-response curve.

[0090] As used herein, "long-acting" means that the binding affinity and activity of the compositions herein continues for a longer period of time than the native peptide or protein, thereby allowing dosing at least as frequently as once a day, or even three times a week, twice a week, once a week, or even once a month. The time-action profile of the compounds herein may be measured using known pharmacokinetic testing methods, such as those described in the Examples below.

[0091] The term "NASH" refers to non-alcoholic steatohepatitis, also known as fatty liver disease. "NASH" also refers to inflammation and damage of the liver caused by fat accumulation in the liver. "NASH" also refers to a subtype of non-alcoholic fatty liver disease ("NAFLD"). In some embodiments, "NASH" can be synonymous with "NAFLD".

[0092] As used herein, the term "obesity" refers to a disorder involving excess body fat that increases the risk of health problems. The term "obesity" also refers to a body weight that is greater than what is considered healthy for a given height. The term "obesity" also refers to a BMI greater than 30.0, or a BMI of 27.0 or greater (overweight) with at least one weight-related comorbid condition (e.g., hypertension, type 2 diabetes mellitus, or dyslipidemia).

[0093] As used herein, body mass index (BMI) refers to a person's weight in kilograms divided by the square of their height in meters.

[0094] As used herein, "patient" and "individual" are used interchangeably and refer to a mammal, preferably a human. In certain embodiments, the patient, preferably a human, is further characterized by a disease, disorder, or condition that may benefit from the administration of a compound that stimulates both amylin and calcitonin receptors.

[0095] In some embodiments, pharmaceutical compositions comprising the compounds disclosed herein may be administered orally to a patient in need of such treatment. Pharmaceutical compositions comprising the compounds disclosed herein may be administered parenterally to a patient in need of such treatment. Parenteral administration may be performed by subcutaneous, intramuscular, or intravenous injection using a syringe, optionally a pen-like syringe, or a mechanically driven injector. Alternatively, parenteral administration may be performed using an infusion pump. An embodiment of the compounds disclosed herein provides a pharmaceutical composition suitable for administration to a patient comprising administering a therapeutically effective amount of a compound disclosed herein and one or more pharma- ceutically acceptable excipients to a patient in need of the pharmaceutical composition. Such pharmaceutical compositions may be prepared by any of a variety of techniques using conventional excipients for pharmaceutical products well known in the art. (Remington's Pharmaceutical Sciences, 21st Edition, University of the Sciences in Philadelphia, Philadelphia, PA, USA (2006)).

[0096] As used herein, "sequence similarity" refers to a quantitative characteristic of two or more nucleic acid or amino acid sequences of biological compounds, such as, for example, the match over the entire length or comparison window of two or more sequences. Sequence similarity can be measured by (1) percent identity or (2) percent similarity. Percent identity measures the percentage of identical residues between two biological compounds divided by the length of the shortest sequence, while percent similarity measures identity and also includes sequence gaps and residue similarity in the assessment. Methods and algorithms for determining sequence similarity are well known in the art and need not be described exhaustively here. A particular percentage of identical nucleotides or amino acid positions is at least about 75%, 80%, 85%, 86, 76, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, or more.

[0097] The compounds disclosed herein may be used in combination, simultaneously, separately or sequentially, with one or more additional therapeutic agents useful for inducing weight loss, treating diabetes, diabetes-related conditions, obesity and / or chronic weight management, dyslipidemia and / or NASH. Non-limiting examples of additional therapeutic agents that may be combined with the claimed compounds include insulin or insulin analogs; biguanides; sulfonylureas; thiazolidinediones; dipeptidyl peptidase-4 ("DPP-4") inhibitors; sodium-dependent glucose transporter (SGLT2) inhibitors; incretin compounds such as glucagon-like peptide-1 (GLP-1) or GLP-1 analogs, gastric inhibitory polypeptide (GIP) or GIP analogs, oxyntomodulin or oxyntomodulin analogs; growth differentiation factor 15 (GDF15) agonist compounds; peptide YY (PYY) analogs; GIP / GLP-1 dual agonists; Gcg / GIP / GLP-1 triagonists (triagonists of glucagon, GIP, and GLP-1); or a combination of any of the above agents. The claimed compounds and the additional therapeutic agent(s) can be co-administered via the same delivery route and device, such as a single pill, capsule, tablet, or injectable formulation, or can be administered separately at the same time in separate delivery devices or routes, or can be administered sequentially.

[0098] Another embodiment of the present disclosure is a method of treating a condition selected from the group consisting of diabetes, obesity, NASH, and / or dyslipidemia in a patient in need of such treatment, comprising administering to the patient an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with an effective amount of an incretin or incretin analog. Preferably, an embodiment of the present disclosure is a method of treating a condition selected from the group consisting of diabetes, obesity, NASH, and / or dyslipidemia in a patient in need of such treatment, comprising administering to the patient an effective amount of a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, in combination with an effective amount of an incretin or incretin analog.

[0099] A particular embodiment of the present disclosure is a method of treating a condition selected from the group consisting of diabetes, obesity, NASH, and / or dyslipidemia in a patient in need of such treatment, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with a GLP-1 agonist. A particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, or a pharma- ceutically acceptable salt thereof, in combination with a CLP-1 agonist, comprising compound XVII (SEQ ID NO:18). Another particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, in combination with a CLP-1 agonist comprising compound XVII (SEQ ID NO:18).

[0100] A particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with an analog of oxyntomodulin. A particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with an analog of oxyntomodulin, including compound XVIII (SEQ ID NO:19). Another specific embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO: 2, or a pharma- ceutically acceptable salt thereof, in combination with an analog of oxyntomodulin, comprising compound XVIII (SEQ ID NO: 19).

[0101] One particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with a triagonist of glucagon, GIP, and GLP-1. A particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with a triagonist of glucagon, GIP, and GLP-1, including compound XIX (SEQ ID NO:20). Another particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, in combination with a triagonist of glucagon, GIP, and GLP-1, including compound XIX (SEQ ID NO:20).

[0102] A particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with a GIP / GLP-1 dual agonist. A particular embodiment of the present disclosure is a method of treating a patient with diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, in combination with a GIP / GLP-1 dual agonist comprising SEQ ID NO:21. Another particular embodiment of the present disclosure is a method of treating a patient having diabetes, obesity, NASH, and / or dyslipidemia, comprising administering to a patient in need of such treatment an effective amount of a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, in combination with a dual GIP / GLP-1 agonist, comprising SEQ ID NO:21.

