Co-agonists at GLP-1 and GIP receptors suitable for oral delivery

Abstract

To provide single molecule co-agonists comprising a peptide and a substituent, which react with both the human GLP-1 and GIP receptors with high potency and are suitable for once-daily oral administration to humans.SOLUTION: Provided are GLP-1 and GIP receptor co-agonists suitable for oral delivery achieved by the combination of certain peptide sequence variants, such as compounds with specific amino acid sequences, with substituents via a single site acylation with a diacid based fatty acid.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to compounds that are glucagon-like peptide-1 (GLP-1) receptor agonists and glucose-dependent insulinotropic polypeptide (GIP) receptor agonists with a prolonged profile of action suitable for oral administration to humans.

[0002] Incorporation by reference of sequence listing This application is submitted with a Sequence Listing in electronic format, the entire contents of which are incorporated herein by reference. [Background technology]

[0003] Glucagon-like peptide 1 (GLP-1) is an enteroendocrine cell-derived hormone and one of two prominent endogenous physiological incretins. GLP-1 improves glycemic control by stimulating glucose-dependent insulin secretion in response to nutrients (glucose) and inhibits glucagon secretion from pancreatic α-cells, slowing gastric emptying and inducing weight loss primarily by reducing food consumption. The other prominent incretin, glucose-dependent insulinotropic polypeptide (GIP), improves glycemic control by stimulating insulin secretion in response to nutrients (fat, glucose). In addition, GIP appears to improve plasma lipid profiles and stimulate calcium accumulation in bone. In contrast to GLP-1, the incretin effect of GIP is greatly reduced in type 2 diabetes patients, although recent studies suggest that GIP efficiency in these patients can be regained after treatment to improve glucose control. Nevertheless, the role of GIP in regulating whole-body metabolism beyond its direct effects in the endocrine pancreas remains controversial, especially as GIP actions in promoting fat mass gain in animal models are relevant. These results raise the belief that GIPR antagonism can improve body weight. Therefore, compounds acting at the GIP receptor, particularly agonistic or antagonistic, as a strategy to improve body weight are still the subject of intensive scientific investigation ([Non-Patent Document 1], [Non-Patent Document 2]).

[0004] In rodent models, prolonged GIP analogs have been shown to reduce body weight and improve glycemic control, but are less potent in reducing body weight compared to GLP-1 analogs ([Non-Patent Document 3]). Furthermore, GIP analogs induce weight loss by additive / synergistic action with GLP-1 analogs when administered dually ([Non-Patent Document 4], [Non-Patent Document 5]), and therefore represent suitable candidates for amplifying GLP-1-based pharmacology. As shown in preclinical animal models, GIPR agonism can be employed as a non-redundant partner of GLP-1R agonism as a single molecule coagonist to amplify GLP-1 metabolic benefits, most notably weight loss and glycemic control ([Non-Patent Document 4], [Non-Patent Document 6]). Two different peptides with potent dual incretin receptor agonism have advanced into multiple-dose clinical trials. Clinical results have shown improvements in glycemic control and body weight beyond those achieved by comparable administration of benchmark GLP-1 specific agonists ([Non-Patent Document 7], [Non-Patent Document 8]), demonstrating the translational aspects and therapeutic benefits of simultaneous targeting of the GLP-1 and GIP receptors.

[0005] Oral delivery of GLP-1 derivatives is being clinically investigated in the form of a once-daily tablet of semaglutide and the permeation enhancer sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC) to improve the very low typical exposure and bioavailability of GLP-1 derivatives following oral administration ([Non-Patent Document 9]).

[0006] GLP-1 / GIP coagonists and their potential medical uses have been described in several patent applications, such as [US Pat. No. 5,393,621], [US Pat. No. 5,493,631], [US Pat. No. 5,493,645], [US Pat. No. 6,493,706], [US Pat. No. 6,493,811], [US Pat. No. 6,493,925], [US Pat. No. 6,493,103], [US Pat. No. 6,493,114], and [US Pat. No. 6,503,312]. Patent applications disclosing oral delivery of GLP-1 derivatives are described, for example, in [US Pat. No. 6,493,625], [US Pat. No. 6,503,706], [US Pat. No. 6,493,811], and [US Pat. No. 6,503,116].

[0007] However, no co-agonist products have received marketing approval to date. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Publication No. 2010 / 011439 [Patent Document 2] International Publication No. 2013 / 164483 [Patent Document 3] International Publication No. 2014 / 192284 [Patent Document 4] International Publication No. 2015 / 067715 [Patent Document 5] International Publication No. 2015 / 022420 [Patent Document 6] International Publication No. 2015 / 086728 [Patent Document 7] International Publication No. 2015 / 086729 [Patent Document 8] International Publication No. 2016 / 111971 [Patent Document 9] International Publication No. 2020 / 023386 [Patent Document 10] U.S. Patent No. 9,745,360 [Patent Document 11] U.S. Patent Publication No. 2014 / 162945 [Patent Document 12] U.S. Patent Publication No. 2014 / 0357552 [Patent Document 13] International Publication No. 2011 / 080103 [Patent Document 14] International Publication No. 2012 / 080471 [Patent Document 15] International Publication No. 2013 / 189988 [Patent Document 16] International Publication No. 2019 / 149880 [Non-patent literature]

[0009] [Non-Patent Document 1] Finan et al,TRENDS Mol Med,2016,22(5):359-376 [Non-Patent Document 2] Killion et al,Endo Rev,2020,41(1):1-21 [Non-Patent Document 3] Mroz et al,Mol Metab,2019,20:51-62 [Non-Patent Document 4] Finan et al,Sci Transl Med,2013,5(209):209ra151 [Non-Patent Document 5] Norregaard et al, Diabetes Obes Metab,2018,20(1):60-68 [Non-Patent Document 6] Coskun et al,Mol Metab,2018,18:3-14 [Non-Patent Document 7] Frias et al,Cell Metab,2017,26(2):343-352 [Non-Patent Document 8] Frias et al, Lancet, 2018, 392(10160):2180-2193 [Non-Patent Document 9] Hedrington & Davis,Exp.Opin.Pharmacother.2019,20(2):133-141 Summary of the Invention

[0010] The present invention relates to single molecule coagonists comprising peptides and substituents that react potently with both human GLP-1 and GIP receptors and are suitable for once-daily oral administration to humans. This is achieved by combining certain peptide sequence variants with substituents via single-site acylation with diacid-based fatty acids.

[0011] One aspect of the present invention is a peptide having the following amino acid sequence: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPX 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:36) with an optional amide modification at the C-terminus, During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X39 is S or absent.

[0012] In another aspect, the invention provides a peptide having the amino acid sequence: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPX 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:36) with an optional amide modification at the C-terminus, In the formula, X2 is Aib or A; X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent, and a substituent bound to the peptide via lysine (K) at position 16 or 33; or a pharma- ceutically acceptable salt thereof.

[0013] A further aspect of the present invention relates to methods for preparing the GLP-1 / GIP coagonists described herein.

[0014] In a further aspect, the present invention relates to pharmaceutical compositions comprising the GLP-1 / GIP coagonist compounds described herein.

[0015] A further aspect of the present invention relates to the medical use of the GLP-1 / GIP coagonists described herein.

[0016] In one aspect, the invention relates to the use of a GLP-1 / GIP coagonist as described herein for the prevention or treatment of diabetes, obesity, and / or liver disease. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] In the following, the Greek letters may be represented by their symbols or by their corresponding names, e.g., α is alpha, β is beta, ε is epsilon, γ is gamma, ω is omega, etc. The Greek letter μ may also be represented by "u", e.g., μl is ul, μM is uM, etc.

[0018] GLP-1 / GIP receptor coagonists The present invention relates to compounds that are GLP-1 receptor agonists and GIP receptor agonists, also referred to as GLP-1 / GIP receptor coagonists or simply coagonists.

[0019] The term "compound" is used herein to refer to a molecular entity, and therefore, a "compound" may have different structural elements other than the minimum elements defined for each compound or group of compounds. As long as the compound contains the defined structural and / or functional elements, it follows that the compound may be a peptide or a derivative thereof.

[0020] The term "compound" is also intended to encompass the pharma- ceutically relevant forms of the compounds herein, i.e., the compounds as defined herein, or a pharma- ceutically acceptable salt, amide, or ester thereof.

[0021] The term "analog" generally refers to a peptide whose sequence has one or more amino acid changes compared to a reference amino acid sequence. An "analog" may include amino acid extensions at the N- and / or C-terminal positions and / or truncations at the N- and / or C-terminal positions.

[0022] In general, amino acid residues can be identified by their full name, their one-letter code, and / or their three-letter code. These three methods are fully equivalent.

[0023] Amino acids are molecules that contain an amino group and a carboxylic acid group, and optionally one or more additional groups often referred to as side chains.

[0024] The term "amino acid" includes not only proteinogenic (or natural) amino acids (particularly the 20 standard amino acids), but also non-proteinogenic (or unnatural) amino acids. Proteinogenic amino acids are those that are naturally incorporated into proteins. Standard amino acids are those that are encoded by the genetic code. Non-proteinogenic amino acids are either not found in proteins or are not produced by standard cellular mechanisms (e.g., they may not be subject to post-translational modification). Non-limiting examples of non-proteinogenic amino acids are Aib (α-aminoisobutyric acid or 2-aminoisobutyric acid), norleucine, norvaline, as well as the D-isomers of the proteinogenic amino acids.

[0025] Hereinafter, each amino acid of a peptide for which the optical isomer is not specified will be understood to mean the L-isomer (unless otherwise specified).

[0026] The GLP-1 / GIP receptor coagonist described herein comprises or consists of the peptide and substituents defined below. In some embodiments, the peptide is a synthetic peptide that is made to optimize activity at the GLP-1 receptor and the GIP receptor. Compounds with suitable receptor binding activity for both the GLP-1 receptor and the GIP receptor are identified as shown in the examples herein.

[0027] The compounds further exhibit extended half-lives through substituents that include fatty acid groups.

[0028] In some embodiments, the carboxy terminus of the peptide retains a -COOH group. In some embodiments, the compounds may optionally include an amide group (C(=O)-NH2) at the C-terminus, which is a naturally occurring modification that replaces -OH with -NH2 as found in native exendin-4.

[0029] peptide The GLP-1 / GIP receptor coagonists described herein comprise peptides and substituents as defined below, where the substituents are attached to the peptide backbone via amino acid residues.

[0030] In some embodiments, the amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GX 30 X 31 X 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:47) with an optional amide modification at the C-terminus, In the formula, X2 is Aib or A; X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 30 is G or absent, X 31 is P or absent, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent.

[0031] In some embodiments, the amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPX 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:36), with an optional amide modification at the C-terminus; During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent.

