GLP-1 / GIP dual, GLP-1 / GCG dual, and GLP-1 / GIP / GCG triple receptor agonists

Abstract

The present invention relates to GLP-1 / GIP / GCG triple receptor agonists and their use in the treatment or prevention of type 2 diabetes (T2DM), hyperlipidemia / dyslipidemia, metabolic syndrome, metabolic dysfunction-related fatty liver disease (MASLD), metabolic dysfunction-related steatohepatitis (MASH), neurodegenerative disorders, fibrosis, cardiovascular risk and / or obesity.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to Indian application No. 202321039646, filed on June 9, 2023, which is incorporated herein by reference in its entirety.

[0003] Reference to the electronic sequence list

[0004] This application includes a sequence list submitted electronically, which is incorporated herein by reference in its entirety. The sequence list was created on June 7, 2024, named "24-0750-WO_Sequence-Listing.xml", and is 143,360 bytes in size. Technical Field

[0005] This disclosure relates to GLP-1 / GLP dual, GLP-1 / GCG dual, and GLP-1 / GIP / GCG triple receptor agonists. In particular, this disclosure relates to GLP-1 / GIP / GCG triple receptor agonists comprising an incretin analog peptide. The peptides described herein have structural features that provide balanced activity and prolonged duration of action on each of these receptors. The peptides according to this disclosure can be used to treat type 2 diabetes mellitus (T2DM), hyperlipidemia / dyslipidemia, metabolic syndrome, metabolic dysfunction-associated fatty liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), neurodegenerative diseases, fibrosis, obesity, and to reduce cardiovascular risk. Background Technology

[0006] Over the past few decades, the prevalence of diabetes has been steadily increasing. Type 2 diabetes mellitus (T2DM) is the most common form of diabetes, characterized by high blood sugar levels caused by insulin resistance. People with T2DM are more likely to develop comorbidities such as hyperlipidemia / dyslipidemia, metabolic syndrome, metabolic dysfunction-associated fatty liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), neurodegenerative diseases, fibrosis, cardiovascular risk, and / or obesity.

[0007] Current treatments for type 2 diabetes mellitus (T2DM) include diet and exercise, as well as oral and injectable medications to lower blood sugar, including insulin-based therapies such as GLP-1 monoreceptor agonists and / or GLP-1 / GIP dual-receptor agonists. As an emerging approach, new therapies are being investigated in which compounds are active not only against GLP-1 monoreceptors or GLP-1 / GIP dual-receptors, but also against GCG receptors. Some compounds have been described as having dual GLP-1 / GCG and / or triple GLP-1 / GIP / GCG agonist activity.

[0008] For example, cotadutide, MK-1462, and mazdutide are peptides that act as dual GLP-1 / GCG receptor agonists. Similarly, retatrutide (SEQ ID NO: 7) is another clinical candidate that acts as a triple GLP-1 / GIP / GCG receptor agonist. WIPO publications WO2019 / 193576, WO2006 / 097537, and WO1998 / 008871 disclose GLP-1 receptor agonist compounds. WIPO publications WO2022 / 079639, WO2021 / 260530, WO2017 / 74714A1, WO2020 / 23386, WO2020 / 023388, WO2015 / 067715, WO2016 / 111971, WO2014 / 192284, WO2011 / 119657, and WO2013 / 164483 disclose GLP-1 / GIP dual receptor agonist compounds. WIPO publications WO2011 / 075393, WO2012 / 177444, WO2014 / 091316, and WO2017 / 153575 disclose GLP-1 / GCG dual receptor agonist compounds. WIPO publications WO2015 / 067716, WO2016 / 198624, WO2014 / 049610 and WO2017 / 116204 disclose GLP-1 / GIP / GCG triple receptor agonist compounds.

[0009] Recent studies on GLP-1 / GCG dual and / or GLP-1 / GIP / GCG triple receptor agonists have also highlighted the importance of understanding the contribution and different effects of individual hormones by altering GLP-1:GCG activity and the ratio in GLP-1 / GCG dual and GLP-1 / GIP / GCG triple receptor agonists. Hope et al. Front. Endocrinol. 08 September 2021, Vol 12-2021 .

[0010] It is known that glucagon receptor (GCGR) agonism, in addition to causing diabetes, increases heart rate and contractility, which may lead to adverse cardiovascular outcomes. Further chronic excess of glucagon also leads to the catabolism of amino acids and proteins, resulting in the loss of lean body mass.

[0011] For example, while retaliglutide has shown increased weight loss in the treatment of obesity, it is associated with several side effects such as increased heart rate. These adverse effects may be attributed to the GCGR agonist component of the drug's action. Current research in Diabetes & Obesity Journal, July 26, 2023 .

[0012] Therefore, while the broad metabolic benefits of GLP-1, GIP, and GCG receptor agonist compounds have been established in therapeutic paradigms, there remains a need for treatments that can provide effective glucose control with weight loss benefits and a reduced profile of adverse effects such as increased heart rate, arrhythmias, and loss of lean body mass, particularly for the treatment of type 2 diabetes mellitus (T2DM) and related comorbidities such as cardiovascular disease and / or obesity. There is also a need for therapeutic agents that can be used with a sufficiently prolonged duration of action to allow for infrequent dosing, either twice weekly or once weekly.

[0013] It has been surprisingly found that modifications to fatty acid side chains and / or substitution of branched side-chain amino acids with straight-chain side-chain amino acids in the sequences of GLP-1 / GCG dual and / or GLP-1 / GIP / GCG triple receptor agonists can produce significant GLP-1 advantage, which enables enhanced weight loss without a sharp reduction in food intake and reduced side effects. Summary of the Invention

[0014] In one aspect, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 1) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is the L or D isomer of the amino acid of the following formula: ,in" “ indicates the connection point with Leu, and R is selected from C1-C6 alkyl, C3-C6 cycloalkylmethyl and C3-C6 cycloalkyl; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is the L or D isomer of K, Aib, or an amino acid of the following formula: ,in" “ indicates the connection point with Leu, and R is selected from C1-C6 alkyl, C3-C6 cycloalkylmethyl and C3-C6 cycloalkyl; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated at the C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and at least one of K is related to C. 16 -C 22 Fatty acid conjugation.

[0015] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 2) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is αMe-L; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is the L or D isomer of K, Aib, or an amino acid of the following formula: ,in" “ indicates the connection point with Leu, and R is selected from C1-C6 alkyl, C3-C6 cycloalkylmethyl and C3-C6 cycloalkyl; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; X28 is either A or E; X29 is G X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and the further condition is that at least one of K contains a partially acylated side chain amino group selected from the following formula ( amino group: Aminoethoxyethoxyacetic acid-Aib-Glu-C 16 -C 22 fatty acid chains; Aminoethoxyethoxyacetic acid-C(O)-diaminobutane-Glu-C 16 -C 22 fatty acid chains; Glu-C 16 -C 22 fatty acid chains; Aminoethoxyethoxyacetic acid-aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 fatty acid chains; and Aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 Fatty acid chains.

[0016] The condition is that when X20 is Aib, the side-chain amino group is not affected by aminoethoxyethoxyacetic acid-aminoethoxyethoxyacetic acid-Glu-C. 16 -C 22 Fatty acid chain or aminoethoxyethoxyacetic acid-Glu-C16 -C 22 Fatty acid chain acylation.

[0017] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 3) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is Aib, Ser(OMe), nor-V, nor-L, or αMe-L; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is K, Aib, Ser(OMe), nor-V, or nor-L; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is E, I, or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and the further condition is that at least one of K contains a partially acylated side chain amino group selected from the following formula ( amino group:

[0018] The polypeptide described herein is not SEQ ID NO: 7, SEQ ID NO: 23 or SEQ ID NO: 30.

[0019] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 4) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is nor-V, nor-L, or αMe-L; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is either K or Aib; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; Each of X36, X37, and X38 is independently P; and X39 is S; The condition is that at least one of X17 and X20 is K, and the further condition is that said K contains a partially acylated side chain amino group selected from the following formula ( amino group:

[0020] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: Y-Aib-X3-GTFTSDYSI-X13-LDK-X17-AQ-X20-AF-X23-X24-X25-L-X27-X28-GGPSSGAPPPS (SEQ ID NO: 5) in: X3 is either Q or N; X13 is nor-V, nor-L, or αMe-L; X17 is either I or K; X20 is either K or Aib; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; and X28 is either A or E; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and the further condition is that said K contains a partially acylated side chain amino group selected from the following formula ( amino group:

[0021] The polypeptide mentioned therein is not SEQ ID NO: 7.

[0022] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: Y-Aib-X3-GTFTSDYSI-X13-LDK-X17-AQ-X20-AFIEYLLEGGPSSGAPPPS (SEQ ID NO: 6) in: X3 is either Q or N; X13 is nor-V, nor-L, or αMe-L; X17 is K; and X20 is Aib, nor-L, or nor-V; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0023] The polypeptide mentioned therein is not SEQ ID NO: 7.

[0024] On the other hand, this disclosure relates to an incretin analog polypeptide comprising: Lysine residues containing fatty acid elongation groups attached to the ε-nitrogen of lysine; A peptide residue containing the sequence Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31), which is indirectly linked to the lysine residue via its carboxyl terminus; A peptide residue having the sequence Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32), which is indirectly linked to the carboxyl group of the lysine residue via the amino terminus of the Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue; and A valine amino acid residue that is indirectly linked to and located between the Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) residue and the lysine residue.

