Improved peptide drugs for insulin resistance

Abstract

To provide safe and effective medications for the treatment of obesity and diabetes mellitus. [Solution] Methods for the synthesis and therapeutic use of covalently modified peptides and / or proteins are described. Covalently modified peptides and / or proteins enable the improvement of the pharmaceutical properties of peptide and protein-based therapeutics. In one embodiment, a peptide product is provided comprising a surfactant X covalently bonded to a peptide, wherein the peptide comprises a linker amino acid U and at least one other amino acid, any two of aa1-aa37 are optionally cyclized via their side chains to form a lactam bond, provided that one or at least one of aa10-aa37 is the linker amino acid U covalently bonded to X.

Description

[Technical Field]

[0001] <Related applications> This application claims the benefits of U.S. Provisional Patent Application No. 62 / 004,156, titled "PEPTIDE PHARMACEUTICALS FOR INSULIN RESISTANCE," filed on 28 May 2014, which is incorporated herein by reference in whole.

[0002] The increasing prevalence of diabetes mellitus vera poses a global health crisis with the potential to become a major epidemic. It is a leading cause of disease and mortality among patients and also represents a significant economic burden. Obesity is a significant risk factor for type 2 diabetes, with approximately 90% of people with type 2 diabetes being overweight or obese. Obesity is a rapidly increasing problem worldwide, and currently, more than 65% of adults in the United States are overweight (Hedley, AA, et al. (2004) JAMA 291:2847-2850). The development of safe and effective drug treatments for obesity and diabetes mellitus vera is urgently needed. [Overview of the project] [Problems that the invention aims to solve]

[0003] This specification describes compositions and methods for the treatment or prevention of diseases associated with insulin resistance, including, but not limited to, obesity, syndrome X, metabolic syndrome, insulin resistance, type 2 diabetes, hypertension, cardioprotection, atherosclerosis, myocardial infarction, and β-cell protection. In some embodiments, the methods include prophylactic and / or therapeutic treatments using peptides and / or proteins. Peptide and / or protein pharmaceuticals have several limitations in their use in medicine—for example, short duration of action, poor bioavailability, and low selectivity for receptor subtypes (Nestor, JJ, Jr. (2007) Comprehensive Medicinal Chemistry II 2:573-601). In addition, peptides and / or proteins are unstable in formulations and are often prone to aggregation.

[0004] This specification describes certain covalently modified peptides and / or proteins (e.g., GLP-1, glucagon, and related analogs) that extend the duration of action after administration and / or improve bioavailability. Such covalently modified peptides and / or proteins are suitable for the prevention and / or treatment of diseases associated with obesity, metabolic syndrome, insulin resistance, type 2 diabetes, hypertension, atherosclerosis, and the like.

[0005] In some embodiments, the covalently modified peptides and / or proteins described herein are bound to a glycosidic surfactant. In one embodiment, the covalently modified peptide and / or protein is bound to a glycosidic surfactant, the peptide and / or protein is bound to a glycoside in the surfactant, and the glycoside is then bound to a hydrophobic group. Similarly, in some embodiments, reagents and intermediates for the synthesis of peptides and / or proteins modified by surfactant incorporation (e.g., modified GLP-1, glucagon, oxytomodulin, or analogs of GLP-1) are also provided. [Means for Solving the Problem]

[0006] In some embodiments, there is provided a peptide product comprising a surfactant X covalently bound to a peptide, wherein the peptide comprises a linker amino acid U and at least one other amino acid:

[0007] [Chemical Formula] wherein the surfactant X is a group of formula I:

[0008] [Chemical Formula] wherein: R 1a is independently, at each occurrence, a single bond, H, a protecting group, a substituted or unsubstituted C1-C 30 alkyl group, a saccharide, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group, R 1b , R 1c , and R 1d are each, at each occurrence, a single bond, H, a protecting group, a substituted or unsubstituted C1-C 30 alkyl group, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group, W 1 is independently, at each occurrence, -CH2-, -CH2-O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH2-S-, W 2 is -O-, -CH2-, or -S-, R 2 is independently, at each occurrence, a single bond to U, H, a substituted or unsubstituted C1-C 30 alkyl group, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group, -NH, -S-, -triazolo-, -NH(C=O)-CH2-, -(CH2) m -maleimide, n is 1, 2, or 3, and, m is an integer between 1 and 10. The peptide was selected from formula II: aa1-aa2-aa3-aa4-aa5-aa6-aa7-aa8-aa9-aa 10 -aa 11 -aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa 18 -aa 19 -aa 20 -aa 21 -aa 22 -aa 23 -aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -aa 30 -aa 31 -aa 32 -aa 33 -aa 34 -aa 35 -aa 36 -aa 37 -Z Formula II (SEQ.ID.NO.1) During the ceremony: Z is OH, NR 4 -His, or -NH-R 3 And, Here, R 3 H, substituted or unsubstituted C1-C 12 Alkyl or PEG chains less than 10 Da, R 4 is C2-C 10 An acyl group, for example, Ac or Bz. aa1 is His, NR 4 -His, pGlu-His, or NR 3 -His is, aa2 is Ser, D-Ser, Ala, Gly, Pro, MePro, Aib, Ac4c, or Ac5c. aa3 is either Gln or Cit. aa4 is Gly or D-Ala, aa5 is either Thr or Ser. aa6 is Phe, Trp, 2FPhe, MePhe, 2FMePhe, or Nal2. aa7 is either Thr or Ser. aa8 is either Ser or Asp. aa9 is either Asp or Glu. aa 10 is Tyr, Leu, Met, Nal2, Bip, Bip2EtMeO, Glu, Lys, or U, aa 11 It does not exist, or it is Ser, Asn, Bip, or U. aa 12 It does not exist, or it is Lys, Glu, Ser, Arg, or U. aa 13 It does not exist, or it is Tyr, Gln, Cit, or U. aa 14 It does not exist, or it is Leu, Met, Nle, Glu, Lys, or U. aa 15 It does not exist, or it is Asp, Glu, or U. aa 16 It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Ac5c, Lys, Arg, or U. aa 17 It does not exist, or it is Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c, Lys, or U. aa 18 It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U. aa 19 It does not exist, or it is Ala, Val, Aib, Ac4c, Ac5c, or U. aa 20 It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U. aa 21It does not exist, or it is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U. aa 22 It does not exist, or it is Phe, Trp, Nal2, Aib, Ac4c, Ac5c, or U. aa 23 It does not exist, or it is Val, Ile, Aib, Ac4c, Ac5c, or U. aa 24 It does not exist, or it is Ala, Gln, Glu, Cit, or U. aa 25 It does not exist, or it is Trp, Nal2, or U. aa 26 It does not exist, or it is Leu, or U. aa 27 It does not exist, or it is Met, Val, Leu, Nle, Lys, or U. aa 28 It does not exist, or it is Asn, Lys, Glu, Gln, Cit, or U. aa 29 It does not exist, or it is Thr, Gly, Aib, Ac4c, Ac5c, or U. aa 30 It does not exist, or it is Lys, Aib, Ac4c, Ac5c, Arg, or U. aa 31 It does not exist, or it is Arg, Aib, Ac4c, Ac5c, or U. aa 32 It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U. aa 33 It does not exist, or it is Arg, Aib, Ac4c, Ac5c, or U. aa 34 It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U. aa 35 It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U. aa 36 It does not exist, or it is Ile, Aib, Ac4c, Ac5C, or U. aa 36is absent, or is Ala, Aib, Ac4c, Ac5C, or U, aa 37 is absent, or is U, U is a natural or non-natural amino acid comprising a functional group used for covalent bonding to surfactant X, here, aa1-aa 37 any two of which are optionally cyclized via their side chains to form a lactam bond, provided that aa 10 -aa 37 one or at least one of is assumed to be a linker amino acid U covalently bonded to X.