[0032] In one embodiment, X 39 is absent. 38 and X 39 is absent. 37 , X 38 , and X 39 is absent. 36 , X 37 , X 38 , and X 39 is absent. 35 , X 36 , X 37 , X 38 , and X 39 is absent. 34 , X 35 , X 36 , X 37 , X 38 , and X 39 is absent. 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 is absent. 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 is absent. 31, X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 is absent. 30 , X 31 , X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 In further such embodiments, X is absent. 32 X 33 X 34 X 35 In further such embodiments, X is SSGA. 32 X 33 X 34 X 35 In further such embodiments, X is ESGA. 32 X 33 X 34 X 35 is SKGA. In a further embodiment thereof, the peptide has an amide modification at the C-terminus.

[0033] In one embodiment, the amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPSSGAPPPS (SEQ ID NO:37) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R.

[0034] In one embodiment, the amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPESGAPPPS (SEQ ID NO:38) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R.

[0035] In one embodiment, the amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28GGPSKGAPPPS (SEQ ID NO: 39) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R.

[0036] In one embodiment, X2 is Aib. In one embodiment, X2 is A. In one embodiment, X6 is F. In one embodiment, X6 is V. In one embodiment, X 12 is I. In one embodiment, X 12 is Y. In one embodiment, X 13 is Y. In one embodiment, X 13 is A. In one embodiment, X 13 is L. In one embodiment, X 13 is I. In one embodiment, X 16 is K. In one embodiment, X 16 E. In one embodiment, X 20 is Q. In one embodiment, X 20 is R. In one embodiment, X 20 is E. In one embodiment, X 20 is H. In one embodiment, X 21 is A. In one embodiment, X 21 is E. In one embodiment, X 23 is I. In one embodiment, X 23 is V. In one embodiment, X 24 is E. In one embodiment, X 24is Q. In one embodiment, X 24 is N. In one embodiment, X 28 is A. In one embodiment, X 28 is R. In one embodiment, X 30 is G. In one embodiment, X 31 is P.

[0037] In one embodiment, X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 is selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE, and LLEEQAAREFIE. 13 LEX 16 QAAX 20 X 21 FX 23 X 24 In one embodiment, X is 13 LEX 16 QAAX 20 X 21 FX 23 X 24 In one embodiment, X is 13 LEX 16 QAAX 20 X 21 FX 23 X 24 In one embodiment, X is 13 LEX 16 QAAX 20 X 21 FX 23 X 24 is LLEEQAAREFIE.

[0038] In further embodiments, the amino acid sequence of the peptide is any one of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 33, 34, and 35.

[0039] In one embodiment, the amino acid sequence of the peptide is any one of SEQ ID NOs:10, 22, or 25.

[0040] In one embodiment, the amino acid sequence of the peptide is SEQ ID NO:25.

[0041] In one embodiment, the amino acid sequence of the peptide is any one of SEQ ID NOs: 18, 20, 23, 24.

[0042] In one embodiment, the amino acid sequence of the peptide is SEQ ID NO:33, 34, or 35.

[0043] In further such embodiments, the peptide has an amide modification at the C-terminus.

[0044] derivative In some embodiments, the GLP-1 receptor agonist and the GIP receptor agonist comprise or consist of a substituent, as defined below, covalently attached to the peptide.

[0045] Such compounds may be called derivatives of the peptide since they are obtained by covalently attaching substituents to the peptide backbone.

[0046] One aspect of the invention is a compound comprising a peptide and a substituent, wherein the amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GX 30 X 31 X 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:47) with an optional amide modification at the C-terminus, In the formula, X2 is Aib or A; X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 30 is G or absent, X 31 is P or absent, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent, a compound, wherein the substituent is attached to the peptide via a lysine (K) at position 16 or 33; or a pharma- ceutically acceptable salt thereof.

[0047] One embodiment of the present invention is a compound comprising a peptide and a substituent, wherein the amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPX 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:36) with an optional amide modification at the C-terminus, In the formula, X2 is Aib or A; X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent, a compound, wherein the substituent is attached to the peptide via a lysine (K) at position 16 or 33; or a pharma- ceutically acceptable salt thereof.

[0048] In further embodiments, the peptide may be defined as described herein above.

[0049] substituent In one embodiment, a substituent described herein is attached to a peptide described herein via a lysine (K) residue at position 16 or 33.

[0050] In one embodiment, the substituent is attached to the peptide via the epsilon-amino group of a lysine (K) when the lysine is included at position 16 or 33.

[0051] In one embodiment, the substituent is a chemical structure covalently attached to the peptide that can form a non-covalent complex with plasma albumin and therefore promote circulation of the co-agonist in the bloodstream and can also have the effect of prolonging the duration of action of the co-agonist due to the fact that the co-agonist-albumin complex is only slowly removed by renal clearance.

[0052] In one embodiment, the substituent comprises a fatty acid group. In one such embodiment, the fatty acid group comprises a carbon chain comprising at least 8 consecutive -CH2- groups. In one embodiment, the fatty acid group comprises at least 10 consecutive -CH2- groups, e.g., at least 12 consecutive -CH2- groups, at least 14 consecutive -CH2- groups, at least 16 consecutive -CH2- groups, at least 18 consecutive -CH2- groups.

[0053] In one embodiment, the fatty acid group contains 8 to 20 consecutive -CH2-. In one embodiment, the fatty acid group contains 10 to 18 consecutive -CH2-. In one embodiment, the fatty acid group contains 12 to 18 consecutive -CH2-. In one embodiment, the fatty acid group contains 14 to 18 consecutive -CH2-.

[0054] In some embodiments, the substituent is composed of several elements, such as a protractor element and one or more linker elements. In one embodiment, the term "protractor" is used to describe a fatty acid group that is the terminal portion of the substituent that is responsible for extending the half-life of the compound.

[0055] In one embodiment, the protractor (Prot) is Chemical formula 1: HOOC-(CH2) n -CO-*, where n is an integer ranging from 8 to 20, and may be referred to as C(n+2) diacids; or Chemical formula 1b: [ka] (wherein n is an integer in the range of 8 to 20).

[0056] In one embodiment, the substituent further comprises one or more linker elements. In some embodiments, the linker elements are linked to each other and to the protractor by amide bonds and are referred to as "Z" (see further below).

[0057] As further defined herein below, the number of linker elements is up to three and can be referred to as -Z1-Z2-Z3-, where Z1 is linked to a protractor (Prot) and the last Z element is linked to a peptide, in which case the substituent can be referred to as Prot-Z1-Z2-Z3-. Thus, the symbol * above indicates the point of attachment to Z1, which is nitrogen when attached via an amide bond. In one embodiment, when Z1 is a bond (see below), the symbol * indicates the point of attachment of the adjacent Z element to the nitrogen.

[0058] In one embodiment, the substituents are defined by Prot-Z1-Z2-Z3-, where Prot- is selected from Formula 1, Formula 1b, where n is an integer ranging from 16 to 20.

[0059] In certain embodiments, n in Formula 1 or Formula 1b is 14, 15, 16, 17, 18, 19, or 20.

[0060] In certain embodiments, n in formula 1 or formula 1b is 14, 15, 16, 17, or 18.

[0061] In certain embodiments, n in formula 1 or formula 1b is 14, 16, or 18.

[0062] In certain embodiments, n in Formula 1 or Formula 1b is 16, 17, 18, 19, or 20.

[0063] In certain embodiments, n in Formula 1 or Formula 1b is 16, 18, or 20.

[0064] In certain embodiments, n in formula 1 or formula 1b is 18 or 20.

[0065] In certain embodiments, the protractor (Prot) is a C16 diacid or a C18 diacid.

[0066] In certain embodiments, the protractor (Prot) is a C18 diacid or a C20 diacid.

[0067] In certain embodiments, the protractor (Prot) is a C16 diacid, a C18 diacid, or a C20 diacid.

[0068] The term "bond" as used herein means a covalent bond. When a linker element Z1-Z3 is defined as a bond, it is equivalent to the situation where that linker element is absent. Any indication herein below that any of Z1-Z3 is a bond may be understood to mean that any of Z1-Z3 is absent, such that the previous Z element is covalently linked to the next Z element that is not a "bond" (or is not absent).

[0069] In some embodiments, linker elements Z1-Z3 are individually selected from chemical moieties capable of forming an amide bond, including amino acid-like moieties such as Glu, γGlu (also called gamma Glu or gGlu and defined by *-NH-CH-(COOH)-CH2-CH2-CO-*), ε-Lys (also called epsilon Lys or eLys and defined by *-NH-(CH2)4-CH(NH2)-CO-*), Ser, Ala, Thr, Ado, Aeep, and Aeeep, as well as additional moieties described below.

[0070] In one embodiment, Z1 is selected from γGlu, Glu, or a bond.

[0071] In one embodiment, Z1 is γGlu.

[0072] In one embodiment, Z2 and Z3 are independently selected from Glu, ε-Lys, γGlu, Gly, Ser, Ala, Thr, Ado, Aeep, Aeeep, and a bond.

[0073] Glu, Gly, Ser, Ala, and Thr are amino acid residues well known in the art.

[0074] ε-Lys is defined by the chemical formula 2: -NH-(CH2)4-CH(NH2)-CO- It may be represented by chemical formula 2b: [ka] .

[0075] γGlu is defined by the chemical formula 3: -NH-CH(COOH)-(CH2)2-CO- It may be represented by chemical formula 3b: [ka] .

[0076] Ado is defined by the chemical formula 4: -NH-(CH)-O-(CH)-O-CH-CO- and may be referred to as 8-amino-3,6-dioxaoctanoic acid; Chemical formula 4b: [ka] It may be expressed as:

[0077] Aeep is defined by the chemical formula 5: *NH-CH2CH2OCH2CH2OCH2CH2CO* Chemical formula 5b: [ka] It may be expressed as:

[0078] Aeeep is defined by the chemical formula 6: *NH-CH2CH2OCH2CH2OCH2CH2OCH2CH2CO* Chemical formula 6b: [ka] It may be expressed as:

[0079] In one embodiment, Z2 and Z3 are independently selected from Glu, ε-Lys, γGlu, Gly, Ala, Ado, Aeep, Aeeep, and a bond.

[0080] In one embodiment, Z2 and Z3 are independently selected from Glu, ε-Lys, γGlu, Gly, Ala, Ado, and a bond.

[0081] In one embodiment, Z2 and Z3 are independently selected from Glu, ε-Lys, γGlu, Gly, Ado, and a bond.

[0082] In one embodiment, Z2 and Z3 are independently selected from ε-Lys, γGlu, Gly, Ado, and a bond.

[0083] In one embodiment, Z2 and Z3 are independently selected from ε-Lys, γGlu, Ado, and a bond.

[0084] In one embodiment, Z2 and Z3 are ε-Lys or Ado.

[0085] In one embodiment, Z2 and Z3 are Ado.

[0086] In one embodiment, Z2 and Z3 are ε-Lys.

