Ham15-52 analogues with improved amylin receptor (hamy3r) potency

Description

FIELD OF THE INVENTION

[0001] The present invention relates to hAM 15-52 analogues with improved amylin receptor (hAMY3R) potency (hAMY3R-EC 50 ≤ 250 pM) and which are largely based on the sequence of the human adrenomedullin fragment hAM 15-52 . The invention further relates to hAM 15-52 analogues that are selective amylin receptor (hAMY3R) agonists (hAMY3R-EC 50 ≤ 250 pM and an hAM1R-EC 50 ≥ 25 nM). In particularly, the present invention is based on the realization that the human adrenomedullin fragment hAM 15-52 may be converted into highly selective amylin receptor agonists (hAMY3R-EC 50 ≤ 250 pM and an hAM1R-EC 50 ≥ 25 nM) by replacing the amino acid in position X 11 in hAM 15-52 together with one or more of the amino acids in the positions X 4 , X 37 and / or X 38 in hAM 15-52 . Thus, replacing at least two amino acids on hAM 15-52 resulted in hAM 15-52 analogues with AMY3R and AMIR potencies comparable to hAMY 1-37 (amylin). The invention further relates to pharmaceutical compositions comprising such hAM 15-52 analogues and their medical use in the treatment of a medical condition, such as obesity and / or diabetes.BACKGROUND OF THE INVENTION

[0002] Obesity is a medical condition in which excess body fat has accumulated to the extent that it has a negative impact on health. It is affecting a huge number of individuals worldwide and increasing rapidly in certain parts of the world. The World Health Organisation (WHO) estimated that in 2016, approximately 650 million people were obese worldwide. Obesity is defined as a body mass index (BMI) above 30. Obesity is considered a major risk factor for developing a variety of medical conditions, such as cardiovascular diseases (e.g. hypertension, atherosclerosis, heart attacks or stroke), NASH musculoskeletal disorders, certain kinds of cancers, depression and diabetes type II, and hence is detrimental to human health. Cardiovascular diseases and diabetes are two main diseases associated with obesity. A large amount of research has been conducted in the obesity field in search for new treatments for obesity or obesity-related diseases and disorders.

[0003] Diabetes is a group of metabolic disorders characterized by a high blood sugar level. As of 2019, the International Diabetes Federation estimated that 463 million people are suffering from diabetes worldwide, approximately half of the individuals being diagnosed. Diabetes is divided into two types, namely type I and type II diabetes. Type I diabetes results from the pancreas's failure to produce enough insulin due to loss of beta cells caused by an autoimmune response. On the other hand, type II diabetes is a condition that begins with insulin resistance in which cells fail to respond to insulin properly and as the disease progresses may also result in a lack of insulin.

[0004] Human amylin (hAMY 1-37 or amylin) is a 37-residues peptide hormone that is co-secreted with insulin from the pancreatic β-cells with the amino acid sequence Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser-Asn-Asn-Phe-Gly-Ala-Ile-Leu-Ser-Ser-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr with a disulphide bridge between residues 2 and 7. Amylin supresses glucagon release and inhibits gastric emptying and hence plays an important role in maintaining glucose homeostasis by decreasing the blood sugar concentration. Therefore, amylin is a potential candidate for treating diabetes. Furthermore, amylin has been shown to reduce food intake and plays an important role in satiety, also making it a potential candidate for treating obesity. However, amylin possesses some drawbacks such as a high tendency of fibrillation, a short in vivo half-life, and chemical instability at pH 7. Thus, native amylin is suboptimal for use as a pharmaceutically active ingredient.

[0005] A large number of amylin derivatives are known in the prior art, such as the ones disclosed in WO2016 / 146739. These amylin analogues attempt to solve some of the known drawbacks that human amylin possesses. One successful example is the amylin analogue Pramlintide, which has been approved by the FDA for use in type I and type II diabetes. However, Pramlintide is formulated at pH 4 as it fibrillates at pH 7. Thus, there is still a need for new amylin analogues with increased chemical stability, increased metabolic stability, and / or a reduced tendency for fibrillation. In particular, amylin analogues that are stable over a broader pH range are desirable. Furthermore, there is a need for new amylin analogues with increased effectiveness through e.g. increased potency, higher efficacy, and / or longer half-lives to allow less frequent dosing and increased patient compliance. The present invention sets out to solve one or more of the problems known from the prior art by applying a new chemical strategy.SUMMARY OF THE INVENTION

[0006] In a first aspect, the present disclosure relates to hAM 15-52 analogues with improved amylin receptor (hAMY3R) potency (hAMY3R-EC 50 ≤ 250 pM). Thus, in a first aspect the disclosure relates to hAM 15-52 analogues or a pharmaceutically acceptable salt thereof comprising 38 amino acids (X 1 -X 38 ), wherein the amino acid in position X 11 is selected from R, W or Cit and wherein the hAM 15-52 analogue has at least 50 % identity to hAM 15-52 (SEQ ID NO: 1), such as at least 60 % identity to hAM 15-52 (SEQ ID NO: 1). The inventors surprisingly found that the amino acid position X 11 was highly important for increasing amylin receptor potency of hAM 15-52 . Thus, a single substitution in X 11 from K to R, W or Cit was enough to improve the amylin receptor potency (hAMY3R-EC 50 ≤ 250 pM) of hAM 15-52 significantly while retaining adrenomedullin potency (hAM1R).

[0007] In a second aspect, the present invention provides selective hAM 15-52 analogues (hAMY3R-EC 50 ≤ 250 pM and an hAM1R-EC 50 ≥ 25 nM), which act as agonists on the amylin receptor hAMY3R but which, unlike in the prior art (e.g. WO2016 / 146739), are derived from the backbone of the adrenomedullin fragment hAM 15-52 . The inventors surprisingly found that each of the positions X 4 , X 37 and X 38 were highly important for the hAM1R potency of hAM 15-52 and that hAM1R potency of hAM 15-52 could be abolished or reduced (hAMIR-EC 50 ≥ 25 nM) by certain amino acid substitutions in either of the positions X 4 , X 37 and / or X 38 . Thus, in a second aspect, the invention relates to hAM 15-52 analogues comprising 38 amino acids (X 1 -X 38 ) or a pharmaceutically acceptable salt thereof, wherein the amino acid in position X 11 is selected from R, W or Cit and wherein the hAM 15-52 analogue has at least 50 % identity to hAM 15-52 (SEQ ID NO: 1), such as at least 60 % identity to hAM 15-52 (SEQ ID NO: 1) and further wherein X 4 is selected as F, Y, W, T, M, I, A or C; X 37 is selected as G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C or K; X 38 is selected as Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K, with the proviso that at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position. The inventors surprisingly found that the human adrenomedullin fragment hAM 15-52 may be converted into highly selective amylin receptor agonists (hAMY3R-EC 50 value ≤ 250 pM and an hAM1R-EC 50 ≥ 25 nM) by replacing the amino acid in position X 11 in hAM 15-52 together with one or more of the amino acids in the positions X 4 , X 37 and / or X 38 in hAM 15-52 . Thus, replacing at least two amino acids on hAM 15-52 resulted in hAM 15-52 analogues with hAMY3R and hAM1R potencies comparable to amylin rather than hAM 15-52 . One benefit of this approach is that unlike human amylin, hAM 15-52 is not prone to fibrillation, and by using this new chemical strategy, the inventors further envisaged that the good fibrillation properties of hAM 15-52 could be maintained in the new hAM 15-52 analogues. Therefore, the hAM 15-52 analogues according to the invention comprise a significant part of the backbone present in hAM 15-52 as an important part of the sequence (i.e. at least least 50 % identity to hAM 15-52 (SEQ ID NO: 1)). Thus, in the second aspect of the invention, the first problem solved is the provision of new potent hAMY3R agonists with high selectivity over hAM1R, which are obtained by tweaking the polypharmacology of the hAM 15-52 fragment into hAMY3R agonism. The second problem solved by the second aspect is the provision of new hAMY3R agonists which are less prone to fibrillation, thereby overcoming a problem inherent to human amylin. Unlike the amylin derivatives known from the prior art, the hAM 15-52 analogues of the present invention possess a structurally distinct sequence belonging to the hAM 15-52 fragment. Thus, the hAM 15-52 analogues of the invention possess different physical-chemical properties compared to the amylin derivatives in the prior art, such as different solubility, chemical-, physical-, and / or metabolic stability.

[0008] In a third aspect, the invention relates to hAM 15-52 analogues according to the second aspect for use as a medicament. More particularly, the third aspect of the invention relates to hAM 15-52 analogues according to the second aspect for use in treating, preventing or ameliorating a variety of diseases, disorders or conditions, such as but not limited to excess food intake, excess body weight, obesity, Binge eating disorder, Prader-Willi syndrome, dyslipidemia, metabolic diseases / disorders, diabetes I or II, impaired glucose tolerance, insulin resistance syndrome and / or NASH.

[0009] In a fourth aspect, the invention relates to pharmaceutical compositions comprising one or more of the hAM 15-52 analogues according to the second aspect and their medical use(s) in treating, preventing, or ameliorating a variety of diseases, disorders or conditions according to the third aspect. The pharmaceutical compositions may comprise a pharmaceutically acceptable carrier (vehicle) and / or one or more excipient(s).

[0010] The invention will now be explained in more detail from the alignment of the amylin (hAMY 1-37 ) and the adrenomedullin fragment (hAM 15-52 ) as shown in Table 1 below.DEFINITIONS

[0011] In the present context, when the disclosure refers to positions of amino acids in the hAM 15-52 analogues (i.e. X 1 -X 38 ) that are derived / selected from hAMY 1-37 or retained / selected from the hAM 15-52 fragment, the following alignment shown in Table 1 applies. Thus, position X 1 in the hAM 15-52 analogues according to the invention corresponds to the first amino acid in human amylin (hAMY 1-37 ) and amino acid number 15 (hAM 15 ) in the hAM 15-52 fragment.

[0012] According to the present invention, the hAM 15-52 analogues are generally amidated at the C-terminal (-CONH 2 ), like the native peptides; amylin and adrenomedullin. However, the hAM 15-52 analogues of the present invention may also have either a free carboxylic acid (-COOH) or another post-translational modification such as a methyl ester (-COOMe). In a highly preferred embodiment of the invention, the hAM 15-52 analogues are amidated at the C-terminal. The hAM 15-52 analogues according to the present invention may have a free amine (-NH 2 ), be N-acylated (-NHCOR), N-methylated (-NHCH 3 or -N(CH 3 ) 2 ) or deaminated at the N-terminal. The hAM 15-52 analogues may also be lipidated, e.g. at the N-terminal as exemplified herein, depending on the desired half-life of the polypeptides.

