Peptide targeted nuclear hormones

Abstract

Provided herein are incretin peptides conjugated with NHR ligands, wherein the conjugates are active at a nuclear hormone receptor. Also provided herein are pharmaceutical compositions and kits of the conjugates of the invention. Further provided herein are methods of treating a disease, e.g., a metabolic disorder, such as diabetes and obesity, comprising administering the conjugates of the invention.

Description

32993-439275PEPTIDE TARGETED NUCLEAR HORMONESCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U. S. Provisional Patent Application No. 63 / 791,104 filed on April 18, 2025, the disclosure of which is expressly incorporated herein.INCORPORATION BY REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY

[0002] Incorporated by reference in its entirety is a computer-readable nucleotide / amino acid sequence listing submitted concurrently herewith and identified as follows: 114,000 bytes xml file named ‘’42O454.xml” created on April 15, 2026.BACKGROUND

[0003] Nuclear hormone receptor proteins form a class of ligand activated proteins that, when bound to specific sequences of DNA, serve as on-off switches for transcription within the cell nucleus. These switches control the development and differentiation of skin, bone and behavioral centers in the brain, as well as the continual regulation of reproductive tissues.

[0004] The interaction of nuclear hormone and receptor triggers a conformational change in the receptor, which results in induction of gene expression. The level of cellular signal transduction activated by the interaction of a ligand and a nuclear hormone receptor is determined by the number of ligands and receptors available for binding, and by the binding affinity between the ligand and the receptor. Many ligands and corresponding analogs that bind to nuclear hormone receptors are used as medication to treat, for example, Parkinson’s disease (NURR1), sleep disorders (RZR0), arthritis and cerebellar ataxia (RORa), central nervous system disorders (NOR-1, Rev-Erb A0. Tlx, NGFI-B0, HZF-2a, COUP-TFa, COUP-TF0, COUR-TFy, NUR77), hypercholesterolemia (LXRa, COR), obesity (Rev-ErbAa), diabetes (HNF4a), immune disorders (TOR), metabolic disorders (MB67a, SHP, FXR, SF-1, LXR0), and infertility and contraception (GCNF, TR2-I la.ty TR4, ERa,0 ERRa,0).

[0005] Pre-proglucagon is a 158 amino acid precursor polypeptide that is processed in different tissues to form a number of different proglucagon-derived peptides, including glucagon, glucagon-like peptide-1 (GLP-1), glucagon-hke peptide-2 (GLP-2) and oxyntomodulin (OXM), that are involved in a wide variety of physiological functions, including32993-439275glucose homeostasis, insulin secretion, gastric emptying, and intestinal growth, as w ell as the regulation of food intake. Glucagon is a 29-amino acid peptide that corresponds to amino acids 33 through 61 of pre-proglucagon, while GLP-1 is produced as a 37-amino acid peptide that corresponds to amino acids 72 through 108 of pre-proglucagon. GLP-l(7-36) amide or GLP-1(7-37) acid are biologically potent forms of GLP-1, that demonstrate essentially equivalent activity at the GLP-1 receptor.

[0006] Glucagon is used in the acute treatment of severe hypoglycemia.Oxyntomodulin has been reported to have pharmacological ability7to suppress appetite and lower body weight. GLP-1 and GLP-1 receptor agonists are used as treatment for Type II diabetes. Exendin-4 is a peptide present in the saliva of the Gila monster that resembles GLP-1 in structure, and like glucagon and GLP-1, increases insulin biosynthesis and secretion.

[0007] Gastric inhibitory polypeptide (GIP) is also known as a glucose-dependent insulinotropic peptide, and is a member of the secretin family of hormones. GIP is derived from a 153-amino acid proprotein encoded by the GIP gene and circulates principally as a biologically active 42-amino acid peptide. The GIP gene is expressed in the small intestine as well as the salivary7glands and was first characterized as an inhibitor of gastric acid secretion. In addition to its inhibitory7effects in the stomach, in the presence of glucose, GIP enhances insulin synthesis and secretion by pancreatic beta islet cells when administered in physiological doses. GIP is an enteric factor that physiologically stimulates the release of pancreatic insulin and serves a role in maintaining glucose homeostasis.

[0008] GIP and GLP-1 are known members of the incretin peptide family, and incretin receptor signaling is known to exert physiologically relevant actions that are critical for glucose homeostasis. In normal physiology, GIP and GLP-1 are secreted from the gut following a meal, and these incretins enhance the physiological response to food including sensation of satiety, insulin secretion, and nutrient disposal. T2D patients have impaired incretin responses.

[0009] Dosing diabetes patients with GLP-1 analogs has been found to be limited by adverse effects, such as nausea and vomiting, and consequently dosing most often cannot reach full efficacy for glycemic control and weight loss. GIP alone has very modest glucose-lowering ability in type 2 diabetic humans. Both native GIP and GLP-1 are inactivated rapidly by the ubiquitous protease, DPP IV, and therefore, can only be used for short-term metabolic control.

[0010] Peptides that constitute balanced, full potency agonists at the GLP-1, glucagon (Gcg) and GIP receptors have demonstrated transformative pharmacology7in the treatment of metabolic diseases, and most notably reduction in body weight. The most effective drug32993-439275(tirzepatide, Mounjaro / Zepbound) is formulated as a subcutaneous injection and administered weekly. This peptide requires a scheduled dose titration to minimize adverse gastrointestinal effects, with monthly increments over the course of a five-month period.

[0011] There remains a need for peptides possessing increased efficacy and less propensity for adverse effects to treat excess body weight and the associated diseases that lessen the quality and quality of life in adults. The present disclosure is directed to novel conjugates of nuclear hormone receptor ligands and incretins.SUMMARY OF THE INVENTION

[0012] Provided herein are conjugate entities comprising an incretin peptide covalently linked to a nuclear hormone receptor ligand (an ‘NHR ligand”). These conjugates have plural activities and are useful for the treatment of a variety of diseases.

[0013] In one embodiment a compound comprising the structure Q-L-Y is provided wherein;Q is an incretin peptide;Y is a NHR ligand, optionally a peroxisome proliferator activated receptor (PPAR) agonist; andL is a bond or a linking group covalently linking Q to Y, optionally with the proviso that Q is not lanifibranor. In one embodiment Y is linked to the side chain of an amino acid located in the C-terminal region of the incretin peptide, optionally at a position downstream from position 20. including for example at position 24, 31, 40 or at an amino acid of a C-terminal extension of the incretin peptide. In one embodiment Q is an incretin peptide wherein the C-terminal amino acid is amidated and Y is linked to the C-terminal amide of the incretin peptide. In one embodiment L is a bond and Q and Y are directly linked via an amide bond. In one embodiment L is a linking group selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-8 and q and z are independently an integer selected from the range of 0-4. In one embodiment L is a linker comprising the structure -[COCH2(OCH2CH2)k-NH]q, wherein k is an integer from 2-8 and q is 1.

[0014] In one embodiment Q is an incretin peptide selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-132993-439275co-agonist, and a glucagon / GIP / GLP-1 tri-agonist, and Y is linked to Q via the side chain of an amino acid selected from positions 12, 17, 20, 24, 28, 30 and any amino acid C-terminal to position 30, including any amino acid added to the C-terminus of the incretin peptide. In one embodiment Y is linked to the amino acid side chain at position 40 of the incretin peptide. In one embodiment Y is a PPAR agonist selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar and related analogs of those compounds that retain activity as PPAR agonists, and in one embodiment Y is GW501516.

[0015] In one embodiment Q is an incretin peptide selected from peptides comprising the sequencei) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSX40(SEQ ID NO: 19); orii) Y-Aib-QGTFTSDYSIXi3LDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSX4o (SEQ ID NO: 20), oriii) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPX4o (SEQ ID NO: 21); oriv) H-Aib-EGTFTSDVSSYLEGQAAKEFlAWLVRGRGX4o (SEQ ID NO: 22); orv) H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;X13 is alpha-methyl leucine;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys, Om, Cys, or the amino acid at X40 is amidated and linked to -[COCH2(OCH2CH2)k-NH]qwherein k is an integer selected from 1-8 and q is 1 or 2, whereina NHR ligand is linked to the side chain of an amino acid at position 12, 24 or 40 of said incretin peptide, or at a C-terminal primary amine of said incretin peptide, optionally wherein the NHR ligand is linked to a side chain of an amino acid of the incretin peptide via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acidgamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P. wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and z are independently an integer selected from the range of 0-4.32993-439275

[0016] In accordance with one embodiment of the present disclosure, a dimer or multimer of an incretin peptide / NHR ligand conjugate is provided wherein a first and second incretin peptide are linked together via a bridging linker and a nuclear hormone receptor ligand is linked to one or both of the first and second incretin peptides. In one embodiment, the first and second incretin peptides are independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 co-agonist, and a Gcg / GIP / GLP-1 tri-agonist. In one embodiment the first and second incretin peptides are independently selected from the group consisting of a GIP / GLP-1 co-agonist, and a Gcg / GIP / GLP-1 tri-agonist. In one embodiment the first and second incretin peptides are linked together via a bridging-linker joiningi) the side chain of an amino acid independently selected from positions 12, 17, 24, 31 and 40 of the respective first and second incretin peptide, optionally at position 12 or 24, orii) a C-terminal amide of the first incretin peptide to a C-terminal amide of the second incretin peptide; oriii) a C-terminal amide of the first incretin peptide to the side chain of an amino acid selected from positions 12, 24, 31 and 40 of the second incretin peptide.

[0017] In one embodiment the NHR ligand of the incretin peptide / NHR ligand conjugate is linked to the side chain of an amino acid of the first and / or second incretin peptide at a position not linked to the bridging linker, and independently selected from position 24, 31 or 40, optionally at position 12 or 24. In one embodiment the NHR ligand is linked to the side chain of an amino acid at position 40 of the first and / or second incretin peptide.

[0018] In one embodiment a conjugate comprising a dimer of an incretin peptide and an NHR ligand is provided wherein a first and second incretin peptide are linked to one another to form an incretin peptide heterodimer or homodimer, and the NHR ligand is linked to the first and / or second incretin peptide. In one embodiment the first and second incretin peptides of the incretin peptide heterodimer or homodimer are independently selected from peptides comprising the sequencei) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSX4o (SEQ ID NO: 19); oru) Y-Aib-QGTFTSDYSIXi3LDKKAQ-Aib-AFlEYLLEGGPSSGAPPPSX4o (SEQ ID NO: 20), or32993-439275iii) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPXw (SEQ ID NO: 21); oriv) H-Aib-EGTFTSDVSSYLEGQAAKEFIAWLVRGRGX4o (SEQ ID NO: 22); orv) H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;X13 is alpha-methyl leucine;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys, Om, Cys, or the amino acid at X40 is amidated and linked to -tCOCH2(OCH2CH2)k-NH]qwherein k is an integer selected from 1-8 and q is 1 or 2, wherein the first and second incretin peptides are linked together via a bridging linker joining the side chains of amino acids independently selected from position 12, 24, 31 or 40 of the respective first and second incretin, anda nuclear hormone receptor ligand is linked to the side chain of an amino acid at a position not linked to the bridging linker and independently located at position 12, 24 or 40 of said first and / or second incretin peptide, or at a C-terminal primary7amine of said first and / or second incretin peptide, optionally wherein the nuclear hormone receptor ligand is linked to a side chain of an amino acid of the first and / or second incretin peptide via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4. In one embodiment the bridging linker comprises a structure selected from:i) a disulfide bond;ii) -NH(CO)(CH2)n-S-, wherein n is 1-4;iii) a triazole; oriv) -[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]s wherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2.

[0019] In a further embodiment the nuclear hormone receptor ligand is directly linked (i.e., without a spacer) to the side chain of the Lys located at position 40 of said first and / or second incretin peptide and the bridging linker links the side of amino acids located at position 12 of the first incretin peptide to position 12 of the second incretin peptide, or the bridging32993-439275linker links the side of amino acids located at position 24 of the first incretin peptide to position 24 of the second incretin peptide.

[0020] In one embodiment the NHR ligand linked to the first and / or second incretin peptide is independently selected from a PPAR agonist, a famesoid X receptor (FXR) agonist such as cilofexor, or a steroid, optionally wherein the PPAR agonist is a compound selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, and the steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol.

[0021] In one embodiment a conjugate is provided comprising a multimer of covalently linked incretin peptides, optionally wherein the multimer comprises a dimer of an incretin peptide, and an NHR ligand covalently linked to at least one of the incretin peptides of the multimer. In one embodiment the multimer comprises a first and second incretin peptide linked to one another to form an insulin peptide heterodimer or homodimer wherein the first incretin peptide comprises the sequence ofH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys; andthe second incretin peptide comprising the sequenceH-Aib-EGTFTSDVSXi02YLEXi06QAAKEFIX204WLVX208GRPSSGAPPPSX400 (SEQ ID NO: 24), whereinX102 is Ser, Lys or Cys;X106 is Gly or Glu;X204 is Ala, Lys or Cys;X208 is Arg or Lys; andX400 is Lys, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chains of the Lys at position 40 of the respective first and second incretin peptides, said bridging linker comprising the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2; further wherein32993-439275a nuclear hormone receptor ligand is linked to the side chain of an amino acid of the first incretin peptide and / or the second incretin peptide at position independently selected from positions 12, 20, and 24, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4. In one embodiment the first and the second incretin peptides are liked to one another via any of the bridging linkers disclosed herein.

[0022] In one embodiment a conjugate is provided comprising an incretin peptide tetramer comprising any two of the incretin dimer / NHR conjugates disclosed herein, wherein the two incretin dimer / NHR conjugates are linked together via a tetramer linker. The tetramer linker can be any linker that provides spacing between the two incretin dimer / NHR conjugates. In one embodiment the tetramer comprises two sets of incretin peptide dimers wherein each incretin peptide dimer is covalently linked to an NHR ligand and the two incretin peptide dimers are covalently linked to one another via a tetramerizing linker. The incretin peptide tetramer can be formed by linking two incretin peptide homodimers to one another, or two incretin peptide heterodimers to one another or linking an incretin peptide heterodimer to an incretin homodimer. In one embodiment the two sets of incretin peptide dimers of the incretin peptide tetramer each comprise a bridging tetramer linker having the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]s wherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2, wherein said tetramerizing linker comprises the structure -(CH2)m(CHCONH2)-, wherein m is an integer selected from 1-8 that is linked to the primary amine of each of the respective bridging linkers of the two incretin peptide dimers comprising the incretin peptide tetramer, wherein a nuclear hormone receptor ligand is linked to the side chain of an amino acid of any one or more of the first, second, third or fourth incretin peptides comprising the incretin peptide tetramer at aposition independently selected from positions 12, 20, 24 or 28 of each incretin peptide, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acidjz-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acidjp, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.In one embodiment a conjugate is provided comprising an incretin peptide tetramer wherein four incretin / NHR ligand conjugates are linked to a PEG scaffold, wherein the tetramer has the general structure:32993-439275wherein n is an integer selected from about 100 to about 1000. or from about 200 to about 500, and grey rectangles represent an incretin peptide covalently bound to an NHR ligand. Each of the covalently linked incretin / NHR ligands of the tetramer can be the same, or different, and the four covalently linked incretin / NHR ligands can differ based on the incretin peptide (independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 co-agonist, and a glucagon / GIP / GLP-1 tri-agonist) and / or the NHR ligand. In one embodiment one or more of the four incretin peptides are acylated, and in an alternative embodiment none of the four incretin peptides are acylated. In one embodiment 1, 2, 3 or 4 individual incretin peptides (independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 co-agonist, and a glucagon / GIP / GLP-1 tri-agonist) are covalently linked to the PEG scaffold via the C-terminus of the incretin peptides using standard techniques, including click chemistry. In one embodiment, NHR ligands are covalently linked, optionally via a spacer, to 1.2, 3, or 4 of the incretin peptides of the tetramer at positions selected from 24, 20, 17, 16 and 12, optionally from positions 24, 20 or 17, optionally from positions 24 and 20, optionally at position 24. Any combination of incretin peptide and NHR ligand as disclosed herein can be linked to form the tetramers of the present disclosure. In one embodiment at least one of the incretin peptides is a GIP / GLP-1 co-agonist and at least on of the NHR ligands is GW501516.

[0023] In some aspects of the invention, prodrugs of the incretin peptide / nuclear hormone receptor ligand compounds are provided wherein the prodrug comprises a dipeptide prodrug element (A-B) covalently linked to the N-terminal alpha amine, or at the side chain of a Lys residue of one or more incretin peptides comprising the incretin peptide / nuclear hormone receptor ligand compound. In one embodiment the first amino acid of the dipeptide is acylated, optionally with a C16-C30 fatty acid, a C16-C30 phosphonic fatty acid, or a C16-C30 diacid.

[0024] In some aspects of the invention, pharmaceutical compositions comprising any of the incretin peptide / nuclear hormone receptor ligand compounds disclosed herein, and a pharmaceutically acceptable carrier are also provided.32993-439275

[0025] In other aspects of the invention, methods are provided for administering a therapeutically effective amount of an incretin peptide / nuclear hormone receptor ligand compound described herein for treating a disease or medical condition in a patient. In some embodiments, the disease or medical condition is selected from the group consisting of metabolic syndrome, diabetes, obesity, liver steatosis, and a neurodegenerative disease.BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Fig. 1 presents an alignment of the amino acid sequences of various glucagon superfamily peptides or relevant fragments thereof. The amino acid sequence presented are GHRH (SEQ ID NO: 7), PHM (SEQ ID NO: 10), VIP (SEQ ID NO: 8), PACAP-27 (SEQ ID NO: 9), Exendin-4 (SEQ ID NO: 6), GLP-1 (SEQ ID NO: 2), Glucagon (SEQ ID NO: 1), Oxyntomodulin (SEQ ID NO: 3), GIP (SEQ ID NO: 4), GLP-2 (SEQ ID NO: 5) and Secretin (SEQ ID NO: 11). The alignment shows how amino acid positions of glucagon can correspond to amino acid positions in other glucagon superfamily peptides.

[0027] Figs. 2A-2C provide data showing GLP-1 -mediated delivery of the PPARa / y dual-agonist Tesaglitazar (SEQ ID NO: 48) improves obesity and glucose metabolism in mice when administered as a daily subcutaneous dose of 50 nmol / kg (Quarta, et al (2022) Nature Metabolism 63:1071-83). Fig. 2A provides the structure of the GLP-1 agonist / Tesaglitazar conjugate (SEQ ID NO: 48); Fig. 2B is a graph showing weight loss over time for mice administered vehicle (o) GLP-1 agonist (•) Tesaglitazar (♦) or the GLP-1 agonist / Tesaglitazar conjugate (•). Fig. 2C shows cumulative food intake over 14 days in mice fed the various compounds.

[0028] Figs. 3A-3C provide data showing GLP-l / GIP co-agonist-mediated delivery7of the pan-PPAR-agonist lanifibinor as a daily subcutaneous dose at 10 nmol / kg improves obesity and glucose metabolism in mice. Fig. 3A provides the structure of the GLP-l / GIP coagonist / lanifibrinor conjugates tested, wherein MAR709 has the amino acid sequence of SEQ ID NO: 49; Fig. 3B is a graph showing weight loss over time for mice administered each day vehicle, BWB 2050, BWB 2051, BWB 2052, BWB 2053 or BWB 2050 at 10 nmol / kg. Fig. 3B shows the food intake over 14 days after administration of the compounds. The data shows impressive weight lowering (>15%) in two weeks at lOnmol / kg. Surprisingly, the distance of the lanifibranor from backbone of peptide is inversely related to efficacy^ (compare BWB 2051 with BWB 2052 and BWB 2053), with BWB 2051 providing weight loss 5x greater than the peptide w / o lanifibranor.32993-439275

[0029] Figs. 4A-4C provide data showing the results of GLP-l / GIP co-agonist-mediated delivery of various PPAR agonist compounds selected from Lanifibrinor, GW501516, Bezafibrate, Elafibranor and Pemafibrate to DOI mice as a single daily subcutaneous dose of lOnmol / kg. Fig. 4A provides the structure of the vanous GLP-l / GIP co-agonist / PPAR agonist conjugates tested; Fig. 4B is a graph showing weight loss over time for mice administered vehicle, BWB 2050, BWB 2051, BWB 2069, BWB 2070, BWB 2071, or BWB 2072 at 10 nmol / kg. The data shows the amount of weight lowering is dependent on the PPAR agonist linked to the GLP-l / GIP co-agonist. Fig. 4C a graph of the blood glucose levels (mg / dL) as a function of time before and after a glucose injection (administered at timepoint 0) in mice last injected with peptide 24 hours before the glucose injection (with a vehicle control, BWB 2050, BWB 2051, BWB 2069, BWB 2070, BWB 2071, or BWB 2072) at 10 nmol / kg. Surprisingly, BWB 2072 demonstrated superior glucose control although inducing less body weight loss relative to other conjugates.

[0030] Figs. 5A-5C provide data showing the results of subcutaneous injection of a GLP-l / GIP co-agonist-PPAR agonist conjugate verses a subcutaneous injection a GLP-1 agonist-PPAR agonist conjugate into DOI mice as a daily dose of 10 nmol / kg. Fig. 5A provides the structure of the BWB 1060 (SEQ ID NO: 50) and PPAR conjugates thereof: BWB 2081 and BWB 2082. Fig. 5B shows the change in body weight over time after administration of vehicle. BWB 1060, BWB 2081, or BWB 2082 to DOI mice. Fig. 5C shows the change in body weight over time after administration of vehicle, BWB 2050, BWB 2051, BWB 2069, or BWB 2071 to DOI mice.

[0031] Figs. 6A-6D provide data showing the in vitro activity of various glucagon agonist-PPAR agonist conjugates at the human glucagon receptor (hGR), the human GLP-1 receptor (hGLP-1) and the human GIP receptor (hGIP) as measured using cells transfected with the respective receptor genes. Fig. 6A shows the structure of the glucagon agonists (BWB 2092, BWB 2094, and BWB 2095) conjugated with the PPAR agonist GW501516 relative to native glucagon, native GLP-1 and native human GIP. Fig. 6B shows the activity of the compounds at the human glucagon receptor (hGR); Fig. 6C shows the activity of the compounds at the human GLP-1 receptor (hGLP-1) and Fig. 6D shows the activity of the compounds at the human GIP receptor (hGIP).

[0032] Figs. 7A-7B provide data showing the results of a single subcutaneous injection of a GLP-l / GIP co-agonist-PPAR agonist conjugate into DOI mice at a daily dose of 10 nmol / kg, wherein the GLP-l / GIP co-agonist is MAR709 and the PPAR agonist is selected from GW501516, chiglitazar and sarolitazar. Fig. 7A shows the structures of the conjugates32993-439275tested. Fig. 7B is a graph showing weight loss over time for mice administered vehicle, BWB 2050, BWB 2069, BWB 2081, BWB 2084 or BWB 2091 at 10 nmol / kg or 30 nmol / kg.

[0033] Fig. 8 provides a table showing the in vitro activity of GLP-l / CEX agonist dimers at the human GLP-1 receptor. The dimers are homodimers formed from GLP-l / CEX analogs comprising a substitution of Aib at position 2, Glu at position 16 and a CEX-extension sequence relative to the native GLP-1 sequence, that are linked to one another via a disulfide linked between cysteine substituted amino acids at position 12, 24, 31 or 40. All dimers were found to be active at the human GLP-1 receptor.

[0034] Figs. 9A-9G show the structure of various GLP-1 agonist dimers. Fig. 9A provides the structure of BWB 10080 (dimer formed between incretin peptides of SEQ ID NO: 46 and SEQ ID NO: 51); Fig. 9B provides the structure of BWB 10081 (dimer formed between incretin peptides of SEQ ID NO: 52 and SEQ ID NO: 53); Fig. 9C provides the structure of BWB 10084 (dimer formed between incretin peptides of SEQ ID NO: 46 and SEQ ID NO: 51, plus steroid); Fig. 9D provides the structure of BWB 10087 (dimer formed between incretin peptides of SEQ ID NO: 46 and SEQ ID NO: 51, plus two steroids); Fig. 9E provides the structure of BWB 10085 (dimer formed between incretin peptides of SEQ ID NO: 46 and SEQ ID NO: 51, plus a PPAR agonist); Fig. 9F provides the structure of BWB 10088 (dimer formed between incretin peptides of SEQ ID NO: 46 and SEQ ID NO: 51, plus two PPAR agonists); and Fig. 9G provides the structure of BWB 10086 (dimer formed between incretin peptides of SEQ ID NO: 46 and SEQ ID NO: 51, plus one steroid and one PPAR agonist). Fig. 9H provides the results of a daily subcutaneous injection of a GLP-1 agonist-NHR ligand monomer into DOI mice at a dose of 10 nmol / kg, wherein the NHR ligand is Estradiol (E2) or dexamethasone (Dexa) relative to the parent GLP-1 agonist dimer.

[0035] Figs. 10A-10C provide the in vitro activity of a GLP-1 agonist dimer and GLP-1 agonist tetramer at at the human glucagon receptor (hGR), the human GLP-1 receptor (hGLP-1) and the human GIP receptor (hGIP) as measured using cells transfected with the respective receptor genes. Fig. 10A shows the structure of dimer 2x(MAR709-2xOEG)dK (BWB 2106) and Fig. 10B shows the structure of tetramer 2x(2x(MAR709-2xOEG)dK)dK (BWB 2107). Fig. 10C provides the relative activity of Mar709 (BMB 2050), BMB 2106, BMB 2107, GLP-1, hGIP and mouse GIP (mGIP) at each of the respective (hGLP-1), (mGLP-1), hGIP and (mGIP) receptors. As used herein OEG = mPEG.

[0036] Fig. 11 provides body weight lowering (experiment BWB-30) as a percentage for subcutaneous injection of a GLP-l / GIP co-agonist (MAR709), BWB3062 and a GLP-l / CEX agonist BWB 3063 each conjugated to DHEA at lysine 40. It also includes an FXR32993-439275agonist (cilofexor) conjugated to lysine 40 in a GLP-l / GIP co-agonist (MAR709), BWB 2100, relative to a GLP-l / CEX agonist BWB1060, and a GLP-l / GIP co-agonist (MAR709) BWB2050 into DOI mice at a daily dose of 10 nmol / kg for five days and subsequently for 5 additional days at 30nmol / kg.

[0037] Fig. 12 provides fasting blood glucose (experiment BWB-30) after ten days of treatment as shown in Figure 11 and overnight fasting, following last dose. The glucose is reported as mg / dl.