[0087] In one embodiment, the substituents are selected from substituents A, B, C, D, and E, which are defined as follows:

[0088] In one embodiment, the substituents are selected from substituents A, B, and C, which are defined as follows: [Table 1]

[0089] In some embodiments, the substituent is covalently attached to the lysine residue of the coagonist by acylation, i.e., via an amide bond formed between the carboxylic acid group of the substituent and the epsilon-amino group of the lysine residue.

[0090] In one embodiment, the substituent is covalently attached to the lysine residue at position 16 of the peptide backbone by acylation, i.e., via an amide bond formed between the carboxylic acid group of the substituent and the epsilon-amino group of the lysine residue.

[0091] In one embodiment, the substituent is covalently attached to the lysine residue at position 33 of the peptide backbone by acylation, i.e., via an amide bond formed between the carboxylic acid group of the substituent and the epsilon-amino group of the lysine residue.

[0092] The co-agonists may exist in different stereoisomeric forms that have the same molecular formula and arrangement of bonded atoms, but differ only in the three-dimensional orientation of their atoms in space. The stereoisomerism of the exemplified co-agonists is indicated in the experimental section, names, and structures using standard nomenclature. Unless otherwise specified, the present invention relates to all stereoisomeric forms of the embodied derivatives.

[0093] Functional receptor activation activity The functional activity of the GLP-1 / GIP receptor agonists described herein can be tested in vitro as described in Example 2 herein.

[0094] 50% effective concentration (EC 50 The term EC (Estimated Effectiveness) generally refers to the concentration that elicits a response halfway between the baseline and maximum by reference to a dose-response curve. 50 is used as a measure of a compound's potency and represents the concentration at which 50% of its maximal effect is observed.

[0095] Thus, the in vitro potency of a compound can be determined as described herein, and EC 50 The EC 50 The lower the , the better the efficacy.

[0096] To characterize such compounds, it may further be relevant to consider the in vitro potency relative to the native hormone for each receptor.

[0097] In vitro potency may be determined, for example, in culture medium containing membranes expressing the appropriate GLP-1 and / or GIP receptors and / or in assays using whole cells expressing the appropriate GLP-1 and / or GIP receptors.

[0098] For example, the functional response of human GLP-1 and / or GIP receptors can be measured in a receptor gene assay, for example in a stably transfected BHK cell line expressing human GLP-1 and / or GIP receptors and containing DNA of cAMP response element (CRE) and gene of firefly luciferase (CRE luciferase) linked to a promoter. The production of cAMP as a result of activation of GLP-1 and / or GIP receptors then leads to the expression of luciferase. Luciferase can be determined by adding luciferin, which is enzymatically converted to oxyluciferin to produce bioluminescence, which is measured as a reporter of in vitro potency. An example of such an assay is described in Example 2 herein. It is also important to note that receptor activity can be affected by the presence or absence of human serum albumin (HSA) in the assay medium, since the compounds may contain substituents designed to bind to albumin. EC in the absence of HSA 50 Compared to EC 50 A decrease in the potency of the compound in the presence of HSA, as indicated by an increase in , indicates an interaction of the compound with HSA and predicts a prolonged duration of action in vivo.

[0099] In one embodiment, the compounds have potent in vitro effects in activating human GLP-1 and GIP receptors.

[0100] In one embodiment, the compound has an EC of less than 20 pM in the CRE luciferase reporter assay described in Example 2 herein when performed without HSA. 50 can activate human GLP-1 and GIP receptors in vitro.

[0101] In one embodiment, the compounds have an EC of 100 pM or less, more preferably less than 50 pM, or most preferably less than 20 pM. 50 The compounds have in vitro potencies at the human GLP-1 receptor and GIP receptor as determined using the methods of Example 2 corresponding to:

[0102] In one embodiment, the EC 50 is 1 to 25 pM, such as 1 to 20 pM, such as 1 to 15 pM, or such as 1 to 10 pM.

[0103] Pharmacokinetic properties The pharmacokinetic properties of the co-agonist compounds can be further determined in vivo by pharmacokinetic (PK) studies. This characterization can be performed using animal models such as mice, rats, monkeys, dogs, or pigs.

[0104] In such studies, animals are typically administered a single dose of drug in a relevant formulation, either intravenously, subcutaneously (sc) or orally (po). Blood samples are taken at predefined times after dosing and analyzed for drug concentration using a relevant quantitative assay. Based on these measurements, the time-plasma concentration profile of the test compound is plotted and a so-called non-compartmental pharmacokinetic analysis of the data is performed. The terminal half-life is an important parameter, since a long half-life indicates that a compound may be able to be administered less frequently. The in vivo terminal half-life (t1 / 2) can be measured using a suitable model, for example after intravenous administration in minipigs as described in Example 3 or after oral administration in dogs as described in Example 4.

[0105] In one embodiment, the terminal half-life is the in vivo half-life (t1 / 2) in minipigs following intravenous administration, for example, as described in Example 3 herein.

[0106] In one embodiment, the terminal half-life in the minipig is at least 24 hours, such as at least 30 hours, or at least 40 hours.

[0107] In one embodiment, the terminal half-life is the in vivo half-life (t1 / 2) in dogs following oral administration, for example, as described in Example 4 herein.

[0108] In one embodiment, the terminal half-life in dogs is at least 24 hours, such as at least 40 hours, or such as at least 50 hours.

[0109] Pharmaceutically acceptable salts In some embodiments, the co-agonists described herein are in the form of pharma- ceutically acceptable salts. Salts are formed, for example, by chemical reactions between bases and acids, such as 2NH3+H2SO4→(NH4)2SO4. Salts may be basic or acidic salts, or neither (i.e., neutral salts). In water, basic salts generate hydroxide ions, and acidic salts generate hydronium ions. Salts of the co-agonists may be formed by adding cations or anions between anionic or cationic groups, respectively. These groups may be located on the peptide and / or on the substituents of the derivative. Non-limiting examples of anionic groups include any free carboxylic acid groups in the substituents, if present, as well as free carboxylic acid groups in the peptide. The peptide may include a free carboxylic acid group at the C-terminus, if present, as well as any free carboxyl groups of internal acidic amino acid residues, such as Asp and Glu.

[0110] Non-limiting examples of cationic groups include any free amino groups in a substituent, if present, and free amino groups in a peptide. A peptide may contain a free amino group at the N-terminus, if present, and any free imidazole or amino groups of internal basic amino acid residues such as His, Arg, and Lys.

[0111] In certain embodiments, the peptide or derivative is in the form of a pharma- ceutically acceptable salt.

[0112] Generation Process Coagonists may be produced by classical peptide synthesis, e.g., solid phase peptide synthesis, using, for example, t-Boc or Fmoc chemistry or other well-established techniques, see, for example, Greene and Wuts, "Protective Groups in Organic Synthesis", John Wiley & Sons, 1999, Florencio Zaragoza Dorwald, "Organic Synthesis on Solid Phase", Wiley-VCH Verlag GmbH, 2000, and "Fmoc Solid Phase Peptide Synthesis", Edited by WCChan and PDWhite, Oxford University Press, 2000.

[0113] Alternatively, the compounds may be produced by recombinant methods, for example by culturing a host cell that contains a DNA sequence encoding the peptide sequence and is capable of expressing the peptide in a suitable nutrient medium under conditions that permit expression of the peptide. Non-limiting examples of host cells suitable for expression of these peptides are Escherichia coli, Saccharomyces cerevisiae, and mammalian BHK or CHO cell lines.

[0114] Coagonists containing unnatural amino acids and / or covalently attached substituents may be generated as described in the experimental part.

[0115] Specific examples of how to prepare some co-agonists are included in the experimental part.

[0116] A further aspect of the present invention relates to methods for preparing the peptides described herein.

[0117] A further aspect of the present invention relates to methods for preparing the GLP-1 / GIP coagonists described herein.

[0118] In one embodiment, the method for preparing the compounds described herein comprises a step of solid phase peptide synthesis. Substituents may be built sequentially as part of the solid phase peptide synthesis or may be generated separately and attached via lysine residues after peptide synthesis.

[0119] In one embodiment, the compounds are produced by a two-step process in which a substituent is attached to one of two peptide fragments, which are then ligated.

[0120] Pharmaceutical Compositions In a further aspect, the present invention relates to a pharmaceutical composition comprising a GLP-1 / GIP receptor coagonist as described herein. Compositions comprising a compound of the present invention or a pharma- ceutically acceptable salt thereof, and optionally one or more pharma- ceutically acceptable excipients, can be prepared as known in the art.

[0121] The term "excipient" broadly refers to any ingredient other than the active therapeutic ingredient. An excipient can be an inert, inactive, and / or medically inactive substance. Excipients can serve a variety of purposes, for example, as carriers, vehicles, fillers, binders, lubricants, glidants, disintegrants, flow control agents, crystallization inhibitors, solubilizers, stabilizers, colorants, flavoring agents, surfactants, emulsifiers, or combinations thereof, and / or to improve administration and / or to improve absorption of the active agent. The amount of each of the excipients used can vary within ranges conventional in the art. Techniques and excipients that can be used to formulate oral dosage forms are described in the Handbook of Pharmaceutical Excipients (e.g., 8th ed., Sheskey et al., Eds., American Pharmaceuticals Association and Pharmaceutical Press, publications department of the Royal Pharmaceutical Society of Great Britain (2017), and any subsequent editions), and Remington: The Science and Practice of Pharmacy (e.g., 22nd ed., Remington and Allen, Eds., Pharmaceutical Press (2013), and any subsequent editions).

[0122] In one embodiment, the pharmaceutical composition may be a solid formulation, such as a freeze-dried or spray-dried composition, which may be used as is or may be used by the physician or patient after adding a solvent and / or diluent before use.In a further embodiment, the pharmaceutical composition may be a solid formulation consisting of the active ingredient, N-[8-(2-hydroxybenzoyl)amino]caprylate, and one or more additional excipients known in the art, for example, using any one or more of the formulations described in WO2012 / 080471, WO2013 / 189988, or WO2019 / 149880.

[0123] Alternatively, the pharmaceutical composition is a liquid formulation, such as an aqueous formulation. Liquid compositions suitable for injection can be prepared using conventional techniques in the pharmaceutical industry, which involve dissolving and mixing the components as necessary to produce the desired final product. Thus, according to a procedure, the GLP-1 / GIP coagonist described herein is dissolved in a suitable buffer at a suitable pH. The composition can be sterilized, for example, by sterile filtration.

[0124] Pharmaceutical indications A further aspect of the present invention relates to the use of a GLP-1 / GIP receptor coagonist compound as described herein as a medicament.

