Secretin-derived GLP1R selective agonists

Abstract

The present invention relates to selective GLP1R agonists. In particular, the present invention relates to the discovery that secretin (SCT) can be converted into a high-potency and high-selectivity GLP1R agonist.

Description

TECHNICAL FIELD

[0001] The present invention relates to selective GLP1R agonists. In particular, the present invention relates to the discovery that secretin (SCT) can be converted into a highly potent and selective GLP1R agonist. The present invention further relates to GLP1R agonists for use as a medicament, in particular for the treatment of obesity and / or diabetes. BACKGROUND

[0002] Obesity is a medical condition in which excess body fat accumulates to the extent that it has a negative impact on health. It affects a large number of people globally and the incidence is rapidly rising in parts of the world. The World Health Organization (WHO) estimates that in 2016, approximately 650 million people worldwide suffered from obesity. Obesity is defined as a body mass index (BMI) of over 30. Obesity is considered a major risk factor for suffering from a number of medical conditions, such as cardiovascular disease (e.g. high blood pressure, atherosclerosis, heart attack or stroke), NASH, musculoskeletal disease, certain types of cancer, depression and type 2 diabetes, and is therefore detrimental to human health.

[0003] Diabetes is a group of metabolic disorders characterized by high blood sugar levels. As of 2019, the International Diabetes Federation estimates that 463 million people worldwide have diabetes, of which approximately half have been diagnosed. Diabetes is divided into two types, i.e. type 1 diabetes and type 2 diabetes. Type 1 diabetes is due to an autoimmune response that leads to damage to beta cells, so that the pancreas cannot produce enough insulin. On the other hand, type 2 diabetes is a condition in which insulin resistance is initially, i.e. cells do not respond normally to insulin, and can also lead to insulin deficiency as the disease progresses.

[0004] The secretin family of hormones is a group of short chain brain-gut peptides named after the first hormone discovered in the group. The family members include the following peptides: secretin (SCT), pituitary adenylate cyclase-activating peptide (PACAP), vasoactive intestinal peptide (VIP), growth hormone-releasing hormone (GHRH), glucagon (GCG), glucagon-like peptide-1 (GLP-1), glucagon-like peptide-2 (GLP-2), glucose-dependent insulinotropic polypeptide (GIP), corticotropin-releasing hormone (CRH), and calcitonin (CALC). These peptide hormones exert their effects by activating B- or group 2 G protein-coupled receptors.

[0005] SCT is a hormone produced by enteroendocrine S cells in intestinal glands located in the duodenum and jejunum. Secretin (SCT) regulates water homeostasis throughout the body and influences the duodenal environment by modulating secretions from the stomach, pancreas, and liver. SCT inhibits gastric acid secretion and gastric emptying and, in response to ingested food, stimulates exocrine pancreatic secretion of water and bicarbonate, thereby ensuring protection of the intestinal mucosa and ensuring an optimal chemical environment for digestive enzymes. Secretin is initially synthesized as a 120 amino acid precursor protein, termed prosecretin, which contains an N-terminal signal peptide, a spacer, secretin itself (residues 28-54), and a 72 amino acid C-terminal peptide. Mature secretin is a linear peptide hormone consisting of 27 amino acids with the sequence HSDGTFTSELSRLREGARLQRLLQGLV.

[0006] Glucagon-like peptide-1 (GLP-1) is produced by enteroendocrine L cells in the gut and, in response to ingested food, is secreted into the hepatic portal system. GLP-1 has several physiological properties that make it and its functional analogs an object of intensive research for the treatment of diabetes and obesity. In the stomach, GLP-1 inhibits gastric emptying, acid secretion, and motility, effects that collectively reduce appetite. Likewise, GLP1R is highly enriched in the circumventricular organs (CVOs) and hypothalamic nuclei involved in the regulation of energy balance, and GLP1R activation in these regions has been shown to reduce food intake. In addition, GLP-1 acts as an incretin hormone, stimulating the release of insulin and inhibiting the secretion of glucagon by the pancreas in response to ingested food. These effects have been clinically exploited through the development of GLP1R agonists, such as Liraglutide, for the treatment of diabetes and obesity.

[0007] Evolutionarily related peptides of the secretin family have some sequence homology. For example, secretin shares 11 amino acids with GLP-1, as shown below:

[0008] Secretin: HSDGTFTSELSRLREGARLQRLLQGLV

[0009] GLP-1: HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR

[0010] Based on their sequence homology, secretin has previously been used as a template to design a high potency and long acting SCTR / GLP1R co-agonist (GUB06-046) by introducing GLP-1 residues (Witteloostuijn, 2017). The amino acid sequence of SCTR / GLP1R co-agonist GUB06-046 (SEQ ID NO: 216) is H-Aib-E-G-T-F-T-S-D-L-S-R-L-L-E-G-A-A-L-Q-R-F-I-Q-W-L-V, the EC 50 for SCTR is 1.3 nM and the EC 50 for GLP1R is 0.12 nM.

[0011] Despite the large number of GLP-1 analogs, liraglutide and semaglutide are still the only GLP-1 analogs approved for the treatment of obesity. Therefore, there is a need for new GLP1R agonists for the treatment of obesity and / or diabetes, in particular GLP1R agonists based on a different chemical scaffold. The present invention aims to solve the problem of providing a high potency and highly selective GLP1R agonist based on the secretin scaffold. SUMMARY

[0012] The present invention relates to novel GLP1R agonists based on the SCT scaffold. The inventors found that SCT can be converted into a GLP1R agonist with high potency and high selectivity. The present invention is recited in the claims. BRIEF DESCRIPTION OF DRAWINGS

[0013] Embodiments of the present invention will be described in more detail below with reference to the accompanying drawings.

[0014] Figure 1A The effect on hGLP1R potency after introducing GLP-1 amino acid residues present at position X 2 , X 3 , X 9 , X 10 , X 12 , X 13 , X 14 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 or X 25 is shown below. A positive average SHAP value indicates an improvement in hGLP1R potency, a negative average SHAP value indicates a decrease in hGLP1R potency.

[0015] Figure 1B shows the effect on hSCTR potency after introducing GLP-1 amino acid residues present at X 2 , X 3 , X 9 , X 10 , X 12 , X 13 , X 14 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 or X 25 into the corresponding site of the SCT scaffold. Positive average SHAP values indicate improved hSCTR potency, and negative average SHAP values indicate decreased hSCTR potency.

[0016] Figure 2 shows the normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at X 2 .

[0017] Figure 3A shows the normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at X 3 .

[0018] Figure 3B shows the ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e., selectivity) for different amino acids tested at X 3 .

[0019] Figure 4A shows the normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at X 9 .

[0020] Figure 4B shows the ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e., selectivity) for different amino acids tested at X 9 .

[0021] Figure 5A shows the normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at X 10Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X

[0022] Figure 5B Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 10 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0023] Figure 6A Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 14 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0024] Figure 6B Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 14 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0025] Figure 7 Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 18 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0026] Figure 8A Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 19 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0027] Figure 8B Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 19 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0028] Figure 9 Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 22 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0029] Figure 10A Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 12 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0030] Figure 10B Normalized pEC50 values for hSCTR and hGLP1R for different amino acids tested at position X 12 Ratio of normalized pEC50 values for hSCTR and hGLP1R (i.e. selectivity) for different amino acids tested at position X

[0031] Figure 11A Normalized pEC50 values for hSCTR and hGLP1R are shown for different amino acids tested at X 25

[0032] Figure 11B Ratio of normalized pEC50 for hSCTR to hGLP1R (i.e., selectivity) is shown for different amino acids tested at X 25

[0033] Figure 12 Effect on hGLP1R pEC50 is shown after introducing different lipidated lysine (K) residues at X 1 -X 30

[0034] Figure 13A Effect on hGLP1R potency is shown after introducing glutamic acid (E) residues at the indicated positions. Positive SHAP values indicate improvement in hGLP1R potency, and negative SHAP values indicate decrease in hGLP1R potency.

[0035] Figure 14A Effect on fibril formation at pH 7 is shown after introducing the indicated amino acids at X 3 , X 10 , X 12 , X 18 , X 19 , and X 21 Negative SHAP values indicate that the amino acid minimizes fibril formation, and positive SHAP values indicate that the amino acid increases fibril formation.

[0036] Figure 14B Effect on hGLP1R potency is shown after introducing the indicated amino acids at X 3 , X 10 , X 12 , X 18 , X 19 , and X 21 Positive SHAP values indicate improvement in hGLP1R potency, and negative SHAP values indicate decrease in hGLP1R potency.

[0037] Figure 15A Effect on hGLP1R potency is shown for extended GLP1R agonists (i.e., having 28 to 30 amino acids). Positive average SHAP values indicate improvement in hGLP1R potency, and negative average SHAP values indicate decrease in hGLP1R potency.