[0013] According to the present invention, lipidation has the usual meaning in the art. Thus, lipidation in the present context refers to the covalent attachment of a lipid optionally through a linker to hAM 15-52 analogues of the invention. Lipidation is typically performed to improve the pharmacokinetic profile of a polypeptide by e.g. improving metabolic stability, reducing enzymatic degradation, lowering excretion and metabolism, all in all resulting in a prolonged in vivo half-life (t 1 / 2 ) of the hAM 15-52 analogues. The hAM 15-52 analogues according to the invention may be lipidated or non-lipidated. The lipidated hAM 15-52 analogues exemplified herein are lipidated with a saturated C 20 -diacid through various linkers as shown in table 2. According to the present invention, the abbreviations Hyp, Cit, Aib, Aad, (NMe)G / Sar, (NMe)I have the usual meaning in the art. Thus, Hyp refers to L-hydroxyproline, Cit refers to L-citrulline, Aib refers to 2-aminoisobutyric acid, Aad refers to L-homoglutamic acid, (NMe)G refers to N-methylglycine also known as Sar or sarcosine and (NMe)I refer to N-methyl-L-isoleucine.

[0014] According to the present invention, EC 50 values are used as a measure of agonist potency at a given receptor. An EC 50 value is a measure of the concentration of a compound required to achieve half of that compound's maximal activity in a particular assay. Thus, an hAM 15-52 analogue according to the invention having an hAMY3R-EC 50 lower than the hAMY3R-EC 50 of hAM 15-52 in the given assay is considered to have higher potency or activity at the hAMY3R receptor than the adrenomedullin fragment hAM 15-52 . Likewise, an hAM 15-52 analogue according to the invention having an hAM1R-EC 50 higher than the hAM1R-EC 50 of hAM 15-52 , in the given assay may be considered to have lower potency or activity at the hAM1R receptor than the adrenomedullin fragment hAM 15-52 .

[0015] According to the present invention, an hAM 15-52 analogue or derivative thereof may be in the form of a pharmaceutically acceptable salt. Thus, pharmaceutically acceptable salts are intended to include any salts that are commonly used in formulations of peptides. Such salts include both acid addition salts and basic salts, and examples may be found e.g. in Remington's Pharmaceutical Sciences, 17 th< edition. Likewise, various solvates of the hAM 15-52 analogues or pharmaceutically acceptable salts thereof are also within the scope of the invention.

[0016] According to the present invention, hAM 15-52 analogues are short chains of amino acids that have been linked by amide bonds. In the most preferred embodiment of the invention, the hAM 15-52 analogues are 38 amino acids in length, not including any amino acids that function as spacers between an optional lipid and the hybrid polypeptides.

[0017] In the present context, unless otherwise stated, the amino acids are all L-amino acids (L-stereoisomer, natural amino acids). Thus, the absolute configuration of the amino acids is the (S)-configuration with the exception of L-cysteine and L-selenocysteine having the (R)-configuration.

[0018] In the present context, it should be understood that the amino acid Cys (X 2 ) and Cys (X 7 ) in the hAM 15-52 analogues are covalently connected by a bridge, preferably a disulfide bridge (-S-S-) such as present in hAMY 1-37 and hAM 15-52 . The cyclization of the disulfide bridge may be performed after purification of the hybrid polypeptides or prior to cleavage from the resin in the solid-phase peptide synthesis, optionally in the presence of suitable protecting groups. The disulfide bridge may be formed spontaneously by stirring the hybrid polypeptide in the presence of oxygen or may be formed by treating the hybrid polypeptides with another suitable oxidant such as iodine (I 2 ), optionally in the presence of a base.

[0019] In the present context, the hAM 15-52 analogues or derivatives thereof according to the first or second aspect have an hAMY3R potency (EC 50 ) ≤ 250 pM, such as ≤ 200 pM, preferably an hAMY3R-EC 50 ≤ 150 pM, such as ≤ 125 pM, more preferably an hAMY3R-EC 50 ≤ 100 pM, such as ≤ 75 pM, yet more preferably an hAMY3R-EC 50 ≤ 50 pM, such as ≤ 25 pM, most preferably an hAMY3R-EC 50 ≤ 15 pM, such as ≤ 10 pM. Thus, the most hAMY3R potent hAM 15-52 analogues may be more or less equipotent with native human amylin hAMY 1-37 . For example, SEQ ID NOs: 3-44 have an hAMY3R-EC 50 between 6.0-10 pM and are thus more or equipotent compared to hAMY 1-37 (hAMY3R-EC 50 = 10 pM) when measured under the same assay conditions.

[0020] In the present context, the hAM 15-52 analogues or derivatives thereof according to the second aspect have an abolished or reduced EC 50 when the EC 50 value on hAM1R ≥ 25 nM, such as EC 50 value on hAM1R ≥ 50 nM, such as an hAM1R-EC 50 ≥ 100 nM, preferably an hAM1R-EC 50 ≥ 150 nM, such as ≥ 200 nM, more preferably hAM1R-EC 50 ≥ 250 nM, such as ≥ 300 nM, even more preferably an hAM1R-EC 50 ≥ 350 nM, such as ≥ 400 nM, yet more preferably an hAM1R-EC 50 ≥ 450 nM, such as ≥ 500 nM, yet more preferably an hAM1R-EC 50 ≥ 600 nM, such as ≥ 700 nM, yet more preferably an hAM1R-EC 50 ≥ 800 nM, such as ≥ 900 nM, yet more preferably an hAM1R-EC 50 ≥ 1000 nM, most preferably an hAM1R-EC 50 ≥ 5000 nM. Thus, in the present context, selectively in terms of potency towards the amylin receptor hAMY3R over the adrenomedullin receptor hAM1R (i.e. a selective amylin receptor agonist) should be understood as an hAM 15-52 analogue or a derivative thereof according to the second aspect having an hAMY3R-EC 50 value ≤ 250 pM and an hAM1R-EC 50 ≥ 25 nM. Thus, a selective hAM 15-52 analogue or a derivative thereof has a selectivity ratio (hAM1R EC 50 / hAMY3R EC 50 ) of at least 100. Preferably, the hAM 15-52 analogues or derivatives thereof have an hAMY3R-EC 50 value ≤ 250 pM and an hAM1R-EC 50 ≥ 50 nM and thus a selectivity ratio (hAM1R EC 50 / hAMY3R EC 50 ) ≥ 200. More preferably, the hAM 15-52 analogues or derivatives thereof have an hAMY3R-EC 50 ≤ 200 pM, an hAM1R-EC 50 ≥ 100 nM and thus a selectivity ratio (hAM1R EC 50 / hAMY3R EC 50 ) ≥ 500. More preferably, an hAM 15-52 analogue or a derivative thereof has an hAMY3R-EC 50 ≤ 125 pM, an hAM1R-EC 50 ≥ 200 nM and thus a selectivity ratio (hAM1R EC 50 / hAMY3R EC 50 ) ≥ 1600. Even more preferably, the hAM 15-52 analogue or a derivative thereof has an hAMY3R-EC 50 ≤ 100 pM, an hAM1R-EC 50 ≥ 500 nM and thus a selectivity ratio (hAM1R EC 50 / hAMY3R EC 50 ) ≥ 5000. Most preferably, the hAM 15-52 analogue or a derivative thereof has an hAMY3R-EC 50 ≤ 50 pM, an hAM1R-EC 50 ≥ 1000 nM and thus a selectivity ratio (hAM1R EC 50 / hAMY3R EC 50 ) ≥ 20000. For example, any of SEQ ID NOs: 3-42 have a selectivity ratio of at least 6000 and most of them much higher.

[0021] A person skilled in the art is well aware that EC 50 values depend on the assay type and assay conditions. Thus, in the present context when the disclosure refers to an EC 50 value, it should be understood as an EC 50 value obtained when measured according to the procedure described in the examples under general protocols for cAMP assays for measuring in vitro receptor activation. However, variation may also be present within the same assay under apparently identical assay conditions due to variation in e.g. receptor expression in the cells (i.e. receptor density). Thus, in order to compare EC 50 values, hAMY 1-37 (SEQ ID NO: 2) and hAM 15-52 (SEQ ID NO: 1) have been tested as internal standards for comparison between different assay runs or even different assays. The hAMY3R-EC 50 and the hAM1R-EC 50 of hAMY 1-37 (SEQ ID NO: 2) was determined to 10 pM and 5000 nM, respectively, using the same assay conditions and cell line as the hAM 15-52 analogues (SEQ ID NO: 3-392). Likewise, the hAMY3R-EC 50 and the hAM1R-EC 50 of the hAM 15-52 (SEQ ID NO: 1) was determined to 1.3 nM and 1.1 nM, respectively, using the same assay conditions and cell line as the hAM 15-52 analogues (SEQ ID NO: 3-392). Thus, it follows that the hAM 15-52 analogues or derivatives thereof according to the first or second aspect have a relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAMY 1-37 -hAMY3R-EC 50 ) ≤ (250 pM) / (10 pM) ≤ 25. Preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAMY 1-37 -hAMY3R-EC 50 ) ≤ 20. More preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAM 1-37 -hAMY3R-EC 50 ) ≤ 12.5. Even more preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAMY 1-37 -hAMY3R-EC 50 ) ≤ 10. Even more preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAMY 1-37 -hAMY3R-EC 50 ) ≤ 5. Yet more preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAMY 1-37 -hAMY3R-EC 50 ) ≤ 2. Most preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAMY 1-37 -AMY3R-EC 50 ) ≤ 1 such that the hAM 15-52 analogues or derivatives thereof an equipotent or more potent than hAMY 1-37 . Likewise, the selective hAM 15-52 analogues or derivatives thereof according to the second aspect have a relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAM 15-52 -AMY3R-EC 50 ) ≤ (250 pM) / (1300 pM) ≤ 0.19 and a relative hAM1R-EC 50 ratio (hAMIR-EC 50 ) / (hAM 15-52 -hAMIR-EC 50 ) ≥ (25 nM) / (1.1 nM) ≥ 22.7. Preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAM 15-52 -hAMY3R-EC 50 ) ≤ (250 pM) / (1300 pM) ≤ 0.19 and a relative hAM1R-EC 50 ratio (hAMIR-EC 50 ) / (hAM 15-52 -hAMIR-EC 50 ) ≥ (50 nM) / (1.1 nM) ≥ 45.5. More preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAM 15-52 -hAMY3R-EC 50 ) ≤ 0.15 and a relative hAM1R-EC 50 ratio (hAM1R-EC 50 ) / (hAM 15-52 -hAM1R-EC 50 ) ≥ 76.9. Yet more preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAM 15-52 -hAMY3R-EC 50 ) ≤ 0.096 and a relative hAM1R-EC 50 ratio (hAMIR-EC 50 ) / (hAM 15-52 -hAMIR-EC 50 ) ≥ 154. Even more preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAM 15-52 -hAMY3R-EC 50 ) ≤ 0.077 and a relative hAM1R-EC 50 ratio (hAMIR-EC 50 ) / (hAM 15-52 -hAMIR-EC 50 ) ≥ 384. Yet more preferably, the relative hAMY3R-EC 50 ratio (hAMY3R-EC 50 ) / (hAM 15-52 -AMY3R-EC 50 ) ≤ 0.038 and a relative hAM1R-EC 50 ratio (hAM1R-EC 50 ) / (hAM 15-52 -hAM1R-EC 50 ) ≥ 769. Most preferably, the relative hAM1R-EC 50 ratio (hAM1R-EC 50 ) / (hAM 15-52 -hAM1R-EC 50 ) ≥ 2000.