[0125] In one embodiment, the compounds described herein are for use in the medical treatment of: (i) prevention and / or treatment of all forms of diabetes including hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, non-insulin dependent diabetes, MODY (maturity onset diabetes of the young), gestational diabetes, and / or reduction of HbA1C; (ii) delaying or preventing the progression of diabetic disease, such as the progression of type 2 diabetes, delaying the progression from impaired glucose tolerance (IGT) to insulin-requiring type 2 diabetes, delaying or preventing insulin resistance, and / or delaying the progression from insulin-non-requiring type 2 diabetes to insulin-requiring type 2 diabetes; (iii) prevention and / or treatment of eating disorders such as obesity, for example by reducing food intake, weight loss, suppressing appetite, inducing satiety; treatment or prevention of binge eating disorder, bulimia nervosa, and / or obesity induced by administration of antipsychotics or steroids, slowing gastric emptying, increasing physical mobility, and / or preventing and / or treating comorbidities of obesity such as osteoarthritis and / or urinary incontinence; (iv) weight maintenance after successful weight loss (drug-induced or diet and exercise-induced), i.e., prevention of weight gain after successful weight loss. (v) prevention and / or treatment of liver disorders such as hepatic steatosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver inflammation, or fatty liver.

[0126] In one embodiment the compound is for use in a method for the treatment and / or prevention of diabetes and / or obesity.

[0127] In one embodiment, the compound is for use in a method for treating diabetes and / or obesity.

[0128] In one embodiment the compound is for use in a method for treating and / or preventing type 2 diabetes.

[0129] In one embodiment, the compound is for use in a method for treating type 2 diabetes.

[0130] In one embodiment the compound is for use in a method for the treatment and / or prevention of obesity.

[0131] In one embodiment, the compound is for use in a method for treating obesity.

[0132] In one embodiment, the compound is for use in a method for controlling weight. In one embodiment, the compound is for use in a method for reducing appetite. In one embodiment, the compound is for use in a method for reducing food intake.

[0133] Embodiment 1. A compound comprising a peptide and a substituent, the amino acid sequence of the peptide being: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24WLLX 28 GX 30 X 31 X 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:47) with an optional amide modification at the C-terminus, In the formula, X2 is Aib or A; X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 30 is G or absent, X 31 is P or absent, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent, the substituent is attached to the peptide via a lysine (K) at position 16 or 33; or a pharma- ceutically acceptable salt thereof. 2. A peptide and a substituent, the peptide having an amino acid sequence as follows: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPX 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:36) with an optional amide modification at the C-terminus, In the formula, X2 is Aib or A; X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38is P or absent, X 39 is S or absent, The compound of embodiment 1, wherein the substituent is attached to the peptide via a lysine (K) at position 16 or 33. or a pharma- ceutically acceptable salt thereof. 3.X 36 , X 37 , X 38 , and X 39 The compound of embodiment 1, wherein 4.X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 The compound of embodiment 1, wherein 5.X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 The compound of embodiment 1, wherein 6.X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 The compound of embodiment 1, wherein 7. The compound of any one of embodiments 1-6, wherein the peptide has the amide modification at the C-terminus. 8.X 32 X 33 X 34 X 35 The compound of any one of embodiments 1, 2, or 7, wherein is selected from the group consisting of SSGA, ESGA, and SKGA. 9.X 32 X 33 X 34 X 35is a SSGA, a compound according to any one of embodiments 1, 2, or 7. 10.X 32 X 33 X 34 X 35 The compound of any one of embodiments 1, 2, or 7, wherein is ESGA. 11.X 32 X 33 X 34 X 35 The compound of any one of embodiments 1, 2, or 7, wherein is SKGA. 12. The amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPSSGAPPPS (SEQ ID NO:37) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R. 13. The amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X24 WLLX 28 GGPESGAPPPS (SEQ ID NO:38) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R. 14. The amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPSKGAPPPS (SEQ ID NO: 39) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28is A or R. 15.X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 The compound of any one of embodiments 1-14, wherein is selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE, and LLEEQAAREFIE. 16. The compound according to embodiment 1, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35. 17. The compound of embodiment 16, wherein the peptide has the amide modification at the C-terminus. 18. The compound has an EC of less than 20 pM when measured without HSA in the CRE luciferase receptor assay as described in Example 2. 50 The compound according to any one of embodiments 1 to 17, which activates human GLP-1 and GIP receptors in vitro. 19. The compound of any one of embodiments 1-18, wherein the compound has a half-life of at least 35 hours in minipigs. 20. The compound according to any one of the preceding embodiments, wherein the substituent is attached via 16Lys. 21. The compound according to any one of embodiments 1-20, wherein the substituent is attached via 33Lys. 22. The compound according to any one of embodiments 1-21, wherein the substituent comprises at least one protractor. 23. The compound according to embodiment 22, wherein the protractor is a fatty acid group. 24. The protractor has the formula 1: HOOC-(CH2) n The compound of embodiment 23, which is a diacid defined by -CO-, where n is an integer in the range of 8 to 20, e.g., n=14, 16, or 18. 25. The protractor has the formula 1: HOOC-(CH2) nThe compound of embodiment 23, which is a diacid defined by -CO-, where n is an integer in the range of 8 to 20, e.g., n=16 or 18. 26. The compound according to any one of the preceding embodiments, wherein the substituent comprises at least one linker element. 27. The compound according to embodiment 26, wherein the substituent comprises up to three linker elements. 28. The compound according to embodiment 27, wherein the substituent comprises up to three linker elements, designated -Z1-Z2-Z3-, where -Z1- is linked to the protractor and -Z3- is linked to the peptide. 29. The substituent is: Prot-Z1-Z2-Z3- (In the formula, Prot is a C16-C20 diacid; Z1 is γGlu or a bond; Z2 is εLys, γGlu, or Ado; The compound of any one of embodiments 1-28, wherein Z3 is εLys or Ado. 30. The compound according to embodiment 29, wherein Z1- is -γGlu-. 31. The compound according to embodiment 29 or 30, wherein -Z2-Z3- and are -Ado-Ado-. 32. The compound according to embodiment 29, 30, or 31, wherein -Z2-Z3- and are -εLys-εLys-. 33. The compound of any one of embodiments 1-17, wherein the substituents are selected from the group consisting of: [ka] 34. The compound of any one of embodiments 1-17, wherein the substituents are selected from the group consisting of: [ka] 35. The compound according to any one of embodiments 1-17, wherein the substituent is attached to the peptide via the epsilon-amino group of a lysine (K). 36. The compound of embodiment 1, wherein the compound is selected from the group consisting of: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] 37. The compound of embodiment 1, wherein the compound is selected from the group consisting of: [ka] [ka] 38. The compound of embodiment 1, wherein the compound is selected from the group consisting of: [ka] [ka] 39. The compound of embodiment 1, wherein the compound is: [ka] 40. The compound of embodiment 1, wherein the compound is: [ka] 41. A compound according to any one of embodiments 1 to 40 for use as a medicament. 42. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 40. 43. The composition of embodiment 42, wherein the composition is an aqueous liquid. 44. The composition of embodiment 42, wherein the composition is a solid composition. 45. A pharmaceutical composition according to any one of embodiments 42 to 44, for the prevention and / or treatment of diabetes and / or obesity. 46. ​​The pharmaceutical composition according to any one of embodiments 42 to 44 for the prevention and / or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH) liver inflammation, and / or fatty liver. 47. A method for preventing and / or treating diabetes and / or obesity, comprising administering to a patient a pharma- ceutically active amount of a compound according to any one of embodiments 1 to 40. 48. A method for preventing and / or treating liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH) liver inflammation, and / or fatty liver, comprising administering to a patient a pharma- ceutical active amount of a compound according to any one of embodiments 1-40. 49. A peptide, the amino acid sequence of which is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GX 30 X 31 X32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:47) with an optional amide modification at the C-terminus, During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 30 is G or absent, X 31 is P or absent, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent. 50. A peptide, the amino acid sequence of which is: YX2EGTX6TSDYSX 12 X13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPX 32 X 33 X 34 X 35 X 36 X 37 X 38 X 39 (SEQ ID NO:36) with an optional amide modification at the C-terminus, During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R, X 32 is E, S, or absent, X 33 is S, K, or absent, X 34 is G or absent, X 35 is A or absent, X 36 is P or absent, X 37 is P or absent, X 38 is P or absent, X 39 is S or absent. 51.X 36 , X 37 , X38 , and X 39 50. The peptide of embodiment 49, wherein: 52.X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 50. The peptide of embodiment 49, wherein: 53.X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 50. The peptide of embodiment 49, wherein: 54.X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , X 36 , X 37 , X 38 , and X 39 50. The peptide of embodiment 49, wherein: 55.X 32 X 33 X 34 X 35 The peptide according to embodiment 49 or 50, wherein is selected from the group: SSGA, ESGA, and SKGA. 56.X 32 X 33 X 34 X 35 SSGA, a peptide according to embodiment 55. 57.X 32 X 33 X 34 X 35 is ESGA. 58.X 32 X 33 X 34 X 35 is SKGA. 59. The peptide of any one of embodiments 47 to 58, wherein the peptide has the amide modification at the C-terminus. 60. The amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPSSGAPPPS (SEQ ID NO:37) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R. 61. The amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPESGAPPPS (SEQ ID NO:38) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R. 62. The amino acid sequence of the peptide is: YX2EGTX6TSDYSX 12 X 13 LEX 16 QAAX 20 X 21 FX 23 X 24 WLLX 28 GGPSKGAPPPS (SEQ ID NO: 39) During the ceremony, X2 is Aib or A, X6 is F or V, X 12 is I or Y, X 13 is Y, A, L, or I; X 16 is K or E, X 20 are Q, R, E, and H, X 21 is A or E, X 23 is I or V, X 24 is E, Q, or N, X 28 is A or R. 63. The peptide according to embodiment 49, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35. 64. The peptide of embodiment 63, wherein the peptide has the amide modification at the C-terminus. 65.X 13 LEX 16 QAAX 20 X 21 FX 23 X24 65. The peptide of any one of embodiments 49-64, wherein is selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE, and LLEEQAAREFIE. 66. The peptide has an EC of less than 20 pM when measured without HSA in a CRE luciferase receptor assay as described in Example 2. 50 65. The peptide according to any one of embodiments 49 to 64, which activates human GLP-1 and GIP receptors in vitro. 67.X 16 The peptide of any one of embodiments 49-64, wherein 68.X 33 The peptide of any one of embodiments 49-64, wherein 69. The peptide of embodiment 49, wherein the amino acid sequence of the peptide is any of SEQ ID NOs: 15, 28, 29, 31, 32, or 43. 70. The peptide of embodiment 49, wherein the amino acid sequence of the peptide is SEQ ID NO: 31. 71. A process for preparing a compound according to any one of embodiments 1 to 40. 72. A method for preparing a peptide according to any one of embodiments 49 to 64. 73. A method for preparing a peptide according to any one of embodiments 49 to 70.

