GIP, GLP-1, and GLP-2 Peptide Triple Agonist

Abstract

The present invention relates to peptide triple agonists of GIPR (glucose-dependent insulinotropic polypeptide receptor), GLP-1R (glucagon-like peptide-1 receptor) and GLP-2R (glucagon-like peptide-2 receptor) and their use in medicine.

Description

[Technical field]

[0001] The present invention relates to peptide triple agonists of GIPR (glucose-dependent insulinotropic polypeptide receptor), GLP-1R (glucagon-like peptide-1 receptor) and GLP-2R (glucagon-like peptide-2 receptor) and their use in medicine. [Background technology]

[0002] Gut peptides and adipokines are essential signaling molecules involved in the control of whole-body energy homeostasis. These circulating hormones regulate various biological responses, such as hunger, satiety, and glucose uptake. In vivo experiments have established that these hormones also regulate bone metabolism, while human clinical trials have observed an association between these hormones and bone mass.

[0003] Incretins are gut hormones that help regulate carbohydrate metabolism in response to food intake. The two major incretins are glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1), both of which are secreted by intestinal epithelial cells. Intestinal glucagon-like peptide-2 (GLP-2) is co-secreted with GLP-1 upon nutrient ingestion.

[0004] Gut hormones released after meal intake, such as GIP and GLP-2, have been shown to regulate bone turnover, with GIP having a positive effect on bone and GLP-2 regulating bone homeostasis and contributing positively to bone mass.

[0005] GLP-1R may also contribute to bone metabolism.

[0006] Osteoporosis can be defined as a combination of reduced bone mass and altered bone quality, resulting in reduced bone strength with increased risk of fracture.Gastrointestinal hormones, including GIP and GLP-2, have each been implicated in bone metabolism and as potential therapeutics for treating osteoporosis.

[0007] WO2018 / 069442 and WO2020 / 169792 both disclose dual peptide agonists of GIPR and GLP-2R.

[0008] It is difficult to target specific receptors of the same receptor family, such as family B GPCRs, while not targeting others. Potent peptides that are simultaneously agonists of GIPR, GLP-1R, and GLP-2R have not been previously disclosed. Summary of the Invention

[0009] Provided herein are triple agonists that combine the properties of GIP, GLP-1, and GLP-2 receptor agonists. The inventors have designed peptide triple agonists that target GIPR, GLP-1R, and GLP-2R with unexpectedly high potency.

[0010] The triple GIPR, GLP-1R and GLP-2R agonist of the present invention induces lower internalization of GIPR compared to human GIP. This can lead to long-term activation of GIPR and therefore more sustained effect. Moreover, some of the triple GIPR, GLP-1R and GLP-2R agonists of the present invention also induce lower internalization of GLP-1R and / or GLP-2R compared to human GLP-1 and / or GLP-2. Lower internalization of any one of GIPR, GLP-1R and / or GLP-2R can lead to long-term activation and therefore more sustained and higher effect.

[0011] One aspect of the present disclosure is a method for the preparation of a compound of the sequence X 1 X 2 X 3GTFX 7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO: 42)-J In a further aspect, the present invention relates to a peptide triple agonist comprising or consisting of the following sequence: X 1 is H or Y, X 2 is A or Aib, X 3 is D or E, X 7 is I, S, or T, X 11 is S or A, X 12 is T or I, X 13 is I, Aib, or E; X 15 is D or E, X 16 is N, L, A, K, or E; X 17 is L, K, or I; X 20 is R or K, X 21 is D, E, or K; X 24 is N or E, X 28 is Q, A, or K, X 29 is T, Q, or K; X 30 is R or K, X 31 is I or G, X 32 is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29), or is omitted, However, X 7 If I, then X 2 is Aib and X 2 If A, then X 7 is S or T, the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of SEQ ID NO: 42; The peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0012] Another aspect of the disclosure relates to a peptide triple agonist according to any of the preceding claims for use as a medicament, such as for use in a method of inhibiting bone resorption and / or stimulating bone formation, and / or for use in a method of treating a bone disorder, and / or for use in a method of treating idiopathic osteoporosis, and / or for use in a method of treating osteoporosis in an individual suffering from Duchenne muscular dystrophy (DMD) or cerebral palsy. [Brief description of the drawings]

[0013] [Figure 1]The overall potency of triple peptide agonists against human GIPR, GLP-1R, and GLP-2R is shown in FIG. 1 as a radar plot. Here, compounds 20, 21, 22, 28, 30, 31, 32, 33, 34, 35 are shown in terms of their potency in Gs activation measured as cAMP accumulation, as illustrated in the radar plot. Numbers refer to -pLog EC50 values. Corresponding values ​​against the human glucagon receptor (GCGR) are also illustrated, but all have potencies below 100 nM (-pLog EC50 values ​​< 7). The outer ring corresponds to higher potency and the inner ring corresponds to lower potency. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] definition The term "affinity" refers to the strength of binding between a ligand and its receptor.

[0015] The term "agonist" in the present context refers to a peptide, as defined herein, that is capable of binding to and activating a receptor.

[0016] The term "triple agonist" refers to a peptide as defined herein that can bind and activate at least three receptors, where the at least three receptors are different receptors.In this context, a triple agonist is an agonist of GIPR, an agonist of GLP-2R, and an agonist of GLP-1R.A triple agonist as defined herein may also have agonist activity on additional receptors, whereby the triple agonist is an agonist of at least GIPR, GLP-2R, and GLP-1R.

[0017] An "amino acid residue" may be a natural or unnatural amino acid residue linked by a peptide bond or a bond other than a peptide bond. The amino acid residue may be in the D or L configuration. An amino acid residue is a residue consisting of amino terminal portions (NH2 ) and a carboxy terminal portion (COOH), at least one of which contains at least one side chain or functional group. NH 2 refers to the amino group present at the amino terminus of an amino acid or peptide, and COOH refers to the carboxy group present at the carboxy terminus of an amino acid or peptide. The generic term amino acid includes both natural and unnatural amino acids. Natural amino acids according to standard nomenclature as listed in J. Biol. Chem., 243:3552-59 (1969) and adopted in 37 C.F.R., section 1.822(b)(2) are assigned to the following groups of amino acids listed herein: Y, G, F, M, A, S, I, L, T, V, P, K, H, Q, E, W, R, D, N, and C. Unnatural amino acids are those not listed immediately above. Unnatural amino acid residues also include, but are not limited to, modified amino acid residues, L-amino acid residues, and stereoisomers of D-amino acid residues.

[0018] "Equivalent amino acid residue" refers to an amino acid residue that can replace another amino acid residue in a polypeptide without substantially changing the structure and / or functionality of the polypeptide. Thus, equivalent amino acids have similar properties such as side chain bulkiness, side chain polarity (polar or non-polar), hydrophobicity (hydrophobic or hydrophilic), pH (acidic, neutral, or basic), and side chain configuration of carbon molecules (aromatic / aliphatic). Thus, "equivalent amino acid residues" can be considered as "conservative amino acid substitutions."

[0019] Within the meaning of the term "equivalent amino acid substitution" as applied herein, in one embodiment one amino acid may be substituted for another within the group of amino acids set out herein below.

[0020] Amino acids with polar side chains (Asp, Glu, Lys, Arg, His, Asn, Gln, Ser, Thr, Pro, and Cys); amino acids with nonpolar side chains (Gly, Ala, Val, Leu, Ile, Phe, Trp, Tyr, and Met); amino acids with aliphatic side chains (Gly, Ala, Val, Leu, Ile); amino acids with cyclic side chains (Trp, His, Pro); amino acids with aromatic side chains (Phe, Tyr, Trp); acidic, e.g., negatively charged, amino acids with side chains (Asp, Glu); amino acids with basic, e.g. positively charged, side chains (Lys, Arg, His); amino acids with amide side chains (Asn, Gln); amino acids with hydroxy side chains (Ser, Thr); amino acids with sulfur-containing side chains (Cys, Met); neutral weakly hydrophobic amino acids (Pro, Ala, Gly, Ser, Thr); hydrophilic acidic amino acids (Gln, Asn, Glu, Asp); and hydrophobic amino acids (Leu, Ile, Val).

[0021] Where the L or D form (optical isomer) is not specified, it is understood that the amino acid in question may have either the naturally occurring L form (see Pure & Appl. Chem. Vol. (56(5) pp 595-624 (1984)) or the D form, such that the peptide formed may be composed of L, D, or a series of mixed L and D amino acids.

[0022] A "functional variant" of a peptide is a peptide that can perform essentially the same function as the peptide of which it is a functional variant. In particular, a functional variant can bind to the same molecule, preferably with the same affinity, as the peptide of which it is a functional variant.

[0023] A "bioactive agent" (i.e., biologically active substance / drug) is any agent, drug, compound, composition, or mixture that provides some pharmacological (often beneficial) effect that can be demonstrated in vivo or in vitro. It refers to peptide sequences as defined herein, compounds or compositions containing them, and nucleic acid constructs encoding the peptides. As used herein, the term further includes any physiologically or pharmacologically active substance that produces a local or systemic effect in an individual. As used herein, "bioactive agent" collectively refers to peptides, nucleic acid constructs encoding the peptides, and compositions containing the peptides.

[0024] As used herein, the terms "drug" and "medicine" include biologically, physiologically, or pharmacologically active substances that act locally or systemically in the human or animal body.

[0025] The terms "treatment" and "treating" as used herein refer to the management and care of a patient for the purpose of combating a condition, disease, or disorder. This term is intended to include the full range of treatments for a given condition suffered by a patient, and refers equally to curative, prophylactic or preventative therapy, and ameliorative or palliative therapy, such as administering a peptide or composition for the purpose of preventing or reducing the risk of contracting a condition, disease, or disorder, such as alleviating or relieving symptoms or complications; slowing the progression of a condition, partially halting clinical signs, disease, or disorder; curing or eliminating a condition, disease, or disorder; improving or palliating a condition or symptoms, whether detectable or undetectable, and remission (whether partial or complete); and / or preventing or reducing the risk of contracting a condition, disease, or disorder, where "preventing" or "prevention" is understood to refer to the management and care of a patient for the purpose of preventing the onset of a condition, disease, or disorder, and includes administering an active compound to prevent or reduce the risk of developing symptoms or complications. As used herein, the term "palliative" and variations thereof means that the severity and / or undesirable symptoms of a physiological condition or symptom are reduced and / or the time course of progression is slowed or prolonged compared to when a composition of the invention is not administered.

[0026] The term "individual" refers to a vertebrate, a particular member of a mammalian species, preferably a primate, including a human. As used herein, "subject" and "individual" may be used interchangeably. However, treatment of animals such as mice, rats, dogs, cats, cows, horses, sheep, and pigs is also within the scope of the invention.

[0027] "Individual in need thereof" refers to an individual who may benefit from treatment. In one embodiment, the individual in need thereof is an individual suffering from a disease, which may be a bone disorder.

[0028] A "treatment effect" or "therapeutic effect" is manifested when there is a change in the condition being treated as measured by the criteria constituting the definition of the terms "treat" and "treatment". There is a "change" in the condition being treated when there is at least a 5% improvement, preferably a 10% improvement, more preferably at least a 25%, even more preferably at least a 50%, such as at least a 75%, and most preferably at least a 100% improvement. The change can be based on an improvement in the severity of the condition being treated in an individual, or on the difference in the frequency of the improved condition in a population of individuals who have and have not been treated with a bioactive agent or a bioactive agent in combination with a pharmaceutical composition of the invention.

[0029] Treatment according to the invention may be prophylactic, ameliorative and / or curative.

[0030] A "pharmacologically effective amount," "pharmacologically effective amount," or "physiologically effective amount" of a "bioactive agent" is the amount of bioactive agent present in a pharmaceutical composition described herein required to provide a desired level of active agent in the bloodstream or at the site of action (e.g., lungs, gastric system, colorectal system, prostate, etc.) in an individual receiving treatment so as to result in the expected physiological response when such composition is administered.

[0031] As used herein, "co-administer" or "co-administration" refers to the administration of one or more agonists and a state-of-the-art pharmaceutical composition. These at least two components may be administered separately, sequentially, or simultaneously.

[0032] As used herein in connection with the peptide triple agonists of the present disclosure, "biased" refers to the peptide triple agonist being unbalanced, for example in terms of potency, activation, efficacy, and / or internalization, relative to the cognate endogenous peptide hormone for one (or more) of the three receptors GIPR, GLP-1R, and GLP-2R. The biased peptide triple agonists of the present disclosure may be beneficial for certain applications.

[0033] GIP refers to glucose-dependent insulinotropic polypeptide, also known as gastric inhibitory peptide (or polypeptide). As used herein, the abbreviation hGIP is human GIP (Uniprot Accession No. P09681). GIP is derived from a 153 amino acid proprotein and circulates as a 42 amino acid biologically active peptide (positions 52-93). It is synthesized by K cells in the duodenal and jejunal mucosa of the gastrointestinal tract.

[0034] Under physiological conditions, GIP, a 42 amino acid hormone, is degraded by the enzyme dipeptidyl peptidase 4 (DPP-4), which cleaves the GIP molecule at the third position to generate GIP3-42. GIP1-30 is generated as a result of post-translational processing. When GIP1-30 is secreted into the circulation in humans, the cleavage catalyzed by DPP-4 would result in GIP3-30.

[0035] The sequence of hGIP is YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKHNITQ (SEQ ID NO:37).

[0036] GIPR (or GIP receptor) refers to glucose-dependent insulinotropic polypeptide receptor(s). These seven-transmembrane proteins are found at least on the beta cells of the pancreas. As used herein, the abbreviation hGIPR is human GIPR (Uniprot accession number P48546).

[0037] Several physiological effects of GIP have been identified. GIP is secreted from enteroendocrine K cells following nutrient ingestion and induces insulin secretion. The amount of insulin secreted is greater when glucose is administered orally than intravenously. GIP also appears to have a profound effect on fatty acid metabolism via stimulation of lipoprotein lipase activity in adipocytes. GIP has recently been identified as a major factor in bone remodeling, and deficiency of the GIP receptor has been associated with a dramatic decrease in bone quality and subsequent increased risk of fracture.

[0038] Glucagon-like peptide-2 (GLP-2) is a 33-amino acid peptide generated in humans by specific post-translational proteolytic cleavage of proglucagon in a process that also liberates the related glucagon-like peptide-1 (GLP-1) and glucagon itself. GLP-2 is produced by enteroendocrine L-cells and various neurons of the central nervous system. Intestinal GLP-2 is co-secreted with GLP-1 upon nutrient ingestion. When administered exogenously, GLP-2 produces several effects in humans and rodents, including intestinal growth, enhanced intestinal function, reduced bone destruction, and neuroprotection. GLP-2 and related analogs have potential as therapeutic agents for short bowel syndrome, Crohn's disease, osteoporosis, and as adjunctive therapy during cancer chemotherapy.

[0039] The sequence of hGLP-2 is HADGSFSDEMNTILDNLAARDFINWLIQTKITD (sequence number 41).

[0040] GLP-2 receptor (GLP-2R) is a member of the G protein-coupled receptor superfamily. GLP-2R is expressed in the intestine and is closely related to glucagon receptor (GCGR) and GLP1 receptor (GLP-1R). As used herein, the abbreviation hGLP-2R is human GLP-2R (e.g., Uniprot accession number O95838). As used herein, the abbreviation hGLP-2 is human GLP-2 (Uniprot accession number N / A). As used herein, the abbreviation hGLP-1R is human GLP-1R (GLP-1 receptor) (e.g., Uniprot accession number P43220).

