Amylin receptor agonists

Abstract

The present invention relates to a compound comprising an amylin receptor agonist.The present invention also relates to a pharmaceutical composition comprising such a compound, suitable for oral administration, but not limited thereto.The compound and the pharmaceutical composition comprising it can be used for the medical treatment of subjects with overweight, obesity and related comorbidities.

Description

[Technical Field]

[0001] The present invention relates to a compound that is a dual-acting amylin and calcitonin receptor agonist.The present invention also relates to pharmaceutical compositions that include the above-mentioned compound, including but not limited to, suitable for oral administration.The compound and pharmaceutical compositions that include the compound may be used for the medical treatment of individuals with overweight or obesity (with or without associated comorbidities), diabetes, cardiovascular disease, non-alcoholic steatohepatitis (NASH), and cognitive impairment, such as that caused by Alzheimer's disease. [Background technology]

[0002] Overweight and obesity are abnormal or excessive accumulation of body fat that pose a risk to an individual's overall health. A body mass index (BMI) above 25 is classified as overweight, and a BMI above 30 is classified as obese. Obesity is a major risk factor for numerous serious conditions, including type 2 diabetes and its associated comorbidities, as well as cardiovascular diseases such as heart disease and stroke, which are leading causes of death worldwide. Obesity is now recognized by the World Health Organization (WHO) as an epidemic, even among children. In 2016, 1.9 billion adults worldwide were reported to be obese, and in 2019, 38.3 million children under the age of five worldwide were reported to be obese. According to the WHO, 422 million people worldwide suffer from diabetes, and 1.6 million deaths each year are directly attributable to diabetes. Therefore, there is a significant incentive for individuals and society to prevent and / or treat obesity.

[0003] When diet and exercise alone are not sufficient to reduce the body mass index (BMI) of obese people to an acceptable level, treatment with medications such as liraglutide, semaglutide, orlistat, and naltrexone-bupropion has been shown to cause some weight loss. Nevertheless, bariatric surgery has proven necessary in many cases. Although bariatric surgery is currently the most effective treatment in terms of achieving long-term weight loss, it is an invasive procedure that involves high risks and high costs for patients. Therefore, an effective and minimally invasive treatment would be a significant improvement in the treatment of obesity.

[0004] Amylin is a 37-amino acid polypeptide hormone produced in pancreatic beta (β) cells and secreted simultaneously with insulin. Endogenous amylin has a half-life of approximately 15–20 minutes. It acts primarily through amylin receptors 1–3 (AMYR1–3) to produce its effects in several different organ systems. Amylin inhibits glucagon secretion, delays gastric emptying, conveys satiety, and suppresses appetite; it is an important regulator of energy metabolism in health and disease. Other amylin effects, such as on the cardiovascular system and bone, have also been reported.

[0005] Clinical studies have shown that amylin receptor agonists may be useful in the treatment of overweight, obesity, type 1 diabetes, and / or type 2 diabetes. Currently, one product (Symlin®) containing the amylin receptor agonist (pramlintide acetate) as the active pharmaceutical ingredient is commercially available. Symlin®, a liquid pharmaceutical formulation for subcutaneous administration, is approved for use in patients with type 1 or type 2 diabetes who use basal and prandial insulin and are unable to achieve desirable glycemic control despite optimal insulin therapy. The use of pramlintide in overweight and obese patients has also been investigated in the clinic. Pramlintide has a short biological half-life (less than 1 hour) and requires three doses per day. As a result, there are large diurnal variations in pramlintide plasma levels in patients treated with it.

[0006] A fixed-dose combination of the amylin receptor agonist caglilintide and the GLP-1 receptor agonist semaglutide is currently being investigated for the treatment of overweight and obesity (Non-Patent Document 1). The drug product being investigated is a separate liquid pharmaceutical formulation for subcutaneous use. Clinical trials have demonstrated that the combination of semaglutide and caglilintide induces greater weight loss in obese patients than the maximum approved dose of semaglutide monotherapy, without significantly worsening the side effect profile. Although semaglutide has been successfully formulated for oral administration, caglilintide may not always provide sufficient bioavailability for such an administration route. Therefore, alternative orally available amylin receptor agonists for monotherapy or, but not limited to, combination with semaglutide may prove useful.

[0007] Although current therapeutic options and investigational drugs offer promise, the individuals who are overweight, obese, and / or have associated comorbidities can only hope to be treated with injectable pharmaceutical preparations or medicines with moderate effectiveness at best.There is still a need in the art for more effective medicines that do not cause proportionally increasing side effects and are suitable for oral administration.Therefore, the object of the present invention is to provide an amylin receptor agonist that has suitable bioavailability for oral administration. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Lancet 2021;397:1736-48 Summary of the Invention

[0009] The above-outlined objectives are achieved by the aspects of the present invention, which may also solve further problems that become apparent from the disclosure of the exemplary embodiments.

[0010] In one aspect of the invention, a peptide according to formula I (SEQ ID NO: 36): AX2X3LX5TX7QTX 10 RLAEFLHHX 19 X 20 X 21 X 22 FGX 25 IX 27 X 28 X 29 TX 31 VGX 34 X 35 TX 37 (In the formula, X2 is S or G; X3 is N, H, S, Q, A or E; X5 is A or S; X7 is A or L; X 10 is Q or A, X 19 is S or absent, X 20 is S or absent, X 21 is D, E or absent, X 22 is N, P or absent, X 25 is A, K or P, X 27 is L or P, X 28 is S or P, X 29 is S or P, X 31 is D or E, X 34 is S or P, X 35 is N, D or E, X 37 is Y or P) wherein the peptide comprises a C-terminal amide.

[0011] In one embodiment, the amylin receptor agonist peptide is an amylin analog. Amylin receptor agonists according to Formula I may be considered modified human amylin analogs that lack the disulfide bridge typically found in native human amylin from positions 2 to 7. Thus, in a preferred embodiment, the amylin receptor agonists of the present invention do not contain disulfide bridges. While it has generally been believed that an intact ring structure in amylin is necessary for full biological activity (ACS Pharmacol. Transl. Sci. 2018, 1, 32.49), the inventors surprisingly found that compounds according to Formula I remain potent. Amylin receptor agonists include peptides with alanine (Ala, A) at position 1, leucine (Leu, L) at position 4, and threonine (Thr, T) at position 6. Furthermore, the peptide contains serine (Ser, S) or glycine (Gly, G) at position 2, asparagine (Asn, N), histidine (His, H), serine (Ser, S), glutamine (Gln, Q), alanine (Ala, A), or glutamic acid (Glu, E) at position 3, alanine (Ala, A) or serine (Ser, S) at position 5, and alanine (Ala, A) or leucine (Leu, L) at position 7. Furthermore, the peptide contains glutamine (Gln, Q) at position 8. Without being bound by any particular theory, it is believed that the substitution introduced into the N-terminal sequence of the peptide allows the entire human amylin analog to retain potency while not having an intact ring structure. Thus, a human amylin analog according to Formula I maintains threonine (Thr, T) at position 9, arginine (Arg, R), leucine (Leu, L), and alanine (Ala, A) at positions 11-13, respectively, phenylalanine (Phe, F) and leucine (Leu, L) at positions 15 and 16, respectively, histidine (His, H) at position 18, phenylalanine (Phe, F) and glycine (Gly, G) at positions 23 and 24, respectively, isoleucine (Ile, I) at position 26, threonine (Thr, T) at position 30, valine (Val, V) and glycine (Gly, G) at positions 32 and 33, respectively, and threonine (Thr, T) at position 36.Maintaining complete biological activity without disulfide bridges is believed to be highly advantageous, as it allows for more robust production of amylin receptor agonists.Furthermore, compounds without disulfide bridges are believed to have improved stability in liquid formulations for subcutaneous administration.Thus, in one embodiment, amylin receptor agonists provide improved stability in liquid formulations.Furthermore, the inventors have found that compounds without disulfide bridges can be formulated in a near-neutral pH range, which may facilitate compounding with other compounds that have the same or similar preferred pH range.

[0012] Amylin receptor agonists according to Formula I contain a glutamine at position 8. The inventors have found that amylin receptor agonists containing this substituent are highly potent at both amylin and calcitonin receptors (EC 50 (Values ​​are in the low pM range.) Furthermore, compounds containing the substituents have been shown to be potent in reducing food intake in rats. Thus, in one embodiment, amylin receptor agonists according to Formula I provide improved efficacy in reducing food intake.

[0013] In one embodiment, the amylin receptor agonist is according to Formula I, with the proviso that at least two of the amino acids at positions 21, 31 and 35 are aspartic acid or glutamic acid. The inventors have observed that the amylin receptor agonist in SNAC formulation has high bioavailability in dogs after oral administration. Thus, in one embodiment, the amylin receptor agonist provides improved bioavailability when administered orally.

[0014] In one embodiment, the amylin receptor agonist further comprises an extension moiety attached via an alanine (Ala, A) residue at position 1 or a lysine (Lys, K) residue at position 25. The compounds disclosed herein exhibit a longer half-life compared to their natural ligands. In one embodiment, the amylin receptor agonist has a longer biological half-life relative to the administration interval, thereby reducing the variability of steady-state exposure.

[0015] In one embodiment, amylin receptor agonists may exhibit various properties that make them useful as pharmaceuticals, as described herein.Accordingly, an aspect of the present invention relates to amylin receptor agonists for use as pharmaceuticals.One embodiment relates to amylin receptor agonists for use in treating subjects with an initial body mass index (BMI) of 27 or more (such as 30 or more), optionally in the presence of at least one weight-related comorbidity; optionally in the presence of at least one comorbidity, diabetes; cardiovascular disease; non-alcoholic steatohepatitis; and / or cognitive impairment (such as that caused by Alzheimer's disease).

[0016] A further aspect of the present invention relates to a pharmaceutical composition comprising an amylin receptor agonist and a pharmaceutically acceptable excipient.

[0017] In yet another aspect of the present invention, there is provided a pharmaceutical co-composition comprising an amylin receptor agonist disclosed herein and one or more peptides (such as, but not limited to, insulin peptides) or GLP-1 peptides (such as, but not limited to, semaglutide, liraglutide, or tirzepatide).

[0018] Sequence Listing This application is submitted with an electronic Sequence Listing, the entire contents of which are incorporated herein by reference. SEQ ID NO: 1 represents the amino acid sequence of human amylin (1-37). SEQ ID NOs: 2 to 33 represent the amino acid sequences of peptides in compounds 2 to 43. SEQ ID NO: 34 represents the amino acid sequence of the polypeptide backbone within caglirintide. SEQ ID NO: 35 represents the amino acid sequence of the polypeptide backbone within pramlintide. SEQ ID NO: 36 represents the amino acid sequence of Formula I. [Table 1-1] [Table 1-2] [Table 1-3] DETAILED DESCRIPTION OF THE INVENTION

[0019] Please note that all headings and sub-headings are used herein for convenience only and should not be construed as limiting the invention in any way.

[0020] The use of any and all examples or exemplary language (e.g., "etc.") provided herein is intended merely to better clarify the invention and does not limit the scope of the invention unless otherwise claimed.

[0021] The compounds disclosed herein may be potent amylin receptor agonists with desirable properties, such as being suitable for oral administration. In a first aspect, the present invention provides a peptide according to Formula I (SEQ ID NO: 36): AX2X3LX5TX7QTX 10 RLAEFLHHX 19 X 20 X 21 X 22 FGX 25 IX 27 X 28 X 29 TX 31 VGX 34 X 35 TX 37 , (Wherein, X2 is S or G, X3 is N, H, S, Q, A or E, X5 is A or S, X7 is A or L, and X 10 is Q or A, and X 19 is S or absent, X 20 is S or absent, X 21 is D, E or absent, X 22 is N, P or absent, X 25 is A, K or P, and X 27 is L or P, and X 28 is S or P, and X 29 is S or P, and X 31 is D or E, and X 34 is S or P, and X 35 is N, D or E, and X 37 is Y or P), wherein the peptide comprises a C-terminal amide.

[0022] General definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The practice of the present invention employs, unless otherwise indicated, conventional methods of chemistry, biochemistry, biophysics, molecular biology, cell biology, genetics, immunology, and pharmacology, which are known to those skilled in the art.

[0023] As used herein, the term "peptide" or "peptide sequence" refers to a compound comprising a series of two or more amino acids interconnected through amide (or peptide) bonds. The term peptide is used interchangeably with the terms "polypeptide" and "protein."

[0024] As used herein, the term "analog" generally refers to a polypeptide that has one or more amino acid changes in its sequence compared to a reference amino acid sequence. The amino acid changes may include amino acid additions, deletions, and / or substitutions. Amino acid substitutions, deletions, and / or additions may also be referred to as "mutations." In certain embodiments, an analog "comprises" a specified change. In other specific embodiments, an analog "consists of" or "has" a specified change. When the terms "comprises" or "comprising" are used in reference to amino acid changes in an analog, the analog may have additional amino acid changes compared to its reference sequence. When the terms "consist of" or "having" are used in reference to amino acid changes in an analog, the specified amino acid mutation is the only amino acid change in the analog compared to the reference sequence.

[0025] As used herein, the term "compound" refers to a molecular entity, and therefore, a "compound" may have different structural elements other than the minimum elements defined for each compound or group of compounds. The term compound is used interchangeably with the term "construct." The term "compound" may be used to describe the amylin receptor agonists of the present invention. The compounds of the present invention may be referred to as "compounds," and the term "compound" is also meant to encompass pharmaceutically relevant forms herein, i.e., the present invention also relates to the compounds defined herein, or pharmaceutically acceptable salts, amides, or esters thereof.