[0025] On the other hand, this disclosure relates to an incretin analogue comprising: Lysine residues containing a group of formula (I) linked to the ε-nitrogen of lysine. Where equation (I) is (I) in: U does not exist or represents -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the connection point with W; W indicates -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], -C(O)-NH-(CH2) 3-4 -NH-], -C(O)-C(CH3)2-NH-] or , Where ] is the connection point with Y; Y does not exist or represents -C(O)-(CH2)2-CH(CO2H)NH-- or -C(O)CH((CH2) x CO2H)NH--, where x is 1, 2, or 3, and -- is the connection point with Z; and Z represents -C(O)-(CH2) n -COOH or -C(O)-(CH2) n -CH3, where n is an integer from 14 to 20; A peptide residue containing the sequence Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31), which is indirectly linked to the lysine residue via its carboxyl terminus; A peptide residue having the sequence Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2 (SEQ ID NO: 32), which is indirectly linked to the carboxyl group of the lysine residue via the amino terminus of the Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue; and A valine amino acid residue that is indirectly linked to and located between a Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 32) residue and the lysine residue. Detailed Implementation

[0026] abbreviations

[0027] Aib: 2-Aminoisobutyric acid

[0028] DIPEA: N , N '-Diisopropylethylamine

[0029] HOBt: 1-Hydroxybenzotriazole

[0030] DIPC: N , N '-Diisopropylcarbodiimide

[0031] THF: Tetrahydrofuran

[0032] DCM: Dichloromethane

[0033] Fmoc: fluorenylmethoxycarbonyl

[0034] HOSu: N- Hydroxysuccinimide

[0035] DCC: Dicyclohexylcarbodiimide

[0036] DMAc: dimethylacetamide

[0037] IBCF: Isobutyl chloroformate

[0038] NMM: N -Methylmorpholine

[0039] DIC: Diisopropylcarbodiimide

[0040] definition

[0041] The term "pharmaceutically acceptable salt" according to this disclosure includes acid addition salts formed with organic or inorganic acids. Suitable pharmaceutically acceptable salts of the compounds of this disclosure include acid addition salts, which may be salts of inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, etc., or salts of organic acids such as acetic acid, benzenesulfonic acid, methanesulfonic acid, benzoic acid, citric acid, lactic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, octanoic acid, azelaic acid, malic acid, tartaric acid, amino acids (e.g., glutamic acid or aspartic acid), etc. Pharmaceutically acceptable acid addition salts of the compounds of this disclosure include salts formed by adding one or more equivalent amounts of acid, such as monohydrochloride or dihydrochloride. Salts can be prepared by any method within the knowledge of those skilled in the art. See Berge et al. , J. Pharm. Sci. , 1977, 66, 1-19; and "Handbook ofPharmaceutical Salts: Properties, Selection, and Use," Edited by Stahl et al. , Verlag Helv. Chim. Acta Zurich, Switzerland, and Wiley-VCH (Weinheim, Germany, 2002.)

[0042] As used in this article, terminology " "Effective dose" or "effective... dose" refers to an amount of compound that, when administered to a subject in a single or multiple doses, is sufficient to cure, alleviate, relieve, or partially resolve the clinical manifestations of a given disease or symptom and its complications, exceeding the clinical manifestations expected in the absence of such treatment. Therefore, the result may be a reduction and / or relief of signs, symptoms, or causes of the disease, or any other desired biological alteration. It should be understood that "therapeutic effective dose" can vary from subject to subject, depending on the subject's age, weight, general condition, the condition being treated, the severity of the condition being treated, and the prescribing physician's judgment.

[0043] As used in this article, the amino acid "Aib" can be represented by the following structure: It can also be defined by its chemical name, "2-aminoisobutyric acid".

[0044] As used in this article, the amino acid “S(OMe)” or “Ser(OMe)” can be represented by the following structure: It can also be defined by its chemical name, "serine methyl ether". The terms L-Ser(OMe) and D-Ser(OMe) refer to the "L" and "D" isomers of Ser(OMe), respectively.

[0045] As used herein, the amino acid “nor-V”, “nor-Val”, or “norvaline” can be represented by the following structure: It can also be defined by its chemical name, "2-aminovaleric acid". The terms L-valine and D-valine refer to the "L" and "D" isomers of valine, respectively.

[0046] As used in this article, the amino acid “nor-L”, “nor-Leu”, or “ortholeucine” can be represented by the following structure: It can also be defined by its chemical name, "2-aminohexanoic acid". The terms L-leucine and D-leucine refer to the "L" and "D" isomers of leucine, respectively.

[0047] As used in this article, the amino acid “αMe-L”, “αMe-Leu”, or “αMe-Leucine” can be represented by the following structures: It can also be defined by its chemical name, "2-amino-2,4-dimethylvaleric acid". The terms L-α-Me-leucine and D-α-Me-leucine refer to the "L" and "D" isomers of α-Me-leucine, respectively.

[0048] As described herein, this disclosure provides stable, long-acting GLP-1 monoreceptor, GLP-1 / GIP dual-receptor, GLP-1 / GCG dual-receptor and / or GLP-1 / GIP / GCG triple-receptor agonists that can be used to treat type 2 diabetes mellitus (T2DM), hyperlipidemia / dyslipidemia, metabolic syndrome, metabolic dysfunction-associated fatty liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), neurodegenerative diseases, fibrosis and / or obesity, as well as to reduce cardiovascular risk.

[0049] In one aspect, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 1) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is the L or D isomer of the amino acid of the following formula: ,in" “ indicates the connection point with Leu, and R is selected from C1-C6 alkyl, C3-C6 cycloalkylmethyl and C3-C6 cycloalkyl; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is the L or D isomer of K, Aib, or an amino acid of the following formula: ,in" “ indicates the connection point with Leu, and R is selected from C1-C6 alkyl, C3-C6 cycloalkylmethyl and C3-C6 cycloalkyl; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and at least one of K is related to C. 16 -C 22 Fatty acid conjugation.

[0050] In one embodiment, the polypeptide of SEQ ID NO: 1 may have a linker with C 16 -C 22 K conjugated with fatty acids.

[0051] In some embodiments, the connector is selected from the group consisting of aminoethoxyethoxyacetic acid, glutamic acid, diaminobutane, Aib, and any combination thereof.

[0052] In a preferred embodiment, the glutamic acid is γ-glutamic acid.

[0053] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 2) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is αMe-L; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is the L or D isomer of K, Aib, or an amino acid of the following formula: ,in" “ indicates the connection point with Leu, and R is selected from C1-C6 alkyl, C3-C6 cycloalkylmethyl and C3-C6 cycloalkyl; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and the further condition is that at least one of K contains a partially acylated side chain amino group selected from the following formula ( amino group: Aminoethoxyethoxyacetic acid-Aib-Glu-C 16 -C 22 fatty acid chains; Aminoethoxyethoxyacetic acid-C(O)-diaminobutane-Glu-C 16 -C 22 fatty acid chains; Glu-C 16 -C 22 fatty acid chains; Aminoethoxyethoxyacetic acid-aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 fatty acid chains; and Aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 Fatty acid chains.

[0054] The condition is that when X20 is Aib, the side-chain amino group is not affected by aminoethoxyethoxyacetic acid-aminoethoxyethoxyacetic acid-Glu-C. 16 -C 22 Fatty acid chain or aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 Fatty acid chain acylation.

[0055] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 3) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is Aib, Ser(OMe), nor-V, nor-L, or αMe-L; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is K, Aib, Ser(OMe), nor-V, or nor-L; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is E, I, or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and the further condition is that at least one of K has a partially acylated side chain amino group selected from the following formula ( amino group:

[0056] The polypeptide described herein is not SEQ ID NO: 7, SEQ ID NO: 23 or SEQ ID NO: 30.

[0057] In one embodiment, the polypeptide according to SEQ ID NO: 3 may comprise: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is nor-V, nor-L, or αMe-L; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is K, Aib, nor-V, or nor-L; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; Each of X36, X37, and X38 is independently P; and X39 is S; The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0058] The polypeptide mentioned therein is not SEQ ID NO: 7.

[0059] In another embodiment, the polypeptide according to SEQ ID NO: 3 may comprise: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is nor-V, nor-L, or αMe-L; X16 is K; X17 is K; X18 is A; X19 is Q; X20 is Aib, nor-V, or nor-L; X21 is A; X23 is I; X24 is E; X25 is Y; X27 is L; X28 is E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; Each of X36, X37, and X38 is independently P; and X39 is S; The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0060] The polypeptide mentioned therein is not SEQ ID NO: 7.

[0061] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: X1-X2-X3-GTFTSD-X10-S-X12-X13-LD-X16-X17-X18-X19-X20-X21-F-X23-X 24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 4) in: X1 is Y; X2 is Aib; X3 is either Q or N; X10 is Y; X12 is I; X13 is nor-V, nor-L, or αMe-L; X16 is K; X17 is either I or K; X18 is A; X19 is Q; X20 is either K or Aib; X21 is A; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; X28 is either A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; Each of X36, X37, and X38 is independently P; and X39 is S; The condition is that at least one of X17 and X20 is K, and the further condition is that said K contains a partially acylated side chain amino group selected from the following formula ( amino group:

[0062] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: Y-Aib-X3-GTFTSDYSI-X13-LDK-X17-AQ-X20-AF-X23-X24-X25-L-X27-X28-GGPSSGAPPPS (SEQ ID NO: 5) in: X3 is either Q or N; X13 is nor-V, nor-L, or αMe-L; X17 is either I or K; X20 is either K or Aib; X23 is either V or I; X24 is either Q or E; X25 is either W or Y; X27 is either I or L; and X28 is either A or E; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The condition is that at least one of X17 and X20 is K, and the further condition is that said K contains a partially acylated side chain amino group selected from the following formula ( amino group:

[0063] The polypeptide mentioned therein is not SEQ ID NO: 7.