[0041] Glucagon-like peptide-1 (GLP-1) results from tissue-specific post-translational processing of proglucagon. It is produced and secreted by enteroendocrine L-cells in the intestine and by certain neurons in the nucleus of the solitary tract in the brainstem upon food consumption. In intestinal L-cells, proglucagon is processed to C-terminally amidated GLP-1 (7-36) and a small amount of C-terminally glycine-extended GLP-1 (7-37), which are released in response to a meal / glucose intake. Active GLP-1 consists of two α-helices from amino acids 13-20 and 24-35 separated by a linker region.

[0042] GLP-1 reduces blood glucose levels in a glucose-dependent manner by enhancing insulin secretion. Besides its insulin secretion-stimulating effect, GLP-1 has been associated with numerous regulatory and protective effects. Because the action of GLP-1 is preserved in patients with type 2 diabetes, considerable pharmaceutical research has been directed towards the development of GLP-1-based therapeutics. Endogenous GLP-1 is rapidly degraded, primarily by dipeptidyl peptidase-4 (DPP-4), but also by neutral endopeptidase 24.11 (NEP 24.11) and renal clearance, resulting in a half-life of approximately 2 minutes. As a result, only 10-15% of GLP-1 reaches the blood circulation intact, resulting in fasting plasma levels of only 0-15 pmol / L. To overcome this, combinations of DPP-4 inhibitors and degradation-resistant variants of GLP-1 have been developed.

[0043] The sequence of native hGLP-1 is: hGLP-1 (1 to 37): HDEFERHAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG (SEQ ID NO: 38) hGLP-1(7-36): HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR (SEQ ID NO: 39) hGLP-1(7-37): HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG (sequence number 40).

[0044] GIPR, GLP-1R, and GLP-2R peptide triple agonist The present inventors have designed novel peptides that are triple agonists of GIPR, GLP-1R, and GLP-2R, with unexpected and unprecedented high potency. These peptides are based on the sequences of hGIP and hGLP-2. Triple agonists of GIPR, GLP-1R, and GLP-2R means that the peptides bind to and / or activate at least GIPR, GLP-1R, and GLP-2R, making them potentially useful for various therapeutic applications.

[0045] The peptide is designed by combining an amino acid / strand of amino acids from a GIP peptide (SEQ ID NO:37) and an amino acid / strand of amino acids from a GLP-2 peptide (SEQ ID NO:41). Amino acid residues / strands from a GLP-1 peptide (SEQ ID NO:38) may also be present in the triple agonist of the present disclosure. These amino acids are preferably involved in one or more of the peptide's receptor binding, receptor affinity, receptor activity, and / or its agonist activity otherwise related. In some embodiments, the peptide is designed to include amino acid residues that are not derived from any of the GIP peptide (SEQ ID NO:37), GLP-1 peptide (SEQ ID NO:38), and GLP-2 peptide (SEQ ID NO:41), but may make the peptide of the present disclosure more stable and / or more potent and / or more selective compared to a peptide dual agonist that includes residues derived only from the GIP peptide (SEQ ID NO:37) and the GLP-2 peptide (SEQ ID NO:41), or only from the GIP peptide (SEQ ID NO:37), the GLP-1 peptide (SEQ ID NO:38), and the GLP-2 peptide (SEQ ID NO:41).

[0046] In one embodiment, the sequence X 1 X 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO: 42)-J and (c) providing a peptide triple agonist comprising or consisting of the sequence: X 1 is H or Y, X 2 is A or Aib, X 3 is D or E, X 7 is I, S, or T, X 11 is S or A, X 12 is T or I, X 13 is I, Aib, or E; X 15 is D or E, X 16 is N, L, A, K, or E; X 17 is L, K, or I; X 20 is R or K, X 21 is D, E, or K; X 24 is N or E, X 28 is Q, A, or K, X 29 is T, Q, or K; X 30 is R or K, X 31 is I or G, X 32 is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29), or is omitted, However, X 7 If I, then X 2 is Aib and X 2 If A, then X 7 is S or T, the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of SEQ ID NO: 42; The peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0047] In one embodiment, the sequence X 1 X 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO: 43)-J and (c) providing a peptide triple agonist comprising or consisting of the sequence: X 1 is H or Y, X 2 But, Aib, X 3 is D or E, X 7 is I, S, or T, X 11 is S or A, X 12 is T or I, X 13is I, Aib, or E; X 15 is D or E, X 16 is N, L, A, K, or E; X 17 is L, K, or I; X 20 is R or K, X 21 is D, E, or K; X 24 is N or E, X 28 is Q, A, or K, X 29 is T, Q, or K; X 30 is R or K, X 31 is I or G, X 32 is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29) or is omitted, the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of SEQ ID NO: 43; The peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0048] In one embodiment, the sequence X 1 X 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO: 44)-J and (c) providing a peptide triple agonist comprising or consisting of the sequence: X 1 is H or Y, X 2 is A or Aib, X 3 is D or E, X 7 is S or T, X 11 is S or A, X 12 is T or I, X 13 is I, Aib, or E; X 15 is D or E, X 16 is N, L, A, K, or E; X 17 is L, K, or I; X 20 is R or K, X 21 is D, E, or K; X 24 is N or E, X 28 is Q, A, or K, X 29 is T, Q, or K; X 30 is R or K, X 31 is I or G, X 32 is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29) or is omitted, the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of SEQ ID NO: 44; The peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0049] The numbering in SEQ ID NO: 42, 43, and SEQ ID NO: 44 (e.g., SEQ ID NO: 42 to 44) corresponds to the numbering of amino acid residues in the sequences, thereby X 1 is the residue at position 1 of SEQ ID NOs: 42 to 44, and X 2 is the residue at position 2, and X 3 is the residue at position 3, G is the residue at position 4, T is the residue at position 5, F is the residue at position 6, and X 7 is the residue at position 7, S is the residue at position 8, D is the residue at position 9, Y is the residue at position 10, and X 11 is the residue at position 11, and X 12 is the residue at position 12, and X 13 is the residue at position 13, L is the residue at position 14, and X 15 is the residue at position 15, and X 16 is the residue at position 16, and X 17 is the residue at position 17, A is the residue at position 18, A is the residue at position 19, and X 20 is the 20th residue, and X 21 is the residue at position 21, F is the residue at position 22, I is the residue at position 23, and X 24 is the residue at position 24, W is the residue at position 25, L is the residue at position 26, I is the residue at position 27, and X 28 is the residue at position 28, and X 29 is the residue at position 29, and X 30 is the residue at position 30, and X 31 is the residue at position 31, and X 32 is the residue at position 32 and K is the residue at position 33.

[0050] Similarly, J is a peptide numbered as a continuation of residues 1 to 33, and thus the amino acid residue or residues of J (NDWKHNITQK; SEQ ID NO:29) are numbered, for example, such that N is at position 34, D is at position 35, W is at position 36, K is at position 37, H is at position 38, N is at position 39, I is at position 40, T is at position 41, Q is at position 42, and K is at position 43.

[0051] However, in one embodiment, J may consist of only one amino acid residue, in which case, for example, the amino acid residue is at position 34, even if the amino acid residue is not N. For example, in one embodiment, J may consist of only one K, in which case, the K is at position 34.

[0052] The C-terminal region of the peptide triple agonist of the present disclosure, for example, the peptide triple agonist of SEQ ID NO: 42-44, consists of amino acid residues ranging from position 16 to the C-terminus of SEQ ID NO: 42-44. For example, the C-terminal region of the peptide triple agonist of the present disclosure, for example, the peptide triple agonist of SEQ ID NO: 42-44, consists of amino acid residues ranging from position 16 to 33 of SEQ ID NO: 42-44, for example, from position 16 to 34 of SEQ ID NO: 42-44, for example, from position 16 to 42 of SEQ ID NO: 42-44, for example, from position 16 to 43 of SEQ ID NO: 42-44.

[0053] The C-terminal region of the peptide triple agonists of the present disclosure, for example, the peptide triple agonists of SEQ ID NOs: 42-44, consists of amino acid residues 16-33 of SEQ ID NOs: 42-44 and the amino acid residue J (SEQ ID NO: 29).

[0054] Thus, in one embodiment, the peptide triple agonist of the present disclosure is modified by attaching at least one fatty acid molecule to one or more amino acid residues at any one of positions 16-33 of SEQ ID NOs:42-44 and / or at any one of positions J (SEQ ID NO:29).

[0055] Additionally, in one embodiment, the peptide triple agonist of the present disclosure is modified by attaching at least one fatty acid molecule to one or more amino acid residues at any one of positions 16-43 of SEQ ID NOs: 42-44.

[0056] In one embodiment, the peptide triple agonist of the present disclosure is modified by attaching at least one fatty acid molecule to one or more amino acid residues at any one of positions 16-33 of SEQ ID NOs:42-44.

[0057] In one embodiment, the disclosure relates to a peptide triple agonist of GIPR, GLP-1R, and GLP-2R comprising the amino acid sequence SEQ ID NOs: 42-44, or a functional variant or functional fragment thereof.

[0058] SEQ ID NOs: 42 to 44 also (H / Y)(A / Aib)(D / E)GTF(I / S / T)SDY(S / A)(I / T)(E / Aib / I)L(D / E)(E / L / K / A / N)(L / K / I)AA(R / K)(D / E / K)FI(N / E)WLI(Q / A / K)(T / Q / K)(R / K)(I / G)(T / K)KJ; or a functional variant thereof, or a functional fragment thereof.

[0059] The peptide triple agonist is an agonist of GIPR, an agonist of GLP-1R, and an agonist of GLP-2R, which means that the peptide triple agonist is an agonist of GIPR, GLP-1R, and GLP-2R. It does not exclude that the peptide triple agonist can bind to and / or activate additional receptors. However, the peptide triple agonist of the present disclosure is designed to activate GIPR, GLP-1R, and GLP-2R with high potency.

[0060] The inventors have indeed found that attachment of a fatty acid to the C-terminal region of the peptide triple agonist of the present disclosure, for example at any one of amino acid residues 16-43 of the peptides of SEQ ID NOs: 42-44, confers to the peptide the ability to bind to and / or activate GIPR, GLP-1R, and GLP-2R with high potency.

[0061] References to GIPR, GLP-2R, and GLP-1R used throughout this specification may be replaced with hGIPR, hGLP-2R, and hGLP-1R.

[0062] In one embodiment, the peptide triple agonist is a) amino acids from hGLP-2 (SEQ ID NO: 41) and / or hGIP (SEQ ID NO: 37) at the N-terminus; b. A stretch of amino acids primarily from hGIP (SEQ ID NO:37) at the central / N-terminus; c. A stretch of amino acids primarily from hGLP-2 (SEQ ID NO: 41) at the central / C-terminus, and d. optionally, amino acids primarily from hGIP (SEQ ID NO:37) at the C-terminus; Includes.

[0063] In one embodiment, amino acids 12-20 or 12-30 are primarily from hGLP-2 (SEQ ID NO:41).

[0064] In one embodiment, amino acids 5-11 or 7-11 are primarily from hGIP (SEQ ID NO:37).

[0065] In one embodiment, the amino acids at positions 7 and / or 8 are primarily from hGLP-1 (SEQ ID NO: 38). The presence of these residues may increase potency at the GLP-1R.

[0066] In one embodiment, when an amino acid is defined as "predominantly from", this means that at least 50% of the amino acids are from the peptide, such as at least 55%, such as at least 60%, for example at least 65%, such as at least 70%, for example at least 75%, such as at least 80%, for example at least 85%, such as at least 90%, for example at least 95%, such as 100% are from the peptide (i.e. from hGIP, from hGLP-1 or from hGLP-2).

[0067] In one embodiment, when amino acids are defined as "predominantly from", this means that 50-55% of the amino acids are from the peptide, such as 55-60%, for example 60-65%, such as 65-70%, for example 70-75%, such as 75-80%, for example 80-85%, such as 85-90%, for example 90-95%, such as 95-100% are from the peptide (i.e. from hGIP, from hGLP-1 or from hGLP-2).

[0068] When an amino acid is from hGIP or from hGLP-2, this means that the amino acid at a certain position of the peptide triple agonist corresponds to the amino acid at the same position of hGIP or hGLP-2. Illustratively, when the amino acid at position 1 is from GIP, this means that the amino acid is "Y". When the amino acid at position 1 is from GLP-2, this means that the amino acid is "H".

[0069] In one embodiment the peptide triple agonist comprises or consists of 20 to 43 consecutive amino acids as defined herein, such as 20 to 22 amino acids, for example 22 to 24 amino acids, such as 24 to 26 amino acids, for example 26 to 28 amino acids, such as 28 to 30 amino acids, for example 30 to 32 amino acids, such as 32 to 34 amino acids, for example 34 to 36 amino acids, for example 36 to 38 amino acids, such as 38 to 40 amino acids, for example 40 to 42, such as 38 to 43 consecutive amino acids.

[0070] In one embodiment, the peptide triple agonist comprises or consists of 20 to 43 contiguous amino acids, for example, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, or 43 contiguous amino acids.

[0071] In one embodiment, the peptide triple agonist comprises or consists of 20-33 consecutive amino acids, such as 20-21, for example 21-22, such as 22-23, for example 23-24, such as 24-25, for example 25-26, such as 26-27, for example 27-28, such as 28-29, for example 29-30, such as 30-31, for example 31-32, such as 32-33, for example 33-34, such as 34-35, for example 35-36, such as 36-37, for example 37-38, such as 38-39, for example 39-40, for example 40-41, for example 41-42, such as 42-43 consecutive amino acids.

[0072] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of 33 contiguous amino acids.

[0073] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of 34 contiguous amino acids.

[0074] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of 42 contiguous amino acids.

[0075] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of 43 contiguous amino acids.

[0076] In one embodiment, the presence of a tyrosine (Y) at position 1 results in a peptide triple agonist with increased potency for the GIPR. In one embodiment, the presence of a tyrosine (Y) at position 1 results in a peptide triple agonist with reduced potency for the GLP-1R.

[0077] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 2 is Aib, which is modified at the C-terminus of SEQ ID NOs: 42-44 by attachment of at least one fatty acid molecule.

[0078] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 2 is Aib, which is modified at the amino acid residue at position 33 of SEQ ID NOs: 42-44 by attaching at least one fatty acid molecule.

[0079] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 2 is Aib, which is modified at the amino acid residue at position 34 of SEQ ID NOs: 42-44 by attaching at least one fatty acid molecule.

[0080] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 2is Aib, which is modified at the amino acid residue at position 42 of SEQ ID NOs: 42-44 by attaching at least one fatty acid molecule.

[0081] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 2 is Aib, which is modified at the amino acid residue at position 43 of SEQ ID NOs: 42-44 by attaching at least one fatty acid molecule.

[0082] In one embodiment, the presence of a serine (S) at position 11 (in the sequence X 11 (wherein S is S) provides a peptide triple agonist with increased potency at the GLP-2R and thus a GLP-2R biased triple agonist.

[0083] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 16 is N, L, A, or E, The peptides are modified at the C-terminus of SEQ ID NOs: 42-44 by attachment of at least one fatty acid molecule.

[0084] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 16 is N, L, A, or E, The peptides are modified at the amino acid residue at position 33 of SEQ ID NOs: 42-44 by attachment of at least one fatty acid molecule.

[0085] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 16is N, L, A, or E, and the peptide is modified at the amino acid residue at position 34 of SEQ ID NOs: 42-44 by attachment of at least one fatty acid molecule.