[0026] The term "derivative" generally refers to a chemically modified peptide in which one or more substituents or extension moieties are covalently attached to the amino acid sequence of the peptide, for example, via a bond to the alpha position of alanine (Ala, A) at position 1 or the epsilon position of lysine (Lys, K) at position 25.

[0027] amino acid Amino acids are molecules that contain an amine group and a carboxylic acid group, and optionally one or more additional groups, often referred to as side chains. The term "amino acid" includes standard amino acids (genetically encoded) and unnatural amino acids. Non-limiting examples of unnatural amino acids are Aib (α-aminoisobutyric acid) and the d-isomers of standard amino acids. All amino acid residues in polypeptides for which the optical isomer is not specified herein should be understood to mean the I-isomer, unless otherwise specified.

[0028] Receptor agonists A "receptor agonist" or "agonist" is a ligand, such as a compound, that binds to and activates a biological receptor, resulting in a biological response. A full agonist may be defined as an agonist that elicits the same magnitude of response as the natural ligand (see, e.g., "Principles of Biochemistry," A.L. Lehninger, D.L. Nelson, M.M. Cox, Second Edition, Worth Publishers, 1993, page 763). Receptors can be activated either by endogenous agonists, such as endogenous hormones, or by exogenous agonists, such as pharmaceutical agents.

[0029] Amylin receptors Amylin receptor agonists can activate or agonize calcitonin receptors (CTR) and / or amylin receptors (AMYR). Amylin receptors consist of heterodimers of two components: the calcitonin receptor (CTR) and one of three receptor activity-modifying proteins (RAMPs 1-3), resulting in three possible complexes: AMYR1-3. Unless otherwise specified herein, "amylin receptor" refers to at least amylin receptor 3 (AMYR3). Nevertheless, some simultaneous activity on other receptors can be expected.

[0030] Amylin receptor agonists The compounds disclosed herein include amylin receptor agonists. "Amylin receptor agonists" may be defined as a chemical entity that can bind to and activate amylin receptors. In the context of the present invention, "amylin receptor agonists" have the ability to bind to and activate at least AMYR3 complex. Amylin receptor agonists may also have the ability to activate calcitonin receptors and AMYR1-2.

[0031] Examples of endogenous amylin receptor agonists are human amylin and human calcitonin.Human amylin (SEQ ID NO: 1) is a 37 amino acid long polypeptide with an amidated C-terminus and a disulfide bridge between cysteine ​​residues 2 and 7.Both the amidated C-terminus and the disulfide bridge appear to be necessary for the full biological activity of amylin.Examples of exogenous amylin receptor agonists are pramlintide and caglilintide (disclosed in WO2012 / 168432).

[0032] In a preferred embodiment, the amylin receptor agonist according to the present invention is a potent amylin receptor agonist. In one embodiment, the amylin receptor agonist is a potent amylin receptor agonist and a potent calcitonin receptor agonist. The in vitro potency of amylin receptor agonists for the amylin-3 receptor may be measured as described in the assay of Example 4. The potency of a compound is determined by its EC 50 It may be described by a value. 50 represents the concentration of a compound at which 50% of its maximal effect is observed. 50 The lower the value, the more potent the compound. The in vitro potency of amylin receptor agonists for calcitonin receptors may be measured as described in Example 4.

[0033] In one embodiment, when tested as described in the assay of Example 4 for amylin-3 receptor potency, the amylin receptor agonists disclosed herein have an EC50 of less than 300 pM (such as less than 200 pM, such as less than 150 pM), preferably less than 100 pM (such as less than 75 pM), and even more preferably less than 50 pM (such as less than 40 pM, such as less than 30 pM, such as less than 20 pM, such as less than 10 pM, such as less than 7 pM, such as less than 5 pM, such as less than 3 pM, such as less than 2 pM). 50 In certain embodiments, the amylin receptor agonist has an EC value of less than 10 pM as measured by in vitro potency at the amylin-3 receptor according to the assay of Example 4. 50 It has a value.

[0034] In one embodiment, when tested as described in Assay Example 4 for calcitonin receptor potency, the amylin receptor agonists disclosed herein have an EC50 of less than 300 pM (such as less than 200 pM, such as less than 150 pM), preferably less than 100 pM (such as less than 75 pM), preferably less than 50 pM (such as less than 40 pM, such as less than 30 pM, such as less than 20 pM, such as less than 16 pM, such as less than 10 pM, such as less than 7 pM, such as less than 5 pM, such as less than 3 pM, such as less than 2 pM). 50 In certain embodiments, the amylin receptor agonist has an EC value of less than 20 pM as measured by in vitro potency at the calcitonin receptor according to the assay of Example 4. 50 It has a value.

[0035] Human amylin analogues In one embodiment, the amylin receptor agonist peptide is an amylin analog. The amylin receptor agonist disclosed herein is a peptide according to Formula I (SEQ ID NO: 36): AX2X3LX5TX7QTX 10 RLAEFLHHX 19 X 20 X 21 X 22 FGX 25 IX 27 X 28 X 29 TX31 VGX 34 X 35 TX 37 , (Wherein, X2 is S or G, X3 is N, H, S, Q, A or E, X5 is A or S, X7 is A or L, and X 10 is Q or A, and X 19 is S or absent, X 20 is S or absent, X 21 is D, E or absent, X 22 is N, P or absent, X 25 is A, K or P, and X 27 is L or P, and X 28 is S or P, and X 29 is S or P, and X 31 is D or E, and X 34 is S or P, and X 35 is N, D or E, and X 37 is Y or P) and includes peptides comprising a C-terminal amide.

[0036] In one embodiment, the amylin receptor agonist is according to Formula I, wherein X2 is S or G, X3 is H, S, Q or E, X5 is S, X7 is A, and X 10 is Q or A, and X 19 is S or absent, X 20 is S or absent, X 21 is D, E or absent, X 22 is N, P or absent, X 25 is A or P, and X 27 is L or P, and X 28 is S or P, and X 29 is S or P, and X 31 is D and X 34 is S or P, and X 35 is N, D or E, and X 37 is P.

[0037] In one embodiment, the amylin receptor agonist is according to Formula I, wherein X2 is S or G, X3 is H, S, Q or E, X5 is S, X7 is A, and X 10 is Q or A, and X 19 is absent, X 20 is absent, X 21 is absent, X 22 is absent, X 25 is A or P, and X 27 is L or P, and X 28 is S or P, and X 29 is S or P, and X 31 is D and X 34 is S or P, and X 35 is N, D or E, and X 37 is P.

[0038] In one embodiment, the amylin receptor agonist is according to Formula I, wherein X2 is S or G, X3 is H, S or E, X5 is S, X7 is A, and X 10 is Q or A, and X 19 is absent, X 20 X is absent. 21 is absent, X 22 is absent, X 25 is P and X 27 is L and X 28 is P and X 29 is P and X 31 is D and X 34 is S and X 35 is E and X 37 is P.

[0039] In one embodiment, the amylin receptor agonist is according to Formula I, wherein X 27 X 28 X 29 is selected from LPP or PSS.

[0040] In one embodiment, the amylin receptor agonist is according to Formula I, with the proviso that at least two of the amino acids at positions 21, 31, and 35 are aspartic acid (Asp, D) or glutamic acid (Glu, E).

[0041] In one embodiment, the amino acid at position 2 is glycine (Gly, G) or serine (Ser, S).

[0042] In one embodiment, the amino acid at position 3 is glutamic acid (Glu, E), glutamine (Gln, Q), or histidine (His, H).

[0043] In one embodiment, the amino acid at position 5 is serine (Ser, S).

[0044] In one embodiment, the amino acid at position 7 is alanine (Ala, A).

[0045] In one embodiment, the amino acid at position 10 is alanine (Ala, A).

[0046] In one embodiment, the amino acid at position 19 is serine (Ser, S) or absent.

[0047] In one embodiment, the amino acid at position 20 is serine (Ser, S) or absent.

[0048] In one embodiment, the amino acid at position 21 is aspartic acid (Asp, D) or absent.

[0049] In one embodiment, the amino acid at position 22 is asparagine (Asn, N), proline (Pro, P) or absent.

[0050] In one embodiment, the amino acid at position 25 is proline (Pro, P) or alanine (Ala, A).

[0051] In one embodiment, the amino acid at position 27 is proline (Pro, P) or leucine (Leu, L).

[0052] In one embodiment, the amino acid at position 28 is proline (Pro, P) or serine (Ser, S).

[0053] In one embodiment, the amino acid at position 29 is proline (Pro, P) or serine (Ser, S).

[0054] In one embodiment, the amino acid at position 31 is aspartic acid (Asp, D).

[0055] In one embodiment, the amino acid at position 34 is serine (Ser, S) or proline (Pro, P).

[0056] In one embodiment, the amino acid at position 35 is aspartic acid (Asp, D), asparagine (Asn, N), or glutamic acid (Glu, E).

[0057] In one embodiment, the amino acid at position 37 is proline (Pro, P).

[0058] In one aspect, an amylin receptor agonist is provided comprising a peptide selected from the group consisting of SEQ ID NOs: 2 to 33. In certain embodiments, the amylin receptor agonist comprises a peptide selected from any one of the following: AGELSTAQTARLAEFLHHFGPILPPTDVGSETP (SEQ ID NO: 13), ASQLSTAQTARLAEFLHHSSDNFGPILPPTDVGSNTP (SEQ ID NO: 16), ASHLSTAQTARLAEFLHHSSDPFGAIPSSTDVGPDTP (SEQ ID NO: 19).

[0059] In a preferred embodiment, the amylin receptor agonist disclosed herein comprises a C-terminal amide, which is considered to be essential for biological activity.In one embodiment, when the amylin receptor agonist is considered to be a human amylin analog, the proline substituted for tyrosine at the C-terminus comprises an amide group.In a preferred embodiment, the amine group of the C-terminal amide is NH2.

[0060] In one embodiment, the amylin receptor agonist does not contain an intact ring structure. The term "ring structure" refers to any functional group linking two or more amino acid residues together, such as a disulfide bridge. In one embodiment, the amylin receptor agonist does not contain a disulfide bridge. The term "disulfide bridge" with respect to human amylin and its analogs refers to a functional group having the structure RSS-R', also sometimes referred to as an "SS bond."

[0061] In one embodiment, the amylin receptor agonist does not contain any cysteine ​​residues.

[0062] extension part In a preferred embodiment, the amylin receptor agonist comprises an extension moiety, which may also be referred to as a substituent.

[0063] As used herein, the term "extending moiety" refers to a moiety having half-life extending properties, and the extending moiety may be represented by the general formula "AB" or (A)-(B), where (A) is an optional linker and (B) is a protractor. As used herein, the term "protractor" refers to a molecule that has the ability to increase the half-life of the peptide to which it is attached. Thus, the term "extending" refers to half-life extension, and the protractor or extending moiety serves the purpose of extending the half-life of the amylin receptor agonist as disclosed herein.

[0064] As used herein, the term "extension" or "substituent" refers to a moiety that is covalently attached to a peptide. When a substituent is attached to a peptide, the peptide is referred to as being "substituted." When a substituent is covalently attached to a peptide or amino acid residue, the peptide or amino acid is said to "bear" the substituent. A substituent may comprise a series of individually defined moieties, which may be collectively referred to as a "substituent element."

[0065] The prolonged moiety or the substituent has the ability to form a non-covalent bond with albumin, thereby promoting the circulation of the compound in the bloodstream, and thus can have the effect of prolonging the time that the compound remains in the bloodstream, since the aggregates of the fusion compound and albumin slowly disintegrate to release the free form of the compound.Therefore, the substituent or the prolonged moiety as a whole may also be referred to as "albumin binding moiety", and the substituent or the prolonged moiety may be said to have a "prolongation effect".The substituent may include a moiety that is particularly related to albumin binding and thus prolongation, and this moiety may be referred to as a "protractor" or "prolonged moiety".Therefore, the term "substituent" may also be referred to as a "side chain" or "prolonged moiety".

[0066] The extender moiety may include the portion between the protractor (B) and the point of attachment to the polypeptide, which may be referred to as the "linker" (A) or "side chain linker" (A). The linker (A) may include several "linker elements." The linker elements may be selected so that they improve the overall properties of the molecule, for example, so that they improve oral bioavailability, conversion half-life, or prolonged effect, and therefore improve the overall exposure profile of the compound upon oral administration.

[0067] In certain embodiments, the protractor (B) has at least 10 carbon atoms, or at least 15, 20, 25, 30, 35, or at least 40 carbon atoms. In more particular embodiments, the protractor may further comprise at least five heteroatoms, particularly O and N, such as at least 7, 9, 10, 12, 15, 17, or at least 20 heteroatoms (such as at least 1, 2, or 3 N atoms and / or at least 3, 6, 9, 12, or 15 O atoms).

[0068] In certain embodiments, the albumin binding moiety and / or the protracting moiety are lipophilic and / or negatively charged at physiological pH (7.4).

[0069] The albumin binding moiety, protractor, protractor or linker may be covalently attached to the alanine (Ala, A) residue at position 1 or the lysine (Lys, K) residue at position 25 of the peptide by acylation, i.e., via an amide bond formed between the carboxylic acid group (of the albumin binding moiety, protractor, protractor or linker) and the amino group of said alanine (at position alpha) or lysine (at position epsilon), respectively. Additional or alternative conjugation chemistries include alkylation, ester formation, or amide formation, or attachment to cysteine ​​residues, such as by maleimide or haloacetamide (bromo- / fluoro- / iodo- etc.) linkages.

[0070] Unless otherwise specified, when reference is made to the acylation of a lysine residue, it is understood to refer to its epsilon amino group. Unless otherwise specified, when reference is made to the acylation of an alanine residue at position 1, it is understood to refer to its alpha amino group.