[0064] In one embodiment, the polypeptide according to SEQ ID NO: 5 may comprise: X3 is Q; X13 is αMe-L; X17 is K; X20 is AiB; X23 is I; X24 is E; X25 is Y; X27 is L; and X28 is E; The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0065] In another embodiment, the polypeptide according to SEQ ID NO: 5 may comprise: X3 is Q; X13 is NOR-V; X17 is K; X20 is AiB; X23 is I; X24 is E; X25 is Y; X27 is L; and X28 is E; The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0066] In another embodiment, the polypeptide according to SEQ ID NO: 5 may comprise: X3 is Q; X13 is NOR-L; X17 is K; X20 is AiB; X23 is I; X24 is E; X25 is Y; X27 is L; and X28 is E; The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0067] In another embodiment, the polypeptide according to SEQ ID NO: 5 may comprise: X3 is Q; X13 is NOR-V; X17 is I; X20 is K; X23 is V; X24 is Q; X25 is W; X27 is I; and X28 is A; The side chain amino group of K at position X20 ( The amino group is partially acylated using the following formula:

[0068] In another embodiment, the polypeptide according to SEQ ID NO: 5 may comprise: X3 is N; X13 is NOR-V; X17 is I; X20 is K; X23 is V; X24 is Q; X25 is W; X27 is I; and X28 is A; The side chain amino group of K at position X20 ( The amino group is partially acylated using the following formula:

[0069] On the other hand, this disclosure relates to a polypeptide comprising the following amino acid sequence or a pharmaceutically acceptable salt thereof: Y-Aib-X3-GTFTSDYSI-X13-LDK-X17-AQ-X20-AFIEYLLEGGPSSGAPPPS (SEQ ID NO: 6) in: X3 is either Q or N; X13 is nor-V, nor-L, or αMe-L; X17 is K; and X20 is Aib, nor-L, or nor-V; The acid group of the C-terminal amino acid is either a free carboxylic acid group or is amidated to a C-terminal primary amide; and The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0070] The polypeptide mentioned therein is not SEQ ID NO: 7.

[0071] In one embodiment, the polypeptide according to SEQ ID NO: 6 may comprise: X13 is either nor-V or nor-L; and X20 is AiB; The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0072] In another embodiment, the polypeptide according to SEQ ID NO: 6 may comprise: X13 is αMe-L; and X20 is either nor-L or nor-V; The side chain amino group of K at position X17 ( The amino group is partially acylated using the following formula:

[0073] On the other hand, this disclosure relates to an incretin analog polypeptide comprising: Lysine residues containing fatty acid elongation groups attached to the ε-nitrogen of lysine; A peptide residue containing the sequence Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31), which is indirectly linked to the lysine residue via its carboxyl terminus; A peptide residue having the sequence Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32), which is indirectly linked to the carboxyl group of the lysine residue via the amino terminus of the Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue; and A valine amino acid residue that is indirectly linked to and located between the Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) residue and the lysine residue.

[0074] On the other hand, this disclosure relates to an incretin analogue comprising: A lysine residue comprising a group of formula (I) linked to the ε-nitrogen of lysine, wherein formula (I) is

[0075] (I)

[0076] in: U does not exist or represents -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, where} is the connection point with W; W indicates -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], -C(O)-NH-(CH2) 3-4 -NH-], -C(O)-C(CH3)2-NH-] or , Where ] is the connection point with Y; Y does not exist or represents -C(O)-(CH2)2-CH(CO2H)NH-- or -C(O)CH((CH2) x CO2H)NH--, where x is 1, 2, or 3, and -- is the connection point with Z; and Z represents -C(O)-(CH2) n -COOH or -C(O)-(CH2) n -CH3, where n is an integer from 14 to 20; A peptide residue containing the sequence Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31), which is indirectly linked to the lysine residue via its carboxyl terminus; A peptide residue having the sequence Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2 (SEQ ID NO: 32), which is indirectly linked to the carboxyl group of the lysine residue via the amino terminus of the Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue; and A valine amino acid residue that is indirectly linked to and located between the Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) residue and the lysine residue.

[0077] In one embodiment, in the incretin analogue, lysine is linked to a Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) residue via a peptide residue comprising 10 amino acids.

[0078] In another embodiment, in the incretin analogue, lysine is linked to the Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue via a peptide residue comprising 11 amino acids.

[0079] On the other hand, this disclosure relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide or incretin analog as described herein.

[0080] In another aspect, this disclosure relates to a method for treating obesity, type 2 diabetes mellitus (T2DM), metabolic syndrome, metabolic dysfunction-associated fatty liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), neurodegenerative diseases, fibrosis, hyperlipidemia / dyslipidemia, obesity, and reducing cardiovascular risk, said method comprising administering a polypeptide or incretin analogue as described herein to a patient in need of such treatment.

[0081] On the other hand, this disclosure relates to a polypeptide comprising an amino acid sequence selected from the group consisting of: i.) Y-Aib-QGTFTSDYSI-(αMe-L)-LDKK AQ-Aib-AFIEYLLEGPSSGAPPPS-NH2 (SEQ ID NO: 8); ii.) Y-Aib-QGTFTSDYSI-(valine)-LDKK AQ-Aib-AFIEYLLEGPSSGAPPPS-NH2 (SEQ ID NO: 9); iii.) Y-Aib-NGTFTSDYSI-(valine)-LDKK AQ-Aib-AFIEYLLEGPSSGAPPPS-NH2 (SEQ ID NO: 10); iv.) Y-Aib-QGTFTSDYSI-(ortholeucine)-LDKK AQ-Aib-AFIEYLLEGPSSGAPPPS-NH2 (SEQ ID NO: 11); v.) Y-Aib-QGTFTSDYSI-(αMe-L)-LDKK AQ-(nor-V)-AFIEYLLEGPGPSSGAPPPS-NH2 (SEQ ID NO: 12); vi.) Y-Aib-NGTFTSDYSI-(αMe-L)-LDKK AQ-(nor-V)-AFIEYLLEGPSSGAPPPS-NH2 (SEQ ID NO: 13); vii.) Y-Aib-QGTFTSDYSI-(αMe-L)-LDKK AQ-(nor-L)-AFIEYLLEGPGPSSGAPPPS-NH2 (SEQ ID NO: 14); viii.) Y-Aib-QGTFTSDYSI-(N-valine)-LDKIAQK AFVQWLIAGGPSSGAPPPS-NH2 (SEQ ID NO: 15); and ix.) Y-Aib-NGTFTSDYSI-(norvaline)-LDKIAQK AFVQWLIAGGPSSGAPPPS-NH2 (SEQID NO: 16), Where K The side chain amino group ( The amino group is partially acylated using the following formula:

[0082] The polypeptide described herein is not SEQ ID NO: 7, SEQ ID NO: 23 or SEQ ID NO: 30.

[0083] The sequences of the polypeptides described herein are represented by single-letter or three-letter codes of amino acids approved by the International Union of Pure and Applied Chemistry (IUPAC).

[0084] Unless otherwise stated, this disclosure is intended to cover L and D isomers of the amino acids in the sequences described herein. However, in some preferred embodiments, all amino acids are in the “L” configuration unless otherwise stated.

[0085] In another respect, this disclosure relates to a polypeptide selected from one of the representative compounds in Table 1 or a pharmaceutically acceptable salt thereof.

[0086] Table 1. Representative polypeptide compounds Unless otherwise stated, all amino acids mentioned in Table 1 above are in the “L” configuration.

[0087] Table 2. Structure of Parts A, B, C, D, E, F, G, H, and I

[0088] On the other hand, this disclosure relates to a method for treating or preventing type 2 diabetes mellitus (T2DM).

[0089] On the other hand, this disclosure relates to a method for treating or preventing hyperlipidemia / dyslipidemia.

[0090] On the other hand, this disclosure relates to a method for treating or preventing obesity.

[0091] In another aspect, this disclosure relates to a method for treating or preventing metabolic syndrome, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), neurodegenerative diseases, fibrosis, and / or cardiovascular risks.

[0092] In one implementation, the treatment method includes administering an effective amount of the polypeptide or a pharmaceutically acceptable salt thereof, as described herein, to a patient in need.

[0093] In another aspect, this disclosure relates to a method for treating type 2 diabetes mellitus (T2DM), the method comprising administering to a patient in need of such treatment an effective amount of a polypeptide as described herein or a pharmaceutically acceptable salt thereof.

[0094] In another respect, this disclosure relates to a method for treating obesity, the method comprising administering to a patient in need of such treatment an effective amount of a polypeptide or a pharmaceutically acceptable salt thereof as described herein.

[0095] In another aspect, this disclosure relates to a method for treating hyperlipidemia / dyslipidemia, the method comprising administering to a patient in need of such treatment an effective amount of a polypeptide or a pharmaceutically acceptable salt thereof as described herein.

[0096] In another aspect, this disclosure relates to a pharmaceutical composition comprising one or more of a polypeptide or a pharmaceutically acceptable salt thereof as described herein, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0097] The compounds of the present invention are preferably formulated into pharmaceutical compositions for administration via parenteral routes (e.g., subcutaneous, intravenous, intraperitoneal, intramuscular, or transdermal). Such pharmaceutical compositions and methods of their preparation are well known in the art. See, for example, "Remington: The Science and 50 Practice of Pharmacy, " DB Troy Edited, 21st Edition Lippincott, Williams & Wilkins , 2006).

[0098] On the other hand, this disclosure relates to polypeptides as described herein or pharmaceutically acceptable salts thereof used as medicines.

[0099] In another respect, this disclosure relates to polypeptides as described herein or pharmaceutically acceptable salts thereof used for the treatment or prevention of type 2 diabetes mellitus (T2DM).

[0100] On the other hand, this disclosure relates to polypeptides as described herein or pharmaceutically acceptable salts thereof used for the treatment or prevention of hyperlipidemia / dyslipidemia.

[0101] On the other hand, this disclosure relates to polypeptides or pharmaceutically acceptable salts thereof, as described herein, for use in the treatment or prevention of obesity.