[0038] Figure 16A ​​​The effect of SEQ ID NO: 203 compared to vehicle on cumulative food intake is shown.

[0039] Figure 16B The percentage of cumulative food intake after 24 hours for SEQ ID NO: 203 compared to vehicle (vehicle set to 100%) is shown.

[0040] Figure 17A The effect of SEQ ID NO: 209 compared to vehicle on cumulative food intake is shown.

[0041] Figure 17B The percentage of cumulative food intake after 24 hours for SEQ ID NO: 209 compared to vehicle (vehicle set to 100%) is shown.

[0042] Definitions

[0043] The GLP1R agonist of the application is preferably amidated at the C-terminus (-CONH2), similar to the natural peptides secretin (SCT) and glucagon-like peptide-1 (GLP-1). However, the GLP1R agonist of the application can also have a free carboxylic acid (-COOH) or other post-translational modification, such as a methyl ester (-COOMe). In the most preferred embodiment of the application, the GLP1R agonist is amidated at the C-terminus, similar to the peptides exemplified herein. The GLP1R agonist of the application can have a free amine (-NH2), an N-acylation (-NHCOR), an N-methylation (-NHCH3 or -N(CH3)2) or a deaminated form at the N-terminus. Most preferably, the GLP1R agonist of the application has a free amine (i.e. an amino group (-NH2)) at the N-terminus, similar to the peptides exemplified herein.

[0044] In the present context, lipidation means that a lipid is covalently linked to the GLP1R agonist of the application, optionally via a linker / spacer. The lipid can be C18DA (octadecanedioic acid), C20DA (eicosanedioic acid), optionally linked via a linker / spacer consisting of one or more commonly used covalent linking units. Commonly used linkers / spacers can include, but are not limited to, [yE], [OEG] or [AHX] as shown below, for example.

[0045]

[0046] Lipidation is typically performed to improve the pharmacokinetic profile of the polypeptide, such as to improve metabolic stability, reduce enzymatic degradation, reduce excretion and metabolism, in general resulting in an extended half-life (t½) in vivo. Depending on the desired half-life, the GLP1R agonist of the application can be lipidated or non-lipidated. The polypeptide can be lipidated, for example at a lysine (K) residue exemplified herein. Preferably, the lipidation is at X 14 or X25 Lysine residue (K) at position. The lipid (and linker) is selected from the list consisting of C14 (tetradecanoic acid), C16 (hexadecanoic acid), C18DA, C20DA, C18DA[γΕ]-, C18DA[γΕ][γΕ]-, C18DA[γΕ][OEG]-, C18DA[γΕ][OEG][OEG]-, C18DA[γΕ][γΕ][OEG][OEG]-, C18DA[γΕ][AHX]-, C18DA[γΕ][γΕ][AHX]-, C20DA[γΕ]-, C20DA[γΕ][γΕ]-, C20DA[γΕ][OEG]-, C20DA[γΕ][OEG][OEG]-, C20DA[γΕ][γΕ][OEG][OEG]-, C20DA[γΕ][AHX]-, or C20DA[γΕ][γΕ][AHX]-. Preferably, the lipid (and linker) is selected from the list consisting of C18DA[γΕ]-, C18DA[γΕ][γΕ]-, C18DA[γΕ][OEG]-, C18DA[γΕ][OEG][OEG]-, C18DA[γΕ][γΕ][OEG][OEG]-, C20DA[γΕ]-, C20DA[γΕ][γΕ]-, C20DA[γΕ][OEG]-, C20DA[γΕ][OEG][OEG]-, or C20DA[γΕ][γΕ][OEG][OEG]-. Most preferably, the lipidation is C20DA[γΕ][OEG][OEG]-.

[0047] Unless otherwise indicated, the amino acids herein refer to the natural amino acids (i.e., the L form of the amino acids). The abbreviation Aib refers to 2-aminoisobutyric acid.

[0048] EC 50 Values are used as a measure of the potency of the Exendin Receptor (SCTR) and GLP-1 Receptor (GLP1R) agonist. EC 50 Values refer to a measure of the concentration of a compound required to achieve half of its maximum activity in a particular assay. pEC 50 is calculated as -log(EC 50 ), where EC 50 is reported in M. If EC 50 equals 1 micromolar (10 -6 Molar), then pEC 50 is 6.

[0049] The GLP1R agonist of the present invention can be in the form of a pharmaceutically acceptable salt and / or solvate. Thus, pharmaceutically acceptable salts are intended to include, but are not limited to, any salt of the peptide formulation commonly used. Such salts include acid addition salts and basic salts, and examples of which can be found in, for example, Remington’s Pharmaceutical Sciences 17thEdition.

[0050] In the present context, a selective GLP1R agonist is to be understood as an agonist having a SCTR EC 50 / GLP1R EC 50 ratio of at least 100, preferably a SCTR EC 50 / GLP1R EC 50 ratio of at least 500, more preferably a SCTR EC 50 / GLP1R EC 50 ratio of at least 1000, most preferably a SCTR EC 50 / GLP1R EC 50 ratio of at least 10000. SCTR EC 50 / GLP1R EC 50 The ratio can also interchangeably be referred to as selectivity.

[0051] In the present context, homology to SCT is calculated by aligning each peptide to SCT and determining the number of identical amino acid residues. As a non-limiting example, SEQ ID NO: 1 has 16 amino acid residues identical to SCT when aligned as shown below, and thus the homology is (16 / 27 x 100) = 59%.

[0052] SEQ ID NO. 1 : H-X-E-G-T-F-T-S-D-V-S-R-L-K-E-E-A-A-X-Q-R-F-L-E-H-L-V (X is Aib)

[0053] Exendin: H-S-D-G-T-F-T-S-E-L-S-R-L-R-E-G-A-R-L-Q-R-L-L-Q-G-L-V

[0054] As another non-limiting example, SEQ ID NO: 204 has 18 amino acid residues identical to SCT when aligned as shown below, and thus the homology is (18 / 27 x 100) = 67%.

[0055] SEQ ID NO. 204: H-X-E-G-T-F-T-S-D-V-S-R-L-K-E-G-A-A-E-Q-R-F-L-Q-H-L-V-E-A-E (X is Aib)

[0056] Secretin: H-S-D-G-T-F-T-S-E-L-S-R-L-R-E-G-A-R-L-Q-R-L-L-Q-G-L-V

[0057] As shown herein, these peptides have at least 50% homology to SCT. Since at least X 2 , X 3 , X 9 , X 10 , X 14 , X 18 and X 22 amino acids of SCT are different, the maximum SCT identity is (27-7) / 27 x 100 = 74%. The peptides exemplified herein with the lowest SCT identity have 15 amino acids identical to SCT when aligned. Thus, in the most preferred embodiments, these peptides have at least 55% homology to SCT. DETAILED DESCRIPTION

[0058] The present invention relates to novel GLP1R agonists based on the SCT backbone. The inventors have found that SCT can be transformed into a high potency and selective GLP1R agonist. The inventors initially constructed and tested a peptide library in which one or more amino acids in secretin were replaced with the corresponding amino acid present in GLP-1 (see Example 1). Based on this peptide library, the inventors found that X 2 , X 9 , X 18 and X 22 positions in SCT are critical for increasing GLP1R potency of the SCT backbone (see Figure 1A ). Likewise, the inventors found that X 3 , X 9 , X 10 , X 14 and X 19 positions in SCT are critical for decreasing SCTR potency of the secretin backbone for selectivity. Based on these findings, the inventors further found that the non-GLP-1 amino acids in X 2 , X 3 , X 9 , X 10 , X 14 , X 18 , X 19 and X 22 positions of SCT are critical for transforming the secretin backbone into a high potency and selective GLP1R agonist (see Example 2).

[0059] In summary, the inventors have found that in order to provide a selective GLP1R agonist, X2 The amino acid S at position S should be replaced with A, Aib, or G; X in SCT 3 The amino acid D at position D should be replaced with I, H, L, W, V, Y, E, or Q; X in SCT 9 The amino acid E at position X should be replaced with D; 10 The amino acid L at position X should be replaced with P, Aib, V, I, Y, or W; 14 The amino acid R at position X18 of SCT should be replaced with F, Y, W, L, H, Aib, or K; the amino acid R at position X18 of SCT should be replaced with A, Aib, or L; the amino acid R at position X18 of SCT should be replaced with A, Aib, or L; 19 The amino acid at position X should be selected from L, N, G, S, T, Aib, A, E, Q, I, or V; and the X of SCT... 22 The amino acid L at position L should be replaced with F, W, or Y.

