GLP-1 receptor agonists with improved pharmacological and drug delivery properties

Abstract

Disclosed are polypeptides, compositions, and methods useful for activating the glucagon-like peptide-1 (GLP-1) receptor and for treating or preventing a disease or disorder mediated at least in part by glucagon-like peptide 1 (GLP-1).

Description

[Technical field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 210,321, filed June 14, 2021. [Background technology]

[0002] Glucagon-like peptide-1 (GLP-1) is an important brain-gut peptide hormone with well-known physiological functions in glucose metabolism, gastrointestinal secretion and metabolism, and additional therapeutic applications in diabetes, obesity and related metabolic disorders, as well as emerging connections to neurodegenerative diseases (1-16). Historically, GLP-1 was identified as an incretin hormone and shown to enhance meal-induced insulin secretion. GLP-1 is the product of the glucagon gene, which encodes proglucagon. Human GLP-1 is a 30 amino acid peptide derived from preproglucagon, biosynthesized in the gastrointestinal tract (e.g., A-cells in the pancreas and L-cells in the distal ileum) and the brain. Processing of preproglucagon to yield GLP-1(7-36)-NH2 and GLP-2 occurs primarily in L-cells. GLP-1 is normally secreted in response to food intake, and carbohydrates and lipids in particular stimulate GLP-1 secretion.

[0003] GLP-1 has been identified as a very potent and effective stimulator of insulin release. GLP-1 reduces plasma glucagon concentration, slows gastric emptying, stimulates insulin biosynthesis, and enhances insulin sensitivity. In addition, GLP-1 enhances the ability of beta cells to sense and respond to glucose in subjects with impaired glucose tolerance. The insulinotropic effect of GLP-1 in humans increases the rate of glucose metabolism, in part by increasing insulin levels, and in part by enhancing insulin sensitivity. GLP-1 exerts non-insulinotropic actions such as controlling pancreatic beta cell proliferation and survival, bone metabolism, food intake and satiety, enhancing proliferation of neuronal precursor cells and protecting against neuronal apoptosis, reducing cardiac contractility and improving cardiac function after cardiac injury. Taken together, these known pharmacological properties of GLP-1 make it a highly desirable therapeutic agent for the treatment of type II diabetes, obesity and related metabolic disorders and their complications, such as non-alcoholic steatohepatitis, and has a potential role in neurodegenerative diseases and cardiac prevention. Summary of the Invention

[0004] One aspect of the present invention provides polypeptides, compositions, and methods useful for activating the glucagon-like peptide-1 (GLP-1) receptor.

[0005] Thus, the following sequence (I): R XN -X aa 1 -X aa 2 -X aa 3 -X aa 4 -X aa 5 -X aa 6 -X aa 7 -X aa 8 -X aa 9 -X aa 10 -X aa 11 -RYC (I) [In the formula, R XN is H (i.e., des-amino) and -N(Rx)2, where Rx is, independently for each occurrence, H or optionally substituted alkyl, arylalkyl, heteroarylalkyl, formyl, acetyl, alkanoyl, -C(O)-alkyloxy, -C(O)-aryloxy, -C(O)-arylalkyloxy, -C(O)-heterocyclyloxy, -C(O)-heteroarylalkyloxy, -C(O)NH-alkyl, -C(O)NH-aryl, -C(O)NH-aralkyl, -SO2-heterocyclyl, -SO2-alkyl, -SO2-aryl, -SO2-arylalkyl, -SO2-heteroarylalkyl, -SO2-heteroaryl, or ureido; or one occurrence of Rx is hydrogen and the other occurrence is an amino acid residue X aa 0 X is selected from aa 1 is the N-terminal group of; X aa 0 is an optionally substituted amino acid residue selected from Gly, Pro, Arg, Glu, His, Phe, and Trp; X aa 1 is an optionally substituted amino acid residue containing an amino acid side chain containing alkyl, aryl, or heteroaryl; X aa 2 is an optionally substituted amino acid residue selected from Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; X aa 3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 4 is an amino acid residue selected from Gly, Ala, Aib, and β-Ala; Xaa 5 is an optionally substituted amino acid selected from Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; X aa 6 is an optionally substituted amino acid residue disubstituted at the alpha carbon, provided that one of the substituents is an optionally substituted aryl or heteroaryl; X aa 7 is an optionally substituted amino acid residue containing a hydroxyl-containing amino acid side chain; X aa 8 is an optionally substituted amino acid residue selected from Ser, His, and Asn; X aa 9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 10 is an optionally substituted amino acid residue containing a sulfide and / or an optionally substituted aryl or heteroaryl containing amino acid side chain; X aa 11 is an optionally substituted amino acid residue containing a sulfide and / or an optionally substituted aryl or heteroaryl containing amino acid side chain; and R YC is the structure -C(O)N(R Y )2(wherein, R Y is, independently for each occurrence, hydrogen or a PK modifying group; aa 11 is the C-terminal group of Provided herein is a polypeptide represented by:

[0006] Another aspect of the invention relates to a method for treating or preventing a disease or disorder mediated at least in part by glucagon-like peptide 1 in a subject in need of such treatment or prevention, comprising the step of administering to the subject an effective amount of a polypeptide of formula (I).

[0007] A further aspect of the invention relates to a method for treating or preventing diabetes in a subject in need thereof, comprising the step of administering to the subject an effective amount of a polypeptide of sequence (I).

[0008] A further aspect of the invention relates to a method for treating or preventing a neurodegenerative disease in a subject in need thereof, comprising administering to the subject an effective amount of a polypeptide of sequence (I).

[0009] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used to practice or test the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are merely illustrative and are not intended to be limiting.

[0010] Other features, objects, and advantages of the invention will be apparent from the detailed description and claims. [Brief description of the drawings]

[0011] [Figure 1] Correlation between descriptor-based pEC50 calculations and experimental pEC50 results for 28 GLP-1 peptide analogues based on 5nx2-based model structures. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Disclosed herein are polypeptide GLP-1 receptor agonists that exhibit superior pharmacological properties in terms of GLP-1 receptor activation, metabolic stability and pharmacokinetics (by parenteral or oral drug delivery) compared to the native peptide GLP-1. Thus, the disclosed polypeptides are useful for treating or preventing GLP-1-related metabolic disorders, such as type II diabetes and obesity, and neurodegenerative diseases.

[0013] The polypeptides disclosed herein modulate the GLP-1 receptor, for example as agonists or partial agonists of the GLP-1 receptor. The peptides disclosed herein exhibit similar or superior in vivo pharmacological and pharmacokinetic properties compared to GLP-1, making them ideal therapeutic candidates for subcutaneous, oral, pulmonary, nasal, and buccal routes of drug delivery (including sustained release formulations and / or the use of excipients that enhance permeability for uptake into the systemic circulation, depending on the exact route of drug delivery). In particular, the polypeptides disclosed herein, like other agonists of the GLP-1 receptor, exhibit superior postprandial plasma glucose lowering and concomitant increase in plasma insulin levels. Agonists of the GLP-1 receptor have shown clinical benefits in diabetes and its microvascular and macrovascular complications, as well as obesity and related metabolic disorders, and are being evaluated in neurodegenerative diseases, nonalcoholic steatohepatitis (NASH), metabolic disorders in the context of HIV and its treatment, polycystic ovarian syndrome (PCOS), and cardioprotection. Thus, the disclosed polypeptides are effective in treating or preventing complications in type II diabetes and associated metabolic disorders, such as NASH and obesity, as well as neurodegenerative diseases.

[0014] definition For convenience, before further describing the present invention, certain terms used in the specification, examples and appended claims are summarized here. These definitions should be read in light of the remaining parts of this disclosure and understood by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0015] In order that the present invention may be more readily understood, certain terms and phrases are defined below and throughout the specification.

[0016] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0017] The phrase "and / or," as used herein and in the claims, should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctive in some cases and disjunctive in other cases. Multiple elements listed with "and / or" should be construed in the same manner, i.e., "one or more" of the elements so conjoined. Other elements, whether related or unrelated to the elements specifically identified, other than the elements specifically identified by the "and / or" clause may optionally be present. Thus, as a non-limiting example, a reference to "A and / or B," when used with an open-ended expression such as "comprising," may in one embodiment refer to only A (optionally including elements other than B); in another embodiment, it may refer to only B (optionally including elements other than A); in yet another embodiment, it may refer to both A and B (optionally including other elements), etc.

[0018] As used herein and in the claims, "or" should be understood to have the same meaning as "and / or" defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted as being inclusive, i.e., including at least one of, but also including more than one, and optionally including additional items not included in the list. Only terms that are clearly indicated to the contrary, such as "only one of" or "exactly one of" or, when used in the claims, "consisting of", refer to the inclusion of exactly one element of, or of, a list of elements. In general, the term "or", as used herein, shall be interpreted to indicate exclusive alternatives (i.e., "one or the other, but not both") only when preceded by an exclusive term, such as "either", "one of", "only one of", or "exactly one of". "Consisting essentially of", when used in the claims, shall have its ordinary meaning as used in the field of patent law.

[0019] As used herein and in the claims, the phrase "at least one" in reference to a list of one or more elements should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, and not excluding any combination of elements in the list of elements. This definition also allows that elements other than those specifically identified in the list of elements to which the phrase "at least one" refers may optionally be present, whether related or unrelated to the elements specifically identified. Thus, as a non-limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B" or, equivalently, "at least one of A and / or B") can refer, in one embodiment, to at least one A (and optionally including elements other than B), where B is absent and optionally including two or more As; in another embodiment, to at least one B (and optionally including elements other than A), where A is absent and optionally including two or more Bs; in yet another embodiment, to at least one A, and at least one B (and optionally including other elements), optionally including two or more As, and so forth.

[0020] It should also be understood that, unless expressly indicated to the contrary, in any method claimed herein that includes two or more steps or actions, the order of the method steps or actions is not necessarily limited to the order in which the method steps or actions are described.

[0021] In the claims and in the above specification, all transitional phrases, such as "comprising," "including," "holding," "having," "containing," "involving," "holding," "consisting of," and the like, are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedure, Section 2111.03.

[0022] Certain compounds contained in the compositions of the present invention may exist in particular geometric or stereoisomeric forms. In addition, the compounds of the present invention may also be optically active. The present invention contemplates all such compounds as being within the scope of the present invention, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof. Additional asymmetric carbon atoms may be present in a substituent, such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in the present invention.

[0023] "Geometric isomer" means an isomer that differs in the orientation of substituent atoms in relation to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. The atoms (other than H) at each end of a carbon-carbon double bond may be in the E (substituents are on opposite sides of the carbon-carbon double bond) or Z (substituents are oriented on the same side) configuration. "R", "S", "S*", "R*", "E", "Z", "cis", and "trans" indicate the configuration relative to the core molecule. Some of the disclosed compounds may exist in "atropisomeric" forms or as "atropisomers". Atropisomers are stereoisomers that result from hindrance of rotation about a single bond, where the steric strain hindrance to rotation is high enough to allow for the isolation of conformers. The compounds of the present invention may be prepared as individual isomers by synthesis specific for either isomer, or resolved from a mixture of isomers. Classical resolution techniques include forming a salt of the free base of each isomer of the isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming a salt of the acid form of each isomer of the isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of the isomeric pair using optically pure acids, amines or alcohols (followed by chromatographic separation and removal of the chiral auxiliary), or resolving the isomeric mixture of starting materials or final products using a variety of well-known chromatographic methods.

[0024] For example, if a particular enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or by derivatization with an asymmetric auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, if the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, a diastereomeric salt may be formed with a suitable optically active acid or base, and the diastereomers thus formed may then be resolved by fractional crystallization or chromatographic means well known in the art, followed by recovery of the pure enantiomers.

[0025] Percent purity by mole fraction is the ratio of moles of an enantiomer (or diastereomer) to moles of the enantiomer (or diastereomer) and its optical isomer. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least about 60%, about 70%, about 80%, about 90%, about 99%, or about 99.9% pure by mole fraction compared to the other stereoisomer. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least about 60%, about 70%, about 80%, about 90%, about 99%, or about 99.9% pure by mole fraction. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least about 60%, about 70%, about 80%, about 90%, about 99%, or about 99.9% pure by mole fraction.

[0026] When a disclosed compound is named or depicted by structure without indicating stereochemistry, and the compound has at least one asymmetric center, it should be understood that the name or structure encompasses any enantiomer of the compound without the corresponding optical isomer, a racemic mixture of the compound, or a mixture enriched in one enantiomer compared to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating stereochemistry, and the compound has two or more asymmetric centers, it should be understood that the name or structure encompasses a diastereomer without the other diastereomer, some diastereomers without other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers enriched in one diastereomer compared to the other diastereomer(s), or mixtures of diastereomers enriched in one or more diastereomers compared to the other diastereomers. The present invention encompasses all of these forms.

[0027] Structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, hydrogen is replaced with deuterium or tritium, or carbon is replaced with13 C- or 14 Compounds produced by substituting C-enriched carbons are within the scope of the present invention.

[0028] The term "prodrug" as used herein includes compounds that are converted to therapeutically active agents under physiological conditions. A common method for making a prodrug is to include selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by enzymatic activity of the host animal.

[0029] The phrase "pharmacologically acceptable excipient" or "pharmacologically acceptable carrier" as used herein means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, that is involved in the carrying or transport of a subject chemical entity from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation, not injurious to the patient, and substantially non-pyrogenic. Some examples of materials which may function as pharma- ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and the like. Soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical formulations. In certain embodiments, the pharmaceutical compositions of the present invention are non-pyrogenic, i.e., do not induce a significant increase in body temperature when administered to a patient.

[0030] The term "pharmaceutically acceptable salts" refers to the relatively non-toxic, inorganic and organic acid addition salts of a compound(s). These salts can be prepared in situ during the final isolation and purification of the compound(s), or can be prepared by separately reacting the purified compound(s) in free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulfonate salts (see, e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0031] In other cases, the compounds useful in the methods of the present invention may contain one or more acidic functional groups and therefore can form pharma-ceutically acceptable salts with pharma-ceutically acceptable bases. The term "pharma-ceutically acceptable salts" in these instances refers to the relatively non-toxic inorganic and organic base addition salts of the compound(s). These salts can likewise be prepared in situ during the final isolation and purification of the compound(s), or can be prepared by separately reacting the purified compound(s) in free acid form with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharma-ceutically acceptable metal cation, with ammonia, or with a pharma-ceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra).

[0032] The term "pharmaceutically acceptable cocrystal" refers to a solid-state conformer that does not form formal ionic interactions with a small molecule.

[0033] A "therapeutically effective amount" (or "effective amount") of a compound for use in therapy refers to the amount of compound in a formulation that, when administered (to a mammal, preferably a human) as part of a desired dosing regimen, alleviates the symptoms, improves the condition, or delays the onset of a disease state, e.g., in accordance with clinically accepted standards or cosmetic purposes for the disorder or condition being treated, at a reasonable benefit / risk ratio applicable to any medical treatment.

[0034] The term "prophylactic or therapeutic" treatment is art-recognized and includes administration to a host of one or more of the subject compositions. When a subject composition is administered prior to the clinical manifestation of an undesirable condition (e.g., a disease or other undesirable condition in a host animal), the treatment is prophylactic (i.e., it protects the host against the onset of the undesirable condition), whereas when the composition is administered after the manifestation of an undesirable condition, the treatment is therapeutic (i.e., it is intended to reduce, ameliorate, or stabilize an existing undesirable condition or its side effects).

[0035] The term "patient" or "subject" refers to a mammal in need of a particular treatment. In certain embodiments, the patient is a primate, dog, cat, or horse. In certain embodiments, the patient is a human.

[0036] Aliphatic chains include the classes alkyl, alkenyl, and alkynyl, as defined below. Straight aliphatic chains are limited to unbranched carbon chain moieties. As used herein, the term "aliphatic group" refers to straight, branched, or cyclic aliphatic hydrocarbon groups, including saturated and unsaturated aliphatic groups, such as alkyl, alkenyl, or alkynyl groups.

[0037] "Alkyl" refers to a fully saturated, cyclic or acyclic, branched or unbranched carbon chain moiety having the specified number of carbon atoms, or up to 30 carbon atoms if not specified. For example, alkyl of 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, as well as moieties that are positional isomers of these moieties. Alkyl of 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, and tetracosyl. In certain embodiments, a straight or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C for straight chain). 30 , for branched chains C3~C 30 ), more preferably having 20 or fewer carbon atoms. Alkyl groups can be substituted or unsubstituted.

[0038] As used herein, the term "heteroalkyl" refers to an alkyl moiety, as defined herein above, that contains one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of a carbon atom.

[0039] As used herein, the term "haloalkyl" refers to an alkyl group, as defined herein above, that is substituted with at least one halogen.

[0040] As used herein, the term "hydroxyalkyl" refers to an alkyl group, as defined herein above, substituted with at least one hydroxyl.

[0041] As used herein, the term "alkylene" refers to an alkyl group having a specified number of carbons, e.g., 2-12 carbon atoms, containing two points of attachment to the remainder of the compound on its longest carbon chain. Non-limiting examples of alkylene groups include methylene-(CH2)-, ethylene-(CH2CH2)-, n-propylene-(CH2CH2CH2)-, isopropylene-(CH2CH(CH3))-, and the like. Alkylene groups may be cyclic or acyclic, branched or unbranched carbon chain moieties, and may be optionally substituted with one or more substituents.

[0042] "Cycloalkyl" means mono- or bi-cyclic or bridged or spiro- or polycyclic saturated carbocyclic rings, each of which has from 3-12 carbon atoms. Preferred cycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 3-6 carbons in the ring structure. Cycloalkyl groups can be substituted or unsubstituted.

[0043] As used herein, the term "halocycloalkyl" refers to a cycloalkyl group, as defined herein above, that is substituted with at least one halogen.

[0044] "Cycloheteroalkyl" refers to a cycloalkyl moiety, as defined herein above, that contains one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of a carbon atom. Preferred cycloheteroalkyls have from 4-8 carbon atoms and heteroatoms in their ring structure, and more preferably have 4-6 carbon and heteroatoms in the ring structure. Cycloheteroalkyl groups can be substituted or unsubstituted.