[0086] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 16 is N, L, A, or E, and the peptide is modified at the amino acid residue at position 42 of SEQ ID NOs: 42-44 by attachment of at least one fatty acid molecule.

[0087] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 16 is N, L, A, or E, and the peptide is modified at the amino acid residue at position 43 of SEQ ID NOs: 42-44 by attachment of at least one fatty acid molecule.

[0088] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 17 is L or I.

[0089] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 20 is R.

[0090] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 7 is T. The presence of T at position 7, e.g., replacement of I at position 7 of hGIP with T, results in improved potency at GLP-1R.

[0091] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 7is S. The presence of S at position 7, e.g., replacement of I at position 7 of hGIP with S, results in improved potency against GLP-1R.

[0092] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 13 is Aib. Thus, in one embodiment, the alanine at position 13 of hGIP or the isoleucine at position 13 of hGLP-2 is substituted with 2-aminoisobutyric acid. The presence of Aib at position 13 increases potency for at least one of GIPR, GLP-1R, and GLP-2R.

[0093] In general, the presence of one or more Aib residues provides a more rigid 3D structure of the peptide triple agonist of the present disclosure, which may improve potency and selectivity for GIPR, GLP-1R, and / or GLP-2R. In one embodiment, the presence of Aib at position 2 correlates with a higher reduction in GIPR internalization, i.e., reduced GIPR internalization. In one embodiment, the presence of Aib at position 13 correlates with a higher reduction in GIPR internalization, i.e., reduced GIPR internalization. In one embodiment, the peptide triple agonist of the present disclosure comprising Aib at positions 2 and / or 13 is biased towards reduced GIPR internalization.

[0094] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 16 is N, A, or K. In one embodiment, the presence of an alanine (A) at position 16 (i.e., in the sequence, 16 A) provides peptide triple agonists with increased activity against the GIPR.

[0095] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 28 is Q.

[0096] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 29 is T.

[0097] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 30 is K.

[0098] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 2 is Aib, and X 31 is I and X 32 is T.

[0099] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 2 is Aib, and X 31 is G and X 32 is K.

[0100] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 7 is T or S, and X 31 is G and X 32 is K.

[0101] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 7 is T and X 31 is G and X 32 is K.

[0102] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NOs: 42-44, wherein the amino acid residue at position 33 is not D. Indeed, it has been found that substitution of D at position 33 of GLP-2 with K or another amino acid residue is advantageous, resulting in improved potency against GIPR and / or GLP-2R.

[0103] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NO: 42-44, wherein X 31 X 32 is selected from the group consisting of IT, GK, GT, IK, and IT. 31 X 32 is IT. In one embodiment, X 31 X 32 is the goalkeeper.

[0104] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequences SEQ ID NOs: 42-44, wherein J is omitted.

[0105] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequence SEQ ID NOs: 42-44, wherein J is ND, NDW, KDW, NDW, NDK, NDWK, NDWKH, NDWKHN, NDWKHNI, NDWKHNIT, NDWKHNITQ, and AEWKHAITQ, or a variant containing one amino acid substitution.

[0106] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequences SEQ ID NOs: 42-44, wherein J is K.

[0107] In one embodiment, the peptide triple agonist of the present disclosure comprises or consists of the amino acid sequences SEQ ID NOs: 42-44, wherein J is selected from the group consisting of NDWKHNITQ, NDWKHNITQK.

[0108] In one embodiment, the peptide triple agonist of the present disclosure comprises: Compound 1:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 1), Compound 2: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 2), Compound 3: H-Aib-DGTFISDYATILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 3), Compound 4:H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 4), Compound 5:H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-NH 2 (SEQ ID NO:5), Compound 6:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 6), Compound 7:H-Aib-DGTFISDYSTILENLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 7), Compound 8:H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKITK-NH 2 (SEQ ID NO: 8), Compound 9:H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKITK-NH 2 (SEQ ID NO: 9), Compound 10:H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 10), Compound 11:H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 11), Compound 12:H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 12), Compound 13:H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 13), Compound 14:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGKK-NH 2 (SEQ ID NO: 14), Compound 15:H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 15), Compound 16:Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 16), Compound 17:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 1), Compound 18: HADGTFTSDYSTILDNLAAKDFINWLIQTKGKK-OH (SEQ ID NO: 17), Compound 19: HADGTFTSDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 18), Compound 20: HADGTFTSDYSIILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 19), Compound 21: HADGTFTSDYSTILD-KLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 20), Compound 22: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 21), Compound 23: HADGTFTSDYSTILDKKAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 22), Compound 24: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 23), Compound 25: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 24), Compound 26: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 25), Compound 27: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 26), Compound 28: H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-OH (SEQ ID NO: 27), HADGTFISDYSTILDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 45), HADGTFISDYST-Aib-LDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 46), HADGTFISDYSTILDNKAARDFINWLIQTKGKK-OH (SEQ ID NO: 47), HADGTFISDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 48), HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 49), and HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 50), or a functional variant thereof, wherein the functional variant comprises one or two individual amino acid substitutions.

[0109] In one aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: Compound 1:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 1), Compound 2: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 2), Compound 3: H-Aib-DGTFISDYATILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 3), Compound 4:H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 4), Compound 5:H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-NH 2 (SEQ ID NO:5), Compound 6:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 6), Compound 7:H-Aib-DGTFISDYSTILENLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 7), Compound 8:H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKITK-NH 2 (SEQ ID NO: 8), Compound 9:H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKITK-NH 2 (SEQ ID NO: 9), Compound 10:H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 10), Compound 11:H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 11), Compound 12:H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 12), Compound 13:H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 13), Compound 14:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGKK-NH 2 (SEQ ID NO: 14), Compound 15:H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 15), Compound 16:Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 16), Compound 17:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 1), Compound 18: HADGTFTSDYSTILDNLAAKDFINWLIQTKGKK-OH (SEQ ID NO: 17), Compound 19: HADGTFTSDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 18), Compound 20: HADGTFTSDYSIILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 19), Compound 21: HADGTFTSDYSTILD-KLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 20), Compound 22: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 21), Compound 23: HADGTFTSDYSTILDKKAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 22), Compound 24: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 23), Compound 25: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 24), Compound 26: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 25), Compound 27: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 26), Compound 28: H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-OH (SEQ ID NO: 27), Compound 30: BB-153 HADGTFISDYSTILDKLAARDFINWLIQTKGKK-OH, Compound 31:BB-154 HADGTFISDYST-Aib-LDKLAARDFINWLIQTKGKK-OH, Compound 32: BB-155 HADGTFISDYSTILDNKAARDFINWLIQTKGKK-OH, Compound 33: BB-157 HADGTFISDYSTILDNLAARDFINWLIQTKGKK-OH, Compound 34: BB-159 HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-OH, Compound 35:BB-160 HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-OH, or a functional variant thereof, wherein the functional variant comprises one to three individual amino acid substitutions, The peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of the above sequences, The peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0110] In one aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: Compound 1: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 1), Compound 2: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 2), Compound 3: H-Aib-DGTFISDYATILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 3), Compound 4: H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 4), Compound 5: H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-NH2 (SEQ ID NO:5), Compound 6: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 6), Compound 7: H-Aib-DGTFISDYSTILENLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 7), Compound 8: H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKITK-NH2 (SEQ ID NO: 8), Compound 9: H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKITK-NH2 (SEQ ID NO: 9), Compound 10: H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 10), Compound 11: H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 11), Compound 12: H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 12), Compound 13: H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 13), Compound 14: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGKK-NH2 (SEQ ID NO: 14), Compound 15: H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKITK-NH2 (SEQ ID NO: 15), Compound 16: Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 16), Compound 17: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH2 (SEQ ID NO: 1), Compound 18: HADGTFTSDYSTILDNLAAKDFINWLIQTKGKK-OH (SEQ ID NO: 17), Compound 19: HADGTFTSDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 18), Compound 20: HADGTFTSDYSIILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 19), Compound 21: HADGTFTSDYSTILD-KLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 20), Compound 22: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 21), Compound 23: HADGTFTSDYSTILDKKAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 22), Compound 24: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 23), Compound 25: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 24), Compound 26: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 25), Compound 27: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 26), Compound 28: H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-OH (SEQ ID NO: 27), Compound 30: BB-153 HADGTFISDYSTILDKLAARDFINWLIQTKGKK-OH, Compound 31:BB-154 HADGTFISDYST-Aib-LDKLAARDFINWLIQTKGKK-OH, Compound 32: BB-155 HADGTFISDYSTILDNKAARDFINWLIQTKGKK-OH, Compound 33: BB-157 HADGTFISDYSTILDNLAARDFINWLIQTKGKK-OH, Compound 34: BB-159 HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-OH, and Compound 35:BB-160 HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-OH, or a functional variant thereof, said functional variant comprising one to three individual amino acid substitutions, with the proviso that the amino acid residue at position 2 is Aib and / or the amino acid residue at position 7 is T or S; The peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of the above sequences, The peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0111] In one embodiment, the peptide triple agonist of the present disclosure is C-terminally amidated (-NH 2 ).

[0112] In one embodiment, the C-terminus of the peptide triple agonist of the present disclosure is a carboxylic acid.

[0113] In one embodiment, a functional variant of a peptide triple agonist has at least 60% sequence identity to the peptide triple agonist, such as at least 70% sequence identity, for example at least 75% sequence identity, such as at least 80% sequence identity, for example at least 85% sequence identity, such as at least 90% sequence identity, for example at least 95% sequence identity, such as at least 97% sequence identity.

[0114] In one embodiment, a functional variant of a peptide triple agonist has 60-65% sequence identity to the peptide triple agonist, such as 65-70% sequence identity, for example 70-75% sequence identity, for example 75-80% sequence identity, for example 80-85% sequence identity, for example 85-90% sequence identity, for example 90-95% sequence identity, for example 95-99% sequence identity, for example 99-100% sequence identity. "Identity" and "sequence identity" may be used interchangeably herein.

[0115] In one embodiment, the functional variant comprises one or more amino acid substitutions, for example 1 to 8 amino acid substitutions, for example 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 7, or 7 to 8 amino acid substitutions.

[0116] In one embodiment, the functional variant comprises one amino acid substitution, two amino acid substitutions, three amino acid substitutions, four amino acid substitutions, or five amino acid substitutions.

[0117] In one embodiment, the amino acid substitutions are conservative amino acid substitutions. In one embodiment, the functional variant comprises one or more conservative amino acid substitutions.

[0118] Conservative substitutions (or synonymous substitutions) are substitutions of amino acids whose side chains have similar biochemical properties and therefore do not affect the function of the peptide.

[0119] Identity between amino acid sequences can be calculated using well-known algorithms such as BLOSUM 30, BLOSUM 40, BLOSUM 45, BLOSUM 50, BLOSUM 55, BLOSUM 60, BLOSUM 62, BLOSUM 65, BLOSUM 70, BLOSUM 75, BLOSUM 80, BLOSUM 85, or BLOSUM 90, or by simple comparison of specific amino acids present in corresponding positions in the two peptide sequences being compared. Homology can be used as a synonym for identity / sequence identity.

[0120] Conservative substitutions may be introduced at any one or more positions of a peptide according to the present disclosure so long as the variant remains functional. However, it may be desirable to introduce non-conservative substitutions (non-synonymous substitutions) at one or more positions.

[0121] Any amino acid defined herein may be in the L or D configuration. If nothing is specified, reference to the L isomer is preferably intended.

[0122] The standard and / or non-standard amino acids may be linked by peptide bonds (to form a linear peptide chain) or by non-peptide bonds (e.g., via variable side chains of the amino acids). Preferably, the amino acids of the present disclosure are linked by peptide bonds.

[0123] The terms "peptide" and "isolated peptide" may be used interchangeably herein. The terms "variant" and "functional variant" may be used interchangeably herein. The terms "fragment" and "functional fragment" may be used interchangeably herein. When reference is made herein to a "peptide", this term will include both a reference to the peptide itself, and also a reference to the peptide for the uses defined herein.

[0124] In one embodiment, the peptide is non-naturally occurring.

[0125] In one embodiment, the peptide is synthetic.

[0126] In one embodiment, the peptide is an isolated peptide.

[0127] In one embodiment, the peptide is a labeled peptide, such as a radiolabeled peptide or a fluorescently labeled peptide.

[0128] In another embodiment, variants as defined herein include sequences in which an alkyl amino acid is substituted for an alkyl amino acid, an aromatic amino acid is substituted for an aromatic amino acid, a sulfur-containing amino acid is substituted for a sulfur-containing amino acid, a hydroxy-containing amino acid is substituted for a hydroxy-containing amino acid, an acidic amino acid is substituted for an acidic amino acid, a basic amino acid is substituted for a basic amino acid, and / or a dibasic monocarboxylic amino acid is substituted for a dibasic monocarboxylic amino acid.

[0129] The term peptide also encompasses post-translational modifications introduced by chemical or enzymatic catalysis as known in the art, including acetylation, phosphorylation, methylation, glucosylation, glycation, amidation, hydroxylation, deimination, deamidation, carbamylation, and sulfation of one or more amino acid residues, as well as proteolytic modifications by known proteinases, including lysosomal cathepsins, and also calpains, secretases, and matrix metalloproteinases.

[0130] Functional equivalents of peptides may also include chemical modifications such as those made by ubiquitination, labeling (e.g., with radionuclides, various enzymes, etc.), pegylation (derivatization with polyethylene glycol), or insertion (or substitution by chemical synthesis) of amino acids such as ornithine that are not normally present in human proteins (non-proteinogenic).

[0131] Stereo-analogous compounds may be created to mimic key portions of peptide structures. This may be accomplished by modeling and chemical design techniques known to those skilled in the art. For example, the amino terminus of a di-arginine peptide backbone may be modified with esterification and other alkylation to mimic a tetrapeptide structure. It will be understood that all such sterically similar constructs fall within the scope of the present invention. Peptides with N- and C-terminal alkylation and esterification are also encompassed within the present invention.

[0132] A contiguous or consecutive peptide sequence is a linear stretch of consecutive amino acids linked by peptide bonds. Contiguous amino acid sequence and consecutive amino acid sequence are used interchangeably herein.

[0133] At least one fatty acid molecule attached The peptide triple agonists of the present disclosure are characterized in that they are modified by the attachment of at least one fatty acid molecule at an amino acid residue at the C-terminal region of the disclosed amino acid sequence. Indeed, the attachment of a fatty acid to the C-terminal region of the disclosed amino acid sequence, for example at any one of positions 16-43 of the disclosed sequence, results in a triple peptide agonist of hGIPR, hGLP-1R, and hGLP-2R that has high potency against one or more of hGIPR, hGLP-1R, and hGLP-2R.

[0134] In one embodiment, the peptide triple agonist of the present disclosure is modified by attaching at least one fatty acid molecule to a lysine (K) at any one of the positions in the C-terminal region of SEQ ID NO:42, 43, and SEQ ID NO:44 (e.g., SEQ ID NOs:42-44).

[0135] In one embodiment, the peptide triple agonist of the present disclosure is modified by attachment of at least one fatty acid molecule at any one of positions 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, or 43 of SEQ ID NOs: 42-44.

[0136] In one embodiment, the peptide triple agonist of the present disclosure is modified by attaching at least one fatty acid molecule at any one of positions 16, 17, 20, 33, 34, 37, or 43 of SEQ ID NOs: 42-44.