[0038] Fig. 13 provides body weight lowering (experiment BWB-31) as a percentage for subcutaneous injection of a GLP-l / CEX agonist BWB 1060 where the C-terminal amino acid in each peptide has been respectively conjugated to estrogen (E2) BWB 1050, or a PPAR agonist (GW) BWB2081, or dexamethasone (Dexa) BWB3052. Peptides were injected in DOI mice at a daily dose of 10 nmol / kg for five days and subsequently for 5 additional days at 30nmol / kg.

[0039] Fig. 14 provides fasting blood glucose (experiment BWB-31) after ten days of treatment as shown in Figure 13 and overnight fasting, following last dose. The glucose is reported as mg / dl.

[0040] Fig. 15 provides body weight lowering (experiment BWB-32) as a percentage for subcutaneous injection of a GLP-l / CEX agonist dimers BWB10080 where the C-terminal amino acid in each dimer has been respectively modified. BWB 10084 has estrogen (E2) conjugated to a single C-terminal lysine residue in the dimer. BWB10085 has a PPAR agonist (GW) conjugated to a single C-terminal lysine residue in the dimer. BWB10086 has an estrogen conjugated to a single C-terminal lysine residue in the dimer, and the other lysine is conjugated with a PPAR agonist (GW). BWB10087 has an estrogen conjugated to each C-terminal lysine residue in the dimer. BWB 10088 has a PPAR agonist (GW) conjugated to each C-terminal lysine residue in the dimer. Peptides were injected in DOI mice at a daily dose of 10 nmol / kg for five days and subsequently for 5 additional days at 30nmol / kg.

[0041] Fig. 16 provides body weight lowering (experiment BWB-34) as a percentage for subcutaneous injection of a GLP-l / CEX or GIP agonist where the C-terminal amino acid in each peptide was chemically modified. BWB2128 represents the GIP agonist with no modification to the C-terminal lysine residue. The GIP agonist conjugated at the C-terminal lysine with estrogen is numbered BWB2129. The GIP agonist conjugated at the C-terminal lysine with PPAR agonist (GW) is numbered BWB2120. The GIP agonist conjugated at the C-terminal lysine with dexamethasone is numbered BWB3070. BWB1060 represents the GLP-32993-4392751 / CEX agonist with no modification to the C-terminal lysine residue. The GLP-l / CEX agonist conjugated at the C-terminal lysine with estrogen is numbered BWB1050. The GLP-l / CEX agonist conjugated at the C-terminal lysine with PPAR agonist (GW) is numbered BWB2081. The GLP-l / CEX agonist conjugated at the C-terminal lysine with dexamethasone is numbered BWB3052. Peptides were injected in DOI mice at a daily dose of 10 nmol / kg for five days and subsequently for 5 additional days at 30nmol / kg.

[0042] Fig. 17 provides fasting blood glucose (experiment BWB-34) after ten days of treatment as shown in Figure 16 and overnight fasting, following last dose. The glucose is reported as mg / dl.

[0043] Fig. 18 provides body weight lowering (experiment BWB-44) as a percentage for subcutaneous injection of a GLP-l / CEX or GIP agonist where the C-terminal amino acid in each peptide was chemically modified. The peptides were administered as either a single peptide or as noted an equal mixture of two peptides. BWB2128 represents the GIP agonist with no modification to the C-terminal lysine residue. The GIP agonist conjugated at the C-terminal lysine with PPAR agonist (GW) is BWB2120. BWB1060 represents the GLP-l / CEX agonist with no modification to the C-terminal lysine residue. The GLP-l / CEX agonist conjugated at the C-terminal lysine with PPAR agonist (GW) is BWB2081. The GLP-l / CEX-GIP coagonist (MAR709) without any modification at the C-terminal lysine is BWB2050, with PPAR agonist (GW) conjugated at this amino acid it is BWB2069. Peptides were injected in DOI mice at a daily dose of 10 nmol / kg for five days and subsequently for 5 additional days at 30nmol / kg.

[0044] Fig. 19 provides body weight lowering (experiment BWB-51) as a percentage for subcutaneous injection of a series of PPAR-conjugated peptide agonists. The peptide agonists included modifications at the C-terminal amino acid in each peptide. Two different PPAR agonists were assessed, they include lanifibrinor (Lani) and the Glaxo-based agonist (GW). Four different targeting peptides were studied. They included selective agonists at the GLP-1, GIP and Glucagon (GCG) receptors, as well as coagonists that the GLP-l / GIP receptors and the GCG / GIP receptors, and a triple agonist at all three receptors (GLP-l / GIP / GCG). The lanifibrinor conjugates include BWB2145 (GIP / GCG coagonist), BWB2051 (GIP / GLP-1) coagonist, BWB2082 (GLP-1 agonist), BWB2146 (GIP agonist). BWB 2135 (GCG agonist), and BWB2147 (GIP / GCG / GLP-1) triple agonist. The two GW-PPARs agonists are BWB2138 (GIP / GCG) coagonist and BWB2069 (GIP / GLP-1) coagonist. Peptides were injected in DOI mice at a daily dose of 10 nmol / kg for five days and subsequently for 5 additional days at 30nmol / kg.32993-439275

[0045] Fig. 20 presents a platform for preparation of multimeric peptide-nuclear hormone conjugates. They were prepared by conjugation of peptide agonists to a dendritic polymeric core composed of polyethylene glycol. The platform provides nearly infinite possibilities to create multimeric peptides that contain defined mixtures of different peptide and nuclear hormone pharmacology, integrated with chemical protractors to control biodistribution and pharmacokinetics. The chemical conjugation can be achieved by a host of chemistries that are chemically orthogonal to the peptides. In this instance the application of click chemistry employs a peg-polymer with four azido groups crosslinks to a heterocyclic peptide polymer by virtue of a C-terminal allelic functional group at the C-terminal end of the peptide.

[0046] Fig. 21 represents body weight lowering (experiment BWB-55) as a percentage for subcutaneous injection of a series of multimeric peptide agonists, some including PPAR agonists. BWB2081 is a GLP-l / CEX agonist that includes a C-terminally conjugated PPAR agonist (GW) at the terminal lysine side chain. Peptides BWB 10111 through 10114 represents monomeric and dimeric forms of the BWB2081 where the inclusion of carbohydrate increases the biophysical properties of the peptides, notably solubility. The dimeric analogs include two PPARs with either one or two carbohydrates. Peptide 2152 is a GLP-l / CEX agonist that is chemically modified to include a PPAR agonist and a lOkd peg-polymer as an alternative means to increase physical size and aqueous solubility. The tetrameric form of this peptide conjugate is BWB2154, and it was assembled by the approach presented in Figure 20. Peptides were injected in DOI mice at a daily dose of 10 nmol / kg for ten days.

[0047] Fig. 22 represents body weight lowering (experiment BWB-57) as a percentage for subcutaneous injection of a series of PPAR and estrogen conjugated peptide agonists. These include triple agonists at the GLP-l / GIP / GCG receptors (4301), and coagonists at the GLP-1 / GIP (Mar709) and GIP-GCG receptors (4301, 17). The Lani-conjugates include at BWB2147 (GLP-l / GIP / GCG), BWB2051 (GLP-l / GIP) and BWB2145 (GIP / GCG). The estrogen-conjugates include at BWB1075 (GLP-l / GIP / GCG), BWB1066 (GLP-l / GIP) and BWB2143 (GIP / GCG). The peptide agonists without chemical conjugation are BWB2158 (GLP-l / GIP / GCG), BWB2050 (GLP-l / GIP) and BWB2130 (GIP / GCG). Peptides were injected in DOI mice at a daily dose of 10 nmol / kg for five days. 30 nmol / kg for an additional five days, and 100 nmol / kg for the last five days.

[0048] Fig. 23 represents the uterine analysis of ovariectomized (ovx) mice that had been treated each day with GLP-1 / GIP-E2 coagonists for 21 days. The control treatment was PBS in ovx-mice and sham prepared mice. The pharmacological treatments included unopposed estrogen (E2), a fully functional GLP-l / GIP coagonist that was stably conjugated to estrogen to32993-439275prevent plasma release, BWB 1066. A chemical comparable GLP-l / GIP coagonist that was stably conjugated with estrogen but through a site-specific mutation the peptide receptor co-agonism was lowered to less than 1% of the comparator (BWB 1066), it is BWB 1073. A third conjugate was BWB 1074 and it included a fully active peptide and E2 but was chemically assembled in a form that quickly dissociates in vivo to release a fully active GLP-l / GIP coagonist and a non-specific estrogen, BWB1074. The last peptide was BWB2050 and it is a GLP-l / GIP coagonist to which no chemical modification has been made. Peptides were injected into lean, ovx-mice at a daily dose that increased every three days starting at lOnmol / kg. followed by 30, 100, 300 and eventually 1,000 nmol / kg.

[0049] Fig. 24 represents the analysis of the thymus and adrenal glands from lean, male SD-rats that had been treated with a GLP-1 / GIP-E2 coagonists for 15 days. The control treatment was PBS in healthy, lean rats. The pharmacological treatments included unopposed dexamethasone (Dexa), a fully functional coagonist that was stably conjugated to dexamethasone to prevent plasma release. BWB 3059 and a comparable peptide that was without dexamethasone (BWB2050). Peptides were injected at a daily dose that increased every three days starting at 30nmol / kg, followed by 100, and eventually 300 nmol / kg.

[0050] Fig. 25 represents the analysis of the thymus and spleen from lean, male SD-rats that had been treated with a GLP-l / GIP-Lani coagonists for 15 days. The control treatment was PBS in healthy, lean rats. The pharmacological treatments included unopposed PPAR (Lani), a fully functional coagonist that was stably conjugated to lanifibrinor to prevent plasma release. BWB 2051 and a comparable peptide that was without lanifibrinor (BWB2050). Peptides were injected at a daily dose that increased every three days starting at 30nmol / kg, followed by 100, 300, and eventually 900 nmol / kg.

[0051] Figs. 26A & 26B represent the analysis of paws in male DBA / l-mice that had been treated with a GLP-l / GIP-Dexa coagonists for 14 days, following collagen-induced arthritis. The control treatment was PBS in healthy, lean rats. The pharmacological treatments included unopposed dexamethasone at a dose of 500nmol / kg, a fully functional coagonist that was stably conjugated to dexamethasone to prevent plasma release, BWB 3059 and a comparable peptide that was without dexamethasone (BWB2050). Peptides were injected at a daily dose of 30nmol / kg, and treatment started at first signs of inflamed paws, approximately a week after autoimmunization with collagen. Fig. 26A provides the data as raw % vs vehicle and Fig. 26B presents the data as AUC % vs vehicle.

[0052] Figure 27 BWB10111 is a GLP-l / CEX agonist that has been chemically modified at the C-terminal lysine through PPAR (GW) conjugation. Mid-sequence, amino acid 24 is32993-439275modified with additional of a carbohydrate to increase aqueous solubility. The lysine at residue 12 is extended with a mini-peg linker and a terminal gamma-glutamic acid that is acetylated.

[0053] Figure 28 BWB10112 is a GLP-l / CEX agonist that has been chemically modified at the C-terminal lysine through PPAR (GW) conjugation. At the C-terminus intermediate to the lysine sidechain and the PPAR (GW) is inserted a carbohydrate to increase aqueous solubility. The lysine at residue 12 is extended with a mini-peg linker and a terminal gamma-glutamic acid that is acetylated.

[0054] Figure 29 BWB10113 is a dimer of a GLP-l / CEX agonist (BWB10111). The lysine at residue 12 is conjugated to a cysteine at the same position of a comparable form of BWB10111 to form a thioether. The dimer includes two mole equivalents of PPAR(GW) and carbohydrate per mole of peptide dimer.

[0055] Figure 30 BWB10114 is a dimer of a GLP-l / CEX agonist (BWB10111). The lysine at residue 12 is conjugated to a cysteine at the same position of a comparable form of BWB10111 to form a thioether. In contrast to BWB10113 the dimer has only a single molar equivalent of PPAR (GW) but includes two mole equivalents of carbohydrate per mole of peptide dimer.DETAILED DESCRIPTION OF THE INVENTION DEFINITIONS

[0056] In describing and claiming the invention, the following terminology will be used in accordance with the definitions set forth below.

[0057] The term "about" as used herein means greater or lesser than the value or range of values stated by 10 percent, but is not intended to designate any value or range of values to only this broader definition. Each value or range of values preceded by the term "about" is also intended to encompass the embodiment of the stated absolute value or range of values.

[0058] As used herein, the term “pharmaceutically acceptable carrier” includes any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions such as an oil / water or water / oil emulsion, and various types of wetting agents. The term also encompasses any of the agents approved by a regulatory agency of the US Federal government or listed in the US Pharmacopeia for use in animals, including humans.32993-439275

[0059] As used herein the term "pharmaceutically acceptable salt" refers to salts of compounds that retain the biological activity of the parent compound, and which are not biologically or otherwise undesirable. Many of the compounds disclosed herein are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0060] Pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases, include by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines. Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid, and the like.

[0061] As used herein, the term "treating" includes alleviation of the symptoms associated with a specific disorder or condition and / or preventing or eliminating said symptoms. For example, as used herein the term "treating diabetes" will refer in general to altering glucose blood levels in the direction of normal levels and may include increasing or decreasing blood glucose levels depending on the given situation.

[0062] As used herein, an "effective" amount or a "therapeutically effective amount" of a glucagon peptide refers to a nontoxic but sufficient amount of the peptide to provide the desired effect. For example, one desired effect would be the prevention or treatment of hyperglycemia, as measured, for example, by a decrease in blood glucose level. An alternative desired effect for the glucagon peptides of the present disclosure would include treating hypoglycemia, e g., as measured by a change in blood glucose level closer to normal, or inducing weight loss / preventing weight gain, e.g., as measured by reduction in body weight, or preventing or reducing an increase in body weight, or normalizing body fat distribution. The amount that is "effective" will vary from subject to subject, depending on the age and general condition of the individual, mode of administration, and the like. Thus, it is not always possible to specify an exact "effective amount." However, an appropriate "effective" amount in any individual case may be determined by one of ordinary skill in the art using routineexperimentation.32993-439275

[0063] The term, "parenteral" means not through the alimentary canal but by some other route, e.g., subcutaneous, intramuscular, intraspinal, or intravenous.

[0064] As used herein the term "patient" without further designation is intended to encompass any warm-blooded vertebrate domesticated animal (including for example, but not limited to livestock, horses, cats, dogs and other pets), mammals, and humans.

[0065] The term “isolated” as used herein means having been removed from its natural environment. In some embodiments, the analog is made through recombinant-DNA methods, and the analog is isolated from the host cell.

[0066] The term “purified,” as used herein relates to the isolation of a molecule or compound in a form that is substantially free of contaminants normally associated with the molecule or compound in a native or natural environment and means having been increased in purity as a result of being separated from other components of the original composition. The term "purified polypeptide" is used herein to describe a polypeptide which has been separated from other compounds including, but not limited to nucleic acid molecules, lipids and carbohydrates.

[0067] As used herein, the term "peptide" encompasses a sequence of 2 or more amino acids and typically less than 50 amino acids, wherein the amino acids are naturally occurring or coded or non-naturally occurring or non-coded amino acids. Non-naturally occurring amino acids refer to amino acids that do not naturally occur in vivo but which, nevertheless, can be incorporated into the peptide structures described herein. “Non-coded” as used herein refer to an amino acid that is not an L-isomer of any of the following 20 amino acids: Ala, Cys, Asp, Glu, Phe, Gly, His, He, Lys, Leu, Met, Asn, Pro, Gin, Arg, Ser, Thr, Vai, Trp, Tyr.

[0068] As used herein, “partly non-peptidic” refers to a molecule wherein a portion of the molecule is a chemical compound or substituent that has biological activity and that does not comprise a sequence of amino acids.

[0069] As used herein, the terms "polypeptide" and "protein" are terms that are used interchangeably to refer to a polymer of amino acids, without regard to the length of the polymer. Typically, polypeptides and proteins have a polymer length that is greater than that of "peptides." In some instances, a protein comprises more than one polypeptide chain covalently or noncovalently attached to each other.

[0070] Throughout the application, all references to a particular amino acid position by number (e.g., position 28) refer to the amino acid at that position in native glucagon (SEQ32993-439275ID NO: 1) or the corresponding amino acid position in any analogs thereof. For example, a reference herein to “position 28"’ would mean the corresponding position 27 for an analog of glucagon in which the first amino acid of SEQ ID NO: 1 has been deleted. Similarly, a reference herein to “position 28” would mean the corresponding position 29 for an analog of glucagon in which one amino acid has been added before the N-terminus of SEQ ID NO: 1.

[0071] As used herein an “amino acid modification” refers to (i) a substitution or replacement of an amino acid of the reference peptide (e.g. SEQ ID NO: 1) with a different amino acid (naturally-occurring or coded or non-coded or non-naturally-occurring amino acid), (ii) an addition of an amino acid (naturally-occurring or coded or non-coded or non-naturally-occurring amino acid) to the reference peptide or (lii) a deletion of one or more amino acids from the reference peptide.

[0072] In some embodiments, the amino acid substitution or replacement is a conservative amino acid substitution, e.g., a conservative substitution of the amino acid at one or more of positions 1, 2, 5, 7, 8, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 24, 27, 28 or 29. As used herein, the term "conservative amino acid substitution" is the replacement of one amino acid with another amino acid having similar properties, e.g., size, charge, hydrophobicity, hydrophilicity, and / or aromaticity, and includes exchanges within one of the following five groups:I. Small aliphatic, nonpolar or slightly polar residues:Ala, Ser, Thr, Pro, Gly;II. Polar, negative- charged residues and their amides and esters:Asp, Asn. Glu, Gin, cysteic acid and homocysteic acid;III. Polar, positive- charged residues:His, Arg, Lys; Ornithine (Om)IV. Large, aliphatic, nonpolar residues:Met, Leu. He. Vai, Cys, Norleucine (Nle), homocysteineV. Large, aromatic residues:Phe, Tyr, Trp, acetyl phenylalanine

[0073] In some embodiments, the amino acid substitution is not a conservative amino acid substitution, e.g., is anon-conservative amino acid substitution.

[0074] As used herein the term "amino acid" encompasses any molecule containing both amino and carboxyl functional groups, wherein the amino and carboxyl groups are attached to the same carbon (the alpha carbon). The alpha carbon optionally may have one or two further organic substituents. For the purposes of the present disclosure designation of an32993-439275amino acid without specifying its stereochemistry is intended to encompass either the L or D form of the amino acid, or a racemic mixture. However, in the instance where an amino acid is designated by its three letter code and includes a superscript number (i.e., Lys’1). such a designation is intended to specify the native L form of the amino acid at position -1 relative to native glucagon (i.e., representing an amino acid added to the N-terminus of native glucagon), whereas the D form will be specified by inclusion of a lower case d before the three letter code and superscript number (i.e., dLys'1).

[0075] As used herein a "linker" is a bond, molecule or group of molecules that binds two separate entities to one another. Linkers may provide for optimal spacing of the two entities or may further supply a labile linkage that allows the two entities to be separated from each other. Labile linkages include photocleavable groups, acid-labile moieties, base-labile moieties and enzyme-cleavable groups.

[0076] As used herein a "dimer" is a complex comprising two subunits covalently bound to one another via a linker. The term dimer, when used absent any qualifying language, encompasses both homodimers and heterodimers. A homodimer comprises two identical subunits, whereas a heterodimer comprises two subunits that differ, although the two subunits are substantially similar to one another.

[0077] As used herein, a ‘“spacer" is a molecule or group of molecules that extends the distance between two linked moieties.

[0078] As used herein the general term "polyethylene glycol chain" or " PEG chain", refers to mixtures of condensation polymers of ethylene oxide and water, in a branched or straight chain, represented by the general formula H(OCH2CH2)nOH, wherein n is at least 2. Absent any further characterization, the term is intended to include polymers of ethylene glycol with an average total molecular weight selected from the range of 500 to 40,000 Daltons, "polyethylene glycol chain" or " PEG chain" is used in combination with a numeric suffix to indicate the approximate average molecular weight thereof. For example, PEG-5,000 (or PEG 5K) refers to polyethylene glycol chain having a total molecular weight average of about 5,000 daltons.

[0079] As used herein the term "pegylated" and like terms refer to a compound that has been modified from its native state by linking a polyethylene glycol chain to the compound. A "pegylated glucagon peptide" is a glucagon peptide that has a PEG chain covalently bound to the glucagon peptide.

[0080] As used herein the term “miniPEG” or ' OEG" defines a functionalized polyethylene compound comprising the structure:32993-439275

[0081] As used herein an "‘acylated” amino acid is an amino acid comprising an acyl group which is non-native to a naturally occurring amino acid, regardless by the means by which it is produced. Exemplary methods of producing acylated amino acids and acylated peptides are known in the art and include acylating an amino acid before inclusion in the peptide or peptide synthesis followed by chemical acylation of the peptide. In some embodiments, the acyl group causes the peptide to have one or more of (i) a prolonged half-life in circulation, (ii) a delayed onset of action, (iii) an extended duration of action, (iv) an improved resistance to proteases, such as DPP-IV, and (v) increased potency at the glucagon superfamily peptide receptor.

[0082] As used herein, an “alkylated” amino acid is an amino acid comprising an alkyl group which is non-native to a naturally occurring amino acid, regardless of the means by which it is produced. Exemplary methods of producing alkylated amino acids and alkylated peptides are known in the art and include alkylating an amino acid before inclusion in the peptide or peptide synthesis followed by chemical alkylation of the peptide. Without being held to any particular theory, it is believed that alkylation of peptides will achieve similar, if not the same, effects as acylation of the peptides, e.g., a prolonged half-life in circulation, a delayed onset of action, an extended duration of action, an improved resistance to proteases, such as DPP-IV. and increased potency at the glucagon superfamily peptide receptor.

[0083] The term " Ci-Cnalkyl" wherein n can be from 1 through 18, as used herein, represents a branched or linear alkyl group having from one to the specified number of carbon atoms. For example. Ci-Ce alkyl represents a branched or linear alkyl group having from 1 to 6 carbon atoms. Typical Ci-Cis alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl and the like. Alkyl groups optionally can be substituted, for example, with hydroxy (OH), halo, aryl, carboxyl, thio, Cs-Cs cycloalkyl, and amino.

[0084] The term " Co-Cnalkyl" wherein n can be from 1-18, as used herein, represents a branched or linear alkyl group having up to 18 carbon atoms. For example, the term “(Co-Cs alkyl)OH” represents a hydroxyl parent moiety attached to an alkyl substituent having up to 6 carbon atoms (e g. -OH. -CH2OH, -C2H4OH. -CsHsOH, -C4H8OH. -C5H10OH. -C6H12OH).32993-439275

[0085] The term " C2-Cnalkenyl" wherein n can be from 2 through 18, as used herein, represents an unsaturated branched or linear group having from 2 to the specified number of carbon atoms and at least one double bond. Examples of such groups include, but are not limited to, 1-propenyl, 2-propenyl (-CH2-CH=CH2), 1,3-butadienyl, (-CH=CHCH=CH2), 1-butenyl (-CH=CHCH2CH3), hexenyl, pentenyl, and the like. Alkenyl groups optionally can be substituted, for example, with hydroxy (OH), halo, aryl, carboxyl, thio, Cs-Cs cycloalkyl, and amino.

[0086] The term " C2-C11 alkynyl" wherein n can be from 2 to 18, refers to an unsaturated branched or linear group having from 2 to n carbon atoms and at least one triple bond. Examples of such groups include, but are not limited to, 1-propynyL 2-propynyl, 1-butynyl, 2-butynyl, 1 -pentynyl, and the like. Alkynyl groups optionally can be substituted, for example, with hydroxy (OH), halo, aryl, carboxyl, thio, C3-C8 cycloalkyl, and amino.

[0087] As used herein the term "aryl" refers to a monocyclic or polycyclic (for example, bicyclic, tricyclic, or tetracyclic) aromatic groups. The size of the ary 1 ring or rings is indicated by designating the number of carbons present. For example, the term "(C1-C3 alkyl)(Ce-Cio aryl)" refers to a 6 to 10 membered aryl that is attached to a parent moiety via a one to three membered alkyl chain. Unless otherwise indicated, an aryl group can be unsubstituted or substituted with one or more, and in particular one to five groups independently selected from, for example, halo, alkyl, alkenyl, OCF3, NO2, CN, NC. OH, alkoxy, amino. CO2H. C3-C8 cycloalkyl, C(O)Oalkyl. aryl, and heteroaryl. Exemplary aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, chlorophenyl, indanyl, indenyl, methylphenyl, methoxyphenyl, trifluoromethylphenyl, nitrophenyl, 2,4-methoxy chlorophenyl, and the like.

[0088] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic ring system containing one or more aromatic rings and containing at least one nitrogen, oxygen, or sulfur atom in an aromatic ring. The size of the heteroaryl ring and the presence of substituents or linking groups are indicated by designating the number of carbons present. For example, the term "(Ci-Ce alkyl)(Cs-C6 heteroaryl)" refers to a 5 or 6 membered heteroaryl that is attached to a parent moiety via a one to 6 membered alkyl chain. Unless otherwise indicated, a heteroaryl group can be unsubstituted or substituted with one or more, and in particular one to five groups independently selected from, for example, halo, alkyl, alkenyl, OCF3, NO2, CN, NC, OH, alkoxy, amino, CO2H, C3-C8 cycloalkyl, C(O)Oalkyl, aryl, and heteroaryl. Examples of heteroaryl groups include, but are not limited to, thienyl, furyl, pyridyl, oxazolyl, quinolyl,32993-439275thiophenyl, isoquinolyl, indolyl, triazinyl, triazolyl, isothiazolyl, isoxazolyl, imidazolyl, benzothiazolyl, pyrazinyl, pyrimidinyl, thiazolyl, and thiadiazolyl.

[0089] As used herein, the term “heteroalkyl” refers to a linear or branched hydrocarbon containing the indicated number of carbon atoms and at least one heteroatom in the backbone of the structure. Suitable heteroatoms for purposes herein include but are not limited to N, S, and O. Heteroalkyl groups optionally can be substituted, for example, with hydroxy (OH), halo, aryl, carboxyl, and amino.

[0090] As used herein, the term “halogen’’ or “halo” refers to one or more members of the group consisting of fluorine, chlorine, bromine, and iodine.