[0009] In some embodiments, n is 1. In some embodiments, n is 2, and the first glycoside has a W of the first glycoside 2 and OR of the second glycoside 1b , OR 1c , or OR 1d is bonded to the second glycoside via a bond between any one of . In some embodiments, n is 3, and the first glycoside has a W of the first glycoside 2 and OR of the second glycoside 1b , OR 1c , or OR 1d is bonded to the second glycoside via a bond between any one of , and the second glycoside has a W of the second glycoside 2 and OR of the third glycoside 1b , OR 1c , or OR 1d is bonded to the third glycoside via a bond between any one of .

[0010] In one embodiment, the compound of formula I-A is a compound wherein X has the following structure:

[0011]

Chemical Formula

[0012] In another embodiment, the compound of formula I-A is a compound wherein X has the following structure:

[0013]

Chemical

[0014] Accordingly, in the embodiments described above, R 2 is a single bond.

[0015] For example, in an exemplary embodiment of the structure of X described above, W 1 is -C(=O)NH-, and R 2 is a single bond between W 1 and an amino acid residue U in the peptide (e.g., an amino group in the side chain of a lysine residue in the peptide).

[0016] In a further embodiment, the compound of formula I-A is a compound wherein X has the following structure:

[0017]

Chemical

[0018] For example, in a typical embodiment of the structure of X described above, W 1 is -CH2-, and R 2 R is an alkyl-bonded maleimide functional group on X, 2 It binds to the appropriate portion of amino acid residue U within the peptide (for example, a thiol group in a cysteine ​​residue of a peptide forms a thioether with maleimide on X).

[0019] In yet another embodiment, the compound of formula IA is a compound having the following structure:

[0020] [ka] During the ceremony: R 1a H, protecting group, saccharide, substituted or unsubstituted C1-C 30 This is a portion containing an alkyl group or a steroid nucleus. R 1b , R 1c , and R 1d Each instance independently contains H, a protecting group, or a substituted or unsubstituted C1-C. 30 It is an alkyl group, W 1 is -(C=O)-NH-, W 2 is -O-, R 2 It is a single bond.

[0021] In additional embodiments, the compound of formula IA is a compound in which X has the following structure:

[0022] [ka] During the ceremony: R 1a C1-C is either substituted or non-substituted. 30 It is an alkyl group, R 1b, R 1c , and R 1d H is H, W 1 is -(C=O)-NH-, W 2 is -O-, and, R 2 It is a single bond.

[0023] In some embodiments described above and in this specification, R 1a C1-C is either substituted or non-substituted. 30 It is an alkyl group.

[0024] In some embodiments described above and in this specification, R 1a C6-C is either substituted or unsubstituted. 20 It is an alkyl group.

[0025] In some embodiments described above and in this specification, R 1a is a saccharide. In some embodiments, the saccharide is galactose. In certain embodiments, the saccharide is alpha-linked galactose. In other embodiments, the saccharide is alpha-linked galactopyranose, beta-linked galactopyranose, alpha-linked galacofuranose, or beta-linked galacofuranose.

[0026] In this specification, alternative embodiments in which X in formula IA has the following structure are also considered:

[0027] [ka]

[0028] For example, in a typical embodiment of the structure of X described above, W 1 is -S- and R 2 is C1-C 30 It is an alkyl group, W 2 S is R 1a is, W 2It is a single bond between X and the appropriate portion of amino acid residue U within the peptide (for example, a thiol group in a cysteine ​​residue of a peptide forms a thioether with X).

[0029] In another typical embodiment of the structure of X described above, W 1 is -O-, R 2 is C1-C 30 It is an alkyl group, W 2 is O, R 1a is, W 2 This is a single bond between X and the appropriate portion of amino acid residue U within the peptide (for example, a hydroxyl group in a serine or threonine residue of a peptide forms an ether with X).

[0030] In another typical embodiment of the structure of X described above, W 2 is -O-, R 2 is C1-C 30 It is an alkyl group, W 1 CO is CO, and R 2 This is a spacer amino acid structure, such as Glum or Lysm, that binds to the appropriate portion of amino acid residue U within the peptide (for example, the Glu spacer binds to the epsilon-amino function of Lys in the peptide via its gamma-CO, or Lys binds to the epsilon-amino function of Lys in the peptide via its alpha-CO).

[0031] In some embodiments, U is used for the covalent bond to X and is a dibase natural or unnatural amino acid, a natural or unnatural amino acid containing a thiol, an unnatural amino acid containing a -N3 group, an unnatural amino acid containing an acetylene group, or -NH-C(=O)-CH2-Br or -(CH2) m - A non-natural amino acid containing maleimide, with m being 1-10.

[0032] In some embodiments of the peptide product, the surfactant is a surfactant of the 1-alkyl glycoside class. In some embodiments of the peptide product, the surfactant is bonded to the peptide by an amide bond.

[0033] In some embodiments of the peptide product, surfactant X is 1-eicosyl beta-D-glucuronic acid, 1-octadecyl beta-D-glucuronic acid, 1-hexadecyl beta-D-glucuronic acid, 1-tetradecyl beta-D-glucuronic acid, 1-dodecyl beta-D-glucuronic acid, 1-decyl beta-D-glucuronic acid, 1-decyl beta-D-glucuronic acid, 1-octyl beta-D-glucuronic acid, 1-eicosyl beta-D-diglucuronic acid, 1-octadecyl beta-D-diglucuronic acid Acids, 1-hexadecylbeta-D-diglucuronic acid, 1-tetradecylbeta-D-diglucuronic acid, 1-dodecylbeta-D-diglucuronic acid, 1-decylbeta-D-diglucuronic acid, 1-octylbeta-D-diglucuronic acid, or functionalized 1-eicosylbeta-D-glucose, 1-octadecylbeta-D-glucose, 1-hexadecylbeta-D-glucose, 1-tetradecylbeta-D-glucose, 1-dodecylbeta-D-glucose, 1 -decylbeta-D-glucose, 1-octylbeta-D-glucose, 1-eicosylbeta-D-maltoside, 1-octadecylbeta-D-maltoside, 1-hexadecylbeta-D-maltoside, 1-tetradecylbeta-D-maltoside, 1-dodecylbeta-D-maltoside, 1-decylbeta-D-maltoside, 1-octylbeta-D-maltoside, 1-eicosylbeta-D-melibioside, 1-octadecylbeta-D-maltoside The compounds include ta-D-melibioside, 1-hexadecylbeta-D-melibioside, 1-tetradecylbeta-D-melibioside, 1-dodecylbeta-D-melibioside, 1-decylbeta-D-melibioside, 1-octylbeta-D-melibioside, and other similar compounds, along with the corresponding 6' or 6',6-carboxylic acids. The peptide products are prepared by the formation of bonds between the aforementioned groups and the groups on the peptide (e.g., the COOH group of the aforementioned groups and the amino group of the peptide).In some embodiments, the surfactant X is 1-tetradecylbeta-D-maltoside, 1-dodecylbeta-D-maltoside, 1-decylbeta-D-maltoside, 1-octylbeta-D-maltoside, 1-eicosylbeta-D-melibioside, 1-octadecylbeta-D-melibioside, 1-hexadecylbeta-D-melibioside, 1-tetradecylbeta-D-melibioside, 1-dodecylbeta-D-melibioside, 1-decylbeta-D-melibioside, or 1-octylbeta-D-melibioside, as well as the corresponding 6' or 6',6 carboxylic acid. In some embodiments, the surfactant X is 1-tetradecylbeta-D-maltoside, 1-eicosylbeta-D-melibioside, 1-octadecylbeta-D-melibioside, 1-hexadecylbeta-D-melibioside, 1-tetradecylbeta-D-melibioside, 1-dodecylbeta-D-melibioside, 1-decylbeta-D-melibioside, or 1-octylbeta-D-melibioside.