[0060] The SCTR / GLP1R co-agonist GUB06-046 (SEQ ID NO: 216) has the amino acid sequence H-Aib-EGTFTSDLSRLLEGAALQRFI-QWLV, which contains some identified amino acid residues that can improve the GLP1R efficacy of the SCT backbone and reduce its SCTR efficacy. Therefore, this invention, starting with GUB06-046, relates to a key discovery: through X 10 Modification at the X site can significantly improve GLP1R selectivity. Table 3 shows a comparison between the peptide GUB06-046 of the present invention and the prior art. As can be seen from Table 3, modifying GUB06-046 X 10 The amino acid L at position L is replaced with P, Aib, V, I, Y, or W, resulting in a significant improvement in the peptide's selectivity for GLP1R relative to SCTR (i.e., a selectivity ratio of at least 100) while maintaining sufficient GLP1R efficacy.

[0061] Key positions in the SCT scaffold for providing high potency and selective GLP1R agonists

[0062] X 2 Bit: X 2 The position was found to be crucial for hGLP1R efficacy (see Example 1). Deep mutation scanning revealed that X 2 The amino acids Aib, A, and G at position A produce the highest hGLP1R efficiency (see [link]). Figure 2 (The box in A). Therefore, the inventors discovered that in order to improve the hGLP1R performance of the SCT skeleton, X 2 The position should be selected as A, Aib, or G. Most preferably, X... 2 The bit selection is Aib.

[0063] X 3 Bit: X3 The position was found to be crucial for reducing hSCTR efficacy to provide a selective GLP1R agonist (see Example 1). Deep mutation scanning revealed that, except for amino acid E of GLP-1 (Example 1), the position associated with SCT X... 3 Compared to amino acid D, non-GLP-1 amino acids I, H, L, W, V, Y, G, R, Aib, P, K, A, F, Q, T, N, and S were found to reduce hSCTR efficiency (see [link to relevant documentation]). Figure 3A To achieve the highest selectivity of GLP1R relative to hSCTR, in X 3 Preferably, the reduction in hSCTR performance should be accompanied by the smallest possible sacrifice in hGLP1R performance. Figure 3B In this context, amino acids are classified according to selectivity (i.e., hSCTR EC). 50 / hGLP1R EC 50 Sort. EC50 of a given amino acid. 50 The higher the ratio, the higher the selectivity for GLP1R. Therefore, when using X... 3 When maximizing bit selectivity, preferably, X 3 Selected from I, H, L, W, V, Y, G, R, Aib, P, K, or A (EC) 50 Ratio = 0.5), more preferably, X 3 Selected from I, H, L, W, V, Y, G, or R (EC) 50 (Ratio = 0.75), or even more preferably, X 3 Selected from I, H, L, W, V or Y (EC) 50 (Ratio = 1.0), and more preferably, X 3 Selected from I, H or L (EC) 50 Ratio = 1.5), most preferably, X 3 The choice is I. Since utilizing multiple sites reduces hSCTR efficiency, X is preferable even if it's not optimal from a selectivity perspective. 3 The position can also include amino acids that preferentially have high hGLP1R efficiency. Therefore, by Figure 3A It can be seen that X 3 The bits can be chosen as E, Q, L, and I to maintain maximum hGLP1R performance, while hSCTR performance is reduced to varying degrees (see [link]). Figure 3A (The box in the middle). Therefore, in the preferred embodiment, X 3 The bit can be selected as I, H, L, W, V, Y, E, or Q to improve selectivity. In a more preferred embodiment, X 3 The bit is selected as I, H, L, E, or Q to improve selectivity. In the most preferred embodiment, X 3 The bit selection is E.

[0064] X 9 Position: X 9 Position X was found to be critical for increasing hGLP1R potency and decreasing hSCTR potency to provide a selective GLP1R agonist (see Example 1). A deep mutational scan in Example 2 showed that the GLP-1 amino acid D at position X 9 was more advantageous in improving hGLP1R potency and decreasing hSCTR potency than all tested amino acids (see Example 2, Figure 4A and Figure 4B ). Thus, in the most preferred embodiment, X 9 is selected as D to increase hGLP1R potency and decrease hSCTR potency of the SCT scaffold.

[0065] X 10 Position: X 10 Position X was found to be critical for decreasing hSCTR potency to provide a selective GLP1R agonist (see Example 1). A deep mutational scan showed that the non-GLP-1 amino acids P, Aib, V, T, A, G, S, I, D, E, N, K, H, Q, Y, W, R, F were found to decrease hSCTR potency compared to the amino acid L found at position X 10 of SCT (see Example 2, Figure 5A ). To achieve the highest selectivity of GLP1R over hSCTR, at position X 10 , preferably, the decrease in hSCTR potency should be accompanied by as little sacrifice in hGLP1R potency as possible. In Figure 5B , the amino acids are ranked by selectivity (i.e., hSCTR EC 50 / hGLP1R EC 50 ). The higher the EC 50 ratio of a given amino acid, the higher the selectivity for GLP1R. Thus, when maximizing selectivity using position X 10 , preferably, X 10 is selected from P, Aib, V, T, A, G, S, I, D, E, N, or K (EC 50 ratio threshold = 0.3), more preferably, X 10 is selected from P, Aib, V, T, A, G, S, or I (EC 50 ratio threshold = 0.4), even more preferably, X 10 is selected from P, Aib, V, T, or A, most preferably, X 10 is selected from P, Aib, or V. Since multiple positions are utilized to decrease hSCTR potency, even if not optimal from a selectivity perspective, X 10The position can also contain amino acids that prioritize high hGLP1 R potency. Thus, by Figure 5A It is seen that X 10 position can be selected as V, I, Y or W to maintain maximum hGLP1 R potency with varying degrees of reduction in hSCTR potency (see Figure 5A position is selected as P, Aib, V, I, Y or W to reduce hSCTR potency of the SCT scaffold. In a more preferred embodiment, X 10 position is selected as V, I, Y or W to reduce hSCTR potency of the SCT scaffold. In an even more preferred embodiment, X 10 position is selected as V, I, Y or W to reduce hSCTR potency of the SCT scaffold. In an even more preferred embodiment, X 10 position is selected as V, I or Y to reduce hSCTR potency of the SCT scaffold. In a most preferred embodiment, X 10 position is selected as V to reduce hSCTR potency of the SCT scaffold.

[0066] X 14 position: X 14 position was found to be critical for reducing hSCTR potency to provide selective GLP1 R agonists (see Example 1). A deep mutational scan showed that non-GLP-1 amino acids F, Y, W, H, Aib, G, D, A, Q, S, T, V, I, E, K, N and P were found to reduce hSCTR potency compared to the amino acid R found at X 14 position of SCT (see Example 2, Figure 6A ). To achieve the highest selectivity of GLP1 R over hSCTR, at X 14 position, preferably, the reduction in hSCTR potency should be accompanied by as little sacrifice in hGLP1 R potency as possible. In Figure 6B , the amino acids are ranked by EC 50 ratio (i.e. hSCTR EC 50 / hGLP1 R EC 50 ). The higher the EC 50 ratio of a given amino acid, the higher the selectivity for GLP1 R. Thus, when maximizing selectivity using X 14 position, preferably, X 14 is selected from F, Y, W, L, H or Aib (EC 50 ratio threshold = 0.3), more preferably, X 14 is selected from F, Y, W or L (EC 50 ratio threshold = 0.4), even more preferably, X 14 is selected from F, Y or W (EC 50 ratio threshold = 0.6), most preferably, X 14selected from F or Y (EC 50 Ratio = 0.9). Since multiple sites are utilized to reduce hSCTR potency, X 14 may also contain amino acids that prioritize high hGLP1R potency. Thus, by Figure 6A it can be seen that X 14 may be selected as F, Y, W, H, or K to maintain maximum hGLP1R potency with varying degrees of reduction in hSCTR potency (see Figure 6A ). Thus, in one embodiment, X 14 is selected as F, Y, W, L, H, Aib, or K to reduce hSCTR potency of the SCT scaffold. In a preferred embodiment, X 14 is selected as F, Y, W, L, H, or K to reduce hSCTR potency of the SCT scaffold. In a more preferred embodiment, X 14 is selected as F, Y, W, L, or K to reduce hSCTR potency of the SCT scaffold. In a most preferred embodiment, X 14 is selected as L or K.

[0067] X 18 position: X 18 position was found to be critical for increasing hGLP1R potency (see Example 1). Deep mutational scanning revealed that amino acids A, Aib, and L produced the highest potency compared to other tested amino acids at X 18 position (see Example 2, Figure 7 ). Thus, in one embodiment, X 18 position is selected as N, G, S, V, T, I, A, Aib, or L. In a preferred embodiment, X 18 position is selected as S, V, T, I, A, Aib, or L. In a more preferred embodiment, X 18 position is selected as T, I, A, Aib, or L. In an even more preferred embodiment, X 18 position is selected as A, Aib, or L to increase hGLP1R potency of the SCT scaffold. In a most preferred embodiment, X 18 position is selected as Aib.