[0022] According to the present invention, the term "treatment" should be understood in the broadest sense as prevention, amelioration, or treatment. Thus, treatment is also intended to include prophylactic treatment.

[0023] According to the present invention, from the aspect(s) / embodiment(s) which refer to "wherein hAM 15-52 analogue has (e.g.) at least 50 % identity to hAM 15-52 (SEQ ID NO: 1)" or "a derivative thereof with (e.g.) at least 50 % identity to hAM 15-52 ", it should be understood that the hAM 15-52 analogue or derivative thereof has at least 50 % sequence identity with hAM 15-52 (SEQ ID NO: 1) when aligned. As an example, SEQ ID NO: 169 illustrated below differs in two amino acids (X 11 and X 38 ) from hAM 15-52 (SEQ ID NO: 1) and thus has (38-2) / 38 = 94.7 % sequence identity to hAM 15-52 (SEQ ID NO: 1). SEQ ID NO:1G C R F G T C T V Q K L A H Q I Y Q F T D K D K D N V A P R S K I S P Q G YDerivative X:G C R F G T C T V Q R L A H Q I Y Q F T D K D K D N V A P R S K I S P Q G Hyp DETAILED DESCRIPTION OF THE INVENTION The calcitonin peptide family

[0024] The calcitonin family of peptides consists of the hormone peptides calcitonin (CT), calcitonin gene-related peptide (CGRP), islet amyloid polypeptide (IAPP, amylin or hAMY 1-37 ), and adrenomedullin (hAM) as well as their precursors. hAMY 1-37 is a 37-residues peptide hormone that is co-secreted with insulin from the pancreatic β-cells with the amino acid sequence Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser-Asn-Asn-Phe-Gly-Ala-Ile-Leu-Ser-Ser-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr. Amylin supresses glucagon release and inhibits gastric emptying and hence plays an important role in maintaining glucose homeostasis by decreasing the blood sugar concentration. Furthermore, amylin has shown to reduce food intake and plays an important role in satiety, making it a potential candidate for treating e.g. obesity and / or diabetes. hAM is a 52-residues peptide hormone expressed in all tissues with the amino acid sequence Tyr-Arg-Gln-Ser-Met-Asn-Asn-Phe-Gln-Gly-Leu-Arg-Ser-Phe-Gly-Cys-Arg-Phe-Gly-Thr-Cys-Thr-Val-Gln-Lys-Leu-Ala-His-Gln-Ile-Tyr-Gln-Phe-Thr-Asp-Lys-Asp-Lys-Asp-Asn-Val-Ala-Pro-Arg-Ser-Lys-Ile-Ser-Pro-Gln-Gly-Tyr. It is a potent vasodilator and has shown positive influence in cardiovascular diseases, such as myocardial infarction, limb ischemia and hypertension.

[0025] The biological activity of the calcitonin protein family is generally mediated via binding to the calcitonin receptor (CTR) and the calcitonin receptor like receptor (CRLR), both belonging to family 2 of the G-protein-coupled receptors (GPCR). These receptors may be co-expressed in combination with different receptor modifying proteins (RAMP1-3) generating functional receptors for the individual peptides in the calcitonin protein family. Co-expression of CTR with RAMP1 leads to formation of a receptor for amylin and CGRP (AMY1R), co-expression of CTR with RAMP2 leads to the amylin receptor 2 (AMY2R) and co-expression of CTR with RAMP3 leads to the amylin receptor (AMY3R). Co-expression of CRLR with RAMP1 leads to a formation of a receptor for CGRP (CGRP1R), co-expression of CRLR with RAMP2 leads to a formation of a receptor for adrenomedullin (AM1R) and co-expression of CRLR with RAMP3 leads to a formation of a receptor for adrenomedullin and CGRP (AM2R).

[0026] Several of the native peptides in the calcitonin protein family show considerable overlap in pharmacology between receptors. For example, adrenomedullin is approximately 100 times less potent on AMY3R compared to hAMY 1-37 . The adrenomedullin fragment (hAM 15-52 ) is almost equipotent on AMY3R and AM1R with an EC 50 of 1.3 nM on AMY3R and an EC 50 value of 1.1 nM on AM1R (said EC 50 value being measured according to the examples herein). hAMY 1-37 on the other hand has an EC 50 value of 10 pM on AMY3R while being inactive on AM1R.Determination of important amino acid positions in hAM 15-52

[0027] The inventors initially hypothesized that since hAM 15-52 and hAMY 1-37 belong to the same family of hormones, the Calcitonin / CGRP / IAPP family, the AMY3R potency of hAM 15-52 could be improved or even converted into selective AMY3R agonism over hAM1R agonism, by substituting certain amino acids in hAM 15-52 with amino acids derived from hAMY 1-37 , thereby overcoming some of the drawbacks that are inherent to hAMY 1-37 . All 37 hAMY 1-37 amino acids were systematically substituted into the hAM 15-52 . Furthermore, X 38 was selected as Hyp (Hydroxyproline), which is neither present in hAMY 1-37 nor in hAM 15-52 . The inventors constructed random forest models (Breiman, L. (2001), Random Forests, Machine Learning 45(1), 5-32.) describing the relationship between peptides sequence and hAMY3R and the hAM1R potency, respectively, and systematically synthesised and screened peptide sequences. For each hAM 15-52 analogue, the amino acid in each sequence position were used as features, and receptor EC 50 values were used as response to construct the models. The "randomForest" package implemented in the statistical programming language R was used to calculate the importance of each position and the most important positions were identified to be critical for hAMY3R and / or hAM1R activity (A. Liaw and M. Wiener (2002). Classification and Regression by randomForest. R News 2(3), 18-22.). Based on this systematic analysis the inventors identified X 4 , X 37 and X 38 to be important for hAM1R activity, while of less importance for hAMY3R activity. The inventors identified X 11 to be critical for hAMY3R activity, while of less importance for hAM1R activity. Finally, the inventors identified X 17 to be of some importance for hAMY3R activity, while of less importance for hAM1R activity. Thus, the inventors identified X 11 as a key position for improving hAMY3R potency in hAM 15-52 and X 4 , X 37 and X 38 as key positions for eliminating hAM1R potency in hAM 15-52 . Since X 17 was only of some importance for hAMY3R activity, this position was not considered a key position but only a preferred position to further improve the AMY3R potency of the hAM 15-52 analogues.

[0028] The systematic investigation showed that the hAMY3R potency of hAM 15-52 could be improved by substituting the amino acid K present in hAM 15-52 in position X 11 with the amino acid R present in hAMY 1-37 . Furthermore, the systematic investigation showed that the hAM1R potency of hAM 15-52 could be decreased by substituting the amino acids present in position X 4 , X 37 and X 38 in hAM 15-52 with the corresponding amino acids found in present in hAMY 1-37 .Deeper investigation of amino acids in the position X 4 , X 11 , X 17 , X 37 and X 38 .

[0029] The inventors systematically substituted representative amino acids covering representative amino acids for all the known amino acids into the 5 identified positions, at one position at the time (Table 6). As shown in example 1, all (SEQ ID: 1-360 had lost their activity on the hAM1R, with varying degree of potency on AMY3R. In order to score the individual amino acids on the 5 positions, the average EC 50 value for peptides containing a given amino acid in a given position was compared to the average EC 50 value hAM 15-52 analogues containing the reference amino acid in that position. Given a cut-off of 4-fold changes in potency, each amino acid in each position was scored as giving increased (I), decreased (D) or unchanged (U) potency, relative to the reference amino acid, on hAMY3R. The results are summarized in Table 6. From this data, it was shown that any of the amino acids R, W or Cit were capable of maintaining the AMY3R potency when present in position X 11 when compared to the reference whereas other representative amino acids decreased the potency. It was further shown that any of the amino acids Y, W, T, M, I, F, A, or C in X 4 , any of the amino acids Y, W, T, Q, P, M, I, H, F, E, A, R, C or K in X 37 and any of the amino acids Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K in X 38 were able to maintain or improve AMY3R potency while remaining inactive on hAM1R.Aspect 1: hAM 15-52 analogues with improved amylin potency

[0030] Thus, in a first aspect, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising 38 amino acids (X 1 -X 38 ) with an hAMY3R-EC 50 ≤ 250 pM, wherein the amino acid in position X 11 is selected from R, W or Cit and wherein the hAM 15-52 analogue has at least 50% identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , (SEQ ID NO: 1). From example 1 (Table 2), it can be seen that almost all of the synthesized hAM 15-52 analogues have at least 50 % identity to hAM 15-52 , that a vast majority of the synthesized hAM 15-52 analogues have at least 60 % of identity to hAM 15-52 , and that a large portion of the synthesized hAM 15-52 analogues have at least 80 % identity to hAM 15-52 (SEQ ID NO: 1). All of these hAM 15-52 analogues maintain the good fibrillation properties of hAM 15-52 (SEQ ID NO: 1) compared to hAMY 1-37 (SEQ ID NO: 2) (see example 2, Table 3). E.g., SEQ ID NO: 32 shows that the good physical stability (i.e. low fibrillation) of hAM 15-52 can be maintained as long as the hAM 15-52 analogues have at least 50 % identity to hAM 15-52 .

[0031] Thus, in a first aspect, the disclosure relates to a way to highly improve the hAMY3R potency of the hAM 15-52 fragment by a single substitution to R, W or Cit in position X 11 , while maintaining the good physical stability (i.e. good fibrillation properties of hAM 15-52 ). These hAM 15-52 analogues will therefore be highly potent agonists on both hAMY3R and hAM1R (see SEQ ID NO: 361-392).

[0032] In the most preferred embodiment of the disclosure, X 11 is selected as R. In another embodiment, X 11 is selected as W. In yet an embodiment, X 11 is selected as Cit. The inventors found that this single amino acid substitution in X 11 provided increased hAMY3R potency to the hAM 15-52 analogue(s) while retaining hAM1R potency. The inventors further found that the hAMY3R potency could be maintained when X 17 was Y, T, Q, M, I, F, A, R or K, or even further improved by selecting X 17 as W or H. Thus, in an embodiment X 17 is selected as Y, W, T, Q, M, I, H, F, A, R or K. In a preferred embodiment, X 17 is selected as Y, W or H, more preferably W or H.