[0034] In some embodiments of the peptide product, U is a terminal amino acid of the peptide. In some embodiments of the peptide product, U is a non-terminal amino acid of the peptide. In some embodiments of the peptide product, U is a natural D- or L-amino acid. In some embodiments of the peptide product, U is a non-natural amino acid. In some embodiments of the peptide product, U is selected from Lys, Cys, Orn, or a non-natural amino acid containing a functional group used for covalent bonding to surfactant X.

[0035] In some embodiments of the peptide product, the functional groups used for covalent bonding of the peptide to the surfactant X are -NH2, -SH, -OH, -N3, haloacetyl, and -(CH2). m - Maleimide (where m is 1-10), or an acetylene group.

[0036] In some embodiments, the side-chain functional groups of two different amino acid residues are linked to form a cyclic lactam. This linkage is indicated by an asterisk on the two thus linked residues. For example, in some embodiments, Lys * The side chain is Glu * The side chain forms a cyclic lactam. In some embodiments, this lactam structure is reversed, and Glu * and Lys * These are formed from the following. In some cases, these lactam bonds are known to stabilize the alpha-helical structure in the peptide (Condon, SM, et al. (2002) Bioorg Med Chem 10: 731-736; Murage, EN, et al (2008) Bioorg Med Chem 16: 10106-12); Murage, EN, et al. (2010) J Med Chem 53: 6412-20). In some embodiments, cysteine ​​residues may be bonded by disulfide formation to achieve a similar form of conformational constraint and to aid in the formation of the helical structure (Li, Y., et al. (2011) Peptides 32: 1400-1407). In some embodiments, the side-chain functional groups of two different amino acid residues combine to form a heterocycle generated through a "click reaction" between the side-chain azide and alkyne functional groups, thereby achieving a conformational analogue of a stable helical structure with steric constraints (Le Chevalier Isaad A., et al. (2009) J Peptide Sci 15:451-4). In some embodiments, the side-chain functional groups of two different amino acid residues combine to form a CC double bond via the use of olefin metathesis, which may be further modified by reduction to a CC single bond (Verdine, GL and Hilinski, GJ (2011) Meth Enzymol 503:3-33).

[0037] In some embodiments, the peptide product containing a covalently bonded alkyl glycoside is a covalently modified glucagon or an analog thereof. In some of these embodiments, the peptide product contains a covalently bonded 1-O-alkylβ-D-glucuronic acid, and the peptide is an analog of glucagon.

[0038] In some embodiments, the peptide product comprising a covalently bonded alkyl glycoside is a covalently modified GLP-1 or an analog thereof. In some of these embodiments, the peptide product comprises a covalently bonded 1-O-alkylβ-D-glucuronic acid, and the peptide is an analog of GLP-1.

[0039] In some embodiments, the peptide product of formula IA has the structure of formula III-A (SEQ.ID.NO.2), aa1-aa2-aa3-aa4-aa5-aa6-aa7-aa8-aa9-aa 10 -aa 11 -aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa 18 -aa 19 -aa 20 - aa 21 -aa 22 -aa 23 -aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -Z Form III-A (SEQ.ID.NO.2) During the ceremony: Z is either OH or -NH-R 3 And, R 3 H, substituted or unsubstituted C1-C 12 Alkyl or PEG chains less than 10 Da, aa1 is His, N-Ac-His, pGlu-His, or NR 3-His is, aa2 is Ser, Ala, Gly, MePro, Aib, Ac4c, or Ac5c. aa3 is either Gln or Cit. aa4 is Gly or D-Ala, aa5 is either Thr or Ser. aa6 is Phe, Trp, 2FPhe, MePhe, 2FMePhe, or Nal2. aa7 is either Thr or Ser. aa8 is either Ser or Asp. aa9 is either Asp or Glu. aa 10 is Tyr, Leu, Met, Nal2, Bip, Bip2EtMeO, Glu, Lys, or U(X), aa 11 It does not exist, or it is Ser, Asn, Bip, or U(X), aa 12 It does not exist, or it is Lys, Glu, Ser, Arg, or U(X), aa 13 It does not exist, or it is Tyr, Gln, Cit, or U(X), aa 14 It does not exist, or it is Leu, Met, Nle, Glu, Lys, or U(X), aa 15 It does not exist, or it is Asp, Glu, or U(X), aa 16 It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Ac5c, Lys, Arg, or U(X), aa 17 It does not exist, or it is Arg, hArg, Gln, Glu, Lys, Cit, Aib, Ac4c, Ac5c, or U(X), aa 18 It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X), aa 19 It does not exist, or it is Ala, Val, Aib, Ac4c, Ac5c, or U(X), aa 20 It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U(X), aa 21 It does not exist, or it is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U(X), aa 22 It does not exist, or it is Phe, Trp, Nal2, Aib, Ac4c, Ac5c, or U(X), aa 23 It does not exist, or it is Val, Ile, Aib, Ac4c, Ac5c, or U(X), aa 24 It does not exist, or it is Ala, Gln, Glu, Cit, or U(X), aa 25 It does not exist, or it is Trp, Nal2, or U(X), aa 26 It does not exist, or it is Leu, or U(X), aa 27 It does not exist, or it is Met, Val, Leu, Nle, Lys, or U(X), aa 28 It does not exist, or it is Asn, Lys, Glu, Gln, or U(X), aa 29 It does not exist, or it is Thr, Gly, Aib, Ac4c, Ac5c, or U(X), Here, aa1-aa 29 Any two of them can be optionally cyclized via side chains to form a lactam bond. However, aa 10 aa 11 aa 12 aa 16 aa 17 aa 18 aa 19 aa 20 aa 21 aa 22 aa 23 aa 24 aa 25 aa 26 aa 27aa 28 , or aa 29 Assume that one or at least one of these is a natural or unnatural amino acid U covalently bonded to X.