[0134] Methods and Examples List of Abbreviations The following abbreviations are in alphabetical order and will be used below. Ac: Acetyl Ado (also called OEG): 8-amino-3,6-dioxaoctanoic acid Aib: α-aminoisobutyric acid API: Active Pharmaceutical Ingredient AUC: Area under the curve BG: Blood sugar BHK: Baby Hamster Kidney Boc: tert-butyloxycarbonyl BW: Weight CAS: Chemical Abstracts Service Cl-HOBt: 6-chloro-1-hydroxybenzotriazole DCM: dichloromethane DIC: Diisopropylcarbodiimide DIPEA: N,N-diisopropylethylamine DMEM: Dulbecco's modified Eagle's medium DPBS: Dulbecco's phosphate buffered saline EDTA: Ethylenediaminetetraacetic acid ELISA: Enzyme-linked immunosorbent assay equiv: molar equivalent FBS: Fetal bovine serum Fmoc: 9-fluorenylmethyloxycarbonyl GIP: glucose-dependent insulinotropic polypeptide GIPR: glucose-dependent insulinotropic polypeptide receptor GLP-1: Glucagon-like peptide 1 GLP-1R: Glucagon-like peptide 1 receptor h: time HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid HFIP: 1,1,1,3,3,3-Hexafluoro-2-propanol or Hexafluoroisopropanol HPLC: High-performance liquid chromatography HSA: human serum albumin ip: intraperitoneal IPGTT: Intraperitoneal glucose tolerance test iv: intravenous LCMS: Liquid Chromatography Mass Spectrometry LYD: Landrace Yorkshire Duroc MeCN: Acetonitrile MeOH: Methanol mM: millimolar mmol: millimolar min:minutes Mtt: 4-methyltrityl MW: molecular weight nM: nanomolar NMP: 1-methyl-pyrrolidin-2-one OEG: 8-amino-3,6-dioxaoctanoic acid (also called Ado) OtBu: tert-butyl ester Oxyma Pure®: Cyano-hydroxyimino-acetic acid ethyl ester Pbf: 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl PBS: Phosphate-buffered saline PD: Pharmacodynamic PK: Pharmacokinetics pM: picomolar RP: Reverse phase RP-HPLC: Reversed-phase high-performance liquid chromatography rpm: rounds per minute RT: room temperature Rt: retention time sc: subcutaneous SD: standard deviation SEC-HPLC: Size Exclusion High Performance Liquid Chromatography SEM: standard error SNAC: Sodium N-[8-(2-hydroxybenzoyl)amino]caprylate SPPS: Solid Phase Peptide Synthesis tBu: tert-butyl TFA: Trifluoroacetic acid TIS: Triisopropylsilane Trt: triphenylmethyl or trityl Trx: tranexamic acid

[0135] General preparation method The methods for solid phase peptide synthesis (including the SPPS method, the deprotection of the amino acids, the cleavage of the peptide from the resin, and its purification) as well as the detection and characterization of the resulting peptides (LCMS method) are described below.

[0136] The resin used for the preparation of C-terminal peptide amides was H-Rink Amide-ChemMatrix resin (e.g., loading 0.5 mmol / g). Unless otherwise specified, the Fmoc-protected amino acid derivatives used were, for example, Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Ile ... The recommended standards were Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Met-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, Fmoc-Lys(Mtt)-OH, and Fmoc-Aib-OH. If nothing else is specified, the natural L-form of the amino acid is used. The alpha-amino group of the N-terminal amino acid was Boc-protected either by using a reagent with a pre-introduced Boc group (e.g., Boc-Tyr(tBu)-OH for peptides with Tyr at the N-terminus) or by exchanging the N-terminal Fmoc protecting group for a Boc protecting group after incorporation of the amino acid at the peptide N-terminus.

[0137] For attachment of modular albumin binding moieties using SPPS, suitably protected building blocks were used, such as, but not limited to, Fmoc-8-amino-3,6-dioxaoctanoic acid (Fmoc-Ado-OH), Boc-Lys(Fmoc)-OH, Fmoc-Glu-OtBu, hexadecanedioic acid mono-tert-butyl ester, octadecanedioic acid mono-tert-butyl ester, nonadecanedioic acid mono-tert-butyl ester, eicosanedioic acid mono-tert-butyl ester, tetradecanedioic acid mono-tert-butyl ester, or 4-(9-carboxynonyloxy)benzoic acid tert-butyl ester. All operations described below were carried out within the synthesis scale range of 0.1-0.2 mmol.

[0138] 1. Synthesis of resin-bound protected peptide scaffold: Method: SPPS_A SPPS was performed using Fmoc-based chemistry on a Protein Technologies SymphonyX solid-phase peptide synthesizer using slight modifications to the protocol provided by the manufacturer. Mixing was achieved by occasional nitrogen bubbling. A stepwise assembly was performed using the following steps: 1) pre-swelling the resin in DMF, 2) deprotecting the Fmoc with two treatments (10 min each) using 20% ​​(v / v) piperidine in DMF, 3) washing with DMF to remove piperidine, 4) coupling the Fmoc-amino acid by adding Fmoc-amino acid (12 eq.) and Oxyma Pure® (12 eq.) each as a 0.6 M solution in DMF, followed by DIC (12 eq.) as a 1.2 M solution in DMF, followed by DMF to reduce the final concentration of each component to 0.3 M, followed by mixing for 0.5-4 h, 4) washing with DMF to remove excess reagent, and 5) a final wash with DCM upon completion of the assembly. Some amino acids, such as but not limited to the sterically hindered amino acid (e.g., Aib) were coupled with extended reaction times (e.g., 4 h) to ensure reaction completion.

[0139] Method: SPPS_B The protected peptidyl resin was synthesized according to the Fmoc method on an Applied Biosystems 431A solid phase peptide synthesizer using the generic Fmoc protocol provided by the manufacturer. Mixing was achieved by vortexing and occasional nitrogen bubbling. Stepwise build-up was performed using the following steps: 1) Activation of the Fmoc-amino acid by dissolving solid Fmoc-amino acid (10 eq.) in Cl-HOBt (10 eq.) as a 1M NMP solution, followed by addition of DIC (10 eq.) as a 1M NMP solution, followed by simultaneous mixing with steps 2-3, 2) Deprotection of the Fmoc using 20% ​​(v / v) piperidine in NMP for the first treatment (3 min) followed by a second treatment (15 min), 3) Washing with NMP to remove piperidine, 4) Addition of the activated Fmoc-amino acid solution to the resin followed by mixing for 45-90 min, 4) Washing with NMP to remove excess reagent, and 5) Final wash with DCM upon completion of build-up. The above standard protected amino acid derivatives were supplied in pre-weighed cartridges (e.g., from Midwest Biotech) and non-standard derivatives were weighed manually. Some amino acids, including but not limited to those following a sterically hindered amino acid (e.g., Aib), were "double coupled" to ensure reaction completion by draining the resin after the first coupling (e.g., 45 min), adding additional reagents (Fmoc-amino acid, DIC, Cl-HOBt), and reacting the mixture again (e.g., 45 min).

[0140] Method: SPPS_C SPPS was performed using Fmoc-based chemistry on a PreludeX solid-phase peptide synthesizer with minor modifications to the protocol provided by the manufacturer. Mixing was achieved by shaking at 350 rpm and occasional nitrogen bubbling. A stepwise build was performed using the following steps: 1) pre-swelling the resin in DMF, 2) Fmoc deprotection in one treatment (3 min at 70° C.) using 20% ​​(v / v) piperidine in DMF, 3) piperidine removal by washing with DMF, 4) coupling of Fmoc-amino acid by adding Fmoc-amino acid (12 eq.) and Oxyma Pure® (12 eq.), each as a 0.4 M solution in DMF, followed by DIC (12 eq.) as a 1.2 M solution in DMF, followed by mixing for 5 min at 70° C., 4) DMF wash to remove excess reagents, and 5) final wash with DCM upon build completion. Some amino acids, including but not limited to those following a sterically hindered amino acid (eg, Aib), were coupled with extended reaction times (eg, 15 min) to ensure reaction completion.

[0141] 2. Attachment of Substituents to the Resin-Bound Protected Peptide Backbone Method:SC_A The N-epsilon-lysine-protected Mtt protecting group was removed by washing the resin with 30% HFIP in DCM for two treatments (45 min each), followed by washing with DCM and DMF. Acylation was performed on a Protein Technologies SymphonyX solid phase peptide synthesizer using the protocol described in method SPPS_A using stepwise addition of building blocks such as, but not limited to, Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-Glu-OtBu, hexadecanedioic acid mono-tert-butyl ester, octadecanedioic acid mono-tert-butyl ester, and eicosanedioic acid mono-tert-butyl ester.

[0142] Method:SC_B The N-epsilon-lysine-protected Mtt protecting group was removed by washing the resin with 30% HFIP in DCM for two treatments (45 min each), followed by washing with DCM and DMF. Acylation was performed on an Applied Biosystems 431A solid phase peptide synthesizer using the protocol described in method SPPS_B, using stepwise addition of building blocks such as, but not limited to, Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-Glu-OtBu, hexadecanedioic acid mono-tert-butyl ester, octadecanedioic acid mono-tert-butyl ester, and eicosanedioic acid mono-tert-butyl ester.

[0143] Method:SC_C The N-epsilon-lysine-protected Mtt protecting group was removed by washing the resin with 30% HFIP in DCM for two treatments (45 min each), followed by washing with DCM and DMF. Acylation was performed on a Protein Technologies PreludeX solid phase peptide synthesizer using the protocol described in method SPPS_C, using stepwise addition of building blocks such as, but not limited to, Boc-Lys(Fmoc)-OH, Fmoc-8-amino-3,6-dioxaoctanoic acid, Fmoc-Glu-OtBu, hexadecanedioic acid mono-tert-butyl ester, octadecanedioic acid mono-tert-butyl ester, and eicosanedioic acid mono-tert-butyl ester.

[0144] 3. Cleavage and purification of resin-bound peptide: Method: CP_A After completion of the side chain synthesis, the peptidyl resin was washed with DCM, dried, and then treated with TFA / water / TIS (95:2.5:2.5 (v / v / v)) for approximately 2 hours, and then precipitated with diethyl ether. The precipitate was washed with diethyl ether, dissolved in a suitable solvent (e.g., 2:1 water / MeCN) and left until all unstable adducts had decomposed. Purification was performed by reversed-phase preparative HPLC (Waters 2545 binary gradient module, Waters 2489 UV / visible detector, Waters fraction collector III) using a Phenomenex Luna C8(2) column (10 μM particle size, 100 Å pore size, dimensions 250×21.2 mm). Separation of impurities and elution of the product were achieved by increasing gradients of MeCN in 0.1% TFA-containing water. Relevant fractions were checked for identity and purity by analytical LCMS. Fractions containing the pure desired peptide were pooled and lyophilized to give the peptide TFA salt as a white solid.