[0033] In an embodiment, the disclosure relates to an hAM 15-52 analogue comprising 38 amino acids (X 1 -X 38 ) or a pharmaceutically acceptable salt thereof, wherein the amino acid in position X 11 is selected from R, W or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R or K, preferably X 17 is selected as W or H and wherein the hAM 15-52 analogue has at least 50 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 (SEQ ID NO: 1).

[0034] In yet an embodiment, the disclosure relates to an hAM 15-52 analogue comprising 38 amino acids (X 1 -X 38 ) or a pharmaceutically acceptable salt thereof, wherein the amino acid in position X 11 is R; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R or K, preferably X 17 is selected as W or H and wherein the hAM 15-52 analogue has at least 50% identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , (SEQ ID NO: 1).

[0035] In yet an embodiment, the invention relates to an hAM 15-52 analogue comprising 38 amino acids (X 1 -X 38 ) or a pharmaceutically acceptable salt thereof, wherein the amino acid in position X 11 is W; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R or K, preferably X 17 is selected as W or H and wherein the hAM 15-52 analogue has at least 50% identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , (SEQ ID NO: 1).

[0036] In yet an embodiment, the disclosure relates to an hAM 15-52 analogue comprising 38 amino acids (X 1 -X 38 ) or a pharmaceutically acceptable salt thereof, wherein the amino acid in position X 11 is Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R or K, preferably X 17 is selected as W or H and wherein the hAM 15-52 analogue has at least 50% identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , (SEQ ID NO: 1).

[0037] In a preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K.

[0038] In another preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K.

[0039] In a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K.

[0040] In yet a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K.

[0041] In yet a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , X 11 is selected from R; X 17 is selected as Y, W or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K.

[0042] In yet a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F.

[0043] In yet a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K.

[0044] In yet a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F.

[0045] In yet a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, M, or F; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, M, or F.

[0046] In yet an even more preferred embodiment, the disclosure relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R; X 17 is selected as Y, W, or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, M, or F; or a derivative thereof with at least 50% identity to hAM 15-52 , preferably at least 60% identity to hAM 15-52 , such as at least 70% identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , even more preferably at least 85 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R; X 17 is selected as Y, W or H; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, M, or F.

[0047] In yet a more preferred embodiment, the disclosure relates to an hAM 15-52 analogue comprising the amino acid sequence of formula (I): wherein, X 11 is selected from R, W, or Cit or a derivative thereof with at least 50% identity to hAM 15-52 , least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52 , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , most preferably at least 90 % identity to hAM 15-52 to hAM 15-52 , such as at least 95 % identity to hAM 15-52 , wherein, X 11 is selected from R, W, or Cit.

[0048] In a most preferred embodiment, the disclosure relates to an hAM 15-52 analogue of SEQ ID NO: 3-392 or a derivative thereof with at least 90 % identity to any one of SEQ ID NO: 3-392, preferably at least 95 % identity any one of SEQ ID NO: 3-392, more preferably at least 96 % identity any one of SEQ ID NO: 3-392 most preferably at least 97 % identity any one of SEQ ID NO: 3-392 with the proviso that a derivative thereof has R, W, or Cit in position X 11 , preferably R in position X 11 .

[0049] In yet a most preferred embodiment, the disclosure relates to an hAM 15-52 analogue of SEQ ID NO: 3-392 or a derivative thereof with at least 90 % identity to any one of SEQ ID NO: 3-392, preferably at least 95 % identity to any one of SEQ ID NO: 3-392, more preferably at least 96 % identity any one of SEQ ID NO: 3-392, most preferably at least 97 % identity any one of SEQ ID NO: 3-278, with the proviso that a derivative thereof has the same amino acid in position X 11 , as the respective SEQ ID NO: 3-392 from it is derived.Aspect 2: Selective hAM 15-52 analogues with improved amylin potency Number of positions needed to abolish or reduce AM1R potency in hAM 15-52 .

[0050] The inventors found that hAM1R potency could be abolished or reduced in hAM 15-52 by changing at least one amino acid in any of the positions X 4 , X 37 or X 38 since all of them were critical for hAM1R potency.Decreasing hAM1R potency with amino acids in position X 4 , X 37 or X 38 .

[0051] X 4 : The inventors found that the amino acids Y, W, T, M, I, A or C in position X 4 were capable of decreasing the hAM1R potency. Thus, in an embodiment, X 4 is selected from Y, W, T, M, I, A or C. Furthermore, the amino acids W, M, I or C in position X 4 were most effective at improving hAMY3R potency (see example 5, table 6). Thus, in a highly preferred embodiment of the invention, X 4 is selected as W, M, I or C. Since the inventors found that a single position chosen among X 4 , X 37 or X 38 was enough to abolish or reduce AM1R potency it follows that in some embodiments X 4 is F (i.e. the native amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position) and the AM1R potency is abolished or reduced using position X 37 and / or X 38 . Thus, in any of the above embodiments for X 4 , F may be included and the AM1R potency abolished or reduced using position X 7 and / or X 38 .

[0052] X 37 : Likewise, the inventors found that the amino acids Y, S, W, T, Q, P, M, I, H, F, E, A, R, C in position X 37 were capable of decreasing the hAM1R potency. Thus, in an embodiment, X 37 is selected from Y, S, W, T, Q, P, M, I, H, F, E, A, R, C. Furthermore, the amino acids Y, W, P, H or F in position X 37 were most effective at improving hAMY3R potency (see example 5, table 6). Thus, in a highly preferred embodiment of the invention, X 37 is selected as Y, W, P, H or F. Since the inventors found that a single position chosen among X 4 , X 37 or X 38 was enough to abolish or reduce AM1R potency it follows that in some embodiments X 37 is G (i.e. the native amino present in hAM 15-52 (SEQ ID NO: 1) in said position), and the AM1R potency is abolished or reduced using position X 4 and / or X 38 . Thus, in any of the above embodiments for X 37 , G may be included and the AM1R potency abolished or reduced using position X 4 and / or X 38 .

[0053] X 38 : Likewise, the inventors found that the amino acids Hyp, W, T, Q, P, M, I, H, F, E, A, R, or K in position X 38 were capable of decreasing the hAM1R potency. Thus, in an embodiment, X 38 is selected from Hyp, W, T, Q, P, M, I, H, F, E, A, R, or K. Furthermore, the amino acids Hyp, W, M or F were most effective at improving hAMY3R potency (see example 5, table 6). Thus, in a highly preferred embodiment of the invention, X 38 is selected as Hyp, W, M or F. In the most preferred embodiment of the invention, X 38 is selected as Hyp. Since the inventors found that a single position chosen among X 4 , X 37 or X 36 was enough to abolish or reduce AM1R potency it follows that in some embodiments X 38 is Y (i.e. the native amino present in hAM 15-52 (SEQ ID NO: 1) in said position), and the AM1R potency is abolished or reduced using position X 4 and / or X 37 . Thus, in any of the above embodiments for X 38 , Y may be included and the AM1R potency abolished or reduced using position X 4 and / or X 37 .

[0054] A simple illustration of aspect 2 of the invention can be seen by comparing the alignment of hAM 15-52 (SEQ ID NO: 1) and SEQ ID NO: 169 shown below. SEQ ID NO:1G C R F G T C T V Q K L A H Q I Y Q F T D K D K D N V A P R S K I S P Q G YSEQ ID NO:169G C R F G T C T V Q R L A H Q I Y Q F T D K D K D N V A P R S K I S P Q G HypSEQ ID NO:1hAMY3R EC 50 1.274 nM and hAM1R EC 50 1.113 nMSEQ ID NO:169hAMY3R EC 50 0.063 nM and hAM1R EC 50 2981.0 nM

[0055] As can be seen, the R in position X 11 has improved the potency on AMY3R approximately 20-fold, and the Hyp in position X 38 has decreased the potency on AM1R approximately 2500-fold. Therefore, only two substitutions have converted hAM 15-52 into an amylin agonist with same pharmacological profile as hAMY 1-37 . At the same time, SEQ ID NO: 169 has maintained the physical stability (ThT signal: 2.700 %) of hAM 15-52 (ThT signal: 2.600 %) compared to hAMY 1-37 (ThT signal: 100.0 %) (see example 2, Table 3). Thus, in a second aspect, the invention relates to a way to more or less completely abolish hAM1R potency of the hAM 15-52 fragment by one or more substitutions in position X 4 , X 37 and / or X 38 and at the same time improve or maintain sufficient AMY3R potency. These hAM 15-52 analogues will therefore be highly potent and selective hAMY3R analogues. At the same time these hAM 15-52 analogues maintain the physical stability of hAM 15-52 due to the sequence identity with hAM 15-52 . As shown in example 2, Table 3, the good physical stability of hAM 15-52 may be maintained or even improved in the peptides (due to at least 50 % identity to hAM 15-52 ), while providing new hAMY3R agonists that are as potent and selective as natural human amylin. In other words, the second aspect of the present invention has created a functional human amylin analogue disguised in an adrenomedullin backbone.Increasing hAMY3R potency with amino acids in position X 17 .

[0056] The inventors further found that the hAMY3R potency could be maintained when X 17 as Y, T, Q, M, I, F, A, R, or K, or even further improved when X 17 was W or H. Thus, in an embodiment, X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K. In a preferred embodiment, X 17 is selected as W, or H.

[0057] Thus, in a second aspect, the invention relates to hAM 15-52 analogues comprising 38 amino acids (X 1 -X 38 ) or a pharmaceutically acceptable salt thereof, wherein the amino acid in position X 11 is selected from R, W or Cit and wherein the hAM 15-52 analogue has at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , most preferably at least 90 % identity to hAM 15-52 , such as at least 95 % identity to hAM 15-52 (SEQ ID NO: 1) and further wherein X 4 is selected as F, Y, W, T, M, I, A or C; X 37 is selected as G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C or K; X 38 is selected as Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K, with the proviso that at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position, more preferably, at least two of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position. In the second aspect, the hAM 15-52 analogues have an hAMY3R-EC 50 ≤ 250 pM and an hAM1R-EC 50 ≥ 25 nM.