[0103] Another embodiment of the disclosure is a compound comprising SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, or SEQ ID NO:14, or a pharma- ceutically acceptable salt thereof, for use in separate, simultaneous, or sequential combination with an incretin or incretin analog for the treatment of diabetes, obesity, NASH, and / or dyslipidemia. Preferably, an embodiment of the disclosure is a compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof, for use in separate, simultaneous, or sequential combination with an incretin or incretin analog for the treatment of diabetes, obesity, NASH, and / or dyslipidemia. More preferably, the incretin or incretin analog is a GIP / GLP-1 dual agonist comprising SEQ ID NO:21.

[0104] Certain abbreviations used herein are defined as follows: "ACN" refers to acetonitrile, "AMY1R" refers to amylin receptor 1, "cAMP" refers to cyclic adenosine monophosphate, "CT" refers to calcitonin, "DCM" refers to dichloromethane, "DIEA" refers to diisopropylethylamine, "DMF" refers to N,N-dimethylformamide, "DMSO" refers to dimethylsulfoxide, "DODT" refers to 2,2'-(ethylenedioxy)diethanethiol, "FBS" refers to fetal bovine serum, "Fmoc" refers to fluorenylmethyloxycarbonyl, "GDF15" refers to growth differentiation factor 15, "GPCR" refers to G protein-coupled receptor, "HEPES" refers to 4-(2-hydroxyethyl)phenylalanine, and "HEPES" refers to 4-(2-hydroxyethyl)phenylalanine. "Methoxy" refers to tris(2-carboxyethyl)-1-piperazineethanesulfonic acid, "HTRF" refers to homogeneous time-resolved fluorescence, "IBMX" refers to 1-methyl-3-isobutylxanthine, "MEM" refers to minimum essential medium, "Mtt" refers to 4-methyltrityl, "NASH" refers to non-alcoholic fatty liver disease, "NEAA" refers to non-essential amino acids, "PyAOP" refers to (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate, "RP-HPLC" refers to reversed-phase high pressure liquid chromatography, "TCEP" refers to tris(2-carboxyethyl)phosphine hydrochloride, "TFA" refers to trifluoroacetic acid, and "TRIS" refers to tris(hydroxymethyl)aminomethane. EXAMPLES

[0105] Example 1: Preparation and purification of Compound I and Compound II Compound I and Compound II are prepared according to the following steps: First, compound I (SEQ ID NO: 1) is synthesized using fluorenylmethyloxycarbonyl (Fmoc) / tert-butyl (t-Bu) chemistry on a Symphony 12-channel multiple peptide synthesizer (Protein Technologies, Inc., Tucson, Arizona).

[0106] Polystyrene Rink Amide MBHA Resin LL resin (Novabiochem, sub: 0.35 meq / g, 100-200 mesh, catalog number 855045) is used for the synthesis on a 0.13 mmol scale. Standard side chain protecting groups are used. Boc-Glu-OtBu is used at position 1. Fmoc-Lys(Mtt)-OH is used at lysine position 26. The Fmoc group is removed using 20% ​​piperidine in DMF before each coupling step (2 times 7 min). All amino acid couplings are carried out at 60° C. for 30 min at a 7.7-fold molar excess over theoretical peptide loading using equimolar ratios of Fmoc amino acid (0.3 M), diisopropylcarbodiimide (0.9 M), and Oxyma (0.9 M). Subsequent amino acid couplings of αMeF at position 15 and NMeN at position 22 are carried out at 60° C. for 3 and 6 h, respectively. For compound I (SEQ ID NO:1), the resin is treated with a cleavage cocktail at this point (conditions are described after the procedure for adding the fatty acid-linker moiety to generate compound II). Below is a schematic depiction of compound I (SEQ ID NO:1) using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (the peptide bond is formed with the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeN at position 22, and tyrosine at position 37:

[0107] [ka] Compound I; SEQ ID NO:1

[0108] The resin is then thoroughly washed six times with DCM to remove residual DMF. The Mtt protecting group of lysine at position 26 is selectively removed from the peptide-resin using two treatments of 30% hexafluoroisopropanol (Oakwood Chemical) in DCM (2 x 40 min treatments). Subsequent attachment of the fatty acid linker moiety is achieved by coupling Fmoc-glutamic acid α-t-butyl ester (Fmoc-Glu-OtBu (Ark Pharm, Inc.)) and mono-OtBu-eicosanoic acid (WuXi AppTec, Shanghai, China). A three-fold excess of reagents (AA:PyAOP:DIEA=1:1:1 mol / mol) is used for each coupling with a duration of 1 hour.

[0109] Once synthesis is complete, wash the peptide resin with DCM and air dry thoroughly. The dried resin is incubated at room temperature for 2 h with 10 mL of cleavage cocktail (TFA:DODT:TIS:H 2 The resin is filtered off, washed twice with 2 mL of neat TFA each, and the combined filtrates are treated with 4 portions of cold diethyl ether (-20°C) to precipitate the crude peptide. The peptide / ether suspension is then centrifuged at 3500 rpm for 2 min to form a solid pellet, the supernatant is decanted, and the solid pellet is triturated with ether two more times. To create the thioacetal crosslink, the air-dried pellet is dissolved in 5 mL of 20 mM potassium phosphate buffer and 3 mL of ACN. Once completely dissolved, HCl is added to the eluate with good stirring. 2 Add TCEP (30 μM, 5 equiv.), diiodomethane (8 equiv.), and triethylamine (10 equiv.) in HO and stir for 5-30 min. Monitor the reaction using LCMS. It is usually complete in 5 min. After the reaction is complete, wash the mixture with 1 mL of HO with 0.1% TFA. 2 Add 200 to the mixture along with 6 mL of acetic acid. If the solution is cloudy, add more ACN as needed.

[0110] The crude peptide is purified by RP-HPLC on a Phenomenex PhenylHexyl column (5 μm, 100A, 250×21.2 mm, part number: 00G-4257-P0-AX) using a linear gradient with 100% acetonitrile and 0.1% TFA / water buffer system. The purity of the peptide is assessed using analytical RP-HPLC using a Waters Symmetry Shield RP18 column (3.5 μm, 6×100 mm, part number: 186000179) with a pool standard of >95%. The main pool purity of compound II (SEQ ID NO:2) is found to be >98.4%. Subsequent lyophilization of the final main product pool results in the lyophilized peptide TFA salt. The molecular weight of compound I (SEQ ID NO:1) is determined by LC / MS (observed: [M+3H] 3+ = 1315.2; calculated value [M+3H] 3+ = 1315.5; MW(average) found = 3942.6; MW(average) calculated = 3943.4). The molecular weight of compound II (SEQ ID NO:2) is determined by LC / MS (Found: [M+3H] 3+ =1509.5;Calculated value [M+3H] 3+ = 1509.7; MW(average)observed = 4525.5; MW(average)calculated = 4526.1. Below is a schematic depiction of compound II (SEQ ID NO:2) using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (the peptide bond is formed with the side chain carboxylic acid at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeN at position 22, lysine at position 26, and tyrosine at position 37:

[0111] [ka] Compound II; SEQ ID NO:2

[0112] Methods similar to those described above and known to those of skill in the art can be used to synthesize the peptide backbone, conjugate the fatty acid-linker moiety, check the purity, and confirm the molecular weight of the compounds of the invention described herein.