[0045] "Ureido" refers to an optionally substituted urea moiety, e.g., -NHC(O)NH2 or -NHC(O)NHR, where R is alkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl.

[0046] "PK modifier group" refers to a group that modifies, e.g., improves, the pharmacokinetic (PK) profile of the polypeptide to which it is attached. The PK modifier group may utilize a known binding agent for human serum albumin (HSA). This interaction with albumin may result in reduced in vivo clearance. Thus, an example of a PK modifier group is a serum albumin binding group.

[0047] Unless the number of carbons is otherwise specified, "lower alkyl," as used herein, means an alkyl group as defined above, but having from 1 to 10 carbons, more preferably from 1 to 6 carbon atoms in its backbone structure, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Similarly, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Throughout this application, preferred alkyl groups are lower alkyls. In certain embodiments, a substituent designated herein as alkyl is a lower alkyl.

[0048] "Alkenyl" refers to any cyclic or acyclic, branched or unbranched, unsaturated carbon chain moiety having the specified number of carbon atoms, or up to 26 carbon atoms if no limit to the number of carbon atoms is specified, and having one or more double bonds in the moiety. Alkenyls of 6 to 26 carbon atoms are exemplified by hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosenyl, docosenyl, tricosenyl, and tetracosenyl in their various isomeric forms, where the unsaturated bond(s) may be located anywhere in the moiety and may have either the (Z) or (E) configuration around the double bond(s).

[0049] "Alkynyl" refers to a hydrocarbyl moiety within the scope of alkenyl, but which has one or more triple bonds.

[0050] The term "aryl" as used herein includes 3-12 membered substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon (i.e., carbocyclic aryl) or one or more atoms are heteroatoms (i.e., heteroaryl). Preferably, the aryl group includes a 5-12 membered ring, more preferably a 6-10 membered ring. The term "aryl" also includes polycyclic ring systems having two or more rings in which two or more carbons are common to two adjacent rings, at least one of the rings being aromatic, for example, the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Carbocyclic aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like. Heteroaryl groups include substituted or unsubstituted aromatic 3-12 membered ring structures, more preferably 5-12 membered rings, more preferably 5-10 membered rings, and the ring structures include 1-4 heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, etc. Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic.

[0051] The terms "halo", "halide", or "halogen" as used herein means halogen, including, for example, but not limited to, fluoro, chloro, bromo, iodo, and the like, in both radioactive and non-radioactive forms. In preferred embodiments, halo is selected from the group consisting of fluoro, chloro, and bromo.

[0052] The terms "heterocyclyl" or "heterocyclic group" refer to a 3- to 12-membered ring structure, more preferably a 5- to 12-membered ring, and more preferably a 5- to 10-membered ring, whose ring structure includes one to four heteroatoms. The heterocycle can be monocyclic, bicyclic, spirocyclic, or polycyclic. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like. The heterocyclic ring can be substituted at one or more positions with substituents as described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, -CF3, -CN, and the like.

[0053] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It is understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is in accordance with the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound that does not spontaneously undergo transformation, for example, by rearrangement, cyclization, elimination, and the like. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents may be one or more and may be the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. Substituents include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. In preferred embodiments, the substituent on the substituted alkyl is C 1~6 Alkyl, C 3~6 In a more preferred embodiment, the substituent on the substituted alkyl is selected from fluoro, carbonyl, cyano, or hydroxyl. It will be understood by those skilled in the art that the substituents themselves can be substituted, where appropriate. Unless specifically stated as "unsubstituted," references to chemical moieties herein are understood to include substituted variants. For example, references to "aryl" groups or moieties implicitly include both substituted and unsubstituted variants.

[0054] As used herein, the definition of each expression, e.g., alkyl, m, n, etc., when it occurs more than one time in any structure, is intended to be independent of its definition elsewhere in the same structure.

[0055] As used herein, "small molecule" refers to a small organic or inorganic molecule with a molecular weight below about 3,000 Daltons. Generally, small molecules useful in the present invention have a molecular weight less than 3,000 Daltons (Da). A small molecule can be, for example, at least about 100 Da to about 3,000 Da (e.g., between about 100 Da and about 3,000 Da, between about 100 Da and about 2500 Da, between about 100 Da and about 2,000 Da, between about 100 Da and about 1,750 Da, between about 100 Da and about 1,500 Da, between about 100 Da and about 1,250 Da, between about 100 Da and about 1,000 Da, between about 100 Da and about 750 Da, between about 100 Da and about 500 Da, between about 200 and about 1500, between about 500 and about 1000, between about 300 Da and about 1000 Da, or between about 100 Da and about 250 Da).

[0056] In some embodiments, "small molecule" refers to an organic, inorganic, or organometallic compound, typically having a molecular weight of less than about 1000. In some embodiments, small molecules are organic compounds having a size on the order of 1 nm. In some embodiments, small molecule drugs of the present invention include oligopeptides and other biomolecules having a molecular weight of less than about 1000.

[0057] An "effective amount" is an amount sufficient to produce a beneficial or desired result. For example, a therapeutic amount is an amount that achieves a desired therapeutic effect. This amount may be the same as or different from a prophylactically effective amount, which is the amount necessary to prevent the onset of a disease or disease symptoms. An effective amount can be administered in one or more administrations, applications, or dosages. A therapeutically effective amount of a composition will depend on the composition selected. The composition can be administered from once or more times a day to once or more times a week; for example, every other day. Those skilled in the art will understand that certain factors, including but not limited to the severity of the disease or disorder, previous treatments, the general health and / or age of the subject, and other diseases present, can affect the dosage and timing required to effectively treat a subject. Furthermore, treatment of a subject with a therapeutically effective amount of a composition described herein can include a single treatment or a series of treatments.

[0058] The terms "reduce," "reduce," "reduced," "reduction," "decrease," and "inhibit" are all used generally herein to mean a statistically significant amount of reduction compared to a reference. However, for the avoidance of doubt, "reduce", "reduction" or "decrease" or "inhibit" typically means a decrease of at least 10% compared to a reference level, and may include, for example, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99% decrease in a given entity or parameter compared to a reference level, for example up to (including complete absence), or any decrease between 10-99% compared to the absence of a given treatment.

[0059] The terms "increased", "increase" or "enhance" or "activate" are all used herein to generally mean an increase by a statistically significant amount; for the avoidance of any doubt, the terms "increased", "increase" or "enhance" or "activate" mean an increase of at least 10% compared to a reference level, such as at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% increase, or up to and including a 100% increase, or any increase between 10-100% compared to a reference level, or at least about 2-fold, or at least about 3-fold, or at least about 4-fold, or at least about 5-fold, or at least about 10-fold increase compared to a reference level, or any increase between or greater than 2-fold.

[0060] As used herein, the term "modulate" includes upregulation and downregulation, e.g., enhancing or inhibiting a response.

[0061] "Radiopharmaceuticals" as defined herein refer to pharmaceuticals that contain at least one radiation-emitting radioisotope. Radiopharmaceuticals are routinely used in nuclear medicine for the diagnosis and / or therapy of various diseases. Radiolabeled pharmaceuticals, such as radiolabeled antibodies, contain a radioisotope (RI) that serves as a radiation source. As contemplated herein, the term "radioisotope" includes metallic and non-metallic radioisotopes. The radioisotope is selected based on the medical use of the radiolabeled pharmaceutical. When the radioisotope is a metallic radioisotope, typically a chelating agent is used to bind the metallic radioisotope to the remainder of the molecule. When the radioisotope is a non-metallic radioisotope, the non-metallic radioisotope is typically linked to the remainder of the molecule directly or via a linker.

[0062] The term "diabetes and related diseases or conditions" refers to, but is not limited to, type II diabetes, type I diabetes, impaired glucose tolerance, obesity, hyperglycemia, Syndrome X, metabolic syndrome, diabetic complications, and hyperinsulinemia.

[0063] For purposes of this invention, the chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.

[0064] Compounds of the Invention One aspect of the invention is the following sequence (I): R XN -X aa 1 -X aa 2 -X aa 3 -X aa 4 -X aa 5 -X aa 6 -X aa 7 -X aa 8 -X aa 9 -X aa 10 -X aa 11 -R YC (I) [In the formula, R XNis H (i.e., des-amino) and -N(Rx)2, where Rx is, independently for each occurrence, H or optionally substituted alkyl, arylalkyl, heteroarylalkyl, formyl, acetyl, alkanoyl, -C(O)-alkyloxy, -C(O)-aryloxy, -C(O)-aralkyloxy, -C(O)-heterocyclyloxy, -C(O)-heteroarylalkyloxy, -C(O)NH-alkyl, -C(O)NH-aryl, -C(O)NH-arylalkyl, -SO2-heterocyclyl, -SO2-alkyl, -SO2-aryl, -SO2-arylalkyl, -SO2-heteroarylalkyl, -SO2-heteroaryl, or ureido; or one occurrence of Rx is hydrogen and the other occurrence is an amino acid residue X aa 0 Xaa is selected from 1 is the N-terminal group of; X aa 0 is an optionally substituted amino acid residue selected from Gly, Pro, Arg, Glu, His, Phe, and Trp; X aa 1 is an optionally substituted amino acid residue containing an amino acid side chain containing alkyl, aryl, or heteroaryl; X aa 2 is an optionally substituted amino acid residue selected from Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; X aa 3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 4 is an amino acid residue selected from Gly, Ala, Aib, and β-Ala; X aa 5is an optionally substituted amino acid selected from Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; X aa 6 is an optionally substituted amino acid residue disubstituted at the alpha carbon, provided that one of the substituents is an optionally substituted aryl or heteroaryl; X aa 7 is an optionally substituted amino acid residue containing a hydroxyl-containing amino acid side chain; X aa 8 is an optionally substituted amino acid residue selected from Ser, His, and Asn; X aa 9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 10 is an optionally substituted amino acid residue containing a sulfide and / or an optionally substituted aryl or heteroaryl containing amino acid side chain; X aa 11 is an optionally substituted amino acid residue containing a sulfide and / or an optionally substituted aryl or heteroaryl containing amino acid side chain; and R YC is the structure -C(O)N(R Y )2(wherein, R Y is, independently for each occurrence, hydrogen or a PK modifying group; aa 11 is the C-terminal group of The present invention relates to a polypeptide represented by

[0065] In certain embodiments, R YC is the structure -C(O)N(R Y )2[where, R Y is, independently for each occurrence, hydrogen or a serum albumin binding group; aa11 is the C-terminal group of

[0066] In certain embodiments, X aa 1 is an optionally substituted amino acid residue containing an amino acid side chain containing (C1-C4) alkyl, imidazole, or phenyl.

[0067] In certain embodiments, X aa 1 is an optionally substituted amino acid residue selected from Leu, His, and Tyr.

[0068] In certain embodiments, the amino acid residues, if substituted, are substituted with at least one halo, hydroxyl, or alkyl.

[0069] In certain embodiments, X aa 2 is an unsubstituted amino acid residue selected from Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, and D-Val.

[0070] In certain embodiments, X aa 2 is a substituted amino acid residue selected from Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, and D-Val.

[0071] In certain embodiments, the amino acid residue is selected from Aib, Pro, α-methyl-Pro, and Val.

[0072] In certain embodiments, the amino acid residue is substituted with at least one halo or alkyl.

[0073] In certain embodiments, X aa 3 is an optionally substituted amino acid residue containing a carboxyl-containing amino acid side chain.

[0074] In certain embodiments, X aa 3 is an amino acid residue selected from Asp and Glu.

[0075] In certain embodiments, X aa 3 is an optionally substituted amino acid residue comprising an amino acid side chain containing a sulfonic acid group.

[0076] In certain embodiments, X aa 3 is an amino acid residue selected from cysteic acid.

[0077] In certain embodiments, the amino acid residues, if substituted, are substituted with at least one halo or alkyl.

[0078] In certain embodiments, X aa 4 is an amino acid residue selected from Gly and Ala.

[0079] In certain embodiments, X aa 5 is an unsubstituted amino acid residue selected from Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal.

[0080] In certain embodiments, X aa 5 is a substituted amino acid residue selected from Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal.

[0081] In certain embodiments, X aa 5 is unsubstituted or substituted Thr.

[0082] In certain embodiments, X aa 5is substituted with at least one halo or alkyl.

[0083] In certain embodiments, X aa 6 teeth, [ka] [In the formula, X 6a is alkyl; and X 6b is a substituted arylalkyl. The amino acid residue is an optionally substituted amino acid residue represented by the following formula:

[0084] In certain embodiments, the arylalkyl is substituted with at least one halo.

[0085] In certain embodiments, X 6a is methyl; and X 6b is benzyl, 2-fluorobenzyl, or 2,4-difluorobenzyl.

[0086] In certain embodiments, X aa 6 is an optionally substituted amino acid residue selected from α-MePhe, α-MePhe(2-F), and α-MePhe(2,6-DiF).

[0087] In certain embodiments, X aa 7 is an optionally substituted amino acid residue selected from Thr, α-MeThr, Ser, and α-MeSer.

[0088] In certain embodiments, the amino acid residues, if substituted, are substituted with at least one halo or alkyl.

[0089] In certain embodiments, X aa 8 is an unsubstituted amino acid residue selected from Ser, His, and Asn.

[0090] In certain embodiments, X aa 8 is a substituted amino acid residue selected from Ser, His, and Asn.

[0091] In certain embodiments, X aa 8 is unsubstituted or substituted Ser.

[0092] In certain embodiments, X aa 8 When substituted, it is substituted with at least one halo or alkyl.

[0093] In certain embodiments, X aa 9 is an optionally substituted amino acid residue containing a carboxyl-containing amino acid side chain.

[0094] In certain embodiments, X aa 9 is an amino acid residue selected from Asp and Glu.

[0095] In certain embodiments, X aa 9 is an optionally substituted amino acid residue comprising an amino acid side chain containing a sulfonic acid group.

[0096] In certain embodiments, X aa 9 is an optionally substituted cysteic acid.

[0097] In certain embodiments, the amino acid residues, if substituted, are substituted with at least one halo or alkyl.

[0098] In certain embodiments, X aa 10 is an amino acid residue that includes an amino acid side chain that contains a substituted aryl.

[0099] In certain embodiments, X aa10 is further substituted at the α-carbon. In certain embodiments, X aa 10 is further substituted at the α-carbon with alkyl. aa 10 is further substituted at the α-carbon with methyl.

[0100] In certain embodiments, X aa 10 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X3 is N or CR 10d and; X4 is N or CR 10e and; X5 is N or CR 10f and; Z1 is absent or present, and, if present, is S or SO2; R 10a is H or alkyl; and R 10b , R 10c , R 10d , R 10e , and R 10f are independently selected from H, halogen, and alkyl. It is expressed by:

[0101] In certain embodiments, X aa 10 teeth, [ka] It is expressed by:

[0102] In certain embodiments, X aa 10 teeth, [ka] It is expressed by:

[0103] In certain embodiments, X aa 10 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; X6 is N or CR 10g and; X7 is N or CR 10h and; X8 is N or CR 10i and; X9 is N or CR 10j and; X 10 is N or R 10k and; Z1 is absent or present, and, if present, is S or SO2; Z2 is absent or present, and, if present, is S or SO2; R 10a is selected from H and alkyl; and R 10b , R 10c , R 10e , R 10f , R 10g , R 10h , R 10i , R 10j , and R 10k are independently selected from H, halogen, and alkyl. It is expressed by:

[0104] In certain embodiments, X aa 10 teeth, [ka] It is expressed by:

[0105] In certain embodiments, X aa 10 teeth, [ka] Represented by TIFF2024525236000009.tif54144.

[0106] In certain embodiments, X aa 10 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; Z1 is absent or present and, if present, is selected from CH2, S, O, NH, SO2, SO2-NH, NH-SO2, NHC(O), and C(O)NH; R 10a is selected from H and alkyl; R 10d is -OH or -L1-L2-L3-R 10d 'And; R 10d ' is -NH2, (C1-C 20 (alkyl)-COH or optionally substituted (C1-C6 alkyl)-aryl; L1 is absent or present, and if present, is a linker; L2 is a linker comprising an ether moiety; L3 is absent or present, and if present, is a linker comprising an amino acid moiety; and R 10b , R10c , R 10d , R 10e , and R 10f are independently selected from H, halogen, and alkyl. It is expressed by:

[0107] In certain embodiments, R 10d -L1-L2-L3-R 10d ' is.

[0108] In certain embodiments, X aa 10 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; X6 is N or CR 10g and; X7 is N or CR 10h and; X9 is N or CR 10j and; X 10 is N or CR 10k and; Z1 is absent or present and, if present, is selected from CH2, NH, S, SO2, O, SO2-NH, NH-SO2, NHC(O), and C(O)NH; Z2 is absent or present and, if present, is selected from NH, S, SO2, O, SO2-NH, NH-SO2, NHC(O), and C(O)NH; R 10a is selected from H and alkyl; R 10i -L1-L2-L3-R 10i 'And; R 10i ' is -NH2, (C1-C 20(alkyl)-COH or optionally substituted (C1-C6 alkyl)-aryl; L1 is absent or present, and if present, is a linker; L2 is a linker comprising an ether moiety; L3 is absent or present, and if present, is a linker comprising an amino acid moiety; and R 10b , R 10c , R 10e , R 10f , R 10g , R 10h , R 10j , and R 10k are independently selected from H, halogen, and alkyl. It is expressed by:

[0109] In certain embodiments, X aa 10 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; R 10a is selected from H and alkyl; R 10b , R 10c , R 10e , and R 10f is independently selected from H, halogen, and alkyl; and Z3 is a substituted heteroaryl. It is expressed by:

[0110] In certain embodiments, Z3 is a 5-membered substituted heteroaryl.

[0111] In certain embodiments, the 5-membered substituted heteroaryl is a substituted triazolyl.

[0112] In certain embodiments, the substituted heteroaryl is -(C 1~20 It is substituted with -COH.

[0113] In certain embodiments, the substituted heteroaryl is -(C 10~20 It is substituted with -COH.