[0145] 4. Salt exchange from TFA to sodium salt: Method: SX_A Lyophilized peptides isolated from method CP_A were dissolved in an appropriate aqueous buffer (e.g., 4:1 water / MeCN, 0.2 M sodium acetate) to 5-20 mg / mL and adjusted to pH 7-8 with 1 M NaOH as necessary to ensure complete dissolution. The buffer containing the peptide was salt exchanged using a Sep-Pak C18 cartridge (0.5-2 g). The cartridge was first equilibrated with 4 column volumes of isopropanol, then 4 column volumes of MeCN, then 8 column volumes of water. The peptide solution was applied to the cartridge and the flow-through was reapplied to ensure complete retention of the peptide. The cartridge was washed with 4 column volumes of water, then 10 column volumes of a buffer (e.g., pH 7.5) including, but not limited to, NaHCO3, NaOAc, or Na2HPO4. The column was washed with 4 column volumes of water and the peptide was eluted with 5-10 column volumes of 50-80% aqueous MeCN. The eluent containing the peptide was lyophilized to give the peptide sodium salt as a white solid, which was used as is.

[0146] General detection and characterization methods LCMS method: Method: LCMS_A LCMS_A was performed on a setup consisting of an Agilent 1260 Infinity series HPLC system and an Agilent Technologies 6120 Quadrupole MS. Eluents: A: 0.05% TFA in water, B: 0.05% TFA in 9:1 MeCN / water.

[0147] The analysis was performed at room temperature (column temperature 37 C) by injecting an appropriate volume of sample onto the column eluted with a gradient of A and B. Column: Phenomenex Kinetex C8, 2.6 μm, 100 Å, 4.6 x 75 mm. Gradient run time: linear 10-80% B over 10 min at a flow rate of 1.0 mL / min. Detection: Diode array detector set at 214 nm. MS ionization mode: API-ES, positive polarity. MS scan mass range: 500-2000 amu. The most abundant isotope of each m / z is reported.

[0148] Method: LCMS_B LCMS_B was performed on a setup consisting of an Agilent 1260 Infinity series HPLC system and an Agilent Technologies 6120 Quadrupole MS. Eluents: A: 0.05% TFA in water, B: 0.05% TFA in 9:1 MeCN / water.

[0149] The analysis was performed at room temperature (column temperature 37 C) by injecting an appropriate volume of sample onto the column eluted with a gradient of A and B. Column: Phenomenex Kinetex C8, 2.6 μm, 100 Å, 4.6 x 75 mm. Gradient run time: linear 20-100% B over 10 min at a flow rate of 1.0 mL / min. Detection: Diode array detector set at 214 nm. MS ionization mode: API-ES, positive polarity. MS scan mass range: 500-2000 amu. The most abundant isotope of each m / z is reported.

[0150] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those skilled in the art. It is therefore to be understood that the appended embodiments are intended to cover all such modifications and changes that fall within the true spirit of the invention.

[0151] Example 1: Synthesis of Compounds The compounds are as follows written using the single letter amino acid code, except for Aib: Substituents are included after the lysine (K) residue to which they are attached. Compound No. 1 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVNWLLAGGPSSGAPPPS-NH2 [ka] SEQ ID NO:1 with C-terminal amide modification, Substituent: C, Substituent position: K16 Synthesis method: SPPS_A, SC_B, CP_A Calculated molecular weight (average): 4873.5Da LCMS_A: Retention time = 6.0 min, observed value [M+3H] 3+ 1625.4, [M+4H] 4+ 1219.1. Compound No. 2 Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVNWLLAGGPSSGAPPPS-NH2 [ka] SEQ ID NO:2 with C-terminal amide modification, Substituent: C, Substituent position: K16 Synthesis method: SPPS_A, SC_B, CP_A Calculated molecular weight (average): 4923.5Da LCMS_A: Retention time = 6.0 min, observed value [M+3H] 3+ 1641.8, [M+4H] 4+ 1237.5 Compound No. 3 Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFVNWLLAGGPSSGAPPPS-NH2 [ka] SEQ ID NO:2 with C-terminal amide modification, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_B, CP_A Calculated molecular weight (average): 4895.4Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1632.7, [M+4H] 4+ 1224.6 Compound No. 4 Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQAFVNWLLAGGPSSGAPPPS-NH2 [ka] SEQ ID NO:3 with C-terminal amide modification, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_B, CP_A Calculated molecular weight (average): 4837.4Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1613.3, [M+4H] 4+ 1210.1 Compound number 5 Y-Aib-EGTFTSDYSYYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQAFVNWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:3, Substituent: B, Substituent position: K16 Synthesis method: SPPS_B, SC_B, CP_A Calculated molecular weight (average): 4838.4Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1613.6, [M+4H] 4+ 1210.4 Compound No. 6 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVNWLLAGGPSSGAPPPS-NH2 [ka] SEQ ID NO:4 with C-terminal amide modification, Substituent: B, Substituent position: K16 Synthesis method: SPPS_B, SC_B, CP_A Calculated molecular weight (average): 4873.5Da LCMS_A: Retention time = 5.6 min, observed value [M+3H] 3+ 1625.3, [M+4H] 4+ 1219.1 Compound No. 7 Y-Aib-EGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFVNWLLAGGPSSGAPPPS-NH2 [ka] SEQ ID NO:4 with C-terminal amide modification, Substituent: A, Substituent position: K16 Synthesis method: SPPS_B, SC_B, CP_A Calculated molecular weight (average): 4907.4Da LCMS_A: Retention time = 6.3 min, observed value [M+3H] 3+ 1636.5, [M+4H] 4+ 1227.9 Compound No. 8 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-NH2 [ka] SEQ ID NO:5 with C-terminal amide modification, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4887.5Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1629.8, [M+4H] 4+ 1222.8 Compound No. 9 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:6, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_B, CP_A Calculated molecular weight (average): 4903.5Da LCMS_A: Retention time = 5.8 min, observed value [M+3H] 3+ 1635.2, [M+4H] 4+ 1226.8 Compound No. 10 Y-Aib-EGTFTSDYSIYLE-K[2-[2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFIEWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:6, Substituent: A, Substituent position: K16 Synthesis method: SPPS_A, SC_B, CP_A Calculated molecular weight (average): 4937.5Da LCMS_A: Retention time = 6.5 min, observed value [M+3H] 3+ 1646.6, [M+4H] 4+ 1235.1 Compound No. 11 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:5, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4888.5Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1630.2, [M+4H] 4+ 1222.9 Compound No. 12 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPSSGAPPPS-OH [ka] Synthesis method: SPPS_A, SC_C, CP_A SEQ ID NO:7, Substituent: B, Substituent position: K16 Calculated molecular weight (average): 4988.6Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1663.6, [M+4H] 4+ 1248.0 Compound No. 13 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPESGAPPPS-OH [ka] SEQ ID NO:8, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 5030.6Da LCMS_A: Retention time = 5.7 min, observed value [M+3H]3+ 1677.7, [M+4H] 4+ 1258.4 Compound No. 14 Y-Aib-EGTFTSDYSIALE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:9, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4796.4Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1599.6, [M+4H] 4+ 1199.8 Compound No. 15 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO: 10, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A, SX_A Calculated molecular weight (average): 4838.5Da LCMS_A: Retention time = 5.8 min, observed value [M+3H] 3+ 1613.4, [M+4H] 4+ 1210.3 Compound No. 16 Y-Aib-EGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(19-carboxynonadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:5, Substituent: C, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4916.5Da LCMS_A: Retention time = 6.0 min, observed value [M+3H] 3+ 1639.6, [M+4H] 4+ 1229.9 Compound No. 17 YAEGTFTSDYSIYLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:11, Substituent: B, Substituent position: K16 Synthesis method: SPPS_C, SC_A, CP_A Calculated molecular weight (average): 4874.5Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1625.4, [M+4H] 4+ 1219.4 Compound No. 18 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAG-OH [ka] SEQ ID NO:12, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4003.6Da LCMS_A: Retention time = 6.2 min, observed value [M+3H] 3+ 1335.3, [M+4H] 4+ 1001.7 Compound No. 19 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGP-OH [ka] SEQ ID NO:13, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4157.8Da LCMS_A: Retention time = 6.1 min, observed value [M+3H] 3+ 1386.6, [M+4H] 4+ 1040.3 Compound No. 20 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSS-OH [ka] SEQ ID NO:14, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4331.9Da LCMS_A: Retention time = 5.9 min, observed value [M+3H] 3+ 1444.7, [M+4H] 4+1083.7 Compound No. 21 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAR-EFINWLLAGGPSSGA-OH [ka] SEQ ID NO:15, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4460.0Da LCMS_A: Retention time = 5.9 min, observed value [M+3H] 3+ 1487.5, [M+4H] 4+ 1116.1 Compound No. 22 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFVNWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:16, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4824.4Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1608.9, [M+4H] 4+ 1206.9 Compound No. 23 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIQWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:17, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4852.5Da LCMS_A: Retention time = 5.9 min, observed value [M+3H] 3+ 1618.4, [M+4H] 4+ 1214.1 Compound No. 24 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:18, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4810.4Da LCMS_A: Retention time = 6.0 min, observed value [M+3H] 3+ 1604.0, [M+4H] 4+ 1203.4 Compound No. 25 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAEEFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:19, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4811.4Da LCMS_A: Retention time = 6.1 min, observed value [M+3H] 3+ 1604.4, [M+4H] 4+ 1203.6 Compound No. 26 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO: 20, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4819.4Da LCMS_A: Retention time = 5.8 min, observed value [M+3H] 3+ 1607.1, [M+4H] 4+ 1205.5 Compound No. 27 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPSSGAPPPS-OH [ka] SEQ ID NO:21, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4938.6Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+1646.9, [M+4H] 4+ 1235.4 Compound No. 28 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLRGGPESGAPPPS-OH [ka] SEQ ID NO:22, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A, SX_A Calculated molecular weight (average): 4980.6Da LCMS_A: Retention time = 5.8 min, observed value [M+3H] 3+ 1660.7, [M+4H] 4+ 1246.0 Compound No. 29 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAQEFIEWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:23, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A, SX_A Calculated molecular weight (average): 4825.4Da LCMS_A: Retention time = 6.1 min, observed value [M+3H] 3+ 1609.3, [M+4H] 4+ 1207.0 Compound No. 30 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAHEFIEWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:24, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4834.4Da LCMS_A: Retention time = 5.8 min, observed value [M+3H] 3+ 1612.1, [M+4H] 4+ 1209.3 Compound No. 31 Y-Aib-EGTFTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFIEWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:25, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A, SX_A Calculated molecular weight (average): 4853.5Da LCMS_A: Retention time = 5.9 min, observed value [M+3H] 3+ 1618.5, [M+4H] 4+ 1214.2 Compound No. 32 Y-Aib-EGTFTSDYSILLE-K[2-[2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFIEWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:25, Substituent: A, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4887.4Da LCMS_A: Retention time = 6.6 min, observed value [M+3H] 3+ 1629.8, [M+4H] 4+ 1222.8 Compound No. 33 Y-Aib-EGTVTSDYSILLE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:26, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4790.4Da LCMS_A: Retention time = 5.8 min, observed value [M+3H] 3+ 1597.4, [M+4H] 4+ 1198.3 Compound No. 34 Y-Aib-EGTFTSDYSIILE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:27, Substituent: B, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4838.5Da LCMS_A: Retention time = 5.8 min, observed value [M+3H] 3+ 1613.4, [M+4H] 4+ 1210.3 Compound No. 35 Y-Aib-EGTFTSDYSILLE-K[2-[2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:10, Substituent: A, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4872.4Da LCMS_A: Retention time = 6.6 min, observed value [M+3H] 3+ 1624.9, [M+4H] 4+ 1218.9 Compound No. 36 Y-Aib-EGTFTSDYSIILE-K[2-[2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-QAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:27, Substituent: A, Substituent position: K16 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4872.4Da LCMS_A: Retention time = 6.5 min, observed value [M+3H] 3+ 1624.8, [M+4H] 4+ 1218.9 Compound No. 37 Y-Aib-EGTFTSDYSIYLEEQAAR-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-FINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:28, Substituent: B, Substituent position: K21 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4888.5Da LCMS_A: Retention time = 6.0 min, observed value [M+3H] 3+ 1630.3, [M+4H] 4+ 1223.0 Compound No. 38 Y-Aib-EGTFTSDYSIYLEE-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-AAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:29, Substituent: B, Substituent position: K17 Synthesis method: SPPS_C, SC_C, CP_A Calculated molecular weight (average): 4889.5Da LCMS_A: Retention time = 6.0 min, observed value [M+3H] 3+ 1630.6, [M+4H] 4+ 1223.2 Compound No. 39 Y-Aib-EGTFTSDYSILLEEQAAR-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-FINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:30, Substituent: B, Substituent position: K21 Synthesis method: SPPS_A, SC_C, CP_A Calculated molecular weight (average): 4838.5Da LCMS_A: Retention time = 6.2 min, observed value [M+3H] 3+ 1613.5, [M+4H] 4+ 1210.4 Compound No. 40 Y-Aib-EGTFTSDYS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-LLEEQAAREFIEWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:31, Substituent: B, Substituent position: K12 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4869.4Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+ 1623.8, [M+4H] 4+ 1218.2 Compound No. 41 Y-Aib-EGTFTSDYS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-YLEEQAAREFINWLLAGGPSSGAPPPS-OH [ka] SEQ ID NO:32, Substituent: B, Substituent position: K12 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4904.4Da LCMS_A: Retention time = 5.7 min, observed value [M+3H] 3+1635.4, [M+4H] 4+ 1226.7 Compound No. 42 Y-Aib-EGTFTSDYSIYLEEQAAREFINWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH [ka] SEQ ID NO:33, Substituent: B, Substituent position: K33 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4930.5Da LCMS_A: Retention time = 6.2 min, observed value [M+3H] 3+ 1644.2, [M+4H] 4+ 1233.4 Compound No. 43 Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH [ka] SEQ ID NO:34, Substituent: B, Substituent position: K33 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4895.5Da LCMS_A: Retention time = 6.3 min, observed value [M+3H] 3+ 1632.4, [M+4H] 4+ 1224.6 Compound No. 44 Y-Aib-EGTFTSDYSILLEEQAAREFINWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH [ka] SEQ ID NO:35, Substituent: B, Substituent position: K33 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4880.5Da LCMS_A: Retention time = 6.2 min, observed value [M+3H] 3+ 1627.5, [M+4H] 4+ 1220.9 Compound No. 45 Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[2-[2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-GAPPPS-OH [ka] SEQ ID NO:34, Substituent: A, Substituent position: K33 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4929.5Da LCMS_A: Retention time = 6.6 min, observed value [M+3H] 3+ 1643.8, [M+4H] 4+ 1233.1 Compound No. 46 Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[(2S)-2-amino-6-[[(2S)-2-amino-6-[[(4S)-4-carboxy-4-(15-carboxypentadecanoylamino)butanoyl]amino]hexanoyl]amino]hexanoyl]-GAPPPS-OH [ka] SEQ ID NO:34, Substituent: E, Substituent position: K33 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4867.5Da LCMS_A: Retention time = 6.1 min, observed value [M+3H] 3+ 1623.1, [M+4H] 4+ 1217.6 Compound No. 47 Y-Aib-EGTFTSDYSILLEEQAAREFIEWLLAGGPS-K[2-[2-[2-[[2-[2-[2-[2-[[(4S)-4-carboxy-4-(15-carboxypentadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetyl]-GAPPPS-OH [ka] SEQ ID NO:34, Substituent: D, Substituent position: K33 Synthesis method: SPPS_A, SC_A, CP_A Calculated molecular weight (average): 4901.4Da LCMS_A: Retention time = 6.3 min, observed value [M+3H] 3+ 1634.6, [M+4H] 4+ 1226.1