[0113] Example 2: Preparation and purification of Compound III and Compound IV Compound III and Compound IV are prepared according to the process outlined in Example 1.

[0114] Below is a schematic depiction of compound III (SEQ ID NO:3) using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (where the peptide bond is formed with the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, and tyrosine at position 37:

[0115] [ka] Compound III; SEQ ID NO:3

[0116] The molecular weight of compound III (SEQ ID NO:3) is determined by LC / MS (observed value: [M+3H] 3+ =1314.9;Calculated value [M+3H] 3+ = 1315.8; MW(average) actual = 3941.7; MW(average) calculated = 3944.4).

[0117] Below is a schematic depiction of compound IV (SEQ ID NO:4) using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (where the peptide bond is formed with the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, lysine at position 26, and tyrosine at position 37:

[0118] [ka] Compound IV; SEQ ID NO:4

[0119] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1509.7;Calculated value [M+3H] 3+ = 1510.0; MW(average) actual = 4526.1; MW(average) calculated = 4527.1).

[0120] Example 3: Preparation and purification of Compound V and Compound VI Compound V and Compound VI are prepared according to the process outlined in Example 1.

[0121] Below is a schematic depiction of Compound V (SEQ ID NO:5) using the standard single letter amino acid code, except that the structures of the amino acid residues Cysteine ​​at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, and Tyrosine at position 37 have been expanded:

[0122] [ka] Compound V; SEQ ID NO:5

[0123] The molecular weight of compound V (SEQ ID NO: 5) is determined by LC / MS (observed value: [M+3H] 3+ =1314.9;Calculated value [M+3H] 3+ = 1315.5; MW(average) actual = 3941.7; MW(average) calculated = 3943.4).

[0124] Below is a schematic depiction of compound VI (SEQ ID NO:6) using standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: Lysine at position 1, Cysteine ​​at positions 2 and 7, Orn at position 11, αMeF at position 15, NMeD at position 22, and Tyrosine at position 37:

[0125] [ka] Compound VI: SEQ ID NO:6

[0126] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1509.4;Calculated value [M+3H] 3+ = 1509.7; MW(average) actual = 4525.2; MW(average) calculated = 4526.1).

[0127] Example 4: Preparation and purification of Compound VII and Compound VIII Compounds VII and VIII are prepared according to the methods outlined in Example 1.

[0128] Below is a schematic depiction of compound VII (SEQ ID NO:7) using the standard single letter amino acid code, except that the structures of the amino acid residues Cysteine ​​at positions 2 and 7, Orn at position 11, αMeL at position 12, and Tyrosine at position 37 have been expanded:

[0129] [ka] Compound VII; SEQ ID NO:7

[0130] The molecular weight of compound VII (SEQ ID NO: 7) is determined by LC / MS (observed value: [M+3H] 3+ =1317.9;Calculated value [M+3H] 3+ = 1318.1; MW(average) actual = 3950.7; MW(average) calculated = 3951.4).

[0131] Below is a schematic depiction of compound VIII (SEQ ID NO:8) using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: Lysine at position 1, Cysteine ​​at positions 2 and 7, Orn at position 11, αMeL at position 12, and Tyrosine at position 37:

[0132] [ka] Compound VIII; SEQ ID NO:8

[0133] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1512.2;Calculated value [M+3H] 3+ = 1512.4; MW(average) actual = 4533.6; MW(average) calculated = 4534.2).

[0134] Example 5: Preparation and purification of compound IX Compound IX is prepared according to the process outlined in Example 1.

[0135] Below is a schematic depiction of compound IX (SEQ ID NO:9) using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, lysine at position 26, and tyrosine at position 37:

[0136] [ka] Compound IX; SEQ ID NO:9

[0137] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1558.3;Calculated value [M+3H] 3+ = 1558.8; MW(average) actual = 4671.9; MW(average) calculated = 4673.3).

[0138] Example 6: Preparation and purification of compound X Compound X is prepared according to the process outlined in Example 1.

[0139] Below is a schematic depiction of compound X (SEQ ID NO:10) using the standard single letter amino acid code, except that the structures of the amino acid residues glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, lysine at position 26, and tyrosine at position 37 have been expanded:

[0140] [ka] Compound X; SEQ ID NO: 10

[0141] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1558.5;Calculated value [M+3H] 3+ = 1558.8; MW(average) actual = 4672.5; MW(average) calculated = 4673.3).

[0142] Example 7: Preparation and purification of compound XI Compound XI is prepared according to the process outlined in Example 1.

[0143] Below is a schematic depiction of compound XI (SEQ ID NO:11) using the standard single letter amino acid code, except that for glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, lysine at position 26, and tyrosine at position 37, the structures of those amino acid residues have been expanded:

[0144] [ka] Compound XI; SEQ ID NO:11

[0145] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1553.2;Calculated value [M+3H] 3+ = 1553.4; MW(average) actual = 4656.6; MW(average) calculated = 4657.2).

[0146] Example 8: Preparation and purification of compound XII Compound XII is prepared according to the process outlined in Example 1.

[0147] Below is a schematic depiction of compound XII (SEQ ID NO:12) using the standard single letter amino acid code, except that the structures of the following amino acid residues have been expanded: glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, lysine at position 26, and tyrosine at position 37:

[0148] [ka] Compound XII; SEQ ID NO:12

[0149] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1577.4;Calculated value [M+3H] 3+ = 1577.8; MW(average) actual = 4729.2; MW(average) calculated = 4730.3).

[0150] Example 9: Preparation and purification of compound XIII Compound XIII is prepared according to the process outlined in Example 1.