[0058] In an embodiment, X 4 is selected as F, W, M, I or C; X 37 is selected as G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected as Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K, with the proviso that at least one of the positions X 4 , X 37 , or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position, more preferably, at least two of the positions X 4 , X 37 , or X 38 are not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0059] In an embodiment, X 4 is selected as F, Y, W, T, M, I, A or C; X 37 is selected as G, Y, W, P, H or F; X 38 is selected as Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K, with the proviso that at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position, more preferably, at least two of the positions X 4 , X 37 or X 38 are not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0060] In an embodiment, X 4 is selected as F, Y, W, T, M, I, A or C; X 37 is selected as G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C or K; X 38 is selected as Hyp, Y, W, M or F, with the proviso that at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position, more preferably, at least two of the positions X 4 , X 37 , or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0061] In an embodiment, X 4 is selected as F, W, M, I, or C; X 37 is selected as G, Y, W, P, H, or F; X 38 is selected as Hyp, Y, W, M or F, with the proviso that at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position, more preferably, at least two of the positions X 4 , X 37 , or X 38 are not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0062] In a preferred embodiment, the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity, even more preferably at least 85 % identity, such as at least 90 % identity, most preferably at least 95 % identity, wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, K; X 4 is selected from F, Y, W, T, M, I, A or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0063] In yet a more preferred embodiment, the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R or K; X 4 is selected from F, W, M, I or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity, such as at least 85 % identity, more preferably at least 90 % identity, most preferably at least 95 % identity, wherein X 11 is selected from R, W or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R or K; X 4 is selected from F, W, M, I or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0064] In another more preferred embodiment, the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity, such as at least 85 % identity, more preferably at least 90 % identity, most preferably at least 95 % identity, wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 , or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0065] In yet a more preferred embodiment, the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity, such as at least 85 % identity, more preferably at least 90 % identity, most preferably at least 95 % identity, wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 , or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0066] In yet a more preferred embodiment, the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , more preferably at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0067] In a preferred embodiment of the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , more preferably at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0068] In a preferred embodiment, the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity, such as at least 85 % identity to hAM 15-52 , more preferably at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A or C; X 37 is selected from G, Y, W, P, H or F; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 , or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0069] In a more preferred embodiment, the invention relates to an hAM 15-52 analogue or a pharmaceutically acceptable salt thereof comprising the amino acid sequence of formula (I): wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 85 % identity to hAM 15-52 , more preferably at least 90 % identity to hAM 15-52 , most preferably at least 95 % identity to hAM 15-52 , wherein X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, W, M, I, or C; X 37 is selected from G, Y, W, P, H, or F; X 38 is selected from Hyp, Y, W, M, or F; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0070] In any of the above-mentioned embodiments, X 11 is most preferably R, and / or X 17 is most preferably Y, W, or H. Furthermore, in any of the above-mentioned embodiments, most preferably at least two of the positions X 4 , X 37 or X 38 are not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0071] In a more highly preferred embodiment, the invention relates to an hAM 15-52 analogue with the sequence SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360 or a derivative thereof with at least 50 % identity to hAM 15-52 , such as at least 55 % identity to hAM 15-52 , such as at least 60 % identity to hAM 15-52 , such as at least 65 % identity to hAM 15-52 , preferably at least 70 % identity to hAM 15-52 , such as at least 75 % identity to hAM 15-52, , more preferably at least 80 % identity to hAM 15-52 , such as at least 90 % identity to hAM 15-52 (SEQ ID NO: 1), wherein, in the derivative, X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A, or C; X 37 is selected from G, Y, S, W, T, Q, P, Mv, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0072] In yet a more highly preferred embodiment, the invention relates to an hAM 15-52 analogue with the sequence SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360 or a derivative thereof with at least 90 % identity to any one of SEQ ID NO: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, preferably at least 95 % identity to any one of SEQ ID NO: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, most preferably at least 97 % identity to anyone of SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, with the proviso that in the derivative, X 11 is selected from R, W, or Cit; X 17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X 4 is selected from F, Y, W, T, M, I, A or C; X 37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X 38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0073] In any of the above-mentioned embodiments, X 4 is preferably selected as F, W, M, I, or C; X 11 is preferably selected as R; X 37 is preferably selected as G, Y, W, P, H, or F; X 38 is preferably selected as Hyp, Y, W, M, or F; X 17 is preferably selected as W or H. Furthermore, in any of the above-mentioned embodiments, most preferably at least two of the positions X 4 , X 37 or X 38 is not the amino acid present in hAM 15-52 (SEQ ID NO: 1) in said position.

[0074] In the most preferred embodiment, the invention relates to an hAM 15-52 analogue with the sequence SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360 or a derivative thereof with at least 90 % identity to anyone of SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, preferably at least 95 % identity to anyone of SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, most preferably at least 97 % identity to anyone of SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, with the proviso that a derivative thereof has the same amino acids in positions X 4 , X 11 , X 37 , and X 38 , preferably the same amino acids in position X 4 , X 11 , X 17 , X 37 , and X 38 , as the respective hAM 15-52 analogue of SEQ ID NO: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360 from which it is derived.Medical use

[0075] In a third aspect, the invention relates to hAM 15-52 analogues according to the second aspect for use as a medicament. More particularly, the third aspect of the invention relates to hAM 15-52 analogues according to the second aspect for use in treating, preventing or ameliorating a variety of diseases, disorders or conditions, such as but not limited to excess food intake, excess body weight, obesity, Binge eating disorder, Prader-Willi syndrome, dyslipidemia, metabolic diseases / disorders, diabetes I or II, impaired glucose tolerance, insulin resistance syndrome and / or NASH. A preferred embodiment relates to one or more hAM 15-52 analogue(s) for use in the treatment of obesity, diabetes, NASH or combinations thereof.Pharmaceutical compositions

[0076] In a fourth aspect, the invention relates to pharmaceutical compositions comprising one or more of the hAM 15-52 analogues according to the second aspect and their medical use(s) in treating, preventing, or ameliorating a variety of diseases, disorders or conditions according to the third aspect. The pharmaceutical compositions may comprise a pharmaceutically acceptable carrier (vehicle) and / or one or more excipient(s) in accordance with conventional techniques in the art, such as those disclosed in 'Remington: Essentials of Pharmaceutics', Ed. by Linda A. Felton, Pharmaceutical press 2012.

[0077] Suitable formulations include but are not limited to tablets, pills, capsules, emulsions, suspensions, sustained release, or solutions. It should be appreciated that different routes of administration may be used depending on the choice of formulation and chemical and / or metabolic stability of the polypeptides. Such administration routes may include but are not limited to oral administration, parenteral administration (intravenous (IV), subcutaneous (SC), intradermal (ID) and intramuscular (IM)), or inhalation. In a preferred embodiment of the invention, the administration route is parental administration. In an even more preferred embodiment, the administration route is subcutaneous.EXAMPLESGeneral protocols for synthesis of the hybrid polypeptides

[0078] The peptides were synthesized using a SyroII fully automated parallel peptide synthesizer (MultiSynTech GmbH, Germany), equipped with heating block, on Tentagel 5 RAM with a loading of 0.23-0.25 mmol / g (Rapp polymer GmbH, Germany). N α< -Fmoc deprotection was performed in two stages by treating the resin with 40 % piperidine / DMF (0.2 M HOBt (1-hydroxybenzotriazole)) for 3 min at 45°C followed by 20 % piperidine / DMF (0.1 M HOBt) for 7-12 min at 75°C. Except Asp, Cys and His residues which were N α< -Fmoc deprotections at room temperature; i.e. 40 % piperidine / DMF (0.2 M HOBt) for 3 min followed by 20 % piperidine / DMF (0.1 M HOBt) for 15 min. The coupling chemistry was DIC (N,N'-diisopropylcarbodiimide) / Oxyma (ethyl cyano(hydroxyimino)acetate) in DMF using amino acid solutions of 0.5 M in DMF and a molar excess of 6-fold. Standard Fmoc protected amino acids were used. Coupling conditions was single or double couplings for 15 min at 75°C. Except His and Cys residues, which were double coupled for 15 min at 50°C. The Fmoc-amino acids were dissolved at 0.5 M in DMF containing 0.5 M Oxyma, except His which was dissolved in NMP. The resin was washed 4x with NMP after N α< -Fmoc deprotection and 3x after couplings.

[0079] The disulfide bridge was formed on the resin by using Trityl (Trt) as the protecting group for cysteine and treating with 1 % iodine in 75 % HFIP (Hexafluoroisopropanol) in DCM for 1 min. The resin was washed 3x with 75 % HFIP in DCM followed by 4x DCM.

[0080] After synthesis, the resin was washed with DCM and dried, and the polypeptide was cleaved from the resin by a 35 min treatment with TFA (trifluoroacetic acid) / TES (triethylsilane) / water (95 / 2.5 / 2.5) at 42°C, followed by precipitation with 4 volumes of cold diethyl ether, further washing with diethyl ether and left to dry.

[0081] The peptides were characterized by LC-MS (Waters, Denmark) and quantified by LC-CAD (ThermoFisher scientific, Denmark). Finally, the peptides were freeze dried to give a white powder using a Telstar benchtop freeze drier.General protocols for cAMP assays for measuring in vitro receptor activation hAMY3-R:

[0082] Cells stably overexpressing the hAMY3 receptor were obtained from Ogeda (now Astellas Pharma), subcloned and a monoclonal cell-line with an appropriate assay-window was expanded, aliquoted and frozen. An aliquot was thawed and plated in DPBS with 0.05 % casein and 0.5 mM IBMX as 2000 cells / well in a 384-well format. The cells were then immediately stimulated for 30 min at room temperature with graded doses of test compound using human amylin (Bachem, cat no. H-7905) as a positive control. cAMP accumulation was measured using a Cisbio assay for Gs coupled receptors (cat. no. 62AM4PEC), where the assay reagents were added as per the manufacturer's instructions and time-resolved fluorescence energy transfer recorded after one hour.hAM1-R:

[0083] Cells stably overexpressing the hAM1 receptor were obtained from Ogeda (now Astellas Pharma), subcloned and a monoclonal cell-line with an appropriate assay-window was expanded, aliquoted and frozen. An aliquot was thawed and plated in DPBS with 0.05 % casein and 0.5 mM IBMX as 8000 cells / well in a 384-well format. The cells were then immediately stimulated for 30 min at room temperature with graded doses of test compound using human adrenomedullin 1-52 (Bachem, cat no. H-2932) as a positive control. cAMP accumulation was measured using a Cisbio assay for Gs coupled receptors (cat. no. 62AM4PEC), where the assay reagents were added as per the manufacturer's instructions and time-resolved fluorescence energy transfer recorded after one hour.General protocols for determination of physical stability of peptide analogues