[0040] In some embodiments, the peptide product of formula IA has the structure of formula III-A, aa1-aa2-aa3-aa4-aa5-aa6-aa7-aa8-aa9-aa 10 -aa 11 -aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa 18 -aa 19 -aa 20 - aa 21 -aa 22 -aa 23 -aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -Z Formula III-A((SEQ.ID.NO.2) During the ceremony: Z is either OH or -NH-R 3 And, R 3 H, substituted or unsubstituted C1-C 12 Alkyl or PEG chains less than 10 Da, aa1 is His, aa2 is Aib, aa3 is Gln, aa4 is Gly, aa5 is Thr, aa6 is Phe, aa7 is Thr, aa8 is Ser, aa9 is Asp, aa 10 is Tyr, Glu, Lys, or U(X), aa 11 is Ser, aa 12 Lys and Glu are aa 13 is Tyr, aa 14 These are Leu, Glu, and Lys. aa 15 It is Asp, aa 16 Glu and Lys are, aa 17 is Gln, Glu, or U(X), aa 18 It is Ala, aa 19 It is Ala, aa 20 is Glu, Lys, or U(X), aa 21 is Glu, aa 22 It is Phe, aa 23 is Ile, aa 24 is Gln, Glu, or U(X), aa 25 It is a trump, aa 26 is Leu, aa 27 is Leu, aa 28 It is Glu or Gln, aa 29 is Thr, Here, aa 16 and aa 20 , or aa 10 and aa 14 , or aa 12 and aa 16 It is optionally cyclized by its side chains to form lactam bonds, However, aa 10 aa 17 aa 20 , or aa 24 Assume that one or at least one of them is a natural or unnatural amino acid U covalently bonded to X.

[0041] In some embodiments, the peptide product of formula IA has the structure of formula III-B. His1-aa2-aa3-Gly4-Thr5-aa6-Thr7-Ser8-Asp9-aa 10 -aa 11 -aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa 18 -aa 19 -aa 20 -aa 21 -aa 22 -aa 23 - aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -aa 30 -Z Form III-B (SEQ.ID.NO.3) During the ceremony: Z is either OH or -NH-R 3 And, R 3 H, substituted or unsubstituted C1-C 12 Alkyl or PEG chains less than 10 Da, aa2 is Gly, MePro, or Aib. aa3 is either Gln or Cit. aa6 is Phe, 2FPhe, MePhe, 2FMePhe, or Nal2. aa 10 is Tyr, Nal2, Bip, Bip2EtMeO, Glu, Lys, or U(X), aa 11 It does not exist, or it is Ser, Asn, Bip, or U(X), aa 12 It does not exist, or it is Lys, Glu, Ser, or U(X), aa 13 It does not exist, or it is Tyr, Gln, Cit, or U(X), aa14 It does not exist, or it is Leu, Nle, Glu, Lys, or U(X), aa 15 It does not exist, or it is Asp, Glu, or U(X), aa 16 It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Lys, Arg, or U(X), aa 17 It does not exist, or it is Arg, hArg, Gln, Glu, Lys, Cit, Aib, or U(X), aa 18 It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X), aa 19 It does not exist, or it is Ala, Aib, or U(X), aa 20 It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, or U(X), aa 21 It does not exist, or it is Asp, Glu, Leu, Aib, or U(X), aa 22 It does not exist, or it is Phe, or U(X), aa 23 It does not exist, or it is Val, Ile, Aib, or U(X), aa 24 It does not exist, or it is Ala, Glu, Gln, or U(X), aa 25 It does not exist, or it is Trp, or U(X), aa 26 It does not exist, or it is Leu, or U(X), aa 27 It does not exist, or it is Met, Val, Leu, Nle, Lys, or U(X), aa 28 It does not exist, or it is Asn, Glu, Gln, Cit, or U(X), aa 29 It does not exist, or it is Thr, Aib, or U(X), aa30 It does not exist, or it is Arg, or U(X), Here, aa1-aa 23 Any two of them can be optionally cyclized by their side chains to form a lactam bond. However, aa 10 aa 11 aa 12 aa 16 aa 17 aa 18 aa 19 aa 20 aa 21 aa 22 aa 23 aa 24 , or aa 28 Assume that one or at least one of them is a natural or unnatural amino acid U covalently bonded to X.

[0042] In some embodiments of formulas IA, III-A, or III-B, U is any linker amino acid described herein.

[0043] In some embodiments of formula IA, III-A, or III-B, aa 12 is lysine. In some embodiments of formula IA, III-A, or III-B, aa 14 It is leucine.

[0044] In some embodiments of formula IA, III-A, or III-B, aa 18 This is a lysine residue bound to X.

[0045] In some embodiments of formula IA, III-A, or III-B, aa 17 This is a homoarginine (hArg) residue.

[0046] In some embodiments of formula IA, III-A, or III-B, aa 17 This is a glycine residue.

[0047] In some embodiments of formulas IA, III-A, or III-B, aa2 is a residue of Aib or Ac4c. In some embodiments, aa2 is an Aib residue.

[0048] In some embodiments of formulas IA, III-A, or III-B, the peptide comprises one or more Aib residues.

[0049] In some embodiments of formulas IA, III-A, or III-B, the peptide contains one or more Aib residues at its C-terminus.

[0050] In some embodiments of formulas IA, III-A, or III-B, the peptide includes an amino acid spacer between the surfactant saccharide and the linker amino acid in the peptide.

[0051] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Ala 18 -Ala 19 -aa 20 -Glu 21 -Phe 22 -aa 23 -aa 24 -Trp 25 -Leu 26 -aa 27 -aa 28 -Thr 29 -NH2; (SEQ.ID.NO.774) During the ceremony, aa2 is either Gly or Aib. aa 16 is Glu, Ser, Ala, Lys, or Aib, aa17 is Gln, Glu, Lys, or U(X), aa 20 It is Lys, Glu, or Arg. aa 23 is Ile or Val, aa 24 is Ala, Gln, or U(X), aa 27 It is Met, Val, or Leu, aa 28 This is Asn, Gln, or U(X).

[0052] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2; (SEQ.ID.775) During the ceremony, aa2 is either Gly or Aib. aa 16 These are Glu, Ala, and Aib, aa 17 is Lys, or U(X), aa 27 It is either Leu or Val.

[0053] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Arg 18 -Ala 19 -aa 20 -Asp 21 -Phe 22 -aa 23 -aa 24 -Trp 25 -Leu 26 -aa 27 -aa 28 -Thr 29 -NH2; (SEQ.ID.NO.776) During the ceremony, aa2 is either Gly or Aib. aa 16 is Glu, Ser, Ala, or Aib, aa 17 is Arg, hArg, or Gln, aa 20 is Lys, or U(X), aa 23 It is Ile or Val, aa 24 is Ala, Gln, or U(X), aa 27 is Leu, or Val, and, aa 28 This is Asn, Gln, or U(X). In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Arg 18 -Ala 19 -aa 20 -Asp 21 -Phe 22 -aa 23 -aa 24 -Trp 25 -Leu 26 -aa 27 -aa 28 -Thr 29 -NH2; (SEQ.ID.NO.777) During the ceremony, aa2 is either Gly or Aib. aa 16 These are Glu, Ser, Ala, and Aib. aa 17 is Arg, hArg, or Gln, aa 20 is Lys, or U(X), aa 23 It is Ile or Val, aa 24 is Gln, Ala, or U(X), aa 27 is Leu or Val, aa 28 This is Asn, Gln, or U(X).

[0054] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Ala 18 -Ala19 -aa 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega-X) 24 -Trp 25 -Leu 26 -aa 27 -aa 28 -Thr 29 -NH2; (SEQ.ID.NO.778) During the ceremony, aa2 is either Aib or Gly. aa 16 and aa 20 Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond. aa 17 is Arg, hArg, or Gln, aa 27 It is Met, Val, Leu, or Nle, aa 28 is Asn or Gln, and, Alkyl is C8-C 20 It is a linear alkyl chain. In some embodiments, Lys(N-omega-X) 24 This is Lys(N-omega-1'-alkylbeta-D-glucuronyl).