[0137] In one embodiment, the peptide triple agonist of the present disclosure is modified by attaching at least one fatty acid molecule to one or more amino acid residues at any one of positions 16, 17, and 20-43 of SEQ ID NOs: 42-44.

[0138] In one embodiment, the peptide triple agonists of the present disclosure are modified by attachment of at least one fatty acid molecule at one or more amino acid residues at position 16 of SEQ ID NOs: 42-44. Attachment of a fatty acid at position 16 appears to be particularly suitable for obtaining GIPR / GLP-2R / GLP-1R triple agonists with high potency against all three receptors.

[0139] In one embodiment, the peptide triple agonist of the present disclosure is modified by attachment of at least one fatty acid molecule at one or more amino acid residues at position 17 of SEQ ID NOs:42-44.

[0140] In one embodiment, the peptide triple agonist of the present disclosure is modified by attachment of at least one fatty acid molecule at one or more amino acid residues at position 20 of SEQ ID NOs:42-44.

[0141] In one embodiment, the peptide triple agonist of the present disclosure is modified by attachment of at least one fatty acid molecule at one or more amino acid residues at position 43 of SEQ ID NOs:42-44.

[0142] In one embodiment, the peptide triple agonist of the present disclosure is modified by the attachment of at least one fatty acid molecule at the C-terminus of SEQ ID NOs:42-44.

[0143] In one embodiment, attachment of one or more fatty acids at the C-terminus of a peptide triple agonist of the present disclosure at residues 33 or 34 results in a peptide triple agonist with reduced activation of the GLP-2R. In contrast, attachment of one or more fatty acids at the C-terminus of a peptide triple agonist of the present disclosure at residues 42 or 43 results in a peptide triple agonist with increased activation of the GLP-2R.

[0144] In one embodiment of the present disclosure, the fatty acid molecule is a straight chain fatty acid.

[0145] In one embodiment of the present disclosure, the fatty acid molecule is a branched fatty acid.

[0146] In one embodiment of the present disclosure, the fatty acid molecule is a monoacyl fatty acid molecule containing one fatty acid.

[0147] In one embodiment of the present disclosure, the fatty acid molecule is a diacyl fatty acid molecule.

[0148] In one embodiment of the present disclosure, the fatty acid molecule is 3 (CH 2 ) 8 CO-(Capryl, C10), CH 3 (CH 2 ) 10 CO-(Lauryl, C12), CH 3 (CH 2 ) 12 CO-(myristoyl, C14), CH 3 (CH 2 ) 14 CO-(palmitoyl, C16), CH 3 (CH 2 ) 16 CO-(Stearyl, C18) and CH 3 (CH 2 ) 18 CO-(arachidyl, C20).

[0149] In one embodiment of the present disclosure, the fatty acid molecule is 3 (CH2 ) 8 CO-(decanoyl, C10), HOOC-CH 3 (CH 2 ) 10 CO-(dodecanoyl, C12), HOOC-CH 3 (CH 2 ) 12 CO-(1-tetradecanoyl, C14), HOOC-CH 3 (CH 2 ) 14 CO-(hexadecanoyl, C16), HOOC-CH 3 (CH 2 ) 15 CO-(15-carboxy-pentadecanoyl, C17), HOOC-CH 3 (CH 2 ) 16 CO-(octadecanoyl, C18), HOOC-CH 3 (CH 2 ) 17 CO-(17-carboxy-heptadecanoyl, C19), HOOC-CH 3 (CH 2 ) 18 CO-(eicosanoyl, C20) and HOOC-CH 3 (CH 2 ) 19 It contains two acyl groups individually selected from the group consisting of CO-(19-carboxy-nonadecanoyl, C21).

[0150] In one embodiment, the fatty acid molecule is C16 (palmitoyl).

[0151] In one embodiment of the disclosure, the fatty acid molecule is attached directly to the amino acid residue, eg, without the presence of a spacer or linker.

[0152] In one embodiment of the present disclosure, the fatty acid molecule is attached to the amino acid residue via a spacer or linker.

[0153] In one embodiment, the spacer is a hydrophilic linker. In one embodiment, the spacer is a hydrophilic linker of a non-natural amino acid.

[0154] In one embodiment, the spacers are repeats of individual spacer moieties. In one embodiment, the spacers are repeats of the same spacer moiety. In one embodiment, the spacers are repeats of different spacer moieties.

[0155] In one embodiment, the fatty acid molecule is attached to the amino acid residue via a spacer in such a manner that a carboxyl group of the spacer forms an amide bond with an amino group of the fatty acid molecule.

[0156] In one embodiment, the spacer is one or more amino acids selected from the group consisting of succinic acid, Lys, Glu, Asp; 4-Abu, y-aminobutyric acid, one or more of γ-aminobutanoyl (γ-aminobutyric acid), γ-glutamyl (γ-glutamic acid), β-asparagyl, β-alanyl, and glycyl; [γ-Glutamic acid-8-amino-3,6-dioxaoctanoic acid] n (γGlu-AEEAc n ) (wherein n is an integer from 1 to 50, for example, 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, 9 to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 20, 20 to 25, 25 to 30, 30 to 35, 35 to 40, 40 to 45, or 45 to 50); The compound comprises one or more moieties individually selected from the group consisting of:

[0157] In one embodiment of the present disclosure, the spacer or linker comprises yGlu.

[0158] In one embodiment of the disclosure, the spacer comprises or consists of yGlu or yGlu-yGlu.

[0159] In some embodiments of the present disclosure, the peptide triple agonists disclosed herein are acylated, which in some embodiments renders the peptide triple agonists selective agonists of GIPR and GLP-2R rather than GLP-1R, while retaining their agonistic potency as well as increasing half-life and in vivo stability.

[0160] In one embodiment, the peptide triple agonist of the present disclosure comprises: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 1), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-OH (SEQ ID NO:2); H-Aib-DGTFISDYATILDNLAARDFINWLIQTKIT-K(C10)-OH (SEQ ID NO: 3), H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 4), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO:5), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 6), H-Aib-DGTFISDYSTILENLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 7), H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 8), H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 9), H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 10), H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 11), H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 12), H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 13), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGK-K(C10)-NH2 (SEQ ID NO: 14), H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 15), Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH2 (SEQ ID NO: 16), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(γGlu-C10)-NH2 (SEQ ID NO: 1), HADGTFTSDYSTILDNLAA-K(2×AEEA-γGlu-C18-diacid)-DFINWLIQTKGKK-OH (SEQ ID NO: 17), HADGTFTSDYSTILDNLAARDFINWLIQTKGK-K(γGlu-C18-diacid)-OH (SEQ ID NO: 18), HADGTFTSDYSIILD-K(2×AEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 19), HADGTFTSDYSTILD-K(2×AEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 20), HADGTFTSDYST-Aib-LD-K(2xAEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 21), HADGTFTSDYSTILDK-K (2xAEEA-γGlu-C18-diacid)-AARDFINWLIQTKGKKNDWKHNITQ-OH SEQ ID NO: 22), HADGTFTSDYSTILDKLAARDFINWLIQTKGK-K(2×AEEA-γGlu-C18-diacid)-NDWKHNITQ-OH (SEQ ID NO: 23), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDW-K(2×AEEA-γGlu-C18-diacid)-HNITQ-OH (SEQ ID NO: 24), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-K(2×AEEA-γGlu-C18-diacid)-OH (SEQ ID NO: 25), HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ-K (2×AEEA-γGlu-C18-diacid)-OH SEQ ID NO: 26), and H-Aib-DGTFISDYSTILDALAARDFINWLIQTKIT-K(2×AEEA-γGlu-C18-diacid)-OH (SEQ ID NO: 27), HADGTFISDYSTILDK-(2xAEEA-yGlu-C18-diacid)-LAARDFINWLIQTKGKK-OH (SEQ ID NO: 45), HADGTFISDYST-Aib-LDK-(2xAEEA-yGlu-C18-diacid)-LAARDFINWLIQTKGKK-OH (SEQ ID NO: 46), HADGTFISDYSTILDNK-(2xAEEA-yGlu-C18-diacid)-AARDFINWLIQTKGKK-OH (SEQ ID NO: 47), HADGTFISDYSTILDNLAARDFINWLIQTKGKK-(2×AEEA-yGlu-C18-diacid)-OH (SEQ ID NO: 48), HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-(2×AEEA-yGlu-C18-diacid)-OH (SEQ ID NO: 49), HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-(2×AEEA-yGlu-C18-diacid)-OH (SEQ ID NO: 50), or a functional variant thereof, wherein the functional variant comprises one or two individual amino acid substitutions.

[0161] Peptide triple agonist activity In one embodiment, a functional variant or fragment retains the same biological activity or performance as the native peptide or the peptide from which it is derived.

[0162] In one embodiment, the peptide triple agonist (including functional variants or fragments thereof) comprises: a. Binding to GIPR, GLP-1R, and GLP-2R, and / or b. activation of GIPR, GLP-1R, and GLP-2R, and / or c. Stimulation of GIPR activation, GLP-1R activation, and GLP-2R activation, such as GIPR-mediated, GLP-1R-mediated, and GLP-2R-mediated cAMP production, and / or d. inhibiting bone resorption, and / or e. stimulating bone formation; One or more of the following are possible:

[0163] In one embodiment, the peptide triple agonist (including functional variants or fragments thereof) is capable of reducing GIPR internalization compared to native hGIP.

[0164] This effect may be desirable because it may lead to sustained pharmacological effects over long periods of treatment (weeks, months, years) and may not lead to desensitization of the GIPR, which is discussed in Sameer et al. Mol Cell Biol. 2014 Oct 1;34(19):3618-29. doi:10.1128 / MCB.00256-14.

[0165] In one embodiment, the peptide triple agonist (including functional variants or fragments thereof) is capable of reducing GLP-1R internalization compared to native hGLP-1.

[0166] In one embodiment, the peptide triple agonist (including functional variants or fragments thereof) is capable of reducing GLP-2R internalization compared to native hGLP-2.

[0167] Thus, the peptide triple agonists of the present disclosure may be biased towards one receptor, e.g. - some peptide triple agonists of the present disclosure may be capable of reducing internalization of only one of GIPR, GLP-1R, and GLP-2R compared to native hGIP, hGLP-1, hGLP-2, e.g., they may be capable of reducing internalization of only GIPR, or only GLP-1R, or only GLP-2R, and thus may be biased towards GIPR or GLP-1R or GLP-2R; - Some peptide triple agonists of the present disclosure may be able to reduce internalization of two of the GIPR, GLP-1R, and GLP-2R, e.g., they may be able to reduce internalization of the GIPR and GLP-1R, but they may not be able to reduce internalization of the GLP-2R, and therefore may be biased towards the GLP-2R; Some peptide triple agonists of the present disclosure may be able to reduce internalization of all three GIPRs, GLP-1R, and GLP-2R, and thus such peptide triple agonists of the present disclosure are unbiased.

[0168] In one embodiment, the peptide triple agonists and functional variants or fragments thereof are full agonists of GIPR, GLP-1R, and GLP-2R.

[0169] In one embodiment, the peptide triple agonist can bind to and activate GIPR. In some embodiments, the GIPR is human GIPR (Uniprot accession number P48546). In one embodiment, the peptide triple agonist can bind to and activate GLP-1R. In some embodiments, the GLP-1R is human GLP-1R (Uniprot accession number P43220). In one embodiment, the peptide triple agonist can bind to and activate GLP-2R. In some embodiments, the GLP-2R is human GLP-2R (Uniprot accession number O95838).

[0170] The activity of the peptide triple agonists of the present disclosure can be assessed experimentally using a cAMP assay, such as that described in the Examples section of this specification. Using such a cAMP assay, EC 50 and E max , as well as other relevant parameters, can be determined.

[0171] In one embodiment, the peptide triple agonist provided herein comprises: a. an efficacy (E) that is at least 65% of the efficacy of native human GIP to activate human GIPR, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GIP to activate human GIPR max Activating human GIPR at b. an efficacy (E) that is at least 65% of the efficacy of native human GLP-2 to activate human GLP-2R, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GLP-2 to activate human GLP-2R max Activating the human GLP-2R at a concentration of 0.1 mM, a. an efficacy (E) that is at least 65% of the efficacy of native human GLP-1 to activate human GLP-1R, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GLP-1 to activate human GLP-1R max Activating human GLP-1R at is possible.

[0172] In one embodiment, the peptide triple agonist provided herein comprises: - activates the GIPR with the same or increased potency as native GIP activates the GIPR, - activates GLP-1R with the same or increased potency as native GLP-1 activates GLP-1R, and - activates GLP-2R with the same or increased potency as native GLP-2 activates GLP-2R; is possible.

[0173] In one embodiment, the peptide triple agonist provided herein comprises: a EC of 10 nM or less against human GIPR 50 having b. EC of 50 nM or less for human GLP-2R 50 and c. EC for human GLP-1R of 12 nM or less, e.g., 10 nM or less, e.g., 9 nM or less 50 has.

[0174] In one embodiment, the peptide triple agonist provided herein comprises: a. An EC for human GIPR of 10 nM or less, such as 9 nM or less, for example 8 nM or less, such as 7 nM or less, for example 6 nM or less, such as 5 nM or less, for example 4 nM or less, such as 3 nM or less, for example 2 nM or less, such as 1 nM or less 50 having b. EC for human GLP-2R of 50 nM or less, such as 40 nM or less, such as 30 nM or less, such as 25 nM or less, such as 20 nM or less, such as 15 nM or less, such as 10 nM or less 50 and c. an EC for human GLP-1R of 12nM or less, such as 10nM or less, for example 9nM or less, such as 8nM or less, for example 7nM or less, such as 6nM or less, for example 5nM or less, such as 4nM or less, for example 3nM or less 50 has.

[0175] Pharmaceutical / Medical Use One aspect of the present disclosure is to provide peptide triple agonists, nucleic acid constructs encoding peptide triple agonists, delivery vehicles comprising nucleic acid constructs encoding peptide triple agonists, as defined elsewhere herein, and compositions comprising peptide triple agonists, for use as pharmaceuticals.

[0176] Thus, one aspect of the present disclosure is to provide a peptide triple agonist as defined herein for use in a method of inhibiting bone resorption and / or stimulating bone formation.

[0177] It is also an aspect of the present disclosure to provide a peptide triple agonist as defined herein for use in a method of inhibiting bone resorption and / or stimulating bone formation.

[0178] There is also provided the use of a peptide triple agonist as defined herein for the manufacture of a medicament for inhibiting bone resorption and / or stimulating bone formation.

[0179] Also provided is a method of inhibiting bone resorption and / or stimulating bone formation, the method comprising administering a therapeutically effective amount of a peptide triple agonist as defined herein to an individual in need thereof.

[0180] A further aspect of the present disclosure is to provide a peptide triple agonist as defined herein for use in a method for treating bone disorders. In one embodiment, the bone disorder is associated with increased bone resorption and / or decreased bone formation. In one embodiment, the bone disorder is associated with insufficient or reduced bone density.

[0181] Also provided is a peptide triple agonist as defined herein for use in a method of treating (including treating, preventing and alleviating) a bone disorder.

[0182] Also disclosed is a peptide triple agonist as disclosed herein for use in a method of treating a bone disorder.

[0183] Bone mineral density or bone mineral density (BMD) is the amount of bone mineral in bone tissue. The concept is the amount of mineral matter per volume of bone (related to density in a physical sense), but clinically it is measured by a surrogate measure according to the optical density per square centimeter of bone surface during imaging studies. Bone mineral density measurement is used in clinical medicine as an indirect indicator of osteoporosis / osteopenia and fracture risk. It is measured by a procedure called densitometry. There is a statistical association between bone mineral density deficiency and a higher probability of fracture. Bone mineral density measurement is used to screen people for osteoporosis risk and to identify those who may benefit from measures to improve bone strength.