[0091] As used herein, the term “incretin peptide” refers to those peptides which have biological activity (as agonists or antagonists) at any one or more of the glucagon, GLP-1, GLP-2, and GIP receptors and comprise an amino acid sequence that shares at least 40% sequence identity (e.g., 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%) with at least one of native glucagon, native oxyntomodulin, native exendin-4, native GLP-1, native GLP-2, or native GIP. Unless otherwise stated, any reference to an amino acid position in an incretin peptide (e.g. for linkage of aNHR ligand, a conjugate moiety, a hydrophilic polymer, acylation or alkylation or forming multimers) refers to the corresponding position relative to the native glucagon amino acid sequence (SEQ ID NO: 1).

[0092] As used herein, the term “selectivity” of a molecule for a first receptor relative to a second receptor refers to the following ratio: EC so of the molecule at the second receptor divided by the ECso of the molecule at the first receptor. For example, a molecule that has an ECso of 1 nM at a first receptor and an ECso of 100 nM at a second receptor has 100-fold selectivity for the first receptor relative to the second receptor.

[0093] The term "identity" as used herein relates to the similarity between two or more sequences. Identity is measured by dividing the number of identical residues by the total number of residues and multiplying the product by 100 to achieve a percentage. Thus, two copies of exactly the same sequence have 100% identity, whereas two sequences that have amino acid deletions, additions, or substitutions relative to one another have a lower degree of identity. Those skilled in the art will recognize that several computer programs, such as those that employ algorithms such as BLAST (Basic Local Alignment Search Tool, Altschul et al. (1993) J. Mol. Biol. 215:403-410) are available for determining sequence identity.

[0094] As used herein, the term “glucagon superfamily peptide” refers to a group of peptides related in structure in their N-terminal and C-terminal regions (see, for example,32993-439275Sherwood et al., Endocrine Reviews 21: 619-670 (2000)). Members of this group include all incretin peptides, as well as Growth Hormone Releasing Hormone (GHRH; SEQ ID NO: 7), vasoactive intestinal peptide (VIP; SEQ ID NO: 8), pituitary adenylate cyclase-activating polypeptide 27 (PACAP-27; SEQ ID NO: 9), peptide histidine isoleucine (PHI; SEQ ID NO: 12), peptide histidine methionine (PHM; SEQ ID NO: 10), secretin (SEQ ID NO: 11), and analogs, derivatives or conjugates with up to 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid modifications relative to the native peptide. Such peptides preferably retain the ability to interact (agonist or antagonist) with receptors of the glucagon receptor superfamily. Unless otherwise stated, any reference to an amino acid position in a glucagon superfamily peptide (e.g. for linkage of aNHR ligand, a conjugate moiety, a hydrophilic polymer, acylation or alkylation) refers to the corresponding position relative to the native glucagon amino acid sequence (SEQ ID NO: 1), see Figure 1 for an alignment of representative glucagon superfamily peptides.

[0095] The term “glucagon agonist peptide7' refers to a compound that binds to and activates downstream signaling of the glucagon receptor. However, this term should not be construed as limiting the compound to having activity at only the glucagon receptor. Rather, the glucagon agonist peptides of the present disclosures may exhibit additional activities at other receptors, as further discussed herein. Glucagon agonist peptides, for example, may exhibit activity (e.g., agonist activity) at the GLP-1 receptor and / or the GIP receptor. Also, the term “glucagon agonist peptide’7should not be construed as limiting the compound to only peptides. Rather, compounds other than peptides are encompassed by this term. Accordingly, the glucagon agonist peptide in some respects is a peptide in conjugate form (a heterodimer, a multimer, a fusion peptide), a chemically-derivatized peptide, a pharmaceutical salt of a peptide, a peptidomimetic, and the like.

[0096] The term “GLP-1 agonist peptide" refers to a compound that binds to and activates downstream signaling of the GLP-1 receptor. However, this term should not be construed as limiting the compound to having activity at only the GLP-1 receptor. Rather, the GLP-1 agonist peptides of the present disclosures may exhibit additional activities at other receptors, as further discussed herein. GLP-1 agonist peptides, for example, may exhibit activity (e.g., agonist activity) at the glucagon receptor and / or the GIP receptor. Also, the term “GLP-1 agonist peptide” should not be construed as limiting the compound to only peptides. Rather, compounds other than peptides are encompassed by this term. Accordingly, the GLP-1 agonist peptide in some respects is a peptide in conjugate form (a heterodimer, a multimer, a32993-439275fusion peptide), a chemically-derivatized peptide, a pharmaceutical salt of a peptide, a peptidomimetic, and the like.

[0097] The term “GIP agonist peptide” refers to a compound that binds to and activates downstream signaling of the GIP receptor. However, this term should not be construed as limiting the compound to having activity at only the GIP receptor. Rather, the GIP agonist peptides of the present disclosures may exhibit additional activities at other receptors, as further discussed herein. GIP agonist peptides, for example, may exhibit activity (e.g., agonist activity) at the GLP-1 receptor. Also, the term ‘‘GIP agonist peptide” should not be construed as limiting the compound to only peptides. Rather, compounds other than peptides are encompassed by this term. Accordingly, the GIP agonist peptide in some respects is a peptide in conjugate form (a heterodimer, a multimer, a fusion peptide), a chemically-derivatized peptide, a pharmaceutical salt of a peptide, a peptidomimetic, and the like.

[0098] As used herein the term “native glucagon” refers to a peptide consisting of the sequence of SEQ ID NO: 1.

[0099] As used herein, the term “native GLP-1” is a generic term that designates GLP-l(7-36) amide (SEQ ID NO: 38), GLP-l(7-37) acid (SEQ ID NO: 2) or a mixture of those two compounds.

[0100] As used herein, the term “native GIP” refers to a peptide consisting of SEQ ID NO: 4.

[0101] As used herein, "nuclear hormone receptor ligand" or “NHR ligand” refers to a hydrophobic or lipophilic moiety that has biological activity (either agonist or antagonist) at a nuclear hormone receptor (NHR). The NHR ligand is wholly or partly non-peptidic. In some embodiments, the NHR ligand is an agonist that binds to and activates the NHR.Examples of NHR ligands include PPAR agonists, Famesoid X receptor agonists and steroids.

[0102] As used herein the term, “nuclear hormone receptors” (NHRs) identifies any ligand-activated protein that regulates gene expression within the cell nucleus, sometimes in concert with other co-activators and co-repressors. Type I nuclear receptors are typically found in the cytoplasm and translocate to the nucleus upon ligand binding, while Type II receptors reside in the nucleus.

[0103] As used herein, "peroxisome proliferator activated receptors" (PPARs) are a family of nuclear hormone receptors that regulate genes involved in lipid and glucose metabolism, energy homeostasis, and inflammation. They consist of three subtypes: PPARa, PPAR0 / 5, and PPARy. A " PPAR agonist" is any compound that activates one or more of the32993-439275PPAR receptors selected from PPARa, PPARp / 5, and PPARy. A " PPAR co-agonist" is any compound that activates both PPARa, and PPAR0 / 5. A "pan-PPAR agonist" is any compound that activates all PPAR receptors to a variable degree.

[0104] As used herein a generic reference to a “steroid’' defines a set of compounds, either naturally occurring or synthesized, having a structure of Formula A:Formula Awherein R1and R2, when present, are independently moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula A to a nuclear hormone receptor; R3and R4are independently moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula A to a nuclear hormone receptor; and each dashed line represents an optional double bond. Formula A may further comprise one or more substituents at one or more of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 14, 15, 16, and 17. Contemplated optional substituents include, but are not limited to, OH, NH2, ketone, and Ci-Cis alkyl groups. Specific, nonlimiting examples of steroids and derivatives thereof include cholesterol, cholic acid estradiol, testosterone, and hydrocortisone.

[0105] As used herein, “bile acids and derivatives thereof’ refers to compounds, either naturally occurring or synthesized, of Formula G:Formula Gwherein each of R15, R16, and R17are independent moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula M to a nuclear hormone receptor. In some embodiments, each of R13and R16are independently hydrogen, (Co-Cs alkyl)halo, Ci-Cis alkyl, C2-C18 alkenyl, C2-C18 alkynyl, heteroalkyl, or (Co-Cs alkyl)OH; and R17is OH, (Co-Cs alkyl)NH(Ci-C4 alkyl)SO3H, or (Co-Cs alkyl)NH(Ci-C4alkyl)COOH.Formula M may further comprise one or more substituents at one or more of positions 1, 2, 3, 4,32993-4392755, 6, 7, 8, 9, 11, 12, 14, 15, 16, and 17. Nonlimiting examples of bile acids include cholic acid, deoxy cholic acid, lithocholic acid, chenodeoxy cholic acid, taurocolic acid, and glycocholic acid.

[0106] As used herein, “cholesterol and derivatives thereof’ refers to compounds, either naturally occurring or synthesized, comprising a structure similar to that of cholesterol, as shown below:CholesterolDerivatives of cholesterol can include oxysterols, such as hydroxy cholesterol, 24(S)-hydroxy cholesterol, 27-hydroxy cholesterol, and cholestenoic acid.

[0107] As used herein, “estradiol and derivatives thereof refers to compounds, either naturally occurring or synthesized, of Formula B:Formula Bwherein R1, R5and R6are moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula B to the estrogen receptor. In some embodiments, the structure of Formula B is substituted with one or more substituents at one or more positions of the tetracyclic ring, such as, for example, positions 1, 2, 4, 6, 7, 8, 9, 11, 12, 14, 15, 16, and 17. In some cases, the substituent comprises a ketone at position-6. The structure of estradiol is provided below:32993-439275Specific, nonlimiting examples of derivatives of estradiol include include |3-estradiol 17-acetate, P-estradiol 17-cypionate, P-estradiol 17-enanthate, P-estradiol 17-valerate, P-estradiol 3,17-diacetate, P-estradiol 3.17-dipropionate, P-estradiol 3-benzoate, P-estradiol 3-benzoate 17-n-butyrate, P-estradiol 3-glycidyl ether, P-estradiol 3-methyl ether, P-estradiol 6-one, P-estradiol 3-glycidyl, P-estradiol 6-one 6-(O-carboxymethyloxime), 16-epiestriol, 17-epiestriol, 2-methoxy estradiol, 4-methoxy estradiol, estradiol 17-phenylpropi onate, and 17P-estradiol 2-methyl ether, 17a-ethynylestradiol, megestrol acetate, and estriol.

[0108] As used herein, “testosterone and derivatives thereof’ refers to compounds, either naturally occurring or synthesized, of Formula F:wherein R1, when present, R2, R3and R6are each independently a moiety that permits or promotes agonist or antagonist activity upon binding of the compound of Formula F to a nuclear hormone receptor; and each dashed line represents an optional double bond, with the proviso that no more than one of the optional carbon-carbon double bond is present at position 5. In some embodiments, the structure of Formula F is substituted with one or more substituents at one or more positions of the tetracyclic ring, such as, for example, positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 14, 15, 16, and 17. Specific, nonlimiting examples of derivatives of testosterone include dehydroepiandrosterone, androstenedione, 5-androstenediol, androsterone, and dihydrotestosterone.

[0109] As used herein, “fatty acids and derivatives thereof’ refers to carboxylic acids comprising a long unbranched Ci to C28 alkyl or C2 to C28 alkenyl moiety and can optionally comprise one or more halo substituents and / or optionally comprise one or more substituents other than halo. As used herein a "fatty diacid" refers to a long unbranched Ci to C28 alkyl or C2 to C28 alkenyl chain comprising two carboxylic acid groups, optionally at each end of the long alkyl or alkenyl chain. In some embodiments, the long unbranched alkyl or alkenyl moiety can be wholly halo substituted (e.g.. all hydrogens replaced with halo atoms). Specific, nonlimiting examples of fatty acids include formic acid, acetic acid, n-caproic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadeconoic acid, palmitic acid, heptadecanoic acid, stearic32993-439275acid, nonadecanoic acid, arachidic acid, heneicosanoic acid, behenic acid, tricosanoic acid, mead acid, myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, linoleic acid, a-linolenic acid, elaidic acid, petroselinic acid, arachidonic acid, dihydroxyeicosatetraenoic acid (DiHETE), octadecynoic acid, eicosatriynoic acid, eicosadienoic acid, eicosatrienoic acid, eicosapentaenoic acid, erucic acid, dihomolinolenic acid, docosatrienoic acid, docosapentaenoic acid, docosahexaenoic acid, and adrenic acid.

[0110] As used herein the term “phosphonic fatty acid” defines a lipid comprising both a carboxy acid and a phosphoric acid group wherein the carboxy acid and a phosphoric acid group are separated by a long carbon chain.

[0111] As used herein, "cortisol and derivatives thereof’ refers to compounds, either naturally occurring or synthesized, of Formula C:Formula Cwherein R2, R3, R6, R7, R8, R9, and R10are each independent moieties that permit or promote agonist or antagonist activity upon the binding of the compound of Formula C to a nuclear hormone receptor; and each dash represents an optional double bond. In some embodiments, the structure of Formula C is substituted with one or more substituents at one or more positions of the tetracyclic ring, such as, for example, positions 1, 2, 4, 5, 6, 7, 8, 11, 12, 14, and 15. Specific, nonlimiting examples of derivatives of cortisol and derivatives thereof include cortisol, cortisone acetate, beclometasone, prednisone, prednisolone, methylprednisolone, betamethasone, trimcinolone, and dexamethasone.

[0112] As used herein, the term ‘‘prodrug” is defined as any compound that undergoes chemical modification before exhibiting its full pharmacological effects.

[0113] As used herein the term "chemical cleavage" absent any further designation encompasses a non-enzymatic reaction that results in the breakage of a covalent chemical bond.EMBODIMENTS

[0114] The present disclosure provides incretin peptides conjugated with nuclear hormone receptor (NHR) ligands. In some aspects, the NHR ligands of the conjugates32993-439275disclosed herein interact with nuclear hormone receptors involved in metabolism or glucose homeostasis, wherein the conjugate provides superior biological effects on metabolism or glucose homeostasis compared to the incretin peptide alone or the NHR ligand alone. Without being bound by a theory of the invention, the NHR ligand may serve to target the incretin peptide to particular types of cells or tissues; or alternatively the incretin peptide may serve to target the NHR ligand or enhance its transport into the cell, e.g. through binding of peptide to a receptor that internalizes the conjugate.

[0115] In accordance with one embodiment, the incretin peptide / NHR ligand conjugates of the present disclosure can be represented by the following formula:Q-L-Ywherein Q is an incretin peptide, Y is a NHR ligand, and L is a linking group or a bond covalently linking Q to L.The incretin peptide (O) of the conjugate Q-L-Y

[0116] The incretin peptide (Q) in some embodiments can be a peptide that exhibits agonist activity at the glucagon (Gcg) receptor, agonist activity at the GLP-1 receptor, agonist activity at the GIP receptor, co-agonist activity at the glucagon and GLP-1 receptors, co-agonist activity at the GLP-1 and GIP receptors, or tri-agonist activity at the glucagon, GIP, and GLP-1 receptors. In some embodiments, the incretin peptide exhibits antagonist activity at the glucagon, GLP-1 and / or GIP receptor.

[0117] The incretin peptide (Q) can be any of the know n incretins, including for example, native glucagon, GLP-1. or GIP, or an agonist analog of glucagon, GLP-1, or GIP, including a glucagon / GLP-1 co-agonist. a GIP / GLP-1 co-agonist, and a glucagon / GIP / GLP-1 tri-agonist. In one embodiment the incretin peptide (Q) of the general formula: Q-L-Y is a peptide selected from Semaglutide (SEQ ID NO: 13), tirzepatide (SEQ ID NO: 14), retatrutide (SEQ ID NO: 15) and analogs thereof, wherein the NHR receptor (Y) is linked to Q via the side chain of an amino acid selected from positions 12, 17, 20, 24, 28. 30. 40. the C-terminal amino acid of the incretin peptide and any amino acid of a C-terminal extension of Q.

[0118] In one embodiment the incretin component of the Q-L-Y conjugate comprises the structure of HX2QGTFTSDYSKYLDX16RRAQX21FVKWLX27NK (SEQ ID NO: 39), wherein

[0119] X2 is Ser or dSer;

[0120] Xie is Aib;32993-439275

[0121] X21 is Glu or Asp; and

[0122] X27 is Nle or Met, wherein Y is linked to Q via the side chain of an amino acid selected from positions 24 and any amino acid of a C-terminal extension of the incretin peptide, optionally via a spacer, optionally wherein the side chain of the lysine at position 29 is acylated with a C16-C30 fatty acid, a C16-C30 phosphonic fatty acid, or a C16-C30 diacid, optionally wherein Q is GW501516.

[0123] In one embodiment the incretin component of the conjugate Q-L-Y comprises the structure ofH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIAWLVX28GRX3i (SEQ ID NO: 28), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu:X28 is Arg or Lys; andX31 is Lys, Gly, or a peptide extension comprising the sequence PSSGAPPPSK (SEQ ID NO: 29), whereinY is linked to Q via the side chain of an amino acid selected from positions 12, 20, 28, 31 or 40.

[0124] In one embodiment the incretin component of the conjugate Q-L-Y comprises the structure of YX2X3GTFTSDYSIX13LX15KIAQX20AFVQWLIAGGPSSGAPPPS-R20 (SEQ ID NO: 40), whereinX2 and X13 are each Aib;X3 is Glu or Gin;X15 is Glu or Asp;X20 is Aib; andR20 is CONH2, wherein Y is linked to Q via the side chain of an amino acid selected from positions 16, 20, the C-terminal amino acid of the incretin peptide or an amino acid added to the C-terminus of Q.

[0125] In one embodiment, the incretin component of the conjugate Q-L-Y comprises the structure of YX2X3GTX6X7SDYSIX13LX15KIAQX20AFVQWLIAGGPSSGAPPPSX40-R20 (SEQ ID NO: 16) or32993-439275 YX2X3GTX6X7SDYSIX13LX15KX17AQX200AFVQWLLEGGPSSGAPPPSX40-R20 (SEQ ID NO: 17)whereinX2 is Aib;X3 is Glu or Gin;Xe is alpha methylated Phe or Phe;X7 is isoacyl-Thr, Thr, or isoacyl-Thr acylated at the alpha amine with a selfcleaving dipeptide disclosed herein dipeptide element (A-B), wherein A is an amino acid and B is an N-alkylated amino acid;X13 is Aib or alpha methylated Leu;X15 is Glu or Asp;X20 is Lys or Aib;X200 is Aib;X40 is Lys or absent;R20 is CONH2; andX17 and X20 are independently an amino acid comprising a (C1-C4 alkyl)NH2 side chain, wherein Y is linked to Q via the side chain of an amino acid selected from positions 17, 20, 24, or 40, the C-terminal amino acid of the incretin peptide or an amino acid added to the C-terminus of Q. In one embodiment the incretin peptide comprises the sequence of SEQ ID NO: 17 wherein X2 is Aib, X3 is Glu, Xe is alpha methylated Phe, X7 is said acylated isoacyl-Thr or Thr; X13 is Aib. X15 is Glu. optionally wherein X20 is Lys acylated via its side chain with (COCH2(OCH2CH2)2NH)2-yE-COCi8H36CO2H, or Lys acylated via its side chain with (COCH2(OCH2CH2)2NH)2-YE-COCI8H36PO3H2 and R20 is CONH2. In one embodiment the incretin peptide comprises the sequence of SEQ ID NO: 17 wherein X2 is Aib, X3 is Gin, X13 is alpha methylated Leu, and X15 is Asp: X17 is Lys. optionally acylated via its side chain with (COCH2(OCH2CH2)2NH)2-yE-COCi8H36CO2H or acylated via its side chain with (COCH2(OCH2CH2)2NH)2-YE-COCI8H36PO3H2, X40 is Lys and R20 is CONH2.

[0126] In one embodiment the incretin component of the conjugate Q-L-Y comprises the structure of X1X2X3GTFTSDYSIX13LX15X16KX18QX20AFVQWLLEGGPSSGAPPPS-R20 (SEQ ID NO: 41)whereinXi is Tyr or His;X2 is Aib;32993-439275Xs is Gin or His;X13 is Tyr, (aMe)Leu, (aMe)Ile, (aMe)Met, (aMe)nLeu or Aib;X15 is Glu or Asp;Xi6is Aib, Orn or Lys;Xis is Ala or Tyr;X20 is Aib; andR20 is COOH or CONH2, wherein Y is linked to Q via the side chain of an amino acid selected from positions 17, 20, the C-terminal amino acid of the incretin peptide or an amino acid of a C-terminal extension of Q, optionally at position 40, optionally via a C-terminal extension of a Lys.

[0127] In one embodiment the incretin component of the conjugate Q-L-Y comprises the structure ofi) Y-Aib-EGTFTSDYSI-Aib-LDKI AQKAFVQWLI AGGPSSGAPPPSX10 (SEQ ID NO: 19); orii) Y-Aib-QGTFTSDYSIXisLDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSX40, (SEQ ID NO: 20), whereinXis is Leu or alpha-methyl leucine; andX40 is Lys or is absent, wherein Y is linked to Q via the side chain of an amino acid selected from positions 16, 17, 20 or 40, or an amino acid added to the C -terminus of Q.NHR agonist (Y) of the conjugate Q-L-Y

[0128] In one embodiment the NHR ligand component (Y) of the conjugate Q-L-Y is a peroxisome proliferator activated receptor (PPAR) agonist. In one embodiment, the PPAR agonist is an a PPAR agonist, including for example, pemafibrate, fenofibrate or gemfibrozil. In one embodiment, the PPAR agonist is a y PPAR agonist, including for example pioglitazone. In one embodiment, the PPAR agonist is a 5 PPAR agonist, including for example, seladelpar or GW501516. In one embodiment, the PPAR agonist is an a and y PPAR dual agonist, including for example, aleglitazar, muraglitazar, oxeglitazar, naveglitazar tesaglitazar, saroglitazar and chiglitazar. In one embodiment the PPAR agonist is selected from the group consisting of GW501516, pemafibrate, chiglitazar and saroglitzazar. In one embodiment, Y is GW501516. In one embodiment Q is not lanifibranor.

[0129] In one embodiment the NHR ligand component (Y) of the conjugate Q-L-Y is a steroid, comprising a structure as shown in Formula A:32993-439275Formula Awherein R1and R2. when present, are independently moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula A to a Type I nuclear hormone receptor; R3and R4are independently moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula A to a Type I nuclear hormone receptor; and each dashed line represents an optional double bond. Formula A may further comprise one or more substituents at one or more of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 14, 15, 16, 17, 18, and 19. Contemplated optional substituents include, but are not limited to, OH, NH2, ketone, and Ci-Cis alkyl groups.

[0130] In some embodiments, Y comprises a structure of Formula A wherein R1is present and is hydrogen, C1-C7 alkyl; (C0-C3 alkyl)C(O)Ci-C? alkyl, (C0-C3 alkyl)C(O)aiyl, or SO3H;R2is present and is hydrogen, halo, OH, or C1-C7 alkyl;R3is hydrogen, halo, OH, or C1-C7 alkyl;R4is hydrogen, (Co-Cs alkyl)halo, Ci-Cs alkyl, C2-C8 alkenyl, C2-18 alkynyl, heteroalkyl, (Co-C8alkyl)aryl, (Co-C8alkyl)heteroaryl, (Co-C8alkyl)OCi-C8alkyl, (Co-Cs alkyl)OC2-C8alkenyl, (Co-C8alkyl)OC2-Cs alkynyl, (Co-C8alkyl)OH, (Co-C8alkyl)SH, (Co-C8alkyl)NR24Ci-C8alkyl, (Co-C8alkyl)NR24C2-C8alkenyl, (Co-C8alkyl)NR24C2-Cs alkynyl, (Co-C8alkyl)NR24H2, (Co-Cs alkyl)C(O)Ci-Cs alkyl, (Co-C8alkyl)C(O)C2-C8alkenyl, (Co-C8alkyl)C(O)C2-Cs alkynyl, (Co-C8alkyl)C(O)H, (Co-C8alkyl)C(O)aryl, (Co-Cs alkyl)C(O)heteroaryl, (Co-Cs alkyl)C(O)OCi-Cs alkyl, (Co-C8alkyl)C(O)OC2-C8alkenyl, (Co-Cs alkyl)C(O)OC2-Cs alkynyl, (Co-Cs alkyl)C(O)OH. (Co-Cs alkyl)C(O)O aryl, (Co-Cs alkyl)C(O)O heteroaryl, (Co-Cs alkyl)OC(O)Ci-C8 alkyl, (Co-Cs alkyl)OC(O)C2-Cs alkenyl, (Co-Cs alkyl)OC(O)C2-Ci8 alkynyl, (Co-Cs alkyl)C(O)NR24Ci-C8alkyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkenyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkynyl, (Co-Cs alkyl)C(O)NR24H2, (Co-Cs alkyl)C(O)NR24aryl. (Co-Cs alkyl)C(O)NR24heteroaryl, (Co-Cs alkyl)NR24C(O)Ci-Cs alkyl, (Co-Cs alkyl)NR24C(O)C2-Cs alkenyl, or (Co-Cs alkyl)NR24C(O)C2-Cs alkynyl, (Co-Cs alkyl)NR24C(O)OH, (Co-Cs alkyl)OC(O)OCi-Cs alkyl, (Co-Cs alkyl)OC(O)OC2-Cs alkenyl, (Co-Cs alkyl)OC(O)OC2-Cs alkynyl, (Co-Cs alkyl)OC(O)OH, (Co-Cs alkyl)OC(O)NR24Ci-Cs alkyl,32993-439275(Co-C8alkyl)OC(O)NR24C2-C8alkenyl, (Co-Cs alkyl)OC(O)NR24C2-C8 alkynyl, (Co-C8alkyl)OC(O)NR24H2, (Co-C8alkyl)NR24(O)OCi-C8alkyl, (Co-Cs alkyl)NR24(O)OC2-C8alkenyl, (Co-C8aJkyl)NR24(O)OC2-Cs alkynyl, or (Co-C8alkyl)NR24(O)OH; and,R24is hydrogen or C1-C7 alkyl.

[0131] In some embodiments, R1is hydrogen, propionate, acetate, benzoate, or sulfate; R2is hydrogen or methyl; R3is hydrogen or methyl; and R4is acetate, cypionate, hemisucciniate, enanthate, or propionate.