[0068] X 19 position: X 19 position was found to be critical for reducing hSCTR potency to provide a selective GLP1R agonist (see Example 1). Deep mutational scanning revealed that, in addition to amino acid A of GLP-1 (Example 1), X 19compared to the amino acid L, all non-GLP-1 amino acids I, V, W, F, Y, R, T, K, Q, H, Aib, G, E, S, N, P and D were found to reduce hSCTR potency (see Example 2, Figure 8A ). To achieve the highest selectivity of GLP1R over hSCTR, at position X 19 , preferably, the reduction of hSCTR potency should be accompanied by as little sacrifice of hGLP1R potency as possible. In Figure 8B , the amino acids are ranked by selectivity (i.e. hSCTR EC 50 / hGLP1R EC 50 ). The higher the EC 50 ratio of a given amino acid, the higher the selectivity for GLP1R. Thus, when maximizing selectivity using position X 19 , preferably, X 19 is selected from N, G, S, T, Aib, A, E, Q, D or P (EC 50 ratio threshold = 0.2), more preferably, X 19 is selected from N, G, S, T, Aib, A, E or Q (EC 50 ratio threshold = 0.3), even more preferably, X 19 is selected from N, G, S, T, Aib, A or E (EC 50 ratio threshold = 0.4), still more preferably, X 19 is selected from N, G, S, T, Aib or A (EC 50 ratio threshold = 0.5), most preferably, X 19 is selected from N (EC 50 ratio = 0.6). Since multiple positions are utilized to reduce hSCTR potency, position X 19 may contain amino acids that prioritize high hGLP1R potency even if not optimal from a selectivity perspective. Thus, as Figure 8A can be seen, position X 19 may be selected from I, V and T to maintain maximum hGLP1R potency while hSCTR potency is reduced to varying degrees (see boxes in Figure 8A ). Thus, in a more preferred embodiment, position X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V. The inventors found that peptides perform better in in vivo models when position X 19 is selected as L. Thus, in a most preferred embodiment, position X 19 is selected as L.

[0069] Position X 22 : X 22The position was found to be crucial for hGLP1R efficacy (see Example 1). Deep mutation scanning revealed that, compared with SCT, X 22 Compared to the amino acid L found at position X, X 22 The amino acids F, W, and Y at position F produce the highest hGLP1R efficiency (see [link]). Figure 9 Therefore, in a more preferred embodiment, X 22 The position is selected as F, W, or Y to improve the hGLP1R performance of the SCT skeleton. In the most preferred embodiment, X 22 The bit selection is F.

[0070] Alternative positions in the SCT scaffold for providing high potency and selective GLP1R agonists

[0071] X 12 Position: Deep mutation scan shows, X 12 The site is a further site for enhancing the GLP1R selectivity of the peptide. As seen in Example 2, the X site of SCT... 12 Compared to the amino acids R found at the 1st site, amino acids Y, F, W, E, G, V, Aib, I, A, L, T, H, P, D, N, S, Q, and K can reduce hSCTR efficiency, while hGLP1R efficiency is only slightly reduced or even slightly improved (see [link to relevant documentation]). Figure 10A ). Figure 10B According to EC 50 The ratio (i.e., hSCTR EC) 50 / hGLP1R EC 50 Sort the amino acids. The EC50 of a given amino acid is... 50 The higher the ratio, the higher the selectivity. Therefore, to reduce the hSCTR efficiency of the SCT skeleton, preferably, X 12 Selected from Y, F, W, E, G, V, Aib, I, A, L, T, H, P, D or N (EC) 50 (Ratio threshold = 0.2), more preferably, X 12 Selected from Y, F, W, E, G, V or Aib (EC 50 (Ratio threshold = 0.3), or even more preferably, X 12 Selected from Y, F, W or E (EC) 50 (Ratio threshold = 0.4), and more preferably, X 12 Selected from Y, F or W (EC) 50 (Ratio threshold = 0.5), or even more preferably, X 12 Selected from Y or F (EC) 50 (Ratio = 0.6). In the most preferred embodiment, X 12 Selected from Y. Table 4 shows the effects of introducing Y, F, W, E, G, V or Aib compared to the closest prior art peptide GUB06-046.

[0072] X 25 Position: Deep mutation scan shows, X 25 The site is a further site for enhancing the GLP1R selectivity of the peptide. As seen in Example 2, the X site of SCT... 25 Compared to amino acid G, amino acids S, Q, T, N, Aib, E, V, K, R, A, I, L, H, F, Y, and W were found to improve hGLP1R efficiency, while also having varying degrees of influence on hSCTR efficiency (see [link to relevant documentation]). Figure 11A ). Figure 11B According to EC 50 The ratio (i.e., hSCTR EC) 50 / hGLP1R EC 50 Sort the amino acids. The EC50 of a given amino acid is... 50 The higher the ratio, the higher the selectivity. Therefore, preferably, X 25 Selected from W, F, Y, H, A, I, K, R or Aib (EC) 50 Ratio threshold = 0.1). Most preferably, X 25 Selected from W, F, Y, H or K (EC) 50 Ratio threshold = 0.2). In the most preferred embodiment, X 25 Choose H or K.

[0073] Positions that introduce lipids without compromising hGLP1R potency

[0074] The optimal lipoylation sites were determined by introducing lipoylated lysine residues at various sites in native GLP-1 (see Example 3). Figure 12 It can be seen that X 14 Position and X 25 The X site was found to be the optimal lipoylation site without impairing hGLP1R efficacy. Therefore, in the most preferred embodiment of the invention, in order to lipoylate the peptide to improve PK (pharmacokinetic) properties, X 14 bit or X 25 The bit selection is K.

[0075] Positions that improve solubility without compromising hGLP1R potency

[0076] To improve peptide solubility, sites tolerant to glutamate (E) residues were identified (see Example 4). Figure 13A As shown, X 12 X 16 X 17 X 20 X 21 X 24 X 25 and X 27positions were found to be suitable sites for introducing glutamic acid to improve peptide solubility with minimal adverse effect on GLP1 R potency. Thus, in one embodiment of the application, one or more of positions X 12 , X 16 , X 17 , X 20 , X 21 , X 24 , X 25 and X 27 are selected as E. In a preferred embodiment, one or more of positions X 12 , X 16 , X 20 or X 24 are selected as E. In particular, positions X 16 and X 24 were found to be the optimal sites for introducing glutamic acid residues with a slight positive effect on GLP1 R potency (see Figure 13A ). Thus, in a highly preferred embodiment, positions X 16 and / or X 24 are selected as E. In a most preferred embodiment, positions X 16 and X 24 are selected as E.

[0077] Positions that improve physical stability without compromising hGLP1R potency

[0078] As can be seen from Figure 14A , Aib in position X 18 and / or X 19 reduces the fibril forming tendency of the peptide at pH 7. Likewise, E in position X 21 reduces the fibril forming tendency of the peptide at pH 7. As can be seen from Figure 14B , Aib in position X 18 and / or X 19 has no adverse effect on GLP1 R potency, whereas E in position X 21 has a slight adverse effect on GLP1 R potency. Thus, in a preferred embodiment, one or more of positions X 18 , X 19 and / or X 21 are utilized to reduce the fibril forming tendency of the peptide by introducing Aib in position X 18 , Aib in position X 19 and / or E in position X 21 . In a preferred embodiment, one of positions X 18 , X 19 or X 21 is utilized to reduce the fibril forming tendency of the peptide by introducing Aib in position X 18 , Aib in position X 19position X is Aib or at X 21 position X is E to reduce the fibril forming tendency of the peptide. In a most preferred embodiment, X 18 position X is selected to be Aib to reduce the fibril forming tendency of the peptide.

[0079] In a first aspect, the present application relates to a polypeptide comprising 27 amino acid residues (X 1 -X 27 ) having at least 50% homology to human Secretin (SCT), or a pharmaceutically acceptable salt thereof, wherein

[0080] X 2 is selected to be A, Aib or G;

[0081] X 3 is selected to be I, H, L, W, V, Y, E or Q;

[0082] X 9 is D;

[0083] X 14 is selected to be F, Y, W, L, H, Aib or K;

[0084] X 18 is selected to be A, Aib or L;

[0085] X 19 is selected to be L, N, G, S, T, Aib, A, E, Q, I or V;

[0086] X 22 is selected to be F, W or Y;

[0087] characterized in that X 10 is selected to be P, Aib, V, I, Y or W and the SCTR EC 50 / GLP1R EC 50 ratio of said polypeptide is at least 100.