[0055] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22-Ile 23 -Ala 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2; (SEQ.ID.NO.779) During the ceremony, aa2 is either Aib or Gly. aa 16 It is Glu, Ala, or Aib, aa 17 is Lys or Lys(N-omega-X), and, Alkyl is C8-C 20 It is a linear alkyl chain. In some embodiments, aa 17 This is Lys(N-omega-1'-alkylbeta-D-glucuronyl).

[0056] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Arg 18 -Ala 19 -aa 20 -Asp 21 -Phe 22 -aa 23 -aa 24 -Trp 25 -Leu 26 -aa 27 -aa 28 -Thr 29 -NH2; (SEQ.ID.NO.780) During the ceremony, aa2 is either Gly or Aib. aa16 These are Glu, Ala, and Aib, aa 17 These are Arg and hArg, aa 20 is Lys or Lys(N-omega-X), aa 23 It is Ile or Val, aa 24 It is Gln or Ala, aa 27 is Leu or Val, aa 28 is Asn or Gln, and, Alkyl is C8-C 20 It is a linear alkyl chain. In some embodiments, aa 20 This is Lys(N-omega-1'-alkylbeta-D-glucuronyl).

[0057] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-aa6-Thr7-Ser8-Asp9-aa 10 -aa 11 -Z; (SEQ.ID.NO.781) During the ceremony, aa2 is Gly, Aib, or MePro. aa6 is Phe, 2FPhe, MePhe, or 2FMePhe. aa 10 is Tyr, Nal2, Bip, Bip2Et, or Bip2EtMeO, aa 11 is Lys or Lys(N-omega-X), and, Alkyl is C8-C 20 It is a linear alkyl chain. In some embodiments, aa 11 This is Lys(N-omega-1'-alkylbeta-D-glucuronyl).

[0058] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-U(X) 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -aa 17 -Ala 18 -Ala 19 -aa 20 -Glu 21 -Phe 22 -Ile 23 -aa 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -Z; (SEQ.ID.NO.1025) During the ceremony: Z is either OH or -NH-R 3 And, R 3 It is H or contains a PEG chain less than 10 Da, aa 16 and aa 20 Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond. aa 17 It is Glu or Gln, aa 24 It is Ala, Glu, or Gln, aa 28 It is either Asn or Gln.

[0059] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys12 -Tyr 13 -Leu 14 -Asp 15 -Glu 16 -U(X) 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -NH2; (SEQ.ID.NO.795) During the ceremony, aa2 is either Gly or Aib. aa 28 It is either Asn or Gln.

[0060] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -U(X) 17 -Ala 18 -Ala 19 -aa 20 -Glu 21 -Phe 22 -Ile 23 -aa 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -Z; (SEQ.ID.NO.1026) During the ceremony: Z is either OH or -NH-R 3 And, R 3 It is H or contains a PEG chain less than 10 Da, aa 16 and aa 20 Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond. aa 24 It is Ala, Glu, or Gln, aa 28 It is either Asn or Gln.

[0061] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-aa 10 -Ser 11 -Lys 12 -Tyr 13 -aa 14 -Asp 15 -Ser 16 -aa 17 -Ala 18 -Ala 19 -U(X) 20 -Glu 21 -Phe 22 -Ile 23 -aa 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -Z; (SEQ.ID.NO.1027) During the ceremony: Z is either OH or -NH-R 3 And, R 3 It is H or contains a PEG chain less than 10 Da, aa 10 and aa 14 Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond. aa 17 It is Glu or Gln, aa24 It is Ala, Glu, or Gln, aa 28 It is either Asn or Gln.

[0062] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -aa 12 -Tyr 13 -Gln 14 -Asp 15 -aa 16 -aa 17 -Ala 18 -Ala 19 -U(X) 20 -Glu 21 -Phe 22 -Ile 23 -aa 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -Z; (SEQ.ID.NO.1028) During the ceremony: Z is either OH or -NH-R 3 And, R 3 It is H or contains a PEG chain less than 10 Da, aa 12 and aa 16 Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond. aa 17 It is Glu or Gln, aa 24 It is Ala, Glu, or Gln, aa 28 It is either Asn or Gln.

[0063] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -Gln 17 -Ala 18 -Ala 19 -aa 20 -Glu 21 -Phe 22 -Ile 23 -U(X) 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -Z; (SEQ.ID.NO.1029) During the ceremony, Z is either OH or -NH-R 3 And, R 3 It is either H or a PEG chain less than 10 Da, aa 16 and aa 20 Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond, and, aa 28 is Asn or Gln, X comprises a glucuronyl class portion prepared from 1-alkylbeta-D-glucoside, 1-alkylbeta-D-maltoside, 1-alkylbeta-D-melibioside, or the corresponding alpha-glycoside, where the alkyl is C8-C 20 It is a linear alkyl chain.

[0064] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega-X) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -Z; (SEQ.ID.NO.797) During the ceremony, Z is either OH or -NH-R 3 And, aa 16 and aa 20 It is cyclized by its side chains to form a lactam bond, and, X comprises a glucuronyl class portion prepared from 1-alkylbeta-D-glucoside, 1-alkylbeta-D-maltoside, 1-alkylbeta-D-melibioside, or the corresponding alpha-glycoside, where the alkyl is C8-C 20 It is a linear alkyl chain.

[0065] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu *16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-dodecylbeta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.601) In the formula, Glu * 16 and Lys * 20 It is cyclized by its side chains to form a lactam bond.

[0066] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-tetradecylbeta-D-glucuronil)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.602) In the formula, Glu* 16 and Lys * 20 It is cyclized by its side chains to form a lactam bond.

[0067] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-hexadecylbeta-D-glucuronil)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.603) During the ceremony Glu * 16 and Lys * 20 It is cyclized by its side chains to form a lactam bond.

[0068] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14-Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-octadecylbeta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.604). In the formula, Glu * 16 and Lys * 20 It is cyclized by its side chains to form a lactam bond.

[0069] In some embodiments, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega(1-Octylbeta-D-Melibiouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.630) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 - Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega(1-Dodecylbeta-D-Melibiouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.631) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-tetradecylbeta-D-meribiouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.632) His¹-Aib²-Gln³-Gly⁴-Thr⁵-Phe⁶-Thr⁷-Ser⁸-Asp⁹-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-hexadecyl-beta-D-melibionuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH₂; (SEQ. ID. NO. 633) His¹-Aib²-Gln³-Gly⁴-Thr⁵-Phe⁶-Thr⁷-Ser⁸-Asp⁹-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-octadecyl-beta-D-melibionuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH₂; (SEQ. ID. NO. 634) His¹-Aib²-Gln³-Gly⁴-Thr⁵-Phe⁶-Thr⁷-Ser⁸-Asp⁹-Tyr10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega(1-Hexadecyl Alpha-D-Meriviouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.805) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-tetradecyl alpha-D-meribiouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.819) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-hexadecyl alpha-D-melibionuronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.820) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 Lys(N-omega(1-octadecyl alpha-D-melibionuronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.821) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu* 16 -Lys(N-Omega(1-Dodecyl Alpha-D-Meriviouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-dodecylbeta-D-glucouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1099) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-tetradecyl alpha-D-meribiouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-tetradecylbeta-D-glucouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1100) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp15 -Glu * 16 -Lys(N-Omega(1-Hexadecyl Alpha-D-Meriviouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-hexadecylbeta-D-glucouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1101) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-(13-carboxyl-tridecyloxy)beta-D-glucuronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1102) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp15 -Glu * 16 -Lys(N-omega(1-(15-carboxyl-pentadecyloxy)beta-D-glucuronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1103). His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-(17-carboxyl-heptadecyloxy)beta-D-glucuronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1104) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu* 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-(13-carboxyl-tridecyloxy)beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1105) His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-(15-carboxyl-pentadecyloxy)beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1106), or, His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu *16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-(17-carboxyl-heptadecyloxy)beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.1107).