[0132] In embodiments wherein Y comprises a structure of Formula A. Y is conjugated to L (e.g. when L is a linking group) or Q (e.g. when L is a bond) at any position of Formula A that is capable of reacting with Q or L. One skilled in the art could readily determine the position of conjugation on Formula A and means of conjugation of Formula A to Q or L in view of general knowledge and the disclosure provided herein. In some embodiments, Formula A is conjugated to L or Q at any of positions 1. 2, 3, 4. 5, 6, 7. 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 of Formula A. In some embodiments, Formula A is conjugated to L or Q at position 1, 3, 6, 7, 12, 10, 13, 16, 17, or 19 of Formula A.

[0133] In some embodiments, Y acts at an estrogen receptor (e.g. ERa, ERP). In some embodiments, Y permits or promotes agonist activity at the estrogen receptor, while in other embodiments Y is an antagonist of ER. In exemplary embodiments, Y can have a structure of Formula B:Formula Bwherein R1, R5and R6are moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula B to the estrogen receptor. In some embodiments, Formula B further comprises one or more substituents at one or more of positions 1, 2. 4, 6, 7, 8, 9, 11, 12, 14, 15, and 16 (e.g. a ketone at position-6).

[0134] In some embodiments, Y comprises a structure of Formula B, wherein R1is hydrogen, C1-C7 alkyl; (C0-C3 alkyl)C(O)Ci-C7 alkyl, (C0-C3 alkyl)C(O)aryl, or SO3H;R5is hydrogen, (Co-C8alkyljhalo, Ci-Cs alkyl, C2-C8alkenyl, C2-18 alkynyl. heteroalkyl, (Co-C8alky ljaryl, (Co-C8alkyl)heteroaryl, (Co-C8alkyl)OCi-C8alkyl, (Co-C8alkyl)OC2-C832993-439275alkenyl, (Co-Cs alkyl)OC2-C8alkynyl, (Co-Cs alkyl)OH, (Co-C8alkyl)SH, (Co-C8alkyl)NR24Ci-C8alkyl, (Co-Cs alkyl)NR24C2-Cs alkenyl, (Co-C8alkyl)NR24C2-Cs alkynyl, (Co-Cs alkyl)NR24H2, (Co-C8alkyl)C(O)Ci-C8alkyl, (Co-C8alkyl)C(O)C2-C8alkenyl, (Co-C8alkyl)C(O)C2-Cs alkynyl, (Co-C8alkyl)C(O)H, (Co-C8alkyl)C(O)aryl, (Co-C8alkyl)C(O)heteroaryl, (Co-C8alkyl)C(O)OCi-Cs alkyl, (Co-C8alkyl)C(O)OC2-C8alkenyl, (Co-Cs alkyl)C(O)OC2-Cs alkynyl, (Co-C8alkyl)C(O)OH, (Co-C8alkyl)C(O)O aryl, (Co-C8alkyl)C(O)O heteroaryl, (Co-C8alkyl)OC(O)Ci-Cs alkyl, (Co-C8alkyl)OC(O)C2-C8alkenyl, (Co-C8alkyl)OC(O)C2-Ci8alkynyl, (Co-C8alkyl)C(O)NR24Ci-C8alkyl. (Co-C8alkyl)C(O)NR24C2-C8alkenyl, (Co-C8alkyl)C(O)NR24C2-Cs alkynyl, (Co-Cs alkyl)C(O)NR24H2, (Co-Cs alkyl)C(O)NR24aryl, (Co-Cs alkyl)C(O)NR24heteroaryl, (Co-C8alkyl)NR24C(O)Ci-Cs alkyl, (Co-Cs alkyl)NR24C(O)C2-Cs alkenyl, or (Co-Cs alkyl)NR24C(O)C2-C8alkynyl, (Co-C8alkyl)NR24C(O)OH, (Co-Cs alkyl)OC(O)OCi-C8alkyl, (Co-Cs alkyl)OC(O)OC2-Cs alkenyl, (Co-Cs alkyl)OC(O)OC2-Cs alkynyl, (Co-Cs alkyl)OC(O)OH, (Co-C8alkyl)OC(O)NR24Ci-Cs alky 1, (Co-Cs alkyl)OC(O)NR24C2-Cs alkenyl, (Co-Cs alkyl)OC(O)NR24C2-Cs alkynyl, (Co-Cs alkyl)OC(O)NR24H2, (Co-Cs alkyl)NR24(O)OCi-Cs alkyl, (Co-Cs alkyl)NR24(O)OC2-Cs alkenyl, (Co-Cs alkyl)NR24(O)OC2-C8alkynyl, or (Co-Cs alkyl)NR24(O)OH;R6is hydrogen, Ci-Cs alkyl, C2-Cs alkenyl, C2-Cs alkynyl, heteroalkyl, (Co-Cs alkyl)aryl, (Co-Cs alkyl)heteroaryl, (Co-C8alkyl)C(O)Ci-Cs alkyl, (Co-Cs alkyl)C(O)C2-Cs alkenyl, (Co-Cs alkyl)C(O)C2-Cs alkynyl, (Co-Cs alkyl)C(O)H, (Co-Cs alkyl)C(O)aryl, (Co-Cs alkyl)C(O)heteroaryl, (Co-C8alkyl)C(O)OCi-Cs alkyl, (Co-Cs alkyl)C(O)OC2-Cs alkenyl, (Co-Cs alkyl)C(O)OC2-Cs alkynyl, (Co-Cs alkyl)C(O)OH, Co-Cs alkyl)C(O)O aryl, (Co-Cs alkyl)C(O)O heteroaryl, (Co-Cs alkyl)C(O)NR24Ci-Cs alkyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkenyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkynyl. (Co-Cs alkyl)C(O)NR24H2. (Co-Cs alkyl)C(O)NR24aryl, or (Co-Cs alkyl)C(O)NR24heteroaryl; andR24is hydrogen or C1-C7 alkyl.

[0135] For example, R1is hydrogen, propionate, acetate, benzoate, or sulfate; R5is hydrogen, ethynyl, hydroxyl; and R6is acetate, cypionate, hemisucciniate, enanthate, or propionate.

[0136] Nonlimiting examples of the compound of Formula B include 17|3-estradiol, modified forms of estradiol such as P-estradiol 17-acetate, -estradiol 17-cy pi onate, P-estradiol 17-enanthate, P-estradiol 17-valerate, P-estradiol 3,17-diacetate, P-estradiol 3,17-dipropionate, P-estradiol 3-benzoate, P-estradiol 3-benzoate 17-n-butyrate, P-estradiol 3-glycidyl ether, P-estradiol 3-methyl ether, P-estradiol 6-one, P-estradiol 3-glycidyl, P-estradiol 6-one 6-(O-carboxymethyloxime), 16-epiestriol, 17-epiestriol, 2-methoxy estradiol, 4-methoxy estradiol,32993-439275estradiol 17-phenylpropi onate, and 17| -estradiol 2-methyl ether, 17a-ethynylestradiol, megestrol acetate, estriol, and derivatives thereof. In some embodiments, carbon 17 has a ketone substituent and R5and R6are absent (e.g. estrone). Some of the aforementioned compounds of Formula B are shown below:

[0137] In embodiments wherein Y comprises a structure of Formula B, Y is conjugated to L (e.g. when L is a linking group) or Q (e.g. when L is a bond) at any position of Formula B that is capable of reacting with Q or L. One skilled in the art could readily determine the position of conjugation on Formula B and means of conjugation of Formula B to Q or L in view of general knowledge and the disclosure provided herein. In some embodiments, Formula B is conjugated to L or Q at any of positions 1, 2, 3, 4, 5, 6, 7, 8. 9, 10, 11. 12. 13. 14, 15, 16, 17, 18, 19, or 20 of Formula B. In some embodiments, Formula B is conjugated to L or Q at position 3 or 17 of Formula B.

[0138] In other embodiments. Y acts at an estrogen receptor but is not encompassed by Formula B. Nonlimiting examples of ligands that act at an estrogen receptor that are not encompassed by Formula B are show n below:32993-439275

[0139] In some embodiments, Y acts at a glucocorticoid receptor (GR). In some embodiments, Y comprises any structure that permits or promotes agonist activity at the GR, while in other embodiments Y is an antagonist of GR. In exemplary embodiments, Y comprises a structure of Formula C:Formula Cwherein R2, R3, R6, R7, R8. R9. and R10are each independently moieties that permit or promote agonist or antagonist activity upon the binding of the compound of Formula C to the GR; and each dash represents an optional double bond. In some embodiments, Formula C further comprises one or more substituents at one or more of positions 1, 2, 4, 5, 6, 7, 8, 9, 11, 12. 14. and 15 (e.g. hydroxyl or ketone at position-11).

[0140] In some embodiments, Y comprises a structure of Formula C, wherein R2is hydrogen, halo. OH, or C1-C7 alkyl;R3is hydrogen, halo, OH, or C1-C7 alky l;R6is hydrogen, Ci-Cs alkyl, C2-C8 alkenyl, C2-C8 alkynyl, heteroalkyl, (Co-Cs alkyl)aryl, (Co-C8alkyl)heteroaryl, (Co-Cs alkyl)C(O)Ci-Cs alkyl, (Co-C8alkyl)C(O)C2-C8 alkenyl, (Co-C8alkyl)C(O)C2-Cs alkynyl, (Co-Cs alkyl)C(O)H, (Co-C8alkyl)C(O)aryl, (Co-C8alkyl)C(O)heteroaryl, (Co-Cs alkyl)C(O)OCi-Cs alkyl, (Co-Cs alkyl)C(O)OC2-Cs alkenyl, (Co-Cs alkyl)C(O)OC2-Cs alkynyl, (Co-Cs alkyl)C(O)OH, Co-Cs alkyl)C(O)O aryl, (Co-Cs alky 1)C(O)O heteroaryl, (Co-Cs alkyl)C(O)NR24Ci-C8 alkyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkenyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkynyl. (Co-Cs alkyl)C(O)NR24H2. (Co-Cs alkyl)C(O)NR24aiyl, or (Co-Cs alkyl)C(O)NR24heteroaryl;R7is hydrogen, Ci-Cs alkyl, C2-Cs alkenyl, C2-Cs alkynyl. heteroalkyl, (Co-Cs alkyl)aryl, (Co-Cs alkyl)heteroaryl, (Co alkyl)C(O)Ci-Cs alkyl, (Co alkyl)C(O)C2-Cs alkenyl, (Co alkyl)C(O)C2-C8 alkynyl, (Co)C(0)aryl, (Co)C(0)heteroaiyl, (Co)C(0)OCi-Cs alkyl, (Co alkyl)C(O)OC2-C8alkenyl, (Co alkyl)C(O)OC2-Cs alkynyl, or (Co alkyl)C(O)OH;R8is hydrogen or C1-C7 alky l;R9is hydrogen or C1-C7 alkyl;32993-439275R10is hydrogen or OH; andR24is hydrogen or C1-C7 alkyl.

[0141] For example, R2is hydrogen or methyl; R3is hydrogen, fluoro, chloro, or methyl; R6is hydrogen or C(O) C1-C7 alkyl; R7is hydrogen, C(O)CHs, or C(O)CH2CHs; R8is hydrogen or methyl; R9is hydrogen or methyl; and R10is hydroxyl.

[0142] Nonlimiting examples of structures of Formula C include:32993-439275and derivatives thereof.

[0143] In embodiments wherein Y comprises a structure of Formula C, Y is conjugated to L (e g. when L is a linking group) or Q (e.g. when L is a bond) at any position of Formula C that is capable of reacting with Q or L. One skilled in the art could readily determine the position of conjugation on Formula C and means of conjugation of Formula C to Q or L in view of general knowledge and the disclosure provided herein. In some embodiments, Formula C is conjugated to L or Q at any of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 of Formula C. In some embodiments, Formula C is conjugated to L or Q at position 3, 10, 16 or 17 of Formula C.

[0144] In some embodiments, Y acts at a mineralcorticoid receptor (MR). In some embodiments, Y comprises any structure that permits or promotes agonist activity at the MR, while in other embodiments Y is an antagonist of MR. In exemplary embodiments, Ywherein R2, R3, R7and R10are each independently a moiety that permits or promotes agonist or antagonist activity upon binding of the compound of Formula D to the MR; and the dashed line indicates an optional double bond. In some embodiments, Formula D further comprises one or more substituents at one or more of positions 1, 2, 4, 5, 6, 7, 8, 11, 12, 14, 15, 16. and 17.

[0145] In some embodiments, Y comprises a structure of Formula D, wherein R2is hydrogen, halo. OH, or C1-C7 alkyl;R3is hydrogen, halo, OH, or C1-C7 alky l;32993-439275R7is hydrogen, Ci-Cs alkyl, C2-C8 alkenyl, C2 Cs alkynyl, heteroalkyl, (Co-Cs alkyl)aryl, (Co-Cs alkyl)heteroaryl, (Co alkyl)C(O)Ci-C8 alkyl, (Co alkyl)C(O)C2-Cs alkenyl, (Co alkyl)C(O)C2-C8alkynyl, (Co)C(0)aiyl, (Co)C(0)heteroaiyl, (Co)C(0)OCi-C8alkyl, (Co alkyl)C(O)OC2-C8alkenyl, (Co alkyl)C(O)OC2-C8 alkynyl, or (Co alkyl)C(O)OH;R10is hydrogen or OH; andR24is hydrogen or C1-C7 alkyl.

[0146] For example, R2is hydrogen or methyl; R?is hydrogen, fluoro, chloro, or methyl; R7is hydrogen, C(O)CHs, or C(O)CH2CHs; and R10is hydroxyl.Nonlimiting examples of compounds of Formula D include:

[0147] In embodiments wherein Y comprises a structure of Formula D, Y is conjugated to L (e.g. when L is a linking group) or Q (e.g. when L is a bond) at any position of Formula D that is capable of reacting with Q or L. One skilled in the art could readily determine the position of conjugation on Formula D and means of conjugation of Formula D to Q or L in view of general knowledge and the disclosure provided herein. In some embodiments, Formula D is conjugated to L or Q at any of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 of Formula D. In some embodiments, Formula D is conjugated to L or Q at position 3, 10, 13, or 17 of Formula D.

[0148] In some embodiments, Y acts at a progesterone receptor (PR). In some embodiments, Y comprises any structure that permits or promotes agonist activity at the PR,32993-439275while in other embodiments Y is an antagonist of PR. In exemplary embodiments, Y comprises a structure of Formula E:Formula Ewherein R2, R3, R4, and R7are each independently moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula E to the PR; and the dashed line indicates an optional double bond. In some embodiments, Formula E further comprises one or more substituents at one or more of positions 1, 2, 4, 5, 6, 7, 8, 11, 12, 14, 15, 16. and 17 (e.g. a methyl group at position 6).

[0149] In some embodiments, Y comprises a structure of Formula E, wherein R2is hydrogen, halo. OH, or C1-C7 alkyl;R3is hydrogen, halo, OH, or C1-C7 alky l;R4is hydrogen, (Co-Cs alkyl)halo, Ci-Cs alkyd, C2-C8 alkenyl, C2-18 alkynyl, heteroalkyl, (Co-Cs alkyl)aryl, (Co-C8alkyl)heteroaryl, (Co-Cs alkyl)OCi-Cs alkyl, (Co-Cs alkyl)OC2-Cs alkenyl, (Co-Cs alkyl)OC2-Cs alkynyl, (Co-Cs alkyl)OH. (Co-Cs alkyl)SH, (Co-Cs alkyl)NR24Ci-Cs alky 1, (Co-C8alkyl)NR24C2-C8alkenyl, (Co-C8alkyl)NR24C2-Cs alkynyl, (Co-C8alkyl)NR24H2, (Co-C8alkyl)C(O)Ci-Cs alkyl, (Co-C8alkyl)C(O)C2-C8alkenyl, (Co-C8alky l)C(O)C2-Cs alkynyl, (Co-C8alky 1)C(O)H, (Co-C8alky l)C(O)ary 1, (Co-Cs alkyl)C(O)heteroaryl, (Co-Cs alkyl)C(O)OCi-Cs alkyl, (Co-Cs alkyl)C(O)OC2-C8alkenyl. (Co-Cs alkyl)C(O)OC2-Cs alkynyl, (Co-Cs alkyl)C(O)OH, (Co-Cs alkyl)C(O)O aryl, (Co-Cs alkyl)C(O)O heteroaryl, (Co-C8alky l)OC(O)Ci-Cs alkyl, (Co-Cs alkyl)OC(O)C2-Cs alkenyl, (Co-Cs alkyl)OC(O)C2-Cis alkyny l, (Co-C8alky 1)C(O)NR24CI-CS alkyl, (Co-C8alkyl)C(O)NR24C2-C8alkenyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkyny l, (Co-Cs alky 1)C(O)NR24H2. (Co-Cs alkyl)C(O)NR24ar 1, (Co-Cs alkyl)C(O)NR24heteroaryl. (Co-Cs alkyl)NR24C(O)Ci-C8alkyl, (Co-Cs alky 1)NR24C(O)C2-CS alkenyl, or (Co-Cs alky 1)NR24C(O)C2-Cs alkynyl, (Co-Cs alkyl)NR24C(O)OH, (Co-Cs alkyl)OC(O)OCi-Cs alkyl, (Co-Cs alkyl)OC(O)OC2-Cs alkenyl, (Co-Cs alkyl)OC(O)OC2-Cs alkyny l, (Co-Cs alkyl)OC(O)OH, (Co-Cs alkyl)OC(O)NR24Ci-Cs alky l, (Co-Cs aJkyl)OC(O)NR24C2-Cs alkenyl, (Co-Cs alkyl)OC(O)NR24C2-Cs alkynyl, (Co-Cs alkyl)OC(O)NR24H2, (Co-Cs alkyl)NR24(O)OCi-Cs alkyl, (Co-Cs alkyl)NR24(O)OC2-Cs alkenyl, (Co-Cs alkyl)NR24(O)OC2-Cs alkyny l, or (Co-Cs alkyl)NR24(O)OH;32993-439275R7is hydrogen, Ci-Cs alkyl, C2-C8 alkenyl, C2 Cs alkynyl, heteroalkyl, (Co-Cs alkyl)aryl, (Co-Cs alkyl)heteroaryl, (Co alkyl)C(O)Ci-C8 alkyl, (Co alkyl)C(O)C2-Cs alkenyl, (Co alkyl)C(O)C2-C8alkynyl, (Co)C(0)aiyl, (Co)C(0)heteroaiyl, (Co)C(0)OCi-C8alkyl, (Co alkyl)C(O)OC2-C8alkenyl, (Co alkyl)C(O)OC2-C8 alkynyl, or (Co alkyl)C(O)OH; andR24is hydrogen or C1-C7 alkyl.

[0150] For example, R2is hydrogen or methyl; R' is hydrogen or methyl: R4is (Ci alkyl)C(O)Ci-C4 alkyl, acetate, cypionate, hemisucciniate, enanthate, or propionate; and R7is hydrogen, C(O)CH3, or C(O)CH2CH3

[0151] Nonlimiting examples of compounds of Formula E include:and derivatives thereof.

[0152] In embodiments wherein Y comprises a structure of Formula E, Y is conjugated to L (e.g. when L is a linking group) or Q (e.g. when L is a bond) at any position of Formula E that is capable of reacting with Q or L. One skilled in the art could readily determine the position of conjugation on Formula E and means of conjugation of Formula E to Q or L in view of general knowledge and the disclosure provided herein. In some embodiments, Formula E is conjugated to L or Q at any of positions 1. 2, 3, 4, 5, 6, 7, 8. 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 of Formula E. In some embodiments, Formula E is conjugated to L or Q through position 3 or 17 of Formula E.

[0153] In other embodiments, Y acts at a progesterone receptor but is not is not encompassed by Formula E. For example, Y can comprise the below structure and analogs thereof:32993-439275

[0154] In some embodiments, Y acts at an androgen receptor (AR). In some embodiments, Y comprises any structure that permits or promotes agonist activity at the AR, while in other embodiments Y is an antagonist of AR. In exemplary embodiments, Y comprises a structure of Formula F:wherein R1, when present, R2, R3and R6are each independently a moiety that permits or promotes agonist or antagonist activity upon binding of the compound of Formula F to the AR; and each dashed line represents an optional double bond, with the proviso that no more than one of the optional carbon-carbon double bond is present at position 5. In some embodiments, Formula F further comprises one or more substituents at one or more of positions 1, 2. 4, 5, 6, 7, 8, 11, 12, 14, 15, 16, and 17.

[0155] In some embodiments, Y comprises a structure of Formula E, wherein R1is hydrogen, C1-C7 alkyl; (C0-C3 alkyl)C(O)Ci-C7alkyl, (C0-C3 alkyl)C(O)aryl, or SO3H;R2is hydrogen, halo. OH, or C1-C7 alkyl;R3is hydrogen, halo. OH, or C1-C7 alkyl;R6is hydrogen, Ci-Cs alkyl, C2-C8 alkenyl, C2-C8 alkynyl, heteroalkyl, (Co-Cs alkyl)aryl, (Co-Cs alkyl)heteroaryl, (Co-Cs alkyl)C(O)Ci-Cs alkyl, (Co-Cs alkyl)C(O)C2-C8 alkenyl, (Co-Cs alkyl)C(O)C2-Cs alkynyl, (Co-Cs alkyl)C(O)H, (Co-Cs alkyl)C(O)aryl, (Co-Cs alkyl)C(O)heteroaryl, (Co-Cs alkyl)C(O)OCi-Cs alkyl, (Co-Cs alkyl)C(O)OC2-C8 alkenyl, (Co-Cs alkyl)C(O)OC2-Cs alkynyl, (Co-Cs alkyl)C(O)OH. Co-Cs alkyl)C(O)O aryl, (Co-Cs alkyl)C(O)O heteroaryl, (Co-Cs alkyl)C(O)NR24Ci-Cs alkyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkenyl, (Co-Cs alkyl)C(O)NR24C2-Cs alkynyl, (Co-Cs alkyl)C(O)NR24H2, (Co-Cs alkyl)C(O)NR24aryl, or (Co-Cs alkyl)C(O)NR24heteroaryl; andR24is hydrogen or C1-C7 alkyl.

[0156] For example, R1is hydrogen or absent; R2is hydrogen or methyl; R3is hydrogen or methyl; and R6is H or absent.

[0157] Nonlimiting examples of compounds of Formula F include:32993-439275thereof.

[0158] In embodiments wherein Y comprises a structure of Formula F, Y is conjugated to L (e.g. when L is a linking group) or Q (e.g. when L is a bond) at any position of Formula F that is capable of reacting with Q or L. One skilled in the art could readily determine the position of conjugation on Formula F and means of conjugation of Formula F to Q or L in view of general knowledge and the disclosure provided herein. In some embodiments, Formula F is conjugated to L or Q at any of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 of Formula F. In some embodiments, Formula F is conjugated to L or Q at position 3 or 17 of Formula F.

[0159] In some embodiments, Y acts at the famesoid X receptor (FXR). In some embodiments, Y comprises any structure that permits or promotes agonist activity at the FXR. while in other embodiments Y is an antagonist of FXR. In some of these embodiments, Y is a bile acid. In exemplary embodiments, Y has a structure of Formula G:Formula Gwherein each of R13, R16, and R17are independent moieties that permit or promote agonist or antagonist activity upon binding of the compound of Formula G to a FXR.

[0160] In some embodiments when Y comprises a structure of Formula M, each of R15and R16are independently hydrogen, (Co-Cs alkyljhalo, Ci-Cis alkyl, C2-C18 alkenyl, C2-C1832993-439275alkynyl, heteroalkyl, or (Co-Cs alkyljOH; and R17is OH, (Co-Cs alkyl)NH(Ci-C4 alkyljSOsH, or (Co-Cs alkyl)NH(Ci-C4 alkyljCOOH.

[0161] In accordance with one embodiment Y is a steroid selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol. In one embodiment Y is DHEA, or estradiol.Modification of the NHR Ligand (Y)

[0162] In some embodiments, the NHR ligand is derivatized or otherwise chemically modified to comprise a reactive moiety that is capable of reacting with an incretin peptide, or the linking group (L). In the embodiments described herein, Y is derivatized at any position of Y that is capable of reacting with Q or L. The position of derivatization on Y is apparent to one skilled in the art and depends on the type of NHR ligand used and the activity that is desired. For example, in embodiments wherein Y has a structure comprising a tetracyclic skeleton having three 6-membered rings joined to one 5 -membered ring or a variation thereof, Y can be derivatized at any of positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. Other positions of derivatization can be as previously described herein.

[0163] The NHR ligand can be derivatized using any agent known to one skilled in the art or described herein. For example, estradiol can be derivatized with succinic acid, succinic anhydride, benzoic acid, ethyl 2-bromoacetate, or iodoacetic acid to form the below derivatives of estradiol that are capable of conjugating to Q or L.32993-439275

[0164] Similarly, any of the aforementioned NHR ligands can be derivatized by methods known in the art. Additionally, certain derivatized ligands are commercially available and can be purchased from chemical companies such as Sigma-Aldrich.Linker (L) of the conjugate Q-L-Y

[0165] In one embodiment the linker (L) of the conjugate Q-L-Y is a bond, molecule or group of molecules that binds the NHR ligand component (Y) to the incretin peptide. In one embodiment L comprises an amino acid or polymer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-8 and q and p are independently an integer selected from the range of 0-4. In one embodiment L comprises the structure -[COCH2(OCH2CH2)k-NH]q-, wherein k is an integer selected from the range of 2-8 and q is 1. In one embodiment, L is a bond.Multimers of the conjugate Q-L-Y

[0166] In accordance with one embodiment of the present disclosure, a multimer of incretin peptides is provided wherein one or more of the incretin peptides of the multimer is conjugated to a NHR ligand. In one embodiment a conjugate comprising an in cretin peptide dimer and an NHR ligand is provided, wherein the incretin peptide dimer comprises a first and second incretin peptide linked together via a bridging linker and the nuclear hormone receptor ligand is linked to one or both of the first and second incretin peptides. In one embodiment the conjugate comprises a dimer formed between two identical incretin peptides (i.e., a homodimer) and in one embodiment the conjugate comprises a dimer formed betw een two incretin peptides that differ in structure (i.e., a heterodimer) wherein the incretin peptides are selected from any of the incretin peptides disclosed herein in combination with any NHR ligand conjugates disclosed herein to form conjugates having the formula Q-L-Y as disclosed herein, wherein the first and second incretin peptides of the respective incretin peptides are linked to one another via a bridging linker.