[0102] On the other hand, this disclosure relates to the use of polypeptides or pharmaceutically acceptable salts thereof described herein for the treatment or prevention of diseases selected from the group consisting of: metabolic syndrome, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), neurodegenerative diseases, fibrosis, and cardiovascular risks.

[0103] On the other hand, the polypeptides or pharmaceutically acceptable salts thereof described herein may be administered simultaneously, separately or sequentially in combination with effective amounts of one or more other therapeutic agents.

[0104] On the other hand, the pharmaceutical composition according to this disclosure comprises a polypeptide or a pharmaceutically acceptable salt thereof as described herein, used as a medicament.

[0105] On the other hand, the pharmaceutical compositions according to this disclosure comprise a polypeptide or a pharmaceutically acceptable salt thereof as described herein for the treatment or prevention of type 2 diabetes mellitus (T2DM).

[0106] On the other hand, the pharmaceutical compositions according to this disclosure comprise a polypeptide or a pharmaceutically acceptable salt thereof as described herein, used for the treatment or prevention of hyperlipidemia / dyslipidemia.

[0107] On the other hand, the pharmaceutical compositions according to this disclosure comprise a polypeptide or a pharmaceutically acceptable salt thereof, as described herein, used for the treatment or prevention of obesity.

[0108] On the other hand, the pharmaceutical compositions according to this disclosure comprise a polypeptide or a pharmaceutically acceptable salt thereof as described herein for the treatment or prevention of diseases selected from the group consisting of: metabolic syndrome, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), neurodegenerative diseases, fibrosis, and cardiovascular risks.

[0109] On the other hand, the pharmaceutical composition according to this disclosure comprises a polypeptide or a pharmaceutically acceptable salt thereof, as described herein, provided simultaneously, separately or sequentially with an effective amount of one or more other therapeutic agents.

[0110] This disclosure may relate to one or more embodiments as described herein. It should be understood that the embodiments described herein are illustrative of this disclosure and are not intended to limit the claims to the specific embodiments exemplified. It should also be understood that the embodiments as defined herein may be used independently or in combination with any definition or any other embodiment as defined herein. Therefore, this disclosure contemplates all possible combinations and permutations of the various independently described embodiments.

[0111] Other features of this disclosure will become apparent to those skilled in the art based on the following embodiments. In general, this disclosure can be extended to any novel features as described herein, including the appended claims and drawings. Therefore, features, integers, properties, compounds, or chemical portions described in conjunction with a particular aspect, embodiment, or example of this disclosure should be understood to be applicable to any other aspect, embodiment, or example as described herein, unless incompatible therewith.

[0112] Furthermore, unless otherwise stated, any feature disclosed herein may be replaced by an alternative feature serving the same or similar purpose.

[0113] Example

[0114] Instruments and analytical methods. The instruments used for characterizing and analyzing the compounds described herein include a high-performance liquid chromatograph (HPLC) (Waters e2695 Alliance; detector: Waters (2489 UV / visible)).

[0115] Mass spectrometry instrument: HPLC: Waters e2695 Alliance; and detector: Acquity-QDa.

[0116] The compounds described herein were purified using the preparative HPLC procedure described below.

[0117] Preparative HPLC: WATERS 2555 quaternary gradient module (maximum total flow rate: 300 mL / min, maximum pressure: 3000 psi) or Shimadzu LC-8A (maximum total flow rate: 150 mL, maximum pressure: 30 MPa), column: phenyl, 10 µm, flow rate: 75 mL / min

[0118] Mobile phase:

[0119] The purity of the compounds described herein was analyzed using one of the RP-HPLC methods described below.

[0120] HPLC Method A

[0121] Column: Xbridge peptide BEH C18 (4.6 mm x 250 mm, 3.5 μm)

[0122] Elution buffer: Mobile phase A: Buffer: Acetonitrile = 900:100 Mobile phase B: Buffer: Acetonitrile = 300:700 Buffer solution: aqueous solution of potassium dihydrogen phosphate, adjusted to pH 3.0 ± 0.1 with orthophosphate. Flow rate: 0.8 mL / min Detection: UV detection at 210 nm Column temperature: 65°C Sample tray temperature: 5°C Running time: 90 minutes

[0123] HPLC Method B

[0124] Column: YMC Pack Pro C18 (4.6 mm x 250 mm, 3.0μ)

[0125] Elution buffer: Mobile phase A: Buffer: Acetonitrile = 900:100 Mobile phase B: Buffer: Acetonitrile = 300:700 Buffer solution: aqueous solution of potassium dihydrogen phosphate, adjusted to pH 3.0 ± 0.1 with orthophosphate. Flow rate: 1.0 mL / min Detection: UV detection at 210 nm Column temperature: 50°C Sample tray temperature: 5°C Running time: 38 minutes

[0126] HPLC method C

[0127] Column: X-Select CSH C18, 130 A°, 2.5 µm, (4.6 X 150) mm

[0128] Elution buffer: Mobile phase A: Buffer: Acetonitrile = 900:100 Mobile phase B: Buffer: Acetonitrile = 300:700 Buffer solution: potassium dihydrogen phosphate aqueous solution; trimethylamine added; pH adjusted to 2.5 ± 0.1 with orthophosphate. Flow rate: 0.5 mL / min Detection: UV detection at 214 nm Column temperature: 60°C Sample tray temperature: 5°C Running time: 90 minutes

[0129] HPLC method D

[0130] Column: X-Select CSH C18, 130 A°, 2.5 µm, (4.6 X 150) mm

[0131] Elution buffer: Mobile phase A: Buffer: Acetonitrile = 900:100 Mobile phase B: Buffer: Acetonitrile = 300:700 Buffer solution: potassium dihydrogen phosphate aqueous solution; trimethylamine added; pH adjusted to 2.5 ± 0.1 with orthophosphate. Flow rate: 0.8 mL / min Detection: UV detection at 210 nm Column temperature: 60°C Sample tray temperature: 5°C Running time: 33 minutes

[0132] Preparation method

[0133] Example A: Part A-II Uncle Preparation of butyl ester

[0134] Part A-II Uncle Butyl acetate

[0135] Partial A-II was prepared using solid-phase synthesis. UncleButyl ester. In the presence of DIPEA, 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid is linked to 2-chlorotriphenylmethyl chloro resin to produce 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The Fmoc protecting group is removed by selective deblocking with piperidine, followed by coupling with Fmoc-Aib-OH in THF using DIPC and HOBt to produce 2-[2-[2-[(2-Fmoc-amino-2-methyl-propionyl)amino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The Fmoc group is removed by selective deblocking with piperidine, and the free amino group is coupled with Fmoc-Glu-OtBu using HOBt and DIPC to produce 2-[2-[2-[[2-[[(4)] S )-4-Fmoc-amino-5- Uncle Butoxy-5-oxo-pentanoyl]amino]-2-methyl-propionyl]amino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The Fmoc group of the resulting compound was selectively deblocked using piperidine, and then the free amino group was reacted with octadecanoic acid mono-resin. Uncle Butyl ester coupling yields 2-[2-[2-[[2-[[(4)] S )-5- Uncle Butoxy-4-[(18- Uncle Butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]-2-methyl-propionyl]-amino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The intermediate was then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[[2-[[(4) S )-5- Uncle Butoxy-4-[(18- Uncle [Butoxy-18-oxo-octadecanoyl]amino]-5-oxo-pentanoyl]amino]-2-methyl-propionyl]amino]ethoxy]ethoxy]acetic acid (partial A-di) Uncle Butyl ester). LCMS = m / z: 786.39 (M+H) + ).

[0136] Preparation of part of A-OSu

[0137] Part A-OSu

[0138] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting A-di Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound A-OSu.

[0139] Example B: Part B-II Uncle Preparation of butyl ester

[0140] Partial β-di-tert-butyl ester

[0141] Part B-II was prepared using a method similar to that given in Example A. Uncle Butyl ester, wherein 20-( Uncle (Butoxy)-20-oxoeicosanoic acid replacing octadecanoic acid monobutylene oxide Uncle Butyl ester, yielding 2-[2-[2-[[2-[[(4) S )-5- Uncle Butoxy-4-[(20- Uncle Butoxy-20-oxo-eicosanoyl)amino]-5-oxo-pentanoyl]amino]-2-methyl-propionyl]amino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. This intermediate is then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[[2-[[(4)] S )-5- Uncle Butoxy-4-[(20- Uncle [Butoxy-20-oxo-eicosanoyl]amino]-5-oxo-pentanoyl]amino]-2-methyl-propionyl]amino]ethoxy]ethoxy]acetic acid (partially β-di) Uncle Butyl ester). LCMS = m / z: 814.10 (M+H) + ).

[0142] Preparation of some B-OSu

[0143] Partial B-OSu

[0144] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting partial B-di Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound, part B-OSu.

[0145] Example C: Part C-II Uncle Preparation of butyl ester

[0146] Partial C-di-tert-butyl ester

[0147] Partial C-II was prepared using solid-phase synthesis. UncleButyl ester. In the presence of DIPEA, 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid is linked to a 2-chlorotriphenylmethyl chlororesin to produce 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The Fmoc protecting group is removed by selectively deblocking the amino group using piperidine, and then... right Nitrophenyl chloroformate activates the free amino group in THF and DIPEA, followed by reaction with Fmoc-aminobutyramine hydrochloride in THF:DMAc and DIPEA to produce 2-[2-[2-(4-Fmoc-aminobutyrcarbamoylamino)ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The Fmoc group is removed by selective deblocking with piperidine, and then the free amino group is coupled to Fmoc-Glu-OtBu using HOBt and DIPC to produce 2-[2-[2-[4-[[(4-aminobutyrcarbamoylamino)ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. S )-4-Fmoc-amino-5- Uncle Butoxy-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The resulting 2-[2-[2-[4-[[(4)] resin was selectively prepared using piperidine. S )-4-Fmoc-amino-5- Uncle Butoxy-5-oxo-pentanoyl]amino]-butylcarbamoylamino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl- resin deblocking, then with octadecanoic acid mono- Uncle Butyl ester coupling yields intermediate 2-[2-[2-[4-[[(4)] S )-5- Uncle Butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The intermediate was then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[4-[[(4 S )-5- Uncle Butoxy-4-[(18- Uncle [Butoxy-18-oxo-octadecanoyl]amino]-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid (partial C-di) Uncle Butyl ester). LCMS = m / z: 814.56 (M+H) + ).