[0071] The term "fatty acid" refers to an aliphatic mono- or dicarboxylic acid having 4 to 28 carbon atoms, which is preferably unbranched, and which may be saturated or unsaturated.

[0072] As described above, the extension moiety may be represented by the general formula "AB" or (A)-(B), where (A) is an optional "linker" or "side chain linker" and (B) is a protractor. Each extension moiety is attached to a lysine residue in the peptide backbone of the amylin receptor agonist, such as the alanine residue at position 1 or the lysine residue at position 25. If linker (A) is present, the extension moiety is attached to the polypeptide backbone via linker (A). If linker (A) is absent, (B) is attached to the polypeptide backbone.

[0073] In one embodiment, the extension moiety represented by formula (A)-(B) is covalently attached via a linker (A) to the alpha position of alanine (Ala, A) at position 1 or to the epsilon position of lysine (Lys, K) at position 25 via an amide bond formed between the carboxylic acid group of the extension moiety and the amino group of the alanine (alpha position) or lysine (epsilon position), respectively. In a specific embodiment, the extension moiety is covalently attached to the alpha position (i.e., amino group) of alanine (Ala, A) at position 1. In another specific embodiment, the extension moiety is covalently attached to the epsilon position (i.e., amino group) of lysine (Lys, K) at position 25. In a further embodiment, the amino acid at position 25 is alanine (Ala, A) or proline (Pro, P) when the extension moiety is attached to alanine (Ala, A) at position 1.

[0074] The side chain linker (A) may comprise Ado, Aeep, or Aeeep, sulfonamide, Trx, ε-Lys, Ahx, Glu, γGlu, Gly, Ser, Ala, Thr, and / or a bond.

[0075] The optional side chain linker (A) has the following chemical formula: Formula 1a:*-NH-(CH2)2-(O-(CH2)] k -O-(CH2) n -CO-* Chemical formula 1b: [ka] where k is an integer in the range of 1 to 5 and n is an integer in the range of 1 to 5. When k=1 and n=1, the linker element may be designated Ado, or 8-amino-3,6-dioxaoctanoyl, which may be represented by the following chemical formula: Chemical formula 2a:*NH-(CH2)2-O-(CH2)2-O-CH2-CO-* or Chemical formula 2b: [ka] .

[0076] When k=1 and n=2, the linker element may be designated Aeep, which may be represented by the following chemical formula: Chemical formula 3a: *NH-(CH2)2-O-(CH2)2-O-(CH2)2-CO-* or Chemical formula 3b: [ka] .

[0077] When k=2 and n=2, the linker element may be designated Aeeep, which may be represented by the following chemical formula: Chemical formula 4a:*-NH-(CH2) 2- O-(CH2)2O-(CH2)2-O-(CH2)2-CO-* or Chemical formula 4b: [ka] .

[0078] The optional side chain linker (A) may comprise a sulfonamido-C4 moiety. The sulfonamido-C4 group is a sulfonamido group attached to a 4-butanoyl group and has the following chemical formula: Chemical formula 5a:*NH-S(O)2-CH2-CH2-CH2-CO-* Chemical formula 5b: [ka]

[0079] The optional side chain linker (A) may include Trx, which is also called tranexamic acid, trans-4-(aminomethyl)cyclohexanecarboxylic acid, and has the following chemical formula: Chemical formula 6a:*-NH-CH2-(C6H 10 )-CO-*, or Chemical formula 6b: [ka] .

[0080] The optional side chain linker (A) may comprise epsilon-lysine (ε-Lys).

[0081] The optional side chain linker (A) may include lysine (Lys).

[0082] The optional side chain linker (A) may comprise Ahx, also known as aminocaproic acid, 6-aminohexanoic acid, and defined by: Formula 7a: *-NH-(CH2)5-CO-* or Chemical formula 7b: [ka] .

[0083] In one embodiment, the side chain linker (A) is represented by formula 8a: [ka] or Formula 8b [ka] Contains Glu diradicals such as The Glu diradical may be present p times, where p is an integer ranging from 1 to 3. Formula 8a may also be referred to herein as gamma-Glu, or simply gGlu, due to the fact that it is the gamma-carboxy group of the amino acid glutamic acid used for attachment to another linker element, or to the epsilon-amino group of lysine, or to the alpha group of alanine at position 1. As a result, the amino group of Glu forms an amide bond with the carboxy group of the fatty diacid, or, if present, the gamma-carboxy group of another Glu. Formula 8b may also be referred to as Glu.

[0084] In one embodiment, the linker (A) of the extender moiety comprises a moiety according to Formula 8a. In some embodiments, the linker (A) of the extender moiety comprises one or two moieties according to Formula 8a. In particular embodiments, the linker (A) of the extender moiety consists of one moiety according to Formula 8a. In another particular embodiment, the linker (A) of the extender moiety consists of two moieties according to Formula 8a.

[0085] In one embodiment, the linker (A) of the extender moiety comprises a moiety according to Formula 8b. In some embodiments, the linker (A) of the extender moiety comprises one or two moieties according to Formula 8b. In a particular embodiment, the linker (A) of the extender moiety consists of one moiety according to Formula 8b. In another particular embodiment, the linker (A) of the extender moiety consists of two moieties according to Formula 8b. In one embodiment, Glu is in the L-form.

[0086] The protractor (B) may comprise an acyl group. The acyl group may be branched or unbranched. The acyl group may be saturated or unsaturated. The protractor (B) may comprise a fatty acyl group. The acyl group may be branched or unbranched. The acyl group may be saturated or unsaturated.

[0087] In one embodiment, the protractor (B) comprises a distal carboxylic acid group.

[0088] In one embodiment, the protractor (B) comprises a fatty acid group.

[0089] In one embodiment, the protractor (B) comprises a fatty acid group and an amide group.

[0090] In one embodiment, the protractor (B) comprises a distal carboxylic acid group and an amide group.

[0091] In one embodiment, the protractor (B) comprises an alkyl group.

[0092] In one embodiment, the protractor (B) comprises an aryl group.

[0093] In one embodiment, the protractor (B) comprises a tetrazole group.

[0094] In one embodiment, the protractor (B) comprises a sulfonic acid group.

[0095] In one embodiment, the protractor (B) comprises a phenoxy group.

[0096] In one embodiment, the protractor (B) comprises a benzoic acid group.

[0097] In one embodiment, the protractor (B) is: Chemical formula 9: HOOC-(CH2) n -CO-*, where n is an integer in the range of 8 to 30, which is a C(n+2) diacid, or Chemical formula 10: [ka] (wherein n is an integer in the range of 8 to 30).

[0098] In one embodiment, the protractor (B) of the extender moiety comprises a C14 diacid, a C16 diacid, a C18 diacid, a C20 diacid, a C18-tetrazole, a C14-sulfonic acid, a 4-(9-carboxynonyloxy)benzoic acid, a 4-(10-carboxydecyloxy)benzoic acid, or a 3-(9-carboxynonyloxy)benzoic acid. In certain embodiments, the protractor (B) of the extender moiety comprises a C14 diacid, a C16 diacid, a C18 diacid, or a C20 diacid. Specifically, in one embodiment, the protractor (B) of the extender moiety consists of a C18 diacid. Specifically, in another embodiment, the protractor (B) of the extender moiety consists of a C20 diacid.

[0099] In embodiments in which the extension is attached to the epsilon position of lysine (Lys, K) at position 25, the peptide comprises an N-terminal substituent. In further embodiments, the substituent is covalently attached to the alpha-amino group of the N-terminal amino acid residue, i.e., alanine (Ala, A) at position 1. In further embodiments, the N-terminal substituent is an alkanoyl group or an acyl group. In a further particular embodiment, the N-terminal substituent is an acetyl group. An example of an N-terminal substituted amino acid is Ac-Ala at position 1.

[0100] The compounds disclosed herein have long half-lives compared to their natural ligands. The in vivo half-life of amylin receptor agonists may be evaluated as described in Example 5. The compounds disclosed herein may have long biological half-lives relative to the dosing interval, thereby reducing variability in steady-state exposure. The half-life of an amylin receptor agonist in an animal subject may be as long as about 150 hours or more. In one embodiment, the half-life of an amylin receptor agonist in an animal subject is at least 4 hours. In one embodiment, the half-life of an amylin receptor agonist is greater than 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 hours. In one embodiment, the half-life of an amylin receptor agonist is 15 to 160 hours (e.g., 50 to 155 hours).

[0101] In certain embodiments, the amylin receptor agonist has a half-life [h] of at least 80 in beagle dogs following oral administration, as measured according to the assay of Example 5. More specifically, in one embodiment, the amylin receptor agonist has a half-life [h] of 80 to 150 in beagle dogs following oral administration, as measured according to the assay of Example 5.

[0102] pharmaceutically acceptable salts, amides, or esters The derivatives, analogs, and intermediates of the present invention may be in the form of pharmaceutically acceptable salts, amides, or esters. One aspect of the present invention relates to pharmaceutically acceptable salts of the amylin receptor agonists disclosed herein. Another aspect of the present invention relates to pharmaceutically acceptable esters of the amylin receptor agonists disclosed herein. Yet another aspect of the present invention relates to pharmaceutically acceptable amides of the amylin receptor agonists disclosed herein.

[0103] A salt is formed, for example, by a chemical reaction between a base and an acid, such as: 2NH3 + H2SO4 → (NH4)2SO4.

[0104] Salts may be basic salts, acidic salts, or neither (i.e., neutral salts). Basic salts produce hydroxide ions in water, and acidic salts produce hydronium ions.

[0105] Salts of the derivatives of the invention may be formed between an anionic and cationic group, respectively, with an additional cation or anion, which may be located in the peptide moiety and / or in the side chain of the derivative of the invention.

[0106] Non-limiting examples of anionic groups of the derivatives of the invention include free carboxylic acid groups, if present, in the side chains, as well as in the peptide moiety, which often contains a free carboxylic acid group at the C-terminus and may also contain free carboxylic acid groups in internal acidic amino acid residues such as Asp and Glu.

[0107] Non-limiting examples of cationic groups on a peptide moiety include the free amino group at the N-terminus, if present, and any free amino groups of internal basic amino acid residues such as His, Arg and Lys.

[0108] Esters of the derivatives of the invention may be formed, for example, by reaction of a free carboxylic acid group with an alcohol or phenol, which leads to substitution of at least one hydroxyl group by an alkoxy or aryloxy group.

[0109] Ester formation may involve the free carboxylic acid group at the C-terminus of the peptide and / or any free carboxylic acid group in the side chain.

[0110] Amides of the derivatives of the invention may be formed, for example, by reaction of a free carboxylic acid group with an amine or substituted amine, or by reaction of a free or substituted amino group with a carboxylic acid.

[0111] Amide formation may involve the free carboxylic acid group at the C-terminus of the peptide, any free carboxylic acid group in the side chain, the free amino group at the N-terminus of the peptide, and / or any free or substituted amino group of the peptide within the peptide and / or in the side chain.

[0112] In certain embodiments, peptide or derivative is in the form of pharmaceutically acceptable salt.In another particular embodiment, derivative is in the form of pharmaceutically acceptable amide, preferably amide group is at the C-terminus of peptide.In still further particular embodiments, peptide or derivative is in the form of pharmaceutically acceptable ester.

[0113] compound In one embodiment according to the present invention, the amylin receptor agonist is selected from the group consisting of compounds 2-43.

[0114] In certain embodiments, the amylin receptor agonist is selected from the group consisting of Compound 16, Compound 22, and Compound 25.

[0115] In certain embodiments, the amylin receptor agonist is compound 16 according to the following formula: [ka]

[0116] In certain embodiments, the amylin receptor agonist is compound 22 according to the following formula: [ka]

[0117] In certain embodiments, the amylin receptor agonist is compound 25 according to the following formula: [ka]

[0118] In one embodiment, the amylin receptor agonists disclosed herein reduce food intake in a subject. In one embodiment, administration of the amylin receptor agonists disclosed herein results in a significant reduction in food intake. In one embodiment, the amylin receptor agonists reduce body weight. The in vivo effect of amylin receptor agonists on food intake in rats may be assessed as described in Example 6. In one embodiment, administration of the amylin receptor agonists disclosed herein results in a food intake in the rat of 0-90% (such as 0-80%, for example 0-70%, such as 0-60%, preferably 0-50%, even more preferably 0-40%) compared to vehicle within 0-24 hours after a single subcutaneous injection of 3 nmol / kg or 10 nmol / kg of the co-agonist, where 0% food intake compared to vehicle means that the rat does not eat. In one embodiment, administration of an amylin receptor agonist disclosed herein results in a food intake in rats relative to vehicle of 0-90% (such as 0-80%, for example, 0-70%, such as 0-60%, preferably 0-50%, even more preferably 0-40%) within 24-48 hours after a single subcutaneous injection of 3 nmol / kg or 10 nmol / kg of the co-agonist, where 0% food intake relative to vehicle means that the rat does not eat.

[0119] In one embodiment, the amylin receptor agonist exhibits a reduction in food intake in rats tested according to the assay of Example 6 after administration of 3 nmol / kg. In one embodiment, the amylin receptor agonist exhibits a reduction in food intake in rats tested according to the assay of Example 6, which reduction is visible within 0-24 hours after administration of 3 nmol / kg. In one embodiment, the amylin receptor agonist exhibits a reduction in food intake in rats tested according to the assay of Example 6, which reduction is visible within 0-24 and / or 24-48 hours after administration of 3 nmol / kg. In one embodiment, the amylin receptor agonist exhibits a reduction in food intake of at least 10% (such as at least 15%, 20%, 30%, 40%, 50%, 60%, or 70%) in rats tested according to the assay of Example 6, which reduction is visible within 0-24 hours after administration of 3 nmol / kg. In one embodiment, the amylin receptor agonist exhibits a reduction in food intake of at least 10% (such as at least 15%, 20%, 30%, 40%, 50%, or 60%) in rats tested according to the assay of Example 6, which can be seen within 24-48 hours after administration of 3 nmol / kg. In one embodiment, the amylin receptor agonist exhibits a reduction in food intake of at least 10% (such as at least 15%, 20%, 30%, 40%, 50%, or 60%) in rats tested according to the assay of Example 6, which can be seen within 48-72 hours after administration of 3 nmol / kg.