[0167] In one embodiment, the incretin peptide dimers of the Q-L-Y conjugates comprise a first and second incretin peptide independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 coagonist, and a Gcg / GIP / GLP-1 tri-agonist, wherein the incretin peptides are linked together via a bridging linker joining32993-439275

[0168] i) the side chain of an amino acid independently selected from positions 12, 20, 24, 31 and 40 of the respective first and second incretin peptide, or

[0169] ii) a C-terminal amide of the first incretin peptide to a C-terminal amide of the second incretin peptide; or

[0170] iii) a C-terminal amide of the first incretin peptide to the side chain of an amino acid selected from positions 12, 24, 31 and 40 of the second incretin peptide, or

[0171] iv) the side chain of the amino acid at position 12 of the first incretin peptide to the side chain of the amino acid at position 12 of the second incretin peptide, or

[0172] v) the side chain of the amino acid at position 24 of the first incretin peptide to the side chain of the amino acid at position 24 of the second incretin peptide,

[0173] further wherein a nuclear hormone receptor ligand is linked to said heterodimer or homodimer via the side chain of an amino acid of the first and / or second incretin peptide at a position not linked to the bridging linker and selected from position 12, 17, 2024, 31 or 40 of the first and / or second incretin peptide. In one embodiment the NHR ligand is linked to the first incretin peptide at a side chain selected from 28, 31 or 40, optionally at position 40, optionally via a spacer. In one embodiment the NHR ligand is linked to the first and second incretin peptide at a side chain selected from 28, 31 or 40, optionally at position 40, optionally via a spacer. In one embodiment the NHR ligand is linked to the side chain of an amino acid at position 40 of the first and / or second incretin of the incretin peptide / NHR ligand conjugate and the bridging linker joins the side chain of the amino acid at position 12 of the first incretin peptide to the side chain of the amino acid at position 12 of the second incretin peptide, or joins the side chain of the amino acid at position 24 of the first incretin peptide to the side chain of the amino acid at position 24 of the second incretin peptide.

[0174] In one embodiment, the incretin peptide / NHR ligand conjugate comprises a first and second incretin peptide independently selected from peptides comprising the sequence

[0175] i) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSK (SEQ ID NO: 37); or

[0176] ii) Y-Aib-QGTFTSDYSIXi3LDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSK (SEQ ID NO: 42), or

[0177] iii) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPX4o (SEQ ID NO: 21); or

[0178] iv) H-Aib-EGTFTSDVSSYLEGQAAKEFIAWLVRGRGX4o (SEQ ID NO: 22); or32993-439275

[0179] v) H-Aib-EGTFTSDVSX12YLEX16QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;X13 is alpha-methyl leucine;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys, Om, Cys, or the amino acid at X40 is amidated and linked to -[COCH2(OCH2CH2)k-NH]qwherein k is an integer selected from 1-8 and q is 1 or 2;

[0180] further wherein the first and second incretin peptides are linked together via a bridging linker joining the respective the side chains of amino acids independently selected from position 12, 24, 31 or 40, optionally at position 12 or 24 of the first and second incretin peptides, and

[0181] said NHR ligand is linked to the side chain of an amino acid at a position not linked to the bridging linker and independently located at position 12, 24 or 40 of said first and / or second incretin peptide, or at a C-terminal primary amine of said first and / or second incretin peptide, optionally at the side chain of an amino acid at position 40,

[0182] optionally wherein the nuclear hormone receptor ligand is linked to a side chain of an amino acid of the first and / or second incretin peptide via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-8 and q is an integer selected from the range of 0-4, optionally wherein z is 0, k is an integer selected from the range of 2-8 and q is 0.

[0183] In one embodiment the first and second incretins of the incretin peptide / NHR ligand conjugate are identical and the incretin dimer of the of the incretin peptide / NHR ligand conjugate is a homodimer. In one embodiment the first and second incretins of the incretin peptide / NHR ligand conjugate differ from one another (thus representing a heterodimer of incretin peptides), but have the same activity at incretin receptors (e.g.. both the first and second incretin peptides are GIP / GLP-1 co-agonists). In one embodiment the first and second incretins of the incretin peptide / NHR ligand conjugate differ from one another (thus representing a heterodimer of incretin peptides), and have different activities at incretin receptors (e.g., the first incretin peptides is GIP / GLP-1 co-agonists and the second incretin peptide is a Gcg / GIP / GLP-1 tri-agonist).32993-439275

[0184] In one embodiment, the incretin peptide / NHR ligand conjugate comprises a first incretin peptide comprising the amino acid sequence:

[0185] H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys or Cys; and

[0186] a second incretin peptide comprising the amino acid sequenceH-Aib-EGTFTSDVSXio2YLEXio6QAAKEFIX2o4WLVX2o8GRPSSGAPPPSX4oo (SEQ ID NO: 24), whereinX102 is Ser, Lys or Cys;X106 is Gly or Glu;X2o4 is Ala, Lys or Cys;X208 is Arg or Lys; andX400 is Lys or Cys, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 or 24 of said first incretin peptide to the side chain of an amino acid at position 12 or 24 of said second incretin peptide, and said NHR ligand is linked to the side chain of the amino acid at X40 and / or X400, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4, optionally wherein z is 0, k is an integer selected from the range of 2-8 and q is 0.

[0187] In any of the incretin peptide / NHR ligand conjugates disclosed herein the NHR ligand can be linked to incretin peptide or incretin peptide multimer via a direct bond (i.e.L of Q-L-Y is a bond).

[0188] In accordance with one embodiment the bridging linking joining the first and second incretin of the incretin peptide / NHR ligand conjugate comprises a structure selected from:i) a disulfide bond;ii) -NH(CO)(CH2)n-S-, wherein n is 1-4; oriii) a triazole.32993-439275

[0189] In one embodiment, the bridging linker comprises a disulfide bond formed between the side chains of cysteine residues amino acid positions of the respective first and second incretin peptides independently selected from positions 12, 24, 28 and 40, optionally wherein cysteine residues are at the same respective position of each of the first and second incretin peptides. In one embodiment a cysteine residue is present at position 12 of each of the first and second incretin peptides. In one embodiment a cysteine residue is present at position 24 of each of the first and second incretin peptides.

[0190] In one embodiment, the bridging linker comprises a thioether bond formed between the side chains of a cysteine and a lysine amino acid in the respective first and second incretin peptides, wherein the cysteine and lysine residues are located at positions independently selected from positions 12, 24, 28 and 40 of the respective first and a second incretin peptides. In one embodiment a cysteine is present at a position selected from 12, 24, 28 and 40 of the first incretin peptide, and the side chain of the cysteine is linked to the side chain of a lysine present at a position selected from 12, 24, 28 and 40 of the second incretin peptide via the linking moiety -NH(CO)(CH2)n-S-, wherein n is 1-4, optionally wherein n is 2. In one embodiment the bridging linker joins the side chains of a cysteine and lysine via linking moiety -NH(CO)(CH2)n-S-, wherein n is 1-4, and the linked cysteine and lysine residues are located at the same respective position on the first and second incretin peptides. In one embodiment the side chain of a cysteine residue present at position 12 of the first incretin peptide and is linked to the side chain of a lysine residue present at position 12 of the second incretin peptide. In one embodiment the side chain of a cysteine at position 24 is linked to the side chain of the lysine at position 24 of the second incretin peptide via the linking moiety -NH(CO)(CH2)n-S-, wherein n is 1-4, optionally wherein n is 2.

[0191] In one embodiment click chemistry is used to link the first and second incretin peptides wherein the bridging linker comprises a triazole. Specifically, first and second incretin peptides are provided wherein the side chain of an amino acid of the first incretin peptide comprises an alkyne group and the side chain of an amino acid of the second incretin peptide comprises azide, wherein the alkyne group reacts with the azide group to from a triazole joining the first and second peptides. In one embodiment the side chain of an amino acid at a position selected from positions 12, 24, 38 and 40 of the first incretin peptide is modified to comprise an azide group, and an amino acid at a position selected from positions 12, 24, 38 and 40 of the second incretin peptide is modified to comprise an alkyne group, and a bridging link is formed between the two respective modified amino acids to join the first and second incretin peptides via a bridging linker comprising a triazole. In one embodiment the bridging linker comprising a triazole joins the side chains of amino acids present at position 12 of the32993-439275respective first and second incretin peptide. In one embodiment the bridging linker comprising a triazole joins the side chains of amino acids present at position 24 of the respective first and second incretin peptide.

[0192] In one embodiment, the incretin peptide / NHR ligand conjugate dimer comprises a heterodimer or homodimer of two incretin peptides wherein a first incretin peptide comprising the sequence of

[0193] H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX2sGRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Cys;Xi6 is Gly or Glu;X24 is Ala;X28 is Lys; andX40 is Lys or Cys; anda second incretin peptide comprising the sequenceH-Aib-EGTFTSDVSXi02YLEXi06QAAKEFIX204WLVX208GRPSSGAPPPSX400 (SEQ ID NO: 24), whereinX102 is Lys;X106 is Gly or Glu;X204 is Ala;X208 is Lys; andX400 is Lys or Cys, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 of said first incretin peptide to the side chain of an amino acid at position 12 of said second incretin peptide, optionally via a linking moiety comprising the structure - NH(CO)(CH2)n-S-, wherein n is 1-4; or via a linking moiety comprising a triazole, whereinX40 is absent, Lys or an amino acid comprising an NHR ligand linked to its side chain, optionally via a first spacer; andX400 is an amino acid comprising an NHR ligand linked to its side chain, optionally via a second spacer,wherein said first and second spacer is independently selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.32993-439275

[0194] In one embodiment, the incretin peptide / NHR ligand conjugate dimer comprises a heterodimer or homodimer of two incretin peptides independently selected from SEQ ID NOs: 19-24 wherein X40and X400are each Lys having a NHR ligand linked to the side chain of said Lys, wherein:

[0195] i) the NHR ligand linked to the Lys of X40 and X400 is independently selected from a PPAR agonist, a famesoid X receptor (FXR) agonist such as cilofexor, or a steroid, optionally wherein the PPAR agonist is a compound selected from the group consisting of GW501 16, Pemafibrate, Chiglitazar and Saroglitzazar, and the steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;

[0196] ii) the NHR ligand linked to Lys of X40 and X4oois the same, optionally wherein the NHR ligand is a famesoid X receptor (FXR) agonist such as cilofexor, a PPAR agonist selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, or a steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;

[0197] iii) the NHR ligand linked to Lys of X40 is a PPAR agonist and the NHR ligand linked to Lys of X400 is a steroid, optionally wherein the PPAR agonist is selected from the group consisting of GW501516. Pemafibrate, Chiglitazar and Saroglitzazar, and the steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;

[0198] iv) the NHR ligand linked to Lys of X40 is GW501516, and the NHR ligand linked to Lys of X400 is DHEA;

[0199] v) the NHR ligand linked to Lys of X40 and X400 is GW501516; or

[0200] vi) the NHR ligand linked to Lys of X40 and X400 is DHEA.

[0201] In one embodiment, the incretin peptide / NHR ligand conjugate dimer comprises a heterodimer or homodimer of two incretin peptides wherein said first incretin comprises the sequence of

[0202] H-Aib-EGTFTSDVSCYLEEQAAKEFIAWLVKGRPSSGAPPPSK (SEQ ID NO: 46); and

[0203] the second incretin peptide comprises the sequence

[0204] H-Aib-EGTFTSDVSKYLEEQAAKEFIKWLVX2sGRPSSGAPPPSK (SEQ ID NO: 47); wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 of said first incretin peptide to the side chain of an amino acid at position 12 of said second incretin peptide via a linker comprising the the structure:

[0205] -NH(CO)(CH2)2-S-, and a NHR ligand is linked to the side chain of an amino acid at position 40 of the first and / or second incretin peptide, wherein the nuclear hormone32993-439275receptor ligand is independently selected from the group consisting of DHEA, estrogen, dexamethasone, GW501516, Pemafibrate, Chiglitazar and Saroglitzazar.

[0206] In one embodiment a conjugate is provided comprising a multimer of incretin peptides incretin peptides linked to one another to form dimers, trimer and tetramers of incretin peptides wherein one or more of the incretin peptides forming the incretin dimer, trimer or tetramers comprises a NHR ligand covalently linked to the side chain of an amino acid selected from positions 24, 28, 31, 40 or any amino acid of a C-terminal extension of the incretin peptide.

[0207] In one embodiment, a conjugate is provided comprising a tetramer of incretin peptides wherein each of the four incretin peptides are independently selected from SEQ ID NO: 19-24, wherein the first and second incretin peptides are linked to form a first dimer and the third and fourth incretin peptides are linked to form a second dimer, wherein the incretin peptides of the respective first and second dimers are linked via a linker comprising the structure: -[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]s wherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2, and a tetramer linker joins the first and second dimers to form a tetramer wherein the tetramer linker comprises the structure -(CH2)m(CHCONH2)-, where m is an integer selected form 1 to 4, that is linked to the primary amine of each of the respective bridging linkers joining the first incretin peptide to the second incretin peptide and the third incretin peptide to the fourth incretin peptide.

[0208] In one embodiment an incretin peptide / NHR ligand multimer is provided comprising a dimer of any of the incretin peptide / NHR ligand conjugates disclosed herein. In one embodiment the multimer comprises a first and second incretin peptide linked to one another to form an incretin peptide heterodimer or homodimer wherein the first incretin peptide comprises the sequence ofH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFlX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys; andthe second incretin peptide comprising the sequenceH-Aib-EGTFTSDVSXio2YLEXio6QAAKEFIX2o4WLVX2o8GRPSSGAPPPSX4oo (SEQ ID NO: 24), wherein32993-439275X102 is Ser, Lys or Cys;X106 is Gly or Glu;X204 is Ala. Lys or Cys;X208 is Arg or Lys; andX400 is Lys, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chains of the Lys at position 40 of the respective first and second incretin peptides, said bridging linker comprising the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2; further whereina nuclear hormone receptor ligand is linked to the side chain of an amino acid of the first incretin peptide and / or the second incretin peptide at position independently selected from positions 12, 20, and 24, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4,wherein an incretin peptide, an incretin peptide / NHR ligand conjugate, or an incretin peptide / NHR ligand conjugate dimer is linked to said bridging linker via the amide group of the bridging linker to form a tetramer of incretin peptides.

[0209] In one embodiment, the incretin peptide / NHR ligand conjugate multimer comprises a tetramer of four incretin peptides, wherein the first, second, third and fourth incretin peptides are independently selected from the sequence H-Aib-EGTFTSDVSX12YLEX16QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys;wherein the first and second incretin peptides are linked to one another via a first bridging linker joining the side chains of the Lys at position 40 of the respective first and second incretin peptides to form a first dimer, and the third and fourth incretin peptides are linked to one another via a second bridging linker joining the side chains of the Lys at position 40 of the respective third and fourth incretin peptides to form a second dimer,32993-439275said first and second bridging linkers independently comprising the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2; and

[0210] a NHR ligand is linked to the side chain of an amino acid of one or more of the first, second, third or fourth incretin peptide at a position independently selected from positions 12, 20, and 24, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4, wherein the first dimer is linked to the second dimer via a tetramer linker comprises the structure -(CH2)m(CHCONH2)-, where m is an integer selected form 1 to 4, that is linked to the primary amine of each of the respective bridging linkers joining the first incretin peptide to the second incretin peptide and the third incretin peptide to the fourth incretin peptide.Prodrug Derivatives

[0211] The present disclosure also encompasses prodrug derivative of the incretin peptide / NHR ligand conjugates disclosed herein. In one embodiment a prodrug derivative of any of the incretin peptide / NHR ligand conjugates, or multimers thereof, disclosed herein is provided wherein a dipeptide prodrug element is covalently linked to an active site of the bioactive polypeptide via an amide linkage. The amino acids, A and B of the dipeptide prodrug element are selected to allow the dipeptide prodrug element to chemically cleave under physiological conductions to release the dipeptide (forming a diketopiperazine) and the active the incretin peptide / NHR ligand conjugate. In one embodiment a dipeptide prodrug element is covalently linked to one or more amino acids at positions 1, 17 or 16 of one or more of the incretin peptides of the incretin peptide / NHR ligand conjugate. In some embodiments the dipeptide prodrug element is linked to the N-terminal alpha amine of incretin peptide / NHR ligand conjugate or at one or more of the incretin peptides of the incretin peptide / NHR ligand conjugate multimers disclosed herein. Subsequent removal of the dipeptide, under physiological conditions and in the absence of enzy matic activity7, restores full activity7to the incretin peptides of the incretin peptide / NHR ligand conjugate peptide.

[0212] In accordance with one embodiment the peptide of SEQ ID NO: 16 is provided wherein X? is isoacyl-Thr, and Xr> is alpha methylated amino acid, further wherein a dipeptide prodrug element is linked via an amide bond to the alpha amine of the isoacyl-Thr of the Gcg / GLP-l / GIP tri-agonist peptide of SEQ ID NO: 16. In accordance with one embodiment the peptide of SEQ ID NO: 16 is provided wherein X20 is Lys, wherein a dipeptide prodrug element is linked via an amide bond to the alpha amine of the Lys side chain.32993-439275

[0213] In some embodiments a dipeptide prodrug element is provided having the general structure of A-B wherein A is an amino acid and B is an -alky I at cd amino acid linked to the incretin peptide / NHR ligand conjugate through formation of an amide bond between a carboxyl of B (in A-B) and an amine of the the incretin peptide / NHR ligand conjugate peptide. Furthermore, in some embodiments, the chemical cleavage half-life of A-B from the incretin peptide / NHR ligand conjugate is at least about 1 week in PBS under physiological conditions, and in one embodiment is the chemical cleavage half-life of A-B from the incretin peptide / NHR ligand conjugate is greater than 14 days. In one embodiment the chemical cleavage half-life of A-B from the incretin peptide / NHR ligand conjugate is at least about 8 days up to 21 days or is about 15 days.

[0214] In some embodiments A and / or B is an amino acid in the D stereoisomer configuration. In some exemplary embodiments, A is an amino acid in the D stereoisomer configuration and B is an ammo acid in the L stereoisomer configuration. In some exemplary embodiments, A is an amino acid in the L stereoisomer configuration and B is an amino acid in the D stereoisomer configuration. In some exemplary' embodiments, A is an amino acid in the D stereoisomer configuration and B is an amino acid in the D stereoisomer configuration.

[0215] As disclosed herein prodrugs of the incretin peptide / NHR ligand conjugates disclosed herein can be administered as highly effective therapeutics for lowering body weight in obese rodents. These peptides demonstrate a much-sustained duration of pharmacological action that should serve to minimize frequency in human dosing. They also demonstrate a much-reduced peak to trough concentration between injections which supports more precise dosing to achieve maximal efficacy (weight lowering) and minimal toxicity (nausea, flatulence and vomiting). The chemical form of the novel peptides is additionally supportive of non-injectable forms of administration, most notably by oral and inhalation routes of administration. However, any of the standard routes of administration, including intramuscular injection, can be used to administer the incretin peptide / NHR ligand conjugates to a patient.

[0216] Peptides having glucagon, GIP and GLP-1 receptor agonist activity are particularly advantageous for inducing weight loss or preventing weight gain, as well as for treating metabolic syndrome, hyperglycemia, and diabetes. Thus, in one aspect, the invention provides methods for inducing weight loss or preventing weight gain, as well as methods of treating diabetes and metabolic syndrome, which involve administering to a patient in need thereof an effective amount of an incretin peptide / NHR ligand conjugate or multimer thereof, that exhibits activity at the glucagon, GIP and the GLP-1 receptors.

[0217] In accordance with one embodiment a prodrug derivative of any of the incretin peptide / NHR ligand conjugates disclosed herein is provided. In one embodiment one32993-439275or more of the incretin peptides of the incretin peptide / NHR ligand conjugates are modified by the covalent linkage of one or more dipeptides (A-B) to an aliphatic primary7amine of the incretin peptide, wherein A is an amino acid and B is an N-alkylated amino acid linked to said peptide through an amide bond between a carboxyl moiety of B and an amine of the peptide, optionally wherein the side chain of the first amino acid (A) of said dipeptide is acylated with a C16-C30 fatty7acid, a C16-C30 phosphonic fatty7acid, or a C16-C30 diacid. In accordance with one embodiment a dipeptide prodrug element is covalently linked to an amino acid at position 1, 7 or 16 of the incretin protein. In one embodiment a dipeptide prodrug element is covalently7linked to the N-terminal alpha amine of one or both of the first and second incretin peptides of an incretin peptide / NHR ligand conjugate dimer disclosed herein.

[0218] In one embodiment, the dipeptide linked to the amino acid at position 1 is linked to the N-terminal alpha amine of any of the incretin peptide / NHR ligand conjugates disclosed herein. In one embodiment, the dipeptide is linked to the amino acid at position 7, wherein position 7 comprises an isoacyl-Thr.

[0219] In accordance with one embodiment the dipeptide prodrug element comprises the structure:wherein

[0220] (I) Ri and R2 are independently selected from the group consisting of H, Ci-Cis alkyl. C2-C18 alkenyl, (Ci-Cis alkyljOH, (Ci-Cis alkyljSH, (C2-C3 alkyl)SCH3, (C1-C4 alkyl)CONH2, (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH2, (C1-C4 alkyl)NHC(NH2+)NH2, (C0-C4 alkyl)(Cs-C6 cycloalky l), (C0-C4 alkyl)(C2-Cs heterocyclic), (C0-C4 alkyl)(C6-Cio aryl)R7, (Ci-C4 aJkyl)(C3-C9 heteroary l), and C1-C12 alkyl(Wi)Ci-Ci2alkyl, wherein Wi is a heteroatom selected from the group consisting of N, S and O, or Ri and R2together with the atoms to which they are attached form a Cs-Ci2cycloalkyl;R3 is CDs, or Ci-Cis alkyd;R4 and Rs are independently H or D;R5 is NHRe, or R5 and R2together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;Re is H or C1-C4 alkyl; and.32993-439275R? is selected from the group consisting of H, OH, halo, (C1-C7 alkyl), (C2-C7 alkenyl), OCF3, NO2, CN, NC, O(CI-C7 alky l), CO2H, CO2(Ci-C7 alkyl), NHRe, aryl, and heteroaryl, with the proviso that when A-B is linked to the alpha amino group on the N-terminus of Q and (i) both Ri and R2 are H. and (ii) R3 is methyl, then Q is not F7GLP- 1(8-37);

[0221] (II) Ri and R2 are independently selected from the group consisting of H, Ci-Ci8 alkyl, C2-C18 alkenyl, (Ci-Cis alkyl)OH, (C1-C18 alkyl)SH, (C2-C3 alkyl)SCH3, (C1-C4 alkyl)CONH2, (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH2, (C1-C4 alkyl)NHC(NH2+)NH2, (C0-C4 alkyl)(C3-C6 cycloalkyl), (C0-C4 alkyl)(C2-C5heterocyclic), (C0-C4 alkyl)(C6-Cio aryl)R7, (Ci-C4 alkyl)(C3-C9 heteroaryl), and Ci-C 12 alkyl(Wi)Ci-Ci2 alkyl, wherein Wi is a heteroatom selected from the group consisting of N, S and O, or Ri and R2 together with the atoms to which they are attached form a C3-C12 cycloalkyl;Rs is CDs or Ci-Cis alkyl;R4 is selected from the group consisting of D, CH3, CH2(CI-CIO alkyl), CH2(C2-C10 alkenyl), CH2(CO-CIO alkyl)OH, CH2(CO-CIO alkyl)SH, CH2(Co-C3alkyl)SCH3, CH2(Co-C3 alkyl)CONH2, CH2(Co-C3 alkyl)COOH, CH2(Co-C3 aJkyl)NH2, CH2(Co-C3 alkyl)NHC(NH2+)NH2, CH2(Co-C3alkyl)(C3-C6cycloalkyl), CH2(Co-C3 alkyl)(C2-C5heterocyclic), CH2(Co-C3 alkyl)(C6-Cio aryl)R?. CH2(Ci-C3 alkyl)(Cs-C9 heteroaryl), and CH2(Co-Ci2 alkyl)(Wi)Ci-Ci2 alkyl, wherein Wi is a heteroatom selected from the group consisting of N, S and O, or R4 and Rs together with the atoms to which they are attached form a pyrrolidine, a 3,4-dehydropyrrolidine, a hydroxypyrrolidine, a piperdine or a hydroxypiperdine ring;Rs is H or D;R5 is NHRe, or R5 and R2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;Re is H or C1-C4 alkyl; and,R7 is selected from the group consisting of H, OH, halo, (C1-C7 alkyl), (C2-C7 alkenyl), OCF3. NO2, CN, NC, O(Ci-C7alkyl), CO2H, CO2(Ci-C7 alkyl), NHRe, aryl, and heteroaryl;

[0222] (III) Ri and R2are independently selected from the group consisting of H. Ci-Cis alkyl, C2-C18 alkenyl, (Ci-Cis alkyl)OH, (Ci-Cis alkyl)SH, (C2-C3 alkyl)SCH3, (C1-C4 aJkyl)CONH2, (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH2, (C1-C4 alkyl)NHC(NH2+)NH2, (C0-C4 alkyl)(C3-Ce cycloalkyl), (C0-C4 alkyl)(C2-C5heterocyclic), (C0-C4 alkyl)(C6-Cio aryl)R7, (Ci-C4 alkyl)(C3-C9 heteroaryl), and C1-C12 alkyl(Wi)Ci-Ci2 alkyl, wherein Wi is a heteroatom selected from the group consisting of N, S and O, or Ri and R2 together with the atoms to which32993-439275they are attached form a C3-C12 cy cloalk l: or Ri and R2 together with the atoms to which they are attached form a C3-C12 cycloalky l;R3 is Ci-Cis alkyl;R4 is independently selected from the group consisting of CH(Ci-Cs alkyl)2, CH (C2-C8 alkenyl)2, CH(CI-C8alkyl)(OH), CH(Ci-Cs alkyl)((Ci-Cs alkyl)SH), and CH(Ci- C3 alkyl)((Ci-C8 alkyl)(NH2j), or R4 and R3 together with the atoms to which they are attached form a pyrrolidine, a 3,4-dehydropyrrolidine, a hydroxypyrrolidine, a piperdine or a hydroxypiperdine ring;Rs is H;R5 is NHRe, or R5 and R2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;Re is H or C1-C4 alkyl; and,R7 is selected from the group consisting of H, OH, halo, (C1-C7 alkyl), (C2-C7 alkenyl), OCF3. NO2, CN, NC, O(Ci-C7alkyd), CO2H, CO2(Ci-C7 alkyl), NHRe, aryl, and heteroaryd;