[0088] Preferred embodiments of the first aspect

[0089] In a preferred embodiment, X 2 is selected to be A, Aib or G; X 3 is selected to be I, H, L, E or Q; X 9 is selected to be D; X 10 is selected to be V, I, Y or W; X 14 is selected to be F, Y, W, L or K; X 18 is selected to be A, Aib or L; X 19 is selected to be L, N, G, S, T, Aib, A, E, Q, I or V; X 22is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib, or G; X 3 is selected from I, H, L, E, or Q; X 9 is selected from D; X 10 is selected from V, I, Y, or W; X 14 is selected from F, Y, W, L, or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F, W, or Y. In another preferred embodiment, X 2 is selected from A, Aib, or G; X 3 is selected from I, H, L, E, or Q; X 9 is selected from D; X 10 is selected from V, I, Y, or W; X 14 is selected from F, Y, W, L, or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib, or G; X 3 is selected from I, H, L, E, or Q; X 9 is selected from D; X 10 is selected from V, I, Y, or W; X 14 is selected from L or K; X 18 is selected from A, Aib, or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 is selected from F, W, or Y. In another preferred embodiment, X 2 is selected from A, Aib, or G; X 3 is selected from I, H, L, E, or Q; X 9 is selected from D; X 10 is selected from V, I, Y, or W; X 14 is selected from L or K; X 18 is selected from A, Aib, or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib, or G; X 3 is selected from I, H, L, E, or Q; X 9 is selected from D; X 10is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from A, Aib or G; X 3 is selected from E; X 9 is selected from D; X 10 is selected from V; X 14 is selected from L or K; X 18 is selected from Aib; X19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X14 is selected from L or K; X 18 is selected from A, Aib or L; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected from F. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V, I, Y or W; X 14 is selected from L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F, W or Y. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X 18 is selected from Aib; X 19 is selected from L; X 22 is selected from F. In another preferred embodiment, X 2 is selected from Aib; X 3 is selected from I, H, L, E or Q; X 9 is selected from D; X 10 is selected from V; X 14 is selected from F, Y, W, L or K; X18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Choose V, I, Y, or W; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V; X 22 Choose F. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X19 Choose L; X 22 Choose F, W, or Y. In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F.

[0090] In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 The choice is F, W, or Y, or in a highly preferred embodiment, X. 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22F is selected. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14Choose F, Y, W, L, or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose F, Y, W, L, or K; X 18 Select Aib; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 F is selected. In a highly preferred embodiment, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 The choice is F, W, or Y. In a highly preferred embodiment, X 2 Select Aib; X3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F.

[0091] In the most preferred embodiment of the first aspect, X 2 Select Aib; X 3 Choose E; X 9 Choose D; X 10 Select V; X 14 Choose L or K; X 18 Select Aib; X 19 Choose L; X 22 Choose F.

[0092] In any of the above embodiments of the first aspect, X 12 The optimal choice is Y, X 25 The most preferred choice is K or H.

[0093] In any implementation, X 14 When K is chosen, this lysine residue serves as a lipidation site during peptide lipoylation. Similarly, in any embodiment, X... 25 Choosing K allows the lysine residue to serve as a lipidation site during peptide lipoylation. Most preferably, only X... 14 or X 25 One option is K, such that the peptide contains only a single lipid-modified lysine residue. Therefore, in any of the above embodiments, when X... 14 When K is chosen to provide lipidation sites, X is the most preferred. 25 Choose H. Similarly, in any of the above embodiments, when X... 25 When K is chosen to provide lipidation sites, X is the most preferred. 14 Choose L.

[0094] In a second aspect, the present invention relates to a polypeptide or a pharmaceutically acceptable salt thereof, comprising the structure of formula (I),

[0095]

[0096] in,

[0097] X 2 Choose A, Aib, or G;

[0098] X 3Choose from I, H, L, W, V, Y, E, or Q;

[0099] X 12 Choose from Y, F, W, E, G, V, Aib, I, A, L, T, H, P, D, N, or R;

[0100] X 14 Choose F, Y, W, L, H, Aib, or K.

[0101] X 16 Choose E, G, T, F, L, or Aib;

[0102] X 18 Choose A, Aib, or L;

[0103] X 19 Choose from L, N, G, S, T, Aib, A, E, Q, I, or V;

[0104] X 22 Choose F, W, or Y;

[0105] X 23 Choose either L or I;

[0106] X 24 Choose either E or Q;

[0107] X 25 Choose W, F, Y, H, K, or E;

[0108] Its characteristic is that X 10 Choose from P, Aib, V, I, Y, or W.

[0109] Embodiments of the second aspect

[0110] It should be understood that X 2 X 3 X 10 X 12 X 14 X 16 X 18 X 19 X 22 X 23 X 24 and X 25Each amino acid site in the above headings is selected from one of the amino acids or groups of amino acids described under the headings “Key SCT backbone sites for providing highly potent and selective GLP1R agonists,” “Optional SCT backbone sites for providing highly potent and selective GLP1R agonists,” “Sites for introducing lipids without impairing hGLP1R efficacy,” and “Sites for improving solubility without impairing hGLP1R efficacy.” Therefore, any combination of various embodiments under these headings is covered, whether that embodiment is “preferred,” “more preferred,” or “most preferred.”

[0111] Preferred embodiments of the second aspect

[0112] The first aspect lists the specified X 2 X 3 X 10 X 14 X 18 X 19 and X 22 The preferred amino acid embodiment is also applicable to the second aspect. Therefore, any embodiment mentioned under the heading "Preferred Embodiment of the First Aspect" is also applicable to the second aspect. Furthermore, for each mentioned embodiment, it further includes: preferably, X 12 Selected from Y, F, W, E, G, V, or Aib, or even more preferably, X 12 Selected from Y, F, W, or E, and more preferably X 12 Selected from Y, F, or W, with X being the most preferred. 12 Choose Y; X 16 Choose from E, G, T, F, L, or Aib, with E being the optimal choice; X 23 Choose either L or I, with L being the optimal choice; X 24 Choose either E or Q, with E being the preferred option.

[0113] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0114] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0115] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0116] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0117] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0118] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0119] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0120] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F; X 23 Choose L; X24 Choose E; X 25 Choose H or K.

[0121] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0122] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0123] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0124] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0125] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0126] In another preferred embodiment, X 2 Choose A, Aib, or G; X 3 Choose E; X 10 Select V; X12 selects Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0127] In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y; X23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0128] In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0129] In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0130] In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0131] In another preferred embodiment, X 2Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0132] In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0133] In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0134] In another preferred embodiment, X 2 Select Aib; X 3 Choose I, H, L, E, or Q; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0135] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0136] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0137] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0138] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Choose V, I, Y, or W; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0139] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0140] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Choose A, Aib, or L; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0141] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X18 Select Aib; X 19 Choose L; X 22 Choose F, W, or Y; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0142] In another preferred embodiment, X 2 Select Aib; X 3 Choose E; X 10 Select V; X 12 Choose Y, F, or V; X 14 Choose L or K; X 16 Choose E; X 18 Select Aib; X 19 Choose L; X 22 Choose F; X 23 Choose L; X 24 Choose E; X 25 Choose H or K.

[0143] As shown in Tables 1 and 2 of this article, the SCTR EC of exemplary peptides 50 / GLP1R EC 50 The ratio is at least 100, and the ratio of most peptides is much higher than 100. As can be seen from Table 3, compared to the prior art peptide GUB06-046, X... 10 The presence of amino acids P, Aib, V, I, Y, or W at position X can improve selectivity. Similarly, as shown in Table 4, compared to the prior art peptide GUB06-046, X... 12 The presence of amino acids Y, F, W, E, G, V, or Aib at position Y can improve selectivity. Therefore, in the most preferred embodiment, the polypeptide is characterized by its SCTREC... 50 / GLP1R EC 50 The ratio must be at least 100.

[0144] In any implementation of the second aspect, if X 14 If K is selected, then the lysine residue serves as a lipidation site when the peptide is lipidated. Similarly, in any embodiment of the second aspect, if X... 25 Choosing K means that the lysine residue serves as a lipidation site when the peptide is lipidated. Most preferably, only X... 14 or X 25 One option is K, such that the peptide contains only a single lipid-modified lysine residue. Therefore, in any embodiment of the second aspect, when X... 14 When K is chosen to provide lipidation sites, X is the most preferred. 25Choose H. Similarly, in any of the above embodiments, when X... 25 When K is chosen to provide lipidation sites, X is the most preferred. 14 Choose L. The most preferred option is X. 14 K was chosen as the site for lipidation.