[0120] Oral bioavailability Oral treatment with pharmacologically active compounds can be hindered by poor bioavailability. The term "bioavailability" refers to the ability of a compound to reach the systemic circulation after administration, and may be quantified as the fractional extent of the compound dose that reaches the systemic circulation upon administration. It is desirable for drugs intended for oral administration to have high oral absorption (i.e., high absorption from the gastrointestinal tract after oral administration), because high absorption can reduce the dose required to reach the intended systemic concentration of the drug, thereby, for example, reducing tablet size and manufacturing costs.

[0121] As used herein, the term "oral bioavailability" refers to the ability of a compound to reach the systemic circulation after oral administration. Oral bioavailability reflects the degree to which a compound is absorbed in the gastrointestinal tract after oral administration. In other words, high oral bioavailability is associated with high oral absorbability. High oral bioavailability of a drug is associated with high drug exposure after oral administration. Oral bioavailability may be measured in beagle dogs with a formulation containing the absorption enhancer N-(8-[2-hydroxybenzoyl]amino) sodium caprylate (SNAC) as described in WO 2019 / 149880.

[0122] Oral bioavailability may be measured as described in Example 5. In one embodiment, a compound of the invention has high oral bioavailability. In one embodiment, a compound of the invention has oral bioavailability similar to that of semaglutide. In one embodiment, a compound of the invention has oral bioavailability comparable to that of semaglutide. In one embodiment, a compound of the invention has oral bioavailability at least as high as that of semaglutide. In one embodiment, a compound of the invention has oral bioavailability suitable for oral administration in humans. In one embodiment, a compound of the invention has oral bioavailability determined in beagle dogs and measured as Cmax / dose [kg / L]. Specifically, in one embodiment, oral bioavailability is determined in beagle dogs measured according to the assay of Example 5. In certain embodiments, oral exposure levels are determined in beagle dogs upon administration of a tablet containing 3 mg of the compound, 300 mg of sodium N-(8-(2-hydroxybenzoyl)amino)caprylate (SNAC), and 7.7 mg of magnesium stearate.

[0123] In one embodiment, the compounds of the invention have an oral bioavailability, measured in beagle dogs as Cmax / dose [kg / L], of at least 0.1, preferably at least 0.15, and most preferably at least 0.20. In one embodiment, the oral bioavailability, measured in beagle dogs as Cmax / dose [kg / L], of between 0.1 and 0.5.

[0124] In one embodiment, the compounds of the invention have oral bioavailability determined in beagle dogs and measured as AUC / dose [kg*hr / L]. In one embodiment, the compounds of the invention have oral bioavailability determined in beagle dogs and measured as AUC / dose [kg*hr / L], where AUC / dose [kg*hr / L] is at least 2, preferably at least 5, and most preferably at least 10. In one embodiment, oral bioavailability is measured in beagle dogs as AUC / dose [kg*hr / L], where AUC / dose [kg*hr / L] is 10-40.

[0125] Pharmaceutical Composition Also disclosed herein are pharmaceutical compositions comprising the amylin analogs disclosed herein. Pharmaceutical compositions comprising an amylin analog or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients may be prepared using methods known to those skilled in the art.

[0126] The term "pharmaceutically acceptable excipient" refers to any ingredient in a pharmaceutical composition that is not the active pharmaceutical ingredient. An excipient may be functional or inert and may serve one or more purposes. For example, an excipient may enhance the absorption of an active substance. An excipient may be, among others, a buffer, antimicrobial preservative, isotonicity agent, carrier, vehicle, filler, binder, lubricant, glidant, disintegrant, flow control agent, crystallization inhibitor, solubilizer, stabilizer, colorant, flavoring agent, surfactant, emulsifier. The amount of each excipient used may vary within ranges conventional in the art.

[0127] The pharmaceutical composition may be suitable for oral administration. Techniques and excipients that may be used to formulate pharmaceutical compositions for oral administration are described in the Handbook of Pharmaceutical Excipients (e.g., 8 thedition, Sheskey et al., Eds., American Pharmaceuticals Association and Pharmaceutical Press, publications department of the Royal Pharmaceutical Society of Great Britain (2017), and later editions), and Remington: The Science and Practice of Pharmacy (e.g., 22nd edition, Remington and Allen, Eds., Pharmaceutical Press (2013), and later editions).

[0128] The pharmaceutical composition may be, for example, a solid pharmaceutical composition (e.g., a compressed tablet or capsule) containing the active pharmaceutical ingredient as a freeze-dried or spray-dried composition, which may be used as is, dissolved before use, or combined with excipients in the composition.

[0129] The pharmaceutical composition comprising the compound of the present invention is suitable for oral administration.In the embodiment for oral administration, the pharmaceutical composition comprising the compound of the present invention is prepared in the form of a tablet, in which the compound is formulated with the absorption enhancer N-(8-[2-hydroxybenzoyl]amino) sodium caprylate (SNAC), as described in, for example, WO 2019 / 149880 or WO 2019 / 215063.Optionally, the compound is further formulated with the lubricant magnesium stearate.

[0130] Alternatively, the pharmaceutical composition may be a liquid formulation, such as an aqueous formulation. Such a liquid composition may be suitable for oral administration or parenteral administration. Liquid compositions suitable for injection can be prepared using conventional techniques in the pharmaceutical industry, involving dissolving and mixing the ingredients as needed to produce the desired final product. Thus, according to a procedure, the compounds described herein are dissolved in a suitable buffer at a suitable pH. The composition may be sterilized, for example, by sterile filtration. Techniques and excipients that may be used to prepare liquid formulations are described in the Handbook of Pharmaceutical Excipients (e.g., 8 th edition, Sheskey et al., Eds., American Pharmaceuticals Association and Pharmaceutical Press, publications department of the Royal Pharmaceutical Society of Great Britain (2017), and later editions), and Remington: The Science and Practice of Pharmacy (e.g., 22 nd edition, Remington and Allen, Eds., Pharmaceutical Press (2013), and subsequent editions).

[0131] In an aspect of the present invention, there is provided a pharmaceutical composition comprising an amylin receptor agonist and / or a pharmaceutically acceptable salt, ester, or amide thereof as disclosed herein.

[0132] In certain embodiments, the pharmaceutical composition is for oral administration. Thus, in one embodiment, the pharmaceutical composition is a solid pharmaceutical composition. More specifically, in one embodiment, the pharmaceutical composition is a tablet.

[0133] In certain embodiments, the pharmaceutical composition is for subcutaneous administration. Thus, in one embodiment, the pharmaceutical composition is a liquid formulation, such as an aqueous formulation. Such a liquid composition may be suitable for oral administration or may be suitable for parenteral administration. Liquid compositions suitable for injection can be prepared using conventional techniques in the pharmaceutical industry, involving dissolving and mixing the ingredients as needed to produce the desired final product. Thus, according to a procedure, the compounds described herein are dissolved in a suitable buffer at a suitable pH. The composition may be sterilized, for example, by sterile filtration. Techniques and excipients that may be used to prepare liquid formulations are described in the Handbook of Pharmaceutical Excipients (e.g., 8 th edition, Sheskey et al., Eds., American Pharmaceuticals Association and Pharmaceutical Press, publications department of the Royal Pharmaceutical Society of Great Britain (2017), and later editions), and Remington: The Science and Practice of Pharmacy (e.g., 22 nd edition, Remington and Allen, Eds., Pharmaceutical Press (2013), and later editions. Preferably, in embodiments where the pharmaceutical composition is in a liquid formulation, the liquid formulation provides improved stability.

[0134] Drug indications Amylin receptor agonists may exhibit various properties that make them useful as pharmaceuticals, as described herein.Therefore, in one embodiment, the amylin analogs disclosed herein or pharmaceutical compositions thereof are used as pharmaceuticals.The amylin analogs disclosed herein may be used for the following medical treatments: (i) prevention and / or treatment of all forms of diabetes, including hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes, MODY (maturity-onset diabetes of the young), gestational diabetes, and / or reduction of HbA1c; (ii) delaying or preventing the progression of diabetic disease, such as the progression of type 2 diabetes, delaying the progression from impaired glucose tolerance (IGT) to insulin-requiring type 2 diabetes, and / or delaying the progression from insulin-nonrequiring type 2 diabetes to insulin-requiring type 2 diabetes; (iii) prevention and / or treatment of eating disorders such as obesity, e.g., reduced food intake, weight loss, appetite suppression, including feelings of satiety; treatment or prevention of binge eating disorder, food cravings, bulimia nervosa and / or obesity induced by the administration of antipsychotics or steroids; reduced gastric motility; and / or delayed gastric emptying, (iv) Weight maintenance after successful weight loss (either drug-induced or diet and exercise-based) - i.e., prevention of weight gain after successful weight loss. (v) prevention and / or treatment of cardiovascular disease, such as delaying or reducing the occurrence of major adverse cardiovascular events (MACE) selected from the group consisting of cardiovascular death, non-fatal myocardial infarction, non-fatal stroke, revascularization, hospitalization for unstable angina, and hospitalization for heart failure; (vi) prevention and / or treatment of nonalcoholic steatohepatitis (NASH); (vii) Prevention and / or treatment of cognitive impairment caused, for example, by Alzheimer's disease.

[0135] In some embodiments, the indication is (i). In some embodiments, the indication is (ii). In still further particular aspects, the indication is (iii). In some embodiments, the indication is (iv). In some embodiments, the indication is (v). In some embodiments, the indication is (vi). In some embodiments, the indication is (vii). In some embodiments, the indication is type 2 diabetes and / or obesity.

[0136] As used herein, the term "treatment" refers to the medical therapy of any human or other vertebrate subject in need thereof. The subject is expected to have undergone a physical examination by a physician or veterinarian who has provided a provisional or definitive diagnosis that will indicate that the use of the particular treatment will be beneficial to the health of the human or other vertebrate. The timing and purpose of the treatment may vary from individual to individual, according to the subject's current health status. Thus, the treatment may be prophylactic (preventative), palliative, symptomatic, and / or curative.

[0137] In some embodiments, the indications are (i) and (iii). In some embodiments, the indications are (ii) and (iii).

[0138] In some embodiments, the subject suffering from obesity is a human, such as an adult human or a pediatric human (including infants, toddlers, and adolescents).

[0139] Body mass index (BMI) is a measure of body fat based on height and weight. The formula is BMI = weight in kilograms / height in meters. 2 A human subject suffering from obesity may have a BMI of ≧30, and the subject may also be referred to as being obese. In some embodiments, a human subject suffering from obesity may have a BMI of ≧35 or a BMI in the range of ≧30 to <40. In some embodiments, the obesity is severe obesity or morbid obesity, in which the human subject may have a BMI of ≧40.

[0140] In some embodiments, the present invention relates to methods for the treatment or prevention of overweight, optionally in the presence of at least one weight-related comorbidity. In some embodiments, the present invention relates to the use of a pharmaceutical composition for the treatment or prevention of overweight, optionally in the presence of at least one weight-related comorbidity. In some embodiments, the overweight subject is a human, such as an adult human or a pediatric human (including infants, toddlers, and adolescents). In some embodiments, the overweight human subject may have a BMI of ≥25 (e.g., a BMI of ≥30, such as a BMI of ≥27, a BMI of ≥35, or a BMI of ≥40). In some embodiments, the overweight human subject has a BMI in the range of 25 to <30 or in the range of 27 to <30. In some embodiments, the weight-related comorbidity is selected from the group consisting of hypertension, diabetes (e.g., type 2 diabetes), dyslipidemia, high cholesterol, and obstructive sleep apnea.

[0141] The term "weight reduction" may include the treatment or prevention of obesity and / or overweight.

[0142] In one embodiment, administration of the compounds disclosed herein is to a patient with a weight-related comorbidity of 30 kg / m or more in the presence of at least one weight-related comorbidity (e.g., hypertension, type 2 diabetes, or dyslipidemia). 2 or over (obese) or 27 kg / m 2 For chronic weight management in adult or pediatric patients with an initial body mass index (BMI) above or equal to (overweight), as an adjunct to a reduced calorie diet and increased physical activity.

[0143] In one embodiment, the co-morbidity is diabetes and / or cardiovascular disease.

[0144] In one embodiment, the amylin receptor agonist is for use in treating a subject with diabetes, such as type II diabetes. In one embodiment, the amylin receptor agonist is for use in treating and / or preventing cardiovascular disease. In one embodiment, the amylin receptor agonist is for use in treating non-alcoholic steatohepatitis (NASH). In one embodiment, the amylin receptor agonist is for use in treating and / or preventing cognitive impairment, for example, cognitive impairment caused by Alzheimer's disease.

[0145] Fixed-dose combination One aspect of the present invention relates to a pharmaceutical combination or combination therapy comprising an amylin receptor agonist disclosed herein and further comprising one or more GLP-1 receptor agonists. In one embodiment, the one or more GLP-1 receptor agonists are peptide GLP-1 receptor agonists or small molecule GLP-1 receptor agonists. In one embodiment, at least one of the one or more GLP-1 receptor agonists is for the treatment and / or prevention of diabetes, cardiovascular disease, NASH, and / or Alzheimer's disease. In one embodiment, the pharmaceutical combination or combination therapy comprises one or more GLP-1-GIP receptor coagonists, wherein the GLP-1-GIP receptor coagonists are peptide GLP-1-GIP receptor coagonists or small molecule GLP-1-GIP receptor coagonists.