[0084] Peptides were dissolved in buffers (50 mM sodium acetate at pH 4 or 50 mM sodium phosphate at pH 7) and incubated for one hour. The samples were then divided into two replicates of 80 µl in a black 384 well plate (µ-clear, Greiner Bio-One) and mixed with Thioflavin T (ThT) to a final concentration of 4 µM. The plate was sealed and placed in a plate reader (CLARIOstar, BMG) where the temperature was set at 40°C during the course of the experiment. The samples were further stressed by shaking the plate at 700 rpm (linear) for five minutes before every measurement. The fluorescence was measured every 10 min for four days by exciting the ThT at 450 nm and measuring the emission at 480 nm. For each peptide, the ThT signal over time was smoothened using Local Polynomial Regression fitting (LOESS) as implemented in the statistical programming environment R. For the smoothened data, the maximum ThT signal was normalized in percent between the maximum ThT signal for hAMY and the buffer background. Thus, high values indicate fibrillation properties similar to hAMY while low values indicate no fibrillation.General protocols for acute food intake study

[0085] The effect on cumulative food intake was measured in in male Sprague Dawley rats (6 weeks of age, Taconic, Denmark) following a single dose using a fully automated food intake monitoring system (HM-2; MBRose ApS, Faaborg, Denmark), allowing for advanced synchronous real-time monitoring of food intake behaviour of individual animals. The animals were housed in groups of 4 in a light-, temperature-, and humidity-controlled room (a 12 / 12 LD cycle, lights on at 02:00 AM; 22 ± 2 ° C; 50 % relative humidity). The mice had ad libitum access to regular chow diet (Altromin 1324, Brogaarden A / S, Lynge, Denmark) and domestic quality tap water. Mice arrived at day 7, and a minimum of 5 days of habituation to the system was allowed prior to beginning of the study. During these days, the animals were handled daily to accustom them to the experimental paradigm. On the day of dosing, the animals were randomized into groups according to body weight. Animals were fasted for four hours prior to dosing. Animals were dosed SC 30 or 180 min prior to lights out, and food intake data were collected automatically for a total of 48 h post-dosing with automatic food recordings every 5 min.General protocols for chronic diet-induced obesity (DIO) study

[0086] Sprague-Dawley rats (Taconic, Denmark) arrived at the age of 5 weeks. After one week of acclimation, they were evenly grouped based on their baseline body weight and offered ad libitum access a two-choice diet consisting of chow pellet (Altromin #1324, Brogaarden, Lynge, Denmark) and a high palatable high sugar / fat diet (1:1:1 mixture of Nutella (Ferrero, Alba, Italy), peanut butter (PCD, Rotterdam, Netherlands) and powdered standard chow (Altromin #1324, Brogaarden, Lynge, Denmark); 29.3 % fat, 33.2 % carbohydrate, and 18 % protein) for up to 36 weeks. The rats were pair-housed throughout the study under controlled environmental conditions (12-hour light / dark cycle, lights off at 15:00; 22 ± 1 °C; 50 ± 10% relative humidity). Body weight, food and water intake were monitored daily during the entire period of the study. One day before the experiment, the rats were randomized according to body weight into experimental groups (n = 9-10), which received compound or vehicle SC. QD (once daily) for 28 days.Example 1