[0184] The T-score is a suitable measure for screening for osteoporosis. It is the bone mineral density (BMD) at that site compared to the mean value of young normal reference subjects. The World Health Organization criteria are as follows: Normal is a T score of -1.0 or greater. Osteopenia is defined as -1.0 to -2.5 Osteoporosis is defined as a bone mineral density below -2.5, meaning 2.5 standard deviations below the mean value for young normal reference subjects.

[0185] In one embodiment, bone disorder is associated with a T-score below -1.0, such as between -1.0 and -2.5, such as below -2.5.

[0186] In one embodiment there is provided the use of a peptide triple agonist as defined herein for the manufacture of a medicament for treating a bone disorder.

[0187] Also provided is a method of treating a bone disorder, the method comprising administering a therapeutically effective amount of a peptide triple agonist as defined herein to an individual in need thereof.

[0188] The individual in need as referred to herein is an individual who may benefit from the administration of triple agonist peptide. Such an individual may suffer from or be at risk of suffering from bone disorder. The individual may be any human, male or female, infant, middle-aged or elderly. The disorder that is treated or prevented in an individual may be related to the age of the individual, the general health of the individual, the medicine used to treat the individual, and whether the individual has a history of disease or disorder that may have caused or has caused bone density disorder.

[0189] In one embodiment, the bone disorder is selected from the group consisting of osteopenia, osteoporosis, severe osteoporosis, postmenopausal osteoporosis, idiopathic osteoporosis, osteomalacia, rickets, osteitis fibrosa cystic (OFC), and Paget's disease of bone.

[0190] In one embodiment, the bone disorder is osteopenia.

[0191] In one embodiment, the bone disorder is osteoporosis.

[0192] In one embodiment, the bone disorder is postmenopausal osteoporosis.

[0193] In one embodiment, the bone disorder is idiopathic osteoporosis, such as juvenile idiopathic osteoporosis.

[0194] A further aspect of the present disclosure is to provide a peptide triple agonist as defined herein for use in a method for treating osteoporosis, such as idiopathic osteoporosis in individuals with Duchenne muscular dystrophy (DMD) or cerebral palsy.

[0195] In one embodiment, the bone disorder is a secondary disorder, such as a secondary disorder in an individual with Duchenne muscular dystrophy (DMD) or cerebral palsy.

[0196] In one embodiment, the bone disorder is a secondary disorder, such as a secondary disorder in an individual receiving steroids, such as glucocorticoid treatment, and / or immobilization.

[0197] Idiopathic osteoporosis refers to the development of osteopenia and fractures with minimal or no trauma in otherwise healthy individuals who are not postmenopausal and who have no other identifiable secondary causes of osteoporosis.

[0198] Idiopathic osteoporosis refers to the development of osteopenia and fractures with minimal or no trauma in otherwise healthy individuals who are not postmenopausal and who have no other identifiable secondary causes of osteoporosis.

[0199] Also disclosed is a peptide triple agonist selected from the group consisting of SEQ ID NO:XXX, or a functional variant or functional fragment thereof, for use in a method of treating a bone disorder selected from the group consisting of osteopenia, osteoporosis, severe osteoporosis, osteomalacia, rickets, osteitis fibrosa cystic (OFC), and Paget's disease of bone.

[0200] In one embodiment, there is provided a peptide triple agonist selected from the group consisting of SEQ ID NOs: 42-44, or a functional variant or functional fragment thereof, for use in a method for treating osteopenia.

[0201] In one embodiment, there is provided a peptide triple agonist selected from the group consisting of SEQ ID NOs: 42-44, or a functional variant or functional fragment thereof, for use in a method of treating osteoporosis.

[0202] In one embodiment, there is provided a peptide triple agonist selected from the group consisting of SEQ ID NOs: 42-44, or a functional variant or functional fragment thereof, for use in a method of treating osteoporosis in an individual with Duchenne muscular dystrophy (DMD) or cerebral palsy.

[0203] Combination therapy Also, an aspect is to provide a peptide triple agonist as defined herein for use in combination with a further active pharmaceutical ingredient, which in one embodiment is useful for treating a bone disorder, such as a bone disorder associated with reduced bone density.

[0204] In one embodiment, the further active pharmaceutical ingredient is selected from the group consisting of bisphosphonates including alendronic acid (Fosamax), risedronic acid (Actonel, Atelvia, Benet), ibandronate (Boniva), zoledronic acid (Reclast, Aclasta, Zometa), etidronic acid (Didronel), pamidronic acid (Aredia / Pamimed), tiludronic acid (Skelid); estrogen replacement therapy; hormone therapy; raloxifene (Evista); calcitonin (Fortical and Miacalcin), denosumab (Prolia); teriparatide (Forteo); hormone mimetics including vitamin D (alphacalcidol or calcitriol); calcium or phosphorus supplements.

[0205] In one embodiment, the further active pharmaceutical ingredient increases the half-life of the peptide triple agonist. In one embodiment, the further active pharmaceutical ingredient is an inhibitor of dipeptidyl peptidase 4 (DPP-4 / DDP-IV inhibitor) or a gliptin. Examples include sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, dutogliptin, and omarigliptin (MK-3102).

[0206] Nucleic acid constructs encoding triple agonist peptides In one embodiment, there is provided a nucleic acid construct that encodes the triple agonist peptide defined herein.In one embodiment, said nucleic acid construct may continuously express said peptide for a long period of time, for example at least 1 month, for example at least 2 months, for example at least 3 months, for example at least 4 months, for example at least 5 months, for example at least 6 months, for example at least 7 months, for example at least 8 months, for example at least 9 months, for example at least 12 months.

[0207] Thus, one aspect is to provide a nucleic acid construct encoding a peptide triple agonist as defined herein.

[0208] In one embodiment, a nucleic acid construct is provided encoding a peptide triple agonist of SEQ ID NOs: 42-44, or a functional variant or functional fragment thereof, or any peptide triple agonist disclosed herein.

[0209] Also, an aspect is to provide a nucleic acid construct encoding a triple agonist peptide as defined herein for use in a method of treating a bone disorder.

[0210] By nucleic acid construct is understood a genetically engineered nucleic acid. The nucleic acid construct may be a non-replicating linear nucleic acid, a circular expression vector, or an autonomously replicating plasmid. The nucleic acid construct may contain several elements, such as, but not limited to, a gene or a fragment thereof, a promoter, an enhancer, a terminator, a poly-A tail, a linker, a polylinker, an operative linker, a multiple cloning site (MCS), a marker, a stop codon, an internal ribosome entry site (IRES), and a host homologous sequence for integration, or other defined elements. It is understood that the nucleic acid construct according to the present invention may contain a subset of all of the above elements or any combination thereof.

[0211] Methods for engineering nucleic acid constructs are well known in the art (see, e.g., Molecular Cloning: A Laboratory Manual, Sambrook et al., eds., Cold Spring Harbor Laboratory, 2nd Edition, Cold Spring Harbor, NY, 1989). Additionally, nucleic acid constructs according to the present invention may be synthesized without a template or obtained from various commercial suppliers (e.g., Genscript Corporation).

[0212] In one embodiment, the nucleic acid construct is a naked DNA construct comprising a sequence encoding the triple agonist peptide.

[0213] Delivery Vehicle Also, an embodiment provides a nucleic acid construct as described herein above, contained within a delivery vehicle. A delivery vehicle is an entity by which a nucleotide sequence or a polypeptide or both can be transported from at least one medium to another. A delivery vehicle is generally used for expression of a sequence encoded within the nucleic acid construct and / or for intracellular delivery of the construct or the polypeptide encoded therein.

[0214] In one embodiment, a delivery vehicle is provided comprising a nucleic acid construct as defined herein. The delivery vehicle may be selected from the group consisting of RNA-based vehicles, DNA-based vehicles / vectors, lipid-based vehicles (such as liposomes), polymer-based vehicles (such as cationic polymer DNA carriers), colloidal gold particles (coatings), and virus-derived DNA or RNA vehicles or vectors.

[0215] Non-viral delivery methods include physical approaches (carrier-free delivery) and chemical approaches (synthetic vector-based delivery).

[0216] Physical approaches, including needle injection, gene gun, jet injection, electroporation, ultrasound, and hydrodynamic delivery, use physical forces, which may be electrical or mechanical, to penetrate cell membranes and facilitate intracellular gene transfer.

[0217] Examples of chemical delivery vehicles include, but are not limited to, biodegradable polymer microspheres, lipid-based formulations such as liposomal carriers, liposomes, calcium salts, or cationically charged molecules such as dendrimers, lipopolysaccharides, polypeptides, and polysaccharides.

[0218] Another embodiment comprises a vector, herein referred to as a viral vector (i.e., non-viral), as a delivery vehicle.The viral vector according to the present invention is made of modified viral genome, i.e., the actual DNA or RNA that forms the viral genome, and is introduced in naked form.Therefore, any coat structure that surrounds the viral genome, made from viral or non-viral proteins, is not part of the viral vector.

[0219] The virus from which the viral vector is derived may be selected from the non-exhaustive group of adenovirus, retrovirus, lentivirus, adeno-associated virus, herpes virus, vaccinia virus, foamy virus, cytomegalovirus, Semliki Forest virus, pox virus, RNA viral vectors and DNA viral vectors. Such viral vectors are well known in the art.

[0220] In one embodiment, the viral vector may be selected from the group consisting of adenovirus, lentivirus, adeno-associated virus (AAV), and recombinant adeno-associated virus (rAAV). In a preferred embodiment, the viral vector is a therapeutic rAAV vector, such as a therapeutic rAAV vector.

[0221] Adenovirus is a group of viruses that contain double-stranded DNA.Adenovirus can be genetically modified to make it replication-incompetent or conditionally replication-incompetent.In this form, as adenovirus construct or adenovector, they can be used as gene delivery vehicle for vaccination or gene therapy.

[0222] Recombinant cells Another aspect relates to a cell comprising a nucleic acid construct as defined herein. Such recombinant cells can be used as a tool for in vitro research, as a delivery vehicle for the nucleic acid construct, or as part of a gene therapy program. The nucleic acid construct can be introduced into the cell by techniques well known in the art, including microinjection of DNA into the nucleus of the cell, transfection, electroporation, lipofection / liposome fusion, and particle bombardment. Suitable cells include autologous and non-autologous cells, and may include xenogeneic cells.

[0223] Preparation method (peptide) The triple agonist peptides defined herein may be prepared by any method known in the art, for example by standard peptide preparation techniques including solution synthesis or Merrifield-type solid phase synthesis.

[0224] In one embodiment, the peptide according to the present invention is synthetically produced or generated.Methods for synthetic generation of peptides are well known in the art.Detailed explanations and practical advice for generating synthetic peptides can be found in Synthetic Peptides: A User's Guide (Advances in Molecular Biology), Grant GA ed., Oxford University Press, 2002, or Pharmaceutical Formulation: Development of Peptides and Proteins, Frokjaer and Hovgaard eds., Taylor and Francis, 1999.

[0225] In one embodiment, the peptide or peptide sequence of the invention is produced synthetically, in particular by the Sequence Assisted Peptide Synthesis (SAPS) method, by solution synthesis, by solid phase peptide synthesis (SPPS) such as Merrifield type solid phase synthesis, by recombinant techniques (production by a host cell comprising a first nucleic acid sequence encoding the peptide operably linked to a second nucleic acid capable of directing expression in said host cell) or by enzymatic synthesis, which are well known to those skilled in the art.

[0226] Peptides may be synthesized either batchwise on an automated peptide synthesizer using the Na-amino protecting group and 9-fluorenylmethyloxycarbonyl (Fmoc) or tert-butyloxycarbonyl (Boc) as suitable general protecting groups for the side chain functionalities.

[0227] After purification, such as by reverse-phase HPLC, the peptides may be further processed to obtain, for example, C-terminally or N-terminally modified isoforms. Methods for terminal modifications are well known in the art.

[0228] For example, the following method for conjugation of fatty acids can be used: After removal of the final N-terminal N-Fmoc, the N-terminal amino group is acylated by treatment with 3 equivalents of 1,16-hexadecanedioic acid in the presence of 3 equivalents of HATU and 6 equivalents of DIPEA. Alternatively, lysine with a Dde-protected side chain is used for solid-phase peptide synthesis at the acylation position. After the peptide chain is assembled, the N-terminal amino group is capped by 3 equivalents of Boc2O in the presence of 6 equivalents of DIPEA. The protecting group Dde is then removed by treatment with 2% hydrazine / DMF (v / v). Alternatively, 3 equivalents of palmitoyl chloride in the presence of 6 equivalents of DIPEA can be used for palmitoylation. Alternatively, 3 equivalents of 1,16-hexadecanedioic acid can be activated by HATU / DIPEA and then conjugated to the side chain of lysine. As a further alternative, Fmoc-γ-Glu(tBu)-OH or Fmoc-4-Abu-OH or Fmoc-β-Ala-OH or Fmoc-AEEAc-OH are conjugated to the side chain of Lys using standard protocols for DIC / HOBT. The N-terminal amino group of the unnatural amino acid is then palmitoylated by treatment with 3 equivalents of palmitoyl chloride in the presence of 6 equivalents of DIPEA.

[0229] Pharmaceutical Compositions and Formulations While the triple agonist peptides defined herein can be administered as raw chemicals (peptides), it may be preferable to present them in the form of a pharmaceutical formulation, which may be referred to as a pharmaceutical composition, a pharma- ceutically acceptable composition, or a pharma- ceutically safe composition.

[0230] Therefore, also provided is a pharmaceutical formulation comprising the triple agonist peptide defined herein, or a pharma- ceutically acceptable salt or ester thereof, and a pharma- ceutically acceptable carrier, excipient, and / or diluent. Pharmaceutical formulations can be prepared by conventional techniques, for example, as described in Remington: The Science and Practice of Pharmacy 2005, Lippincott, Williams & Wilkins.

[0231] Pharmaceutically acceptable salts of the peptide compounds, where they can be prepared, are also intended to be included. These salts will be acceptable for their application in pharmaceutical use. This means that the salts will retain the biological activity of the parent compound and that they will have no untoward or detrimental effects in their application and use in treating disease.

[0232] Pharmaceutically acceptable salts are prepared in standard manner: if the parent compound is a base, it is treated with an excess of an organic or inorganic acid in a suitable solvent; if the parent compound is an acid, it is treated with an inorganic or organic base in a suitable solvent.

[0233] The peptide compounds may be administered in effective amounts in the form of their alkali metal or alkaline earth metal salts, either concurrently, simultaneously or together with pharma- ceutically acceptable carriers or diluents, especially and preferably in the form of pharmaceutical compositions thereof, whether by the oral, rectal or parenteral (including subcutaneous) route.

[0234] Examples of pharma- ceutically acceptable acid addition salts include those derived from mineral acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid, and sulfuric acid, and organic acids such as tartaric acid, acetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, succinic acid, p-toluenesulfonic acid, and arylsulfonic acids.

[0235] The pharma- ceutically acceptable salts of the peptides of the invention are preferably in a solution having a physiologically acceptable pH, ie, a solution containing the peptide salt preferably has a pH that is acceptable for clinical use.