[0146] One aspect of the present invention relates to a pharmaceutical combination or combination therapy comprising an amylin receptor agonist disclosed herein and further comprising one or more peptides. In one embodiment, at least one of the one or more peptides is for the treatment and / or prevention of diabetes, cardiovascular disease, NASH, and / or Alzheimer's disease. In a particular embodiment, at least one of the one or more peptides is a GLP-1 peptide. More specifically, in one embodiment, the GLP-1 peptide is a GLP-1 compound, a GLP-1 analog, or a GLP-1 derivative. Even more specifically, in one embodiment, the GLP-1 compound, GLP-1 analog, or GLP-1 derivative is semaglutide or liraglutide.

[0147] In one embodiment, at least one of the one or more peptides is a GLP-1-GIP receptor coagonist.

[0148] In one embodiment, at least one of the one or more peptides is an insulin peptide. More specifically, in one embodiment, the insulin peptide is an insulin compound, an insulin analog, or an insulin derivative. Dosage frequency

[0149] In one embodiment, the amylin analog is formulated for oral administration, such as a tablet. In such an embodiment, the amylin analog disclosed herein may be administered approximately once daily (e.g., once every 12-36 hours, e.g., once every 18-30 hours, e.g., once every 24 hours). The amylin analog disclosed herein may be administered approximately once every other day (e.g., once every 36-60 hours, e.g., once every 42-54 hours, e.g., once every 48 hours). The amylin analog disclosed herein may be administered approximately twice daily (e.g., once every 6-18 hours, e.g., once every 9-15 hours, e.g., once every 12 hours).

[0150] In one embodiment, the amylin analog is formulated for subcutaneous administration, such as in a liquid formulation. In such an embodiment, the amylin analog disclosed herein may be administered approximately once daily (e.g., once every 12-36 hours, such as once every 18-30 hours, such as approximately once every 24 hours). In one embodiment, the amylin analog in a liquid formulation may be administered weekly, biweekly, or monthly.

[0151] Method of production The compounds disclosed herein may be produced, for example, by classical peptide synthesis, e.g., solid phase peptide synthesis using t-Boc or Fmoc chemistry, or other well-established techniques (see, e.g., Greene and Wuts, "Protective Groups in Organic Synthesis", John Wiley & Sons, 1999; Florencio Zaragoza Dorwald, "Organic Synthesis on Solid Phase", Wiley-VCH Verlag GmbH, 2000; and "Fmoc Solid Phase Peptide Synthesis", Edited by W.C. Chan and P.D. White, Oxford University Press, 2000).

[0152] Alternatively, the compounds may be produced by recombinant methods, for example, by culturing host cells containing a DNA sequence encoding the peptide sequence and capable of expressing the peptide in a suitable nutrient medium under conditions that allow expression of the peptide. Non-limiting examples of host cells suitable for expression of these peptides are Escherichia coli, Saccharomyces cerevisiae, and mammalian BHK or CHO cells. Specific examples of methods for preparing the disclosed compounds are included in the Examples.

[0153] A further aspect of the present invention relates to methods for preparing the peptides described herein. In one embodiment, the method for preparing compounds as described herein comprises a step of solid-phase peptide synthesis. Substituents or extensions may be constructed sequentially as part of the solid-phase peptide synthesis or produced separately and attached via an alanine or lysine residue after peptide synthesis.

[0154] Specific Embodiments Aspects of the present invention will now be further described by the following non-limiting embodiments.