[0152] Example 2: In vitro functional efficacy (CRE luciferase, whole cells) The purpose of this example is to test the functional activity or potency of the compound in vitro at human GLP-1 receptor and GIP receptor. In vitro functional potency is a measure of target receptor activation in whole cell assay. The potency of the derivative of Example 1 is determined as described below. Human GLP-1 (7-37) and human GIP are included in the appropriate assay for comparison.

[0153] principle In vitro functional potency was determined by measuring the response of target receptors in reporter gene assays in individual cell lines. The assay was performed in stably transfected BHK cell lines expressing one of the following G protein-coupled receptors: human GLP-1 receptor or human GIP receptor, with each cell line containing promoter-linked cAMP response element (CRE) DNA and firefly luciferase (CRE luciferase) gene. Activation of the respective receptor produces cAMP, which in turn leads to expression of luciferase protein. Once the assay incubation is complete, luciferase substrate (luciferin) is added, causing the enzymatic conversion of luciferin to oxyluciferin, resulting in bioluminescence. Luminescence is measured as the readout of the assay.

[0154] Cell culture and preparation The cell lines used in these assays were BHK cells with BHKTS13 as the parental cell line. These cells were derived from clones containing the CRE luciferase element and further established by transfecting with the respective human receptor to obtain the relevant cell line: BHK CRE luc2P hGLP-1R or BHK CRE luc2P hGIPR. Cells were cultured in cell culture medium at 37°C / 5% CO2. They were aliquoted and stored in liquid nitrogen. The cells were continuously cultured and seeded the day before each assay.

[0155] material The following chemicals were used in the assay: Pluronic F-68 10% (Gibco 2404), human serum albumin (HSA, Sigma A9511), 10% fetal bovine serum (FBS, Invitrogen 16140-071), chicken egg white ovalbumin (Sigma A5503), phenol red free DMEM (Gibco 21063-029), DMEM (Gibco 12430-054), 1M Hepes (Gibco 15630), Glutamax 100x (Gibco 35050), G418 (Invitrogen 10131-027), hygromycin (Invitrogen 10687-010), and steadylite plus (PerkinElmer 6016757).

[0156] buffer solution GLP-1R cell culture medium consisted of DMEM medium with 10% FBS, 500 μg / mL G418, and 300 μg / mL hygromycin. GIPR cell culture medium consisted of DMEM medium with 10% FBS, 400 μg / mL G418, and 300 μg / mL hygromycin. Assay buffer consisted of phenol red-free DMEM, 10 mM Hepes, 1× Glutamax, 1% ovalbumin, and 0.1% Pluronic F-68 supplemented with 2x the final assay concentration of HSA. Assay medium was mixed 1:1 with an equal volume of test compound in assay buffer to obtain the final assay concentration of HSA.

[0157] procedure 1) Cells were plated at 5000 cells / well and incubated overnight in the assay plate. 2) The cells were washed once with DPBS. 3) Test compound stocks and reference compound stocks at concentrations ranging from 100-300 μM were diluted 1:150 in assay buffer. Compounds were then diluted 1:10 in column 1 of a 96 deep-well dilution plate and then transferred to the row to generate a 3.5-fold, 12-point dilution curve. 4) Assay medium (50 μl aliquots) with or without HSA was added to each well of the assay plate. 5) 50 μl aliquots of compound or blank were transferred from the dilution plate to the assay plate containing assay buffer with or without HSA. 6) The assay plate was incubated at 37° C. in a 5% CO2 incubator for 3 hours. 7) The cells were washed once with DPBS. 8) A 100 μl aliquot of DPBS was added to each well of the assay plate. 9) A 100 μl aliquot of steadylite plus reagent (light sensitive) was added to each well of the assay plate. 10) Each assay plate was covered with aluminum foil to protect from light and shaken at 250 RPM for 30 minutes at room temperature. 11) Each assay plate was read on a microtiter plate reader.

[0158] Calculations and Results Data from the microtiter plate reader was first regression analyzed in Excel to calculate the x-axis (log scale concentration) based on the stock concentration of each test compound and the dilution in the assay. This data was then transferred to GraphPad Prism software to generate graphs and perform statistical analysis. This software performs non-linear regression (log (agonist) vs. response). EC values ​​were calculated using this software and reported in pM. 50 are shown in Tables 1-3 below. A minimum of two replicates were measured per sample. Values ​​reported are the average of the replicates. [Table 2]

[0159] The compounds of the present invention exhibit potent functional activation of the human GLP-1R receptor and the human GIPR receptor under given conditions.

[0160] Example 3: Pharmacokinetic studies in minipigs The purpose of this example is to determine the in vivo half-life of the derivatives of the invention after intravenous administration to minipigs, i.e. their time in the body and therefore their duration of action. This is done in a pharmacokinetic (PK) study to determine the terminal half-life of the derivative in question. Terminal half-life generally refers to the period of time it takes to halve a certain plasma concentration measured after the initial distribution phase.

[0161] the study Female Gottingen minipigs were obtained from Ellegaard Gottingen Minipigs (Dalmose, Denmark), were approximately 7-14 months old, and weighed approximately 16-35 kg and were used in this study. Minipigs were housed individually and fed a restricted diet of SDS minipig diet (Special Diets Services, Essex, UK) once daily.

[0162] After 3 weeks of acclimation, two permanent central venous catheters were implanted into the caval tail of each minipig. The minipig was allowed to recover for 1 week after surgery and was then used for repeated pharmacokinetic studies with appropriate washout periods between successive administrations of the derivative.

[0163] The minipigs were fasted for approximately 18 hours prior to dosing and 0-4 hours after dosing, but had free access to water during the entire period.