[0236] There is provided herein a composition comprising a peptide triple agonist as defined herein, or a multimeric compound comprising said peptide triple agonist, or a nucleic acid construct comprising / encoding said peptide triple agonist.

[0237] Administration and Dosage In one embodiment of the present disclosure, the peptide or the nucleic acid construct encoding the peptide, or the composition comprising the peptide defined herein, is administered to an individual in need of treatment in a pharmacologic or therapeutically effective dose. The required dose varies depending on the specific drug composition used, the route of administration, and the specific subject being treated, which depends on the severity and type of disorder, as well as the subject's weight and general condition. Those skilled in the art will also recognize that the optimal number and interval of individual dosages will be determined by the nature and extent of the pathology being treated, the form, route, and site of administration, and the specific patient being treated, and that such optimal conditions can be determined by conventional techniques. Those skilled in the art will also recognize that the optimal course of treatment, i.e., the number of daily doses of the compound given over a specified number of days, can be ascertained using conventional course-of-treatment determination tests.

[0238] In one embodiment the peptide or composition is administered at least once a day, such as once a day, such as twice a day, such as three times a day, such as four times a day, such as five times a day.

[0239] Doses may also be administered at intermittent intervals, or at intervals such that a dose is not administered daily. Rather, one or more doses may be administered every 2 days, 3 days, 4 days, 5 days, 6 days, every week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, or intervals within these ranges (such as every 2-4 weeks, or every 4-6 weeks).

[0240] In one embodiment, the peptide or composition is administered once a week. Such an administration profile can ensure constant exposure to the effects of the peptide triple agonist of the present disclosure.

[0241] Those skilled in the art will appreciate that if the number of daily administrations is increased, the dose to be administered in each administration can be decreased accordingly.Similarly, if the duration of each administration is decreased, the dosage can be increased accordingly.

[0242] Route of administration It will be appreciated that the preferred route of administration will depend on the general condition and age of the subject being treated, the nature of the condition being treated, the location within the body of the tissue being treated, and the active ingredient selected.

[0243] whole body treatment For systemic treatment, the route of administration can introduce the peptide, a nucleic acid construct encoding the peptide, or a composition containing the peptide into the bloodstream so as to ultimately target the desired site of action.

[0244] Such routes of administration may be any suitable route, such as enteral routes (including oral, rectal, nasal, pulmonary, buccal, sublingual, transdermal, intracisternal, and intraperitoneal administration), and / or parenteral routes (including subcutaneous, intramuscular, intrathecal, intracerebral, intravenous, and intradermal administration).

[0245] Parenteral Administration Parenteral administration is any administration route that is not oral / enteral, whereby the drug avoids first-pass degradation in the liver. Thus, parenteral administration includes any injection and infusion, such as intravenous, intramuscular, or subcutaneous administration, for example, bolus injection or continuous infusion. In addition, parenteral administration includes inhalation and topical administration.

[0246] Thus, the compound may be administered topically across any mucous membrane of the animal to which the biologically active substance is to be administered, such as the nasal, vaginal, ocular, oral, genital, pulmonary, gastrointestinal, or rectal mucosa, preferably the nasal or oral mucosa, and thus parenteral administration may also include buccal, sublingual, nasal, rectal, vaginal, and intraperitoneal administration, as well as pulmonary and bronchial administration by inhalation or installation. Agents may also be administered topically across the skin.

[0247] Local treatment The peptide, or a nucleic acid construct encoding said peptide, or a composition comprising a peptide as defined herein, may in one embodiment be used as a local treatment, i.e. directly introduced into the site(s) of action. Thus, it may be applied directly to the skin or mucous membrane, or it may be injected into the site of action, for example into a terminal artery that leads directly into or into a diseased tissue.

[0248] Kit-of-parts The present invention also relates to a kit-of-parts comprising one or more of the above mentioned agents (peptides, nucleic acid constructs or compositions) and at least one additional or further component.

[0249] The kit-of-parts, in one embodiment, comprises one or more of the agents defined herein for the treatment, prevention, or alleviation of bone disorders, osteoporosis, and osteopenia. The kits defined herein allow for simultaneous, sequential, or separate administration of the active agent according to the invention and / or one or more second active ingredients described elsewhere herein. EXAMPLES

[0250] Example 1. Peptide triple agonists of GIPR, GLP-2R, and GLP-1R Materials and Methods material All applied peptides were synthesized by WuXi AppTec, Shanghai, China, with a purity of at least 95% as determined by HPLC analysis, and the correct molecular weight was confirmed using mass spectrometry. Human GIPR, GLP-2R, and GLP-1R applied in the in vitro experiments can be purchased from Origene, Rockville, USA. GIPR fused with a SNAP tag at the human N-terminus was custom produced by Cisbio, Codolet, France.

[0251] To improve the potency of the triple agonists, C10, C12, or C16, or a C16-diacid acyl group was added to the 16, 17, 20, 30, 33, 37, or C-terminal amino acid residue (C-terminus) as provided in Table 1.

[0252] result: [Table 1] TIFF2024546994000003.tif239162TIFF2024546994000004.tif76162

[0253] Example 2. Efficacy method COS-7 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) 1885 supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine, 180 units / ml penicillin, and 45 μg / ml streptomycin at 37 °C for 1 h at 20 °C in 10% CO. 2 and cultured at 37 °C. Prior to the experiments, transient transfection of COS-7 cells was performed using the calcium phosphate precipitation method as previously described ( van der Velden et al., 2021 ).

[0254] The day after transfection, transiently transfected COS-7 cells expressing either human GIPR, GLP-2R, or GLP-1R were seeded in white 96-well plates at a density of 35.000 cells / well. The next day, the assay was started by washing with HEPES-buffered saline (HBS), followed by an incubation step with assay buffer (HBS supplemented with 1 mM isobutyl-1-methylxanthine (IBMX)) for 30 min at 37°C. Ligand was then added and incubated for another 30 min at 37°C. After ligand incubation, the HitHunter® cAMP assay (Eurofins DiscoverX, Fremont, 150 USA) was performed according to the manufacturer's instructions. Luminescence was measured by a PerkinElmer™ EnVision 2014 multilabel reader (PerkinElmer, Waltham, MA). Potency (EC 50 Efficacy (E) was determined by nonlinear regression using GraphPad Prism 9 (San Diego, California, USA). Sigmoidal curves were fitted logistically. max Values) were determined by normalization to the highest applied concentration of the endogenous ligand for each receptor.

[0255] result From Tables 2, 3, and 4, it can be seen that potent GIPR / GLP-2R / GLP-1R triple agonists can be obtained by attaching fatty acid molecules at positions in the C-terminal region, such as positions 16, 17, 20, 33, 37, and the most C-terminal amino acid. For comparison, see, for example, compound No. 29, where lipidation at position 11 in the N-terminal region leads to little or no GLP-1R activity.

[0256] Attachment of a fatty acid at the C-terminus of compounds with a 1-33 or 1-34 backbone leads to reduced GLP-2R activation (see, e.g., compound 19). The same loss of activation is not observed for compounds with a fatty acid attached to the C-terminus at positions 42 or 43 of compounds with longer backbones (1-43 or 1-44) (see, e.g., compounds 24-26).

[0257] Substitution of aspartic acid (D) with glutamic acid (E) at position 3 appears to reduce potency at all three receptors GIPR, GLP-2R, and GLP-1R, thus resulting in a less potent GIPR / GLP-2R / GLP-1R triple agonist (see, e.g., compound 11). Similarly, substitution of isoleucine (I) with glutamic acid (E) at position 13 appears to reduce potency at all three receptors GIPR, GLP-2R, and GLP-1R, thus resulting in a less potent GIPR / GLP-2R / GLP-1R triple agonist (see, e.g., compound 13).

[0258] Substitution of histidine (H) with tyrosine (Y) at position 1 appears to increase activation of GIPR while simultaneously decreasing GLP-1R activation (see, e.g., compound 16).

[0259] The threonine at position 7 appears to be well tolerated by all three receptors, GIPR, GLP-2R, and GLP-1R, and therefore specifically increases GLP-1R activity, resulting in a potent GIPR / GLP-2R / GLP-1R triple agonist.

[0260] A serine (S) at position 11 appears to increase potency towards the GLP-2R, thus resulting in a potent GIPR / GLP-2R / GLP-1R triple agonist with a GLP-2R bias.

[0261] The attachment of a fatty acid at position 16 appears to be particularly suitable for obtaining a GIPR / GLP-2R / GLP-1R triple agonist with high potency at all three receptors. The attachment of a fatty acid at position 17 (see, for example, compound 32) appears to cause a loss of GIP potency (>10-fold) in cAMP as well as a small loss of GLP-2R activation compared to the attachment of a fatty acid at position 16 (see, for example, compound 30), while no loss is seen for GLP-1R activation.

[0262] Alanine at position 16 appears to be particularly suitable for increasing GLP-1R activity to obtain a GIPR / GLP-2R / GLP-1R triple agonist.

[0263] A peptide containing the sequence NDWKHNITQK (sequence number Z) (the C-terminal portion (34-42) of hGIP) appears to be highly suitable for obtaining a potent GIPR / GLP-2R / GLP-1R triple agonist.

[0264] C-terminal carboxylation (-COOH) appears to increase GIPR and GLP-1R activation while decreasing GLP-2R activation (see, for example, compound 2).

[0265] [Table 2] TIFF2024546994000006.tif171162

[0266] [Table 3] TIFF2024546994000008.tif227162TIFF2024546994000009.tif125162

[0267] [Table 4] TIFF2024546994000011.tif225162TIFF2024546994000012.tif83162

[0268] Example 3: Internalization method HEK293A cells were cultured in DMEM-GlutaMAX™-I supplemented with 10% FBS, 180 units / mL penicillin, and 45 μg / mL streptomycin at 37 °C for 24 h at 37 °C, with 5% CO 2 and cultured at 37° C. For receptor internalization assays, the cells were transfected using Lipofectamine 2000 (Thermo Fischer Scientific, Massachusetts, USA) according to the manufacturer's instructions.

[0269] On the day of transfection, HEK293A cells transiently expressing human SNAP-tagged GIPR, human SNAP-tagged GLP-1R, or human SNP-tagged GLP-2R were seeded in white 384-well plates at a density of 15.000 cells / well. The next day, the medium was removed and new fresh culture medium was added. On the third day, the assay was performed. First, cells were labeled with 100 nM Tag-Lite SNAP-Lumi4-Tb (donor) in OptiMEM for 60 min at 37 °C. Then, cells were incubated in internalization buffer (1 mM CaCl 2 The cells were washed four times with HBBS supplemented with 1 mM MgCl2, 1 mM MgCl2, and 20 mM HEPES, followed by the addition of 100 μM pre-warmed fluorescein-O'-acetate (acceptor). Plates were placed in a 37°C incubator for 5 min prior to ligand stimulation to allow temperature adjustment before plate reading. Cells were then stimulated with 37°C pre-warmed ligand solution and internalization was measured every 3 min for 60 min at 37°C in a PerkinElmer™ EnVision 2014 multilabel reader (PerkinElmer, Waltham, MA). AUC values ​​of the sigmoidal curves up to a concentration of 1 μM were calculated similarly.

[0270] result In general, the data show that the triple agonists tested induced, to different degrees, less internalization of the GIPR than native GIP and less internalization of the GLP-2R than native GLP-2. Some of the triple agonists tested also induced less internalization of the GLP-1R than native GLP-1, while other triple agonists tested induced slightly more internalization than native GLP-1.

[0271] Reduced internalization positively correlates with reduced recruitment of beta-arrestin2.

[0272] Conjugation of fatty acids to the C-terminus at position 33 of the triple agonist peptides of the present disclosure correlated with low overall internalization and bias towards GIPR receptors, with very low or absent internalization of GLP-1R and GLP-2R. See, for example, Compounds Nos. 1-17, 19, 28, 33-35 (some data reported in Tables 5-7). In contrast, Compounds Nos. 26 and 27, which are C-terminally lipidated at position 43, caused a higher degree of GIPR internalization.

[0273] The data also show that lipidation at position 16 of compounds having a 1-42 or 1-43 scaffold, e.g., compounds 20, 21, 22, resulted in unbiased internalization to the GLP-1 and GLP-2 receptors (comparable to native GLP-1 and GLP-2), but biased internalization to the GIPR (reduced internalization compared to native GIP).

[0274] The data also show that lipidation at position 16 of compounds with the 1-33, 1-34 scaffolds (see, e.g., compounds 30, 31, 32) also resulted in biased internalization towards GLP-1R, but less bias towards GLP-2R. Without being bound by theory, it is believed that i) shorter peptide scaffolds create a higher degree of bias towards one of the GIPR, GLP-1R and / or GLP-2R relative to the endogenous agonist GIP than longer peptide scaffolds, and ii) C-terminal lipidation was associated with the greatest bias (in terms of loss of internalization).

[0275] The presence of Aib at position 13 confers a slightly greater impediment (greater bias) to internalization for GIPR (see, for example, compound 26 vs. compound 27 and compound 30 vs. compound 31).

[0276] The presence of Aib at position 2, e.g., compound 28, resulted in increased internalization with a relative AUC of 58% of endogenous GIP compared to 39% for compounds 35 and 34. However, they are biased due to lower internalization compared to the endogenous agonist GIP.

[0277] [Table 5]

[0278] [Table 6]

[0279] [Table 7]

[0280] Example 4: Sequence Overview K(γGlu-C16) provided herein should be read as fatty acid C16 being attached to the side chain amino group of lysine via the yGlu linker. Thus, the yGlu linker is directly attached to the amino group on the side chain of lysine, and the fatty acid is directly attached to yGlu. The same applies to other fatty acids and linkers listed in parentheses herein below.