[0070] In some embodiments of formula IA, III-A, or III-B, aa 16 and aa 20 It is cyclized to form a lactam bond.

[0071] In some embodiments, for any compound of formula IA, III-A, or III-B, X comprises an alkyl chain of dodecyl, tetradecyl, hexadecyl, or octadecyl.

[0072] In some embodiments, the peptide product is a bioactive peptide product that binds to GLCR and / or GLP1R.

[0073] In certain embodiments, the peptide products of formula IA, III-A, or III-B described above and herein have the following structure:

[0074] [ka] In the formula, R 1a C1-C is as shown in Table 1 of Figure 1. 20 It is an alkyl chain, where R' is a peptide as described in Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3, and W of formula IA. 2 is -O-, and W of formula IA 1is -(C=O)NH- and is part of the amide bond to peptide R'. In some of these embodiments, R 1a is C6-C 20 It is an alkyl chain. In some of these embodiments, R 1a is C8-C 20 It is an alkyl chain. In some of these embodiments, R 1a is C 12 -C 20 It is an alkyl chain. In some of these embodiments, R 1a It is a C12-C16 alkyl chain.

[0075] In the embodiments described above, the amino portion of an amino acid and / or peptide R' (e.g., the amino group of an amino acid residue such as lysine, or a lysine residue within peptide R') is used to form a covalent bond with a compound having the following structure:

[0076] [ka] Here, R 1a This refers to C1-C as shown in Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3 above. 20 It is an alkyl chain.

[0077] In such cases, the amino acid residue having an amino portion (e.g., lysine in peptide R') used to form a covalent bond with compound A described above is the linker amino acid U bonded to the surfactant X having the structure of formula A. Accordingly, as an example, see Lys(C) in Table 1 of Figure 1, Table 2 of Figure 2, or Table 3 of Figure 3. 12 ) has the following structure.

[0078] [ka]

[0079] In some embodiments of formulas IA, III-A, or III-B, the bond of surfactant X is linked to linker amino acid U by a spacer amino acid or peptide, which may increase the solubility of the overall structure. Such spacer amino acids or amino acid sequences may also originate from bonded amino groups of Glu or Lys, as illustrated in the following structure, where the peptide sequence is at the top, the spacer is in the middle, and X derived from the surfactant is C 12 It is given as, for example, 1-dodecyl beta-D-glucuronic acid:

[0080] [ka]

[0081] Peptide products of formula IA derived from melibiose-based surfactants are also intended within the scope of the embodiments presented herein. For example, the peptides in Table 1 of Figure 1, Table 2 of Figure 2, Table 3 of Figure 3, or Table 4 of Figure 9 contain a lysine linker amino acid bound to a melibiouronic acid-based surfactant X and have the following structure. Such surfactant-derived structure X may be in an alpha or beta configuration at the anomeric position of the glycoside (beta as shown here):

[0082] [ka]

[0083] Furthermore, peptide products of formula IA derived from maltouronic acid-based surfactants via bonding at one or both carboxylic acid functional groups are also intended within the scope of the embodiments presented herein. Thus, as an example, the peptides in Table 1 of Figure 1, Table 2 of Figure 2, or Table 3 of Figure 3 comprise a lysine linker amino acid bonded to a maltouronic acid-based surfactant X and have the following structure:

[0084] [ka]

[0085] In one embodiment, it will be understood that the compound of formula IA is prepared by attaching lysine to a base X, and then attaching additional amino acid residues and / or peptides bound to the lysine-X compound to obtain the compound of formula IA. Furthermore, it will be understood that other natural or non-natural amino acids described herein are suitable for attachment to surfactant X and suitable for attaching additional amino acids / peptides to obtain the compound of formula IA. In another embodiment, it will be understood that the compound of formula IA is prepared by attaching a full-length or partial-length peptide to a base X, and then optionally attaching additional amino acid residues and / or peptides to obtain the compound of formula IA.

[0086] In certain embodiments, compounds selected from the compounds in Table 1 of Figure 1, Table 2 of Figure 2, or Table 3 of Figure 3 are provided herein.

[0087] Furthermore, pharmaceutical compositions comprising the above-mentioned therapeutically effective amount of the peptide product or an acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient are provided herein.

[0088] In some embodiments of the pharmaceutical composition, the carrier is an aqueous-based carrier. In some embodiments of the pharmaceutical composition, the carrier is a non-aqueous-based carrier. In some embodiments of the pharmaceutical composition, the non-aqueous-based carrier is a solvent such as a hydrofluoroalkane, which may contain submicron anhydrous α-lactose or other excipients.

[0089] Within the scope of the embodiments provided herein, a reaction is envisioned between a peptide containing a linker amino acid U having an amino acid and / or a nucleophile and a group X containing a leaving group or a functional group that can be activated by containing a leaving group, such as a carboxylic acid or other reactive group, which enables the covalent bonding of the amino acid and / or peptide to the surfactant X via the linker amino acid U, providing a peptide product of formula IA.

[0090] Similarly, within the scope of the embodiments provided herein, a reaction is envisioned between a peptide containing a linker amino acid U having an amino acid and / or a nucleophile, a peptide containing a linker amino acid U having a leaving group or a functional group that can be activated by having a leaving group, such as a carboxylic acid or other reactive group, and a group X containing a nucleophile, thereby enabling the covalent bonding of an amino acid and / or peptide to a surfactant X via the linker amino acid U, and providing a peptide product of formula IA.

[0091] In one embodiment, it will be understood that the compound of formula IA is prepared by reacting linker amino acid U with X and subsequently adding further residues to U to obtain the peptide product of formula IA. In another embodiment, it will be understood that the compound of formula IA is prepared by reacting a suitable peptide containing linker amino acid U with X and subsequently adding any further residues to U to obtain the peptide product of formula IA.

[0092] Furthermore, this specification also provides a method for synthesizing the above peptide products, the method comprising the following sequence of steps: (a) A step of coupling the peptide with an intermediate, i.e., the compound of formula IV:

[0093] [ka] During the ceremony: R 1aIndependently, at each generation, a single bond, H, saccharide, leaving group, protecting group, natural or unnatural amino acid, substituted or unsubstituted C1-C are produced. 30 Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, R 1b , R 1c , and R 1d Each of these independently, at each occurrence, consists of a single bond, H, leaving group, protecting group, reversibly protected native or unnatural amino acid, and substituted or unsubstituted C1-C. 30 Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, W 1 These are -CH2-, -CH2-O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH2-S-, W 2 is -O-, -CH2-, or -S-, R 2 Independently, at each occurrence, a single bond to U, H, and a substituted or unsubstituted C1-C 30 Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, -NH, -S-, -triazolo-, -NH(C=O)-CH2-, -(CH2) m -It is maleimide, n is 1, 2, or 3. m is an integer between 1 and 10, the process, and (b) A step of optionally deprotecting the coupled peptide from step (a).