[0145] Tables 1 and 2 illustrate the selective GLP1R agonists of the present invention, whose X... 2 X 3 X 9 X 10 X 14 X 18 X 19 and X 22 Amino acid residues at the identified sites can convert SCT into a selective GLP1R agonist. The last row of Table 1 summarizes exemplary amino acids at the identified sites.

[0146] Extended GLP1R agonists based on SCT

[0147] As shown in Example 7 and Table 2, the GLP1R agonist of the present invention can be extended to the full length of natural GLP-1 (i.e., 30 amino acid residues). Therefore, in any aspect of the first, second, or third aspect or any embodiment thereof, the peptide can be extended by 1 (i.e., X). 28 ), 2 (i.e., X) 28 -X 29 ) or 3 (i.e., X) 28 -X 29 -X 30 Additional amino acid residues. When extended, X 28 The amino acid at position X can be selected from any one of Aib, A, D, E, F, G, H, I, K, L, N, P, Q, R, S, T, V, W, or Y. However, X 28 Choosing F or P is less preferred. Preferably, X 28 Choose either A or E. The most preferred option is X. 28 Choose E. X 29 The amino acid at position X can be selected from any one of Aib, A, D, E, F, G, H, I, K, L, N, P, Q, R, S, T, V, W, or Y. However, most preferably, X 29 Choose A, Aib, or G. X 30 The amino acid at position X can be selected from any one of Aib, A, D, E, F, G, H, I, K, L, N, P, Q, R, S, T, V, W, or Y. However, most preferably, X 30 Choose either A or E. Therefore, in a highly preferred embodiment, when the GLP1R agonist of the present invention comprises 30 amino acid residues, X28 Choose A or E; X 29 Choose A, Aib, or G; and X 30 Choose either A or E. In the most preferred embodiment, when the GLP1R agonist of the present invention contains 30 amino acid residues, X 28 Choose E; X 29 Choose A; and X 30 Choose A.

[0148] Medical uses

[0149] In a fourth aspect, the present invention relates to the use of the peptides in any of the foregoing aspects or embodiments as a medicament, particularly as a medicament for treating obesity, diabetes and / or kidney disease and / or non-alcoholic steatohepatitis (NASH), most preferably as a medicament for treating obesity and / or diabetes.

[0150] Experimental Section

[0151] Conventional methods of peptide synthesis

[0152] Peptide synthesis was performed using a SyroII fully automated parallel peptide synthesizer (MultiSynTech GmbH, Germany) equipped with a heating block, on Tentagel S RAM resin (Rapp polymer GmbH, Germany) with a loading of 0.23–0.25 mmol / g. α -Fmoc deprotection was performed in two steps: the resin was treated with 40% piperidine / DMF (0.2 M HOBt (1-hydroxybenzotriazole)) at 45°C for 3 minutes, followed by treatment with 20% piperidine / DMF (0.1 M HOBt) at 75°C for 7 minutes. Except for Asp, Cys, and His residues, the N... α -Fmoc deprotection was performed at room temperature, i.e., treatment with 40% piperidine / DMF (0.2 M HOBt) for 3 minutes, followed by treatment with 20% piperidine / DMF (0.1 M HOBt) for 15 minutes. Coupling chemistry used a DMF solution of DIC (N,N'-diisopropylcarbodiimide) / Oxyma (ethyl cyano(hydroxyimino)acetate), with a 0.5 M DMF solution of the amino acid and a 5-fold molar excess. Standard Fmoc-protected amino acids were used. Coupling conditions were single or double coupling at 75 °C for 15 minutes. His and Cys residues were excluded, and these residues were double-coupled at 50 °C for 15 minutes. Additionally, amino acids coupled after Aib were also double-coupled. Fmoc amino acids were dissolved in 0.5 M DMF solution containing 0.5 M Oxyma, except for His, which was dissolved in NMP.α -Fmoc deprotected the resin and washed it 5 times with DMF, then washed it 3 times after coupling.

[0153] For N-terminal lipolation examples, lipolation is performed on a resin as the final step in peptide synthesis. N-terminal lipolation peptides optionally include linker residues such as [γE], [OEG], [OEG]-[OEG], etc. Linker residues are introduced by coupling Fmoc-OEG-OH, Fmoc-OEG-OEG, and / or Fmoc-Glu-OtBu to the N-terminal amino acid before coupling with a fatty diacid (e.g., tert-butyl-protected fatty diacids such as tBu-C18-diacid, tBu-C20-diacid), thereby elongating the peptide chain. Linker residues are double- or triple-coupled under standard conditions. Fatty acids are double-coupled using 2 equivalents of building blocks.

[0154] For other lipoylation examples, the Boc-protected amino acid was introduced as an N-terminal residue, and the lipoylation site was introduced as an orthogonally protected lysine (Lys(Mtt)). The Mtt group (4-methyltriphenylmethyl) was removed by treating the resin with a DCM (dichloromethane) solution of 75% HFIP (1,1,1,3,3,3-hexafluoroprop-2-ol) supplemented with 5% TIPS for 10 minutes; this step was repeated three times. The resin was washed with a DCM solution of 10% DIPEA, followed by three washes with DMF.

[0155] The lipotropic peptide optionally includes linker residues. The linker residues are coupled to the ε-amino group of a deprotected lysine residue before coupling to a fatty acid (e.g., tert-butyl-protected fatty diacids such as tBu-C18-diacid or tBu-C20-diacid). The linker residues are double- or triple-coupled under standard conditions. Double coupling of the fatty acid is performed using two equivalents of the building block.

[0156] After synthesis, the resin was washed with DCM and dried. The peptide was excised from the resin by treatment with TFA (trifluoroacetic acid) / TES (triethylsilane) / DODT / water (93 / 2.5 / 2.5 / 2.0) at 40 °C for 45 min, followed by precipitation with 3 volumes of cold diethyl ether, washing with ether, and allowing to dry. The peptide was characterized by LC-MS (Waters, Denmark) and quantified by LC-CAD (Thermo Fisher Scientific, Denmark). Finally, the peptide was freeze-dried using a Telstar benchtop freeze dryer to obtain a white powder. The crude library peptide was used for hGLP1R and hSCTR efficacy screening.

[0157] Conventional purification of peptides

[0158] The crude peptides were dissolved in acetonitrile / water and purified by reversed-phase HPLC using a Waters preparative HPLC system equipped with a C18 column (Reprosil Gold 200 Å, 5 μm, 30 mm × 250 mm), a preparative pump (Waters 2545), a UV / Vis detector (Waters 2489), and a Waters fraction collector III. The mobile phase consisted of a gradient elution of buffer A (0.1% TFA aqueous solution) and buffer B (0.1% TFA acetonitrile solution) at a flow rate of 20 mL / min at room temperature. The fractions were analyzed, combined, and lyophilized. Finally, the peptides were lyophilized using a Telstar benchtop freeze dryer. The purity and molecular weight of peptides were determined by analytical RP-HPLC-MS on an ACQUITY UPLC Peptide CSH C18 column (Waters, ACQUITY UPLC Peptide CSH, C18, 130 Å, 1.7 μm, 2.1 mm × 100 mm) using a Waters Acquity HPLC system equipped with a 3100 mass detector. Analysis was performed at 40 °C using gradient elution with buffer A (0.3% TFA aqueous solution) and buffer B (0.3% TFA acetonitrile solution) (gradient of 40-60% B over 14 minutes).

[0159] Conventional handling of peptides for in vitro potency testing

[0160] The crude library peptides were screened using 5 10-fold serial dilutions at different concentrations, n=1. Pipe tips were changed for the first two serial dilutions. If necessary, the initial concentration (maximum concentration) of the compound was adjusted throughout the study.

[0161] The purified peptides were screened using 11 3-fold serial dilutions, n=2. Pipe tips were changed for each serial dilution. The initial concentration (maximum concentration) of the compound was adjusted throughout the study. If the maximum concentration was adjusted, or if the calculated EC was observed... 50 If the values ​​differ by a factor of 5, then increase n by 1. EC 50 The values ​​were calculated using a nonlinear regression with a variable-slope sigmoid concentration-response curve. As shown in Table 6, the efficacy data obtained from the purified peptide showed a good correlation with those obtained from the crude peptide.

[0162] Conventional procedures for determining hGLP1R potency

[0163] CHO-K1 cell lines stably overexpressing hGLP1R were obtained from Euroscreen (FAST-0145L), expanded, aliquoted, and frozen. One unit of cells was thawed and seeded at 2000 cells per well in DPBS containing 0.05% casein and 0.5 mM IBMX into 384-well plates. Cells were then immediately stimulated for 30 minutes at room temperature with gradient doses of the test compound. Human GLP-1 (7-36) (synthesized by Gubra) was used as a positive control. cAMP accumulation was measured using the Cisbio Gs-coupled receptor assay kit (product number 62AM4PEC), where the assay reagent was added according to the manufacturer's instructions, and time-resolved fluorescence resonance energy transfer was recorded on a CLARIOstar (BMGLabtech) plate reader 1 hour later.