[0114] In certain embodiments, the substituted heteroaryl is -(C 15 It is substituted with -COH.

[0115] In certain embodiments, X aa 10 teeth, [ka] It is expressed by:

[0116] In certain embodiments, X aa 11 is an amino acid residue that includes an amino acid side chain that contains a substituted aryl.

[0117] In certain embodiments, X aa 11 is further substituted at the α-carbon. In certain embodiments, X aa 11 is further substituted at the α-carbon with alkyl. aa 11 is further substituted at the α-carbon with methyl.

[0118] In certain embodiments, X aa 11 teeth, [ka] [In the formula, X1 is N or CR10b and; X2 is N or CR 10c and; X3 is N or CR 10d and; X4 is N or CR 10e and; X5 is N or CR 10f and; Z1 is absent or present, and, if present, is S or SO2; R 10a is selected from H and alkyl; and R 10b , R 10c , R 10d , R 10e , and R 10f are independently selected from H, halogen, and alkyl. It is expressed by:

[0119] In certain embodiments, X aa 11 teeth, [ka] It is expressed by:

[0120] In certain embodiments, X aa 11 teeth, [ka] It is expressed by:

[0121] In certain embodiments, X aa 11 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; X6 is N or CR 10g and; X7 is N or CR 10h and; X8 is N or CR 10i and; X9 is N or CR 10j and; X 10 is N or CR 10k and; Z1 is absent or present, and, if present, is S or SO2; Z2 is absent or present, and, if present, is S or SO2; R 10a is selected from H and alkyl; and R 10b , R 10c , R 10e , R 10f , R 10g , R 10h , R 10i , R 10j , and R 10k are independently selected from H, halogen, and alkyl. It is expressed by:

[0122] In certain embodiments, X aa 11 teeth, [ka] It is expressed by:

[0123] In certain embodiments, X aa 11 teeth, [ka] Represented by TIFF2024525236000020.tif165146.

[0124] In certain embodiments, X aa 11 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; Z1 is absent or present and, if present, is selected from CH2, S, O, NH, SO2, SO2-NH, NH-SO2, NHC(O), and C(O)NH; R 10a is selected from H and alkyl; R 10d is -OH or -L1-L2-L3-R 10d 'And; R 10d ' is -NH2, (C1-C 20 (alkyl)-COH or optionally substituted (C1-C6 alkyl)-aryl; L1 is absent or present, and if present, is a linker; L2 is a linker comprising an ether moiety; L3 is absent or present, and if present, is a linker comprising an amino acid moiety; and R 10b , R 10c , R 10d , R 10e , and R 10f are independently selected from H, halogen, and alkyl. It is expressed by:

[0125] In certain embodiments, R 10d -L1-L2-L3-R 10d ' is.

[0126] In certain embodiments, X aa 11 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; X6 is N or CR 10g and; X7 is N or CR 10h and; X9 is N or CR 10j and; X 10 is N or CR 10k and; Z1 is absent or present and, if present, is selected from CH2, NH, S, SO2, O, SO2-NH, NH-SO2, NHC(O), and C(O)NH; Z2 is absent or present and, if present, is selected from NH, S, SO2, O, SO2-NH, NH-SO2, NHC(O), and C(O)NH; R 10a is selected from H and alkyl; R 10i is -OH or -L1-L2-L3-R 10i 'And; R 10i ' is -NH2, (C1-C 20 (alkyl)-COH or optionally substituted (C1-C6 alkyl)-aryl; L1 is absent or present, and if present, is a linker; L2 is a linker comprising an ether moiety; L3 is absent or present, and if present, is a linker comprising an amino acid moiety; and R 10b , R 10c , R 10e , R 10f , R 10g , R 10h , R 10j , and R 10kare independently selected from H, halogen, and alkyl. It is expressed by:

[0127] In certain embodiments, R 10i -L1-L2-L3-R 10i ' is.

[0128] In certain embodiments, X aa 11 teeth, [ka] [In the formula, X1 is N or CR 10b and; X2 is N or CR 10c and; X4 is N or CR 10e and; X5 is N or CR 10f and; R 10a is selected from H and alkyl; R 10b , R 10c , R 10e , and R 10f is independently selected from H, halogen, and alkyl; and Z3 is a substituted heteroaryl. It is expressed by:

[0129] In certain embodiments, Z3 is a 5-membered substituted heteroaryl.

[0130] In certain embodiments, the 5-membered substituted heteroaryl is a substituted triazolyl.

[0131] In certain embodiments, the substituted heteroaryl is -(C 1~20 It is substituted with -COH.

[0132] In certain embodiments, the substituted heteroaryl is -(C 10~20 It is substituted with -COH.

[0133] In certain embodiments, the substituted heteroaryl is -(C 15 It is substituted with -COH.

[0134] In certain embodiments, X aa 11 teeth, [ka] It is expressed by:

[0135] In certain embodiments, L, when present, is [ka] Selected from TIFF2024525236000026.tif19143.

[0136] In certain embodiments, L2 is [ka] TIFF2024525236000028.tif19140; and each n is independently 1 to 6.

[0137] In certain embodiments, L3, when present, is [ka] The file is TIFF2024525236000030.tif22140.

[0138] In certain embodiments, R 10d ' is (C1~C 15 alkyl)-COH.

[0139] In certain embodiments, R 10d 'teeth, [ka] and R10d '' is a halo.

[0140] In certain embodiments, R 10d '' is I.

[0141] In certain embodiments, R 10i ' is (C1~C 15 alkyl)-COH.

[0142] In certain embodiments, R 10i 'teeth, [ka] and R 10i '' is a halo.

[0143] In certain embodiments, R 10i '' is I.

[0144] In certain embodiments, R 10d teeth, [ka] Selected from TIFF2024525236000034.tif199159TIFF2024525236000035.tif49159.

[0145] In certain embodiments, R 10i teeth, [ka] Selected from TIFF2024525236000037.tif135160.

[0146] In certain embodiments, X aa 10are Phe, Tyr, Trp, homophenylalanine (Hph), homotyrosine (Hty), Bip, α-MeBip, 4-phenyl-3-pyridylalanine, 4-phenyl-4-pyridylalanine, α-MeHph, α-MeTyr, α-MeHty, Tyr(O-phenyl), Phe(4-S-phenyl), Phe(4-SO2-NH-phenyl), Phe(4-CO-NH-phenyl), Cys(S-phenyl), Cys(S-phenyl[2,3,4,5,6-F5]) , Cys(S-phenyl[2,3,4,5-F4]-4-phenyl[2',3',4',5',6'-F5]), Cys(S-phenyl[2,3,4,5-F4]-4-SO2-phenyl[2',3',4',5',6'-F5]), Cy(SO2-NH-phenyl), Dap(3-C[=O]-phenyl), Dap(3-[C=O]-pyridyl), Asp(3-NH-phenyl), and Asp(3-NH-pyridyl).

[0147] In certain embodiments, X aa 11 are Phe, Tyr, Trp, homophenylalanine (Hph), homotyrosine (Hty), Bip, α-MeBip, 4-phenyl-3-pyridylalanine, 4-phenyl-4-pyridylalanine, α-MeHph, α-MeTyr, α-MeHty, Tyr(O-phenyl), Phe(4-S-phenyl), Phe(4-SO2-NH-phenyl), Phe(4-CO-NH-phenyl), Cys(S-phenyl), Cys(S-phenyl[2,3,4,5,6-F5]) , Cys(S-phenyl[2,3,4,5-F4]-4-phenyl[2',3',4',5',6'-F5]), Cys(S-phenyl[2,3,4,5-F4]-4-SO2-phenyl[2',3',4',5',6'-F5]), Cy(SO2-NH-phenyl), Dap(3-C[=O]-phenyl), Dap(3-[C=O]-pyridyl), Asp(3-NH-phenyl), and Asp(3-NH-pyridyl).

[0148] In certain embodiments, R XNis -N(Rx)2, where each Rx is H.

[0149] In certain embodiments, R XN is defined as -N(Rx)2, where one occurrence of Rx is hydrogen and the other occurrence is an amino acid residue X aa 0 [It is].

[0150] In certain embodiments, R YC is -C(O)(NR Y )2[where each R Y is H].

[0151] In certain embodiments, R YC is -C(O)(NR Y )2; R Y One occurrence of is hydrogen and the other is R Y The other occurrence of is -L4-L5-L6-L7-R Y ' and R Y ' is (C1~C 20 (alkyl)-COH or optionally substituted (C1-C6 alkyl)-aryl; L4 is absent or present, and if present, is a linker comprising an amino acid moiety; L5 is absent or present, and if present, is a linker comprising an amino acid moiety; L6 is absent or present, and if present, is a linker comprising an ether moiety; and L7 is a linker that comprises an amino acid moiety.

[0152] In certain embodiments, L4 is present, [ka] and m is 1 to 6.

[0153] In certain embodiments, L5 is present, [ka] and l is 1 to 6.

[0154] In certain embodiments, L5 is present, [ka] and n is 1 to 6.

[0155] In certain embodiments, L6 is [ka] The file is TIFF2024525236000042.tif16141.

[0156] In certain embodiments, R Y ' is (C 10 ~C 16 alkyl)-COH.

[0157] In certain embodiments, R Y 'teeth, [ka] and R Y '' is a halo.

[0158] In certain embodiments, R Y '' is I.

[0159] In certain embodiments, R Y One occurrence of is hydrogen and the other occurrence of is [ka] Selected from TIFF2024525236000045.tif103160.

[0160] In certain embodiments, the polypeptide is selected from the polypeptides listed in Table 4.

[0161] In certain embodiments, the polypeptide is selected from the polypeptides listed in Table 5.

[0162] In certain embodiments, the polypeptide is selected from the polypeptides listed in Table 6.

[0163] In certain embodiments, the polypeptide comprises: [ka] Represented by TIFF2024525236000047.tif118138.

[0164] In certain embodiments, the polypeptide comprises: [ka] TIFF2024525236000049.tif159166[in the formula, each n is independently 10 to 20 (e.g., 15); Each R 2A is independently -H or alkyl; and Each R 10A are independently -H or alkyl. It is expressed by:

[0165] In certain embodiments, each n is independently 10 to 20 (e.g., 15); 2A is independently -H or -CH; and each R 10A is independently -H or -CH3.

[0166] In certain embodiments, the polypeptide comprises: [ka] [In the formula, each m is independently 1 to 10; Each R 2A is independently -H or alkyl; Each R 10Ais independently -H or alkyl; Each R 10Z is independently halo (e.g., I); and Each R 11Z are independently halo (e.g., I). It is expressed by:

[0167] In certain embodiments, each n is independently 1 to 10; 2A is independently -H or -CH; each R 10A is independently -H or -CH; each R 10Z is independently halo (e.g., I); and each R 1aZ is independently halo (e.g., I).

[0168] In certain embodiments, each R 10Z is I; and each R 11Z is I.

[0169] In certain embodiments, the polypeptide comprises: [ka] It is expressed by:

[0170] In certain embodiments, the polypeptide comprises: [ka] It is expressed by:

[0171] In certain embodiments, the polypeptide comprises: [ka] Represented by TIFF2024525236000054.tif41158.

[0172] In certain embodiments, the polypeptide comprises: [ka] Represented by TIFF2024525236000056.tif49163.

[0173] In certain embodiments, S-aryl 1 has the structure: [ka] has.

[0174] In certain embodiments, S-aryl 2 has the structure: [ka] has.

[0175] In certain embodiments, S-aryl 3 has the structure: [ka] has.

[0176] In certain embodiments, R 10d or R 10i can bind to hydroxyl, amino, carboxyl, azido, alkynyl, or methyl groups substituted with thiol, hydroxyl, amino, carboxyl, azido, or alkynyl, as well as to serum albumin (fatty acids (e.g., C 16 Or C 18 ), in the manner exemplified by GLP-1 peptide agonists such as liraglutide and semaglutide (17-19), or further by further conjugation via such substituents (e.g., thioethers, ethers, amines, esters, triazines) to functional groups that enhance the pharmacokinetic properties by incorporation of phenyl iodide (20) and other known serum albumin binding moieties represented by aryl halides, as exemplified below.

[0177] In certain embodiments, R 10d or R 10i teeth, [ka] Selected from TIFF2024525236000061.tif94159.

[0178] In certain embodiments, R YC is -C(O)NHR Y [In the formula, NHR Y teeth, [ka] TIFF2024525236000063.tif145162].

[0179] In certain embodiments, R YC is -C(O)NHR Y [In the formula, NHR Y teeth, [ka] TIFF2024525236000065.tif182152TIFF2024525236000066.tif177153TIFF2024525236000067.tif68147].

[0180] In certain embodiments, R YC is -C(O)NHR Y [In the formula, NHR Y teeth, [ka] TIFF2024525236000069.tif188163].

[0181] In certain embodiments, X aa 10are Phe, Tyr, Trp, homophenylalanine (Hph), homotyrosine (Hty), Bip, α-MeBip, 4-phenyl-3-pyridylalanine, 4-phenyl-4-pyridylalanine, α-MeHph, α-MeTyr, α-MeHty, Tyr(O-phenyl), Phe(4-S-phenyl), Phe(4-SO2-NH-phenyl), Phe(4-CO-NH-phenyl), and Cys(S-phenyl), Cys(S-phenyl[2,3,4,5,6-F]), Cys(S-phenyl[2,3,4,5-F]-4-phenyl[2',3',4',5',6'-F]), and Cys(S-phenyl[2,3,4,5-F]-4-SO-phenyl[2',3',4',5',6'-F]).

[0182] In certain embodiments, X aa 10 is optionally substituted Cy(SO2-NH-phenyl), i.e., the -SO3H group of cysteic acid is replaced with -SO2-NH-phenyl.

[0183] In certain embodiments, X aa 10 is an optionally substituted amino acid residue selected from Dap(3-C[=O]-phenyl) (i.e., the amino group of the side chain is substituted with -NHC(O)-phenyl), Dap(3-[C=O]-pyridyl) (i.e., the amino group of the side chain is substituted with -NHC(O)-4-pyridinyl), Asp(3-NH-phenyl) (i.e., the carboxyl group of the side chain is substituted with -C(O)NH-phenyl), and Asp(3-NH-pyridyl) (i.e., the carboxyl group of the side chain is substituted with -C(O)NH-4-pyridinyl).

[0184] In certain embodiments, X aa 11 is optionally substituted Cy(SO2-NH-phenyl), i.e., the -SO3H group of cysteic acid is replaced with -SO2-NH-phenyl.

[0185] In certain embodiments, X aa 11 is an optionally substituted amino acid residue selected from Dap(3-C[=O]-phenyl) (i.e., the amino group of the side chain is substituted with -NHC(O)-phenyl), Dap(3-[C=O]-pyridyl) (i.e., the amino group of the side chain is substituted with -NHC(O)-4-pyridinyl), Asp(3-NH-phenyl) (i.e., the carboxyl group of the side chain is substituted with -C(O)NH-phenyl), and Asp(3-NH-pyridyl) (i.e., the carboxyl group of the side chain is substituted with -C(O)NH-4-pyridinyl).

[0186] In certain embodiments, X aa 2 and / or X aa 11 is a D-amino acid residue.

[0187] In certain embodiments, X aa 1 ~X aa 11 Each of is an L-amino acid residue.

[0188] In certain embodiments, the amino acid residues, when substituted, are substituted with alkyl or halo.

[0189] In certain embodiments, the amino acid residues, when substituted, are substituted with alkyl, hydroxyl, or halo.

[0190] In certain embodiments, any one of the amino acid residues is selected from the naturally occurring amino acids.

[0191] In certain embodiments, any one of the amino acid residues is selected from a non-naturally occurring amino acid.

[0192] In certain embodiments, the compounds are atropisomers. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, hydrogen can be replaced with deuterium or tritium, or carbon can be replaced with an isotopically enriched atom. 13 C- or 14 Compounds produced by substituting C-enriched carbons are within the scope of the invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the invention. For example, the variable R 1 In the case of, the (C1-C4) alkyl or -O-(C1-C4) alkyl can be suitably deuterated (eg, -CD3, -OCD3).

[0193] Any of the compounds of the present invention can also be radiolabeled for the preparation of radiopharmaceuticals.

[0194] Treatment method One aspect of the invention relates to a method for treating or preventing a disease or disorder mediated at least in part by glucagon-like peptide 1 in a subject in need of such treatment or prevention, comprising the step of administering to the subject an effective amount of a polypeptide of sequence (I).

[0195] In certain embodiments, a method for treating or preventing diabetes in a subject in need thereof, comprising administering to the subject an effective amount of a polypeptide of sequence (I).

[0196] In certain embodiments, the diabetes is type II diabetes.

[0197] In certain embodiments, methods for treating, preventing, or delaying the onset of complications associated with diabetes, such as macrovascular complications and microvascular complications, e.g., retinopathy, neuropathy, nephropathy, and delayed wound healing, and related diseases, such as insulin resistance (impaired glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, hyperlipidemia, e.g., hypertriglyceridemia, Syndrome X, atherosclerosis, and hypertension, and methods for increasing high density lipoprotein levels.

[0198] In certain embodiments, the method further comprises administering an antidiabetic agent.

[0199] In certain embodiments, the method further comprises administering a lipid-lowering agent, which may be applied in the context of human immunodeficiency virus (HIV) and its treatment.

[0200] In certain embodiments, a method for treating or preventing obesity or an associated metabolic disorder, such as polycystic ovarian disease (PCOS), in a subject in need thereof, comprising administering to the subject an effective amount of a polypeptide of sequence (I).

[0201] In certain embodiments, the method further comprises administering an anti-obesity agent.

[0202] In certain embodiments, a method for treating or preventing cardiovascular disease in a subject in need thereof, comprising administering to the subject an effective amount of a polypeptide of sequence (I).

[0203] In certain embodiments, the method further comprises administering an antihypertensive agent.

[0204] In certain embodiments, a method for treating or preventing a neurodegenerative disease in a subject in need thereof, comprising administering to the subject an effective amount of a polypeptide of sequence (I).

[0205] In certain embodiments, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, Huntington's disease, and prion diseases.