[0087] EC 50 values at hAMY3R and hAM1R were measured according to the general protocol above. Percent identity for the hAM 15-52 analogues were calculated in comparison to hAM 15-52 . The results are summarized in Table 2 below. Table 2. Table 2 shows that hAM 15-52 analogues (SEQ ID NO: 3-392) with improved hAMY3R potency compared to hAM 15-52 may be obtained by changing the amino acid K present in hAM 15-52 in position X 11 into R, W or Cit. Table 2 further shows that highly selective (hAMY3R-EC 50 ≤ 250 pM and an hAM1R-EC 50 ≥ 25 nM) hAMY3R hAM 15-52 analogues (SEQ ID NO: 3-360) may be obtained by abolishing or reducing the hAM1R potency using the positions X 4 , X 3 , and / or X 38 . All lipidations at position 1 were performed at the N-terminal and the lipidation in the other positions were performed at an epsilon N in Lys. AMCHC refers to trans-4-(aminomethyl)cyclohexanecarboxylic acid. OEG refers to 8-amino-3,6-dioxaoctanoic acid or 8Ado.ID hAMY3R EC 50 (nM) hAM1R EC 50 (nM) identity (% ADM) ∗< Lipidation Lipidation position 11.2741.113100.0NoneNA20.01>5000.00018.4NoneNA30.006123.54873.7NoneNA40.006>5000.00063.2C20DA-yGlu150.006>5000.00063.2C20DA-yGlu160.006>5000.00063.2C20DA-yGlu-OEG-OEG3670.006>5000.00063.2NoneNA80.006>5000.00044.7NoneNA90.006>5000.00044.7NoneNA100.007551.97068.4NoneNA110.007>5000.00063.2C20DA-yGlu1120.007>5000.00060.5C20DA-yGlu-OEG-OEG27130.007>5000.00063.2NoneNA140.007>5000.00063.2NoneNA150.007>5000.00050.0NoneNA160.008125.46778.9NoneNA170.008294.94271.1NoneNA180.008817.24268.4NoneNA190.008>5000.00028.9NoneNA200.008>5000.00071.1NoneNA210.008>5000.00063.2C20DA-yGlu1220.008>5000.00063.2C20DA-yGlu1230.008>5000.00063.2NoneNA240.008>5000.00063.2NoneNA250.008>5000.00063.2NoneNA260.008>5000.00063.2NoneNA270.008>5000.00063.2NoneNA280.008>5000.00065.8NoneNA290.008>5000.00055.3NoneNA300.008>5000.00063.2NoneNA310.008>5000.00060.5NoneNA320.008>5000.00050.0NoneNA330.008>5000.00052.6NoneNA340.00954.62773.7NoneNA350.009>5000.00063.2C20DA-yGlu1360.009>5000.00063.2C20DA-yGlu-OEG-OEG3370.010>5000.00063.2C20DA-yGlu1380.010>5000.00063.2C20DA-yGlu1390.010>5000.00065.8C20DA-yGlu1400.010>5000.00060.5C20DA-yGlu-OEG-OEG25410.010>5000.00060.5C20DA-yGlu-OEG-OEG26420.010>5000.00063.2NoneNA430.010>5000.00063.2NoneNA440.010>5000.00063.2NoneNA450.01150.47086.8NoneNA460.01178.48086.8NoneNA470.011500.00063.2C20DA-yGlu1480.011760.53963.2C20DA-yGlu1490.0112981.00081.6NoneNA500.011>5000.00065.8C20DA-yGlu1510.011>5000.00063.2C20DA-yGlu1520.011>5000.00063.2NoneNA530.011>5000.00063.2NoneNA540.01246.19481.6NoneNA550.01263.77663.2C20DA-yGlu1560.012103.25073.7NoneNA570.012590.39763.2C20DA-yGlu1580.012>5000.00021.1NoneNA590.012>5000.00026.3NoneNA600.012>5000.00068.4C20DA-yGlu1610.012>5000.00063.2C20DA-yGlu1620.012>5000.00063.2C20DA-yGlu1630.012>5000.00063.2C20DA-yGlu1640.012>5000.00063.2NoneNA650.012>5000.00057.9NoneNA660.012>5000.00060.5NoneNA670.013116.49771.1C20DA-yGlu1680.013254.72268.4C20DA-yGlu1690.013680.33160.5C20DA-yGlu1700.0132981.00084.2NoneNA710.013>5000.00060.5C20DA-yGlu1720.013>5000.00063.2C20DA-yGlu1730.013>5000.00063.2C20DA-yGlu1740.013>5000.00063.2C20DA-yGlu-OEG-OEG14750.013>5000.00065.8NoneNA760.014>5000.00013.2C20DA-yGlu1770.014>5000.00065.8C20DA-yGlu1780.014>5000.00060.5C20DA-yGlu1790.014>5000.00060.5C20DA-yGlu1800.01544.30315.8C20DA-yGlu1810.015138.50084.2NoneNA820.015612.15863.2C20DA-yGlu1830.015815.47363.2C20DA-yGlu1840.0152981.00081.6NoneNA850.015>5000.00063.2C20DA-yGlu1860.015>5000.00060.5C20DA-yGlu1870.015>5000.00060.5C20DA-yGlu1880.015>5000.00060.5C20DA-yGlu1890.015>5000.00060.5C20DA-yGlu-OEG-OEG20900.01658.53081.6NoneNA910.016839.59163.2C20DA-yGlu1920.016932.79560.5C20DA-yGlu1930.016>5000.00028.9NoneNA940.016>5000.00060.5C20DA-yGlu1950.016>5000.00060.5C20DA-yGlu1960.016>5000.00060.5C20DA-yGlu1970.016>5000.00063.2C20DA-yGlu1980.01766.88086.8NoneNA990.017364.19871.1C20DA-yGlu11000.017>5000.00060.5C20DA-yGlu11010.017>5000.00063.2NoneNA1020.018>5000.00071.1NoneNA1030.018>5000.00063.2C20DA-yGlu11040.018>5000.00063.2C20DA-yGlu11050.018>5000.00060.5C20DA-yGlu11060.018>5000.00060.5C20DA-yGlu11070.019646.95060.5C20DA-yGlu11080.019>5000.00060.5C20DA-yGlu11090.0202981.00084.2NoneNA1100.020>5000.00063.2C20DA-yGlu11110.0212981.00089.5NoneNA1120.021>5000.00060.5C20DA-yGlu11130.0222981.00089.5NoneNA1140.022>5000.00063.2NoneNA1150.023129.80084.2NoneNA1160.02448.17089.5NoneNA1170.02451.55578.9NoneNA1180.0242981.00084.2NoneNA1190.024>5000.00065.8C20DA-yGlu11200.025>5000.00057.9C20DA-yGlu11210.0262981.00084.2NoneNA1220.026>5000.00060.5C20DA-yGlu11230.02731.65373.7NoneNA1240.02734.84086.8NoneNA1250.0272981.00086.8NoneNA1260.027>5000.00063.2C20DA-AMCHC-yGlu-OEG-OEG11270.028107.40092.1NoneNA1280.028130.30034.2NoneNA1290.029366.90089.5NoneNA1300.030>5000.00057.9C20DA-yGlu11310.03133.76092.1NoneNA1320.0312981.00089.5NoneNA1330.031>5000.00057.9C20DA-yGlu11340.0322981.00086.8NoneNA1350.03368.40086.8NoneNA1360.033>5000.00060.5C20DA-yGlu-OEG-OEG171370.034>5000.00057.9C20DA-yGlu11380.0352981.00086.8NoneNA1390.0352981.00084.2NoneNA1400.03631.40092.1NoneNA1410.036258.40089.5NoneNA1420.0362981.00084.2NoneNA1430.0382981.00084.2NoneNA1440.038>5000.00057.9C20DA-yGlu11450.0392981.00089.5NoneNA1460.0402981.00092.1NoneNA1470.0403108.87886.8NoneNA1480.040>5000.00026.3NoneNA1490.040>5000.00028.9NoneNA1500.040>5000.00089.5NoneNA1510.04144.92084.2NoneNA1520.041212.70089.5NoneNA1530.041>5000.00060.5C20DA-yGlu11540.0422981.00097.4NoneNA1550.045116.50092.1NoneNA1560.0472981.00086.8NoneNA1570.0472981.00086.8NoneNA1580.0482981.00092.1NoneNA1590.048>5000.00063.2C20DA-yGlu11600.0502981.00086.8NoneNA1610.0512981.00086.8NoneNA1620.051>5000.00031.6NoneNA1630.0532981.00081.6NoneNA1640.054106.20094.7NoneNA1650.0542981.00086.8NoneNA1660.0562981.00084.2NoneNA1670.0562981.00092.1NoneNA1680.062>5000.00092.1NoneNA1690.0632981.00094.7NoneNA1700.0642981.00084.2NoneNA1710.064>5000.00086.8NoneNA1720.0662981.00086.8NoneNA1730.069>5000.00086.8NoneNA1740.071133.30094.7NoneNA1750.0742981.00089.5NoneNA1760.0762981.00086.8NoneNA1770.0772981.00084.2NoneNA1780.0802981.00084.2NoneNA1790.0822981.00089.5NoneNA1800.082>5000.00026.3NoneNA1810.082>5000.00063.2NoneNA1820.085100.10094.7NoneNA1830.0882981.00089.5NoneNA1840.0904495.56486.8NoneNA1850.090>5000.00086.8NoneNA1860.091>5000.00086.8NoneNA1870.0932981.00086.8NoneNA1880.09447.64086.8NoneNA1890.0962981.00089.5NoneNA1900.0972981.00089.5NoneNA1910.0982981.00089.5NoneNA1920.098>5000.00086.8NoneNA1930.099>5000.00084.2NoneNA1940.1123526.54786.8NoneNA1950.112>5000.00060.5NoneNA1960.114>5000.00084.2NoneNA1970.1202981.00086.8NoneNA1980.1223962.08986.8NoneNA1990.1242981.00089.5NoneNA2000.12638.15681.6C20DA-yGlu12010.1264881.06186.8NoneNA2020.126>5000.00086.8NoneNA2030.127>5000.00086.8NoneNA2040.1292981.00086.8NoneNA2050.1303228.70289.5NoneNA2060.130>5000.00086.8NoneNA2070.131>5000.00028.9NoneNA2080.133242.45073.7C20DA-yGlu12090.1361020.00089.5NoneNA2100.137>5000.00063.2NoneNA2110.1512756.64786.8NoneNA2120.153>5000.00089.5NoneNA2130.153>5000.00084.2NoneNA2140.1572981.00086.8NoneNA2150.161>5000.00086.8NoneNA2160.1642981.00086.8NoneNA2170.167>5000.00089.5NoneNA2180.169>5000.00086.8NoneNA2190.1714246.48886.8NoneNA2200.182>5000.00086.8NoneNA2210.184>5000.00063.2C20DA-AMCHC-gGlu-OEG-OEG-K(C20DA-AMCHC-gGlu-OEG-OEG)12220.184>5000.00086.8NoneNA2230.18647.65278.9C20DA-yGlu12240.1862981.00086.8NoneNA2250.186>5000.00065.8C20DA-yGlu12260.186>5000.00086.8NoneNA2270.1874570.81984.2NoneNA2280.205>5000.00086.8NoneNA2290.2072981.00089.5NoneNA2300.208>5000.00089.5NoneNA2310.209>5000.00065.8C20DA-yGlu12320.213>5000.00086.8NoneNA2330.220>5000.00063.2(C20DA-OEG-yGlu-OEG) 2 -K-12340.227>5000.00071.1NoneNA2350.2331416.60086.8NoneNA2360.2342239.41786.8NoneNA2370.2354245.93086.8NoneNA2380.235>5000.00086.8NoneNA2390.236>5000.00086.8NoneNA2400.240>5000.00089.5NoneNA2410.2422291.53486.8NoneNA2420.248>5000.00086.8NoneNA2430,0183336.09863,2NoneNA2440,006>5000.00044,7NoneNA2450,017>5000.00044,7NoneNA2460,010>5000.00063,2NoneNA2470,004>5000.00050.0NoneNA2480,0273587,64671,1NoneNA2490,006>5000.00063,2NoneNA2500,011>5000.00063,2NoneNA2510,006>5000.00063,2NoneNA2520,006>5000.00065,8NoneNA2530,004>5000.00060,5NoneNA2540,006>5000.00063,2NoneNA2550,007>5000.00065,8NoneNA2560,007>5000.00050.0NoneNA2570,0162037,78873,7NoneNA2580,010>5000.00063,2NoneNA2590,0134324,04086,8NoneNA2600,0114163,02386,8NoneNA2610,018>5000.00081,6NoneNA2620,014>5000.00063,2NoneNA2630,0051283,84981,6NoneNA2640,0063469,93673,7NoneNA2650,030>5000.00063,2NoneNA2660,053>5000.00060,5NoneNA2670,0072490,99484,2NoneNA2680,014>5000.00065,8NoneNA2690,005>5000.00081,6NoneNA2700,0031877,71381,6NoneNA2710,01096,61886,8NoneNA2720,009>5000.00071,1NoneNA2730,021>5000.00084,2NoneNA2740,019877,43089,5NoneNA2750,020>5000.00089,5NoneNA2760,056>5000.00063,2NoneNA2770,058>5000.00084,2NoneNA2780,011154,83189,5NoneNA2790,013241,24078,9NoneNA2800,024>5000.00084,2NoneNA2810,034>5000.00084,2NoneNA2820,1433880,50473,7NoneNA2830,035>5000.00086,8NoneNA2840,028590,75092,1NoneNA2850,157>5000.00089,5NoneNA2860,025138,87392,1NoneNA2870,055>5000.00089,5NoneNA2880,030>5000.00086,8NoneNA2890,0053502,99186,8NoneNA2900,047>5000.00084,2NoneNA2910,064>5000.00089,5NoneNA2920,070>5000.00084,2NoneNA2930,063>5000.00089,5NoneNA2940,045>5000.00092,1NoneNA2950,194>5000.00086,8NoneNA2960,392>5000.00089,5NoneNA2970,026>5000.00084,2NoneNA2980,057>5000.00086,8NoneNA2990,110>5000.00086,8NoneNA3000,076>5000.00092,1NoneNA3010,138>5000.00086,8NoneNA3020,0714332,08481,6NoneNA3030,103>5000.00086,8NoneNA3040,139>5000.00084,2NoneNA3050,085>5000.00092,1NoneNA3060,003>5000.00044,7NoneNA3070,010>5000.00044,7NoneNA3080,014>5000.00063,2NoneNA3090,025>5000.00063,2NoneNA3100,003>5000.00050.0NoneNA3110,060>5000.00078,9NoneNA3120,012>5000.00071,1NoneNA3130,004>5000.00063,2NoneNA3140,008>5000.00063,2NoneNA3150,012>5000.00063,2NoneNA3160,012>5000.00065,8NoneNA3170,004>5000.00060,5NoneNA3180,009>5000.00063,2NoneNA3190,018>5000.00065,8NoneNA3200,009>5000.00063,2NoneNA3210,317>5000.00063,2NoneNA3220,0141390,17886,8NoneNA3230,014>5000.00081,6NoneNA3240,028>5000.00063,2NoneNA3250,025>5000.00063,2NoneNA3260,053>5000.00081,6NoneNA3270,090>5000.00073,7NoneNA3280,010>5000.00063,2NoneNA3290,032>5000.00063,2NoneNA3300,008>5000.00065,8NoneNA3310,0063566,75484,2NoneNA3320,007>5000.00081,6NoneNA3330,236107,47163,2NoneNA3340,013>5000.00071,1NoneNA3350,015>5000.00084,2NoneNA3360,029738,47889,5NoneNA3370,0191583,67789,5NoneNA3380,089>5000.00063,2NoneNA3390,047>5000.00084,2NoneNA3400,022671,05889,5NoneNA3410,029>5000.00084,2NoneNA3420,054>5000.00084,2NoneNA3430,017117,36386,8NoneNA3440,121>5000.00089,5NoneNA3450,051113,11692,1NoneNA3460,085>5000.00086,8NoneNA3470,068>5000.00084,2NoneNA3480,041>5000.00084,2NoneNA3490,046>5000.00084,2NoneNA3500,0441180,47092,1NoneNA3510,943>5000.00086,8NoneNA3520,0401116,54184,2NoneNA3530,0272918,79189,5NoneNA3540,091>5000.00086,8NoneNA3550,140>5000.00086,8NoneNA3560,079>5000.00086,8NoneNA3570,115>5000.00086,8NoneNA3580,071>5000.00081,6NoneNA3590,145>5000.00086,8NoneNA3600,165>5000.00084,2NoneNA3610.0060.91376.3NoneNA3620.00718.15086.8NoneNA3630.0101.77171.1NoneNA3640.01013.58584.2NoneNA3650.0121.21186.8NoneNA3660.01312.09089.5NoneNA3670.0140.56389.5NoneNA3680.0150.99789.5NoneNA3690.0172.47784.2NoneNA3700.01710.30473.7C20DA-yGlu13710.0193.02992.1NoneNA3720.0203.94184.2C20DA-yGlu13730.0207.71286.8NoneNA3740.02112.00089.5NoneNA3750.0220.40392.1NoneNA3760.02212.02281.6NoneNA3770.0240.24381.6NoneNA3780.0270.89376.3C20DA-yGlu13790.0290.45889.5NoneNA3800.0310.16286.8C20DA-yGlu13810.0327.60886.8NoneNA3820.0345.12492.1NoneNA3830.0345.93592.1NoneNA3840.0361.09092.1NoneNA3850.0367.00389.5NoneNA3860.0397.35478.9C20DA-yGlu13870.0400.99294.7NoneNA3880.0434.00684.2C20DA-yGlu13890.0451.03971.1C20DA-yGlu13900.0480.88986.8C20DA-yGlu13910.0680.66592.1NoneNA3920.1135.03484.2C20DA-yGlu1 Example 2