[0151] Below is a schematic depiction of compound XIII (SEQ ID NO:13) using the standard single letter amino acid code, except that for glutamic acid (γE) at position 1 (where the peptide bond is formed using the side chain carboxylic acid group at the γ position rather than the typical α position), cysteines at positions 2 and 7, Orn at position 11, lysine at position 26, and tyrosine at position 37, the structures of those amino acid residues have been expanded:

[0152] [ka] Compound XIII; SEQ ID NO:13

[0153] The molecular weight is determined by LC / MS (measured value: [M+3H] 3+ =1481.2;Calculated value [M+3H] 3+ = 1481.3; MW(average) actual = 1440.6; MW(average) calculated = 4441.0).

[0154] Preparation of disulfide bond cross-links To create disulfide bonds, prepare the oxidation solution by adding 10 drops of iodine solution (2% iodine in AcOH) to 40 mL of 20% MeCN / 20% AcOH / 60% water in a flask. Separately, make a peptide solution by dissolving the air-dried peptide pellet in 5 mL of AcOH. Add the peptide solution dropwise to the oxidation solution while stirring. Add additional iodine solution (2% iodine in AcOH) as needed to maintain a light yellow solution in the flask. After all the peptide solution has been added, the light brown / yellow color is maintained for 5 minutes. Excess iodine is neutralized by adding 1 drop of saturated ascorbic acid to the oxidation solution. Filter the solution through a 0.45 μm filter and the peptide is ready for purification.

[0155] Example 10: In vitro functional activity of amylin agonist peptides The AMY1 and CT receptors are GPCRs that are functionally coupled to the Gαs protein. Stimulation of these receptors increases the production of intracellular cAMP, which can be detected using standard in vitro techniques. The in vitro activity of the peptides is measured by the amount of cAMP formed in human AMY1R and CTR overexpressing cells.

[0156] Human CT receptor is stably expressed in human bladder cells (UM-UC-3 or UMUC3) under the control of pcDNA expression vector. UMUC3 cell line is cultured in MEM 1X (Meditech Inc., 17-305-CV) supplemented with 10% FBS, 1% antibiotic / antimycotic solution, 1 mM sodium pyruvate, 1XMEM NEAA, 1XGlutaMAX-I. Human CTa-pcDNA3.1 Hygro(+)(T2616)DNA plasmid is transfected into UMUC3 cells using LipofectAMINE 2000 transfection reagent (Invitrogen, 11668-019). After 20 days under selection, mRNA levels from different clones are measured to confirm the expression of hCTR. To determine the function of overexpressed hCTR cells, intracellular cAMP levels in response to salmon calcitonin are measured and compared to the expression of hCTR mRNA in each clone.

[0157] Human AMY1R stable cell line is generated by further transfecting human RAMP1-pCMVpuroPB(T14213) into hCTR clone cells of UMUC3. After selection, the mRNA level of human RAMP1 from different clones is measured to confirm the expression of human AMY1R.

[0158] hAMY1R cells are cultured in MEM 1X (Corning) supplemented with 10% FBS, 1% antibiotic / antimycotic solution, 1 mM sodium pyruvate, 1X MEM NEAA, 1X GlutaMAX-I, 200 μg / mL hygromycin B, and 0.4 μg / mL puromycin. hCTR cells are cultured in the same medium lacking puromycin. Cultured cells are grown to 70% confluency and then incubated overnight in fresh medium.

[0159] On the day of the assay, 10 μL of assay buffer (MEM without phenol red (Corning, Cat. No. 17-305-CV), 0.1% casein, 0.5 mM IBMX, 5 mM HEPES, pH 7.4) is dispensed into each well of a white poly-D-lysine coated 384-well plate (Corning, Cat. No. 354661). Peptides diluted in DMSO are added (200 nL / well) in a 1:3 dilution series using an ECHO acoustic liquid handler (Beckman). Cultured cells are detached with TrypLE Express (Gibco), resuspended in assay buffer, and 10 μL containing 1200 cells / well (hCTR) or 1500 cells / well (hAMY1R) is dispensed into each well. Plates are incubated at room temperature for 1 hour.

[0160] The amount of intracellular cAMP is quantified using HTRF technology (Homogeneous Time-Resolved Fluorescence, Cisbio) according to the manufacturer's instructions. Briefly, 10 μL of cAMP-d2 conjugate and 10 μL of anti-cAMP-cryptate conjugate in lysis buffer are incubated with treated cells for 60 min at room temperature. The HTRF signal is immediately detected using an Envision plate reader (Perkin-Elmer) and the ratio of fluorescence at 665 nm vs. 620 nm is calculated. The raw data is converted to cAMP amount (pmol / well) using a cAMP standard curve generated for each experiment. Relative EC 50 Values ​​are calculated from the upper and lower ranges of the concentration-response curves defined with a four-parameter logistic curve fitting program (Genedata Screener® v12.0.4) using 1 nM Salmon CT (Bachem) as the maximum and buffer alone as the minimum. Compounds of the present application exhibit selective activity at amylin versus calcitonin receptors, as shown in Table 1.

[0161] Table 1. Comparison of functional activity data for amylin and calcitonin receptors [Table 1]

[0162] Example 11: In vitro binding affinity of amylin agonist peptides to the hAMY1 and hCT receptors Membranes from human AMY1R and CTR overexpressing cells (described in Example 10) were isolated by standard methods and used for binding assays. The equilibrium dissociation constants (Kd) of the various receptor / radioligand interactions are determined from saturation binding analysis using the same reagents and buffers as described below for compound testing. The Kd values ​​measured for the receptor preparations used in this study are as follows: hAMY1R: 0.067 nM; human calcitonin receptor (hCTR): 0.046 nM.