[0094] In some embodiments of the method, each natural or unnatural amino acid is independently a reversibly protected linker amino acid at each occurrence. In some embodiments of the method, each natural or unnatural amino acid is independently a reversibly protected lysine or free lysine at each occurrence.

[0095] In some embodiments of the method, the peptide is a peptide of formula II as described above.

[0096] In some embodiments of the method, n is 1, W 1 is -(C=O)-, R 1a C1-C is either substituted or non-substituted. 30 It is an alkyl group, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group, R 2 This is a reversibly protected lysine in a D- or L- configuration.

[0097] In some embodiments of the method, n is 1, W 1 The equation is -(C=O)-. R 1a C8-C is either substituted or unsubstituted. 30 It is an alkyl group, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group. R 2 This is a reversibly protected lysine in a D- or L- configuration.

[0098] In some embodiments of the method, R 1a These are octyl, decyl, dodecyl, tetradecyl, or hexadecyl groups.

[0099] In some embodiments described above and herein, R 1a is a saccharide. In some embodiments, the saccharide is galactose. In certain embodiments, the saccharide is alpha-linked galactose. In other embodiments, the saccharide is alpha-linked galactopyranose, beta-linked galactopyranose, alpha-linked galacofuranose, or beta-linked galacofuranose.

[0100] In some embodiments of the method, n is 1, W 1 is -(C=O)-NH- or -(C=O)-O-, R 2 C1-C is either substituted or non-substituted. 30 The alkyl hydrophobic group, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group, R 1a These are reversibly protected serine or threonine compounds in a D- or L- configuration.

[0101] In some embodiments of the method, R 2 These are octyl, decyl, dodecyl, tetradecyl, or hexadecyl groups.

[0102] In some embodiments of the method, n is 1, m is 1-6; W 1 It is -CH2-, R 1a C1-C is either substituted or non-substituted. 30 The alkyl hydrophobic group, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group, R 2 -Triazolo-, -NH-, -(CH2) m -Maleimide- is NH-(C=O)-CH2-.

[0103] In some embodiments of formula IV, n is 1, W 1 is -(C=O)-O-, R 2 H is H, R 1a C1-C is either substituted or non-substituted. 30 It is an alkyl hydrophobic group.

[0104] In some embodiments of the method, W 1is -(CH2)O. In some embodiments of the method, n is 1. In some embodiments of the method, n is 2, and the first glycoside is W of the first glycoside. 2 and the OR of the second glycoside 1b , OR 1c , or OR 1d It binds to the second glycoside by binding to one of the following:

[0105] In some embodiments of the method, n is 3, and the first glycoside is W of the first glycoside. 2 and the OR of the second glycoside 1b , OR 1c , or OR 1d The second glycoside is bound by a bond with any one of the following, and the second glycoside is W of the second glycoside. 2 and the OR of the third glycoside 1b , OR 1c , or OR 1d It binds to the third glycoside by binding to one of the following:

[0106] In some embodiments of the method, the compound of formula IV is a reversibly protected N-(ε)-(1'-alkylglucuronyl)-lysine in a D- or L- configuration, and R 1a C1-C is either substituted or non-substituted. 20 It is an alkyl chain, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group.

[0107] In some embodiments of the method, the compound of formula IV is a reversibly protected N-ε-(1'-dodecylβ-D-glucuronyl)-lysine in a D- or L- configuration.

[0108] In some embodiments of the method, deprotection involves the use of a weak acid or weak base treatment. In some embodiments of the method, deprotection involves the use of a strong acid.

[0109] In some embodiments, the method further includes steps of chromatography, desalting of the intermediate by reverse-phase high-performance liquid chromatography, or ion-exchange chromatography of the intermediate.

[0110] The pharmaceutical composition comprises a therapeutically effective amount of the peptide product or an acceptable salt thereof as described above and herein, and at least one pharmaceutically acceptable carrier or excipient.

[0111] Methods for treating insulin resistance-related diseases are provided herein, including the administration of any peptide product or compound described herein to an individual in need.

[0112] Methods for treating diabetes mellitus, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, wound healing, insulin resistance, hyperglycemia, hyperinsulinemia, metabolic syndrome, diabetic complications, high blood concentrations of free fatty acids or glycerol, hyperlipidemia, obesity, hypertriglyceridemia, atherosclerosis, acute cardiovascular syndrome, infarction, ischemia-reperfusion, or hypertension are provided herein, comprising the step of administering to an individual requiring a therapeutically effective amount of the peptide product described above and herein.

[0113] A method for reducing weight gain or causing weight loss is provided herein, comprising the step of administering to an individual requiring a therapeutically effective amount of the peptide product described above and herein.

[0114] A method for treating a mammalian disease characterized by obesity-related insulin resistance or metabolic syndrome is provided herein, comprising the step of administering to an individual in need an amount of the peptide product described above and herein that causes weight loss or improves insulin resistance.

[0115] In some embodiments, the disease being treated is metabolic syndrome (X syndrome). In some embodiments, the disease being treated is diabetes mellitus. In some embodiments, the disease being treated is hyperlipidemia. In some embodiments, the disease being treated is hypertension. In some embodiments, the disease being treated is a vascular disease, including atherosclerosis, or a systemic inflammation characterized by elevated C-reactive protein levels.

[0116] In some embodiments of the method, the effective amount of peptide product administered is about 0.1 μg / kg to about 100.0 μg / kg per day, or 0.01 μg / kg to about 1 mg / kg, or 0.1 μg / kg to about 50 mg / kg. In some embodiments, the peptide product is administered parenterally. In some embodiments, the peptide product is administered subcutaneously. In some embodiments, the method of administering the peptide product is nasal inhalation.

[0117] However, it should be understood that the specific dose levels and frequencies for any particular subject requiring treatment may vary and depend on various factors, including the activity of the particular compound employed, the metabolic stability and duration of action of the compound's administration, age, weight, health status, sex, diet, method and timing of administration, rate of excretion, combination drugs, severity of the particular disease, and the host being treated.

[0118] A method is provided for treating metabolic syndrome or its constituent diseases, comprising the step of administering the above-mentioned therapeutically effective amount of peptide product to a subject who requires it. In some embodiments, the metabolic syndrome disease had progressed to diabetes.

[0119] Similarly, covalently modified GLCR and / or GLP1R-binding peptides or analogs thereof, comprising a hydrophilic group and a hydrophobic group covalently bonded to the hydrophilic group, are provided herein. In certain embodiments, the covalently modified peptide and / or protein product comprises a hydrophilic group which is a saccharide and a C1-C 20It contains a hydrophobic group that is an alkyl chain or aralkyl chain.

[0120] In one embodiment, a method is provided for chemically modifying a molecule by covalent bonding to a surfactant in order to increase or retain the biological activity of the composition or molecule, such as receptor binding or enzymatic activity. In some embodiments, the molecule is a peptide. The method may further include further modifications, including covalent bonding of the molecule in the composition to a polymer such as polyethylene glycol.

[0121] Another embodiment provides a method for reducing or eliminating the immunogenicity of a peptide and / or protein drug by covalently bonding a peptide chain to at least one alkyl glycoside, the alkyl having 1 to 30 carbon atoms.