[0088] Fibrillation was determined according to the general protocol for determination of physical stability of peptide analogues. The data for selected hAM 15-52 analogues are summarized in Table 3 below. Table 3. Table 3 shows that the hAM 15-52 analogues according to the invention maintain the good fibrillation properties from hAM 15-52 . Most of the fibrillation data are comparable to and some even better than hAM 15-52 (SEQ ID NO: 1) at pH 7 (ThT signal of 2.6 %). All of the fibrillation data are far superior compared to the reference hAMY 1-37 (SEQ ID NO: 2) (ThT signal on 100 %).ID Max ThT signal (%) identity (% ADM) ∗< 12.600100.0210018.472.80063.2135.00063.2143.10063.2209.10071.1234.20063.2246.80063.22533.40063.2260.00063.22721.8063.2288.20065.8299.30055.3325.30050.0426.30063.2431.20063.2453.00086.8465.80086.8493.20081.6527.60063.2642.70063.2651.00057.9708.80084.2814.00084.28414.20081.6902.00081.69812.60086.81099.30084.211314.70089.51142.20063.21152.50084.211611.70089.51187.40084.21211.80084.21244.80086.81257.20086.81291.40089.51316.80092.11325.20089.513417.40086.81353.30086.81386.80086.81394.80084.21403.10092.11413.20089.51432.20084.21452.00089.51463.60092.11475.10089.215018.20091.91521.90089.515615.30086.81586.30092.116021.50086.816114.30086.81632.00081.616424.30094.71654.80086.81662.00084.21676.60092.116812.50094.61692.70094.71701.80084.21715.00089.217213.90086.81734.40089.21753.20089.51766.80086.81818.80063.218215.60094.71840.80089.21852.80089.21860.60089.21871.60086.81896.00089.519012.90089.51916.30089.51924.10089.21932.70086.51946.00089.21956.10060.51960.50086.51975.60086.81984.20089.21998.90089.52011.00089.220212.60089.22031.20089.22042.20086.82057.80091.92061.50089.22101.40063.22115.20089.22121.90091.92136.50086.52142.70086.82150.60089.221713.80091.92184.50089.22190.70089.22201.10089.222211.70089.22242.00086.822627.00089.22278.50086.522819.60089.22300.60091.92321.20089.22350.80089.22360.60089.22370.30089.22380.60089.22390.20089.22403.70091.92410.90089.22420.80089.22438.55063.22444.10044.72453.35044.72464.20063.22471.35050.024813.60071.124915.60063.22506.40063.225110.60063.225227.55063.22539.60060.525412.05063.225525.00065.825618.65050.025720.95073.725832.70063.225913.45086.826034.10086.826122.65081.62629.55063.22631.05081.62641.35073.726526.55063.226620.90060.526730.90084.22688.85065.826934.50081.627025.45081.627110.85086.827216.75071.127329.00084.227426.80089.527519.85089.52768.95063.227721.60084.227813.35089.527913.45078.928026.45084.228122.65084.22824.70073.728317.10086.828421.30092.128531.20089.52862.90092.128710.65089.528817.45086.828921.15086.829017.30084.229119.20089.529219.70084.229324.20089.529416.85092.129519.45086.829626.05089.529727.50084.229831.25086.829926.55086.830017.85092.130116.95086.830224.25081.630318.15086.830422.35084.230524.60092.13063.00044.730711.20044.730814.50063.23094.25063.23103.50050.03116.45078.931217.00071.131315.05063.231411.90063.231517.15086.831620.45089.53175.55086.831814.90089.53193.90092.132011.10089.532121.70094.732218.40092.132327.50081.632411.85063.232510.10063.23262.35081.63273.60073.732833.25063.23296.35063.233014.60065.833121.15084.23322.10081.633325.70063.233410.15071.13351.60084.233618.70089.533716.80089.53382.95063.233920.05084.23401.70089.534120.50084.234219.80084.234324.85086.834413.35089.53453.25092.13461.20086.834715.50084.23482.75084.234915.25084.235015.70092.13517.20086.835225.80084.235330.30089.535417.40086.835525.35086.83561.25086.835719.75086.835824.35081.635920.35086.836026.35084.23628.90086.83663.30089.537318.00086.83744.80089.53839.60092.13858.80089.538714.30094.73913.70092.1 Example 3

[0089] The effect of selected hAM 15-52 analogues on acute food intake in lean rats were determined. The results are summarized in Table 4 below. Table 4. Table 4 shows that hAM 15-52 analogues have potent anorectic effects for up to 48 hours following a single SC injection in lean healthy rats (n=7-8), thereby providing proof-of-principle. Dunnett's test one-factor linear model***: P < 0.001 compared to Vehicle (three independent runs were conducted).ID Dose Average cumulative food intake (g) % to vehicle Significance level to Veh NAVehicle44.4100NA410 nmol / kg26.359.23***430 nmol / kg13.730.86***4610 nmol / kg23.352.48***4630 nmol / kg10.724.10***6010 nmol / kg30.969.59***6030 nmol / kg20.345.72***NAVehicle40.7100NA7810 nmol / kg13.733.66***6910 nmol / kg7.1317.52***7010 nmol / kg24.961.18***8510 nmol / kg18.645.70***12410 nmol / kg24.259.46***9310 nmol / kg19.146.93***NAVehicle41.9100NA3610 nmol / kg23.355.61***610 nmol / kg27.565.63*** Example 4

[0090] The Effect of chronic treatment of selected hAM 15-52 analogues according to the invention in diet-induced obese (DIO) rats were determined. The results are summarized in Table 5 below. Table 5. Table 5 shows that daily SC injections of SEQ ID NO: 46 or 78 cause a significant reduction in body weight in diet-induced obese rats (n=10) following a 28-day treatment period, thereby providing proof-of-concept. Dunnett's test one-factor linear model ∗∗∗< : P < 0.001 compared to Vehicle (two independent runs were conducted).ID Dose Average body weight day 28 (gram) % to vehicle Significance level to Veh NAVehicle716100NA461 nmol / kg67494.13***463 nmol / kg62086.59***4610 nmol / kg61886.31***NAVehicle776100NA7810 nmol / kg68788.53*** Example 5

[0091] Deeper investigation of representative amino acids in the position X 4 , X 11 , X 17 , X 3 , and X 38 and the corresponding change in potency. The results are summarized in Table 6 below. Table 6. Table 6 shows whether the amino acid change causes increase (I) in hAMY3R potency, unchanged potency (U), or decreased potency (D) compared to the reference (R) amino acid. N / A Not available.Amino acid Position 4 Position 11 Position 17 Position 37 Position 38 Y UDRIIW IUIIIT RDURUQ DDUUUP DDDIUM IDUUII IDUUUH DDIIUF IDUIIE DDDUUA UDUUUR DRUUUC IDDUN / AK DDUUUHyp N / AN / AN / AN / ARCit N / AUN / AN / AN / A

Claims

1. An hAM15-52 analogue or a pharmaceutically acceptable salt thereof comprising 38 amino acids (X1-X38) with an hAMY3R-EC50 ≤ 250 pM and an hAM1R-EC50 ≥ 25 nM, wherein the amino acid in position X4 is selected as F, Y, W, T, M, I, A, or C; X37 is selected as G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X38 is selected as Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; X11 is R, W, or Cit and wherein at least one of the positions X4, X37 or X38 is not the amino acid present in hAM15-52 (SEQ ID NO: 1) in said position and further wherein the hAM15-52 analogue has at least 50 % identity to hAM15-52 (SEQ ID NO: 1).

2. An hAM15-52 analogue or a pharmaceutically acceptable salt thereof according to claim 1, wherein the hAM15-52 analogue has at least 60 % identity to hAM15-52, such as at least 70 % identity to hAM15-52, preferably at least 80 % identity to hAM15-52 (SEQ ID NO: 1).

3. An hAM15-52 analogue or a pharmaceutically acceptable salt thereof according to claim 1, comprising the amino acid sequence of formula (I): wherein X11 is selected from R, W, or Cit; X17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X4 is selected from F, Y, W, T, M, I, A, or C; X37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X4, X37 or X38 is not the amino acid present in hAM15-52 (SEQ ID NO: 1) in said position; or a derivative thereof with at least 50 % identity to hAM15-52, such as at least 55 % identity to hAM15-52, such as at least 60 % identity to hAM15-52, such as at least 65 % identity to hAM15-52, preferably at least 70 % identity to hAM15-52, such as at least 75 % identity to hAM15-52,, more preferably at least 80 % identity to hAM15-52, such as at least 90 % identity to hAM15-52, preferably at least 95 % identity to hAM15-52, wherein, in the derivative, X11 is selected from R, W, or Cit; X17 is selected as Y, W, T, Q, M, I, H, F, A, R, or K; X4 is selected from F, Y, W, T, M, I, A or C; X37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X4, X37 or X38 is not the amino acid present in hAM15-52 (SEQ ID NO: 1) in said position.

4. An hAM15-52 analogue or a pharmaceutically acceptable salt thereof according to claim 1 with the sequence SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360 or a derivative of any thereof with at least 90 % identity to any one of SEQ ID NO: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, preferably at least 95 % identity to any one of SEQ ID NO: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, most preferably at least 97 % identity to anyone of SEQ ID NOs: 3-57, 60-75, 77-79, 81-92, 94-127, 129-147, 150-161, 163-179, 181-206, 208-243, 246-305, or 308-360, with the proviso that in the derivative X11 is selected from R, W, or Cit; X17 is selected as Y, W, T, Q, M, I, H, F, A, R or K; X4 is selected from F, Y, W, T, M, I, A, or C; X37 is selected from G, Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X38 is selected from Hyp, Y, W, T, Q, P, M, I, H, F, E, A, R, or K; and further wherein at least one of the positions X4, X37 or X38 is not the amino acid present in hAM15-52 (SEQ ID NO: 1) in said position.

5. An hAM15-52 analogue or a derivative thereof according to any of the preceding claims, wherein at least two of the positions X4, X37, and X38 is not the amino acid present in hAM15-52 (SEQ ID NO: 1) in said position.

6. An hAM15-52 analogue or a derivative thereof according to any of the preceding claims, wherein all of the positions X4, X37 and X38 are selected as follows: X4 is selected from Y, W, T, M, I, A, or C; X37 is selected from Y, S, W, T, Q, P, M, I, H, F, E, A, R, C, or K; X38 is selected from Hyp, W, T, Q, P, M, I, H, F, E, A, R, or K;7. An hAM15-52 analogue or a derivative thereof according to any of the preceding claims, wherein X4 is selected as F, W, M, I, or C.

8. An hAM15-52 analogue or a derivative thereof according to any of the preceding claims, wherein X37 is selected as G, Y, W, P, H, or F.

9. An hAM15-52 analogue or a derivative thereof according to any of the preceding claims, wherein X38 is selected as Hyp, Y, W, M, or F.

10. An hAM15-52 analogue or a derivative thereof according to any of the preceding claims, wherein X17 is selected as W, or H.

11. An hAM15-52 analogue or a derivative thereof according to any of the preceding claims, wherein X11 is R.

12. An hAM15-52 analogue or a derivative thereof according to any of claims 1-11 for use as a medicament.

13. A pharmaceutical composition comprising an hAM15-52 analogue or a derivative thereof according to claims 1-11, a pharmaceutically acceptable carrier and optionally one or more excipient(s).