[0155] 1. Peptide according to Formula I (SEQ ID NO: 36): AX2X3LX5TX7QTX 10 RLAEFLHHX 19 X 20 X 21 X 22 FGX 25 IX 27 X 28 X 29 TX 31 VGX 34 X 35 TX 37 (In the formula, X2 is S or G; X3 is N, H, S, Q, A or E; X5 is A or S; X7 is A or L; X 10 is Q or A, X 19 is S or absent, X 20 is S or absent, X 21 is D, E or absent, X 22 is N, P or absent, X 25 is A, K or P, X 27 is L or P, X 28 is S or P, X 29 is S or P, X 31 is D or E, X 34 is S or P, X 35 is N, D or E, X 37 is Y or P, An amylin receptor agonist, wherein the peptide comprises a C-terminal amide. 2. The amylin receptor agonist of the preceding embodiment, wherein the peptide is an amylin analog. 3. The amylin receptor agonist of any one of the preceding embodiments, provided that at least two of the amino acids at positions 21, 31, and 35 are aspartic acid (Asp, D) or glutamic acid (Glu, E). 4. Peptide according to formula I (SEQ ID NO: 36): AX2X3LX5TX7QTX 10 RLAEFLHHX 19 X 20 X 21 X 22 FGX 25 IX 27 X 28 X 29 TX 31 VGX 34 X 35 TX 37 (In the formula, X2 is S or G; X3 is H, S, Q or E; X5 is S, X7 is A, X 10 is Q or A, X 19 is S or absent, X 20 is S or absent, X 21 is D, E or absent, X 22 is N, P or absent, X 25 is A or P, X 27 is L or P, X 28 is S or P, X 29 is S or P, X 31 But D, X 34 is S or P, X 35 is N, D or E, X 37is P. 5. Peptide according to formula I (SEQ ID NO: 36): AX2X3LX5TX7QTX 10 RLAEFLHHX 19 X 20 X 21 X 22 FGX 25 IX 27 X 28 X 29 TX 31 VGX 34 X 35 TX 37 (In the formula, X2 is S or G; X3 is H, S, Q or E; X5 is S, X7 is A, X 10 is Q or A, X 19 But he is absent, X 20 But he is absent, X 21 But he is absent, X 22 But he is absent, X 25 is A or P, X 27 is L or P, X 28 is S or P, X 29 is S or P, X 31 But D, X 34 is S or P, X 35 is N, D or E, X 37 is P. 6. Peptide according to formula I (SEQ ID NO: 36): AX2X3LX5TX7QTX 10RLAEFLHHX 19 X 20 X 21 X 22 FGX 25 IX 27 X 28 X 29 TX 31 VGX 34 X 35 TX 37 (In the formula, X2 is S or G; X3 is H, S or E; X5 is S, X7 is A, X 10 is Q or A, X 19 But he is absent, X 20 But he is absent, X 21 But he is absent, X 22 But he is absent, X 25 is P, X 27 But L, X 28 is P, X 29 is P, X 31 But D, X 34 But S, X 35 But E, X 37 is P. 7.X 27 X 28 X 29 2. The amylin receptor agonist of any one of the preceding embodiments, wherein is selected from LPP or PSS. 8. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 2 is glycine (Gly, G) or serine (Ser, S). 9. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 3 is glutamic acid (Glu, E), glutamine (Gln, Q), or histidine (His, H). 10. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 5 is serine (Ser, S). 11. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 7 is alanine (Ala, A). 12. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 10 is alanine (Ala, A). 13. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 19 is serine (Ser, S) or absent. 14. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 20 is serine (Ser, S) or absent. 15. In one embodiment, the amino acid at position 22 is asparagine (Asn, N), proline (Pro, P) or absent. 16. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 25 is proline (Pro, P) or alanine (Ala, A). 17. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 27 is proline (Pro, P) or leucine (Leu, L). 18. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 28 is proline (Pro, P) or serine (Ser, S). 19. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 29 is proline (Pro, P) or serine (Ser, S). 20. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 21 is aspartic acid (Asp, D) or absent. 21. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 31 is aspartic acid (Asp, D). 22. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 34 is serine (Ser, S) or proline (Pro, P). 23. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 35 is aspartic acid (Asp, D), asparagine (Asn, N) or glutamic acid (Glu, E). 24. The amylin receptor agonist of any one of the preceding embodiments, wherein the amino acid at position 37 is proline (Pro, P). 25. An amylin receptor agonist comprising a peptide selected from the group consisting of SEQ ID NOs: 2-33. 26. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 2. 27. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 3. 28. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 4. 29. An amylin receptor agonist, comprising the peptide of SEQ ID NO:5. 30. An amylin receptor agonist, comprising the peptide of SEQ ID NO:6. 31. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 7. 32. An amylin receptor agonist, comprising the peptide of SEQ ID NO:8. 33. Amylin receptor agonists, including peptides of SEQ ID NO: 9. 34. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 10. 35. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 11. 36. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 12. 37. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 13. 38. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 14. 39. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 15. 40. An amylin receptor agonist comprising the peptide of SEQ ID NO: 16. 41. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 17. 42. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 18. 43. Amylin receptor agonists, including peptides of SEQ ID NO: 19. 44. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 20. 45. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 21. 46. ​​An amylin receptor agonist, comprising the peptide of SEQ ID NO: 22. 47. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 23. 48. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 24. 49. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 25. 50. An amylin receptor agonist comprising the peptide of SEQ ID NO: 26. 51. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 27. 52. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 28. 53. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 29. 54. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 30. 55. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 31. 56. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 32. 57. An amylin receptor agonist, comprising the peptide of SEQ ID NO: 33. 58. The following: AGELSTAQTARLAEFLHHFGPILPPTDVGSETP (SEQ ID NO: 13) ASQLSTAQTARLAEFLHHSSDNFGPILPPTDVGSNTP (SEQ ID NO: 16) An amylin receptor agonist comprising a peptide selected from any one of ASHLSTAQTARLAEFLHHSSDPFGAIPSSTDVGPDTP (SEQ ID NO: 19). 59. The amylin receptor agonist of any one of embodiments 25-58, wherein the peptide comprises a C-terminal amide. 60. The amylin receptor agonist of any one of the preceding embodiments, which does not contain an intact ring structure. 61. The amylin receptor agonist according to any one of the preceding embodiments, which does not contain disulfide bridges. 62. The amylin receptor agonist according to any one of the preceding embodiments, wherein the peptide does not contain any cysteine ​​(Cys, C) residues. 63. The amylin receptor agonist of any one of the preceding embodiments, further comprising an extender moiety. 64. The amylin receptor agonist according to the preceding embodiment, wherein the extension moiety is attached to the alpha position of alanine (Ala, A) at position 1 or the epsilon position of lysine (Lys, K) at position 25. 65. The amylin receptor agonist of any one of the preceding embodiments 63 and 64, represented by formula (A)-(B), wherein the extension moiety is attached via a linker (A) to the alpha position of alanine (Ala, A) at position 1 or to the epsilon position of lysine (Lys, K) at position 25. 66. The amylin receptor agonist according to any one of the preceding embodiments 64 and 65, wherein the extension moiety is attached to the alpha position of the alanine (Ala, A) at position 1. 67. The amylin receptor agonist according to any one of the preceding embodiments 63-66, wherein the amino acid at position 25 is alanine (Ala, A) or proline (Pro, P) when the extension moiety is attached to alanine (Ala, A) at position 1. 68. The amylin receptor agonist according to any one of the preceding embodiments 63-67, wherein the extension moiety is attached to the epsilon position of the lysine (Lys, K) at position 25. 69. The amylin receptor agonist according to the preceding embodiment, wherein the N-terminal amino acid is acylated. 70. The amylin receptor agonist according to embodiment 68, wherein the N-terminal amino acid is acetylated. 71. The amylin receptor agonist of embodiment 68, wherein the alanine at position 1 is Ac-Ala. 72. The amylin receptor agonist of any one of the preceding embodiments 65-71, wherein the linker (A) of the extender moiety comprises a moiety according to Formula 8a or Formula 8b. 73. The amylin receptor agonist according to the preceding embodiment, wherein the linker (A) of the extension moiety comprises one or two moieties according to formula 8a or formula 8b. 74. The amylin receptor agonist according to any one of embodiments 72 and 73, wherein the linker (A) of the extension moiety consists of one moiety according to formula 8a or formula 8b. 75. The amylin receptor agonist according to any one of embodiments 72 and 73, wherein the linker (A) of the extension moiety consists of two moieties according to formula 8a or formula 8b. 76. The amylin receptor agonist according to any one of embodiments 72 and 73, wherein the linker (A) of the extension moiety consists of one or two moieties according to formula 8a. 77. The amylin receptor agonist of any one of embodiments 65-76, wherein the protractor (B) of the extender moiety comprises a C14 diacid, a C16 diacid, a C18 diacid, a C20 diacid, a C18-tetrazole, a C14-sulfonic acid, 4-(9-carboxynonyloxy)benzoic acid, 4-(10-carboxydecyloxy)benzoic acid, or 3-(9-carboxynonyloxy)benzoic acid. 78. The amylin receptor agonist according to the preceding embodiment, wherein the protractor (B) of the extension moiety comprises a C14 diacid, a C16 diacid, a C18 diacid, or a C20 diacid. 79. The amylin receptor agonist according to the preceding embodiment, wherein the protractor (B) of the extension moiety consists of a C18 diacid. 80. The amylin receptor agonist according to embodiment 78, wherein the protractor (B) of the extension moiety consists of a C20 diacid. 81. The following formula: [ka] Amylin receptor agonist according to compound 2. 82.The following formula: [ka] Amylin receptor agonist according to compound 3. 83.The following formula: [ka] Amylin receptor agonist according to compound 4. 84.The following formula: [ka] Amylin receptor agonist according to compound 5. 85.The following formula: [ka] Amylin receptor agonist according to compound 6. 86.The following formula: [ka] Amylin receptor agonist according to compound 7. 87.The following formula: [ka] Amylin receptor agonist according to compound 8. 88.The following formula: [ka] Amylin receptor agonist according to compound 9. 89. The following formula: [ka] Amylin receptor agonist according to compound 10. 90.The following formula: [ka] Amylin receptor agonist according to compound 11. 91. The following formula: [ka] Amylin receptor agonist according to compound 12. 92. The following formula: [ka] Amylin receptor agonist according to compound 13. 93. The following formula: [ka] Amylin receptor agonist according to compound 14. 94.The following formula: [ka] Amylin receptor agonist according to compound 15. 95.The following formula: [ka] Amylin receptor agonist according to compound 16. 96. The following formula: [ka] Amylin receptor agonist according to compound 17. 97. The following formula: [ka] Amylin receptor agonist according to compound 18. 98.The following formula: [ka] Amylin receptor agonist according to compound 19. 99.The following formula: [ka] Amylin receptor agonist according to compound 20. 100.The following formula: [ka] Amylin receptor agonist according to compound 21. 101.The following formula: [ka] Amylin receptor agonist according to compound 22. 102.The following formula: [ka] Amylin receptor agonist according to compound 23. 103.The following formula: [ka] Amylin receptor agonist according to compound 24. 104.The following formula: [ka] Amylin receptor agonist according to compound 25. 105.The following formula: [ka] Amylin receptor agonist according to compound 26. 106.The following formula: [ka] Amylin receptor agonist according to compound 27. 107.The following formula: [ka] Amylin receptor agonist according to compound 28. 108.The following formula: [ka] Amylin receptor agonist according to compound 29. 109.The following formula: [ka] Amylin receptor agonist according to compound 30. 110.The following formula: [ka] Amylin receptor agonist according to compound 31. 111.The following formula: [ka] Amylin receptor agonist according to compound 32. 112.The following formula: [ka] Amylin receptor agonist according to compound 33. 113.The following formula: [ka] Amylin receptor agonist according to compound 34. 114.The following formula: [ka] Amylin receptor agonist according to compound 35. 115.The following formula: [ka] Amylin receptor agonist according to compound 36. 116.The following formula: [ka] Amylin receptor agonist according to compound 37. 117.The following formula: [ka] Amylin receptor agonist according to compound 38. 118.The following formula: [ka] Amylin receptor agonist according to compound 39. 119.The following formula: [ka] Amylin receptor agonist according to compound 40. 120.The following formula: [ka] Amylin receptor agonist according to compound 41. 121.The following formula: [ka] Amylin receptor agonist according to compound 42. 122.The following formula: [ka] Amylin receptor agonist according to compound 43. 123. The amylin receptor agonist of any one of the previous embodiments, wherein the peptide comprises a C-terminal amide. 124. The amylin receptor agonist according to the preceding embodiment, wherein the amine group of the C-terminal amide is NH2. 125. An EC of less than 10 pM (such as less than 7 pM, such as less than 5 pM, such as less than 3 pM, such as less than 2 pM) as measured by in vitro potency at the amylin-3 receptor according to the assay of Example 4. 50 2. The amylin receptor agonist of any one of the preceding embodiments, having a value of 126. An EC of less than 20 pM (such as less than 15 pM, such as less than 10 pM, such as less than 5 pM, such as less than 3 pM, such as less than 2 pM) as measured by in vitro potency at the calcitonin receptor according to the assay of Example 4. 50 2. The amylin receptor agonist of any one of the preceding embodiments, having a value of 127. The amylin receptor agonist of any one of the preceding embodiments, having a half-life [h] of at least 80 in beagle dogs following oral administration, as measured according to the assay of Example 5. 128. The amylin receptor agonist of the preceding embodiment, having a half-life [h] of 80 to 150 in beagle dogs following oral administration, as measured according to the assay of Example 5. 129. The amylin receptor agonist according to any one of the preceding embodiments, wherein the oral bioavailability is suitable for oral administration in humans. 130. The amylin receptor agonist of any one of the preceding embodiments, wherein the oral bioavailability is determined in beagle dogs measured according to the assay of Example 5. 131. The amylin receptor agonist of any one of the preceding embodiments, wherein oral bioavailability is determined in beagle dogs upon administration of a tablet containing 3 mg of the compound, 300 mg of sodium N-(8-(2-hydroxybenzoyl)amino)caprylate (SNAC), and 7.7 mg of magnesium stearate. 132. The amylin receptor agonist according to any one of the preceding embodiments, wherein oral bioavailability is measured as Cmax / dose [kg / L]. 133. The amylin receptor agonist according to any one of the preceding embodiments, wherein oral bioavailability is measured as AUC / dose [kg*hr / L]. 134. The amylin receptor agonist of any one of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in Beagle dogs, and wherein Cmax / dose [kg / L] is at least 0.1. 135. The amylin receptor agonist of any one of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in Beagle dogs, and wherein Cmax / dose [kg / L] is at least 0.15. 136. The amylin receptor agonist of any one of the preceding embodiments, wherein the oral bioavailability is measured as Cmax / dose [kg / L] in Beagle dogs, and Cmax / dose [kg / L] is 0.1 to 0.5. 137. The amylin receptor agonist of any one of the preceding embodiments, wherein the oral bioavailability is measured in Beagle dogs as AUC / dose [kg*hr / L], and AUC / dose [kg*hr / L] is at least 10. 138. The amylin receptor agonist of any one of the preceding embodiments, wherein the oral bioavailability is measured in beagle dogs as AUC / dose [kg*hr / L], and AUC / dose [kg*hr / L] is 10-40. 139. The amylin receptor agonist of any one of the preceding embodiments, wherein the amylin receptor agonist reduces appetite. 140. The amylin receptor agonist of any one of the preceding embodiments, wherein the amylin receptor agonist reduces food intake. 141. The amylin receptor agonist of any one of the preceding embodiments, wherein the amylin receptor agonist reduces body weight. 10. The amylin receptor agonist of any one of the preceding embodiments, which exhibits a reduction in food intake in rats tested according to the assay of Example 6 after administration of 142.3 nmol / kg. 10. The amylin receptor agonist of any one of the preceding embodiments, which exhibits a reduction in food intake in rats tested according to the assay of Example 6, which can be seen within 0 to 24 hours after administration of 143.3 nmol / kg. 10. The amylin receptor agonist of any one of the preceding embodiments, which exhibits a reduction in food intake in rats tested according to the assay of Example 6, which can be seen within 0 to 24 and / or 24 to 48 hours after administration of 144.3 nmol / kg. The amylin receptor agonist of any one of the preceding embodiments, which exhibits at least a 10% reduction in food intake in rats tested according to the assay of Example 6, visible within 0 to 24 hours after administration of 145.3 nmol / kg. The amylin receptor agonist of any one of the preceding embodiments, which exhibits a reduction in food intake of at least 10% in rats tested according to the assay of Example 6, visible within 24 to 48 hours after administration of 146.3 nmol / kg. 147. A pharmaceutically acceptable salt of the amylin receptor agonist according to any one of the preceding embodiments. 148. A pharmaceutically acceptable ester of an amylin receptor agonist according to any one of embodiments 1-146. 149. A pharmaceutically acceptable amide of an amylin receptor agonist according to any one of embodiments 1-146. 150. A pharmaceutical composition comprising an amylin receptor agonist and / or a pharmaceutically acceptable salt, ester, or amide thereof according to any one of the preceding embodiments, and one or more pharmaceutically acceptable excipients. 151. The pharmaceutical composition of the preceding embodiment, which is for oral administration. 152. The pharmaceutical composition according to the preceding embodiment, which is a solid pharmaceutical composition. 153. The pharmaceutical composition of the preceding embodiment, which is a tablet. 154. The tablet of any preceding embodiment, wherein the compound is formulated with sodium N-(8-[2-hydroxybenzoyl]amino)caprylate (SNAC). 155. The tablet of the preceding embodiment, wherein the compound is further formulated with the lubricant magnesium stearate. 156. The pharmaceutical composition according to any one of embodiments 153 to 155, for administration approximately once a day (such as once every 12 to 36 hours, such as once every 18 to 30 hours, such as once approximately every 24 hours). 157. The pharmaceutical composition according to embodiment 150, which is a liquid formulation. 158. The pharmaceutical composition of the preceding embodiment, for administration approximately once a week. 159. The pharmaceutical composition according to any one of embodiments 157 and 158, wherein the liquid formulation provides improved stability. 160. An amylin receptor agonist according to any one of embodiments 1 to 146, or a pharmaceutical composition according to any one of embodiments 150 to 159, for use as a medicament. An amylin receptor agonist according to any one of embodiments 1 to 146, or a pharmaceutical composition according to any one of embodiments 150 to 159, for use in treating a subject with an initial body mass index (BMI) of 27 or greater, such as 161.30 or greater. An amylin receptor agonist according to any one of embodiments 1 to 146, or a pharmaceutical composition according to any one of embodiments 150 to 159, for use in treating a subject with an initial body mass index (BMI) of 162.27 or greater and at least one weight-related comorbidity. 163. An amylin receptor agonist according to any one of embodiments 1-146 or a pharmaceutical composition according to any one of embodiments 150-159 for use as an adjunct to a reduced-calorie diet and increased physical activity for chronic weight management in adult subjects with an initial body mass index (BMI) of 30 or greater (obese) or 27 or greater (overweight), in the presence of at least one weight-related comorbidity. 164. The amylin receptor agonist according to any one of embodiments 162 and 163, wherein the comorbidity is diabetes and / or cardiovascular disease. 165. An amylin receptor agonist according to any one of embodiments 1-146 or a pharmaceutical composition according to any one of embodiments 150-159 for use in treating a subject with diabetes, such as type II diabetes. 166. An amylin receptor agonist according to any one of embodiments 1 to 146 or a pharmaceutical composition according to any one of embodiments 150 to 159 for use in the treatment and / or prevention of cardiovascular diseases. 167. An amylin receptor agonist according to any one of embodiments 1-146 or a pharmaceutical composition according to any one of embodiments 150-159 for use in the treatment of non-alcoholic steatohepatitis. 168. An amylin receptor agonist according to any one of embodiments 1 to 146 or a pharmaceutical composition according to any one of embodiments 150 to 159 for use in the treatment and / or prevention of cognitive impairment, such as that caused by Alzheimer's disease. 169. A pharmaceutical combination or combination treatment comprising an amylin receptor agonist according to any one of embodiments 1-146, and further comprising one or more GLP-1 receptor agonists and / or GLP-1-GIP receptor agonists. 170. The pharmaceutical combination or combination treatment of the preceding embodiment, wherein the one or more GLP-1 receptor agonists and / or GLP-1-GIP receptor agonists are peptide GLP-1 receptor agonists or peptide GLP-1-GIP receptor agonists, or small molecule GLP-1 receptor agonists or small molecule GLP-1-GIP receptor agonists. 171. The pharmaceutical combination or combination treatment of any preceding embodiment, wherein the one or more GLP-1-GIP receptor agonists is a peptide GLP-1-GIP receptor agonist or a small molecule GLP-1-GIP receptor agonist. 172. A pharmaceutical combination or combination treatment comprising an amylin receptor agonist according to any one of embodiments 1-146, and further comprising one or more peptides. 173. The pharmaceutical combination or combination treatment according to the preceding embodiment, wherein at least one of the one or more peptides is for the treatment and / or prevention of diabetes, cardiovascular disease, non-alcoholic steatohepatitis, and / or Alzheimer's disease. 174. The pharmaceutical combination or combination treatment according to any one of embodiments 172 and 173, wherein at least one of the one or more peptides is a GLP-1 peptide. 175. The pharmaceutical combination or combination treatment according to the preceding embodiment, wherein the GLP-1 peptide is a GLP-1 compound, a GLP-1 analogue, or a GLP-1 derivative. 176. The pharmaceutical combination or combination treatment according to the preceding embodiment, wherein the GLP-1 compound, GLP-1 analogue or GLP-1 derivative is semaglutide, liraglutide, or tirzepatide. 177. The pharmaceutical combination or combination treatment according to any one of embodiments 169-176, wherein at least one of the one or more peptides is a GLP-1-GIP receptor coagonist. 178. The pharmaceutical combination or combination treatment according to any one of embodiments 169-177, wherein at least one of the one or more peptides is an insulin peptide. 179. The pharmaceutical combination or combination therapy according to the preceding embodiment, wherein the insulin peptide is an insulin compound, an insulin analogue, or an insulin derivative. 180. A method for preparing an amylin receptor agonist according to any one of embodiments 1-146. 181. The method of the preceding embodiment, comprising a step of solid phase peptide synthesis. [Example]

[0156] The following are non-limiting examples for carrying out the present invention.

[0157] Synthesis of amylin analogues and comparative compounds This example provides the identity, materials for making, and methods of synthesis of many compounds according to the invention.

[0158] Also provided are the identities, materials for making, and methods of synthesis of the comparative compounds described herein.

[0159] List of abbreviations The following abbreviations are used below and are in alphabetical order: Ac: Acetyl AUC: Area under the curve Boc: t-butyloxycarbonyl DCM: dichloromethane DIC: diisopropylcarbodiimide DIPEA: N,N-diisopropylethylamine or Hunig's base DMF: dimethylformamide DTT: dithiothreitol EDTA: Ethylenediaminetetraacetic acid ELISA: Enzyme-linked immunosorbent assay Fmoc: 9-fluorenylmethyloxycarbonyl HFIP: 1,1,1,3,3,3-hexafluoro-2-propanol or hexafluoroisopropanol HOBt: 1-hydroxybenzotriazole HPLC: High-performance liquid chromatography Imp: Imidazole propionyl iv: intravenous LCMS or LC-MS: Liquid Chromatography Mass Spectrometry MeCN: acetonitrile Mtt: 4-methyltrityl NHS: N-hydroxysuccinimide NMP: N-methylpyrrolidone Oxyma Pure®: cyano-hydroxyimino-acetic acid ethyl ester PK: Pharmacokinetics QTof: Quantitative time-of-flight method sc: subcutaneous SD: standard deviation SEC-HPLC: Size-exclusion high-performance liquid chromatography SEM: standard error tBu: t-butyl TFA: Trifluoroacetic acid TIPS: Triisopropylsilane Trt: triphenylmethyl or trityl Trx: Tranexamic acid UPLC: Ultra-high performance liquid chromatography

[0160] General method of preparation This section describes general methods for solid-phase peptide synthesis (SPPS), including peptide cleavage from the resin, removal of protecting groups, and purification. LCMS methods for detection and characterization of the resulting peptides are also included.