[0164] Conventional procedures for determining hSCTR potency

[0165] CHO-K1 cells stably overexpressing hSCTR and frozen with arrested cell division were obtained from PerkinElmer (ES-712-AF). Alternatively, CHO-K1 cell lines stably overexpressing hSCTR were obtained from Euroscreen (FAST-0161L), expanded, aliquoted, and frozen. One unit of cells was thawed and seeded at 2000 cells per well in DPBS containing 0.05% casein and 0.5 mM IBMX into 384-well plates. Cells were then immediately stimulated for 30 minutes at room temperature with gradient doses of the test compound, with human secretin (synthesized by Gubra) used as a positive control. cAMP accumulation was measured using the Cisbio assay kit for Gs-coupled receptors (product number 62AM4PEC), where the assay reagent was added according to the manufacturer's instructions, and time-resolved fluorescence resonance energy transfer was recorded on a CLARIOstar (BMG Labtech) plate reader 1 hour later.

[0166] Conventional procedures for determining turbidity and fibril formation of crude library peptides

[0167] The peptide was dissolved in buffer (50 mM sodium acetate buffer, pH 4.0 or 50 mM sodium phosphate buffer, pH 7.0) at a concentration of 267 μM and incubated at room temperature for 1–2 hours. The sample was then aliquoted into two 80 μL replicates and dispensed into black 384-well plates (μ-clear, Greiner Bio-One), mixed with thioflavin T (ThT) to a final concentration of 4 μM. The plates were centrifuged at 2000 rpm for 2 minutes to remove air bubbles, sealed, and placed in a plate reader (CLARIOstar, BMG). First, the turbidity of the sample was measured at 600 nm as absorbance. Second, the plate reader temperature was set to 40 °C, and fluorescence was measured every 10 minutes by exciting ThT at 450 nm and measuring the emission intensity at 480 nm for 72 hours. Before each measurement, the sample was stressed by shaking the plate at 700 rpm (linearly) for 5 minutes, and fibril formation for each sample was expressed as average emission intensity.

[0168] Conventional procedures for determining fibril formation of purified peptides

[0169] Detection of thioflavin T fibril formation: The peptide was dissolved in 50 mM phosphate buffer (pH 7.0) at room temperature for 2 hours on a shaker at room temperature to a concentration of 267 μM. The sample was then aliquoted into three replicates of 80 μL / well and mixed with 2 μL / well of thioflavin T (ThT). The mixture was then dispensed into black 384-well microplates (Greiner, product number 781096) with a clear bottom, resulting in a final concentration of 4 μM. The plates were inserted into a CLARIOstar Plus microplate reader (BMGLabtech), and the cycling conditions were 5 minutes of incubation followed by 5 minutes of linear shaking at 700 rpm, followed by 96 hours at 40°C. Fibril formation was measured as an increase in fluorescence emission intensity at 480 nm (excited at 450 nm).

[0170] Conventional procedures for determining solubility of purified peptides

[0171] Solubility was tested in the following carrier: 100 mM phosphate buffer, with a target pH of 7.0. The sample was dissolved directly in a 0.45 μm unit Mini-UniPrep needleless filter (Whatman). For the desired concentration (e.g., approximately 10 mg / mL), 1050 nmol of peptide was dissolved in 400 μL of carrier. The sample was incubated on a shaker at room temperature for at least 1 hour. The pH was measured and adjusted. The sample was incubated on a shaker for another hour, and the pH was measured and adjusted again. A visual inspection was performed and recorded before pushing the filter. The concentration of peptide in the filtrate was determined using a CAD (electro-fogging detector), with two measurements performed for each sample. The measured concentrations are expressed in μM. Additionally, the pH of the filtrate was measured and recorded. Peptides with a measured concentration within ±20% of the target concentration were considered completely soluble.

[0172] Example 1 : Identification of key amino acid residues in SCT that improve hGLP1R potency and reduction hSCTR potency

[0173] Based on the natural sequence of SCT, a peptide library was designed and synthesized, in which, when compared as shown below, each amino acid residue in SCT that differs from GLP-1 (i.e., X) 2 -X 3 X 9 -X 10 X 12 -X 14 and X 17 -X 25 The position is changed to the corresponding GLP-1 residue (one or a combination of substitutions at a time).

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[0175] This provided a library of 735 peptides, including 692 peptides with 1–6 GLP-1 substitutions in the SCT backbone and 43 peptides with 14–16 GLP-1 substitutions in the SCT backbone. ECG measurements of hGLP1R and hSCTR were performed. 50 The value is calculated using a random forest model, where the EC value is used to calculate the SHAP value. 50The SHAP value was fitted to the amino acid sequence of the peptide. The SHAP value was used to determine the contribution of each amino acid substitution to the two endpoints of hGLP1R and hSCTR efficacy (Breiman, L. (2001), Random Forests, Machine Learning 45(1), 5-32; Lundberg, SM, and Lee, SI (2017); A unified approach to interpreting model predictions, Advances in neural information processing systems, 30). The contribution of each substitution was determined by the difference in average SHAP values ​​between SCT amino acids and GLP-1 amino acids. Substitutions with positive SHAP values ​​improved the endpoints, while substitutions with negative SHAP values ​​reduced the endpoints.

[0176] Figure 1A and Figure 1B The SHAP values ​​of different amino acid sites relative to hGLP1R and hSCTR efficacy are shown. Figure 1A It can be seen that X 2 X 9 X 18 and X 22 These sites were found to be the most important for improving hGLP1R performance because they had the highest positive SHAP values ​​(threshold set at 0.1). Similarly, by Figure 1B It can be seen that X 3 X 9 X 10 X 14 and X 19 The sites were found to be most important for eliminating hSCTR effectiveness (i.e., providing selectivity) because these sites had the lowest negative SHAP values ​​(threshold set at 0.1).

[0177] Example 2: Identification of non-GLP-1 residues in SCT that are able to improve hGLP1R potency and reduction hSCTR potency

[0178] A peptide library of GLP1R agonists based on the SCT backbone was designed and synthesized, wherein each amino acid residue of SCT (i.e., X) is present. 1 -X 27All positions were altered to all other natural amino acids or Aibs except for the corresponding sites present in SCT and GLP-1 (one or a combination of substitutions at a time), referred to herein as deep mutation scans. This provided a library of 854 peptides, each containing an SCT backbone with 1–4 substitutions. The inventors constructed a random forest model (Breiman, L. (2001), Random Forests, Machine Learning 45(1), 5–32) to describe the relationship between peptide sequences and the efficacy of hGLP1R and hSCTR. In addition to the sites identified using GLP-1 amino acids in Example 1 (i.e., X sites used to improve hGLP1R efficacy), 2 X 9 X 18 The bit, and the X used to reduce hSCTR performance to provide selectivity 22 X 3 X 9 X 10 X 14 and X 19 In addition to the (position), Example 2 also identified X 12 and X 25 This can further improve hGLP1R efficiency and reduce hSCTR efficiency. Using a random forest model, based on a large number of peptides in a peptide library, the normalization effect of altered amino acids was calculated. Data from each identified site are summarized in [the table / incomplete]. Figure 2 -Figure 8.

[0179] Example 3: Identification of lipidation sites that do not compromise hGLP1R potency

[0180] The optimal lipidation sites were identified by introducing lipidated lysine residues at various sites in native GLP-1. Figure 12 As shown, lipidation of lysine residues at positions 14 and 25 has the least adverse effect on hGLP1R efficacy. Therefore, in a preferred embodiment of the invention, X 14 or X 25 Lysine (K) was chosen, wherein lysine was lipidized to alter the PK properties of the peptide.

[0181] Example 4: Identification of sites for increasing solubility without compromising hGLP1R potency

[0182] To improve peptide solubility, sites tolerant to glutamate (E) residues (i.e., those not impairing hGLP1R efficacy) were identified. Using the amino acids identified in Examples 1 and 2, a library of GLP1R agonists based on the SCT backbone was constructed. Furthermore, the peptides in this library were found to be resistant to X-ray diffusivity. 9 -X 27 The peptides in this library have 1-2 glutamic acid (E) substitutions. hGLP1R efficacy was determined. Peptides in this library were lipid-modified at positions 10, 14, 17, or 25.Figure 13A It can be seen that X 12 X 16 X 17 X 20 X 21 X 24 X 25 and X 27 The optimal position was identified, as it had a minimal adverse effect on hGLP1R efficacy, and even a slight improving effect. Specifically, X... 16 and X 24 The position was identified as the optimal site for introducing glutamate residues, with X being the most preferred. 16 Bit.