[0148] Preparation of some C-OSu

[0149] Some C-OSu

[0150] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting C-di... Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound moiety C-OSu.

[0151] Example D: Part D-II Uncle Preparation of butyl ester

[0152] Part D2 Uncle Butyl acetate

[0153] Part B-II was prepared using a method similar to that given in Example C. Uncle Butyl ester, wherein 20-( Uncle (Butoxy)-20-oxoeicosanoic acid replacing octadecanoic acid monobutylene oxide Uncle Butyl ester, yielding intermediate 2-[2-[2-[4-[[5- Uncle Butoxy-4-[(20- Uncle [Butoxy-20-oxo-eicosanoyl]amino]-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The intermediate was then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[4-[[5- Uncle Butoxy-4-[(20- Uncle [Butoxy-20-oxo-eicosanoyl]amino]-5-oxo-pentanoyl]amino]butylcarbamoylamino]ethoxy]ethoxy]acetic acid (partially D-di) Uncle Butyl ester). LCMS = m / z: 843.14 (M+H) + ).

[0154] Preparation of part of D-OSu

[0155] Some D-OSu

[0156] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting partial D-di... Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound, part D-OSu.

[0157] Example E: Preparation of a portion of E-OSu

[0158] Part E-OSu

[0159] L-glutamic acid α- Uncle Butyl ester (H-Glu-OtBu) reacts with palmitic acid in the presence of IBCF and NMM to produce CH3-(CH2). 14 -C(O)-Glu-OtBu, then reacted with HOSu in the presence of IBCF and NMM to produce CH3-(CH2). 14 -C(O)-Glu(OSu)-O t Bu, then deprotected with trifluoroacetic acid to produce partial E-OSu.

[0160] Example F: Part F-II Uncle Preparation of butyl ester

[0161] Partial F-2 Uncle Butyl acetate

[0162] Partial F-II was prepared using solid-phase synthesis. Uncle Butyl ester. In the presence of DIPEA, 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid is linked to a 2-chlorotriphenylmethyl chloro resin to produce 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-chlorotriphenylmethyl resin. The Fmoc protecting group is removed by selectively deblocking the amino group with piperidine, followed by coupling with 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid in THF using DIPC and HOBt to produce {(Fmoc-aminoethoxy)-ethoxy}-acetyl-{(-aminoethoxy)-ethoxy}-acetic acid-2-chlorotriphenylmethyl resin. By selectively deblocking the Fmoc group with piperidine and coupling the free amino group with Fmoc-Glu-OtBu using HOBt and DIPC, Fmoc-Glu({(amino-ethoxy)-ethoxy}-acetyl-{(-amino-ethoxy)-ethoxy}-acetic acid-2-chlorotriphenylmethyl-resin)-OtBu is generated. The Fmoc group of the resulting compound is then selectively deblocked with piperidine, and the free amino group is then coupled with octadecanoic acid monoester. Uncle Butyl ester coupling yields 2-[2-[2-[[2-[2-[2-[5-] Uncle Butoxy-4-[(18- Uncle Butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The intermediate was then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[[2-[2-[2-[5- Uncle Butoxy-4-[(18- Uncle [Butoxy-18-oxo-octadecanoyl]amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid (partial F-di) Uncle Butyl ester). LCMS = m / z: 846.10 (M+H) + ).

[0163] Preparation of part of F-OSu

[0164] Some F-OSu

[0165] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting partial F-di... Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound, part F-OSu.

[0166] Example G: Preparation Part G

[0167] Part G-2 Uncle Butyl acetate

[0168] Partial G-II was prepared using a method similar to that given in Example F. Uncle Butyl ester, wherein 20-( Uncle (Butoxy)-20-oxoeicosanoic acid replacing octadecanoic acid monobutylene oxide Uncle Butyl ester was used to give the intermediate 2-[2-[2-[[2-[2-[2-[[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-eicosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The intermediate was then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[[2-[2-[2-[[5-tert-butoxy-4-[(20-tert-butoxy-20-oxo-eicosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetyl]amino]ethoxy]ethoxy]acetic acid (partially G-di- Uncle Butyl ester). LCMS = m / z: 874.15 (M+H) + ).

[0169] Preparation of part of G-OSu

[0170] Some G-OSu

[0171] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting G-di... Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound, G-OSu.

[0172] Example H: Preparation of partial H-di Uncle Butyl acetate

[0173] Partial H-2 Uncle Butyl acetate

[0174] Partial H-dihydrochloride was prepared using solid-phase synthesis with 2-chlorotriphenylmethyl chloride resin. Uncle Butyl ester. In the presence of DIPEA, 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid is linked to a 2-chlorotriphenylmethyl chloro resin to produce 2-[2-(2-Fmoc-aminoethoxy)ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The Fmoc protecting group is removed by selectively deblocking the amino group with piperidine, followed by coupling with Fmoc-Glu-OtBu using HOBt and DIPC to produce 2-[2-[2-[[(4S)-5- Uncle Butoxy-4-(9 H [fluorene-9-ylmethoxycarbonylamino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid-2-chlorotriphenylmethyl-resin. The Fmoc group of the resulting compound was selectively deblocked using piperidine, and then the free amino group was reacted with octadecanoic acid mono- Uncle Butyl ester coupling yields 2-[2-[2-[[(4S)-5- Uncle Butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid 2-chlorotriphenylmethyl-resin. The intermediate was then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[[(4S)-5-tert-butoxy-4-[(18-tert-butoxy-18-oxo-octadecanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid (partially H-diethyl) Uncle Butyl ester). LCMS = m / z: 700.94 (M+H) + ).

[0175] Preparation of part of H-OSu

[0176] Partial H-OSu

[0177] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting H-di... Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound, part H-OSu.

[0178] Example I: Preparation Part I

[0179] Part I-II Uncle Butyl acetate

[0180] Part I-II was prepared using a method similar to that given in Example H. Uncle Butyl ester, wherein 20-( Uncle (Butoxy)-20-oxoeicosanoic acid replacing octadecanoic acid monobutylene oxide Uncle Butyl ester, yielding intermediate 2-[2-[2-[[(4S)-5-tert-butoxy-4-[(20- Uncle Butoxy-20-oxo-eicosanoyl)amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid 2-chlorotriphenylmethyl-resin. The intermediate was then cleaved from the 2-chlorotriphenylmethyl-resin using trifluoroethanol:DCM (1:1) to give 2-[2-[2-[[(4S)-5-tert-butoxy-4-[(20- Uncle [Butoxy-20-oxo-eicosanoyl]amino]-5-oxo-pentanoyl]amino]ethoxy]ethoxy]acetic acid (partial I-di) Uncle Butyl ester). LCMS = m / z: 728.99 (M+H) + ).

[0181] Preparation of part of I-OSu

[0182] Part I-OSu

[0183] Then, in the presence of dicyclohexylcarbodiimide (DCC), the resulting partial I-di Uncle Butyl ester reacts with HOSu to produce a succinimide-protected intermediate, which is then deprotected with trifluoroacetic acid to produce the title compound, part I-OSu.

[0184] Example 12: Synthesis of Compound 12

[0185] The parent peptide was synthesized via a solid-phase method. The starting resin used for synthesis was Fmoc-Rink amide resin. Piperidine was used to selectively deblock the Fmoc-protected amino groups of the Rink amide resin, followed by coupling Fmoc-Ser(tBu)-OH with the Rink amide resin. This was achieved by using diisopropylcarbodiimide... N1,3-hydroxybenzotriazole (DIPC-HOBt) was used as a coupling agent to generate Fmoc-Ser(tBu)-Rink amide resin, completing the first cycle. Acetic anhydride and diisopropylethylamine were used to terminate / cap uncoupled amino groups at each amino acid coupling site. Piperidine was used to selectively de-block the amino groups of the Fmoc-Ser(tBu)-Rink amide resin. Then, HOBt and DIPC were used to couple Fmoc-Pro-OH to generate Fmoc-Pro-Ser(tBu)-Rink amide resin, completing the second cycle.

[0186] For the remaining 36 amino acid residues, repeat the above three steps: end capping, selectively decapping the Fmoc protection of the amino acid linked to the resin, and sequentially coupling the next amino acid residue to the Fmoc-protected amino group. The side chains of the Fmoc-protected amino acids are orthogonally protected (e.g., the hydroxyl groups of serine, tyrosine, or threonine are protected with...). Uncle Butyl (-tBu) group protection, lysine amino group protected by... Uncle The butoxycarbonyl (-Boc) and (4,4-dimethyl-2,6-dioxocyclohexyl-1-ylidene)-3-methylbutyl (IVDde) groups are protected, and the carboxylic acid group of aspartic acid or glutamic acid is protected with (- t The amide group of glutamine is protected by a triphenylmethyl (-Trt) group. Perform the above three steps, namely selective end-capping, de-blocking, and then coupling with the next Fmoc-protected amino acid, to obtain Fmoc-Aib-Gln(Trt)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-(αMethyl-Leu)-Leu-Asp(OtBu)-Lys(Boc)-Lys(IVDde)-Ala-Gln(Trt)-Aib-Ala-Phe-Ile-Glu(OtBu)-Tyr(tBu)-Leu-Leu-Glu(OtBu)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-

[0187] The resin was deblocked using piperidine, followed by the use of diisopropylcarbodiimide. N -Hydroxybenzotriazole (DIPC-HOBt) is used as a coupling agent to couple Boc-Tyr(tBu)-OH, producing Boc-Tyr(tBu)-Aib-Gln(Trt)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-(αMethyl-Leu)-Leu-Asp(OtBu)-Lys(Boc)-Lys(IVDde)-Ala-Gln(Trt)-Aib-Ala-Phe-Ile-Glu(OtBu)-Tyr(tBu)-Leu-Leu-Glu(OtBu)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-resin. The IVDde group of the peptide resin was deprotected using hydrazine hydrate, followed by the use of diisopropylcarbodiimide. N -Hydroxybenzotriazole (DIPC-HOBt) was used as a coupling agent for partial A-di Uncle The coupling of butyl ester, in the presence of the coupling agent, produces compound 12 on the resin. The resin is then cleaved and deprotected using trifluoroacetic acid with ethane-1,2-dithiol and triisopropylsilane, followed by preparative HPLC purification to obtain purified compound 12.