[0281] Compound 1:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 1), Compound 2: H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 2), Compound 3: H-Aib-DGTFISDYATILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 3), Compound 4:H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 4), Compound 5:H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-NH 2 (SEQ ID NO:5), Compound 6:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 6), Compound 7:H-Aib-DGTFISDYSTILENLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 7), Compound 8:H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKITK-NH 2 (SEQ ID NO: 8), Compound 9:H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKITK-NH 2 (SEQ ID NO: 9), Compound 10:H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 10), Compound 11:H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 11), Compound 12:H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 12), Compound 13:H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 13), Compound 14:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGKK-NH 2 (SEQ ID NO: 14), Compound 15:H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 15), Compound 16:Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 16), Compound 17:H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK-NH 2 (SEQ ID NO: 1), Compound 18: HADGTFTSDYSTILDNLAAKDFINWLIQTKGKK-OH (SEQ ID NO: 17), Compound 19: HADGTFTSDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 18), Compound 20: HADGTFTSDYSIILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 19), Compound 21: HADGTFTSDYSTILD-KLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 20), Compound 22: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 21), Compound 23: HADGTFTSDYSTILDKKAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 22), Compound 24: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 23), Compound 25: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 24), Compound 26: HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 25), Compound 27: HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQK-OH (SEQ ID NO: 26), Compound 28: H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK-OH (SEQ ID NO: 27), Compound 29: H-Aib-DGTFISDY-KTILDNLAARDFINWLIQTKITK-OH (SEQ ID NO: 28), Compound 30: HADGTFISDYSTILDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 45), Compound 31: HADGTFISDYST-Aib-LDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 46), Compound 32: HADGTFISDYSTILDNKAARDFINWLIQTKGKK-OH (SEQ ID NO: 47), Compound 33: HADGTFISDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 48), Compound 34: HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 49), Compound 35: HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 50), NDWKHNITQK (SEQ ID NO:29) NDWK (SEQ ID NO:30) NDWKH (SEQ ID NO:31) NDWKHN (SEQ ID NO:32) NDWKHNI (SEQ ID NO: 33) NDWKHNIT (SEQ ID NO: 34) NDWKHNITQ (SEQ ID NO: 35) AEWKHAITQ (SEQ ID NO: 36) YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKHNITQ (SEQ ID NO: 37) HDEFERHAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG (SEQ ID NO: 38) HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR (SEQ ID NO: 39) HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG (SEQ ID NO: 40) HADGSFSDEMNTILDNLAARDFINWLIQTKITD (SEQ ID NO: 41) HX 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO:42) HX 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO:43) HX 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO:44)

[0282] References Van der Velden et al.2021.GLP-1 Val8:A Biased GLP-1R Agonist with Altered Binding Kinetics and Impaired Release of Pancreatic Hormones in Rats

[0283] Terms 1. Arrays X 1 X 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO: 42)-J A peptide triple agonist comprising or consisting of the sequence: X 1 is H or Y, X 2 is A or Aib, X 3 is D or E, X 7 is I, S, or T, X 11 is S or A, X 12 is T or I, X13 is I, Aib, or E; X 15 is D or E, X 16 is N, L, A, K, or E; X 17 is L, K, or I; X 20 is R or K, X 21 is D, E, or K; X 24 is N or E, X 28 is Q, A, or K, X 29 is T, Q, or K; X 30 is R or K, X 31 is I or G, X 32 is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29), or is omitted, However, X 7 If I, then X 2 is Aib and X 2 If A, then X 7 is S or T, the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of SEQ ID NO: 42; The peptide triple agonist, wherein the peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0284] 2. Arrays X 1 X 2 X 3 GTFX7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO: 43)-J A peptide triple agonist comprising or consisting of the sequence: X 1 is H or Y, X 2 But, Aib, X 3 is D or E, X 7 is I, S, or T, X 11 is S or A, X 12 is T or I, X 13 is I, Aib, or E; X 15 is D or E, X 16 is N, L, A, K, or E; X 17 is L, K, or I; X 20 is R or K, X 21 is D, E, or K; X 24 is N or E, X 28 is Q, A, or K, X 29 is T, Q, or K; X 30 is R or K, X 31 is I or G, X 32 is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29) or is omitted, the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of SEQ ID NO: 43; The peptide triple agonist, wherein the peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

[0285] 3. Arrays X 1 X 2 X 3 GTFX 7 SDYX 11 X 12 X 13 LX 15 X 16 X 17 AAX 20 X 21 FIX 24 WLIX 28 X 29 X 30 X 31 X 32 K (SEQ ID NO: 44)-J A peptide triple agonist comprising or consisting of the sequence: X 1 is H or Y, X 2 is A or Aib, X 3 is D or E, X 7 is S or T, X 11 is S or A, X 12 is T or I, X 13 is I, Aib, or E; X 15 is D or E, X 16 is N, L, A, K, or E; X 17 is L, K, or I; X 20 is R or K, X 21 is D, E, or K; X 24 is N or E, X 28 is Q, A, or K, X 29 is T, Q, or K; X 30 is R or K, X 31 is I or G, X 32 is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29) or is omitted, the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions in the C-terminal region of SEQ ID NO: 44; The peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor) and an agonist of GLP-2R (glucagon-like peptide-2 It is an agonist of the GLP-1R (glucagon-like peptide-1 receptor) and The peptide triple agonist is an agonist of the IL-1 receptor.

[0286] 4. The peptide triple agonist according to clause 1, wherein said C-terminal region of SEQ ID NO:42 consists of amino acid residues from position 16 to the C-terminus of SEQ ID NO:42.

[0287] 5. The peptide triple agonist according to any one of the preceding clauses, wherein said C-terminal region of SEQ ID NO:42 consists of amino acid residues 16 to 33 of SEQ ID NO:42.

[0288] 6. The peptide triple agonist according to any one of the preceding clauses, wherein said C-terminal region of SEQ ID NO:42 consists of amino acid residues 16 to 43 of SEQ ID NO:42.

[0289] 7. The peptide triple agonist of any one of the preceding clauses, wherein the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues at any one of positions 16 to 33 of SEQ ID NO: 42 and / or at any one of positions J (SEQ ID NO: 29).

[0290] 8.X 2 But, Aib, The peptide triple agonist of any one of the preceding clauses, wherein the peptide is modified at the C-terminus of SEQ ID NO:42 by the attachment of at least one fatty acid molecule.

[0291] 9.X 2 But, Aib, The peptide triple agonist of any one of the preceding clauses, wherein the peptide is modified by attaching at least one fatty acid molecule at the amino acid residue at position 33 of SEQ ID NO:42.

[0292] 10.X 16 is N, L, A, or E; 2. The peptide of any one of the preceding clauses, wherein the peptide is modified at the C-terminus of SEQ ID NO:42 by attachment of at least one fatty acid molecule.

[0293] 11.X 17 is L or I.

[0294] 12.X 20 is R.

[0295] 13.X 7The peptide trial agonist of any one of the preceding clauses, wherein

[0296] 14.X 7 The peptide trial agonist of any one of the preceding clauses, wherein

[0297] 15.X 13 The peptide trial agonist of any one of the preceding clauses, wherein is Aib.

[0298] 16.X 16 is N, A, or K.

[0299] 17.X 28 The peptide triple agonist of any one of the preceding clauses, wherein

[0300] 18.X 29 The peptide triple agonist of any one of the preceding clauses, wherein

[0301] 19.X 30 The peptide triple agonist of any one of the preceding clauses, wherein

[0302] 20.X 2 But Aib and X 31 But I and X 32 The peptide triple agonist of any one of the preceding clauses, wherein

[0303] 21.X 2 But Aib and X 31 But G and X 32 The peptide triple agonist of any one of the preceding clauses, wherein

[0304] 22.X 7 is T or S, and X 31 But G and X 32The peptide triple agonist of any one of the preceding clauses, wherein

[0305] 23.X 7 is T and X 31 But G and X 32 The peptide triple agonist of any one of the preceding clauses, wherein

[0306] 24. A peptide triple agonist according to any one of the preceding clauses, wherein J is omitted.

[0307] 25. The peptide triple agonist of any one of the preceding clauses, wherein J is selected from the group consisting of K, NDWKHNITQ, NDWKHNITQK.

[0308] 26. The peptide, H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 1), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 2); H-Aib-DGTFISDYATILDNLAARDFINWLIQTKITK (SEQ ID NO: 3), H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKITK (SEQ ID NO: 4), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK (SEQ ID NO: 5), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 6), H-Aib-DGTFISDYSTILENLAARDFINWLIQTKITK (SEQ ID NO: 7), H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKITK (SEQ ID NO: 8), H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKITK (SEQ ID NO: 9), H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKITK (SEQ ID NO: 10), H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 11), H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKITK (SEQ ID NO: 12), H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKITK (SEQ ID NO: 13), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGKK (SEQ ID NO: 14), H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKITK (SEQ ID NO: 15), Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 16), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 1), HADGTFTSDYSTILDNLAAKDFINWLIQTKGKK (SEQ ID NO: 17), HADGTFTSDYSTILDNLAARDFINWLIQTKGKK (SEQ ID NO: 18), HADGTFTSDYSIILDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 19), HADGTFTSDYSTILD-KLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 20), HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 21), HADGTFTSDYSTILDKKAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 22), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 23), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 24), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQK (SEQ ID NO: 25), HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQK (SEQ ID NO: 26), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK (SEQ ID NO: 27), HADGTFISDYSTILDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 45), HADGTFISDYST-Aib-LDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 46), HADGTFISDYSTILDNKAARDFINWLIQTKGKK-OH (SEQ ID NO: 47), HADGTFISDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 48), HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 49), and HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 50), or a functional variant thereof, wherein said functional variant comprises one or two individual amino acid substitutions.

[0309] 27. Is the peptide amidated at the C-terminus (-NH 2 ), or the C-terminus is a carboxylic acid.

[0310] 28. The peptide triple agonist of any one of the preceding clauses, wherein the peptide is modified by attaching at least one fatty acid molecule at a lysine (K) in any one of the positions in the C-terminal region of SEQ ID NO:42.

[0311] 29. The peptide triple agonist of any one of the preceding clauses, wherein the peptide is modified by the attachment of at least one fatty acid molecule at any one of positions 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, or 43 of SEQ ID NO:42.

[0312] 30. The peptide triple agonist of any one of the preceding clauses, wherein the peptide is modified by attaching at least one fatty acid molecule at any one of positions 16, 17, 20, 33, 34, 37, or 43 of SEQ ID NO:42.

[0313] 31. A peptide triple agonist according to any one of clauses 1, 9-11, wherein the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues at any one of positions 16, 17, 20-43 of SEQ ID NO: 42.

[0314] 32. The peptide triple agonist of any one of the preceding clauses, wherein the fatty acid molecule is a straight chain fatty acid.

[0315] 33. The peptide triple agonist of any one of the preceding clauses, wherein the fatty acid molecule is a branched fatty acid.

[0316] 34. The peptide triple agonist according to any one of the preceding clauses, wherein the fatty acid molecule is a monoacyl fatty acid molecule containing one fatty acid.

[0317] 35. The peptide triple agonist of any one of the preceding clauses, wherein the fatty acid molecule is a diacyl fatty acid molecule.

[0318] 36. The fatty acid molecule is CH 3 (CH 2 )8 CO-(Capryl, C10), CH 3 (CH 2 ) 10 CO-(Lauryl, C12), CH 3 (CH 2 ) 12 CO-(myristoyl, C14), CH 3 (CH 2 ) 14 CO-(palmitoyl, C16), CH 3 (CH 2 ) 16 CO-(stearyl, C18), and CH 3 (CH 2 ) 18 The peptide triple agonist of any one of the preceding clauses, comprising an acyl group selected from the group consisting of CO-(arachidyl, C20).

[0319] 37.The fatty acid molecule is HOOC-CH 3 (CH 2 ) 8 CO-(decanoyl, C10), HOOC-CH 3 (CH 2 ) 10 CO-(dodecanoyl, C12), HOOC-CH 3 (CH 2 ) 12 CO-(1-tetradecanoyl, C14), HOOC-CH 3 (CH 2 ) 14 CO-(hexadecanoyl, C16), HOOC-CH 3 (CH 2 ) 15 CO-(15-carboxy-pentadecanoyl, C17), HOOC-CH 3 (CH 2 ) 16 CO-(octadecanoyl, C18), HOOC-CH 3 (CH 2 ) 17 CO-(17-carboxy-heptadecanoyl, C19), HOOC-CH 3 (CH 2 ) 18 CO-(eicosanoyl, C20) and HOOC-CH3 (CH 2 ) 19 The peptide triple agonist of any one of the preceding clauses, comprising two acyl groups individually selected from the group consisting of CO-(19-carboxy-nonadecanoyl, C21).

[0320] 38. The peptide triple agonist according to any one of the preceding clauses, wherein the fatty acid molecule is attached to the amino acid residue via a spacer.

[0321] 39. The peptide triple agonist of any one of the preceding clauses, wherein the spacer comprises yGlu.

[0322] 40. The peptide triple agonist according to any one of the preceding clauses, wherein the spacer comprises or consists of yGlu or yGlu-yGlu.

[0323] 41. The peptide, a. an efficacy (E) that is at least 65% of the efficacy of native human GIP to activate human GIPR, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GIP to activate human GIPR max value) to activate human GIPR, b. an efficacy (E) that is at least 65% of the efficacy of native human GLP-2 to activate human GLP-2R, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GLP-2 to activate human GLP-2R max value) and activates human GLP-2R. c. an efficacy (E) that is at least 65% of the efficacy of native human GLP-1 to activate human GLP-1R, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GLP-1 to activate human GLP-1R max value) and activates human GLP-1R. A peptide triple agonist according to any one of the preceding clauses.

[0324] 42. The peptide, a EC of 10 nM or less against human GIPR 50 having b. EC of 50 nM or less for human GLP-2R 50 and c. EC value for human GLP-1R of 12 nM or less 50 having A peptide triple agonist according to any one of the preceding clauses.

[0325] 43. The peptide, H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 1), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-OH (SEQ ID NO:2); H-Aib-DGTFISDYATILDNLAARDFINWLIQTKIT-K(C10)-OH (SEQ ID NO: 3), H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 4), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO:5), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 6), H-Aib-DGTFISDYSTILENLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 7), H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 8), H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 9), H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 10), H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 11), H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 12), H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 13), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGK-K(C10)-NH 2 (SEQ ID NO: 14), H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 15), Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH 2 (SEQ ID NO: 16), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(γGlu-C10)-NH 2 (SEQ ID NO: 1), HADGTFTSDYSTILDNLAA-K(2×AEEA-γGlu-C18-diacid)-DFINWLIQTKGKK-OH (SEQ ID NO: 17), HADGTFTSDYSTILDNLAARDFINWLIQTKGK-K(γGlu-C18-diacid)-OH (SEQ ID NO: 18), HADGTFTSDYSIILD-K(2×AEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 19), HADGTFTSDYSTILD-K(2×AEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 20), HADGTFTSDYST-Aib-LD-K(2xAEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 21), HADGTFTSDYSTILDK-K (2xAEEA-γGlu-C18-diacid)-AARDFINWLIQTKGKKNDWKHNITQ-OH SEQ ID NO: 22), HADGTFTSDYSTILDKLAARDFINWLIQTKGK-K(2×AEEA-γGlu-C18-diacid)-NDWKHNITQ-OH (SEQ ID NO: 23), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDW-K(2×AEEA-γGlu-C18-diacid)-HNITQ-OH (SEQ ID NO: 24), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-K(2×AEEA-γGlu-C18-diacid)-OH (SEQ ID NO: 25), HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ-K (2×AEEA-γGlu-C18-diacid)-OH SEQ ID NO: 26), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKIT-K(2×AEEA-γGlu-C18-diacid)-OH (SEQ ID NO: 27), HADGTFISDYSTILDK-(2xAEEA-yGlu-C18-diacid)-LAARDFINWLIQTKGKK-OH (SEQ ID NO: 45), HADGTFISDYST-Aib-LDK-(2xAEEA-yGlu-C18-diacid)-LAARDFINWLIQTKGKK-OH (SEQ ID NO: 46), HADGTFISDYSTILDNK-(2xAEEA-yGlu-C18-diacid)-AARDFINWLIQTKGKK-OH (SEQ ID NO: 47), HADGTFISDYSTILDNLAARDFINWLIQTKGKK-(2×AEEA-yGlu-C18-diacid)-OH (SEQ ID NO: 48), HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-(2×AEEA-yGlu-C18-diacid)-OH (SEQ ID NO: 49), and HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-(2×AEEA-yGlu-C18-diacid)-OH (SEQ ID NO: 50), or a functional variant thereof, wherein said functional variant comprises one or two individual amino acid substitutions.

[0326] 44. A peptide triple agonist according to any of the preceding clauses for use as a medicament.

[0327] 45. A peptide triple agonist according to any one of the preceding clauses for use in a method for inhibiting bone resorption and / or stimulating bone formation.

[0328] 46. ​​A peptide triple agonist according to any one of the preceding clauses for use in a method for treating a bone disorder.

[0329] 47. The peptide triple agonist for use according to any one of the preceding clauses, wherein said bone disorder is selected from the group consisting of osteopenia, osteoporosis, severe osteoporosis, postmenopausal osteoporosis, idiopathic osteoporosis, osteomalacia, rickets, osteitis fibrosa cystic (OFC), and Paget's disease of bone.