[0183] Example 5: Identification of sites that improve physical stability

[0184] Mutations that improve the physical stability of peptides (i.e., inhibit protofibrosis) without impairing hGLP1R efficacy were identified. A library of GLP1R agonists based on the SCT backbone was constructed using the amino acids identified in Examples 1 and 2. Furthermore, the peptides in this library exhibited X... 3 X 10 X 12 X 18 X 19 or X 21 The peptides in this library have 1-3 mutations at position X. 14 The site is lipid-modified. Data is summarized in Figure 14A and Figure 14B .

[0185] Example 6: Exemplary GLP1R agonists of the invention having 27 amino acid residues

[0186] Based on the key findings of Examples 1-5, the library was designed, synthesized, and tested. Table 1 shows the synthesized and tested peptides. It can be seen that the peptides incorporating the amino acids from Examples 1 and 2 are highly potent and selective GLP1R agonists.

[0187] Example 7: Exemplary extended GLP1R agonists having up to 30 amino acid residues

[0188] Since natural GLP-1 is 30 amino acids long and natural SCT is 27 amino acids long, GLP1R agonists based on the SCT backbone can be extended to the full length of natural GLP-1. To illustrate this, libraries of GLP1R agonists with lengths of 28-30 amino acids were constructed based on the SCT backbone and the mutations identified in Examples 1 and 2. These peptides were 27, 28, 29, or 30 amino acids long. X was tested. 28 X 29 and X 30The peptides in this library are Aib, A, D, E, F, G, H, I, K, L, N, P, Q, R, S, T, V, W, or Y. The peptides in this library have not been lipid-modified. hGLP1R efficacy was determined. Figure 15A As shown, besides F and P having an adverse effect on hGLP1R performance, X 28 The amino acid sequence determined at position X has almost no effect on hGLP1R efficacy. Furthermore, Figure 15 shows that X... 29 Amino acids A, Aib, and G at position A have a slight positive effect on hGLP1R efficacy, while amino acids D, E, L, and W have a slight negative effect. The remaining amino acids tested have almost no effect on hGLP1R efficacy. Finally, Figure 15A As shown, X 30 All amino acids tested were tolerated, but amino acid G had a slight positive effect on hGLP1R efficacy. Table 2 shows the GLP1R agonists of the present invention extended to the full length of natural GLP-1.

[0189] Example 8: Effect on food intake

[0190] Five-week-old male NMRI mice were purchased from JanVier (JanVier Labs, France). Animals were housed in groups of four per cage, under a 12-hour / 12-hour dark-light cycle, with lights turned off at 1 PM. Room temperature was maintained at 22℃±1℃, and humidity at 60%±20%. Animals had free access to standard rodent feed (Altromin 1324, Brogaarden, Denmark) and tap water.

[0191] Five to seven days prior to the start of the study, animals were transferred to a real-time food intake monitoring system (HM-2 system, MBRose, Denmark) to acclimatize to experimental conditions. Since animals are uniquely identified by microchips, each individual animal was identified by its microchip upon entering and leaving the food passage. Mice in each experimental group (n=7-8) were randomly assigned to groups based on their body weight measured the day before the study began. Each experiment included a loading treatment group. Animals were fasted for 6 hours prior to the start of the dark period. One hour prior to the start of the dark period, the test peptide was administered subcutaneously to the animals (see [link to study details]). Figure 16A / B and Figure 17A / B). Formulation: 20 nmol / mL peptide, dissolved in 50 mM phosphate buffer (pH 7.4) containing 3.5% mannitol. Hourly food intake was recorded over 72 hours. The percentage reduction in food intake was normalized relative to the mean food intake of the load cell group. Statistical significance was assessed using one-way ANOVA and Tukey's multiple comparison test. P < 0.05 was considered statistically significant.

[0192] Reference List

[0193] Witteloostuijn et al., J. Pept. Sci. December 2017, 23(12), pp. 845-854.

[0194] Breiman, L. (2001), Random Forests, Machine Learning 45(1), pp. 5-32.

[0195] Lundberg, SM, and Lee, SI, NIPS 2017, A Unified Approach to Interpreting Model Predictions, pp. 1-10.

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Claims

1. A polypeptide or a pharmaceutically acceptable salt thereof comprising the structure of Formula (I), wherein, 10. The polypeptide or a pharmaceutically acceptable salt thereof of claim 1, comprising the structure of Formula (I), X 2 is selected as A, Aib or G; X 3 is selected as I, H, L, W, V, Y, E or Q; X 12 selected from Y, F, W, E, G, V, Aib, I, A, L, T, H, P, D, N or R; X 14 is selected F, Y, W, L, H, Aib or K X 16 is selected as E, G, T, F, L or Aib; X 18 is selected as A, Aib or L; X 19 is selected as L, N, G, S, T, Aib, A, E, Q, I or V; X 22 is selected as F, W or Y; X 23 is selected as L or I; X 24 is selected as E or Q; X 25 is selected as W, F, Y, H, K or E; characterized in that X 10 is selected as P, Aib, V, I, Y or W.

2. The polypeptide or a pharmaceutically acceptable salt thereof of claim 1, wherein, X 3 is selected as I, H, L, E or Q.

3. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 10 is selected from V, I, Y or W.

4. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 14 X is selected from F, Y, W, L, H or K, most preferably, X 14 X is selected from L or K.

5. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 19 is selected to be L.

6. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 12 is selected from Y, F, W, E, G, V or Aib, preferably X 12 is selected from Y, F, W or E, most preferably X 12 is selected from Y.

7. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 23 Select is L.

8. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 25 X is selected from W, F, Y, H, K or E, most preferably X 25 X is selected from K or H.

9. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 12 , X 16 or X 24 is selected to be E, preferably X 16 and / or X 24 is selected to be E, most preferably X 16 and X 24 are selected to be E. the polypeptide comprises the sequence of H[Aib]EGTFTSDVSYLLEEA[Aib]LQRFLEKLVEAA (SEQ ID NO: 213); H[Aib]EGTFTSDVSYLKEEA[Aib]LQRFLEHLVEAA (SEQ ID NO: 209); or H[Aib]EGTFTSDVSYLKEEA[Aib]LQRFLEHLV (SEQ ID NO: 203). X 2 is selected as A, Aib or G; X 3 is selected as I, H, L, E or Q; X 14 selected to be L or K; X 18 is selected as A, Aib or L; X 22 is selected as F, W or Y; X 25 is selected as H or K; characterized in that X 10 is selected from V, I, Y or W.

11. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 2 is selected from Aib.

12. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 3 E is selected.

13. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 10 Select is V.

14. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 18 is selected from Aib.

15. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 22 Select is F.

16. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 14 or X 25 only one of which is selected as K, and wherein K is optionally lipidated via a linker / spacer lipid.

17. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, X 14 is selected to be K and X 25 is selected to be H, and wherein K is optionally lipidated through a linker / spacer lipid; or wherein X 14 is selected to be L and X 25 is selected to be K, and wherein K is optionally lipidated through a linker / spacer lipid.

18. The polypeptide or pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, SCTR EC of the polypeptide 50 / GLP1R EC 50 The ratio is at least 100.

19. The polypeptide or pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, The polypeptide comprises 30 amino acid residues (X 1 - X 30 ).

20. The polypeptide or a pharmaceutically acceptable salt thereof of claim 1, wherein, the polypeptide consists of the sequence of H[Aib]EGTFTSDVSYLLEEA[Aib]LQRFLE[K*]LVEAA (SEQ ID NO: 213); H[Aib]EGTFTSDVSYL[K*]EEA[Aib]LQRFLEHLVEAA (SEQ ID NO: 209); or H[Aib]EGTFTSDVSYL[K*]EEA[Aib]LQRFLEHLV (SEQ ID NO: 203), wherein * indicates covalent attachment of C20DA[γE][OEG][OEG]- to lysine.

21. The polypeptide or a pharmaceutically acceptable salt thereof of claim 1, wherein, the polypeptide is amidated (-CONH2) at the C-terminus.

22. The polypeptide or pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, the polypeptide has an amino group (-NH2) at the N-terminus.

23. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims, wherein, 24. The polypeptide or a pharmaceutically acceptable salt thereof of any one of the preceding claims for use as a medicament; preferably for use as a medicament for the treatment of obesity, diabetes, kidney disease, and / or nonalcoholic steatohepatitis (NASH); most preferably for use as a medicament for the treatment of obesity and / or diabetes.

25. A pharmaceutical composition comprising the polypeptide or a pharmaceutically acceptable salt thereof of any one of claims 1-23. ​

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