[0223] (IV) Ri and R2 are independently selected from the group consisting of H, C1-C18 alkyl, C2-C18 alkenyl, (C1-C18 alky 1)OH, (Ci-Cis alkyl)SH, (C2-C3 alkyl)SCH3, (C1-C4 alkyd)CONH2, (C1-C4 alkyl)COOH, (C1-C4 alky l)NH2, (C1-C4 alky 1)NHC(NH2+)NH2, (C0-C4 alkyl)(C3-Ce cycloalkyl), (C0-C4 alkyl)(C2-Cs heterocyclic), (C0-C4 alkyl)(C6-Cio aryl)R7, (Ci-C4 alkyl)(C3-C9 heteroaryl), and C1-C12 alkyl(Wi)Ci-Ci2 alkyl, wherein Wi is a heteroatom selected from the group consisting of N, S and O, or Ri and R2 together with the atoms to which they are attached form a C3-C12 cycloalkyd;R3 is CD3 or Ci-Cis alkyd;R4 and Rs are independently D or H;R5 is NHRe, or R5 and R2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;Re is H or C1-C4 alkyl; and,R7 is selected from the group consisting of H, OH, halo, (C1-C7 alkyl), (C2-C7 alkenyl), OCF, NO2. CN, NC. O(Ci-C7alkyd), CO2H, CO2(Ci-C7 alkyl), NHRe, aryl, and heteroaryl;

[0224] (V) Ri and R2 are independently selected from the group consisting of H, Ci-Cis alkyd, C2-C18 alkenyl, (C1-C18 alkyl)OH, (Ci-Cis alkyl)SH, (C2-C3 alkyl)SCH3, (C1-C4 alkyl)CONH2, (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH2, (C1-C4 alkyl)NHC(NH2+)NH2, (C0-C4 alkyl)(C3-Ce cycloalky l), (C0-C4 alkyl)(C2-Cs heterocyclic), (C0-C4 alkyl)(Ce-Cio aryl)R?, (Ci-32993-439275C4 alkyl)(C3-C9 heteroaryl), and Ci-C 12 alkyl(Wi)Ci-Ci2 alky l, wherein Wi is a heteroatom selected from the group consisting of N, S and O, or Ri and R2 together with the atoms to which they are attached form a C3-C12 cycloalkyl; or Ri and R2 together with the atoms to which they are attached form a C3-C12 cycloalkyl:R3 is Ci-Cis alkyl;R4 is independently selected from the group consisting of CH(Ci-Cs alkyl)2, CH (C2-C8 alkenyl)2, CH(CI-C alkyl)(OH), CH(CI-C alkyl)((Ci-Cs alkyl)SH), and CH(Ci- C3 alkyl)((Ci-Cs alkyd)(NH2>) or R4 and R3 together with the atoms to which they are attached form a pyrrolidine, a 3,4-dehydropyrrolidine, a hydroxypyrrolidine, a piperdine or a hydroxypiperdine ring;Rs is H;R5 is NHRe, or Rs and R2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;Rs is H or C1-C4 alkyl; and,R7 is selected from the group consisting of H, OH, halo, (C1-C7 alkyl), (C2-C7 alkenyl), OCF3, NO2. CN, NC, O(Ci-C7alkyl), CO2H, CO2(Ci-C7alkyl), NHR6, aryl, and heteroaryl; or

[0225] (VI) Ri and R2 are independently selected from the group consisting of H, Ci-Cis alkyl, C2-C18 alkenyl, (Ci-Cis alkyl)OH, (C1-C18 alkyl)SH, (C2-C3 alkyl)SCH3, (C1-C4 alkyl)CONH2, (C1-C4 alkyl)COOH. (C1-C4 alkyl)NH2, (C1-C4 alkyl)NHC(NH2+)NH2, (C0-C4 alkyl)(C3-Cs cycloalkyl), (C0-C4 alkyl)(C2-Cs heterocyclic), (C0-C4 alkyl)(Cs-Cio aryl)R7, (Ci-C4 alkyl)(C3-C9 heteroaryl), and Ci-C 12 alkyl(Wi)Ci-Ci2 alky l, wherein Wi is a heteroatom selected from the group consisting of N, S and O, or Ri and R2 together with the atoms to which they are attached form a C3-C12 cycloalkyl; or Ri and R2 together with the atoms to which they are attached form a C3-C12 cycloalkyl;R3 is CDs or Ci-Cis alkyl;R4 is independently selected from the group consisting of D, CH(Ci-Cs alkyl)2, CH (C2-Cs alkenyl)2, CH(Ci-Cs alkyl)(OH), CH(Ci-Cs alkyl)((Ci-Cs alkyl)SH), and CH(Ci-Cs alkyl)((Ci-C8 alkyl)(NH2)) or R4 and Rs together with the atoms to which they are attached form a pyrrolidine, a 3,4-dehydropyrrolidine, a hydroxypyrrolidine, a piperdine or a hydroxypiperdine ring;Rs is H or D;R5 is NHRs, or Rs and R2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;32993-439275Re is H or C1-C4 alkyl; and,R7 is selected from the group consisting of H, OH, halo, (C1-C7 alkyl), (C2-C7 alkenyl), OCF3, NO2. CN, NC, O(Ci-C7alkyl), CO2H, CO2(Ci-C7alkyl), NHR6, aryl, and heteroarvl.

[0226] (VII) Ri is (C1-C4 alkyl)NH2, optionally ILys or dLys, acylated with a C16-C20 fatty acid, a C16-C20 phosphonic fatty acid, or a C16-C20 diacid;R2 and Rs are both H;R4 and R3 together with the atoms to which they are attached form a pyrrolidine, a 3,4-dehydropyrrolidine, a hydroxypyrrolidine, a piperdine or a hydroxypiperdine ring; and Rs is NH2., optionally wherein R4 and R3 together with the atoms to which they are attached form a hydroxy substituted pyrrolidine ring or a piperdine ring.

[0227] (VIII) Ri is (C1-C4 alkyl)NH2, optionally ILys or dLys, acylated with a C16-C20 fatty acid, a C16-C20 phosphonic fatty acid, or a C16-C20 diacid;R2 is H;R4 and Rs are independently H or D;Rs is CD? or C1-C4 alkyl; andR5 is NH2, optionally wherein R4 and Rs are each D, and R3 is CD3.

[0228] IX) Ri is (C1-C4 alkyl)NH2, optionally ILys or dLys, acylated with a C16-C20 fatty acid, a C16-C20 phosphonic fatty' acid, or a C16-C20 diacid;R2, R4 and Rs are each H;R3 is CH3; andR5is NH2.

[0229] In accordance with one embodiment a prodrug derivative of any of the incretin peptide / NHR ligand conjugates disclosed herein is provided, wherein a dipeptide is covalently linked via an amide bond to the N-termmal alpha amine of one or both of the incretin peptides comprising the incretin peptide / NHR ligand conjugate, wherein the dipeptide comprises the structure:wherein32993-439275

[0230] Ri comprises a side chain of (C1-C4 alkyl)NH2, wherein a C16-C30 fatty acid, a C16-C30 phosphonic fatty7acid, or a C16-C30 diacid is covalently linked to said side chain, optionally via a spacer;

[0231] R2, is H, or C1-C4 alkyl;

[0232] R4is H, D, C1-C4 alkyl, C1-C4 alkenyl, -(CH2)nCH2OH or (C1-C4 alkyl)phenyl, wherein n is an integer selected from 0-2;

[0233] R3 is CD3, C1-C6 alkyl, or R3 and R4 together with the atoms to which they are attached form a pyrrolidine, a 3,4-dehydropyrrolidine, a hy droxypyrrolidine, a piperdine or a hydroxypiperdine ring;

[0234] R8, is H, D or C1-C4 alkyl; and

[0235] Rs is NH2, wherein said spacer is selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

[0236] In one embodiment the dipeptide comprises the structure:wherein

[0237] Ri comprises a side chain of (C4 alkyl)NH2, having a C18-C20 fatty acid, aC18-C20 phosphonic fatty7acid, or a C18-C20 diacid covalently linked to said side chain, optionally via said spacer;

[0238] R2is H;

[0239] R4 and Rs are both D or H;

[0240] R3 is CD3 or Ci-Ce alkyl; and

[0241] Rs is NH2, wherein said spacer comprises -[COCH2(OCH2CH2)k-NH]q- (gamma glutamic acid)P, wherein k is 2, q is 1 or 2 and p is 0 or 1, optionally with the proviso that when R4 and Rs are both H, Rs is CDs, optionally wherein R4 and Rs are both D and R3 is CD3. In a further embodiment the spacer is -[COCH2(OCH2CH2)2-NH]-(gamma glutamic acid)-, and the acyl group is a C18-C20 diacid.

[0242] In one embodiment the dipeptide comprises the structure:32993-439275whereinRi comprises a side chain of (C4 alkyl)NH2, having a C18-C20 fatty acid, a C18-C20 phosphonic fatty acid, or a C18-C20 diacid covalently linked to said side chain, optionally via said spacer;R2 is H;R4 and Rs are both D;R3 is CDs; andRs is NH2, wherein said Pl spacer comprises -[COCH2(OCH2CH2)k- NH]q-(gamma glutamic acid)P, wherein k is 2, q is 1 or 2 and p is 0 or 1. In a further embodiment the spacer is -[COCH2(OCH2CH2)2-NH]-(gamma glutamic acid)-, and the acyl group is a C18-C20 diacid..

[0243] In accordance with one embodiment, in any of the prodrug derivative disclosed herein, the first amino acid of the dipeptide prodrug element is an amino acid in the D-configuration, optionally wherein the amino acid is dLys.

[0244] In accordance with one embodiment a prodrug derivative of any of the incretin peptide / NHR ligand conjugates is provided wherein a sequential dipeptide structure of

[0245] is covalently linked via an amide bond to one or more of the incretin peptides comprising the incretin peptide / NHR ligand conjugates, optionally at one or more of amino acid positions 1, 12 and 17. wherein

[0246] Ri is selected from an acylated amino acid having a side chain comprising (C1-C4 alkyl)NH2 covalently linked to a Cl 6-C30 fatty acid, a Cl 6-C30 phosphonic fatty acid, or a C16-C30 diacid, optionally via a DD1 spacer;

[0247] R21 is selected from an acylated amino acid having a side chain comprising (C1-C4 alkyl)NH2covalently linked to a C16-C30 fatty acid, a C16-C30 phosphonic fatty acid, or a C16-C30 diacid, optionally via a DD2 spacer;

[0248] R2, and R22are independently H, or C1-C4 alkyl;32993-439275

[0249] Ri. RS, R24 and R28 are independently H, D or C1-C4 alkyl;

[0250] R3 and R23 are independently CD3, Ci-Cs alkyl, or R3 and R4 together with the atoms to which they are attached form a substituted or unsubstituted pyrrolidine, hydroxypyndine or piperdine nng, wherein said DD1 and DD2 spacers are independently selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4. In one embodiment the sequential dipeptide is linked at the N-terminal alpha amine of one or more of the incretin peptide comprising any of the incretin peptide / NHR ligand conjugates disclosed herein, optionally with a dipeptide having the structure of

[0251] covalently linked to the N-terminal alpha amine of one or more of the incretin peptides of the incretin peptide / NHR ligand conjugates wherein

[0252] Ri comprises a side chain of (C1-C4 alkyl)NH2, wherein a C 16-C30 fatty acid, a C16-C30 phosphonic fatty acid, or a C16-C30 diacid is covalently linked to said side chain, optionally via a Pl spacer;

[0253] R2, is H, or C1-C4 alkyl;

[0254] R4is H, D, C1-C4 alkyl, C1-C4 alkenyl, -(CH2)nCH2OH or (C1-C4 alkyl [phenyl, wherein n is an integer selected from 0-2;

[0255] R3 is CDs, Ci-Ce alkyl, or R3 and R4 together with the atoms to which they are attached form a pyrrolidine, a 3,4-dehydropyrrolidine, a hydroxypyrrolidine, a piperdine or a hydroxypiperdine ring;

[0256] Rs, is H, D or C1-C4 alkyl; and

[0257] Rs is NH2, wherein said Pl spacer is selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

[0258] Acylation and alkylation

[0259] In one embodiment the incretin peptide / NHR ligand conjugates of the present invention are acylated or akylated at position 17 and / or at the first ammo acid of the32993-439275dipeptide prodrug element of the prodrug derivative of the incretin peptide / NHR ligand conjugates. In one embodiment the incretin peptide / NHR ligand conjugates of the present invention are acylated or akylated at N-terminal alpha amine of the incretin peptide. In one embodiment a incretin peptide / NHR ligand conjugate dimer of the present invention is provided wherein a self cleaving dipeptide is linked to the N-terminal alpha amine of the first and / or second incretin peptide wherein the first amino acid of the self-cleaving dipeptide is acylated with a a C16-C30 fatty acid, a C16-C30 phosphonic fatty acid, or a C16-C30 diacid. In one embodiment a incretin peptide / NHR ligand conjugate dimer of the present invention is provided wherein the N-terminal alpha amine of the first and / or second incretin peptide is acylated with a a C16-C30 fatty acid, a C16-C30 phosphonic fatty acid, or a C16-C30 diacid.

[0260] Acylation or alkylation can increase the half-life of the incretin peptide / NHR ligand conjugates in circulation. Acylation or alkylation can advantageously delay the onset of action and / or extend the duration of action at the glucagon, GIP and / or GLP-1 receptors and / or improve resistance to proteases such as DPP-IV and / or improve solubility. Activity at the GLP-1 and / or GIP receptors of the incretin peptide / NHR ligand conjugates may be maintained after acylation. In some embodiments, the potency of the acylated glucagon peptides is comparable to the unacylated versions of the glucagon peptides. In alternative embodiments, the potency of the acylated incretin peptides is increased as compared to that of the unacylated version of the incretin peptides.

[0261] In some embodiments, the first and second incretin peptides of the incretin peptide / NHR ligand conjugates dimer may further comprise a spacer between the acylated amino acid (or at amino acid A of the dipeptide prodrug element) and the acyl group or alkyl group. In some embodiments, the acyl group is a fatty acid or diacid, or salt thereof, e.g. a C4 to C30 fatty acid / diacid, a C4-C30 phosphonic fatty7acid, a C8 to C24 fatty7acid / diacid, a C8-C24 phosphonic fatty acid, cholic acid, a C4 to C30 alkyl, a C8 to C24 alkyl, or an alkyl comprising a steroid moiety of a bile acid. The spacer is any moiety with suitable reactive groups for attaching acyl or alkyl groups. In exemplary embodiments, the spacer comprises an amino acid, a dipeptide, a tripeptide, a hydrophilic bifunctional, or a hydrophobic bifunctional spacer. In some embodiments, the spacer is selected from the group consisting of: Trp, Glu, Asp, Cys and a spacer comprising NH2(CH2CH2O)n(CH2)mCOOH, wherein m is any integer from 1 to 6 and n is any integer from 2 to 12. Such acylated or alkylated glucagon peptides may also further comprise a hydrophilic moiety7, optionally a polyethylene glycol. Any of the foregoing incretin peptide / NHR ligand conjugates may comprise two acyl groups or two alkyl groups linked to the incretin peptides, or a combination thereof.32993-439275

[0262] The acyl group of the incretin peptide / NHR ligand conjugates can be of any size, e.g., any length carbon chain, and can be linear or branched. In some specific embodiments of the invention, the acyl group is a C8 to C30 fatly acid, a C8-C30 phosphonic fatty acid, or C8 to C30 diacid. For example, the acyl group can be any of a C8 fatty acid, CIO fatty acid / diacid, C12 fatty acid / diacid, C14 fatty acid / diacid, C16 fatty acid / diacid, C18 fatty acid / diacid, C20 fatty acid / diacid, C22 fatty acid / diacid, C24 fatty7acid / diacid, C26 fatty7acid / diacid, C28 fatty7acid / diacid, or a C30 fatty7acid / diacid. In some embodiments, the acyl group is a C8 to C20 fatty acid / diacid. e.g., a C18 fatty acid / diacid or a C20 fatty acid / diacid.

[0263] Pharmaceutical compositions comprising the incretin peptide / NHR ligand conjugates, incretin peptide / NHR ligand multimers, or prodrug derivatives thereof, as disclosed herein can be formulated and administered to patients to using standard pharmaceutically acceptable carriers and routes of administration known to those skilled in the art. Accordingly, the present disclosure also encompasses pharmaceutical compositions comprising one or more of the incretin peptide / NHR ligand conjugates, incretin peptide / NHR ligand multimers, or prodrug derivatives thereof, as disclosed herein, in combination with a pharmaceutically acceptable carrier. The pharmaceutical compositions may comprise the incretin peptide / NHR ligand conjugates, incretin peptide / NHR ligand multimers, or prodrug derivatives thereof, as the sole pharmaceutically active component, or the incretin peptide / NHR ligand conjugates, incretin peptide / NHR ligand multimers, or prodrug derivatives thereof, can be combined with one or more additional active agents, including for example an additional anti-obesity peptide. Suitable anti-obesity peptides include those disclosed in US patents 5,691,309. 6,436,435 or US Patent application 20050176643, and including, but not limited to GLP-1, GIP (Gastric Inhibitory Polypeptide), MP1, PYY, MC-4, and Leptin.

[0264] In accordance with some embodiments a pharmaceutical composition is provided comprising any of the novel incretin peptide / NHR ligand conjugates, incretin peptide / NHR ligand multimers, or prodrug derivatives thereof, as disclosed herein, preferably sterile and preferably at a purity level of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, and a pharmaceutically acceptable diluent, carrier or excipient. Such compositions may contain a bioactive peptide prodrug derivative as disclosed herein, wherein the resulting active peptide is present at a concentration of at least 0.5 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 11 mg / ml, 12 mg / ml, 13 mg / ml, 14 mg / ml, 15 mg / ml, 16 mg / ml, 17 mg / ml, 18 mg / ml, 19 mg / ml, 20 mg / ml, 21 mg / ml, 22 mg / ml, 23 mg / ml, 24 mg / ml, 25 mg / ml or higher.

[0265] The pharmaceutical composition can comprise any pharmaceutically acceptable ingredient, including, for example, acidifying agents, additives, adsorbents, aerosol32993-439275propellants, air displacement agents, alkalizing agents, anticaking agents, anticoagulants, antimicrobial preservatives, antioxidants, antiseptics, bases, binders, buffering agents, chelating agents, coating agents, coloring agents, desiccants, detergents, diluents, disinfectants, disintegrants, dispersing agents, dissolution enhancing agents, dyes, emollients, emulsifying agents, emulsion stabilizers, fillers, film forming agents, flavor enhancers, flavoring agents, flow enhancers, gelling agents, granulating agents, humectants, lubricants, mucoadhesives, ointment bases, ointments, oleaginous vehicles, organic bases, pastille bases, pigments, plasticizers, polishing agents, preservatives, sequestering agents, skin penetrants, solubilizing agents, solvents, stabilizing agents, suppository bases, surface active agents, surfactants, suspending agents, sweetening agents, therapeutic agents, thickening agents, tonicity agents, toxicity agents, viscosity -increasing agents, water-absorbing agents, water-miscible cosolvents, water softeners, or wetting agents.L00266] In some embodiments, the pharmaceutical composition comprises any one, or a combination of, the following components: acacia, acesulfame potassium, acetyltributyl citrate, acetyltriethyl citrate, agar, albumin, alcohol, dehydrated alcohol, denatured alcohol, dilute alcohol, aleuritic acid, alginic acid, aliphatic polyesters, alumina, aluminum hydroxide, aluminum stearate, amylopectin, a-amylose, ascorbic acid, ascorbyl palmitate, aspartame, bacteriostatic water for injection, bentonite, bentonite magma, benzalkonium chloride, benzethonium chloride, benzoic acid, benzyl alcohol, benzyd benzoate, bronopol, butylated hydroxyanisole, butylated hydroxytoluene, butylparaben, butylparaben sodium, calcium alginate, calcium ascorbate, calcium carbonate, calcium cyclamate, dibasic anhydrous calcium phosphate, dibasic dehydrate calcium phosphate, tribasic calcium phosphate, calcium propionate, calcium silicate, calcium sorbate, calcium stearate, calcium sulfate, calcium sulfate hemihydrate, canola oil, carbomer, carbon dioxide, carboxymethyl cellulose calcium, carboxymethyl cellulose sodium, p-carotene, carrageenan, castor oil, hydrogenated castor oil, cationic emulsifying wax, cellulose acetate, cellulose acetate phthalate, ethyl cellulose, microcrystalline cellulose, powdered cellulose, silicified microcrystalline cellulose, sodium carboxymethyl cellulose, cetostearyl alcohol, cetrimide, cetyl alcohol, chlorhexidine, chlorobutanol, chlorocresol, cholesterol, chlorhexidine acetate, chlorhexidine gluconate, chlorhexidine hydrochloride, chlorodifluoroethane (HCFC), chlorodifluoromethane, chlorofluorocarbons (CFC)chlorophenoxy ethanol, chloroxylenol, com syrup solids, anhydrous citric acid, citric acid monohydrate, cocoa butter, coloring agents, com oil, cottonseed oil, cresol, m-cresol, o-cresol, p-cresol, croscarmellose sodium, crospovidone, cyclamic acid, cyclodextrins, dextrates. dextrin, dextrose, dextrose anhydrous, diazolidinyl urea, dibutyl phthalate, dibutyl sebacate, diethanolamine, diethyl phthalate, di fluoroethane (HFC), dimethyl-32993-439275P-cyclodextrin, cyclodextrin-type compounds such as Captisol®, dimethyl ether, dimethyl phthalate, dipotassium edentate, disodium edentate, disodium hydrogen phosphate, docusate calcium, docusate potassium, docusate sodium, dodecyl gallate, dodecyltrimethylammonium bromide, edentate calcium disodium, edtic acid, eglumine, ethyl alcohol, ethylcellulose, ethyl gallate, ethyl laurate, ethyl maltol, ethyl oleate, ethylparaben, ethylparaben potassium, ethylparaben sodium, ethyl vanillin, fructose, fructose liquid, fructose milled, fructose pyrogen-free, powdered fructose, fumaric acid, gelatin, glucose, liquid glucose, glyceride mixtures of saturated vegetable fatty acids, glycerin, glyceryl behenate, glyceryl monooleate, glyceryl monostearate, self-emulsifying glyceryl monostearate, glyceryl palmitostearate, glycine, glycols, glycofurol, guar gum, heptafluoropropane (HFC), hexadecyltrimethylammonium bromide, high fructose syrup, human serum albumin, hydrocarbons (HC), dilute hydrochloric acid, hydrogenated vegetable oil, type II, hydroxyethyl cellulose. 2-hydroxyethyl-p-cyclodextrin, hydroxypropyl cellulose, low-substituted hydroxypropyl cellulose, 2-hydroxypropyl-P-cyclodextrin, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, imidurea, indigo carmine, ion exchangers, iron oxides, isopropyl alcohol, isopropyl myristate, isopropyl palmitate, isotonic saline, kaolin, lactic acid, lactitol, lactose, lanolin, lanolin alcohols, anhydrous lanolin, lecithin, magnesium aluminum silicate, magnesium carbonate, normal magnesium carbonate, magnesium carbonate anhydrous, magnesium carbonate hydroxide, magnesium hydroxide, magnesium lauryl sulfate, magnesium oxide, magnesium silicate, magnesium stearate, magnesium trisilicate, magnesium trisilicate anhydrous, malic acid. malt, maltitol, maltitol solution, maltodextrin, maltol, maltose, mannitol, medium chain triglycerides, meglumine, menthol, methylcellulose, methyl methacrylate, methyl oleate, methylparaben, methylparaben potassium, methylparaben sodium, microcrystalline cellulose and carboxymethylcellulose sodium, mineral oil, light mineral oil, mineral oil and lanolin alcohols, oil, olive oil, monoethanolamine, montmorillonite, octyl gallate, oleic acid, palmitic acid, paraffin, peanut oil, petrolatum, petrolatum and lanolin alcohols, pharmaceutical glaze, phenol, liquified phenol, phenoxyethanol, phenoxypropanol, phenylethyl alcohol, phenylmercuric acetate, phenylmercuric borate, phenylmercuric nitrate, polacrilin, polacrilin potassium, poloxamer. polydextrose, polyethylene glycol, polyethylene oxide, polyacrylates, polyethylene-polyoxypropylene-block polymers, polymethacrylates, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitol fatty acid esters, polyoxyethylene stearates, polyvinyl alcohol, polyvinyl pyrrolidone, potassium alginate, potassium benzoate, potassium bicarbonate, potassium bisulfite, potassium chloride, postassium citrate, potassium citrate anhydrous, potassium hydrogen phosphate, potassium metabisulfite, monobasic potassium phosphate, potassium propionate, potassium sorbate, povidone, propanol.32993-439275propionic acid, propylene carbonate, propylene glycol, propylene glycol alginate, propyl gallate, propylparaben, propylparaben potassium, propylparaben sodium, protamine sulfate, rapeseed oil, Ringer's solution, saccharin, saccharin ammonium, saccharin calcium, saccharin sodium, safflower oil, saponite, serum proteins, sesame oil, colloidal silica, colloidal silicon dioxide, sodium alginate, sodium ascorbate, sodium benzoate, sodium bicarbonate, sodium bisulfite, sodium chloride, anhydrous sodium citrate, sodium citrate dehydrate, sodium chloride, sodium cyclamate, sodium edentate, sodium dodecyl sulfate, sodium lauryl sulfate, sodium metabisulfite, sodium phosphate, dibasic, sodium phosphate, monobasic, sodium phosphate, tribasic, anhydrous sodium propionate, sodium propionate, sodium sorbate, sodium starch glycolate, sodium stearyl fumarate, sodium sulfite, sorbic acid, sorbitan esters (sorbitan fatty esters), sorbitol, sorbitol solution 70%, soybean oil, spermaceti wax, starch, com starch, potato starch, pregelatinized starch, sterilizable maize starch, stearic acid, purified stearic acid, stearyl alcohol, sucrose, sugars, compressible sugar, confectioner’s sugar, sugar spheres, invert sugar, Sugartab, Sunset Yellow FCF, synthetic paraffin, talc, tartaric acid, tartrazine, tetrafluoroethane (HFC), theobroma oil, thimerosal, titanium dioxide, alpha tocopherol, tocopheryl acetate, alpha tocopheryl acid succinate, beta-tocopherol, delta-tocopherol, gamma-tocopherol, tragacanth, triacetin, tributyl citrate, triethanolamine, triethyl citrate, trimethyl-0-cyclodextrin. trimethyltetradecylammonium bromide, tris buffer, trisodium edentate, vanillin, type I hydrogenated vegetable oil, water, soft water, hard water, carbon dioxide-free water, pyrogen-free water, water for injection, sterile water for inhalation, sterile water for injection, sterile water for irrigation, waxes, anionic emulsify ing wax, carnauba wax, cationic emulsifying wax, cetyl ester wax, microcrystalline wax, nonionic emulsifying wax, suppository wax, white wax, yellow wax, white petrolatum, wool fat, xanthan gum, xylitol, zein, zinc propionate, zinc salts, zinc stearate, or any excipient in the Handbook of Pharmaceutical Excipients, Third Edition, A. H. Kibbe (Pharmaceutical Press, London, UK, 2000), which is incorporated by reference in its entirety. Remington ’s Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980), which is incorporated by reference in its entirety, discloses various components used in formulating pharmaceutically acceptable compositions and known techniques for the preparation thereof. Except insofar as any conventional agent is incompatible with the pharmaceutical compositions, its use in pharmaceutical compositions is contemplated. Supplementary7active ingredients also can be incorporated into the compositions.