[0206] In certain embodiments, a method for treating or preventing traumatic brain injury (TBI) in a subject in need thereof, comprising administering to the subject an effective amount of a polypeptide of sequence (I).

[0207] In certain embodiments, a method for treating or preventing non-alcoholic steatohepatitis (NASH) in a subject in need thereof, comprising administering to the subject an effective amount of a polypeptide of sequence (I).

[0208] In some embodiments of any one of the disclosed methods, the polypeptide of sequence (I) is R XN -X aa 1 -X aa 2 -X aa 3 -X aa 4 -X aa 5 -X aa 6 -X aa 7 -X aa 8 -X aa 9 -X aa 10 -X aa 11 -R YC [In the formula, R XNis H (i.e., des-amino) and -N(Rx)2, where Rx is, independently for each occurrence, H or optionally substituted alkyl, arylalkyl, heteroarylalkyl, formyl, acetyl, alkanoyl, -C(O)-alkyloxy, -C(O)-aryloxy, -C(O)-aralkyloxy, -C(O)-heterocyclyloxy, -C(O)-heteroarylalkyloxy, -C(O)NH-alkyl, -C(O)NH-aryl, -C(O)NH-arylalkyl, -SO2-heterocyclyl, -SO2-alkyl, -SO2-aryl, -SO2-arylalkyl, -SO2-heteroarylalkyl, -SO2-heteroaryl, or ureido; or one occurrence of Rx is hydrogen and the other occurrence is an amino acid residue X aa 0 X is selected from aa 1 is the N-terminal group of; X aa 0 is an optionally substituted amino acid residue selected from Gly, Pro, Arg, Glu, His, Phe, and Trp; X aa 1 is an optionally substituted amino acid residue containing an amino acid side chain containing alkyl, aryl, or heteroaryl; X aa 2 is an optionally substituted amino acid residue selected from Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; X aa 3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 4 is an amino acid residue selected from Gly, Ala, Aib, and β-Ala; X aa 5is an optionally substituted amino acid selected from Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; X aa 6 is an optionally substituted amino acid residue disubstituted at the alpha carbon, provided that one of the substituents is an optionally substituted aryl or heteroaryl; X aa 7 is an optionally substituted amino acid residue containing a hydroxyl-containing amino acid side chain; X aa 8 is an optionally substituted amino acid residue selected from Ser, His, and Asn; X aa 9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 10 is an optionally substituted amino acid residue containing a sulfide and / or an optionally substituted aryl or heteroaryl containing amino acid side chain; X aa 11 is an optionally substituted amino acid residue containing a sulfide and / or an optionally substituted aryl or heteroaryl containing amino acid side chain; and R YC is the structure -C(O)N(R Y )2(wherein, R Y is, independently for each occurrence, hydrogen or a PK modifying group; aa 11 is the C-terminal group of It is defined as follows.

[0209] Pharmaceutical Compositions, Routes of Administration, and Dosing In certain embodiments, the present invention is directed to a pharmaceutical composition comprising a compound of the present invention, i.e., a polypeptide, and a pharma- ceutically acceptable carrier. In certain embodiments, the pharmaceutical composition comprises a plurality of compounds of the present invention and a pharma- ceutically acceptable carrier.

[0210] In certain embodiments, the pharmaceutical composition of the present invention further comprises at least one additional pharma- ceutical active agent other than the compound of the present invention. The at least one additional pharma- ceutical active agent may be, for example, an agent useful in the treatment of diabetes.

[0211] Pharmaceutical compositions of the present invention can be prepared by combining one or more compounds of the present invention with a pharma- ceutically acceptable carrier and, optionally, one or more additional pharma- ceutical active agents.

[0212] As stated above, "effective amount" refers to any amount sufficient to achieve a desired biological effect. In combination with the teachings provided herein, by selecting from among various active compounds and considering factors such as potency, relative bioavailability, patient weight, severity of adverse side effects, and method of administration, one can design an effective prophylactic or therapeutic treatment regimen that does not cause substantial undesirable toxicity and is effective in treating a particular subject. The effective amount for any particular application may vary depending on factors such as the disease or condition being treated, the particular compound of the invention being administered, the size of the subject, or the severity of the disease or condition. Those skilled in the art can empirically determine the effective amount of a particular compound of the invention and / or other therapeutic agent without necessitating undue experimentation. A maximum dose, i.e., the highest dose that is safe according to any medical judgment, may be used. Multiple administrations per day may be contemplated to achieve an appropriate systemic level of the compound. An appropriate systemic level may be determined, for example, by measuring the patient's peak or sustained plasma levels of the drug. "Dose" and "dosage" are used interchangeably herein.

[0213] In certain embodiments, intravenous administration of the compound may typically be from 0.1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 0.1 mg / kg / day to 2 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 0.5 mg / kg / day to 5 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 1 mg / kg / day to 20 mg / kg / day. In one embodiment, intravenous administration of the compound may typically be from 1 mg / kg / day to 10 mg / kg / day.

[0214] In general, the daily oral dose of the compound is about 0.01 milligrams / kg per day to 1000 milligrams / kg per day for human subjects. Oral doses ranging from 0.5 to 50 milligrams / kg in one or more administrations per day are expected to provide therapeutic results. Dosage may be appropriately adjusted to achieve desired local or systemic drug levels, depending on the method of administration. For example, intravenous administration is expected to be one to several orders of magnitude lower per day. If the response in the subject is inadequate at such doses, even higher doses (or higher effective doses by a different, more localized delivery route) may be used, to the extent that patient tolerance permits. Multiple administrations per day are contemplated to achieve adequate systemic levels of the compound.

[0215] For any compound described herein, the therapeutically effective amount can be determined first from animal models. The therapeutically effective dose can also be determined from human data for compounds tested in humans and for compounds known to exhibit similar pharmacological activity, such as other related active agents. Higher doses may be required for parenteral administration. The applied dose can be adjusted based on the relative bioavailability and efficacy of the administered compound. It is well within the capabilities of a person skilled in the art to adjust the dose to achieve maximum efficacy based on the above methods and other methods known in the art.

[0216] The formulations of the present invention may be administered in pharma- ceutically acceptable solutions, which may routinely contain pharma- ceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.

[0217] For use in therapy, an effective amount of the compound can be administered to a subject by any method that delivers the compound to the desired surface.The administration of pharmaceutical compositions can be achieved by any means known to those skilled in the art.Routes of administration include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (bladder), oral, subcutaneous, direct injection (e.g., into tumor or abscess), mucosal (e.g., topical to eye), inhalation, and topical.

[0218] For intravenous and other parenteral routes of administration, the compounds of the present invention can be formulated as lyophilized preparations, as lyophilized preparations of liposome-entrapped or encapsulated active compounds, as lipid complexes in aqueous suspension, or as salt complexes. Lyophilized preparations are generally reconstituted in a suitable aqueous solution, such as sterile water or saline, immediately prior to administration.

[0219] For oral administration, the compounds can be easily formulated by combining the active compound(s) with pharma- ceutically acceptable carriers well known in the art. Such carriers allow the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like for oral ingestion by the subject to be treated. Pharmaceutical preparations for oral use can be obtained by grinding the resulting mixture and processing the mixture of granules, optionally after adding suitable auxiliary agents, to obtain tablets or dragee cores, as a solid excipient. Suitable excipients are, in particular, fillers, such as sugars, such as lactose, sucrose, mannitol, or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate. Optionally, the oral preparations may also be formulated in saline or buffer, such as EDTA for neutralizing internal acidic conditions, or may be administered without any carrier.

[0220] Oral dosage forms of the above component(s) are also specifically contemplated. The component(s) may be chemically modified to effect oral delivery of the derivative. In general, the contemplated chemical modification is the attachment of at least one moiety to the component molecule itself, said moiety (a) inhibiting acid hydrolysis; and (b) allowing uptake from the stomach or intestine into the bloodstream. Also desirable is an increase in the overall stability of the component(s) and an increase in circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, and polyproline. Abuchowski and Davis, "Soluble Polymer-Enzyme Adducts", In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, NY, pp. 367-383 (1981); Newmark et al., J Appl Biochem 4:185-9 (1982). Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane. For pharmaceutical applications, as indicated above, polyethylene glycol moieties are preferred.

[0221] For the component (or derivative), the location of release may be the stomach, the small intestine (the duodenum, jejunum, or ileum), or the large intestine. Those skilled in the art have formulations available that will not dissolve in the stomach, but will release the material in the duodenum or elsewhere in the intestine. Preferably, the release avoids the deleterious effects of the stomach environment, either by protection of the compound (or derivative) of the invention or by release of the bioactive material beyond the stomach environment, such as in the intestine.

[0222] To ensure full gastric resistance, a coating impermeable to at least pH 5.0 is essential. Examples of the more common inactive ingredients used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings may also be used as mixed coatings.

[0223] A coating or mixture of coatings may also be used on tablets that are not intended for protection against the stomach. This may include sugar coatings, or coatings that make the tablet easier to swallow. Capsules may consist of a hard shell (e.g., gelatin) for delivery of dry therapeutics (e.g., powder); for liquid forms, a soft gelatin shell may be used. The shell material for cachets may be thick starch or other edible paper. For pills, lozenges, molded tablets, or powder tablets, moist massing techniques may be used.

[0224] The therapeutic agent may be included in the formulation as fine multiparticulates in the form of granules or pellets about 1 mm in size. The formulation of the material for capsule administration may also be as a powder, lightly compressed plugs, or even as tablets. The therapeutic agent may be prepared by compression.

[0225] Colorants and flavoring agents may all be included. For example, the compounds (or derivatives) of the present invention may be formulated (e.g., by liposomal or microsphere encapsulation) and then further included within an edible product, such as a refrigerated beverage, containing colorants and flavoring agents.

[0226] Inert materials may be used to dilute or increase the volume of the therapeutic agent. These diluents include carbohydrates, especially mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans, and starch. Certain inorganic salts, such as calcium triphosphate, magnesium carbonate, and sodium chloride, may also be used as bulking agents. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress, and Avicell.

[0227] Disintegrants may be included when the therapeutic agent is formulated into a solid dosage form. Materials used as disintegrants include, but are not limited to, starch, including Explotab, a commercially available disintegrant based on starch. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid-type carboxymethylcellulose, natural sponge and bentonite may all be used. Another form of disintegrant is the insoluble cation exchange resin. Powdered gums may be used as disintegrants and as binders, and these may include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.

[0228] Binders may be used to bind the therapeutic agent together to form a hard tablet, and include naturally derived materials such as acacia, tragacanth, starch, and gelatin. Others include methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Both polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) can be used in alcoholic solutions to granulate the therapeutic agent.

[0229] Antifriction agents may be included in the formulation of the therapeutic agent to prevent sticking during the formulation process. Lubricants may be used as a layer between the therapeutic agent and the die wall, including but not limited to stearic acid, such as its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants may also be used, such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, Carbowax 4000 and 6000.

[0230] Glidants may be added which may improve the flow properties of the drug during formulation and aid in rearrangement during compression. Glidants include starch, talc, pyrogenic silica and hydrated silicoaluminate.

[0231] Surfactants may be added as wetting agents to aid in dissolving therapeutic agents in aqueous environments.Surfactants include anionic surfactants such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate.Cationic surfactants that can be used include benzalkonium chloride and benzethonium chloride.Possible nonionic surfactants that can be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose.These surfactants can be present alone or as a mixture in various ratios in the formulation of the compound or derivative of the present invention.

[0232] Orally usable pharmaceutical preparations include push-fit capsules made of gelatin, as well as soft sealed capsules made of gelatin and plasticizers, such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient mixed with a filler, such as lactose, a binder, such as starch, and / or a lubricant, such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid, such as fatty oils, liquid paraffin, or liquid polyethylene glycol. Additionally, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres are well defined in the art. All formulations for oral administration should be in dosages suitable for such administration.

[0233] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0234] For local administration, the compounds may be formulated as solutions, gels, ointments, creams, suspensions, etc., as is well known in the art. Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal, oral or pulmonary administration.

[0235] For administration by inhalation, the compounds for use according to the present invention may be conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges, for example of gelatin, for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, for example lactose or starch.

[0236] Pulmonary delivery of the compounds (or salts thereof) disclosed herein is also contemplated herein. The compounds are delivered to the lungs of a mammal during inhalation and travel across the pulmonary epithelial lining into the bloodstream. Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990)(leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl. 5):143-146 (1989)(endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989)(α1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146(α-1-proteinase); Oswein et al., 1990, “Aerosolization of Proteins”, Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March, (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-gamma and tumor necrosis factor alpha) and Platz et al., U.S. Patent No. 5,284,656 (granulocyte colony stimulating factor; incorporated by reference). Methods and compositions for pulmonary delivery of drugs for systemic effect are described in U.S. Patent No. 5,451,569, issued September 19, 1995 to Wong et al. (incorporated by reference).

[0237] A wide variety of mechanical devices designed for pulmonary delivery of therapeutic products are contemplated for use in the practice of the present invention, including, but not limited to, nebulizers, metered dose inhalers, and dry powder inhalers, all of which are well known to those skilled in the art.

[0238] Some specific examples of commercially available devices suitable for the practice of the present invention are the Ultravent nebulizer manufactured by Mallinckrodt, Inc., St. Louis, Mo.; the Acorn II nebulizer manufactured by Marquest Medical Products, Englewood, Colo.; the Ventolin metered dose inhaler manufactured by Glaxo Inc., Research Triangle Park, North Carolina; and the Spinhaler powder inhaler manufactured by Fisons Corp., Bedford, Mass.

[0239] All such devices require the use of suitable formulations for dispensing the compounds of the present invention. Typically, each formulation is specific to the type of device used, and may include the use of suitable propellant materials in addition to the usual diluents, adjuvants and / or carriers useful in therapy. Also contemplated is the use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers. Chemically modified compounds of the present invention can also be prepared into various formulations, depending on the type of chemical modification or the type of device used.

[0240] Formulations suitable for use with jet or ultrasonic nebulizers typically contain the compound (or derivative) of the invention dissolved in water at a concentration of about 0.1-25 mg of biologically active compound of the invention per mL of solution. The formulation may also include a buffer and a simple sugar (e.g., for inhibitor stabilization and regulation of osmotic pressure). Nebulizer formulations may also contain a surfactant to reduce or prevent surface-induced aggregation of the compound of the invention caused by atomization of the solution when forming an aerosol.

[0241] The formulation for use in metered dose inhalation devices generally comprises a finely divided powder containing the compound (or derivative) of the present invention suspended in a propellant with the aid of a surfactant.The propellant can be any conventional material used for this purpose, such as chlorofluorocarbons, hydrochlorofluorocarbons, hydrofluorocarbons, or hydrocarbons, such as trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or combinations thereof.Suitable surfactants include sorbitan trioleate and soybean lecithin.Oleic acid can also be useful as a surfactant.

[0242] Formulations for dispensing from a powder inhalation device comprise a finely divided dry powder containing the compound (or derivative) of the invention and may also contain a bulking agent, e.g., lactose, sorbitol, sucrose, or mannitol, in an amount to facilitate dispersion of the powder from the device, e.g., 50-90% by weight of the formulation. The compound (or derivative) of the invention should advantageously be prepared in a particulate form having a mean particle size of less than 10 micrometers (μm), most preferably 0.5-5 μm, for most effective delivery to the deep lung.

[0243] Nasal delivery of the pharmaceutical compositions of the invention is also contemplated. Nasal delivery allows for passage of the pharmaceutical compositions of the invention directly into the bloodstream after administration of the therapeutic product to the nose, without the need for deposition of the product in the lungs. Formulations for nasal delivery include those using dextran or cyclodextran.

[0244] For nasal administration, a useful device is a small hard bottle with a metered dose sprayer attached. In one embodiment, the metered dose is delivered by drawing the pharmaceutical composition of the present invention in solution into a chamber of a specified volume, which has an opening dimensioned to aerosolize the aerosol formulation by forming a spray when the liquid in the chamber is compressed. The chamber is compressed to administer the pharmaceutical composition of the present invention. In a specific embodiment, the chamber is a piston mechanism. Such devices are commercially available.

[0245] Alternatively, a plastic squeeze bottle is used with an opening or aperture sized to aerosolize the aerosol formulation by forming a spray when squeezed. The aperture is usually found at the top end of the bottle, which is generally tapered to partially fit into the nostril for efficient administration of the aerosol formulation. Preferably, the nasal inhaler provides a metered amount of the aerosol formulation for administration of a metered dose of the drug.

[0246] The compounds may be formulated for parenteral administration by injection, for example, by bolus injection or continuous infusion, when it is desired to deliver the compounds systemically. The formulations for injection may be presented in unit dosage form, for example, in ampoules or multi-dose containers, with added preservatives. The compositions may take the form of suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents, such as suspending, stabilizing and / or dispersing agents.

[0247] Pharmaceutical preparations for parenteral administration include aqueous solutions of active compounds in water-soluble form. In addition, suspensions of active compounds may be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compound, allowing the preparation of highly concentrated solutions.

[0248] Alternatively, the active compounds may be in powder form for constitution with a suitable vehicle, eg, sterile pyrogen-free water, before use.

[0249] The compounds may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, eg, containing conventional suppository bases such as cocoa butter or other glycerides.

[0250] In addition to the above formulations, the compounds may also be formulated as depot preparations. Such long-acting preparations may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example as a sparingly soluble salt.

[0251] The pharmaceutical compositions may also comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[0252] Suitable liquid or solid pharmaceutical formulation forms are, for example, aqueous or saline solutions for inhalation, microencapsulated forms, cochleated forms, coated on fine gold particles, encapsulated in liposomes, nebulized forms, aerosols, pellets for skin implantation, or dry forms on sharps for scratching the skin. Pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or preparations in which active compounds are sustainedly released, in which excipients and additives, and / or auxiliary agents, such as disintegrants, binders, coating agents, swelling agents, lubricants, flavoring agents, sweetening agents or solubilizers, are conventionally used as described above. Pharmaceutical compositions are suitable for use in various drug delivery systems. For a brief overview of methods for drug delivery, see Langer R, Science 249:1527-33 (1990).