[0161] Building blocks of fatty acids and special amino acids For the synthesis of octadecanedioic acid mono-tert-butyl ester, see patent application WO 2010 / 102886 (pages 27-28). The corresponding mono-tert-butyl esters of C16 and C20 diacids can be prepared accordingly.

[0162] Fmoc-Gln(Trt)-Thr(ψ Me,Me pro)-OH was commercially available from TechnoComm Ltd.

[0163] Synthesis of the derivatives of the present invention General synthesis method Peptide preparation was carried out using SPPS with Fmoc-based chemistry on a Symphony X from Protein Technologies. The Fmoc-protected amino acids used in this method were the following standard recommendations: Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-His(Trt) ... -lle-OH, Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Bo c)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, Fmoc-Lys(Boc)-OH, Fmoc-Glu-OtBu, and Fmoc-Lys(Mtt)-OH.

[0164] Fmoc-PAL AM resin was used, which was commercially available from NovabioChem. Subsequent amino acids were introduced in a stepwise procedure on a Symphony X peptide synthesizer according to the SPPS principle.

[0165] Fmoc deprotection was achieved with 20% piperidine in DMF containing 0.3 M Oxyma Pure for 2 × 10 min. Introduction of a substituent at the alpha position of the N-terminal amino acid was achieved using standard Fmoc-protected amino acids. Peptide coupling was performed using DIC / Oxyma Pure and collidine. The amino acid / Oxyma Pure solution (0.3 M / 0.3 M in DMF at a 5–10-fold molar excess) was added to the resin first. Then, the same molar equivalent of DIC was added (1.5 M in DMF), followed by collidine (1.5 M in DMF). Most commonly, this was mixed for 2 h. In some cases, the coupling time was increased, additional DIC was added, or the coupling step was repeated. A subsequent capping step was performed using 1 M acetic anhydride and collidine in DMF. The building block Fmoc-Gln(Trt)-Thr(ψ) was synthesized. Me,Me Pro)-OH was introduced at positions corresponding to 8 and 9 of the full-length sequence, where applicable.

[0166] Introduction of the final element of the moiety (i.e., the fatty acid group) was achieved using a suitable building block such as, but not limited to, octadecanedioic acid mono-tert-butyl-ester. Where appropriate, an acetyl group on the N-terminus was introduced by acetylation with 1 M acetic anhydride and collidine in DMF.

[0167] Introduction of a substituent / extension moiety at the epsilon-amino of lysine in the sequence was achieved using Fmoc-Lys(Mtt)-OH. The Mtt group was removed by treatment with HFIP / DCM / TIPS (75:20:5) (5 min) followed by washing with DCM. The resin was then resuspended in HFIP / DCM / TIPS (75:20:5) (2 × 30 min) and subsequently washed with DCM and DMF before the introduction of the substituent / extension moiety at the epsilon-amino of lysine. In these cases, the last amino acid introduced before Mtt removal was Boc-protected, or an acetyl group was introduced as previously described.

[0168] Common cutting methods The peptide was cleaved with TFA / TIPS / HO / DTT (90:4:3:3) for 2 hours, then poured into cold diethyl ether and centrifuged. The ether was decanted, and the peptide was washed twice more with ether.

[0169] General methods for the purification and quantification of derivatives The crude peptide was dissolved in acetic acid / acetonitrile / MQ water (4:2:4) and purified by reverse-phase preparative HPLC (Waters Delta Prep 4000) on a column containing C18 silica gel. Elution was performed with an increasing gradient of MeCN in MQ water containing 0.1% TFA. Relevant fractions were analyzed using UPLC. Fractions containing the pure target peptide were pooled. The resulting solution was analyzed (UPLC, LCMS), and the peptide derivatives were quantified using a CAD-specific HPLC detector (Thermo-Fischer Vanquish HPLC-CAD). The product was dispensed into glass vials. The vials were capped with Millipore glass fiber prefilters. Lyophilization afforded the trifluoroacetate salts of the derivatives as white solids.

[0170] Produced compounds The compounds were prepared using the methods described in the General Methods of Preparation section.

[0171] Example 1: Reference Compounds Compound 1: N 1 Alpha-[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Glu14,Arg17,Pro25,Pro28,Pro29,Gln31,Pro37]-human amylin [ka]

[0172] Example 2: Amylin receptor agonists according to the present invention Compound 2: N 1-[(4S)-4-Carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-human amylin [ka] Compound 3: N 1 -[(4S)-4-Carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-human amylin [ka] Compound 4: N 1 -[(4S)-4-Carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 5: N 1 -[(4S)-4-Carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 6: N 1-[(4S)-4-Carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-human amylin [ka] Compound 7: N 1 -[(4S)-4-Carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Ala1, Gly2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-human amylin [ka] Compound 8: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-human amylin [ka] Compound 9: N 1 -[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-human amylin [ka] Compound 10: N 1-[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Gly2, Ser3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-Human Amylin [ka] Compound 11: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Gly2, Glu3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-Human Amylin [ka] Compound 12: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, Leu4, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-Human Amylin [ka] Compound 13: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Gly2, Ser3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-Human amylin [ka] Compound 14: N 1-[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-human amylin [ka] Compound 15: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Gly2, Ser3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 16: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Gly2, Glu3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 17: N 1 -[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 18: N 1 -acetyl,N{epsilon-25}-[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1,Ser2,Gln3,Leu4,Ser5,Ala7,Gln8,Glu14,His17,Asp21,Lys25,Pro28,Pro29,Asp31,Pro37]-human amylin [ka] Compound 19: N 1 -acetyl,N{epsilon-25}-[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1,Ser2,Gln3,Leu4,Ser5,Ala7,Gln8,Glu14,His17,Asp21,Pro22,Lys25,Pro27,Asp31,Pro34,Asp35,Pro37]-human amylin [ka] Compound 20: N 1 -[(4S)-4-Carboxy-4-(15-carboxypentadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-human amylin [ka] Compound 21: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-human amylin [ka] Compound 22: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, Gln3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-Human amylin [ka] Compound 23: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, Ala3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-Human Amylin [ka] Compound 24: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, Gln3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37-Human amylin [ka] Compound 25: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-human amylin [ka] Compound 26: N1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Pro37]-human amylin [ka] Compound 27: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Glu21, Pro22, Pro27, Glu31, Pro34, Glu35, Pro37]-human amylin [ka] Compound 28: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 29: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, Glu3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 30: N 1-[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]-[Ala1, Gly2, Ser3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 31: N 1 -[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 32: N 1 -[(4S)-4-carboxy-4-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35], des-(19-22)-human amylin [ka] Compound 33: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Leu7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-human amylin [ka] Compound 34: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Leu7, Gln8, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31, Pro37]-human amylin [ka] Compound 35: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Leu7, Gln8, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka] Compound 36: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35]-Human amylin [ka] Compound 37: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35]-Human amylin [ka] Compound 38: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro25, Pro28, Pro29, Asp31]-human amylin [ka] Compound 39: N 1 -[(4S)-4-Carboxy-4-(19-carboxynonadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Asp21, Pro22, Pro27, Asp31, Pro34, Asp35, Pro37]-human amylin [ka] Compound 40: N 1 -acetyl,N{epsilon-25}-[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1,Ser2,His3,Leu4,Ser5,Ala7,Gln8,Ala10,Glu14,His17,Asp21,Pro22,Lys25,Pro27,Asp31,Pro34,Asp35,Pro37]-human amylin [ka] Compound 41: N 1 -acetyl,N{epsilon-25}-[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1,Ser2,His3,Leu4,Ser5,Ala7,Gln8,Ala10,Glu14,His17,Asp21,Lys25,Pro28,Pro29,Asp31,Pro37]-human amylin [ka] Compound 42: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35], des-(19-22)-human amylin [ka] Compound 43: N 1 -[(4S)-4-Carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]-[Ala1, Ser2, His3, Leu4, Ser5, Ala7, Gln8, Ala10, Glu14, His17, Pro25, Pro28, Pro29, Asp31, Glu35, Pro37], des-(19-22)-human amylin [ka]

[0173] Example 3: LCMS characterization of synthesized compounds LCMS analysis was performed on a setup consisting of a Waters Acquity UPLC H Class system and a Waters Xevo G2-xS QTof. Eluents: A: MQ water, B: MeCN, C: 2% formic acid + 0.1% TFA in MQ water.

[0174] The analysis was performed at room temperature (column temperature 60 °C) by injecting an appropriate amount of sample onto the column, which was eluted with a gradient of A, B, and 5% C. The UPLC conditions, detector settings, and mass spectrometer settings were as follows: Column: Waters Acquity BEH, C-18, 1.7 μm, 2.1 mm x 50 mm. Gradient: 4.0 min, linear 5% to 95% B at 0.4 ml / min. Total run time: 7.0 min. Detection: MS resolution mode, ionization method: ES. Scan: 50 to 5000 amu.

[0175] The monoisotopic masses of the examples were recorded and the observed and calculated values ​​are shown in Table 1. [Table 2-1] [Table 2-2] [Table 2-3]

[0176] Example 4: In vitro potency assays of human amylin receptors and human calcitonin receptors Amylin receptor assay To determine the ability of a compound to activate or agonize the amylin receptor, an in vitro potency assay on cells expressing the human amylin receptor (hAMYR3) can be performed as follows.

[0177] Assay principle Activation of hAMYR3 leads to an increase in the cellular concentration of cAMP. As a result, transcription is activated by promoters containing multiple copies of the cAMP response element (CRE). Therefore, it is possible to measure hAMYR3 activity using a CRE-luciferase reporter gene introduced into baby hamster kidney (BHK) cells co-expressing hAMYR3.

[0178] Cells and assay reagents The BHK cell line was stably transfected with the human calcitonin receptor(a) and a CRE-responsive luciferase (CRE-Luc) reporter gene according to methods known to those skilled in the art (Hollex-1 cell line, obtained from Zymogenetics as described in U.S. Pat. No. 5,622,839). The cell line was further transfected with human receptor-modifying protein 3 (RAMP3) using standard methods, which converts the human calcitonin receptor to the human amylin-3(a) receptor.

[0179] Cell stocks were prepared by culturing the stably transfected BHK hAMYR3 / CRE-Luc cell line in growth medium consisting of DMEM (Gibco, 31966-021) supplemented with 10% FBS (Gibco, 16140-071), 1% penicillin / streptomycin (Gibco, 15140-122), 0.5 mg / mL geneticin (Gibco, 10131-027), 0.4 mg / mL hygromycin (Invitrogen, 10687010), and 250 nM methotrexate (Sigma, A6770). Cells at approximately 80-90% confluence were washed once with PBS (Gibco 14190-094) and detached from the cell flask using Versene (Gibco, 15040-033) or TrypLE™ (Gibco, 12605-010). After centrifugation, the cell pellet was lysed and diluted to approximately 2.5 x 10E6 cells / mL in Recovery™ Cell Culture Freezing Medium (Gibco, 12648-010). Cells were aliquoted and stored at -180°C until use.

[0180] The assay buffer consisted of DMEM without phenol red (Gibco, 11880-028) and supplemented with 1X GlutaMAX (Gibco, 35050-038), 10 mM HEPES (Gibco, 15630-056), and 1% (w / v) ovalbumin (Sigma, A5503).

[0181] procedure To perform the assay, BHK hAMYR3 / CRE-Luc cells were thawed, washed once with PBS, and seeded at a cell density of 4,000 cells / well in 40 μL of growth medium in white 384-well culture plates (PerkinElmer, 6007688) the day before the experiment. Plates were incubated overnight at 37°C in 5% CO2. On the day of the assay, cells were washed once in assay buffer. Serial dilutions of comparison compounds and amylin analogs (7-fold dilutions, 7 concentrations per compound, with one well containing assay buffer only) were performed in assay buffer, often starting at approximately 10 nM in a 96-well plate. 30 μL of each concentration was added to the 384-well assay plate containing the cells. The assay plate was incubated at 37°C in 5% CO2 for 3 hours, after which 30 μL of SteadyLite Plus (PerkinElmer, 6066759) was added to each well. The assay plate was sealed and incubated at room temperature with gentle shaking for 5 minutes, followed by 30 minutes without shaking, protected from light. Luminescence was detected with a luminescence plate reader, e.g., Synergy 2 (BioTek). EC 50 Values ​​[pM] were calculated by nonlinear curve fitting applying a four-parameter logistic model (Hill slope = 1.5, shared bottom response within each plate) using GraphPad Prism or by TIBCO Enterprise Runtime for R (TIBCO Software, Palo Alto, CA, USA).

[0182] Calcitonin receptor assay To determine the ability of a compound to activate or agonize the calcitonin receptor, an in vitro potency assay on cells expressing the human calcitonin receptor (hCTR) can be performed as follows.

[0183] Assay principle Activation of hCTR leads to an increase in the cellular concentration of cAMP. Consequently, transcription is activated by promoters containing multiple copies of the cAMP response element (CRE). Therefore, hCTR activity can be measured using a CRE-luciferase reporter gene introduced into baby hamster kidney (BHK) cells co-expressing hCTR.