[0267] The pharmaceutical formulations disclosed herein may be designed to be short-acting, fast-releasing, long-acting, or sustained-releasing as described below. The pharmaceutical formulations may also be formulated for immediate release, controlled release32993-439275or for slow release. The instant compositions may further comprise, for example, micelles or liposomes, or some other encapsulated form, or may be administered in an extended release form to provide a prolonged storage and / or delivery effect. The disclosed pharmaceutical formulations may be administered according to any regime including, for example, daily (1 time per day, 2 times per day, 3 times per day, 4 times per day, 5 times per day, 6 times per day), every’ two days, every three days, every four days, every five days, every six days, weekly, bi-weekly, every' three weeks, monthly, or bi-monthly.

[0268] In some embodiments, the foregoing component(s) may be present in the pharmaceutical composition at any concentration, such as, for example, at least A, wherein A is 0.0001% w / v, 0.001% w / v, 0.01% w / v, 0.1% w / v, 1% w / v, 2% w / v, 5% w / v, 10% w / v, 20% w / v, 30% w / v, 40% w / v, 50% w / v, 60% w / v, 70% w / v, 80% w / v, or 90% w / v. In some embodiments, the foregoing component(s) may be present in the pharmaceutical composition at any concentration, such as, for example, at most B, wherein B is 90% w / v, 80% w / v, 70% w / v, 60% w / v, 50% w / v, 40% w / v, 30% w / v, 20% w / v, 10% w / v, 5% w / v, 2% w / v, 1% w / v, 0.1% w / v, 0.001% w / v, or 0.0001%. In other embodiments, the foregoing component(s) may be present in the pharmaceutical composition at any concentration range, such as, for example from about A to about B. In some embodiments, A is 0.0001% and B is 90%.

[0269] The pharmaceutical compositions may be formulated to achieve a physiologically compatible pH. In some embodiments, the pH of the pharmaceutical composition may be at least 5, at least 5.5, at least 6, at least 6.5, at least 7, at least 7.5, at least 8, at least 8.5, at least 9, at least 9.5. at least 10, or at least 10.5 up to and including pH 11. depending on the formulation and route of administration. In certain embodiments, the pharmaceutical compositions may' comprise buffering agents to achieve a physiological compatible pH. The buffering agents may include any compounds capable of buffering at the desired pH such as, for example, phosphate buffers (e.g. PBS), triethanolamine, Tris, bicine. TAPS, tricine, HEPES, TES, MOPS, PIPES, cacodylate, MES, and others. In certain embodiments, the strength of the buffer is at least 0.5 mM, at least 1 mM, at least 5 mM, at least 10 mM, at least 20 mM, at least 30 mM, at least 40 mM, at least 50 mM, at least 60 mM, at least 70 mM, at least 80 mM, at least 90 mM, at least 100 mM, at least 120 mM, at least 150 mM, or at least 200 mM. In some embodiments, the strength of the buffer is no more than 300 mM (e.g. at most 200 mM, at most 100 mM, at most 90 mM, at most 80 mM, at most 70 mM, at most 60 mM, at most 50 mM, at most 40 mM, at most 30 mM, at most 20 mM, at most 10 mM, at most 5 mM, at most 1 mM).

[0270] In some embodiments the pharmaceutical compositions comprise aqueous solutions that are sterilized and optionally stored within various containers. In one embodiment32993-439275aqueous formulations of the incretin peptide / NHR ligand conjugates, multimers, or prodrug derivatives thereof, are prepared at concentrations of about 30mg / ml to about lOOmg / ml in dimethyl sulfoxide (DMSO), including about 30% to 100% DMSO, about 50% to 100% DMSO, about 70% to 100% DMSO, or about 90% to 100% DMSO, with the remainder water. In one embodiment aqueous formulations are prepared in 100% dimethyl sulfoxide. The compounds of the present invention can be used in accordance with some embodiments to prepare pre-formulated solutions ready for injection. In other embodiments the pharmaceutical compositions comprise a lyophilized powder. The pharmaceutical compositions can be further packaged as part of a kit that includes a disposable device for administering the composition to a patient. The containers or kits may be labeled for storage at ambient room temperature or at refrigerated temperature.

[0271] All therapeutic methods, pharmaceutical compositions, kits and other similar embodiments descnbed herein contemplate that the prodrug compounds include all pharmaceutically acceptable salts thereof.

[0272] In some embodiments the kit is provided with a device for administering the incretin peptide / NHR ligand conjugates, multimers, or prodrug derivatives thereof to a patient. The kit may further include a variety of containers, e.g., vials, tubes, bottles, and the like. Preferably, the kits will also include instructions for use. In accordance with some embodiments the device of the kit is an aerosol dispensing device, wherein the composition is prepackaged within the aerosol device. In another embodiment the kit comprises a syringe and a needle, and in some embodiments the prodrug composition is prepackaged within the syringe.

[0273] The present incretin peptide / NHR ligand conjugates, multimers, and prodrug derivatives thereof, have use in the treatment of diabetes, metabolic syndrome and for reducing weight gain or inducing weight loss. Pharmaceutical compositions comprising the incretin peptide / NHR ligand conj ugates, multimers, and prodrug derivatives thereof, and salts thereof, can be administered by any standard route including by oral or inhalation administration.

[0274] Routes of Administration

[0275] The following discussion on routes of administration is merely provided to illustrate exemplary embodiments and should not be construed as limiting the scope in any¬ way.

[0276] Formulations suitable for oral administration can comprise (a) liquid solutions, such as an effective amount of the incretin peptide / NHR ligand conjugates dissolved in diluents, such as water, saline, DMSO, or orange juice; (b) capsules, sachets, tablets, lozenges, and troches, each containing a predetermined amount of the active ingredient, as32993-439275solids or granules; (c) powders; (d) suspensions in an appropriate liquid; and (e) suitable emulsions.

[0277] Liquid formulations may include diluents, such as water, DMSO and alcohols, for example, ethanol, benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant. Capsule forms can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and com starch. Tablet forms can include one or more of lactose, sucrose, mannitol, com starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, colorants, diluents, buffering agents, disintegrating agents, moistening agents, preservatives, flavoring agents, and other pharmacologically compatible excipients. Lozenge forms can comprise the in cretin peptide / NHR ligand conjugates in a flavor, usually sucrose and acacia or tragacanth, as well as pastilles comprising the incretin peptide / NHR ligand conjugates in an inert base, such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, and the like containing, in addition to, such excipients as are known in the art.

[0278] The incretin peptide / NHR ligand conjugates, alone or in combination with other suitable components, can be delivered via pulmonary administration and can be made into aerosol formulations to be administered via inhalation. These aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like. They also may be formulated as pharmaceuticals for non-pressured preparations, such as in a nebulizer or an atomizer. Such spray formulations also may be used to spray mucosa. In some embodiments, the incretin peptide / NHR ligand conjugates are formulated into a powder blend or into microparticles or nanoparticles. Suitable pulmonary7formulations are known in the art. See, e.g., Qian et al., Int J Pharm 366: 218-220 (2009); Adjei and Garren, Pharmaceutical Research, 7(6): 565-569 (1990); Kawashima et al., J Controlled Release 62(1-2): 279-287 (1999); Liu et al., Pharm Res 10(2): 228-232 (1993); International Patent Application Publication Nos. WO 2007 / 133747 and WO 2007 / 141411.

[0279] Formulations suitable for parenteral administration include aqueous and nonaqueous. isotonic sterile injection solutions, which can contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The term, "parenteral" means not through the alimentary canal but by some other route such as subcutaneous, intramuscular, intraspinal, or intravenous. The incretin peptide / NHR ligand conjugates can be administered32993-439275with a physiologically acceptable diluent in a pharmaceutical carrier, such as a sterile liquid or mixture of liquids, including water, saline, aqueous dextrose and related sugar solutions, an alcohol, such as ethanol or hexadecyl alcohol, a glycol, such as propylene glycol or polyethylene glycol, dimethylsulfoxide, glycerol, ketals such as 2,2- dimethyl-153-dioxolane-4-methanol, ethers, poly(ethyleneglycol) 400, oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides with or without the addition of a pharmaceutically acceptable surfactant, such as a soap or a detergent, suspending agent, such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

[0280] Oils, which can be used in parenteral formulations include petroleum, animal, vegetable, or synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, com, olive, petrolatum, and mineral. Suitable fatty acids for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0281] Suitable soaps for use in parenteral formulations include fatty7alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (a) cationic detergents such as. for example, dimethyl dialkyl ammonium halides, and alkyl pyridinium halides, (b) anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylenepolypropylene copolymers, (d) amphoteric detergents such as, for example, alkyl-|3-aminopropionates, and 2-alkyl -imidazoline quaternary ammonium salts, and (e) mixtures thereof.

[0282] The parenteral formulations will typically contain from about 0.5% to about 25% by weight of the incretin peptide / NHR ligand conjugates in solution. Preservatives and buffers may be used. In order to minimize or eliminate irritation at the site of injection, such compositions may contain one or more nonionic surfactants having a hydrophile-hpophile balance (HLB) of from about 12 to about 17. The quantity of surfactant in such formulations will typically range from about 5% to about 15% by weight. Suitable surfactants include polyethylene glycol sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol. The parenteral formulations can be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient, for example, water, for injections, immediately prior to use. Extemporaneous32993-439275injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described.

[0283] Injectable formulations can be used in accordance with the present disclosure. The requirements for effective pharmaceutical carriers for injectable compositions are well-known to those of ordinary skill in the art (see, e.g., Pharmaceutics and Pharmacy Practice, J. B. Lippincott Company, Philadelphia, PA, Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel. 4th ed., pages 622-630 (1986)).

[0284] Additionally, the incretin peptide / NHR ligand conjugates can be made into suppositories for rectal administration by mixing with a variety of bases, such as emulsifying bases or water-soluble bases. Formulations suitable for vaginal administration can be presented as pessaries, tampons, creams, gels, pastes, foams, or spray formulas containing, in addition to the active ingredient, such earners as are known in the art to be appropriate.

[0285] It will be appreciated by one of skill in the art that, in addition to the above- described pharmaceutical compositions, the incretin peptide / NHR ligand conjugates can be formulated as inclusion complexes, such as cyclodextrin inclusion complexes, or liposomes.Heterologous Moiety

[0286] In some embodiments, the incretin peptides of the incretin peptide / NHR ligand conjugates are modified to comprise a heterologous moiety. In some embodiments, the heterologous moiety conjugated to the incretin peptide is a polymer. In some embodiments, the polymer is a biodegradable polymer, including a synthetic biodegradable polymer (e.g., polymers of lactic acid and glycolic acid, polyanhydrides, poly(ortho)esters, polyurethanes, poly(butic acid). poly(valeric acid), and poly(lactide-cocaprolactone)), and a natural biodegradable polymer (e.g., alginate and other polysaccharides including dextran and cellulose, collagen, chemical derivatives thereof (substitutions, additions of chemical groups, for example, alky l, alkylene, hydroxylations, oxidations, and other modifications routinely made by those skilled in the art), albumin and other hydrophilic proteins (e.g., zein and other prolamines and hydrophobic proteins)), as well as any copolymer or mixture thereof. In general, these materials degrade either by enzymatic hydrolysis or exposure to water in vivo, by surface or bulk erosion.

[0287] In some embodiments, the polymer is a water-soluble polymer or a hydrophilic polymer. Hydrophilic polymers are further described herein under “Hydrophilic Heterologous MoietiesP Suitable water-soluble polymers are known in the art and include, for example, polyvinylpyrrolidone, hydroxypropyl cellulose (HPC; Klucel), hydroxypropyl methylcellulose (HPMC; Methocel), nitrocellulose, hydroxypropyl ethylcellulose,32993-439275hydroxypropyl butylcellulose, hydroxypropyl pentylcellulose, methyl cellulose, ethylcellulose (Ethocel), hydroxyethyl cellulose, various alkyd celluloses and hydroxyalkyl celluloses, various cellulose ethers, cellulose acetate, carboxymethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, vinyl acetate / crotonic acid copolymers, poly-hydroxyalkyl methacrylate, hydroxymethyl methacrylate, methacrylic acid copolymers, polymethacrylic acid, polymethylmethacry late, maleic anhydride / methyl vinyl ether copolymers, poly vinyl alcohol, sodium and calcium polyacrylic acid, polyacry lie acid, acidic carboxy polymers, carboxypolymethylene, carboxy vinyl polymers, polyoxyethylene polyoxypropylene copolymer, polymethylvinylether co-maleic anhydride, carboxymethylamide, potassium methacrylate divinylbenzene co-polymer, polyoxyethyleneglycols, polyethylene oxide, and derivatives, salts, and combinations thereof.

[0288] In one embodiment, the polymer is a polyalkylene glycol, including, for example, polyethylene glycol (PEG), for example, of a molecular weight of about 1,000 Daltons to about 40,000 Daltons, or about 20.000 Daltons to about 40,000 Daltons. In one embodiment the PEG polymer is covalently linked to the side chain of an amino acid at position 17, 24 or at the C-terminal amino acid.

[0289] Exemplary embodiments

[0290] In accordance with embodiment 1 a compound is provided comprising the structure Q-L-Y;wherein Q is an incretin peptide;Y is a peroxisome proliferator activated receptor (PPAR) agonist or steroid; and L is a bond or a linking group that links the PPAR agonist or steroid to an amino acid side chain of an amino acid located at position 17, or an amino acid located at a position after position 20, including any amino acid of a C-terminal extension of the incretin peptide, optionally with the proviso that Q is not lanifibranor.

[0291] In accordance with embodiment 2 the compound of embodiment 1 is provided, wherein said linking group is selected from the group consisting of a bond, gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.32993-439275

[0292] In accordance with embodiment 3 the compound of embodiment 1 or 2 is provided wherein L is a bond.

[0293] In accordance with embodiment 4 the compound of any one of embodiments 1-3 is provided, wherein Q is an incretin peptide selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 coagonist, and a glucagon / GIP / GLP-1 tri-agonist, optionally wherein Q is a GIP / GLP-1 coagonist.

[0294] In accordance with embodiment 5 the compound of any one of embodiments 1-4 is provided, wherein Y is linked to Q via the side chain of an amino acid selected from positions 17, 20, 24, 28, 30, 31, 40, the C-terminal amino acid of the incretin peptide and an amino acid added to the C-terminus of Q, optionally wherein Y is linked to Q via the side chain of a C-terminal Lys present at position 40.

[0295] In accordance with embodiment 6 the compound of any one of embodiments 1-5 is provided, wherein Y is a PPAR agonist selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, or Y is a famesoid X receptor (FXR) agonist such as cilofexor.

[0296] In accordance with embodiment 7 the compound of any one of embodiments 1-5 is provided, wherein Y is a steroid selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol.

[0297] In accordance with embodiment 8 the compound of any one of embodiments 1-7 is provided, wherein said incretin peptide comprises the sequence ofi) H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIAWLVX28GRX3i(SEQ ID NO: 28). whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X28 is Arg or Lys; andX31 is Lys, Gly, or a peptide extension comprising the sequence PSSGAPPPSK (SEQ ID NO: 29); orii) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSX4o (SEQ ID NO: 19);111) Y-Aib-QGTFTSDYSIXi3LDKKAQ-Aib-AFlEYLLEGGPSSGAPPPSX4o (SEQ ID NO: 20),wherein X13 is alpha-methyl leucine; or32993-439275iv) H-Aib-QGTFTSDYSKYLDSRRAQDFVQWLM-Z(SEQ ID NO: 33), wherein Z is selected from the group consisting of-COOH, -Asn-COOH, Asn- Thr-COOH. Asn-Thr-W-COOH, and W-COOH. wherein W is 1 to 2 amino acids or GPSSGAPPPSX40 (SEQ ID NO: 34); orv) YX2EGTX6TSDYSIX13LX15X16X17AX19X20AFVQWLIAGGPSSGAPPPSX40-R20 (SEQ ID NO: 43) whereinX2 is an a, a-disubstituted amino acid, optionally Aib;Xe is alpha methylated Phe;X13 is Tyr or an a, a-disubstituted amino acid, optionally Aib;X15 is Glu or Asp;Xi6is Aib, Orn or Lys;X17 is Thr, Vai or He;X19 is Ala or Gin;X20 is an amino acid comprising a (C1-C8 alkyQNEE side chain; andR20 is COOH or CONH2, orvi) X1X2X3GTX6X7SDYSIX13LX15X16X17X18QX20AFVQWLLEGGPSSGAPPPSX40 (SEQ ID NO: 44) whereinXi is Tyr or His;X2 is an a, a-disubstituted amino acid, optionally Aib;X3 is Gin or His;Xe is Phe or alpha methylated Phe;X7 is Thr or (a-Me)Thr;X13 is Tyr, (aMe)Leu, (aMe)Ile, (aMe)Met, (aMe)nLeu, (aMe)Phe or Aib; X15 is Glu or Asp;Xi6is Aib, Orn or Lys;X17 is an amino acid comprising a (C1-C8 alkyl)NH2 side chain;Xi8 is Ala or Tyr;X20 is an a, a-disubstituted amino acid, optionally Aib; andR20 is COOH or CONH2, whereinX40 in any of ii), iii), v) and vi) is independently Lys or is absent.

[0298] In accordance with embodiment 9 the compound of any one of embodiments 1-8 is provided, wherein Y is GW501516, optionally linked to the side chain of an amino acid located at position 24, 29, 31 or 40 of said incretin peptide, or to a C-terminal amide of said incretin peptide.32993-439275

[0299] In accordance with embodiment 10 the compound of any one of embodiments 1-9 is provided, whereinQ comprises the sequence ofi) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSK-R2o (SEQ ID NO: 37); orii) Y-Aib-QGTFTSDYSIXisLDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSK-R20 (SEQ ID NO: 42),wherein X13 is alpha-methyl leucine;R20 is COOH or CONH2. andY is GW501516. linked toi) the side chain of the lysine acid located at position 40 of said incretin peptide, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1. k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4; orii) a C -terminal amide of said incretin peptide, wherein R20 is CONH2, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

[0300] In accordance with embodiment 11, the compound of embodiments 10 is provided, whereini) GW501516 is directly linked to the incretin peptide (L is a bond) or ii) GW501516 is linked to the incretin peptide via a gamma glutamic acid spacer.

[0301] In accordance with embodiment 12 a conjugate is provided comprising an incretin peptide dimer comprising a first and second incretin peptide and one or more NHR ligands covalently linked to the incretin peptide dimer, wherein the first and second incretin peptides are independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 co-agonist, and a Gcg / GIP / GLP-1 tri-agonist to form a heterodimer or homodimer, wherein the incretin peptides are linked together via a bridging linker joining32993-439275i) the side chain of an amino acid independently selected from positions 12, 24, 31 and 40 of the respective first and second incretin peptide, orii) a C-terminal amide of the first incretin peptide to a C-terminal amide of the second incretin peptide; oriii) a C-terminal amide of the first incretin peptide to the side chain of an amino acid selected from positions 12, 24, 31 and 40 of the second incretin peptide,further wherein the one or more nuclear hormone receptor ligands are linked to the incretin heterodimer or homodimer via the side chain of an amino acid at a position not linked to the bridging linker and independently located at position 24, 31 or 40 of one or both of said first and second incretin peptide, optionally at position 40 of one or both of the first and second incretin peptides.

[0302] In accordance with embodiment 13 the conjugate of embodiments 12 is provided, wherein the first and second incretin peptides are independently selected from peptides comprising the sequencei) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSK-R2o (SEQ ID NO: 37); orii) Y-Aib-QGTFTSDYSIXi3LDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSK-R2o (SEQ ID NO: 42, oriii) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPX4o-R2o (SEQ ID NO: 21; oriv) H-Aib-EGTFTSDVSSYLEGQAAKEFIAWLVRGRGX4o-R2o (SEQ ID NO: 22); orv) H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX4o-R2o (SEQ ID NO: 24), whereinR20 is COOH or CONH2;X12 is Ser, Lys or Cys;X13 is alpha-methyl leucine;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys, Om, Cys, or the amino acid at X40 is amidated and linked to -[COCH2(OCH2CH2)k-NH]qwherein k is an integer selected from 1-8 and q is 1 or 2;32993-439275further wherein the first and second incretin peptides are linked together via a bridging linker joining the side chains of amino acids independently selected from position 12, 24, 31 or 40, andsaid nuclear hormone receptor ligand is linked to the side chain of an amino acid at a position not linked to the bridging linker and independently located at position 12, 24 or 40 of said first and / or second incretin peptide, or at a C-terminal primary amine of said first and / or second incretin peptide,optionally wherein the nuclear hormone receptor ligand is linked to a side chain of an amino acid of the first and / or second incretin peptide via a bond or a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

[0303] In accordance with embodiment 14 the conjugate of any one of embodiments 12-13 is provided, wherein said first incretin comprises the amino acid sequenceH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFlX24WLVX28GRPSSGAPPPSX40-R20 (SEQ ID NO: 24, whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys or Cys;R20 is COOH or CONH2; andthe second incretin peptide comprises the amino acid sequenceH-Aib-EGTFTSDVSXi02YLEXi06QAAKEFIX204WLVX208GRPSSGAPPPSX400-R20 (SEQ ID NO: 24), whereinX102 is Ser, Lys or Cys;X106 is Gly or Glu;X204 is Ala. Lys or Cys;X208 is Arg or Lys; andX400 is Lys or Cys;R20 is COOH or CONH2, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 or 24 of said first incretin peptide to the side chain of an amino acid at position 12 or 24 of said32993-439275second incretin peptide, and said nuclear hormone receptor ligand is linked to the side chain of the amino acid at X40 and / or X400, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

[0304] In accordance with embodiment 15 the conjugate of any one of embodiments 12-14 is provided, wherein the bridging linker comprises a structure selected from:i) a disulfide bond;ii) -NH(CO)(CH2)n-S-, wherein n is 1-4: oriii) a triazole.

[0305] In accordance with embodiment 16 the conjugate of any one of embodiments 12-15 is provided, wherein said nuclear hormone receptor ligand is directly linked to the side chain of Lys located at position 40 of said first and / or second incretin peptide.

[0306] In accordance with embodiment 17 the conjugate of any one of embodiments 12-16 is provided, wherein said first incretin peptide comprises the sequence ofH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX2sGRPSSGAPPPSX40-R20 (SEQ ID NO: 24), whereinX12 is Cys;Xi6 is Gly or Glu;X24 is Ala;X28 is Lys; andX40 is Lys or Cys;R20 is COOH or CONH2; andthe second incretin peptide comprises the sequenceH-Aib- EGTFTSDVSX102YLEX106QAAKEFIX204WLVX208GRPSSGAPPPSX400-R20 (SEQ ID NO: 24), whereinX102 is Lys;X106 is Gly or Glu;X204 is Ala;X208 is Lys; andX400 is Lys or Cys;32993-439275R20 is COOH or CONH2, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 of said first incretin peptide to the side chain of an amino acid at position 12 of said second incretin peptide.

[0307] In accordance with embodiment 18 the conjugate of any one of embodiments 12-17 is provided, whereinX40 is absent, Lys or an amino acid comprising an NHR ligand linked to its side chain, optionally via a first spacer; andX400 is an amino acid comprising an NHR ligand linked to its side chain, optionally via a second spacer,wherein said first and second spacer is independently selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

[0308] In accordance with embodiment 19 the conjugate of any one of embodiments 12-18 is provided, wherein X40and X400are each Lys having a NHR ligand linked to the side chain of said Lys, wherein:i) the NHR ligand linked to the Lys of X40 and / or X400 is independently selected from a famesoid X receptor (FXR) agonist, a PPAR agonist or steroid, optionally wherein the PPAR agonist is a compound selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, the FXR agonist is cilofexor and the steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;ii) the NHR ligand linked to Lys of X40 and / or X400 is the same, optionally wherein the NHR ligand is a PPAR agonist selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, a famesoid X receptor (FXR) agonist such as cilofexor, or a steroid selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;iii) the NHR ligand linked to Lys of X40 is a PPAR agonist and the NHR ligand linked to Lys of X400 is a steroid, optionally wherein the PPAR agonist is selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, and the steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;iv) the NHR ligand linked to Lys of X40 is GW501516, and the NHR ligand linked to Lys of X400 is DHEA;32993-439275v) the NHR ligand is linked to both the Lys of X40 and X400, optionally wherein the NHR ligand is GW501516; orvi) the NHR ligand is linked to both the Lys of X40 and X400, optionally wherein the NHR ligand is DHEA.

[0309] In accordance with embodiment 20 the conjugate of any one of embodiments 12-19 is provided, wherein said first incretin comprises the sequence ofH-Aib-EGTFTSDVSCYLEEQAAKEFIAWLVKGRPSSGAPPPSK-R2o (SEQ ID NO: 46); andthe second incretin peptide comprises the sequenceH-Aib-EGTFTSDVSKYLEEQAAKEFIKWLVX28GRPSSGAPPPSK-R2o (SEQ ID NO: 47); whereinR20 is COOH or CONH2, and the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 of said first incretin peptide to the side chain of an amino acid at position 12 of said second incretin peptide via a linker comprising the the structure:-NH(CO)(CH2)2-S-.In accordance with embodiment 21 the conjugate of any one of embodiments 12-20 is provided, wherein the nuclear hormone receptor ligand is linked to the side chain of an amino acid at position 40 of the first and / or second incretin peptide, wherein the nuclear hormone receptor ligand is independently selected from the group consisting of DHEA, estrogen, dexamethasone, GW501516, Pemafibrate, Chiglitazar and Saroglitzazar.