[0253] The compounds of the present invention and optionally other therapeutic agents may be administered per se (neat) or in the form of a pharma- ceutically acceptable salt or co-crystal. When used in medicine, the salt or co-crystal should be pharma- ceutically acceptable, but pharma- ceutically unacceptable salts or co-crystals may be conveniently used to prepare pharma- ceutically acceptable salts or co-crystals thereof. Such salts include, but are not limited to, those prepared from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, p-toluenesulfonic acid, tartaric acid, citric acid, methanesulfonic acid, formic acid, malonic acid, succinic acid, naphthalene-2-sulfonic acid, and benzenesulfonic acid. Such salts may also be prepared as alkali metal or alkaline earth salts, such as sodium, potassium, or calcium salts of the carboxylic acid group.

[0254] Suitable buffering agents include acetic acid and salts (1-2% w / v), citric acid and salts (1-3% w / v), boric acid and salts (0.5-2.5% w / v), and phosphoric acid and salts (0.8-2% w / v). Suitable preservatives include benzalkonium chloride (0.003-0.03% w / v), chlorobutanol (0.3-0.9% w / v), parabens (0.01-0.25% w / v), and thimerosal (0.004-0.02% w / v).

[0255] The pharmaceutical composition of the present invention contains an effective amount of the compound described herein and optionally a therapeutic agent, contained in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers, diluents or encapsulating substances suitable for administration to humans or other vertebrates. The term "carrier" means a natural or synthetic organic or inorganic component with which the active ingredient is combined to facilitate application. The components of the pharmaceutical composition are also compatible with the compound of the present invention and with each other in a manner that there is no interaction that would substantially impair the desired pharmaceutical efficiency.

[0256] The therapeutic agent(s), including but not limited to the compounds of the present invention, may be provided in particles. Particles, as used herein, refers to nanoparticles or microparticles (or possibly larger particles), which may consist in whole or in part of the compounds of the present invention or other therapeutic agent(s) described herein. The particles may contain the therapeutic agent(s) in a core surrounded by a coating, including but not limited to an enteric coating. The therapeutic agent(s) may also be dispersed throughout the particle. The therapeutic agent(s) may also be adsorbed to the particle. The particles may have any order of release kinetics, such as zero order release, first order release, second order release, delayed release, sustained release, immediate release, and any combination thereof. The particles may contain, in addition to the therapeutic agent(s), any of the materials routinely used in the pharmaceutical and medical fields, including but not limited to erodible, non-erodible, biodegradable, or non-biodegradable materials or combinations thereof. The particles may be microcapsules containing the compounds of the present invention in solution or in a semi-solid state. The particles may be of virtually any shape.

[0257] Both non-biodegradable and biodegradable polymeric materials can be used to manufacture particles for delivery of therapeutic agent(s). Such polymers can be natural or synthetic. The polymer is selected based on the period of time over which release is desired. Bioadhesive polymers of particular interest include the bioerodible hydrogels described in Sawhney HS et al. (1993) Macromolecules 26:581-7, the teachings of which are incorporated herein. These include polyhyaluronic acid, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginates, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0258] The therapeutic agent(s) may be included in a controlled release system. The term "controlled release" is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation is controlled. It refers to immediate release formulations and non-immediate release formulations, which include, but are not limited to, sustained release formulations and delayed release formulations. The term "sustained release" (also referred to as "extended release") is used in the conventional sense to refer to a drug formulation that provides a gradual release of drug over an extended period of time, preferably, but not necessarily, providing a substantially constant blood level of drug over an extended period of time. The term "delayed release" is used in the conventional sense to refer to a drug formulation in which there is a time delay between administration of the formulation and the release of drug from the formulation. "Delayed release" may or may not include a gradual release of drug over an extended period of time, and therefore may or may not be a "sustained release".

[0259] The use of long-term sustained release implants may be particularly suitable for treating chronic conditions. "Long-term" release, as used herein, means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, and preferably 30 to 60 days. Long-term sustained release implants are well known to those skilled in the art and include some of the release systems described above.

[0260] combination The present invention includes within its scope pharmaceutical compositions comprising as an active ingredient a therapeutically effective amount of at least one of the polypeptides of sequence I, alone or in combination with a pharmaceutical carrier or diluent. Optionally, the polypeptides of the present invention can be used in any one of the disclosed methods, alone, in combination with other compounds of the present invention, or in combination with one or more other therapeutic agents disclosed herein, such as antidiabetic agents or other pharma- ceutical active materials.

[0261] The polypeptides of the present invention may be used in combination with other GLP-1 receptor modulators (e.g., agonists or partial agonists, such as peptide agonists) or other suitable therapeutic agents useful in the treatment of the aforementioned disorders, including: antidiabetic agents; antihyperglycemic agents; hypolipidemic / lipid lowering agents; antiobesity agents (including appetite suppressants / regulators) and antihypertensive agents. Additionally, the compounds of the present invention may be combined with one or more of the following therapeutic agents; infertility agents, agents for treating polycystic ovary syndrome, agents for treating growth disorders, agents for treating frailty, agents for treating arthritis, agents for preventing allograft rejection in transplantation, agents for treating autoimmune diseases, anti-AIDS agents, anti-osteoporosis agents, agents for treating immunomodulatory disorders, antithrombotic agents, agents for treating cardiovascular diseases, antibiotics, antipsychotic agents, agents for treating chronic inflammatory bowel disease or syndrome, and / or agents for treating anorexia nervosa.

[0262] Examples of suitable antidiabetic agents for use in combination with the compounds of the invention include biguanides (e.g., metformin or phenformin), glucosidase inhibitors (e.g., acarbose or miglitol), insulins (including insulin secretagogues or insulin sensitizers), meglitinides (e.g., repaglinide), sulfonylureas (e.g., glimepiride, glyburide, gliclazide, chlorpropamide, and glipizide), biguanide / glyburide combinations (e.g., Glucovance®), thiazolidinediones (e.g., troglitazone, rosiglitazone, and pioglitazone), PPAR-alpha agonists, PPAR-gamma agonists, PPAR alpha / gamma dual agonists, glycogen phosphorylase inhibitors, inhibitors of fatty acid binding protein (aP2), DPP-IV inhibitors, and SGLT2 inhibitors.

[0263] Other suitable thiazolidinediones include Mitsubishi's MCC-555 (disclosed in U.S. Pat. No. 5,594,016), Glaxo-Welcome's GL-262570, englitazone (CP-68722, Pfizer) or darglitazone (CP-86325, Pfizer), isaglitazone (MIT / J&J), JTT-501 (JPNT / P&U), L-895645 (Merck), R-119702 (Sankyo / WL), NN-2344 (Dr. Reddy / NN), or YM-440 (Yamanouchi).

[0264] Suitable PPAR alpha / gamma dual agonists include muraglitazar (Bristol-Myers Squibb), AR-HO39242 (Astra / Zeneca), GW-409544 (Glaxo-Wellcome), KRP297 (Kyorin Merck), and Murakami et al, “A Novel Insulin Sensitizer Acts as a Coligand for Peroxisome Proliferation-Activated Receptor Alpha (PPAR alpha) and PPAR gamma. Effect on PPAR alpha Activation on Abnormal Lipid Metabolism in Liver of Zucker Fatty Rats”, Diabetes 47, 1841-1847. (1998), and U.S. patent application Ser. No. 09 / 644,598, filed Sep. 18, 2000, the disclosure of which is incorporated herein by reference (using the dosages set forth therein, those compounds designated as preferred are preferred for use herein).

[0265] Suitable aP2 inhibitors include those disclosed in U.S. patent application Ser. No. 09 / 391,053, filed Sep. 7, 1999, and U.S. patent application Ser. No. 09 / 519,079, filed Mar. 6, 2000 (using the dosages described herein).

[0266] Suitable DPP4 inhibitors which may be used in combination with the compounds of the invention include those disclosed in WO 99 / 38501, WO 99 / 46272, WO 99 / 67279 (PROBIODRUG), WO 99 / 67278 (PROBIODRUG), WO 99 / 61431 (PROBIODRUG), NVP-DPP728A (1-[[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(S)-pyrrolidine) (Novartis), LAF237, saxagliptin, MK0431, TSL-225 (tryptophyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) (Yamada et al., Biochemistry, 38(36), 11597-11603, 1999). al, Bioorg. & Med. Chem. Lett. 8 (1998) 1537-1540), the 2-cyanopyrrolidides and 4-cyanopyrrolidides disclosed in Ashworth et al, Bioorg. & Med. Chem. Lett., Vol. 6, No. 22, pp 1163-1166 and 2745-2748 (1996) (using dosages as set forth in the above references).

[0267] Suitable meglitinides include nateglinide (Novartis) or KAD1229 (PF / Kissei).

[0268] Examples of other suitable glucagon-like peptide-1 (GLP-1) compounds that may be used in combination with the GLP-1 receptor modulators (e.g., agonists or partial agonists) of the present invention include GLP-1(1-36)amide, GLP-1(7-36)amide, GLP-1(7-37) (disclosed in U.S. Pat. No. 5,614,492 to Habener), as well as AC2993 (Amylin), LY-315902 (Lilly), and NN2211 (Novo Nordisk).

[0269] Examples of suitable hypolipidemic / lipid lowering agents for use in combination with the compounds of the present invention include one or more of MTP inhibitors, HMG CoA reductase inhibitors, squalene synthetase inhibitors, fibric acid derivatives, ACAT inhibitors, lipoxygenase inhibitors, cholesterol absorption inhibitors, ileal Na + / bile acid cotransporter inhibitors, upregulators of LDL receptor activity, bile acid sequestrants, cholesterol ester transfer protein inhibitors (e.g., CP-529414 (Pfizer)) and / or nicotinic acid and its derivatives.

[0270] MTP inhibitors that may be used as described above include those disclosed in U.S. Patent No. 5,595,872, U.S. Patent No. 5,739,135, U.S. Patent No. 5,712,279, U.S. Patent No. 5,760,246, U.S. Patent No. 5,827,875, U.S. Patent No. 5,885,983 and U.S. Patent No. 5,962,440, all of which are incorporated herein by reference.

[0271] HMG CoA reductase inhibitors that may be used in combination with one or more compounds of formula I include mevastatin and related compounds (disclosed in U.S. Pat. No. 3,983,140), lovastatin (mevinolin) and related compounds (disclosed in U.S. Pat. No. 4,231,938), pravastatin and related compounds (disclosed, for example, in U.S. Pat. No. 4,346,227), simvastatin and related compounds (disclosed in U.S. Pat. Nos. 4,448,784 and 4,450,171).Other HMG CoA reductase inhibitors that may be used herein include, but are not limited to, fluvastatin (disclosed in U.S. Pat. No. 5,354,772), cerivastatin (disclosed in U.S. Pat. Nos. 5,006,530 and 5,177,080), atorvastatin (disclosed in U.S. Pat. Nos. 4,681,893, 5,273,995, 5,385,929 and 5,686,104), atavastatin (Nissan / Sankyo's nisvastatin), No. 5,011,930), visastatin (Shionogi-Astra / Zeneca (ZD-4522)) (disclosed in U.S. Pat. No. 5,260,440), and related statin compounds (disclosed in U.S. Pat. No. 5,753,675), pyrazole analogs of mevalonolactone derivatives (disclosed in U.S. Pat. No. 4,613,610), indene analogs of mevalonolactone derivatives (disclosed in PCT application WO 86 / 03488), 6-[2-(substituted-pyrrol-1-yl)-alkyl)pyran-2-one and derivatives thereof (disclosed in U.S. Pat. No. 4,647,576), Searle's SC-45355 (3-substituted pentanedioic acid derivatives) dichloroacetate, imidazole analogs of mevalonolactone (disclosed in PCT application WO 86 / 07054), 3-carboxy-2-hydroxy-propane-phosphonic acid derivatives (disclosed in French Patent No. 2,596,393), 2,3-disubstituted pyrrole, furan and thiophene derivatives (disclosed in European Patent Application No. 0221025), naphthyl analogues of mevalonolactone (disclosed in U.S. Pat. No. 4,686,237), octahydronaphthalenes (disclosed, for example, in U.S. Pat. No. 4,499,289), keto analogues of mevinolin (lovastatin) (disclosed in European Patent Application No. 0142146 A2), and quinoline and pyridine derivatives (disclosed in U.S. Pat. Nos. 5,506,219 and 5,691,322).

[0272] Preferred hypolipidemic agents are pravastatin, lovastatin, simvastatin, atorvastatin, fluvastatin, cerivastatin, atavastatin and ZD-4522.

[0273] Additionally, phosphinic acid compounds useful for inhibiting HMG CoA reductase, such as those disclosed in GB 2205837, are suitable for use in combination with the compounds of the present invention.

[0274] Squalene synthetase inhibitors suitable for use herein include, but are not limited to, α-phosphono-sulfonates (disclosed in U.S. Pat. No. 5,712,396), those disclosed in Biller et al, J. Med. Chem., 1988, Vol. 31, No. 10, pp 1869-1871, such as isoprenoid (phosphinyl-methyl) phosphonates, as well as other known squalene synthetase inhibitors (disclosed, for example, in U.S. Pat. Nos. 4,871,721 and 4,924,024 and Biller, SA, Neuenschwander, K., Ponpipom, MM, and Poulter, CD, Current Pharmaceutical Design, 2, 1-40 (1996)).

[0275] Additionally, other squalene synthetase inhibitors suitable for use herein include terpenoid pyrophosphates (disclosed by P. Ortiz de Montellano et al, J. Med. Chem., 1977, 20, 243-249), farnesyl diphosphate analog A and presqualene pyrophosphate (PSQ-PP) analogs (disclosed by Corey and Volante, J. Am. Chem. Soc., 1976, 98, 1291-1293), phosphinyl phosphonates (reported by McClard, RW et al, JACS, 1987, 109, 5544) and cyclopropanes (Capson, TL, PhD dissertation, June, 1987, Dept. Med. Chem. U of Utah, Abstract, Table of Contents, pp 16, 17, 40-43, 48-51, reported by the Summary).

[0276] Fibric acid derivatives which may be used in combination with one or more compounds of formula I include fenofibrate, gemfibrozil, clofibrate, bezafibrate, ciprofibrate, clinofibrate, and the like, probucol and related compounds (disclosed in U.S. Pat. No. 3,674,836) (probucol and gemfibrozil are preferred), bile acid sequestrants such as cholestyramine, colestipol, and DEAE-Sephadex (Secholex®, Policexide®), as well as lipostavir (Rhone-Poulenc), Eisai E-5050 (N-substituted ethanolamine derivative), imanixyl (HOE-402), tetrahydrolipstatin (THL), istigmastanylphos-phorylcholine (SPC, Roche), aminocyclodextrin (Tanabe Seiyoku), Ajinomoto Included are AJ-814 (azulene derivative), melinamide (Sumitomo), Sandoz 58-035, American Cyanamid CL-277,082 and CL-283,546 (disubstituted urea derivatives), nicotinic acid, acipimox, acifuran, neomycin, p-aminosalicylic acid, aspirin, poly(diallylmethylamine) derivatives (disclosed, for example, in U.S. Pat. No. 4,759,923), the quaternary amines poly(diallyldimethylammonium chloride) and ionenes (disclosed, for example, in U.S. Pat. No. 4,027,009), and other known serum cholesterol lowering agents.

[0277] ACAT inhibitors that may be used in combination with one or more compounds of formula I include those described in Drugs of the Future 24, 9-15 (1999), (avasimibe); “The ACAT inhibitor, Cl-1011 is effective in the prevention and regression of aortic fatty streak area in hamsters”, Nicolosi et al, Atherosclerosis (Shannon, Ireland). (1998), 137(1), 77-85; “The pharmacological profile of FCE 27677: a novel ACAT inhibitor with potent hypolipidemic activity mediated by selective suppression of the hepatic secretion of ApoB100-containing lipoprotein”, Ghiselli, Giancarlo, Cardiovasc. Drug Rev. (1998), 16(1), 16-30; “RP 73163: a bioavailable alkylsulfinyl-diphenylimidazole ACAT inhibitor”, Smith, C., et al, Bioorg. Med. Chem. Lett. (1996), 6(1), 47-50; “ACAT inhibitors: physiologic mechanisms for hypolipidemic and anti-atherosclerotic activities in experimental animals”, Krause et al, Editor(s): Ruffolo, Robert R., Jr.; Hollinger, Mannfred A., Inflammation: Mediators Pathways (1995), 173-98, Publisher: CRC, Boca Raton, Fla.; "ACAT inhibitors: potential anti-atherosclerotic agents", Sliskovic et al, Curr. Med. Chem. (1994), 1(3), 204-25; "Inhibitors of acyl-CoA:cholesterol O-acyl transferase (ACAT) as hypocholesterolemic agents. 6. The first water-soluble ACAT inhibitor with lipid-regulating activity. Inhibitors of acyl-CoA:cholesterol acyltransferase (ACAT). 7. Development of a series of substituted N-phenyl-N'-[(1-phenylcyclopentyl)methyl]ureas with enhanced hypocholesterolemic activity", Stout et al, Chemtracts: Org. Chem. (1995), 8(6), 359-62, or TS-962 (Taisho Pharmaceutical Co. Ltd). The hypolipidemic agent may be an upregulator of LD2 receptor activity, such as MD-700 (Taisho Pharmaceutical Co. Ltd) and LY295427 (Eli Lilly).

[0278] Examples of suitable cholesterol absorption inhibitors for use in combination with the compounds of the invention include SCH48461 (Schering-Plough) and those disclosed in Atherosclerosis 115, 45-63 (1995) and J. Med. Chem. 41, 973 (1998).

[0279] Suitable Ileal NaCl for use in combination with the compounds of the present invention + / Examples of bile acid cotransporter inhibitors include the compounds disclosed in Drugs of the Future, 24, 425-430 (1999).