[0163] hAMY1R binding protocol Receptor binding affinities (Ki) of rAMY, hCT, and hAMY agonist peptides are determined from competitive radioligand binding filter assays. The assay buffer is 50 mM HEPES, pH 7.1, 5 mM MgCl 2, 5 mM KCl, 0.2% (w / v) bacitracin, 0.003% (w / v) saponin, used to dilute radioligand and membrane preparations. Binding reactions are performed in a total reaction volume of 0.1 mL in a polystyrene 96 deep-well assay block. Iodinated rat amylin (custom synthesis by ViTrax; 2200 Ci / mmol; 125I-rAMY) is initially diluted to a concentration of approximately 50 pM in assay buffer. Test compounds and nonspecific binding (NSB: defined as 300 nM rAMY) are added to aliquots of radioactivity buffer. Briefly, test compounds are diluted to an initial concentration of 200 nM and serially diluted in 4-fold steps in 125I-rAMY, then 0.05 mL of diluted test compound, NSB, or total binding (defined as neat 125I-rAMY) is transferred to a 96-well polystyrene assay block. The binding reaction is initiated by adding 0.05 mL of 200 μg / mL hAMY1R diluted in assay buffer to the radioactivity buffer. The assay block is gently vortexed, sealed with parafilm, and incubated at room temperature for approximately 20 hours. Thirty minutes before the incubation is complete, filter mats (Perkin Elmer printed filter mats, catalog number 1450-421) are immersed in a solution consisting of saponin-free assay buffer, but supplemented with 0.1% (w / v) fatty acid-free bovine serum albumin (FAF-BSA) and 0.5% (v / v) polyethyleneimine (PEI). At the end of the incubation, bound ligand is separated from free ligand by the addition of ice-cold quench buffer consisting of 100 mM NaCl, 50 mM TRIS-HCl, pH 7.1, and immediately harvested onto the filter mats by vacuum filtration using a TomTec 96-well filter harvester. The following filter protocol was used: dry time, 7; cycle 3 repetitions, 0; wash time, 1; soak time, 1; 1st aspiration time, 4; wash / aspiration time, 7; 2nd aspiration time, 4; air pressure, 2 PSI. The final assay concentration range of the peptides tested in the response curves is 100 nM to 0.00038 nM.

[0164] hCTR binding protocol The receptor binding affinity (Ki) of rAMY, hCT, and hAMY agonist peptides on hCTR membranes is determined from a competitive radioligand binding filter assay. The assay procedure is similar to the hAMY1R binding assay, except that 125I-hCT (custom synthesis by ViTrax; 2200 Ci / mmol) is used as the hot ligand. Binding reactions are performed in a total reaction volume of 0.2 mL with 14 pM 125I-hCT and 20 μg / mL hCTR membranes and incubated for 20 hours. The final assay concentration range of the tested peptides in the response curve is 2500 nM to 0.00128 nM. The compounds of the present application show selective activity at amylin receptors versus calcitonin receptors, as shown in Table 2.

[0165] Table 2. In vitro binding (Ki) at human amylin and calcitonin receptors [Table 2]

[0166] Example 12: In vivo effects on food intake and body weight in normal rats Male Sprague Dawley rats, obtained from Envigo RMS, Indianapolis, IN, were maintained on chow (2014; Teklad Global (Envigo RMS, Indianapolis, IN), single-housed in a temperature-controlled facility (72.0°F; 22.2°C) with a 12:12-h reversed light cycle (off at 10 am) and allowed free access to food and water. At 10 weeks of age, non-fasted body weights and initial food weights were recorded and animals were administered a single subcutaneous injection (SC) of vehicle or acylated peptide (1 mL / kg), after which body weights and food intakes were measured daily for 4 days post-dosing. Area under the curve (AUC) analysis was calculated for both body weight and food intake relative to vehicle.

[0167] Table 3: Changes in body weight and food intake over 4 days in Sprague Dawley rats following a single dose of long-acting amylin agonist peptides. [Table 3] * Mean body weight change at 96 hours post-dose compared to initial body weight of untreated animals. ** Cumulative food intake of peptide-treated animals at 96 hours post-dose versus vehicle-treated animals at 96 hours post-dose.

[0168] Example 13: In vivo effects on food intake and body weight in diet-induced obese rats Male Long Evans rats, 14 weeks of age, obtained from Envigo RMS, Indianapolis, IN, were fed a high-fat diet (40% of caloric intake was fat, TD.95217 (Envigo RMS, Indianapolis, IN)) until 35-45 weeks of age. Animals were individually housed in a temperature-controlled (75.0°F; 23.9°C) facility with a 12-h reversed light / dark cycle (lights off at 10:00 AM) and had food and water available ad libitum. Body weight was recorded and body composition (fat mass) was determined using quantitative nuclear magnetic resonance analysis (ECHO MRI, 3-1 Composition Analyzer; Echo Medical Systems, Houston, TX), followed by randomization into experimental groups (n=5). Rats were injected subcutaneously with vehicle or peptide (1 mL / kg) every 3 days (days 1, 4, 7, 10, and 13). Daily body weights are recorded and changes at the end of treatment (day 14) are calculated as a percentage of pre-treatment body weight (day 1). Body composition is measured on day 14 and changes in fat and lean mass (body weight minus fat mass) are calculated as change in grams from pre-treatment values.

[0169] Table 4: Change in body weight in a 2 week study in diet-induced obese rats using amylin agonist peptides. [Table 4] * Change in mean body weight or fat or lean mass at week 2 compared to initial body weight of untreated animals.

[0170] Example 14: Pharmacokinetics in male Sprague Dawley rats The pharmacokinetics of Compound II (SEQ ID NO:2), Compound IV (SEQ ID NO:4), Compound VI (SEQ ID NO:6), and Compound VIII (SEQ ID NO:8) are evaluated following a single subcutaneous dose of 30 nmol / kg in male Sprague Dawley rats. Blood samples are collected at 1, 3, 6, 12, 24, 48, 72, and 96 hours after SC administration. The individual plasma concentrations obtained are used to calculate pharmacokinetic parameters. Peptide plasma (K 3 EDTA) concentrations are determined using a qualified LC / MS method that measures the intact mass of the peptides. Each peptide and analog is extracted from plasma as an internal standard. A high-resolution Thermo Q-Exactive is used for LC / MS detection. Mean pharmacokinetic parameters are shown in Table 5.

[0171] Table 5. Mean pharmacokinetic parameters of peptides following a single subcutaneous dose of 30 nmol / kg to male Sprague Dawley rats. [Table 5] Abbreviation:T 1 / 2 = half-life, T max = time to reach maximum concentration, C max / D = maximum plasma concentration divided by dose, AUCINF / D = AUCinf divided by dose, CL / F = clearance / bioavailability. Note: Data are means, n=3 per group.

[0172] Example 15: Pharmacokinetics in male cynomolgus monkeys The pharmacokinetics of Compound II (SEQ ID NO:2), Compound IV (SEQ ID NO:4), Compound VI (SEQ ID NO:6), and Compound VIII (SEQ ID NO:8) are evaluated following a single subcutaneous dose of 20 nmol / kg to male cynomolgus monkeys. Blood samples are collected at the following time points: 1, 3, 6, 12, 24, 48, 72, 120, 168, 240, 336, 408, and 504 hours after SC administration. The resulting individual plasma concentrations are used to calculate pharmacokinetic parameters. Peptide plasma (K 3 EDTA) concentrations are determined using a qualified LC / MS method that measures the intact mass of the peptides. Each peptide and analog is extracted from plasma as an internal standard. A high-resolution Thermo Q-Exactive is used for LC / MS detection. Mean pharmacokinetic parameters are shown in Table 6.