[0184] Cells and assay reagents The BHK cell line was stably transfected with the human calcitonin receptor (a) and a CRE-responsive luciferase (CRE-Luc) reporter gene according to methods known to those skilled in the art (Hollex-1 cell line, obtained from Zymogenetics as described in U.S. Pat. No. 5,622,839).

[0185] Cell stocks were prepared by culturing stably transfected BHK hCTR / CRE-Luc cell lines in growth medium consisting of DMEM (Gibco, 31966-021) supplemented with 10% FBS (Gibco, 16140-071), 1% penicillin / streptomycin (Gibco, 15140-122), 0.5 mg / mL Geneticin (Gibco, 10131-027), and 250 nM methotrexate (Sigma, A6770). Cells at approximately 80-90% confluence were washed once with PBS (Gibco, 14190-094) and detached from the cell flask using Versene (Gibco, 15040-033) or TrypLE™ (Gibco, 12605-010). After centrifugation, the cell pellet was lysed and diluted to approximately 2.5 x 10E6 cells / mL with Recovery™ Cell Culture Freezing Medium (Gibco, 12648-010). Cells were aliquoted and stored at -180°C until use.

[0186] The assay buffer consisted of DMEM without phenol red (Gibco, 11880-028) and supplemented with 1X GlutaMAX (Gibco, 35050-038), 10 mM HEPES (Gibco, 15630-056), and 1% (w / v) ovalbumin (Sigma, A5503).

[0187] procedure To perform the assay, BHK hCTR / CRE-Luc cells were thawed, washed once with PBS, and seeded at a cell density of 4,000 cells / well in 40 μL growth medium in white 384-well culture plates (PerkinElmer, 6007688) the day before the experiment. Plates were incubated overnight at 37°C in 5% CO2. On the day of the assay, cells were washed once in assay buffer. Serial dilutions of comparison compounds and amylin analogs (7-fold dilutions, 7 concentrations per compound, with one well containing assay buffer only) were performed in assay buffer, often starting at approximately 10 nM in a 96-well plate. 30 μL of each concentration was added to the 384-well assay plate containing the cells. The assay plate was incubated at 37°C in 5% CO2 for 3 hours, after which 30 μL of SteadyLite Plus (PerkinElmer, 6066759) was added to each well. The assay plate was sealed and incubated at room temperature with gentle shaking for 5 minutes, followed by 30 minutes without shaking, protected from light. Luminescence was detected with a luminescence plate reader, e.g., Synergy 2 (BioTek). EC 50 Values ​​[pM] were calculated by nonlinear curve fitting applying a four-parameter logistic model (Hill slope = 1.5, shared bottom response within each plate) using GraphPad Prism or by TIBCO Enterprise Runtime for R (TIBCO Software, Palo Alto, CA, USA). [Table 3] [Table 4]

[0188] The data in Table 2 show that all compounds tested agonized both human AmyR3 and CalcR, and most had EC values ​​comparable to that of caglintide. 50 The values ​​(Table 3) indicate that it agonizes both receptors.

[0189] The compounds listed in Table 2 are all compounds according to the present invention.

[0190] Example 5: Oral Pharmacokinetic Profile in Beagle Dogs Preparation of tablets for pharmacokinetic studies in beagles To be able to assess oral exposure after tablet administration, tablet compositions containing the test substance and SNAC (sodium N-(8-(2-hydroxybenzoyl)amino)caprylate) were prepared by mixing the test substance with roller-compacted SNAC and magnesium stearate, e.g., as described in WO 2019 / 149880.

[0191] A method for measuring oral PK profiles in beagle dogs This assay was performed to measure the oral bioavailability of compounds. The assay determined the exposure of test compounds after oral administration in beagle dogs, as described by relevant pharmacokinetic parameters and plasma concentration curves.

[0192] Preparation of tablets for oral administration: The tablets containing the test compound used for the assays described herein were immediate-release SNAC-based tablets. The test compound was spray-dried as a neutral sodium salt (pH 7-8). Dry granulation was performed by roller compaction on a Gerteis MINI-PACTOR. Tablets containing 3 mg of test compound, 300 mg of sodium N-(8-(2-hydroxybenzoyl)amino)caprylate (SNAC), and 7.7 mg of magnesium stearate were produced on a Kilian Style One using a 7.2 x 12 mm punch.

[0193] Determination of absorption after oral administration: Eight male beagle dogs, weighing approximately 10-15 kg at the start of the study and aged 1-8 years, were used. The dogs were group-housed in enclosures (12-hour light:12-hour dark) and individually and restrictively fed Royal Canin Medium Adult dogs (Royal Canin Products, China Branch, or Brogaarden A / S, Denmark) once daily. Exercise and group socialization were permitted daily whenever possible. Dogs were used for repeated pharmacokinetic studies, with an appropriate washout period between successive doses. An appropriate acclimation period was allowed before the start of the first pharmacokinetic study. All animal handling, dosing, and blood sampling were performed by trained and experienced staff. The dogs were fasted overnight before the study and again for 0-4 hours after dosing. Furthermore, access to water was restricted from 1 hour before dosing until 4 hours after dosing, but dogs had free access to water throughout the study.

[0194] Tablets containing test compounds were administered in the following manner: 10 minutes before tablet administration, dogs were subcutaneously administered approximately 3.2 nmol / kg of glucagon. The tablet was placed at the back of the dog's mouth to prevent chewing. After that, the mouth was closed, and 10 mL of tap water was given by syringe to facilitate swallowing of the tablet. In order to adequately cover the complete plasma concentration-time absorption profile of the compound, blood was collected at predetermined time points up to 288 hours after administration. For each blood collection time point, approximately 1.2 mL of whole blood was collected into a 1.5 mL EDTA-coated tube, and the tube was gently rotated to mix the sample with EDTA. The blood sample was then kept on ice until centrifugation (4 minutes, 4 ° C, 4000 rpm). Plasma was pipetted into a microtube on dry ice and kept at -20 ° C until analysis. Blood samples were collected as needed, for example, from a Venflon in the cephalic vein of the front leg for the first 2 hours, and then from the jugular vein for the remaining time points using a syringe (the first few drops were allowed to drain from the Venflon during the sample to avoid heparinized saline from the Venflon).

[0195] Bioanalysis was performed as follows: Plasma concentrations of test compounds were assayed by plasma protein precipitation and analyzed by liquid chromatography-mass spectrometry (LC-MS). Calibrators were prepared by spiking blank dog plasma with the analyte to reach final concentrations typically ranging from 0.5 to 200 nM. Calibrators, plasma blanks, or study samples were prepared for LC-MS by protein precipitation by adding 3 volumes of ethanol, followed by centrifugation at 4000 rpm for 1 hour at 4°C. The supernatant was diluted with 2 volumes of Milli-Q water containing 0.1% formic acid before injection into the LC-MS system. The system used was a Transcend II Interface Module SRD3200 system from Thermo Scientific (Waltham, MA, USA) coupled to an Orbitrap Exploris 240 mass spectrometer from Thermo Scientific. The LC was equipped with a TurboFlow Cyclone Column 0.5 × 50 mm (CH-953288, Thermo Scientific) as the first-dimension trapping column and a Poroshell 120 SB-C18 2.7 μm (2.1 × 50 mm, Agilent, Santa Clara, CA, USA) as the analytical column. The mobile phase composition of the loading pump was as follows: Mobile phase A consisted of 95% milli-Q water, 2.5% acetonitrile, 2.5% methanol, and 0.1% formic acid; Mobile phase B consisted of 47.5% acetonitrile, 47.5% methanol, 5% milli-Q water, and 0.1% formic acid. The target analytes were loaded from the Turboflow column to the second-dimension analytical column at 30% B. Gradient elution was performed with an elution pump using mobile phase A (95% milli-Q water, 2.5% acetonitrile, 2.5% methanol, and 0.1% formic acid) and mobile phase B (47.5% acetonitrile, 47.5% methanol, 5% milli-Q water, and 0.1% formic acid), with a gradient from 0% mobile phase B to 60% mobile phase B in 0.25 min, from 60% mobile phase B to 80% mobile phase B in 1.17 min, and then from 80% mobile phase B to 100% mobile phase B in 1.17 min.The Orbitrap Exploris 240 was operated in positive ionization mode with parallel reaction monitoring (PRM) scan mode. A linear calibration curve (1 / ×2 weighting) was used to calculate test compound concentrations in plasma samples and determine maximum plasma concentrations (Cmax). Quality control samples were included for the analytes. Deviations between nominal and calculated concentrations in calibrators and quality control samples were less than 15%, and LLOQ samples were less than 20%. Plasma concentration (vs. time) profiles of test compounds were assessed, and standard pharmacokinetic parameters were estimated by noncompartmental analysis (NCA) using WinNonlin Phoenix 64 (version 8.3.3, CERTARA). Results were reported as dose-corrected maximum plasma concentration (Cmax / dose), dose-corrected area under the curve (AUC / dose), and half-life (T1 / 2).

[0196] The AUC from the time of administration was extrapolated to infinity based on the final observed concentration (_obs) or final predicted concentration (_pred). The "Linear Ascending Log Descending" option was selected.

number

number

number

[0197] Using WinNonlin NCA analysis, the first-order rate constant (Lambda-Z) was estimated by linear regression of time versus log concentration. The first-order rate constant was associated with the terminal (log-linear) portion of the curve. The software calculated the Lambda-Z value and provided the half-life (T) according to the following equation:

number

[0198] All compounds in Table 4 demonstrated plasma exposure following oral administration of SNAC (Sodium N-(8-(2-hydroxybenzoyl)amino)caprylate) tablets containing magnesium stearate to beagle dogs.

[0199] The compounds in Table 4 showed improved exposure levels compared to the proximal caglilintide analog Compound 1 (Table 5). Furthermore, the compounds in Table 4 showed long T 1 / 2 It presents.

[0200] The compounds listed in Table 4 are all compounds according to the present invention.

[0201] Example 6: Food intake study in a rat model Preparation of formulation for subcutaneous administration to rats The lyophilized compound was dissolved in vehicle, 8 mM phosphate, 250 mM glycerol, 0.007% polysorbate 20, and the pH was adjusted to 7.4 with 1 N NaOH.

[0202] The final formulation was sterile filtered and filled into penfill cartridges for administration.

[0203] Experimental Protocol for Testing Appetite Efficacy Using an Ad libitum-Fed Rat Model Normal weight Sprague Dawley (SD) rats from Taconic Europe, Denmark were used in acute food intake studies. The purpose of these studies was to examine the in vivo effects on food intake and to provide an indication of the duration of action of the compounds.

[0204] Experiments were performed with permission from the Danish Animal Experiments Inspectorate and in accordance with the Danish Act governing animal experimentation (1987) and the guidelines of the National Institutes of Health (Publication no. 85-23) and the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes (Council of Europe no. 123, Strasbourg 1985).

[0205] Prior to the start of the study, rats were allowed to acclimate to the experimental setup for 14 days. During this period, animals were handled at least twice. Immediately after arrival, rats were placed under a reverse light cycle (10 AM to 10 PM or 11 AM to 11 PM dark) and transferred to one of three food intake measurement systems: FeedWin, BioDaq, or HM2. Rats were housed singly in the FeedWin and BioDaq systems, and three per cage in the HM2 system (rats were tagged with ID chips). During the acclimation period, during which rats became accustomed to the new light cycle and diet, animals had free access to food and water. Because rats are nocturnal animals (e.g., they are most active and consume the majority of their calories during the dark period), they were administered in the morning, just before falling asleep, to prevent interference with their natural feeding patterns, thereby minimizing data variability and improving sensitivity. Each amylin analog was tested in groups of five to six rats. A vehicle group of 5-6 rats was included in each experiment and used as a reference point. For studies in the HM2 system, rats in the same cage received different treatments to eliminate potential interference from technical variations between cages. At the start of treatment, rats received a single subcutaneous dose of the extended amylin analog at a dose of 3 nmol / kg, based on their body weight. The time of administration was recorded for each rat.

[0206] After administration, rats were returned to their home cages, where they had access to food and water.Food consumption was continuously recorded individually by online registration up to 72 hours after administration.At the end of the experiment, body weight was measured again, and animals were euthanized by CO2 overdose.

[0207] Data from the food intake study in rats are shown in Table 6. [Table 7] [Table 8]

[0208] The amylin comparator compound caglilintide (Table 7) comprises a peptide according to SEQ ID NO: 34 and is further disclosed in WO 2012 / 168432.

[0209] As can be inferred from the data presented in Table 6, after administration of amylin analogs to rats, many of them were observed to induce significant inhibition of food intake compared to vehicle treatment. Overall, the compounds in Table 6 showed improved reduction in food intake compared to the comparator caglilintide (Table 7).

[0210] The compounds listed in Table 6 are all compounds according to the present invention.

[0211] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those skilled in the art. It is therefore to be understood that the appended claims are intended to cover all such modifications and changes that fall within the true spirit of the invention.

Claims

1. The following formula: 【Chemistry 16】 compound 16, Amyrin receptor agonists, or pharmaceutically acceptable salts, esters, or amides thereof.

2. A pharmaceutical composition comprising the amyrin receptor agonist described in claim 1, and / or a pharmaceutically acceptable salt, ester, or amide thereof, and one or more pharmaceutically acceptable excipients.

3. An amyrin receptor agonist according to claim 1, or a pharmaceutical composition according to claim 2, for use as a pharmaceutical.

4. an amyrin receptor agonist according to claim 1, or a pharmaceutical composition according to claim 2, for use in the treatment of subjects having an initial body mass index (BMI) of 27 or more, for example 30 or more, in which at least one weight-related comorbidity is present; diabetes mellitus; cardiovascular disease; non-alcoholic steatohepatitis, and / or cognitive impairment, for example cognitive impairment caused by Alzheimer's disease.

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