[0280] Lipoxygenase inhibitors that may be used in combination with one or more compounds of Formula I include 15-lipoxygenase (15-LO) inhibitors, such as benzimidazole derivatives (disclosed in WO 97 / 12615), 15-LO inhibitors (disclosed in WO 97 / 12613), isothiazolones (disclosed in WO 96 / 38144), and 15-LO inhibitors (disclosed by Sendobry et al "Attenuation of diet-induced atherosclerosis in rabbits with a highly selective 15-lipoxygenase inhibitor lacking significant antioxidant properties", Brit. J. Pharmacology (1997) 120, 1199-1206, and Cornicelli et al, "15-Lipoxygenase and its Inhibition: A Novel Therapeutic Target for Vascular Disease", Current Pharmaceutical Design, 1999, 5, 11-20).

[0281] Examples of suitable antihypertensive agents for use in combination with the compounds of the invention include beta adrenergic blockers, calcium channel blockers (L-type and T-type; e.g., diltiazem, verapamil, nifedipine, amlodipine, and mibefradil), diuretics (e.g., chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide, bendroflumethiazide, methylchlorothiazide, trichloromethiazide, polythiazide, benzthiazide, ethacrynic acid triclinafen, tricrynafen), chlorthalidone, furosemide, musolimine, bumetanide, triamtrenene, amiloride, spironolactone, renin inhibitors, ACE inhibitors (e.g., captopril, zofenopril, fosinopril, enalapril, ceranopril, cilazopril, delapril, pentopril, quinapril, ramipril, lisinopril), AT-1 receptor antagonists (e.g., losartan, irbesartan, valsartan), ET receptor antagonists (e.g., sitaxsentan, atrsentan, and compounds disclosed in U.S. Pat. Nos. 5,612,359 and 6,043,265), dual ET / AII antagonists (e.g., WO 00 / 01389), neutral endopeptidase (NEP) inhibitors, vasopepsidase inhibitors (dual NEP-ACE inhibitors) (e.g., omapatrilat and gemopatrilat), and nitrates.

[0282] Examples of suitable anti-obesity agents for use in combination with the compounds of the invention include NPY receptor antagonists, NPY-Y2 or NPY-Y4 receptor agonists, MCH antagonists, GHSR antagonists, CRH antagonists, beta 3 adrenergic agonists, lipase inhibitors, serotonin (and dopamine) reuptake inhibitors, thyroid receptor beta drugs, CB-1 antagonists and / or appetite suppressants.

[0283] Beta 3 adrenergic agonists that may optionally be used in combination with the compounds of the invention include AJ9677 (Takeda / Dainippon), L750355 (Merck), or CP331648 (Pfizer) or other known beta 3 agonists (disclosed in U.S. Pat. Nos. 5,541,204, 5,770,615, 5,491,134, 5,776,983, and 5,488,064), with AJ9677, L750,355, and CP331648 being preferred.

[0284] Examples of lipase inhibitors that may optionally be used in combination with the compounds of the present invention include orlistat or ATL-962 (Alizyme), with orlistat being preferred.

[0285] Serotonin (and dopamine) reuptake inhibitors that may optionally be used in combination with the compounds of formula I may be sibutramine, topiramate (Johnson & Johnson) or axokine (Regeneron), with sibutramine and topiramate being preferred.

[0286] Examples of thyroid receptor beta compounds which may optionally be used in combination with the compounds of the present invention include thyroid receptor ligands, such as those disclosed in WO97 / 21993 (U. Cal SF), WO99 / 00353 (KaroBio) and GB98 / 284425 (KaroBio), with the compounds in the KaroBio applications being preferred.

[0287] Examples of CB-1 antagonists that may optionally be used in combination with the compounds of the invention include CB-1 antagonists and rimonabant (SR141716A).

[0288] Examples of NPY-Y2 and NPY-Y4 receptor agonists include PYY(3-36) and pancreatic polypeptide (PP), respectively.

[0289] Appetite suppressants that may optionally be used in combination with the compounds of the present invention include dexamphetamine, phentermine, phenylpropanolamine or mazindol, with dexamphetamine being preferred.

[0290] Examples of suitable antipsychotic agents include clozapine, haloperidol, olanzapine (Zyprexa®), Prozac®, and aripiprazole (Abilify®).

[0291] The aforementioned patents and patent applications are incorporated herein by reference. The above-mentioned other therapeutic agents, when used in combination with the compounds of the present invention, may be used, for example, in the amounts set forth in the Physician's Desk Reference, in the amounts in the above-mentioned patents, or as otherwise determined by one of ordinary skill in the art.

[0292] It will be appreciated by those of ordinary skill in the art that other suitable modifications and adaptations to the compositions and methods described herein will be readily apparent from the description of the invention contained herein in light of the information known to those of ordinary skill in the art, and may be made without departing from the scope of the invention or any embodiment thereof. Having now described the invention in detail, the invention will be more clearly understood by reference to the following examples, which are included herein for illustrative purposes only and are not intended to limit the invention. EXAMPLES

[0293] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0294] Example 1: Peptide design framework The structure-based design and structure-activity analysis of a series of α-helical biased peptidomimetics of the N-terminus of GLP-1 (Tables 1, 2 and 3) provided a framework to inform the optimization of iterative chemical modifications to evolve novel peptide analogs with drug-like properties, including high potency, metabolic stability and improved pharmacokinetics via parenteral or oral drug delivery.

[0295] Aib 2 (Ala replacement), α-MePhe(2,6-F) 6 (Replaces Phe), Bip 10 (Replace Val) and Bip 11 A significant modification of the N-terminal fragment (i.e., His) containing 11 amino acids (replaced Ser) 1 -Ala 2 -Glu 3 -Gly 4 -Thr 5 -Phe 6 -Thr 7 -Ser 8 -Asp 9 -Val 10 -Ser 11 A simplification of GLP-1 to ~) has been previously reported (21-23). ​​To inform the optimization of novel N-terminal GLP-1 fragment analogues, 21 analogues of the general structure H2N-His-Aib-Glu-Gly-Thr-Xaa6-Thr-Ser-Asp-Val-Ser-C(O)NH2 were synthesized and tested for their GLP-1 receptor functional activity (EC 50 , cAMP assay, see below). As shown in Table 1, significantly increased potency was observed for Phe(2-F), Phe(2.6-F), Phe(2,3,4,5,6-F), α-MePhe and α-MePhe(2-F). These data confirm what has been described previously and extend the known structure-activity relationships of numerous other Phe analogs with side chain modifications by various substituents, as well as homologation (to Hph), chiral inversion (to D-Phe), removal of the phenyl ring (to Ala) or substitution (to Bip or Aib). Phe6 and Ala 6 Both were observed to be significantly less potent than their α-Me modified amino acid analogs (i.e., α-MePhe and Aib, respectively). These data provide information about the impact of helix induction by α-methylation that can be achieved within the core N-terminal 11 amino acid sequence of GLP-1.

[0296] [Table 1]

[0297] To obtain information for the optimization of novel GLP-1 analogs, Ala-scanning was performed for the first time on this series of N-terminal GLP-1 fragment analogs (Table 2). The structure-activity relationship of the 10 peptides was characterized by the His 1 , Gly 4 and Thr 7 Substitution of Ala resulted in a >100-fold decrease in potency, whereas substitution of Aib 2 , Glu 3 , Thr 5 , Ser 8 , and Asp 9 It was shown that replacement of Bip by Ala resulted in a >100-fold decrease in potency. 10 and Bip 11 Such data provide information on the impact of simplification of amino acid side chains to methyl groups (Ala), and the efficacy of some amino acids (e.g., Glu 5 , Thr 5 , Ser 8 , and Asp 9 ) provide information that may be amenable to further modifications to adjust their hydrophilic nature (e.g., H-bonding and charge) and helical tendencies (see below; Aib-scanning) to allow for optimization of drug-like properties within Formula I.

[0298] [Table 2]

[0299] To obtain information for the optimization of novel GLP-1 analogs, Aib-scanning was also performed for the first time on this series of N-terminal GLP-1 fragment analogs (Table 3). The structure-activity relationships of the eight peptides were 4 , Thr 7 , Ser 8 Substitution of Aib for Asp9 and His resulted in a >1000-fold decrease in potency, while substitution of Aib for His 1 , Glu 3 , Gly 4 , and Thr 5 We showed that replacement of His1 by Aib resulted in a >200-fold decrease in potency. Indeed, replacement of His1 by Aib was surprisingly potent (only a <30-fold difference). These data support the notion that replacement of His1 by α-methylation or, in some cases, by Aib (e.g., His 1 , Glu 3 or Thr 5 These data further indicate that α-methylation is involved in nucleating and / or maintaining the helical character known from the X-ray structures of GLP-1 and N-terminal fragment analogs (see below), and that incorporation of such α-methylation may allow optimization of drug-like properties within Formula I.

[0300] [Table 3] TIFF2024525236000073.tif45164

[0301] Example 2: GLP-1 peptide agonists incorporating chemical modifications to achieve drug-like properties Exemplary in Tables 4, 5 and 6 are novel N-terminal GLP-1 peptides representative of the scope of Formula 1, illustrating the design and structure-activity characteristics of three series of analogs with N- and / or C-terminal modifications. 1Peptides with N-terminal modifications including extensions of more than 2-1, 2-2, 2-3, 2-4, and 2-5 were designed from computer modeling studies (see below) and were predicted to bind to the GLP-1 receptor (Table 4). These exemplary peptides demonstrated GLP-1 receptor agonist functional potency within 3-fold of the parent peptide analog (1-1). Additionally, the His 1 and Glu 3 Both substitutions as well as His 1 Peptides with extensions of more than 100 fold (e.g., 2-6, 2-7, 2-8, and 2-9) were designed from computer modeling studies (see below) and were predicted to bind to the GLP-1 receptor (Table 1). These exemplary peptides demonstrated GLP-1 receptor agonist functional potency within 100-fold of the parent peptide analog (1-2). Additionally, peptides with substitutions of Aib2 (3-38, 3-39, and 3-40) showed similar potency (e.g., Pro agonist activity) to the parent peptide analog (1-2). 2 ), slightly lower potency (Val 2 ), or significantly greater efficacy (α-MePro 2 ) are shown (Table 4). Additionally, a series of exemplary peptides incorporating combinations of the above N-terminal modifications are listed (Table 4). Taken together, such described N-terminal modifications allow the selection of GLP-1 peptide analogs with superior drug-like properties with respect to agonist potency, metabolic stability, GLP-1 receptor (and GLP-1 receptor family) selectivity (and co-selectivity), and biophysical properties (e.g., helicity, solubility, and hydrophobicity / hydrophilicity).

[0302] [Table 4] TIFF2024525236000075.tif176170

[0303] Specifically, Bip 10 -Bip 11 Peptides incorporating the C-terminal modification of, for example, but not limited to, Hph 10 -Bip 11, Bip 10 -Hph 11 , Bip 10 -α-MeHph 11 , α-MeHph 10 -Bip 11 , Bip 10 -Hph(4-OH) 11 and Bip 10 -NH-(CH2)3-phenyl (see, e.g., 1-3, 1-4, 4-1, 4-2, 4-3, 4-4, 4-4a) were designed as GLP-1 receptor agonists (Table 5). These exemplary peptides exhibited GLP-1 receptor agonist functional potencies ranging from 30-600-fold over the parent peptide (3-1). C-terminal replacement of the carboxamide with a carboxylic acid (4-1) or hydrogen was less potent (approximately 30-fold or >1,000-fold, respectively) relative to their parent peptide analogs (1-4 and 3-1, respectively). Additionally, novel modified Cys 10 or Cys 11 or its α-MeCys analog may be synthetically converted to thioethers within a range of S-aryl, S-heteroaryl, S-heterocyclyl, and S-cycloalkyl groups (e.g., 4-5 to 4-36, Table 5). Additionally, a series of exemplary peptides incorporating combinations of the above N-terminal modifications are listed (Table 5). Taken together, such described N-terminal modifications allow for the selection of GLP-1 peptide analogs with superior drug-like properties with respect to agonist potency, metabolic stability, GLP-1 receptor (and GLP-1 receptor family) selectivity (and co-selectivity), and biophysical properties (e.g., helicity, solubility, and hydrophobicity / hydrophilicity).

[0304] [Table 5] TIFF2024525236000077.tif84170

[0305] In addition, Bip 11Peptides with C-terminal modifications including extensions of more than 10 ... A series of peptide analogs of 1-4 (e.g., 4-35, 4-36, and 4-37) exemplify C-terminal backbone extension via linker (AEEA)2 and conjugation to PK modifiers (C18 diacid or 4-I-phenylpropionic acid) via Dap, as shown in Table 6, and the R for C-terminal modifications incorporating linker and PK modifier conjugates. Y1 , R Y2 and R Y3 The designations (see below) indicate the functional potency of GLP-1 receptor agonists. The two peptides incorporating PK modifiers exhibited functional potencies as GLP-1 receptor agonists ranging from equivalent potency (4-37) to 15-fold less potency (4-36) than the parent peptide (1-4). C-terminal replacement of the carboxamide with a carboxylic acid (4-1) or hydrogen was less potent (approximately 30-fold or >1,000-fold, respectively) relative to their parent peptide analogs (1-4 and 3-1, respectively).

[0306] [Table 6] TIFF2024525236000079.tif219170TIFF2024525236000080.tif22170

[0307] Example 3: Structure-Based Design and Computer Modeling Methodology The general approach to hypothesis-driven peptide design, and many of the computational methods described below, have been described in recent publications (24-27) and have been successfully applied to a variety of protein targets (e.g., IL2R, CXCR4, TLR2, and p53).

[0308] Comparative structural analysis and molecular modeling studies were primarily performed using the cryo-EM structure of the 10 amino acid peptide in complex with the full-length GLP-1R [5nx2] (28), complemented by complexes bound to exendin P5 [6b3j] (29), GLP-1 [5vai] (30) and [6x18] (31), small molecules [6x19 and 6x1a] (31), [6orv] (32), [6xos] (33), [7c2e] (34), and [7lci, 7lcj, 7lck] (35), and the allosteric modulator [6vcb] (36).

[0309] Computational design of novel GLP-1 peptide analogs was accomplished using a hypothesis-driven design approach. This required us to build an initial structural model of a key 11 amino acid peptide located in the GLP-1R active site. This task was accomplished by building a model of the 11 amino acid lead peptide using available X-ray and EM structures as templates. This was followed by constrained conformational optimization. Promising sites for peptide mutations were then identified, and appropriate standard or nonstandard amino acid libraries were defined and constructed. The structures of the novel peptides were then constructed by mutating the peptide ligand from the above complex using a computer program implemented in Python, with the YASARA molecular modeling program (37). The resulting poses were refined using molecular docking with VINA (38), local conformational sampling routines in Python / YASARA (24), or a proprietary Monte Carlo conformational search program written in C++ (Sampler) (24). When constructing analogs with significant structural differences from the reference complex, the initial poses generated by the mutation program were subjected to a short molecular dynamics relaxation step in which the protein backbone was kept fixed. Designed analogs were appropriately rescored. Rescoring was performed by calculating MM and MM / PBSA binding energies. Furthermore, models were visually inspected to ensure that they were not biased by artifacts of the computational method.

[0310] To enhance the structural understanding of GLP-1R binding to the peptide and to inform future design rounds, the models were qualitatively and quantitatively analyzed against the experimentally measured cAMP EC50 results. The structure-activity data were visually analyzed to identify patterns. The structure-activity data were also subjected to quantitative structure-activity relationship (QSAR) analysis using ligand-based and receptor-ligand-based approaches. One ligand-based approach used molecular field analysis using Cresset Forge (39). Another ligand-based approach used molecular field analysis using Cresset Forge (https: / / www.cresset-group.com / sofware / forge / ). For the receptor-ligand QSAR analysis, the receptor-ligand interface descriptors were calculated using a proprietary YASARA script.

[0311] An initial QSAR analysis was performed on selected structure-activity relationship data. In particular, a GLP-1 peptide / GLP-1R all-atom structural model based on the 5nx2 crystal structure was used to calculate various receptor-based and ligand-based descriptors for 28 GLP-1 peptide / GLP-1R structural variants. Multiple regression analysis was performed using pEC50 measurements as dependent variables. The GLP-1 peptide variants covered Ala-scan, Aib-scan, truncated analogs, and N-terminal extension analogs (see above). The final descriptor-based regression equation included three molecular descriptors: pEC50 = -0.012YSCORE + -0.91Ion-Ion Energy + -0.19Backbone Torsions + Constant where all descriptors were calculated using the YASARA molecular modeling software package, YSCORE refers to the binding energy calculated using the NOVA2 force field, Ion-Ion refers to the electrostatic energy between ion-ion interface contacts, and BackboneTorsions refers to the total number of phi / psi torsions in the peptide (40). The overall model was found to be statistically significant. Individual terms were also found to contribute statistically significantly to the pEC50 estimates. The resulting best-fit line is shown in Figure 1. The results (R 2 = 0.65) is encouraging, especially considering that the QSAR equation is based on only three a priori physically and structurally reasonable descriptors that contribute statistically significantly to the pEC50 estimation. Such studies will provide a focus for future training and testing of QSAR models.

[0312] Example 4: Peptide synthesis The polypeptides of the invention were prepared using the following methods for coupling the appropriate amino acids. Deprotection, cleavage and purification methods are also described.