[0173] Table 6. Mean pharmacokinetic parameters of peptides following a single subcutaneous dose of 20 nmol / kg to cynomolgus monkeys. [Table 6] Abbreviation:T 1 / 2 = half-life, T max = time to reach maximum concentration, C max / D = maximum plasma concentration divided by dose, AUCINF / D = AUCinf divided by dose, CL / F = clearance / bioavailability. Note: Data are means, n=3 per group.

[0174] Example 16: Immunogenicity Risk Assessment Dendritic cell (DC) internalization assay This assay evaluates the ability of human DCs to internalize a test antibody. CD14+ cells are cultured and differentiated into immature DCs with IL-4 and GM-CSF. Test antibodies, isotype controls, or positive controls are pre-incubated with detection agent (Fab-QSY7-TAMRA) at a 1:1 ratio to form complexes and then added to the culture. Cells are incubated for 1 day. Upon internalization and cleavage, positive TAMRA signals are detected by flow cytometry and a normalized internalization index is calculated using IgG1-EN isotype control and anti-CXCR antibody.

[0175] MAPPS assay (MHC-associated peptide proteomics) MAPPS characterizes MHC-II presented peptides on human dendritic cells treated with the test molecule. CD14+ cells isolated from PBMCs of normal human donors are cultured and differentiated into immature DCs by incubation with IL-4 and GM-CSF. On day 4, the medium is replaced with fresh medium containing the test molecule. On day 5, LPS is added to convert the cells into mature DCs. On day 6, the cells are lysed in RIPA buffer containing protease inhibitors. Immunoprecipitation of MHC-II complexes is performed using biotinylated anti-MHC-II antibodies bound to streptavidin beads. Bound complexes are eluted and filtered. Isolated MHC-II peptides are analyzed by mass spectrometry. Peptide identifications are generated by an internal proteomics pipeline using an enzyme-free search algorithm and a bovine / human database with test sequences appended to the database. A KNIME workflow is used to process the identification files from the samples. The peptides identified from the test article are aligned against the parent sequence of the test molecule. The output is used to determine the percentage of donors presenting MHC-II peptides from regions of the test molecule. Compound II, Compound IV, Compound VI, Compound VIII, a compound representative of U.S. Pat. No. 9,023,789 (hereinafter "Compound '789"), and Pramlintide are tested in the MAPPs assay. Compound II, Compound IV, Compound VI, and Compound VIII do not display peptides presented on the MHC-II complex in the assay. Compound '789 displays a peptide cluster spanning residues 8-23 on the MHC-II complex. Pramlintide displays two peptide clusters, both spanning residues 1-34, presented on the MHC-II complex.

[0176] In silico TCEM (T cell exposed motif) analysis This analysis evaluates the potential of a particular peptide cluster identified by MAPPS to activate CD4+ T cells. MAPPS-identified peptide sequences containing non-germline residues are entered into the ImmunoEpitope Database (IEDB) analysis resource MHCII binding prediction page. The IEDB recommended prediction method is selected. For prediction, the 27 most frequent HLA-DR, -DP, and -DQ alleles are considered to cover a significant portion of the population. Each input sequence of 15 residues or more in length is divided into overlapping 15mers (offset of 1 amino acid) across the entire sequence. For each peptide, a percentile rank is generated by comparing the peptide's score to the scores of 5 million random 15mers selected from the SWISSPROT database. Amino acids located at the putative P-1, P2, P3, P5, P7, and P8 positions of the register generate TCEMs and risk is defined based on the presence of non-germline residues at these positions. The possibility of core binding to non-germline residues and multiple alleles is reported in the graphic rendering and taken into account for the immunogenicity risk assessment.

[0177] MS serum binding This assay evaluates off-target binding of test candidates to human serum proteins. Test antibodies are coated onto Immulon 4 HBX microplates. After blocking, human serum is added and incubated overnight. Plates are washed and bound proteins are eluted, reduced, alkylated and digested. Peptides are analyzed by mass spectrometry. Peptide identification is generated by an internal proteomics pipeline using a search algorithm with trypsin enzyme specificity and a human database supplemented with the sequences of the test molecules. Ions are quantified by internal proteomics tools (Chrom-Alignment, Metaconsense, Quant) and analyzed in JMP using the platform One-way analysis / each pair, Student's t-test. Analysis of log2auc of ions using JMP: Approximate Y by X for each ion / Compare means / all pairs, Tukey HSD.

[0178] T cell proliferation assay This assay evaluates the ability of a test antibody or test MAPPS peptide to activate CD4+ T cells by inducing cell proliferation. PBMCs depleted of CD8+ T cells are prepared and labeled with CFSE. Each sample is tested alongside a medium control, keyhole limpet hemocyanin (KLH; a positive clinical benchmark control), a test antibody, or a test MAPPS peptide. Cultures are incubated for 7 days. 。 On day 7, samples are analyzed by flow cytometry.

[0179] Existing reactivity (ACE assay format) This assay evaluates the presence of pre-existing antibodies (ADA) to the test molecule in untreated normal human serum (NHS). Diluted NHS is incubated overnight on Pierce Streptavidin plates coated with biotinylated test molecule. The following day, captured bound proteins are acid eluted, hard coated onto Mesoscale (MSD) plates, and detected with a combination of biotinylated and ruthenium-labeled streptavidin. If anti-drug antibodies are present, they bind to the labeled drug, and the resulting signal is called the Tier 1 signal (expressed as electrochemiluminescence). This signal is confirmed in Tier 2 by adding excess unlabeled test molecule in the detection step, which suppresses the Tier 1 signal. The presence of pre-existing anti-drug antibodies is expressed as the magnitude of the 90th percentile of Tier 2 inhibition. The 90th percentile of Tier 2 inhibition is a statistical tool to assess the magnitude of specificity of Tier 1 reactivity. This 90th percentile is used to rank molecules with respect to ADA risk.