[0310] In accordance with embodiment 22 the conjugate of any one of embodiments 12-21 is provided, wherein said first incretin comprises the sequence ofH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40-R20 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys; andthe second incretin peptide comprises the sequenceH-Aib-EGTFTSDVSXi02YLEXi06QAAKEFIX204WLVX208GRPSSGAPPPSX400-R20 (SEQ ID NO: 24), wherein32993-439275X102 is Ser, Lys or Cys;X106 is Gly or Glu;X204 is Ala. Lys or Cys;X208 is Arg or Lys; andX400 is Lys, andR20 is COOH or CONH2, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chains of the Lys at position 40 of the respective first and second incretin peptides, said bridging linker comprising the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2; and said nuclear hormone receptor ligand is linked to the side chain of an amino acid of the first incretin peptide and / or the second incretin peptide at position independently selected from positions 12, 20, and 24, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

[0311] In accordance with embodiment 23 a conjugate is provided comprising an incretin peptide tetramer comprising two incretin peptide dimers of any one of embodiments 12-22 linked to one another via a tetramer linker, wherein the NHR ligand is linked to one or more of the incretin peptides of the incretin peptide tetramer.

[0312] In accordance with embodiment 24 a conjugate tetramer of embodiment 23 is provided comprising a heterodimer or homodimer of four incretin peptides wherein each of the four incretin peptides are independently selected from SEQ ID NO: 19-24, 37 or 42, wherein the first and second incretin peptides are linked to form a first dimer and the third and fourth incretin peptides are linked to from a second dipeptide, wherein the incretin peptides of the respective first and second dimers are linked via a linker comprising the structure: -[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]s wherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2, and a tetramer linker joins the first and second dimers to form a tetramer wherein the tetramer linker comprises the structure -(CELjmCCHCONEL)-, where m is an integer selected form 1 to 4, that is linked to the primary amine of each of the respective bridging linkers joining the first incretin peptide to the second incretin peptide and the third incretin peptide to the fourth incretin peptide.

[0313] In accordance with embodiment 25 a conjugate tetramer of embodiments 23 or 24 are provided, wherein the first, second, third and fourth incretin peptides are32993-439275independently selected from the sequence H-Aib-EGTFTSDVSX12YLEX16QAAKEFIX24WLVX28GRPSSGAPPPSX40-R20 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys;X40 is Lys; andR20 is COOH or CONH2,wherein the first and second incretin peptides are linked to one another via a first bridging linker joining the side chains of the Lys at position 40 of the respective first and second incretin peptides to form a first dimer, and the third and fourth incretin peptides are linked to one another via a second bridging linker joining the side chains of the Lys at position 40 of the respective third and fourth incretin peptides to form a second dimer,said first and second bridging linkers independently comprising the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2; andan NHR ligand is linked to the side chain of an amino acid of one or more of the first, second, third or fourth incretin peptide at a position independently selected from positions 12, 20, and 24, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acidjz-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4. whereinthe first dimer is linked to the second dimer via a tetramer linker comprises the structure -(Cd l2)m(O ICONI I2)-. where m is an integer selected form 1 to 4, that is linked to the primary amine of each of the respective bridging linkers joining the first incretin peptide to the second incretin peptide and the third incretin peptide to the fourth incretin peptide.In accordance with embodiment 26 a conjugate tetramer is provided wherein 1 to 4, optionally 4, incretin / NHR ligand conjugates independently selected from any one of embodiments 1-11 are covalently linked to a dendritic polymeric core composed of polyethylene glycol, wherein the tetramer has the general structure:32993-439275wherein n is an integer selected from about 100 to about 1000. optionally from about 200 to about 500, further wherein the grey rectangles represent an incretin peptide covalently bound to an NHR ligand.In accordance with embodiment 27, the conjugate tetramer of embodiment 26 is provided wherein each of the covalently linked incretin / NHR ligands of the tetramer are the same.In accordance with embodiment 28, the conjugate tetramer of embodiment 26 is provided wherein two or more of the covalently linked incretin / NHR ligands of the tetramer are different either in the linked incretin peptide, the linked NHR ligand or both.In accordance with embodiment 29, the conjugate tetramer of any one of embodiments 26-28 is provided wherein, the individual incretin peptides, independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 co-agonist, and aglucagon / GIP / GLP-1 tri-agonist, are covalently linked to the dendritic polymeric core via the C-terminus of the incretin peptides.In accordance with embodiment 30, the conjugate tetramer of any one of embodiments 26-29 is provided wherein the NHR ligands are covalently linked, optionally via a spacer, the incretin peptides of the tetramer at positions independently selected from 24, 20, 17, 16 and 12, optionally from positions 24, 20 or 17, optionally from positions 24 and 20, optionally at position 24.In accordance with embodiment 31, the conjugate tetramer of any one of embodiments 26-30 is provided wherein at least one of the incretin peptides is a GIP / GLP-1 co-agonist and at least one of the NHR ligands is GW501516.

[0314] In accordance with embodiment 32 a pharmaceutical composition is provided comprising the conjugate of any one of embodiments 1-11, or a dimer of any one of embodiments 12-22 or a tetramer of any one of embodiments 23-31, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0315] In accordance with embodiment 33 a method of reducing weight gain or inducing weight loss is provided, said method comprising administering to a patient in need32993-439275thereof a pharmaceutical composition of embodiment 32 in an amount effective to reduce weight gain or induce weight loss.

[0316] In accordance with embodiment 34 a method of treating diabetes is provided, said method comprising administering to a patient in need thereof a pharmaceutical composition of embodiment 32 in an amount effective to lower blood glucose levels.

[0317] In accordance with embodiment 35, a prodrug derivative of the incretin peptide / NHR ligand conjugate of any one of embodiments 1-11, or a incretin peptide / NHR ligand conjugate dimer or tetramer of any one of embodiments 12-31 is provided further comprising a dipeptide structure A-B linked to Q of the incretin peptide / NHR ligand conjugate, or the first and / or second incretin peptide of the incretin peptide / NHR ligand conjugate dimer or tetramer, through an amide bond between a carboxyl moiety of B and a primary amine of Q. wherein A-B comprising the structure:wherein(a) Rl, R2, R4 and R8 are independently selected from the group consisting of H, Cl -Cl 8 alkyl, C2-C18 alkenyl, (Cl -Cl 8 alkyljOH, (Cl -Cl 8 alkyljSH, (C2-C3 alkyl)SCH3, (C1-C4 alkyl)CONH2, (C1-C4 alkyljCOOH, (C1-C4 alkyl)NH2, (C1-C4 alkyl)NHC(NH2+)NH2, (C0-C4 alkyl)(C3-C6 cycloalkyl), (C0-C4 alkyl)(C2-C5 heterocyclic), (C0-C4 alkyl)(C6-C10 aryl)R7, (C1-C4 alkyl)(C3-C9 heteroaryl), and C1-C12 alkyl(Wl)Cl-C12 alkyd, wherein W1 is a heteroatom selected from the group consisting of N, S and O;(b) R3 is C1-C8 alky l, or R4 and R3 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;(c) R5 is NH2.

[0318] In accordance with embodiment 36, a prodrug derivative of embodiment 35 is provided whereinRl is (C1-C4 alkyl)NH2,R2, R4 and R8 are H, and A-B is linked to the N-terminal alpha amine of the incretin peptide.

[0319] In accordance with embodiment 37, a prodrug derivative any one of embodiments 35-36 is provided, further comprising a fatty acid or fatty diacid linked to the side32993-439275chain of an amino acid of Q or the side chain of an amino acid of the first and / or second incretin peptide.

[0320] In accordance with embodiment 38, a prodrug derivative of any one of embodiments 35-37 is provided wherein the side chain of the first amino acid of dipeptide A-B is acylated with a fatty acid or fatty diacid, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

Claims

32993-439275WHAT IS CLAIMED:

1. A compound comprising the structure Q-L-Y;wherein Q is an incretin peptide;Y is a NHR ligand, optionally a peroxisome proliferator activated receptor (PPAR) agonist; andL is a bond or a linking group that links the PPAR agonist to an amino acid side chain of an amino acid located at position 17, or an amino acid located at a position after position 20, including any amino acid of a C-terminal extension of the incretin peptide.

2. The compound of claim 1, wherein said linking group is selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

3. The compound of claim 1 wherein L is a bond.

4. The compound of any one of claims 1-3, wherein Q is an incretin peptide selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist. aGIP / GLP-1 co-agonist, and a glucagon / GIP / GLP-1 tri-agonist.

5. The compound of claim 1, wherein Y is linked to Q via the side chain of an amino acid selected from positions 17, 20, 24, 28, 30, 31, 40, the C-terminal amino acid of the incretin peptide and an amino acid added to the C-terminus of Q.

6. The compound of claim 5, wherein said PPAR agonist is selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar.

7. The compound of claim 6 wherein said incretin peptide comprises the sequence of i) H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIAWLVX28GRX31(SEQ ID NO: 28), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X28 is Arg or Lys; and32993-439275X31is Gly, or a peptide extension comprising the sequence PSSGAPPPSK (SEQ ID NO: 29); orii) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSX4o (SEQ ID NO: 19);iii) Y-Aib-QGTFTSDYSIXi3LDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSX4o (SEQ ID NO: 20),wherein X13 is alpha-methyl leucine; oriv) H-Aib-QGTFTSDYSKYLDSRRAQDFVQWLM-Z(SEQ ID NO: 33), wherein Z is selected from the group consisting of-COOH, -Asn-COOH, Asn- Thr-COOH, Asn-Thr-W-COOH, and W-COOH, wherein W is 1 to 2 amino acids or GPSSGAPPPSX40 (SEQ ID NO: 34); orv) YX2EGTX6TSDYSIX13LX15X16X17AX19X20AFVQWLIAGGPSSGAPPPSX40-R20 (SEQ ID NO: 43) whereinX2 is an a,a-disubstituted amino acid, optionally Aib;Xe is alpha methylated Phe;X13 is Tyr or an a,a-disubstituted amino acid, optionally Aib;X15 is Glu or Asp;Xi6is Aib, Orn or Lys;X17 is Thr, Vai or He;X19 is Ala or Gin;X20is an amino acid comprising a (C1-C8 alkyl)NH2side chain; andR20 is COOH or CONH2, orvi) X1X2X3GTX6X7SDYSIX13LX15X16X17X18QX20AFVQWLLEGGPSSGAPPPSX40 (SEQ ID NO: 44) whereinXi is Tyr or His;X2 is an a,a-disubstituted amino acid, optionally Aib;X3 is Gin or His;Xe is Phe or alpha methylated Phe;X7 is Thr or (a-Me)Thr;X13 is Tyr, (aMe)Leu, (aMe)Ile, (aMe)Met, (aMe)nLeu. (aMe)Phe or Aib; X15 is Glu or Asp;Xi6is Aib, Orn or Lys;X17 is an amino acid comprising a (C1-C8 alkyl)NH2 side chain;Xis is Ala or Tyr;X20 is an a,a-disubstituted amino acid, optionally Aib; and32993-439275R20 is COOH or CONH2, whereinX40 in any of ii), iii), v) and vi) is independently Lys or is absent.

8. The compound of any one of claims 1-7 wherein Y is GW501516, optionally linked to the side chain of an amino acid located at position 24, 29, 31 or 40 of said incretin peptide, or to a C-terminal amide of said incretin peptide.

9. The compound of any one of claims 1 -3 whereinQ comprises the sequence of1) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSK (SEQ ID NO: 37); orii) Y-Aib-QGTFTSDYSIXisLDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSK (SEQ ID NO: 42),wherein X13 is alpha-methyl leucine; andY is GW501516, linked toi) the side chain of the lysine acid located at position 40 of said incretin peptide, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4; orii) a C-terminal amide of said incretin peptide, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

10. A conjugate comprising an incretin peptide dimer comprising a first and second incretin peptide linked to one another to form a heterodimer or homodimer, and a NHR ligand covalently linked to said first and / or second incretin peptide, wherein the first and second incretin peptides are independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a GIP receptor agonist, a GIP / GLP-1 co-agonist, and a Gcg / GIP / GLP-1 tri-agonist, wherein the incretin peptides are linked together via a bridging linker joining32993-439275i) the side chain of an amino acid independently selected from positions 12, 24, 31 and 40 of the respective first and second incretin peptide, orii) a C-terminal amide of the first incretin peptide to a C-terminal amide of the second incretin peptide; oriii) a C-terminal amide of the first incretin peptide to the side chain of an amino acid selected from positions 12, 24, 31 and 40 of the second incretin peptide,further wherein said nuclear hormone receptor ligand is linked to said heterodimer or homodimer via the side chain of an amino acid at a position not linked to the bridging linker and independently located at position 24, 31 or 40 of one or both of said first and second incretin peptide.

11. The conjugate of claim 10 wherein the first and second incretin peptides are independently selected from peptides comprising the sequencei) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPSSGAPPPSK (SEQ ID NO: 37); orii) Y-Aib-QGTFTSDYSIXi3LDKKAQ-Aib-AFIEYLLEGGPSSGAPPPSK (SEQ ID NO: 42, oriii) Y-Aib-EGTFTSDYSI-Aib-LDKIAQKAFVQWLIAGGPXro (SEQ ID NO: 21; or iv) H-Aib-EGTFTSDVSSYLEGQAAKEFIAWLVRGRGX4o (SEQ ID NO: 22); or v) H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;X13 is alpha-methyl leucine;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys, Om, Cys, or the amino acid at X40 is amidated and its side chain linked to-[COCH2(OCH2CH2)k-NH]q wherein k is an integer selected from 1-8 and q is 1 or 2; further wherein the first and second incretin peptides are linked together via a bridging linker joining the side chains of amino acids independently selected from position 12, 24, 31 or 40, andsaid nuclear hormone receptor ligand is linked to the side chain of an amino acid at a position not linked to the bridging linker and independently located at position 12, 24 or 40 of32993-439275said first and / or second incretin peptide, or at a C-terminal primary amine of said first and / or second incretin peptide,optionally wherein the nuclear hormone receptor ligand is linked to a side chain of an amino acid of the first and / or second incretin peptide via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

12. The conjugate of claim 11 wherein said first incretin comprises the amino acid sequenceH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24, whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys or Cys; andthe second incretin peptide comprises the amino acid sequenceH-Aib-EGTFTSDVSXi02YLEXi06QAAKEFIX204WLVX208GRPSSGAPPPSX400(SEQ ID NO: 24), whereinX102 is Ser, Lys or Cys;X106 is Gly or Glu;X204 is Ala, Lys or Cys;X208 is Arg or Lys; andX400 is Lys or Cys, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 or 24 of said first incretin peptide to the side chain of an amino acid at position 12 or 24 of said second incretin peptide, and said nuclear hormone receptor ligand is linked to the side chain of the amino acid at X40 and / or X400, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.32993-43927513. The conjugate of claim 11 or 12 wherein the bridging linker comprises a structure selected from:i) a disulfide bond;ii) -NH(CO)(CH2)n-S-, wherein n is 1-4: oriii) a triazole.

14. The conjugate of claim 13 wherein said nuclear hormone receptor ligand is directly linked to the side chain of Lys located at position 40 of said first and / or second incretin peptide.

15. The conjugate of claim 13 or 14 wherein said first incretin peptide comprises the sequence ofH-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40 (SEQ ID NO: 24), whereinX12 is Cys;Xi6 is Gly or Glu;X24 is Ala;X28 is Lys; andX40 is Lys or Cys; andthe second incretin peptide comprises the sequenceH-Aib-EGTFTSDVSXi02YLEXi06QAAKEFIX204WLVX208GRPSSGAPPPSX400 (SEQ ID NO: 24), whereinX102 is Lys;X106 is Gly or Glu;X204 is Ala;X208 is Lys; andX400 is Lys or Cys, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 of said first incretin peptide to the side chain of an amino acid at position 12 of said second incretin peptide.

16. The conjugate of any one of claims 11-15 whereinX40 is absent, Lys or an amino acid comprising an NHR ligand linked to its side chain, optionally via a first spacer; and32993-439275X400is an amino acid comprising an NHR ligand linked to its side chain, optionally via a second spacer,wherein said first and second spacer are each independently selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

17. The conjugate of any one of claims 10-16 wherein X40and X400are each Lys having a NHR ligand linked to the side chain of said Lys, wherein:i) the NHR ligand linked to the Lys of X40 and X400 is independently selected from a PPAR agonist, a famesoid X receptor (FXR) agonist, optionally cilofexor, or steroid, optionally wherein the PPAR agonist is a compound selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, and the steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;ii) the NHR ligand linked to Lys of X40 and X400 is the same, optionally wherein the NHR ligand is a PPAR agonist selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, or a steroid selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol, or the NHR ligand is cilofexor;iii) the NHR ligand linked to Lys of X40 is a PPAR agonist and the NHR ligand linked to Lys of X400 is a steroid, optionally wherein the PPAR agonist is selected from the group consisting of GW501516, Pemafibrate, Chiglitazar and Saroglitzazar, and the steroid is selected from the group consisting of DHEA, testosterone, dexamethasone and estradiol;iv) the NHR ligand linked to Lys of X40 is GW501516, and the NHR ligand linked to Lys of X400 is DHEA;v) the NHR ligand linked to Lys of X40 and X400 is GW501516; orvi) the NHR ligand linked to Lys of X40 and X400 is DHEAvi) the NHR ligand linked to the Lys of of X40 and / or X400 is cilofexor.

18. The conjugate of any one of claims 10-16 wherein said first incretin comprises the sequence ofH-Aib-EGTFTSDVSCYLEEQAAKEFIAWLVKGRPSSGAPPPSK (SEQ ID NO: 46); andthe second incretin peptide comprises the sequence32993-439275H-Aib-EGTFTSDVSKYLEEQAAKEFIKWLVX28GRPSSGAPPPSK (SEQ ID NO: 47); wherein X28is Arg or Lys, optionally X28is Lys, and the first and second incretin peptides are linked to one another via a bridging linker joining the side chain of an amino acid at position 12 of said first incretin peptide to the side chain of an amino acid at position 12 of said second incretin peptide via a linker comprising the the structure:-NH(CO)(CH2)2-S-.

19. The conjugate of any one of claims 10-18 wherein the nuclear hormone receptor ligand is linked to the side chain of an amino acid at position 40 of the first and / or second incretin peptide, wherein the nuclear hormone receptor ligand is independently selected from the group consisting of DHEA, estrogen, dexamethasone, GW501516, Pemafibrate, Chiglitazar and Saroglitzazar.

20. The conjugate of claim 11 wherein said first incretin comprises the sequence of H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX4o (SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys; andthe second incretin peptide comprises the sequenceH-Aib-EGTFTSDVSXi02YLEXi06QAAKEFIX204WLVX208GRPSSGAPPPSX400 (SEQ ID NO: 24), whereinX102 is Ser. Lys or Cys;X106 is Gly or Glu;X204 is Ala, Lys or Cys;X208 is Arg or Lys; andX400 is Lys, wherein the first and second incretin peptides are linked to one another via a bridging linker joining the side chains of the Lys at position 40 of the respective first and second incretin peptides, said bridging linker comprising the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2; and said nuclear hormone receptor ligand is linked to the side chain of an amino acid of the first incretin peptide and / or the second incretin peptide at position independently selected from positions 12,32993-43927520, and 24, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acidjz-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

21. An incretin peptide tetramer comprising two incretin conjugates of any of claims 10-20 linked to one another via a tetramer linker.

22. An incretin peptide / NHR ligand conjugate tetramer comprising four incretin peptides wherein each of the four incretin peptides are independently selected from SEQ ID NO: 16, or 25-30, wherein the first and second incretin peptides are linked to form a first dimer and the third and fourth incretin peptides are linked to from a second dipeptide, wherein the incretin peptides of the respective first and second dimers are linked via a linker comprising the structure: -[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2, and a tetramer linker joins the first and second dimers to form a tetramer wherein the tetramer linker comprises the structure -(CH2)m(CHCONH2)-, where m is an integer selected form 1 to 4, that is linked to the primary amine of each of the respective bridging linkers joining the first incretin peptide to the second incretin peptide and the third incretin peptide to the fourth incretin peptide.

23. An incretin peptide / NHR ligand conjugate tetramer comprising four incretin peptides, wherein the first, second, third and fourth incretin peptides are independently selected from the sequence H-Aib-EGTFTSDVSXi2YLEXi6QAAKEFIX24WLVX28GRPSSGAPPPSX40(SEQ ID NO: 24), whereinX12 is Ser, Lys or Cys;Xi6 is Gly or Glu;X24 is Ala, Lys or Cys;X28 is Arg or Lys; andX40 is Lys;wherein the first and second incretin peptides are linked to one another via a first bridging linker joining the side chains of the Lys at position 40 of the respective first and second incretin peptides to form a first dimer, and the third and fourth incretin peptides are linked to one another via a second bridging linker joining the side chains of the Lys at position 40 of the respective third and fourth incretin peptides to form a second dimer,32993-439275said first and second bridging linkers independently comprising the structure:-[COCH2(OCH2CH2)j-NH]s-(CH2)m(CHCONH2)[NH-(OCH2CH2)j-COCH2]swherein j is an integer selected from 1-8, m is an integer selected from 1-4, and s is 1 or 2; andan NHR ligand is linked to the side chain of an amino acid of one or more of the first, second, third or fourth incretin peptide at a position independently selected from positions 12, 20, and 24, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acid-gamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4, whereinthe first dimer is linked to the second dimer via a tetramer linker comprises the structure -(CH2)m(CHCONH2)-. where m is an integer selected form 1 to 4, that is linked to the primary amine of each of the respective bridging linkers joining the first incretin peptide to the second incretin peptide and the third incretin peptide to the fourth incretin peptide.

24. A conjugate tetramer comprising 4 compounds independently selected from any¬ one of the compounds according to claim 1 covalently linked to a dendritic polymeric core composed of polyethylene glycol, wherein the tetramer has the general structure:wherein n is an integer selected from about 100 to about 1000. optionally from about 200 to about 500, further wherein the grey rectangles each represent an incretin peptide covalently bound to an NHR ligand.

25. The conjugate tetramer of claim 24 wherein each of the covalently linked incretin peptides covalently bound to an NHR ligand are the same.

26. The conjugate tetramer of claim 24 wherein two or more of the covalently linked incretin peptides covalently bound to an NHR ligand are different, either relative to the linked incretin peptide, or relative to the linked NHR ligand, or any combination thereof.32993-43927527. The conjugate tetramer of any one of claims 24-26 wherein the individual incretin peptides are independently selected from the group consisting of a glucagon receptor agonist, a GLP-1 receptor agonist, a G1P receptor agonist, a G1P / GLP-1 co-agonist, and a glucagon / GIP / GLP-1 tri-agonist, wherein the incretin peptides are covalently linked to the dendritic polymeric core via the C-terminus of the incretin peptides.

28. The conjugate tetramer of any one of claims 24-27 is provided wherein the NHR ligands are covalently linked, optionally via a spacer, to the incretin peptides at positions independently selected from 24, 20, 17, 16 and 12, optionally from positions 24, 20 or 17, optionally from positions 24 and 20, optionally at position 24.

29. The conjugate tetramer of any one of claims 24-28 is provided wherein at least one of the incretin peptides is a GIP / GLP-1 co-agonist and at least one of the NHR ligands is GW501 16.

30. A pharmaceutical composition comprising the compound of any one of claims 1-9, or a conjugate of any one of claims 10-20 or a conjugate tetramer of any one of claims 21-29, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

31. A method of reducing weight gain or inducing weight loss, comprising administering to a patient in need thereof a pharmaceutical composition of claim 30 in an amount effective to reduce weight gain or induce weight loss.

32. A method of treating diabetes, comprising administering to a patient in need thereof a pharmaceutical composition of claim 30 in an amount effective to lower blood glucose levels.

33. A prodrug derivative of the compound of any one of claims 1-9, or a conjugate of claims 10-20 or conjugate tetramer of any one of claims 21-29 further comprising a dipeptide structure A-B linked to Q of the incretin peptide / NHR ligand conjugate, or the first and / or second incretin peptide of the incretin peptide / NHR ligand conjugate dimer or tetramer, through an amide bond between a carboxyl moiety of B and a primary amine of Q, wherein A-B comprising the structure:32993-439275wherein(a) R1, R2, R4 and R8 are independently selected from the group consisting of H, Cl -Cl 8 alkyl, C2-C18 alkenyl, (Cl -Cl 8 alkyl)OH, (Cl -Cl 8 alkyl)SH, (C2-C3 alkyl)SCH3, (C1-C4 alkyl)CONH2, (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH2. (C1-C4 alkyl)NHC(NH2+)NH2, (C0-C4 alkyl)(C3-C6 cycloalkyl), (C0-C4 alkyl)(C2-C5 heterocyclic), (C0-C4 alkyl)(C6-C10 aryl)R7, (C1-C4 alkyl)(C3-C9 heteroaryl), and C1-C12 alkyl(W1)C1-C12 alkyl, wherein W1 is a heteroatom selected from the group consisting of N, S and O;(b) R3 is C1-C8 alkyl, or R4 and R3 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;(c) R5 is NH2.

34. The prodrug derivative of claim 33 whereinR1 is (C1-C4 alkyl)NH2,R2, R4 and R8 are H, and A-B is linked to the N-terminal alpha amine of the incretin peptide.

35. The prodrug derivative of any one of claims 33-34, further comprising a fatty acid or fatty diacid linked to the side chain of an amino acid of Q or the side chain of an amino acid of the first and / or second incretin peptide.

36. The prodrug derivative of any one of claims 33-35 wherein the side chain of the first amino acid of dipeptide A-B is acylated with a fatty acid or fatty diacid, optionally via a spacer selected from the group consisting of a gamma glutamic acid, a gamma glutamic acidgamma glutamic acid dipeptide, and a (gamma glutamic acid)z-[COCH2(OCH2CH2)k-NH]q-(gamma glutamic acid)P, wherein z is 0 or 1, k is an integer selected from the range of 2-4 and q and p are independently an integer selected from the range of 0-4.

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