[0313] Peptide synthesis, purification, and analysis: Solid phase peptide synthesis was accomplished by standard methods. Typically, Amphispheres 40 RAM, 75-150 μM resin (Agilent Technologies) was used to generate peptides as C-terminal carboxamides. The amino acid coupling protocol using HCTU generally included four steps: (a) 1st coupling - 5 eq amino acid (0.34 M), 10 eq DIEA (2 M), 5 eq HCTU (0.5 M), 5 eq 6-Cl-HOBt (0.5 M), 30 min; (b) 2nd coupling - 5 eq amino acid (0.34 M), 10 eq DIEA (2 M), 5 eq HCTU (0.5 M), 5 eq 6-Cl-HOBt (0.5 M), 90 min; (c) 1 DMF wash between couplings; and (d) 9 DMF washes after the 2nd coupling. The amino acid coupling protocol using HATU generally involved two steps: (a) single coupling - 2 eq amino acid (0.1 M), 4 eq DIEA (2 M), 2 eq HATU (0.5 M), 5 eq HOAt (0.5 M), 240 min; and (b) 9 DMF washes after coupling. The amino acid coupling protocol using PyOxim and HATU generally included 4 steps: (a) 1st coupling - 5 eq amino acid (0.34M), 10 eq DIEA (2M), 5 eq PyOxim (0.5M), 120 min; (b) 2nd coupling - 5 eq amino acid (0.34M), 10 eq DIEA (2M), 5 eq HCTU (0.5M), 5 eq HOAt (0.5M), 120 min; (c) 1 DMF wash between couplings; (d) 9 DMF washes after 2nd coupling. The Fmoc deprotection protocol generally included 3 steps: (a) 20% piperidine in DMF, 10 min; (b) 20% piperidine in DMF, 15 min; and (c) 8 DMF washes.Cleavage of the amino acid side chain protecting groups and peptide from the resin was typically accomplished by the following five steps: (a) cleavage cocktail of 87.5% TFA, 2.5% anisole, 5% water, 5% triisopropylsilane, 3-4 h, 10 mL per gram of resin; (b) modified procedure for sulfur-containing amino acids: 85% Tfa, 2.5% 3,6-dioxa-1,8-octanedithiol, 2.5% anisole, 5% water, 5% triisopropylsilane, 3-4 h, 10 mL per gram of resin; (c) evaporating the TFA; (d) precipitation with cold diethyl ether (minimum 10:1, ether:cleavage cocktail), centrifugation at 3000 rpm for 5 min, then decanting the ether (this was repeated three times); and e) the peptide powder / pellet was then dried overnight. Purification by reversed-phase HPLC was achieved in four steps: (a) resolving the peptide; (b) chromatography using a Biotage Selekt instrument and a Biotage Sfar Bio C18 D (Duo, 300 Å, 20 μm); (c) pooling, freezing and lyophilizing the desired fractions; and (d) adding 50% acetonitrile / water to the dried peptide, which was then refrozen and lyophilized again. Analysis of purified peptides was accomplished in three steps: (a) peptide samples were dissolved and analyzed using an Agilent Infinity II LC / MS; (b) analysis for purity (absorbance detection at 214 and 280 nm) and retention time using an Agilent Zorbax 300SB-C18 (5 μm, 2.1 × 150 mm) and a two solvent system consisting of A (0.1% TFA in water) and B (0.1% Tfa in acetonitrile) (gradient of 5-65% B over 20 min at 40 °C); and MS analysis using an Agilent Infinity Lab MSD, positive polarity (mass detection range is 100-1500).

[0314] Amino acids illustrating S-perfluoroarylation (e.g., Cys-S-biphenyl [F9]): Crude or purified Cys-containing peptides are dissolved in DMF containing an excess of an electrophile (e.g., decafluorobiphenyl) and a base, such as N,N-diisopropylethylamine (DIEA) or Tris, to obtain a concentration of 1 mM. Once the reaction is complete, it is quenched with a thiol and purified by high performance liquid chromatography (HPLC).

[0315] New Amino Acids Exemplifying the Azide / Alkyne Huisgen Cycloaddition ("Click"): Procedure for peptide conjugation by azide / alkyne Huisgen cycloaddition: Peptide azide and lipid-alkyne are added to a glass scintillator sealed with a septum cap along with copper bromide. The reaction mixture is purged with nitrogen for 5 minutes to ensure the removal of oxygen, and then approximately 1 mL of degassed DMF is added. The reaction mixture is vortexed. The reaction is allowed to continue for 2 hours and then purified by RP-HPLC.

[0316] Resolute Bio's SOP-0002 specifies the following synthetic steps for the on-resin cycloaddition reaction between a protected, fully assembled peptide-resin position #10 or position #11 Phe(4-azide) and an alkyne-fatty acid (e.g., 17-octadecynoic acid) using CuAAC reagent on a 0.1 mmol scale, prior to purification by RP-HPLC: (1) swell the resin with DMF; (2) wash the resin three times with 20% 2,6-lutidine in DMF; (3) add 1.5 equivalents of the alkyne reagent (or azide reagent if coupling to an alkyne); (4) add 49.5 mg of sodium ascorbate; (5) add 45 μL of DIEA; (6) add 47.5 mg of CuI; and (7) stir overnight at ambient temperature. Purification of RXL-4042-2 and RXL-4043-2 was achieved using an Agilent 1290 Prep System equipped with an Agilent 1260 Multi-Wavelength Detector. The fraction trigger was set at a wavelength of 214 nm. The reverse phase chromatography column was an Agilent Prep, 100A, 5 μm, C18, 50×21.2 mm. The structure of 17-octadecynoic acid is shown below:

[0317] [ka]

[0318] Novel PK modified peptides illustrating albumin binding functional group conjugation: Procedure for peptide conjugation with a PK modifier, e.g., a serum albumin binding group: The N-terminus of the peptide is conjugated to two primary amine positions (e.g., His 1 Against R Y2 ~R Y12The linkers, represented in Figure 1, require acetylation or Boc protecting groups to prevent amide formation at Dap [diaminopropionic acid] or Glu amino groups). Note that such PK modifiers can be conjugated to the L- or D-enantiomers of Dap or Glu, as well as other linker-related amino acids, such as Lys, Orn (ornithine), or Dab (diaminobutyric acid). In such cases, the primary amine moiety is protected with Mtt (methyltrityl) or Mmt (methoxytrityl) by the following protocol: (a) wash the resin three times with 2% trifluoroacetic acid, 2% triisopropylsilane, and 96% DCM; (b) shake twice for 30 minutes with 2% trifluoroacetic acid, 2% triisopropylsilane, and 96% DCM; wash three times with DCM (dichloromethane). After removal of Mtt or Mmt, wash the resin three times with 2% DIEA (diisopropylethylamine). Wash 3 times with DMF (N,N-dimethylformamide). Wash 3 times with NMP (N-methyl-2-pyrrolidinone). The amine can then be generated by direct conjugation with a PK modifier (e.g., C18 fatty acid or aryl halide) or linker group (e.g., AEEA), typically using 2 equivalents (PK modifier or AEEA / amino acid linker) with 2 equivalents of PyOxim and 4 equivalents of DIEA. To test for completeness of coupling, microcleave of the peptide-resin may be performed to determine if a further recoupling step is required.

[0319] Example 5: In vitro pharmacological screening method In this assay, human embryonic kidney cells (HEK) co-expressing the hGLP1 receptor and a CRE-luciferase construct were used to determine the potency of agonists. Cells were briefly thawed at 37°C, transferred to a sterile tube, and resuspended in complete medium at 37°C. Cells were centrifuged at 1000 rpm for 5 min to collect the cells; cells were resuspended in assay buffer consisting of DPBS (GIBCO) with 500 μM of the phosphodiesterase inhibitor IBMX. The assay medium may contain serum albumin (2%) to test for albumin affinity, or no albumin as specified in the particular protocol. The optimal cell density was determined to be 1000 cells / well; cells were added to wells in a 384-well plate containing the appropriate pre-prepared dilutions of the compound (test peptide or reference compound Exendin-4), sealed, and incubated with CO2 for 30 min. Test peptide solutions were diluted from a 10 mM stock solution; for most peptides, an initial run was performed in duplicate from a maximum concentration of 1.0 μM, and 11 concentrations were tested for each peptide using a 1:3 serial dilution from this maximum concentration. With peptides found to be particularly potent agonists, subsequent assays were run using a maximum concentration of 1.0 nM (11 concentrations, 1:3 dilution from 1 nM). The agonist assay was a homogeneous time-resolved fluorescence (HTRF) assay (Cisbio).

[0320] After incubating the cells with the test or reference peptides for 30 min, 5 μL of the cAMP acceptor cAMP-d2, previously prepared as a working solution from frozen stock (1:20 dilution), was added to each well of the assay plate along with 5 μL of anti-cAMP antibody-cryptate working solution (1:20 dilution from frozen stock). The wells were incubated for 1 h at room temperature and then the fluorescence was read at 665 and 615 nm using an Envision reader equipped with a TRF laser. The data were stored and analyzed using Prism software (GraphPad). A concentration-response analysis was performed using a four-parameter logistic fit of the resulting data to obtain EC values ​​for each test compound and the reference compound. 50 The value was obtained.

[0321] Incorporation by Reference All U.S. patents and U.S. patent application publications cited herein are hereby incorporated by reference.

[0322] Equivalent Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein which equivalents are intended to be encompassed by the following claims.

[0323] References cited TIFF2024525236000082.tif28160TIFF2024525236000083.tif223163TIFF2024525236000084.tif223163TIFF2024525236000085.tif200167

Claims

1. The following array: R XN -X aa 1 -X aa 2 -X aa 3 -X aa 4 -X aa 5 -X aa 6 -X aa 7 -X aa 8 -X aa 9 -X aa 10 -X aa 11 -R YC [wherein, R XN is H and -N(Rx) 2 (wherein each Rx is independently H, optionally substituted alkyl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted formyl, optionally substituted acetyl, optionally substituted alkanoyl, optionally substituted -C(O)-alkyloxy, optionally substituted -C(O)-aryloxy, optionally substituted -C(O)-aralkyloxy, optionally substituted -C(O)-heterocyclyloxy, optionally substituted -C(O)-heteroarylalkyloxy, optionally substituted -C(O)NH-alkyl, optionally substituted -C(O)NH-aryl, optionally substituted -C(O)NH-arylalkyl, optionally substituted -SO 2 -heterocyclyl, optionally substituted -SO 2 -alkyl, optionally substituted -SO 2 -aryl, optionally substituted -SO 2 -arylalkyl, optionally substituted -SO 2 -heteroarylalkyl, optionally substituted -SO 2-selected from the group consisting of heteroaryl and optionally substituted ureido; or the first Rx is hydrogen and the second Rx is an amino acid residue X aa 0 which is an N-terminal group of X selected from the group consisting of aa 1 ; X aa 0 is an optionally substituted amino acid residue selected from the group consisting of Gly, Pro, Arg, Glu, His, Phe, Trp, and Aib; X aa 1 is an optionally substituted amino acid residue containing an amino acid side chain containing an alkyl, aryl or heteroaryl group; X aa 2 is an optionally substituted amino acid residue selected from the group consisting of Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; X aa 3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 4 is an amino acid residue selected from the group consisting of Gly, Ala, Aib, and β-Ala; X aa 5 is an optionally substituted amino acid selected from the group consisting of Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; X aa 6 is an optionally substituted amino acid residue that is disubstituted at the α-carbon, and one of the α-carbon substituents is an optionally substituted aryl or optionally substituted heteroaryl; X aa 7is an optionally substituted amino acid residue containing an amino acid side chain containing a hydroxyl group; X aa 8 is an optionally substituted amino acid residue selected from the group consisting of Ser, His, and Asn; X aa 9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 10 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide, optionally substituted aryl, optionally substituted heteroaryl, or any combination thereof; X aa 11 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide group, optionally substituted aryl, optionally substituted heteroaryl, or any combination thereof; and R YC is a structure - C(O)N(R Y ) 2 (wherein each R Y is independently hydrogen or a PK modifying group, where the PK modifying group improves the pharmacokinetic profile of the polypeptide) and is the C - terminal group of Xaa 11 )] A polypeptide represented by.

2. X aa 1 is an optionally substituted amino acid residue containing an amino acid side chain containing (C 1 ~C 4 )alkyl, imidazole, or phenyl, and optionally, Xaa1 is an optionally substituted amino acid residue selected from Leu, His, and Tyr. The polypeptide according to claim 1. **Claim 3**: The polypeptide according to claim 1, wherein Xaa2 is selected from the group consisting of Aib, Pro, α-methyl-Pro, and Val, or Xaa2 is substituted with at least one halogen or alkyl group. **Claim 4** X aa 3 is an amino acid residue selected from the group consisting of Asp, Glu, and cysteic acid, or Xaa3 is substituted with at least one halogen or alkyl group, the polypeptide according to claim 1. **Claim 5** X aa 4 is an amino acid residue selected from the group consisting of Gly and Ala, the polypeptide according to claim 1. **Claim 6** X aa 5 is unsubstituted or substituted Thr, or Xaa5 is substituted with at least one halogen or alkyl group, the polypeptide according to claim 1. **Claim 7** X aa 6 is **Chemical Formula 1** [wherein, X 6a is an alkyl group; and X 6b is a substituted arylalkyl group] is an optionally substituted amino acid residue represented by, or Xaa6 is an optionally substituted amino acid residue selected from α-MePhe, α-MePhe(2-F), and α-MePhe(2,6-DiF), the polypeptide according to claim 1. **Claim 8** X aa 7 is an optionally substituted amino acid residue selected from the group consisting of Thr, α-MeThr, Ser, and α-MeSer, or Xaa7 is substituted with at least one halogen or alkyl group, the polypeptide according to claim 1.

9. X aa 8 is Ser which may be substituted, or Xaa8 is substituted with at least one halogen or alkyl group, the polypeptide according to claim 1.

10. X aa 9 is an amino acid residue selected from the group consisting of Asp, Glu and cysteic acid, or Xaa9 is substituted with at least one halogen or alkyl group, the polypeptide according to claim 1.

11. X aa 10 is 【Chemical Formula 2】 [wherein, X 1 is -N- or -CR10b-; X 2 is -N- or -CR10c-; X 3 is -N- or -CR10d-; X 4 is -N- or -CR10e-; X 5 is -N- or -CR10f-; Z 1 is selected from the group consisting of a bond, -S- and -SO2-; R 10a is H or alkyl; and R 10b 、R 10c 、R 10d 、R 10e 、and R 10f are each independently selected from the group consisting of H, halogen, and alkyl] represented by or Xaa10 is 【Chemical Formula 3】 [wherein, X1 is -N- or -CR10b-; X2 is -N- or -CR10c-; X4 is -N- or -CR10e-; X5 is -N- or -CR10f-; X6 is -N- or -CR10g-; X7 is -N- or -CR10h-; X8 is -N- or -CR10i-; X9 is -N- or -CR10j-; X10 is -N- or -R10k; Z1 is selected from the group consisting of a bond, -S-, and -SO2-; Z2 is selected from the group consisting of a bond, -S-, and -SO2-; R10a is H and alkyl; and R10b, R10c, R10e, R10f, R10g, R10h, R10i, R10j, and R10k are each independently selected from the group consisting of H, halogen, and alkyl; represented by; or Xaa10 is [Chemical Formula 4] [wherein, X1 is -N- or -CR10b-; X2 is -N- or -CR10c-; X4 is -N- or -CR10e-; X5 is -N- or -CR10f-; Z1 is selected from the group consisting of a bond, -CH2-, -S-, -O-, -NH-, -SO2-, -SO2-NH-, -NH-SO2-, -NHC(O)-, and -C(O)NH-; R10a is H or alkyl; R 10d is -L 1 -L 2 -L 3 -R 10d '; R 10d'is -NH 2, (C 1 -C 20 alkyl)-CO 2 H or optionally substituted (C 1 -C 6 alkyl)-aryl; L 1 is a bond or a linker; L 2 is a linker containing an ether moiety; L 3 is a bond or a linker containing an amino acid moiety; and R 10b, R 10c, R 10d, R 10e, and R 10f are each independently selected from the group consisting of H, halogen, and alkyl] represented by, or X aa 10 is [Chemical Formula 5] [wherein, X 1 is -N- or -CR 10b -; X 2 is -N- or -CR 10c -; X 4 is -N- or -CR 10e -; X 5 is -N- or -CR 10f -; X 6 is -N- or -CR 10g -; X 7 is -N- or -CR 10h -; X 9 is -N- or -CR 10j -; X 10 is N or -CR 10k -; Z 1 is selected from the group consisting of a bond, -CH 2 -, -NH-, -S-, -SO 2 -, -O-, -SO 2 -NH-, -NH-SO 2 -, -NHC(O)-, and -C(O)NH-; Z 2 is selected from the group consisting of a bond, -NH-, -S-, -SO 2 -, -O-, -SO 2 -NH-, -NH-SO 2 -, -NHC(O)-, and -C(O)NH-; R 10a is H or alkyl; R 10i is -L 1 -L 2 -L 3 -R 10i '; R 10i' is -NH 2, (C 1 -C 20 alkyl)-CO 2 H or optionally substituted (C 1 -C 6 alkyl)-aryl; L 1 is a bond or a linker; L 2 is a linker containing an ether moiety; L 3 is a bond or a linker containing an amino acid moiety; and R 10b, R 10c, R 10e, R 10f, R 10g, R 10h, R 10j, and R 10k are each independently selected from the group consisting of H, halogen, and alkyl] represented by, or X aa 10 is [Chemical Formula 6] [wherein, X 1 is -N- or -CR 10b -; X 2 is -N- or -CR 10c -; X 4 is -N- or -CR 10e -; X 5 is -N- or -CR 10f -; R 10a is H or alkyl; R 10b, R 10c, R 10e, and R 10f are each independently selected from the group consisting of H, halogen, and alkyl; and Z 3 is a substituted heteroaryl] The polypeptide according to claim 1, represented by.