[0180] Table 7. Summary of immunogenicity risk assessment [Table 7] Abbreviations: ACE = acid capture elution; ADA = anti-drug antibody; CDR = complementarity determining region; DC = dendritic cell; H1 = VH CDR1; H2 = VH CDR2; H3 = VH CDR3; L1 = VL CDR1; L2 = VL CDR2; MAPPs = MHC-associated peptide proteomics; MHC = major histocompatibility complex; MS = mass spectrometry; T2 = Tier 2; TCEM = T cell exposed motifs; VH = variable heavy chain; VL = variable light chain; VHFR3 = variable heavy chain framework 3

[0181] Example 17: In vivo efficacy of Compound II in combination with other incretin compounds in Diet Induced Obese (DIO) rats This study is conducted to investigate the effect of Compound II on diabetic and / or obese conditions in DIO rats when administered in combination with other incretin compounds, including a GLP-1 agonist (Compound XVII), an oxyntomodulin analog (Compound XVIII), and triagonists of glucagon, GLP-1, and GIP (Compound XIX). Diet-induced obese (DIO) male Long Evans rats (obtained from Envigo) that have been fed a high-calorie diet (TD95217 (Teklad, Madison, WI)) upon arrival at Lilly are used in the following study. Animals are housed individually in a temperature-controlled (24°C) facility with a 12-hour light / dark cycle (lights on at 22:00) and free access to food (TD95217) and water.

[0182] Randomize the rats according to their body weight so that each experimental group of animals has a similar weight, ranging from 529 to 823 grams.

[0183] Each group contains 5 rats. Vehicle and Compound II (1 nmol / kg) are dissolved in vehicle (20 mM Tris-HCl, pH 8 + 0.02% PS80) and administered by subcutaneous (SC) injection (1 mL / kg) to ad libitum fed DIO rats 30-90 min before the onset of the dark cycle every 3 days for 14 days. SC injections are given on days 1, 4, 7, 10, and 13. Body weight and food intake are measured daily throughout the study. Absolute changes in body weight are calculated by subtracting the body weight of the same animal before the first injection of the molecule.

[0184] At the end of the study, blood is drawn to measure glucose and plasma insulin. Glucose is measured by AccuChek blood glucose meter (Roche, Indianapolis, IN). Insulin is measured by ELISA (MSD, Rockville, MD).

[0185] All data are presented as the mean ± SEM of 5 animals per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's multiple comparison test to compare treatment groups with the vehicle group or with each other. Significant differences were identified at p values ​​less than 0.05.

[0186] Table 8: Effects of Compound II with and without combination with Compound XVII, Compound XVIII, or Compound XIX on body weight and cumulative food intake. [Table 8] * Treatments were administered subcutaneously every 3 days on days 1, 4, 7, 10, and 13. ** Body weight was measured daily and the change in body weight is expressed as the difference in grams from day 1 to day 14. *** Cumulative food intake was the total food consumed over the 14-day treatment period. Statistical analysis was performed by one-way ANOVA followed by Tukey's test. * Compared with the vehicle group, p values ​​were less than 0.05; # p value is less than 0.05 compared with either compound XVII, compound XVIII, or compound XIX group; + Compared with compound II, p value is less than 0.05.

[0187] Combinations of Compound II with Compound XVII, Compound XVIII or Compound XIX result in greater weight loss than Compound II alone.

[0188] SEQ ID NO:1: Compound I γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeN-FGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7.

[0189] SEQ ID NO:2: Compound II γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeN-FGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. The lysine at position 26 is represented by the formula (γE) 2 -CO-(CH 2 ) 18 -CO 2 H.

[0190] SEQ ID NO:3: Compound III γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7.

[0191] SEQ ID NO: 4: Compound IV γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. The lysine at position 26 is represented by the formula (γE) 2 -CO-(CH 2 ) 18 -CO 2 H.

[0192] SEQ ID NO: 5: Compound V KCETATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPILPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7.

[0193] SEQ ID NO:6: Compound VI KCETATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPILPPTEVGSNTY-NH2 There is a thioacetal bridge between the cysteines at positions 2 and 7. The lysine at position 1 has the formula (γE) 2 -CO-(CH 2 ) 18 -CO 2 H.

[0194] SEQ ID NO: 7: Compound VII KCETATCATG-Orn-αMeL-AEFLVRSSHNFGPILPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7.

[0195] SEQ ID NO: 8: Compound VIII KCETATCATG-Orn-αMeL-AEFLVRSSHNFGPILPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. The lysine at position 1 has the formula (γE) 2 -CO-(CH 2 ) 18 -CO 2 H.

[0196] SEQ ID NO: 9: Compound IX γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. Lysine at position 26 is of the formula AEEA 2 -γE-CO-(CH 2 ) 18 -CO 2 H.

[0197] SEQ ID NO: 10: Compound X γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. The lysine at position 26 has the formula γE-AEEA 2 -CO-(CH 2 ) 18 -CO 2 H.

[0198] SEQ ID NO: 11: Compound XI γE-CNTATCATGKLAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. The lysine at position 26 is represented by the formula (γE) 2 -AEEA-CO-(CH 2 ) 18 -CO 2 H.

[0199] SEQ ID NO: 12: Compound XII γE-CNTATCATQ-Orn-LAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. Lysine at position 26 is of the formula AEEA 2 -γE-CO-(CH 2 ) 18 -CO 2 H.

[0200] SEQ ID NO: 13: Compound XIII γE-CGTATCATG-Orn-LAEFLVRSSNNFGPKLPPTEVGSNTY-NH 2 There is a thioacetal bridge between the cysteines at positions 2 and 7. Lysine at position 26 has the formula γE2 -CO-(CH 2 ) 18 -CO 2 H.

[0201] SEQ ID NO: 14: Compound XIV Xaa 1 -C-Xaa 3 -TATCAT-Xaa 10 -Xaa 11 -Xaa 12 -AE-Xaa 15 -LVRSS-Xaa 21 -Xaa 22 -FGP-Xaa 26 -LPPTEVGSNTY-NH 2 (In the sequence, Xaa 1 is K or γE Xaa 3 is E, N, or G Xaa 10 is G or Q, Xaa 11 is Orn or K, Xaa 12 is L or αMeL Xaa 15 is αMeF or F Xaa 21 is N or H Xaa 22 is NMeD, NMeN, or N Xaa 26 is I or K.)

[0202] SEQ ID NO:15: Compound XV; Pramlintide KCNTATCATQRLANFLVHSSNNFGPILPTNVGSNTY-NH 2 There is a disulfide bridge between cysteine ​​at position 2 and cysteine ​​at position 7.

[0203] SEQ ID NO: 16: Compound XVI; hCT CGNLSTCMLGTYTQDFNKFHTFPQTAIGVGAP-NH 2 There is a disulfide bridge between cysteine ​​at position 1 and cysteine ​​at position 7.

[0204] SEQ ID NO:17: hAMY KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY-NH 2 There is a disulfide bridge between cysteine ​​at position 2 and cysteine ​​at position 7.