[0188] Mass spectrometry (LCMS): m / z = 1197.92 (MH4 4+); calculated mass = 4787.64; HPLC purity (method C): 97.4%.

[0189] Example 13: Synthesis of Compound 13

[0190] Compound 13 was prepared by a solid-state method following a similar approach to that given in Example 12, wherein after deprotection of IVDde, it is partially β-dioxanone. Uncle Coupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0191] Mass spectrometry (LCMS): m / z = 1204.89 (MH4 4+); calculated mass = 4815.52; HPLC purity (method C): 97.2%.

[0192] Example 14: Synthesis of Compound 14

[0193] Compound 14 was prepared by a solid-phase method following a similar procedure as given in Example 12, wherein (i) Fmoc-ortholeucine-OH was used instead of Fmoc-αMe-leucine-OH at position 13, and (ii) after IVDde deprotection, it was partially β-di Uncle Coupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0194] Mass spectrometry (LCMS): m / z = 1201.39 (MH4 4+); calculated mass: 4801.53; HPLC purity (method C): 98.09%.

[0195] Example 15: Synthesis of Compound 15

[0196] Compound 15 was prepared by a solid-state method following a similar approach to that given in Example 14, wherein after deprotection of IVDde, it is partially A-di Uncle Coupling of butyl ester, rather than partial β-diethyl ester. Uncle Butyl ester coupling.

[0197] Mass spectrometry (LCMS): m / z = 1194.39 (MH4 4+); calculated mass: 4773.53; HPLC purity (method C): 96.4%.

[0198] Example 16: Synthesis of Compound 16

[0199] Compound 16 was prepared by a solid-phase method following a similar method to that given in Example 12, wherein Fmoc-valine-OH was used instead of Fmoc-αMe-leucine-OH at position 13.

[0200] Mass spectrometry (LCMS): m / z = 1191.32 (MH4 4+) and calculated mass = 4761.25.

[0201] Example 17: Synthesis of Compound 17

[0202] Compound 17 was prepared by a solid-state method following a similar approach to that given in Example 16, wherein the IVDde deprotection resulted in a partial β-dioxanone. UncleCoupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0203] Example 18: Synthesis of Compound 18

[0204] Compound 18 was prepared by a solid-state method following a similar approach to that given in Example 16, wherein after deprotection of IVDde, it is partially G-di Uncle Coupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0205] Example 19: Synthesis of Compound 19

[0206] Compound 19 was prepared by a solid-state method following a similar approach to that given in Example 12, wherein after deprotection of IVDde, it is partially C-di Uncle Coupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0207] Mass (LCMS) m / z: 1205.64 (MH4 4+) and calculated mass: 4818.53.

[0208] Example 20: Synthesis of Compound 20

[0209] Compound 20 was prepared by a solid-state method following a similar approach to that given in Example 12, wherein the deprotected IVDde yielded a partially D-dioxanone compound. Uncle Coupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0210] Mass (LCMS) m / z: 1212.64 (MH4 4+) and calculated mass: 4846.53.

[0211] Example 21: Synthesis of Compound 21

[0212] The parent peptide was synthesized via a solid-phase method. The starting resin used for synthesis was Fmoc-Rink amide resin. Piperidine was used to selectively deblock the Fmoc-protected amino groups of the Rink amide resin, followed by coupling Fmoc-Ser(tBu)-OH with the Rink amide resin. This was achieved by using diisopropylcarbodiimide... N1,3-hydroxybenzotriazole (DIPC-HOBt) was used as a coupling agent to generate Fmoc-Ser(tBu)-Rink amide resin, completing the first cycle. Acetic anhydride and diisopropylethylamine were used to terminate / cap the uncoupled amino group at each amino acid coupling point. Piperidine was used to selectively de-block the amino group of the Fmoc-Ser(tBu)-Rink amide resin. Then, HOBt and DIPC were used to couple with Fmoc-Pro-OH to generate Fmoc-Pro-Ser(tBu)-Rink amide resin, completing the second cycle. Acetic anhydride and diisopropylethylamine were used to terminate the uncoupled amino group after each amino acid coupling.

[0213] For the remaining 36 amino acid residues, repeat the above three steps: end capping, selectively decapping the Fmoc protection of the amino acid linked to the resin, and sequentially coupling the next amino acid residue to the Fmoc-protected amino group. The side chains of the Fmoc-protected amino acids are orthogonally protected (e.g., the hydroxyl groups of serine, tyrosine, or threonine are protected with...). Uncle Butyl (-tBu) group protection, lysine amino group protected by... Uncle The butoxycarbonyl (-Boc) and (4,4-dimethyl-2,6-dioxocyclohexyl-1-ylidene)-3-methylbutyl (IVDde) groups are protected, and the carboxylic acid group of aspartic acid or glutamic acid is protected with (- t The amide group of glutamine is protected with a triphenylmethyl (-Trt) group. The above three steps are performed: selective end-capping, de-blocking, and then coupling with the next Fmoc-protected amino acid to obtain Fmoc-Aib-Gln(Trt)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-N-valine-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-resin.

[0214] The resin was deblocked using piperidine, followed by the use of diisopropylcarbodiimide. N -Hydroxybenzotriazole (DIPC-HOBt) is used as a coupling agent for the coupling of Boc-Tyr(tBu)-OH to produce Boc-Tyr(tBu)-Aib-Gln(Trt)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-N-valine-Leu-Asp(OtBu)-Lys(Boc)-Ile-Ala-Gln(Trt)-Lys(IVDde)-Ala-Phe-Val-Gln(Trt)-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-resin. The IVDde group of the peptide resin was deprotected using hydrazine hydrate, followed by partial A-dihydroxybenzotriazole (DIPC-HOBt) coupling with diisopropylcarbodiimide. Uncle The coupling of butyl ester yields compound 21 on the resin. Compound 21 is then purified by cleavage and deprotection of the resin using trifluoroacetic acid with ethane-1,2-dithiol and triisopropylsilane, followed by preparative HPLC purification.

[0215] Mass (LCMS) m / z: 1185.39 (MH4 4+); calculated mass: 4737.53; HPLC purity (method C): 98.5%.

[0216] Example 22: Synthesis of Compound 22

[0217] Compound 22 was prepared by a solid-state method following a similar approach to that given in Example 21, wherein after deprotection of IVDde, it is partially β-dioxanone. Uncle Coupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0218] Mass spectrometry (LCMS): m / z = 1192.41 (MH4 4+); calculated mass: 4765.60; HPLC purity (method C): 97.6%.

[0219] Example 23: Synthesis of Compound 23

[0220] Compound 23 was prepared by a solid-phase method following a similar method to that given in Example 22, wherein Fmoc-Asn(Trt)-OH was used instead of Fmoc-Gln(Trt)-OH at position 3.

[0221] Mass spectrometry (LCMS): m / z = 1189.20 (MH4 4+) and calculated mass: 4752.77.

[0222] Example 24: Synthesis of Compound 24

[0223] Compound 24 was prepared by a solid-phase method following a similar method to that given in Example 21, wherein Fmoc-Asn(Trt)-OH was used instead of Fmoc-Gln(Trt)-OH at position 3.

[0224] Mass spectrometry (LCMS): m / z = 1182.22 (MH4 4+) and calculated mass: 4724.85.

[0225] Example 26: Synthesis of Compound 26

[0226] Compound 26 was prepared by a solid-state method following a similar approach to that given in Example 12, wherein after deprotection of IVDde, it is partially G-di Uncle Coupling of butyl ester, rather than partial A-di Uncle Butyl ester coupling.

[0227] Mass spectrometry (LCMS): m / z = 1219.95 (MH4 4+) and calculated mass: 4875.76; HPLC purity (method C): 96.3%.

[0228] Biological research

[0229] Example 1: Efficacy study in db / db mice at a dose of 10 nM / kg

[0230] The effects of the compounds described herein on blood glucose, food intake, and body weight were investigated in mice. This study was conducted in a type 2 diabetic mouse (db / db) model. Animals were randomly assigned to nine treatment groups (n=6): a diabetic control group, compound 12 (10 nM / kg), compound 13 (10 nM / kg), compound 26 (10 nM / kg), compound 14 (10 nM / kg), compound 15 (10 nM / kg), compound 21 (10 nM / kg), compound 22 (10 nM / kg), and tirzepatide (10 nM / kg). Baseline blood glucose was measured in all animals. All animals were administered the test compounds subcutaneously. Blood glucose was measured at 4, 8, 12, 24, 48, 72, and 96 hours post-treatment. Delta blood glucose (mM) was calculated. Body weight change and cumulative food intake were measured at 48 and 96 hours post-treatment.

[0231] Table 3. Effects on blood glucose

[0232] Compared to the diabetic control group p<0.05, p<0.01, p<0.001, and a two-way ANOVA was subsequently performed using the Bonferroni post-test.