[0330] 48. A peptide triple agonist according to any one of the preceding clauses for use in a method for treating idiopathic osteoporosis.

[0331] 49. A peptide triple agonist according to any one of the preceding clauses for use in a method for treating osteoporosis in an individual suffering from Duchenne muscular dystrophy (DMD) or cerebral palsy.

Claims

1. array X １ X ２ X ３ GTFX ７ SDYX １１ X １２ X １３ LX １５ X １６ X １７ AAX ２０ X ２１ FIX ２４ WLIX ２８ X ２９ X ３０ X ３１ X ３２ K (SEQ ID NO: 42) - J A peptide triple agonist comprising or consisting of the sequence: X １ is H or Y; X ２ is A or Aib, X ３ is D or E, X ７ is I, S, or T; X １１ is S or A, X １２ is T or I, X １３ is I, A, or E; X １５ is D or E, X １６ is N, L, A, K, or E; X １７ is L, K, or I; X ２０ is R or K, X ２１ is D, E, or K; X ２４ is N or E, X ２８ is Q, A, or K; X ２９ is T, Q, or K; X ３０ is R or K, X ３１ is I or G, X ３２ is T or K, J is a peptide comprising one or more amino acid residues of the sequence NDWKHNITQK (SEQ ID NO:29), or is omitted, However, X ７ When is I, X ２ is Aib, and X ２ If A, then X ７ is S or T, The peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues in any one of the positions of the C-terminal region of SEQ ID NO: 42, the C-terminal region of SEQ ID NO: 42 consisting of amino acid residues from position 16 to the C-terminus of SEQ ID NO: 42; The peptide triple agonist, wherein the peptide is an agonist of GIPR (glucose-dependent insulinotropic polypeptide receptor), an agonist of GLP-2R (glucagon-like peptide-2 receptor), and an agonist of GLP-1R (glucagon-like peptide-1 receptor).

2. 2. A peptide triple agonist according to any one of the preceding claims, wherein said C-terminal region of SEQ ID NO:42 consists of amino acid residues 16 to 33 of SEQ ID NO:

42.

3. 2. A peptide triple agonist according to any one of the preceding claims, wherein said C-terminal region of SEQ ID NO:42 consists of amino acid residues 16 to 43 of SEQ ID NO:

42.

4. 2. A peptide triple agonist according to any one of the preceding claims, wherein the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues at any one of positions 16 to 33 of SEQ ID NO:42 and / or at any one of positions J (SEQ ID NO:29).

5. X ２ is Aib, 2. A peptide triple agonist according to any one of the preceding claims, wherein the peptide is modified at the C-terminus of SEQ ID NO:42 by attachment of at least one fatty acid molecule.

6. X ２ is Aib, 10. A peptide triple agonist according to any one of the preceding claims, wherein the peptide is modified by attaching at least one fatty acid molecule at the amino acid residue at position 33 of SEQ ID NO:

42.

7. X １６ is N, L, A, or E; 2. A peptide according to any one of the preceding claims, wherein the peptide is modified at the C-terminus of SEQ ID NO:42 by attachment of at least one fatty acid molecule.

8. X １７ A peptide according to any one of the preceding claims, wherein is L or I.

9. X ２０ A peptide according to any one of the preceding claims, wherein

10. X ７ A peptide trial agonist according to any one of the preceding claims, wherein:

11. X ７ A peptide trial agonist according to any one of the preceding claims, wherein:

12. X １３ 4. The peptide trial agonist of any one of the preceding claims, wherein is Aib.

13. X １６ A peptide triple agonist according to any one of the preceding claims, wherein is N, A or K.

14. X ２８ A peptide triple agonist according to any one of the preceding claims, wherein:

15. X ２９ A peptide triple agonist according to any one of the preceding claims, wherein:

16. X ３０ A peptide triple agonist according to any one of the preceding claims, wherein

17. X ２ is Aib, and X ３１ is I and X ３２ A peptide triple agonist according to any one of the preceding claims, wherein:

18. X ２ is Aib, and X ３１ is G and X ３２ A peptide triple agonist according to any one of the preceding claims, wherein

19. X ７ is T or S, and X ３１ is G and X ３２ A peptide triple agonist according to any one of the preceding claims, wherein

20. X ７ is T and X ３１ is G and X ３２ A peptide triple agonist according to any one of the preceding claims, wherein

21. 2. A peptide triple agonist according to any one of the preceding claims, wherein J is omitted.

22. 10. A peptide triple agonist according to any one of the preceding claims, wherein J is selected from the group consisting of K, NDWKHNITQ, NDWKHNITQK.

23. The peptide is H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 1), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 2), H-Aib-DGTFISDYATILDNLAARDFINWLIQTKITK (SEQ ID NO: 3), H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKITK (SEQ ID NO: 4), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK (SEQ ID NO:5), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 6), H-Aib-DGTFISDYSTILENLAARDFINWLIQTKITK (SEQ ID NO: 7), H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKITK (SEQ ID NO: 8), H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKITK (SEQ ID NO: 9), H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKITK (SEQ ID NO: 10), H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 11), H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKITK (SEQ ID NO: 12), H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKITK (SEQ ID NO: 13), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGKK (SEQ ID NO: 14), H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKITK (SEQ ID NO: 15), Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 16), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKITK (SEQ ID NO: 1), HADGTFTSDYSTILDNLAAKDFINWLIQTKGKK (SEQ ID NO: 17), HADGTFTSDYSTILDNLAARDFINWLIQTKGKK (SEQ ID NO: 18), HADGTFTSDYSIILDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 19), HADGTFTSDYSTILD-KLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 20), HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 21), HADGTFTSDYSTILDKKAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 22), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 23), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ (SEQ ID NO: 24), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQK (SEQ ID NO: 25), HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQK (SEQ ID NO: 26), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKITK (SEQ ID NO: 27), HADGTFISDYSTILDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 45), HADGTFISDYST-Aib-LDKLAARDFINWLIQTKGKK-OH (SEQ ID NO: 46), HADGTFISDYSTILDNKAARDFINWLIQTKGKK-OH (SEQ ID NO: 47), HADGTFISDYSTILDNLAARDFINWLIQTKGKK-OH (SEQ ID NO: 48), HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 49), and HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-OH (SEQ ID NO: 50), 5. A peptide triple agonist according to any one of the preceding claims, selected from the group consisting of:

24. The peptide is either amidated (-NH ２ ), or the C-terminus is a carboxylic acid.

25. A peptide triple agonist according to any one of the preceding claims, wherein the peptide is modified by attaching at least one fatty acid molecule to a lysine (K) at any one of the positions in the C-terminal region of SEQ ID NO:

42.

26. 2. A peptide triple agonist according to any one of the preceding claims, wherein the peptide is modified by the attachment of at least one fatty acid molecule at any one of positions 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42 or 43 of SEQ ID NO:

42.

27. A peptide triple agonist according to any one of the preceding claims, wherein the peptide is modified by attaching at least one fatty acid molecule at any one of positions 16, 17, 20, 33, 34, 37, or 43 of SEQ ID NO:

42.

28. A peptide triple agonist according to any one of claims 1, 9 to 11, wherein the peptide is modified by attaching at least one fatty acid molecule at one or more amino acid residues at any one of positions 16, 17, 20 to 43 of SEQ ID NO:

42.

29. 10. A peptide triple agonist according to any one of the preceding claims, wherein the fatty acid molecule is a straight chain fatty acid.

30. 10. A peptide triple agonist according to any one of the preceding claims, wherein the fatty acid molecule is a branched fatty acid.

31. 10. A peptide triple agonist according to any one of the preceding claims, wherein the fatty acid molecule is a monoacyl fatty acid molecule containing one fatty acid.

32. 10. A peptide triple agonist according to any one of the preceding claims, wherein the fatty acid molecule is a diacyl fatty acid molecule.

33. The fatty acid molecule is CH ３ (CH ２ ) ８ CO- (Capryl, C10), CH ３ (CH ２ ) １０ CO-(Lauryl, C12), CH ３ (CH ２ ) １２ CO-(myristoyl, C14), CH ３ (CH ２ ) １４ CO-(palmitoyl, C16), CH ３ (CH ２ ) １６ CO- (stearyl, C18), and CH ３ (CH ２ ) １８ 2. A peptide triple agonist according to any one of the preceding claims, comprising an acyl group selected from the group consisting of: CO-(arachidyl, C20).

34. The fatty acid molecule is HOOC-CH ３ (CH ２ ) ８ CO-(decanoyl, C10), HOOC-CH ３ (CH ２ ) １０ CO-(dodecanoyl, C12), HOOC-CH ３ (CH ２ ) １２ CO-(1-tetradecanoyl, C14), HOOC-CH ３ (CH ２ ) １４ CO-(hexadecanoyl, C16), HOOC-CH ３ (CH ２ ) １５ CO-(15-carboxy-pentadecanoyl, C17), HOOC-CH ３ (CH ２ ) １６ CO-(octadecanoyl, C18), HOOC-CH ３ (CH ２ ) １７ CO-(17-carboxy-heptadecanoyl, C19), HOOC-CH ３ (CH ２ ) １８ CO- (eicosanoyl, C20), and HOOC-CH ３ (CH ２ ) １９ 2. A peptide triple agonist according to any one of the preceding claims, comprising two acyl groups individually selected from the group consisting of: CO-(19-carboxy-nonadecanoyl, C21).

35. 2. A peptide triple agonist according to any one of the preceding claims, wherein the fatty acid molecule is attached to the amino acid residue via a spacer.

36. 2. A peptide triple agonist according to any one of the preceding claims, wherein the spacer comprises yGlu.

37. 10. A peptide triple agonist according to any one of the preceding claims, wherein the spacer comprises or consists of yGlu or yGlu-yGlu.

38. The peptide is a. an efficacy (E) that is at least 65% of the efficacy of native human GIP to activate human GIPR, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GIP to activate human GIPR; ｍａｘ value) to activate human GIPR; b. an efficacy (E) that is at least 65% of the efficacy of native human GLP-2 to activate human GLP-2R, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GLP-2 to activate human GLP-2R. ｍａｘ value) to activate human GLP-2R; c. an efficacy (E) that is at least 65% of the efficacy of native human GLP-1 to activate human GLP-1R, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95% of the efficacy of native human GLP-1 to activate human GLP-1R. ｍａｘ value) to activate human GLP-1R; 10. A peptide triple agonist according to any one of the preceding claims.

39. The peptide is d. EC50 of 10 nM or less against human GIPR ５０ having e. EC50 nM or less for human GLP-2R ５０ and f. EC50 of 12 nM or less for human GLP-1R ５０ having 10. A peptide triple agonist according to any one of the preceding claims.

40. The peptide is H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO:1), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-OH (SEQ ID NO: 2), H-Aib-DGTFISDYATILDNLAARDFINWLIQTKIT-K(C10)-OH (SEQ ID NO: 3), H-Aib-DGTFISDYSTILDLLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO:4), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO:5), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO:6), H-Aib-DGTFISDYSTILENLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO: 7), H-Aib-DGTFISDYSTILDNLAAREFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO:8), H-Aib-DGTFISDYSTILDNLAARDFIEWLIQTKIT-K(C10)-NH ２ (SEQ ID NO: 9), H-Aib-DGTFISDYSIILDNLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO: 10), H-Aib-EGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO:11), H-Aib-DGTFISDYST-Aib-LDNLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO: 12), H-Aib-DGTFISDYSTELDNLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO: 13), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKGK-K(C10)-NH ２ (SEQ ID NO: 14), H-Aib-DGTFISDYSTILDNIAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO: 15), Y-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K(C10)-NH ２ (SEQ ID NO: 16), H-Aib-DGTFISDYSTILDNLAARDFINWLIQTKIT-K (γGlu-C10)-NH ２ (SEQ ID NO:1), HADGTFTSDYSTILDNLAA-K(2xAEEA-γGlu-C18-diacid)-DFINWLIQTKGKK-OH (SEQ ID NO: 17), HADGTFTSDYSTILDNLAARDFINWLIQTKGK-K(γGlu-C18-diacid)-OH (SEQ ID NO: 18), HADGTFTSDYSIILD-K(2xAEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO: 19), HADGTFTSDYSTILD-K(2xAEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO:20); HADGTFTSDYST-Aib-LD-K(2xAEEA-γGlu-C18-diacid)-LAARDFINWLIQTKGKKNDWKHNITQ-OH (SEQ ID NO:21), HADGTFTSDYSTILDK-K (2xAEEA-γGlu-C18-diacid)-AARDFINWLIQTKGKKNDWKHNITQ-OH SEQ ID NO:22), HADGTFTSDYSTILDKLAARDFINWLIQTKGK-K(2xAEEA-γGlu-C18-diacid)-NDWKHNITQ-OH (SEQ ID NO:23), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDW-K(2xAEEA-γGlu-C18-diacid)-HNITQ-OH (SEQ ID NO:24), HADGTFTSDYSTILDKLAARDFINWLIQTKGKKNDWKHNITQ-K(2xAEEA-γGlu-C18-diacid)-OH (SEQ ID NO:25), HADGTFTSDYST-Aib-LDKLAARDFINWLIQTKGKKNDWKHNITQ-K (2xAEEA-γGlu-C18-diacid)-OH SEQ ID NO: 26), H-Aib-DGTFISDYSTILDALAARDFINWLIQTKIT-K(2xAEEA-γGlu-C18-diacid)-OH (SEQ ID NO:27), HADGTFISDYSTILDK-(2xAEEA-yGlu-C18-diacid)-LAARDFINWLIQTKGKK-OH (SEQ ID NO:45), HADGTFISDYST-Aib-LDK-(2xAEEA-yGlu-C18-diacid)-LAARDFINWLIQTKGKK-OH (SEQ ID NO:46); HADGTFISDYSTILDNK-(2xAEEA-yGlu-C18-diacid)-AARDFINWLIQTKGKK-OH (SEQ ID NO:47); HADGTFISDYSTILDNLAARDFINWLIQTKGKK-(2xAEEA-yGlu-C18-diacid)-OH (SEQ ID NO:48), HADGTFISDYSTILDNLAARDFINWLIQTKGKKK-(2xAEEA-yGlu-C18-diacid)-OH (SEQ ID NO:49), and HADGTFISDYST-Aib-LDNLAARDFINWLIQTKGKKK-(2xAEEA-yGlu-C18-diacid)-OH (SEQ ID NO:50), 5. A peptide triple agonist according to any one of the preceding claims, selected from the group consisting of:

41. A peptide triple agonist according to any of the preceding claims for use as a medicament.

42. 13. A peptide triple agonist according to any one of the preceding claims for use in a method for inhibiting bone resorption and / or stimulating bone formation.

43. 10. A peptide triple agonist according to any one of the preceding claims for use in a method for treating a bone disorder.

44. 2. The peptide triple agonist for use according to any one of the preceding claims, wherein said bone disorder is selected from the group consisting of osteopenia, osteoporosis, severe osteoporosis, postmenopausal osteoporosis, idiopathic osteoporosis, osteomalacia, rickets, osteitis fibrosa cystic (OFC) and Paget's disease of bone.

45. 10. A peptide triple agonist according to any one of the preceding claims for use in a method for treating idiopathic osteoporosis.

46. 10. A peptide triple agonist according to any one of the preceding claims for use in a method for treating osteoporosis in an individual suffering from Duchenne muscular dystrophy (DMD) or cerebral palsy.

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