[0205] SEQ ID NO: 18: Compound XVII H-Aib-EGTFTSDVSSYLEGQAAK ((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) 2 -(γGlu)-CO-(CH 2 ) 16 -CO 2 H)EFIAWLVRGRG

[0206] SEQ ID NO: 19: Compound XVIII H-Aib-QGTFTSDYSKYLDEKKAK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) 2 -(γGlu)-CO-(CH 2 ) 18 -CO 2 H)EFVEWLLEGGPSSG-NH 2

[0207] SEQ ID NO: 20: Compound XIX Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH 2 ) 18 -CO 2 H)AQ-Aib-AFIEYLLEGGPSSGAPPPS-NH 2

[0208] SEQ ID NO:21 YX 1 EGTFTSDYSIX2 LDKIAQKAFVQWLIAGGPSSGAPPPS X 1 is Aib, and X 2 is Aib, and K at the 20th position is (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) 2 -(γGlu) 1 -CO-(CH 2 ) 18 -CO 2 It is chemically modified through conjugation of the K side chain to the epsilon-amino group of H, and the C-terminal amino acid is amidated as a C-terminal primary amide.

[0209] SEQ ID NO:22: Compound I backbone γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeN-FGPKLPPTEVGSNTY-NH 2

[0210] SEQ ID NO:23: Compound III backbone γE-CNTATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPKLPPTEVGSNTY-NH 2

[0211] SEQ ID NO:24: Compound V backbone KCETATCATG-Orn-LAE-αMeF-LVRSSN-NMeD-FGPILPPTEVGSNTY-NH 2

[0212] SEQ ID NO: 25: Compound VII skeleton KCETATCATG-Orn-αMeL-AEFLVRSSHNFGPILPPTEVGSNTY-NH 2

Claims

1. A compound or a pharma- ceutically acceptable salt thereof comprising an array selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:

13.

2. A compound comprising: Xaa1-C-Xaa3-TATCAT-Xaa10-Xaa11-Xaa12-AE-Xaa15-LVRSS-Xaa21-Xaa22-FGP-Xaa26-LPPTEVGSNTY (SEQ ID NO: 14), or a pharma- ceutically acceptable salt thereof, Xaa １ is γE, Xaa ３ is N, Xaa １０ is G, Xaa １１ is Orn, Xaa １２ is L, Xaa １５ is αMeF, Xaa ２１ is N, Xaa ２２ is NMeN, Xaa ２６ is K (SEQ ID NO: 22), or a pharma- ceutically acceptable salt thereof.

3. A compound comprising: Xaa1-C-Xaa3-TATCAT-Xaa10-Xaa11-Xaa12-AE-Xaa15-LVRSS-Xaa21-Xaa22-FGP-Xaa26-LPPTEVGSNTY (SEQ ID NO: 14), or a pharma- ceutically acceptable salt thereof, Xaa １ is γE, Xaa ３ is N, Xaa １０ is G, Xaa １１ is Orn, Xaa １２ is L, Xaa １５ is αMeF, Xaa ２１ is N, Xaa ２２ is NMeD, Xaa ２６ is K (SEQ ID NO: 23), or a pharma- ceutically acceptable salt thereof.

4. Xaa ２６ The lysine in the formula: (γE) ２ -CO-(CH ２ ) １８ -CO ２ 4. The compound of claim 2 or 3, or a pharma- ceutically acceptable salt thereof, attached to a fatty acid linker moiety designated H.

5. A compound comprising: Xaa1-C-Xaa3-TATCAT-Xaa10-Xaa11-Xaa12-AE-Xaa15-LVRSS-Xaa21-Xaa22-FGP-Xaa26-LPPTEVGSNTY (SEQ ID NO: 14), or a pharma- ceutically acceptable salt thereof, Xaa １ is K, Xaa ３ is E, Xaa １０ is G, Xaa １１ is Orn, Xaa １２ is L, Xaa １５ is αMeF, Xaa ２１ is N, Xaa ２２ is NMeD, Xaa ２６ is I (SEQ ID NO: 24), or a pharma- ceutically acceptable salt thereof.

6. A compound comprising: Xaa1-C-Xaa3-TATCAT-Xaa10-Xaa11-Xaa12-AE-Xaa15-LVRSS-Xaa21-Xaa22-FGP-Xaa26-LPPTEVGSNTY (SEQ ID NO: 14), or a pharma- ceutically acceptable salt thereof, Xaa １ is K, Xaa ３ is E, Xaa １０ is G, Xaa １１ is Orn, Xaa １２ is αMeL, Xaa １５ is F, Xaa ２１ is H, Xaa ２２ is N, Xaa ２６ is I (SEQ ID NO: 25), or a pharma- ceutically acceptable salt thereof.

7. Xaa １ The lysine in the formula: (γE) ２ -CO-(CH ２ ) １８ -CO ２ 7. The compound of claim 5 or 6, or a pharma- ceutically acceptable salt thereof, attached to a fatty acid linker moiety designated H.

8. 8. The compound according to claim 2, wherein a thioacetal bridge is present between the cysteine ​​at position 2 and the cysteine ​​at position 7, or a pharma- ceutically acceptable salt thereof.

9. A compound consisting of SEQ ID NO: 2 or a pharma- ceutically acceptable salt thereof.

10. A compound consisting of SEQ ID NO: 4, or a pharma- ceutically acceptable salt thereof.

11. A compound consisting of SEQ ID NO:6, or a pharma- ceutically acceptable salt thereof.

12. A compound consisting of SEQ ID NO:8, or a pharma- ceutically acceptable salt thereof.

13. A compound comprising SEQ ID NO:2, or a pharma- ceutically acceptable salt thereof.

14. A compound comprising SEQ ID NO:4, or a pharma- ceutically acceptable salt thereof.

15. A compound comprising SEQ ID NO:6, or a pharma- ceutically acceptable salt thereof.

16. A compound comprising SEQ ID NO:8, or a pharma- ceutically acceptable salt thereof.

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16, or a pharma- ceutically acceptable salt thereof, and one or more pharma- ceutically acceptable carriers, diluents, or excipients.

18. 17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16, or a pharma- ceutically acceptable salt thereof, for use in the treatment of a condition selected from the group consisting of type 2 diabetes, obesity, dyslipidemia, and NASH.

19. 18. The pharmaceutical composition of claim 17 for use in the treatment of a condition selected from the group consisting of type 2 diabetes, obesity, dyslipidemia, and NASH.

20. 17. Use of a compound according to any one of claims 1 to 16, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a condition selected from the group consisting of type 2 diabetes, obesity, dyslipidemia, and NASH.

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