[0233] Table 4. Effects on body weight

[0234] Compared to the diabetic control group p<0.05, p<0.01, p < 0.001; one-way ANOVA was followed by Dunnett post-test.

[0235] Table 5. Effects on food intake

[0236] Compared to the diabetic control group p<0.05, p<0.01, p < 0.001; one-way ANOVA was followed by Dunnett post-test.

[0237] The above results indicate that the compound can effectively treat type 2 diabetes, diabetes with obesity, obesity, and hyperlipidemia.

[0238] Example 2: Oral glucose tolerance test (OGTT) in rats; single injection; 30 nM / kg dose

[0239] Animals were randomly assigned to six groups (n=4 / group): placebo control, compound 17 (30 nM / kg), compound 18 (30 nM / kg), compound 19 (30 nM / kg), compound 20 (30 nM / kg), and retaglutide (30 nM / kg). Animals were fasted for 12 hours prior to the start of the OGTT. Blood glucose was measured using a glucometer 24 hours after subcutaneous injection of the test substance or retaglutide (measurement at time 0). All animals were orally administered 2 g / kg of glucose solution. Blood glucose was measured at 10, 20, 40, 60, 90, and 120 minutes after the glucose challenge. Body weight and food intake were recorded at 12, 48, and 72 hours.

[0240] Table 6. Blood glucose AUC 24 hours after subcutaneous injection (0-120分钟) Changes

[0241] Compared to placebo control p<0.05, p<0.01, p < 0.001; one-way ANOVA was followed by Bonferroni post-test.

[0242] Table 7. Weight loss (%)

[0243] Compared to placebo control p<0.05, p<0.01, p < 0.001, and relative to retaliglutide #p < 0.05, ##p < 0.01 and ###p < 0.001; One-way ANOVA was followed by Bonferroni post-test.

[0244] Table 8. Food intake (g)

[0245] Compared to placebo control p<0.05, p<0.01, p < 0.001, and relative to retaliglutide #p < 0.05, ##p < 0.01 and ###p < 0.001; One-way ANOVA was followed by Bonferroni post-test.

[0246] Surprisingly, the inventors found that the compound exhibited weight loss comparable to that of retaglutide, but without a significant reduction in food intake. This is likely because the compound has lower potency for GCGR (relative to GLP-1R) compared to retaglutide.

[0247] Variations in individual receptor agonism in GLP-1 / GCG dual receptor agonists and GLP-1 / GIP / GCG triple receptor agonists may lead to different outcomes in terms of therapeutic efficacy and adverse effects.

[0248] To achieve optimal efficacy and minimize side effects, a delicate balance must be struck between GLP-1 / GIP / GCG receptor agonism.

[0249] Designing molecules that demonstrate enhanced therapeutic efficacy and lower side effects compared to existing therapies is a challenge. The weight loss induced by compounds 17, 18, 19, and 20 without a sharp decrease in food intake may be related to the improved efficacy of these compounds on GLP-1R relative to GCGR, which can be derived from Example 7.

[0250] Example 3: in vitro Measurement

[0251] Cell lines stably expressing GLP-1R, GIPR, or GCGR were used to assay compounds as described herein. in vitro Efficacy. Signaling in GLP-1R, GIP-R, and GCG-R involves the activation of adenylate cyclase and the production of cAMP. The Hit Hunter® cAMP assay uses a technique called Enzyme Fragment Complementation (EFC) developed by DiscoverX, with β-galactosidase (β-Gal) as the functional reporter, to monitor the activation of GLP-1R, GIPR, or GCGR via Gi and Gs secondary messenger signaling. The enzyme is split into two complementary parts: EA stands for enzyme acceptor, and ED stands for enzyme donor. ED is fused to cAMP and competes with cell-produced cAMP for binding to a cAMP-specific antibody during the assay. Active β-Gal is formed by the complementation of exogenous EA with any unbound ED cAMP. The active enzyme can then convert a chemiluminescent substrate, producing an output signal detectable on a standard microplate reader.

[0252] Three different assays were performed using cells expressing any of the three receptors. The cAMP Hunter cell line was expanded from frozen stock according to standard procedures. 20 μL of cells were seeded into white-walled 384-well microplates and incubated at 37°C for an appropriate time prior to assay. The culture medium was aspirated, and the cells were treated with 15 μL of cAMP-conjugated antibody and 5 μL of the assay compound. After incubation with the appropriate compound, an assay signal was generated by incubation with 20 μL of cAMP-ED cell lysis mixture for 1 hour, followed by incubation with 20 μL of cAMP-EA reagent for 3 hours at room temperature. Free cAMP-ED available in the system is complementary to free cAMP-EA to form active β-Gal, which reacts with the substrate to generate a chemiluminescent signal. The microplate was read using a PerkinElmer Envision™ instrument after signal generation for chemiluminescent signal detection. The amount of signal was proportional to the concentration of cAMP generated as a result of the response. Different concentrations of samples (different for different compounds) were used to generate log concentration versus % effect curves. Four-parameter logistic curves were generated, and EC50 was determined. Use the appropriate assay reference for each assay (exotropic peptide-4 for GLP-1R, GIP for GIPR, glucagon for GCGR).

[0253] Cellular cAMP assays were performed for retaglutide, compounds 12, 13, 14, 17, 18, 19 and 20, and the half-maximal effective concentrations for GLP-1R-expressing cells and GIPR-expressing cells are shown in Table 9.

[0254] Table 9.

[0255] The lower affinity of the GLP-1 receptor for the GCG receptor can reduce several side effects caused by GCGR agonism.

[0256] Glucagon agonism is diabetic and leads to increased heart rate, catabolism of amino acids and proteins, and consequently, loss of lean body mass. Therefore, the compounds of the present invention provide several benefits and a reduced profile of side effects.

Claims

1. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: X1-X2-X3-G-T-F-T-S-D-X10-S-X12-X13-L-D-X16-X17-X18-X19-X20-X21-F-X23-X24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 1) wherein: X1 is Y; X2 is Aib; X3 is Q or N; X10 is Y; X12 is I; X13 is an L or D isomer of an amino acid of the formula: X16 is K; X17 is I or K; X18 is A; X19 is Q; X20 is K, Aib, or an L or D isomer of an amino acid of the formula: X21 is A; X23 is V or I; X24 is Q or E; X25 is W or Y; X27 is I or L; X28 is A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; wherein the acid group of the C-terminal amino acid is a free carboxylic acid group or is amidated to a C-terminal primary amide; and 3. The polypeptide of claim 2, wherein the linker is selected from the group consisting of aminoethoxyethoxyacetic acid, glutamic acid, diaminobutane, Aib, and any combination thereof.

4. The polypeptide of claim 3, wherein the glutamic acid is g-glutamic acid.

5. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: X1-X2-X3-G-T-F-T-S-D-X10-S-X12-X13-L-D-X16-X17-X18-X19-X20-X21-F-X23-X24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 2) wherein: X1 is Y; X2 is Aib; X3 is Q or N; X10 is Y; X12 is I; X13 is aMe-L; X16 is K; X17 is I or K; X18 is A; X19 is Q; X20 is K, Aib, or an L or D isomer of an amino acid of the formula: X21 is A; X23 is V or I; X24 is Q or E; X25 is W or Y; X27 is I or L; X28 is A or E; X29 is G; X30 is G; X31 is P; X32 is S; X33 is S; X34 is G; X35 is A; X36 is P; X37 is P; X38 is P; and X39 is S; wherein the acid group of the C-terminal amino acid is a free carboxylic acid group or is amidated to a C-terminal primary amide; and 6. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: ​ ​ ​ ​ wherein represents the point of attachment to Leu, and wherein R is selected from C1-C6alkyl, C3-C6cycloalkylmethyl, and C3-C6cycloalkyl; ​ ​ ​ ​ ​ wherein " represents the point of attachment to Leu, and wherein R is selected from C1-C6alkyl, C3-C6cycloalkylmethyl, and C3-C6cycloalkyl; ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ with the proviso that at least one of X17 and X20 is K, and at least one of said Ks is in C 16 -C 22 fatty acid conjugation.

2. The polypeptide of claim 1, wherein K is linked to C via a linker 16 -C 22 fatty acid conjugation. ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ wherein ” denotes the point of attachment to Leu, and wherein R is selected from C1-C6alkyl, C3-C6cycloalkylmethyl, and C3-C6cycloalkyl; ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ provided that at least one of X17and X20is K, and further provided that at least one of said K comprises a side chain amino group acylated with a moiety selected from the group consisting of: NH2 Aminoethoxyethoxyacetic acid-Aib-Glu-C 16 -C 22 Fatty acid chain; Aminoethoxyethoxyacetic acid-C(O)-diaminobutane-Glu-C 16 -C 22 Fatty acid chain; Glu-C 16 -C 22 fatty acid chain; Aminoethoxyethoxyacetic acid - Aminoethoxyethoxyacetic acid - Glu-C 16 -C 22 fatty acid chain; and aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 fatty acid chain; with the proviso that when X20 is Aib, the side chain amino group is not aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 fatty acid chain or aminoethoxyethoxyacetic acid-Glu-C 16 -C 22 fatty acid chain acylated. ​ X1-X2-X3-G-T-F-T-S-D-X10-S-X12-X13-L-D-X16-X17-X18-X19-X20-X21-F-X23-X24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 3) wherein: X1is Y; X2is Aib; X3is Q or N; X10is Y; X12is I; X13is Aib, Ser(OMe), nor-V, nor-L, or aMe-L; X16is K; X17is I or K; X18is A; X19is Q; X20is K, Aib, Ser(OMe), nor-V, or nor-L; X21is A; X23is V or I; X24is Q or E; X25is W or Y; X27is E, I, or L; X28is A or E; X29is G; X30is G; X31is P; X32is S; X33is S; X34is G; X35is A; X36is P; X37is P; X38is P; and X39is S; wherein the acid group of the C-terminal amino acid is a free carboxylic acid group or is amidated to a C-terminal primary amide; and provided that at least one of X17and X20is K, and further provided that at least one of said K comprises a side chain amino group acylated with a moiety selected from the group consisting of: NH2 wherein the polypeptide is not SEQ ID NO: 7, SEQ ID NO: 23, or SEQ ID NO:

30.