12. X aa 11 is [Chemical Formula 7] [wherein, X 1 is -N- or -CR 10b -; X 2is -N- or -CR 10c -; X 3 is -N- or -CR 10d -; X 4 is -N- or -CR 10e -; X 5 is -N- or -CR 10f -; Z 1 is selected from the group consisting of a bond, -S-, and -SO2-; R 10a is H or alkyl; and R 10b 、R 10c 、R 10d 、R 10e 、and R 10f are each independently selected from the group consisting of H, halogen, and alkyl] represented by, or Xaa11 is [Chemical Formula 8 [wherein, X1 is -N- or -CR10b-; X2 is -N- or -CR10c-; X4 is -N- or -CR10e-; X5 is -N- or -CR10f-; X6 is -N- or -CR10g-; X7 is -N- or -CR10h-; X8 is -N- or -CR10i-; X9 is -N- or -CR10j-; X10 is -N- or -CR10k-; Z1 is selected from the group consisting of a bond, -S-, and -SO2-; Z2 is selected from the group consisting of a bond, -S-, and -SO2-; R 10a is H or alkyl; and R 10b, R 10c, R 10e, R 10f, R 10g, R 10h, R 10i, R 10j, and R 10k are each independently selected from the group consisting of H, halogen, and alkyl] represented by, or X aa 11 is [Chemical Formula 9] [wherein, X 1 is -N- or -CR 10b -; X 2 is -N- or -CR 10c -; X 4 is -N- or -CR 10e -; X 5 is -N- or -CR 10f -; Z 1 is selected from the group consisting of a bond, -CH 2 -, -S-, -O-, -NH-, -SO 2 -, -SO 2 -NH-, -NH-SO 2 -, -NHC(O)-, and -C(O)NH-; R 10a is H or alkyl; R 10d is -L 1 -L 2 -L 3 -R 10d '; R 10d'is -NH 2, (C 1 -C 20 alkyl)-CO 2 H or optionally substituted (C 1 -C 6 alkyl)-aryl; L 1 is a bond or a linker; L 2 is a linker containing an ether moiety; L 3 is a bond or a linker containing an amino acid moiety; and R 10b, R 10c, R 10d, R 10e, and R 10f are each independently selected from the group consisting of H, halogen, and alkyl] represented by, or X aa 11 is [Chemical Formula 10] [wherein, X1 is -N- or -CR10b-; X2 is -N- or -CR10c-; X4 is -N- or -CR10e-; X5 is -N- or -CR10f-; X6 is -N- or -CR10g-; X7 is -N- or -CR10h-; X9 is -N- or -CR10j-; X10 is -N- or -CR10k-; Z1 is selected from the group consisting of a bond, -CH2-, -NH-, -S-, -SO2-, -O-, -SO2-NH-, -NH-SO2-, -NHC(O)-, and -C(O)NH-; Z2 is selected from the group consisting of a bond, -NH-, -S-, -SO2-, -O-, -SO2-NH-, -NH-SO2-, -NHC(O)-, and -C(O)NH-; R10a is H or alkyl; R10i is -L1-L2-L3-R10i'; R10i' is -NH2, (C1-C20 alkyl)-CO2H or optionally substituted (C1-C6 alkyl)-aryl; L1 is a bond or a linker; L2 is a linker containing an ether moiety; L3 is a bond or a linker containing an amino acid moiety; and R10b, R10c, R10e, R10f, R10g, R10h, R10j, and R10k are each independently selected from the group consisting of H, halogen, and alkyl] represented by, or Xaa11 is [Chemical Formula 11] [wherein, X1 is -N- or -CR10b-; X2 is -N- or -CR10c-; X4 is -N- or -CR10e-; X5 is -N- or -CR10f-; R10a is H or alkyl; R10b, R10c, R10e, and R10f are each independently selected from the group consisting of H, halogen, and alkyl; and Z3 is substituted heteroaryl] The polypeptide according to claim 1, represented by

13. R 10d is 【Chemical Formula 12】 【Chemical Formula】 selected from the group consisting of, or R10i is 【Chemical Formula 13】 【Chemical Formula】 selected from the group consisting of, the polypeptide according to claim 12.

14. X aa 10 is Phe, Tyr, Trp, homophenylalanine (Hph), homotyrosine (Hty), Bip, α-MeBip, 4-phenyl-3-pyridylalanine, 4-phenyl-4-pyridylalanine, α-MeHph, α-MeTyr, α-MeHty, Tyr(O-phenyl), Phe(4-S-phenyl), Phe(4-SO 2 -NH-phenyl), Phe(4-CO-NH-phenyl), Cys(S-phenyl), Cys(S-phenyl[2,3,4,5,6-F 5 )), Cys(S-phenyl[2,3,4,5-F 4 -4-phenyl[2',3',4',5',6'-F 5)), Cys(S-phenyl[2,3,4,5-F 4 -4-SO 2 -phenyl[2',3',4',5',6'-F 5 )), Cy(SO 2 -NH-phenyl), Dap(3-C[=O]-phenyl), Dap(3-[C=O]-pyridyl), Asp(3-NH-phenyl), and Asp(3-NH-pyridyl), or Xaa11 is Phe, Tyr, Trp, homophenylalanine (Hph), homotyrosine (Hty), Bip, α-MeBip, 4-phenyl-3-pyridylalanine, 4-phenyl-4-pyridylalanine, α-MeHph, α-MeTyr, α-MeHty, Tyr(O-phenyl), Phe(4-S-phenyl), Phe(4-SO2-NH-phenyl), Phe(4-CO-NH-phenyl), Cys(S-phenyl), Cys(S-phenyl[2,3,4,5,6-F5]), Cys(S-phenyl[2,3,4,5-F4]-4-phenyl[2',3',4',5',6'-F5]), Cys(S-phenyl[2,3,4,5-F4]-4-SO2-phenyl[2',3',4',5',6'-F5]), Cy(SO2-NH-phenyl), Dap(3-C[=O]-phenyl), Dap(3-[C=O]-pyridyl), Asp(3-NH-phenyl), and Asp(3-NH-pyridyl), and is a optionally substituted amino acid residue selected from the group consisting of, the polypeptide according to claim 1.

15. R YC is -C(O)N(R Y ) 2 [wherein each R Y is H], or RYC is -C(O)N(RY)2, where one occurrence of RY is hydrogen and the other occurrence of RY is -L4-L5-L6-L7-RY'; RY' is (C1-C20 alkyl)-CO2H or optionally substituted (C1-C6 alkyl)-aryl; L4 is a linker containing a bond or an amino acid moiety, and optionally, L4 is 【Chemical Formula 14】 and m is 1, 2, 3, 4, 5 or 6, L5 is a linker containing a bond or an amino acid moiety, and optionally, L5 is 【Chemical Formula 15】 or 【Chemical Formula 16】 and l is 1, 2, 3, 4, 5 or 6, and n is 1, 2, 3, 4, 5 or 6, L6 is a linker containing a bond or an ether moiety, and optionally, L6 is 【Chemical Formula 17】 and L7 is a linker containing an amino acid moiety, The polypeptide according to claim 1.

16. R YC is -C(O)NHR Y and NHR Y is 【Chemical Formula 18】 【Chemical Formula】 【Chemical Formula】 【Chemical Formula】 【Chemical Formula】 The polypeptide according to claim 1, selected from the group consisting of

17. as follows: 【Chemical Formula 19】 【Chemical Formula】 [wherein, Each n is independently 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; Each R 2A is independently -H or alkyl; and Each R 10A is independently -H or alkyl. The polypeptide according to claim 1, represented by

18. as follows: 【Chemical Formula 20】 [wherein, Each m is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; Each R 2A is independently -H or alkyl; Each R 10A is independently -H or alkyl; Each R 10Z is independently halogen, and optionally, R 10Z is I; and Each R 11Z is independently halogen, and optionally, R 11Z is I. The polypeptide according to claim 1, represented by

19. as follows: 【Chemical Formula 21】 【Chemical Formula】 【Chemical Formula】 【Chemical Formula】 The polypeptide according to claim 1, represented by

20. A pharmaceutical composition comprising the polypeptide according to any one of claims 1 to 19 and a pharmaceutically acceptable excipient.

21. The following sequence: RXN-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-RYC [wherein, RXN is selected from the group consisting of H and -N(Rx)2 (wherein each Rx is independently H, optionally substituted alkyl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted formyl, optionally substituted acetyl, optionally substituted alkanoyl, optionally substituted -C(O)-alkyloxy, optionally substituted -C(O)-aryloxy, optionally substituted -C(O)-aralkyloxy, optionally substituted -C(O)-heterocyclyloxy, optionally substituted -C(O)-heteroarylalkyloxy, optionally substituted -C(O)NH-alkyl, optionally substituted -C(O)NH-aryl, optionally substituted -C(O)NH-arylalkyl, optionally substituted -SO2-heterocyclyl, optionally substituted -SO2-alkyl, optionally substituted -SO2-aryl, optionally substituted -SO2-arylalkyl, optionally substituted -SO2-heteroarylalkyl, optionally substituted -SO2-heteroaryl, and optionally substituted ureido); or the first Rx is hydrogen and the second Rx is the amino acid residue Xaa0); and is an N-terminal group of Xaa1 selected from the group consisting of Xaa0 is an optionally substituted amino acid residue selected from the group consisting of Gly, Pro, Arg, Glu, His, Phe, Trp, and Aib; Xaa1 is an optionally substituted amino acid residue containing an amino acid side chain containing an alkyl, aryl or heteroaryl group; Xaa2 is an optionally substituted amino acid residue selected from the group consisting of Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; Xaa3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; Xaa4 is an amino acid residue selected from the group consisting of Gly, Ala, Aib, and β-Ala; Xaa5 is an optionally substituted amino acid selected from the group consisting of Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; Xaa6 is an optionally substituted amino acid residue disubstituted at the α-carbon, and one of the α-carbon substituents is an optionally substituted aryl or an optionally substituted heteroaryl; Xaa7 is an optionally substituted amino acid residue containing an amino acid side chain containing a hydroxyl group; Xaa8 is an optionally substituted amino acid residue selected from the group consisting of Ser, His, and Asn; Xaa9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; Xaa10 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; Xaa11 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide group, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; and RYC is the C-terminal group of Xaa11 having the structure -C(O)N(RY)2 (wherein each RY is independently hydrogen or a PK-modifying group, and the PK-modifying group improves the pharmacokinetic profile of the polypeptide). A composition for use in activating a glucagon-like peptide-1 (GLP-1) receptor, comprising a polypeptide represented by The activation comprises contacting the GLP-1 receptor with the polypeptide. **Claim 22** The following sequence: RXN-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-RYC [wherein RXN is selected from the group consisting of H and -N(Rx)2 (wherein each Rx is independently H, optionally substituted alkyl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted formyl, optionally substituted acetyl, optionally substituted alkanoyl, optionally substituted -C(O)-alkyloxy, optionally substituted -C(O)-aryloxy, optionally substituted -C(O)-aralkyloxy, optionally substituted -C(O)-heterocyclyloxy, optionally substituted -C(O)-heteroarylalkyloxy, optionally substituted -C(O)NH-alkyl, optionally substituted -C(O)NH-aryl, optionally substituted -C(O)NH-aralkyl, optionally substituted -SO2-heterocyclyl, optionally substituted -SO2-alkyl, optionally substituted -SO2-aryl, optionally substituted -SO2-aralkyl, optionally substituted -SO2-heteroarylalkyl, optionally substituted -SO2-heteroaryl, and optionally substituted ureido); or the first Rx is hydrogen and the second Rx is an amino acid residue Xaa0, and is the N-terminal group of Xaa1 selected from the group consisting of Xaa0 is an optionally substituted amino acid residue selected from the group consisting of Gly, Pro, Arg, Glu, His, Phe, Trp, and Aib; Xaa1 is an optionally substituted amino acid residue containing an amino acid side chain containing an alkyl, aryl or heteroaryl group; Xaa2 is an optionally substituted amino acid residue selected from the group consisting of Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; Xaa3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; Xaa4 is an amino acid residue selected from the group consisting of Gly, Ala, Aib, and β-Ala; Xaa5 is an optionally substituted amino acid selected from the group consisting of Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; Xaa6 is an optionally substituted amino acid residue that is disubstituted at the α-carbon, and one of the α-carbon substituents is an optionally substituted aryl or an optionally substituted heteroaryl; Xaa7 is an optionally substituted amino acid residue containing an amino acid side chain containing a hydroxyl group; Xaa8 is an optionally substituted amino acid residue selected from the group consisting of Ser, His, and Asn; Xaa9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; Xaa10 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; Xaa 11 is an optionally substituted amino acid residue comprising an amino acid side chain comprising a sulfide group, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; and RYC is a C-terminal group of Xaa 11 having the structure -C(O)N(RY)2 (wherein each RY is independently hydrogen or a PK modifying group, where the PK modifying group improves the pharmacokinetic profile of the polypeptide). A composition for use in the treatment or prevention of diabetes, comprising a polypeptide represented by: The treatment or prevention comprises administering the polypeptide to a subject in need thereof, Optionally, the diabetes is type II diabetes, Optionally, the administering step is by oral administration or parenteral administration, of the composition. **Claim 23** The following sequence: RXn-Xaa 1-Xaa 2-Xaa 3-Xaa 4-Xaa 5-Xaa 6-Xaa 7-Xaa 8-Xaa 9-Xaa 10-Xaa 11-RYC [wherein, RXN is the N-terminal group of Xaa1 selected from the group consisting of H and -N(Rx)2 (where each Rx is independently H, optionally substituted alkyl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted formyl, optionally substituted acetyl, optionally substituted alkanoyl, optionally substituted -C(O)-alkyloxy, optionally substituted -C(O)-aryloxy, optionally substituted -C(O)-aralkyloxy, optionally substituted -C(O)-heterocyclyloxy, optionally substituted -C(O)-heteroarylalkyloxy, optionally substituted -C(O)NH-alkyl, optionally substituted -C(O)NH-aryl, optionally substituted -C(O)NH-arylalkyl, optionally substituted -SO2-heterocyclyl, optionally substituted -SO2-alkyl, optionally substituted -SO2-aryl, optionally substituted -SO2-arylalkyl, optionally substituted -SO2-heteroarylalkyl, optionally substituted -SO2-heteroaryl, and optionally substituted ureido); or the first Rx is hydrogen and the second Rx is the amino acid residue Xaa0); Xaa0 is an optionally substituted amino acid residue selected from the group consisting of Gly, Pro, Arg, Glu, His, Phe, Trp, and Aib; Xaa1 is an optionally substituted amino acid residue containing an amino acid side chain containing an alkyl, aryl or heteroaryl group; Xaa2 is an optionally substituted amino acid residue selected from the group consisting of Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; Xaa3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; Xaa 4 is an amino acid residue selected from the group consisting of Gly, Ala, Aib, and β-Ala; Xaa 5 is an optionally substituted amino acid selected from the group consisting of Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; Xaa 6 is an optionally substituted amino acid residue that is disubstituted at the α-carbon, and one of the α-carbon substituents is an optionally substituted aryl or an optionally substituted heteroaryl; Xaa 7 is an optionally substituted amino acid residue containing an amino acid side chain containing a hydroxyl group; Xaa 8 is an optionally substituted amino acid residue selected from the group consisting of Ser, His, and Asn; Xaa 9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; Xaa 10 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; Xaa 11 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide group, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; and RYC is a C-terminal group of Xaa 11 having the structure -C(O)N(RY)2 (wherein each RY is independently hydrogen or a PK modifying group, and the PK modifying group improves the pharmacokinetic profile of the polypeptide). A composition for use in the treatment or prevention of obesity, comprising a polypeptide represented by wherein the treatment or prevention comprises administering the polypeptide to a subject in need thereof, and optionally, the administering step is by oral or parenteral administration of the composition.

24. The following sequence: RXN-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-RYC [wherein, RXN is selected from the group consisting of H and -N(Rx)2 (wherein each Rx is independently H, optionally substituted alkyl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted formyl, optionally substituted acetyl, optionally substituted alkanoyl, optionally substituted -C(O)-alkyloxy, optionally substituted -C(O)-aryloxy, optionally substituted -C(O)-aralkyloxy, optionally substituted -C(O)-heterocyclyloxy, optionally substituted -C(O)-heteroarylalkyloxy, optionally substituted -C(O)NH-alkyl, optionally substituted -C(O)NH-aryl, optionally substituted -C(O)NH-arylalkyl, optionally substituted -SO2-heterocyclyl, optionally substituted -SO2-alkyl, optionally substituted -SO2-aryl, optionally substituted -SO2-arylalkyl, optionally substituted -SO2-heteroarylalkyl, optionally substituted -SO2-heteroaryl, and optionally substituted ureido); or the first Rx is hydrogen and the second Rx is an amino acid residue Xaa0); and is an N-terminal group of Xaa1 selected from the group consisting of Xaa0 is an optionally substituted amino acid residue selected from the group consisting of Gly, Pro, Arg, Glu, His, Phe, Trp, and Aib; Xaa1 is an optionally substituted amino acid residue containing an amino acid side chain containing an alkyl, aryl or heteroaryl group; X aa 2 is an optionally substituted amino acid residue selected from the group consisting of Gly, Aib, Ala, D-Ala, N-methyl-Ala, N-methyl-D-Ala, Pro, α-methyl-Pro, Val, D-Val, and D-His; X aa 3 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 4 is an amino acid residue selected from the group consisting of Gly, Ala, Aib, and β-Ala; X aa 5 is an optionally substituted amino acid selected from the group consisting of Thr, Ser, Ala, Aib, Val, α-MeSer, α-MeThr, and α-MeVal; X aa 6 is an optionally substituted amino acid residue disubstituted at the α-carbon, and one of the α-carbon substituents is an optionally substituted aryl or an optionally substituted heteroaryl; X aa 7 is an optionally substituted amino acid residue containing an amino acid side chain containing a hydroxyl group; X aa 8 is an optionally substituted amino acid residue selected from the group consisting of Ser, His, and Asn; X aa 9 is an optionally substituted amino acid residue containing an amino acid side chain containing a carboxyl or sulfonic acid group; X aa 10 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; X aa 11 is an optionally substituted amino acid residue containing an amino acid side chain containing a sulfide group, an optionally substituted aryl, an optionally substituted heteroaryl, or any combination thereof; and R YC is the C-terminal group of Xaa 11 having the structure -C(O)N(R Y ) 2, wherein each R Y is independently hydrogen or a PK-modifying group, and the PK-modifying group improves the pharmacokinetic profile of the polypeptide. A composition for use in the treatment or prevention of a neurodegenerative disease or disorder, comprising a polypeptide represented by The treatment or prevention comprises administering the polypeptide to a subject in need thereof, The neurodegenerative disease or disorder is at least partially mediated by glucagon-like peptide 1 (GLP-1), Optionally, the neurodegenerative disease or disorder is selected from Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, Huntington's disease, and prion disease, Optionally, the administering step is by oral administration or parenteral administration, the composition.

25. The following 【Chemical formula 22】 The polypeptide according to claim 1, represented by

26. The following 【Chemical formula 23】 The polypeptide according to claim 1, represented by

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