[0164] The sodium salt of the compound of Example 1 was dissolved in a buffer solution (pH 7.4) containing 0.007% polysorbate 20, 50 mM sodium phosphate, 70 mM sodium chloride to a concentration of 20-40 nmol / mL. An intravenous injection of the compound (usually a volume corresponding to 1.5-2 nmol / kg, e.g., 0.1 mL / kg) was given through one catheter, and blood samples were taken (preferably through the other catheter) at predetermined time points up to 14 days after administration. Blood samples (e.g., 0.8 mL) were collected in 8 mM EDTA buffer, then centrifuged at 4°C and 1942 g for 10 min.

[0165] Sampling and analysis Plasma was pipetted into Micronic tubes on dry ice and kept at -20°C until analyzed for compound plasma concentration using ELISA or similar antibody-based assay or LCMS. Individual plasma concentration-time profiles were analyzed by a non-compartmental model in Phoenix WinNonLin ver.6.4. (Pharsight Inc., Mountain View, CA, USA) and the resulting terminal half-lives (harmonic mean) were determined.

[0166] result [Table 3] The tested compounds of the present invention have very long half-lives compared to the half-lives of hGLP-1 and hGIP measured in humans, approximately 2-4 minutes and 5-7 minutes, respectively. The half-lives measured in minipigs are predictive of half-lives in humans, where at least one weekly administration by liquid injection or at least one daily administration by oral tablet is sufficient.

[0167] Example 5: Pharmacokinetic studies in dogs The purpose of this example is to determine the in vivo half-life and plasma exposure of compounds of the invention after oral administration to beagle dogs, i.e., the terminal half-life and concentration of the test substance that reaches the circulation over time. This is done in a pharmacokinetic (PK) study to determine these parameters for the compound in question. Terminal half-life generally refers to the period of time it takes to halve a particular plasma concentration measured after the initial distribution phase.

[0168] Preparation of tablet composition Tablet compositions containing the test substance and SNAC (sodium N-(8-(2-hydroxybenzoyl)amino)caprylate) were prepared according to methods known to those skilled in the art by mixing the test substance with roller compacted SNAC and magnesium stearate, for example as described in WO2019 / 149880. The amount of SNAC in the tablet composition was 100-300 mg, the amount of magnesium stearate in the tablet composition was 7.7 mg, and the target amount of each test substance in the tablet composition was 3-4 mg.

[0169] Animals, dosing, and sampling Male beagle dogs aged 1-7 years and weighing 9-17 kg during the study period were included in the study. Beagles were dosed in fasted state. Beagles were group-housed in enclosures (12-h light:12-h dark) and individually and restrictively fed Royal Canin Medium Adult dog food (Royal Canin Products, China Branch or Brogaarden A / S, Denmark) once daily. Beagles were used for repeated PK studies with suitable washout periods between successive doses. An appropriate acclimation period was allowed before the start of the first PK study. All handling, dosing, and blood sampling of beagles was performed by trained and experienced staff. Beagles were fasted overnight before the study and fasted for 0-4 hours after dosing. Beagles had limited access to water from 1 hour before dosing until 4 hours after dosing, but otherwise had free access to water throughout the entire period.

[0170] The composition was administered to groups of 6-8 beagle dogs by a single oral dose. The tablets were administered in the following manner: 10 minutes prior to tablet administration, the beagle dogs were given approximately 3 nmol / kg of SEQ ID NO: 48 subcutaneously, after which the tablet was placed at the back of the beagle's mouth to prevent chewing. The mouth was then closed and 10 mL of tap water was given via syringe or tube to facilitate swallowing of the tablet.

[0171] One blood sample was collected before dosing, and additional samples were collected at pre-defined time points after dosing, e.g., 5, 10, 15, 20, 30, and 45 min, and 1, 1.5, 2, 4, 7, 10, 24, 48, 72, 120, 144, 168, 192, 216 h, for up to 240 h to fully cover the complete plasma concentration-time absorption profile of the test substance. At each blood collection time point, approximately 0.8 mL of whole blood was collected into a 1.5 mL EDTA-coated tube, and the tube was gently swirled to mix the sample with EDTA. Blood samples were collected in EDTA buffer (8 mM) and then centrifuged at 4°C and 2000 g for 10 min. Plasma was pipetted into Micronic tubes on dry ice and kept at -20°C or lower until analysis. Blood samples were taken as needed, for example, from Venflon in the cephalic vein of the forelimb for the first 2 hours, then from the jugular vein with a syringe for the remaining time points, with the first few drops allowed to drain from the Venflon to avoid heparinized saline from the Venflon entering the sample.

[0172] All blood samples were collected in tubes containing EDTA for stabilization and kept on ice until centrifugation. Plasma was separated from whole blood by centrifugation and the plasma was stored at -20°C or below until analysis.

[0173] Analysis and Calculations Plasma was analyzed for the test substances using LC-MS (liquid chromatography-mass spectrometry) as known to those skilled in the art. The system consisted of either a Thermo Fisher QExactive mass spectrometer equipped with a 10-valve interface module TurboFlow system, a CTC HTS PAL autosampler, an Accela 1250 pump, and a Hot Pocket column oven, or a Thermo Fisher QExactive Plus mass spectrometer equipped with a valve interface module TurboFlow system, a TriPlus RSI autosampler, a Dionex UltiMate 3000 pump, and a Hot Pocket column oven. RP-HPLC separations were achieved using either a Phenomenex Onyx Monolithic C18 column (50×2.0 mm) (flow rate 0.8 mL / min at 30° C.) or an Agilent Poroshell 120 SB-C18 column (50×2.1 mm, 2.7 μm) (flow rate 0.4 mL / min at 60° C.) using a linear gradient of 1:1 acetonitrile / methanol in 1% aqueous formic acid. The mass spectrometer was operated in either positive ionization SIM mode or positive ionization PRM mode.

[0174] For each individual beagle dog, the plasma concentration-time profile was analyzed by a noncompartmental model in Pharsight Phoenix WinNonLin version 6.4 software or other relevant software for PK analysis, and the resulting terminal half-lives (t 1 / 2 ), maximum plasma concentration per dose (C max / D), time to maximum plasma concentration (t max ), and area under the curve to maximum per dose (AUC / D) were determined. Summary statistics of the pharmacokinetic results were reported as median (t max ), hormonal mean (t 1 / 2 ), or the arithmetic mean (C max , AUC).

[0175] result [Table 4] Concentrations of the compound were detected in plasma after oral administration (C max The tested compounds of the invention show oral bioavailability in this model, since AUC / D=0 and AUC / D=0. Moreover, the tested compounds of the invention further have very long half-lives compared to the half-lives of hGLP-1 and hGIP measured in humans, which are approximately 2-4 minutes and 5-7 minutes, respectively.

[0176] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those skilled in the art. It is therefore to be understood that the appended embodiments are intended to cover all such modifications and changes that fall within the true spirit of the invention.

Claims

1. A compound comprising a peptide and a substituent, the amino acid sequence of the peptide being: YX ２ EGTX ６ TSDYSX １２ X １３ LEX １６ QAAX ２０ X ２１ FX ２３ X ２４ WLLX ２８ GX ３０ X ３１ X ３２ X ３３ X ３４ X ３５ X ３６ X ３７ X ３８ X ３９ (SEQ ID NO: 47) with an optional amide modification at the C-terminus, In the formula, X ２ is Aib or A, X ６ is F or V, X １２ is I or Y, X １３ is Y, A, L, or I; X １６ is K or E, X ２０ are Q, R, E, and H, X ２１ is A or E, X ２３ is I or V, X ２４ is E, Q, or N; X ２８ is A or R, X ３０ is G or absent, X ３１ is P or absent, X ３２ is E, S, or absent; X ３３ is S, K, or absent, X ３４ is G or absent, X ３５ is A or absent, X ３６ is P or absent, X ３７ is P or absent, X ３８ is P or absent, X ３９ is S or absent, the substituent is attached to the peptide via a lysine (K) at position 16 or 33, or a pharma- ceutically acceptable salt thereof.

2. X ３２ X ３３ X ３４ X ３５ The compound of claim 1 , wherein is selected from the group of SSGA, ESGA, and SKGA.

3. The amino acid sequence of the peptide is YX ２ EGT ６ TSDYSX １２ X １３ LEX １６ QAAX ２０ X ２１ Forex ２３ X ２４ WLLX ２８ GGPSSGAPPPS (SEQ ID NO:37) During the ceremony, X ２ is Aib or A, X ６ is F or V, X １２ is I or Y, X １３ is Y, A, L, or I; X １６ is K or E, X ２０ are Q, R, E, and H, X ２１ is A or E, X ２３ is I or V, X ２４ is E, Q, or N; X ２８ is A or R.

4. X １３ LEX １６ QAAX ２０ X ２１ Forex ２３ X ２４ The compound of any one of claims 1 to 3, wherein is selected from the group consisting of LLEKQAAREFIN, LLEKQAAREFIE, LLEKQAAQEFIE, and LLEEQAAREFIE.

5. The compound of claim 1, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35.

6. The compound of claim 5 , wherein the peptide has the amide modification at the C-terminus.

7. The compound according to any one of claims 1 to 6, wherein the substituents are selected from the group consisting of: 【Chemistry 1】

8. 2. The compound of claim 1, wherein the compound is selected from the group consisting of: 【Chemistry 2-1】 【Chemistry 2-2】 【Chemistry 2-3】 【Chemistry 2-4】 【Chemistry 2-5】 【Chemistry 2-6】 【Chemistry 2-7】 【Chemistry 2-8】 【Chemistry 2-9】

9. 2. The compound of claim 1, wherein the compound is selected from the group consisting of: 【Table 1-1】 【Table 1-2】

10. 2. The compound of claim 1, wherein the compound is: 【Table 2】

11. A compound according to any one of claims 1 to 10 for use as a medicament.

12. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10 for the prevention and / or treatment of diabetes and / or obesity.

13. 11. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10 for the prevention and / or treatment of liver disorders such as hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH) liver inflammation and / or fatty liver.

14. A peptide having the following amino acid sequence: YX ２ EGTX ６ TSDYSX １２ X １３ LEX １６ QAAX ２０ X ２１ FX ２３ X ２４ WLLX ２８ GX ３０ X ３１ X ３２ X ３３ X ３４ X ３５ X ３６ X ３７ X ３８ X ３９ (SEQ ID NO: 47) with an optional amide modification at the C-terminus, In the formula, X ２ is Aib or A, X ６ is F or V, X １２ is I or Y, X １３ is Y, A, L, or I; X １６ is K or E, X ２０ are Q, R, E, and H, X ２１ is A or E, X ２３ is I or V, X ２４ is E, Q, or N; X ２８ is A or R, X ３０ is G or absent, X ３１ is P or absent, X ３２ is E, S, or absent; X ３３ is S, K, or absent, X ３４ is G or absent, X ３５ is A or absent, X ３６ is P or absent, X ３７ is P or absent, X ３８ is P or absent, X ３９ is S or absent.

15. The peptide of claim 14, wherein the amino acid sequence of the peptide is any one of SEQ ID NOs: 1-27 or 33-35.

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