7. The polypeptide of claim 6, wherein: X1is Y; X2is Aib; X3is Q or N; X10is Y; X12is I; X13is nor-V, nor-L, or aMe-L; X16is K; X17is I or K; X18is A; X19is Q; X20is K, Aib, nor-V, or nor-L; X21is A; X23is V or I; X24is Q or E; X25is W or Y; X27is I or L; X28is A or E; X29is G; X30is G; X31is P; X32is S; X33is S; X34is G; X35is A; X36, X37, and X38are each independently P; and X39is S; where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: wherein the polypeptide is not SEQ ID NO:

7.

8. The polypeptide of claim 6, wherein: X1is Y; X2is Aib; X3is Q or N; X10is Y; X12is I; X13is nor-V, nor-L, or aMe-L; X16is K; X17is K; X18is A; X19is Q; X20is Aib, nor-V, or nor-L; X21is A; X23is I; X24is E; X25is Y; X27is L; X28is E; X29is G; X30is G; X31is P; X32is S; X33is S; X34is G; X35is A; X36, X37, and X38are each independently P; and X39is S; where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: wherein the polypeptide is not SEQ ID NO:

7.

9. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: X1-X2-X3-G-T-F-T-S-D-X10-S-X12-X13-L-D-X16-X17-X18-X19-X20-X21-F-X23-X24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 3) wherein: X1is Y; X2is Aib; X3is Q or N; X10is Y; X12is I; X13is Aib, Ser(OMe), nor-V, nor-L, or aMe-L; X16is K; X17is I or K; X18is A; X19is Q; X20is K, Aib, Ser(OMe), nor-V, or nor-L; X21is A; X23is V or I; X24is Q or E; X25is W or Y; X27is E, I, or L; X28is A or E; X29is G; X30is G; X31is P; X32is S; X33is S; X34is G; X35is A; X36is P; X37is P; X38is P; and X39is S; wherein the acid group of the C-terminal amino acid is a free carboxylic acid group or is amidated to a C-terminal primary amide; and wherein the polypeptide is not SEQ ID NO: 7, SEQ ID NO: 23, or SEQ ID NO:

30. X1-X2-X3-G-T-F-T-S-D-X10-S-X12-X13-L-D-X16-X17-X18-X19-X20-X21-F-X23-X24-X25-L-X27-X28-X29-X30-X31-X32-X33-X34-X35-X36-X37-X38-X39 (SEQ ID NO: 4) wherein: X1is Y; X2is Aib; X3is Q or N; X10is Y; X12is I; X13is nor-V, nor-L, or aMe-L; X16is K; X17is I or K; X18is A; X19is Q; X20is K or Aib; X21is A; X23is V or I; X24is Q or E; X25is W or Y; X27is I or L; X28is A or E; X29is G; X30is G; X31is P; X32is S; X33is S; X34is G; X35is A; X36, X37, and X38are each independently P; and X39is S; with the proviso that at least one of X17and X20is K, and further with the proviso that the K comprises a side chain amino group acylated with a moiety selected from the group consisting of: NH2 。 10. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: Y-Aib-X3-G-T-F-T-S-D-Y-S-I-X13-L-D-K-X17-A-Q-X20-A-F-X23-X24-X25-L-X27-X28-G-G-P-S-S-G-A-P-P-P-S (SEQ ID NO: 5) wherein: X3is Q or N; X13is nor-V, nor-L, or aMe-L; X17is I or K; X20is K or Aib; X23is V or I; X24is Q or E; X25is W or Y; X27is I or L; and X28is A or E; wherein the acid group of the C-terminal amino acid is a free carboxylic acid group or is amidated to a C-terminal primary amide; and wherein the polypeptide is not SEQ ID NO:

7.

11. The polypeptide of claim 10, wherein: X3is Q; X13is aMe-L; X17is K; X20is Aib; X23is I; X24is E; X25is Y; X27is L; and X28is E.

12. The polypeptide of claim 10, wherein: X3is Q; X13is nor-V; X17is K; X20is Aib; X23is I; X24is E; X25is Y; X27is L; and X28is E.

13. The polypeptide of claim 10, wherein: X3is Q; X13is nor-L; X17is K; X20is Aib; X23is I; X24is E; X25is Y; X27is L; and X28is E.

14. The polypeptide of claim 10, wherein: X3is Q; X13is nor-V; X17is I; X20is K; X23is V; X24is Q; X25is W; X27is I; and X28is A.

15. The polypeptide of claim 10, wherein: X3is N; X13is nor-V; ​ ​ ​ ​ ​ ​ ​ with the proviso that at least one of X17and X20is K, and further with the proviso that the K comprises a side chain amino group acylated with a moiety selected from the group consisting of: NH2 ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: 。 ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: 。 ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: 。 ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ wherein the side chain amino group of K at position X20 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: 。 ​ ​ ​ X17 is I; X20 is K; X23 is V; X24 is Q; X25 is W; X27 is I; and X28 is A; wherein the side chain amino group of K at position X20 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: 。 16. A polypeptide or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: Y-Aib-X3-G-T-F-T-S-D-Y-S-I-X13-L-D-K-X17-A-Q-X20-A-F-I-E-Y-L-L-E-G-G-P-S-S-G-A-P-P-P-S (SEQ ID NO: 6) wherein: X3 is Q or N; X13 is nor-V, nor-L, or aMe-L; X17 is K; and X20 is Aib, nor-L, or nor-V; wherein the acid group of the C-terminal amino acid is a free carboxylic acid group or is amidated to a C-terminal primary amide; and where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: wherein the polypeptide is not SEQ ID NO:

7.

17. The polypeptide of claim 16, wherein: X13 is nor-V or nor-L; and X20 is Aib; where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: 。 18. The polypeptide of claim 16, wherein: X13 is aMe-L; and X20 is nor-L or nor-V; where the side chain amino group of K at position X17 is acylated with a moiety selected from the group consisting of: an amino) group is acylated with a moiety selected from the group consisting of: 。 19. An incretin analog comprising: a lysine residue comprising a fatty acid elongation group attached to the lysine epsilon-nitrogen; a peptide residue comprising the sequence Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) indirectly attached via its carboxy terminus to the lysine residue; a peptide residue having the sequence Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) indirectly attached via the amino terminus of the Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue to the carboxy group of the lysine; and a norvaline amino acid residue indirectly attached to and between the Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) residue and the lysine residue.

20. An incretin analog comprising a lysine residue comprising a group of formula (I) attached to the lysine epsilon-nitrogen, wherein formula (I) is (I) wherein U is absent or represents -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-}, wherein} is the point of attachment to W; W represents: -C(O)-CH2-O-(CH2)2-O-(CH2)2-NH-], -C(O)-NH-(CH2) 3-4 -NH-], -C(O)-C(CH3)2-NH-] or , wherein ] is the point of attachment to Y; Y is absent or represents -C(O)-(CH2)2-CH(CO2H)NH-- or -C(O)CH((CH2) x CO2H)NH-- wherein x is 1, 2 or 3 and - is the point of attachment to Z; and Z represents -C(O)-(CH2) n -COOH or -C(O)-(CH2) n -CH3, wherein n is an integer from 14 to 20; a peptide residue comprising the sequence Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) indirectly attached via its carboxy terminus to the lysine residue; a peptide residue having the sequence Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2 (SEQ ID NO: 32) indirectly linked to the carboxyl of said lysine via the amino terminus of a Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue; and a norvaline amino acid residue indirectly linked to and between the Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) residue and said lysine residue.

21. A polypeptide or an incretin analog selected from the group consisting of: 。 22. The incretin analog of claim 19 or claim 20, wherein said lysine is linked to said Gly-Thr-Phe-Thr-Ser-Asp (SEQ ID NO: 31) residue by a peptide residue comprising 10 amino acids.

23. The incretin analog of claim 19 or claim 20, wherein said lysine is linked to said Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-CONH2 (SEQ ID NO: 32) residue by a peptide residue comprising 11 amino acids.

24. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide or an incretin analog of any one of claims 1 to 22.

25. A method of treating obesity comprising administering to a patient in need of such treatment a polypeptide or an incretin analog of any one of claims 1 to 22.

26. A method of treating type 2 diabetes (T2DM) comprising administering to a patient in need of such treatment a polypeptide or an incretin analog of any one of claims 1 to 22.

27. A method of treating metabolic syndrome comprising administering to a patient in need of such treatment a polypeptide or an incretin analog of any one of claims 1 to 22.

28. A method of treating metabolic dysfunction associated steatohepatitis (MASLD) comprising administering to a patient in need of such treatment a polypeptide or an incretin analog of any one of claims 1 to 22.

29. A method of treating metabolic dysfunction associated steatohepatitis (MASH) comprising administering to a patient in need of such treatment a polypeptide or an incretin analog of any one of claims 1 to 22.

30. A method of treating a neurodegenerative disorder comprising administering to a patient in need of such treatment a polypeptide or an incretin analog of any one of claims 1 to 22.

31. A method of treating fibrosis comprising administering to a patient in need of such treatment a polypeptide or an incretin analog of any one of claims 1 to 22.

32. A method of reducing cardiovascular risk comprising administering to a patient in need of such treatment a polypeptide or incretin mimetic according to any one of claims 1 to 22.

33. A method of treating hyperlipidemia / dyslipidemia comprising administering to a patient in need of such treatment a polypeptide or incretin mimetic according to any one of claims 1 to 22.

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