[0122] Furthermore, a method is provided for treating insulin resistance-related diseases, including, but not limited to, obesity, metabolic syndrome, type 2 diabetes, hypertension, and atherosclerosis, the method comprising the step of administering a drug composition comprising a peptide covalently bonded to at least one alkyl glycoside and delivered to a vertebrate, wherein the alkyl has 1 to 30 carbon atoms, 1 to 20 carbon atoms, or further in the range of 6 to 16 carbon atoms, or 6 to 18 carbon atoms, and the covalent bonding of the alkyl glycoside to the peptide increases the drug's stability, bioavailability, and / or duration of action.

[0123] Furthermore, this specification provides for the use of peptide products described herein (e.g., peptide products of formula IA, formula III-A, or formula III-B) for the manufacture of drugs for the treatment of any disease described herein. [Brief explanation of the drawing]

[0124] [Figure 1]Table 1 in Figure 1 depicts the compounds prepared by the methods described herein. This specification provides sequences SEQ.ID.NO.1-3 and SEQ.ID.NO.774-783, 785-797, and 1025-1029. Furthermore, Table 1 in Figure 1 provides the SEQ.ID numbers for compounds EU-A300 to EU-A425, each having SEQ.ID.NO.4-129 as shown in Table 1 in Figure 1. The compounds in Table 1 in Figure 1 and their respective SEQ.ID.NO.s shown in Table 1 in Figure 1 are incorporated herein into the specification at the time of filing. [Figure 2] Table 2 in Figure 2 depicts the compounds prepared by the methods described herein. This specification provides sequences SEQ.ID.NO.1-3 and SEQ.ID.NO.774-783, 785-797, and 1025-1029. Furthermore, Table 2 in Figure 2 provides the SEQ.ID numbers for compounds EU-A426 to EU-A599, each having SEQ.ID.NO.130-137 as shown in Table 2 in Figure 2. The compounds in Table 2 in Figure 2 and their respective SEQ.ID.NO.s shown in Table 2 in Figure 2 are incorporated herein into the specification at the time of filing. [Figure 3] Table 3 in Figure 3 depicts the compounds prepared by the methods described herein. This specification provides sequences SEQ.ID.NO.1-3 and SEQ.ID.NO.774-783, 785-797, and 1025-1029. Furthermore, Table 3 in Figure 3 provides the SEQ.ID numbers for compounds EU-A700 to EU-A1174, having SEQ.ID.NO.318-773:798-806, respectively, as shown in Table 3 in Figure 3. The compounds in Table 3 in Figure 3 and their respective SEQ.ID.NO.s shown in Table 3 in Figure 3 are incorporated herein into the specification at the time of filing. [Figure 4]Figure 4 shows the X-ray crystal structure of the binding site of the extracellular domain of the GLP-1 receptor (Runge, S., et al. (2008) J Biol Chem 283: 11340-7), illustrating the definitive hydrophobic binding element between the receptor and ligand exendin-4 (Val19*, Phe22*, Trp25*, Leu26*), which is mimicked and replaced by the hydrophobic 1'-alkyl moiety of the surfactant on the peptide of the present invention. In this case, the asterisk indicates that the residue is present in the ligand. [Figure 5] Figure 5 shows the in vivo blood glucose response in db / db mice after subcutaneous injection of the listed amounts of the test compounds of the present invention (EU-A993 and EU-A1023) at t=0, 7 hours. [Figure 6] Figure 6 shows examples of detailed structures of some compounds of the present invention and examples of monosaccharide and disaccharide surfactants modified according to one method of the present invention, in this case, their attachment via the epsilon-amino functional group of the Lys residue at position 24. [Figure 7] Figure 7 shows the structure of EU-A992, which is an example of the structural type of the present invention. [Figure 8] Figure 8 illustrates the time course of concentrations of compounds EU-A993, EU-A1023, and the natural hormone GLP-1(7-36) in human plasma during incubation. This data illustrates the extension of the half-life and protection from proteolysis of the compounds of the present invention. [Figure 9] Table 4 in Figure 9 depicts the compounds prepared by the methods described herein. Table 4 in Figure 9 provides the SEQ.ID numbers for compounds EU-A1575 to EU-A1861, having SEQ.ID.NO. 807-1024 and 1030-1098 as shown in Table 4 in Figure 9. The compounds in Table 4 in Figure 9 and their respective SEQ.ID.NO. shown in Table 4 in Figure 9 are incorporated herein by reference into the specification at the time of filing. [Figure 10] Figure 10 shows the weight loss in DIO mice treated with the typical compound EU-A1024. [Figure 11]Figure 11 shows the changes in body fat mass and lean body mass as measured by NMR in DIO mice treated with the typical compound EU-A1024. [Modes for carrying out the invention]

[0125] This specification describes specific covalently modified peptides and / or proteins with improved pharmaceutical properties. Furthermore, methods for the use of covalently modified peptides and / or proteins for the treatment of diseases associated with obesity and metabolic syndrome are provided herein.

[0126] In some embodiments, the modified peptide and / or protein comprises a peptide and / or protein covalently bonded to a hydrophilic group, a "head" (e.g., a polyol (e.g., saccharide)); the hydrophilic group is covalently bonded to a hydrophobic group ("tail"), thereby generating a surfactant. In some embodiments, the use of a hydrophobic glycoside surfactant (e.g., alkyl glycoside) moiety for the covalent modification of a peptide or protein (e.g., glucagon or GLP-1 related peptide) extends the duration of action of the peptide and / or protein through multiple mechanisms, including the formation of a drug depot at the administration site in the body and binding to a hydrophobic carrier protein. In some embodiments, steric hindrance to the peptide and / or protein structure can prevent protease access to the peptide and / or protein product, thereby preventing proteolysis. In some embodiments, surfactant modification of peptides and / or proteins as described herein (e.g., covalent bonding of alkyl glycoside class surfactants) increases transport across the mucosal barrier. Accordingly, the modifications of peptides and / or proteins described herein, but not limited to, result in desirable effects including protection from proteolysis and delayed migration from the site of administration, thereby extending pharmacokinetic behavior (e.g., circulating t). 1 / 2This leads to an extension of the oral mucosa and an improvement in bioavailability.

[0127] In some embodiments, the improved interaction of a modified peptide and / or protein with its receptor is modified in a beneficial manner by sequence truncation, introduction of constraints, and / or incorporation of steric hindrance. This specification describes novel alkylglycoside reagents that enable the incorporation of both rigidity and steric hindrance in modified peptides and / or proteins. In some embodiments, steric hindrance provides receptor selectivity for the modified peptides and / or proteins described herein. In some embodiments, steric hindrance provides protection from proteolysis.

[0128] Proteins and peptides undergo numerous physical and chemical changes that can affect efficacy and safety. These include aggregation, encompassing dimerization, trimerization, and the formation of higher-order aggregates such as amyloid. Aggregation is a major underlying issue for several potentially adverse effects of peptide and / or protein-based therapies, including loss of efficacy, altered pharmacokinetics, reduced stability or product shelf life, and induction of undesirable immunogenicity. The bioavailability and pharmacokinetics of self-associating peptides can be affected by the size of the aggregates and the ease with which non-covalent intermolecular interactions are disrupted at the subcutaneous site (Maji, SK, et al. (2008) PLoS Biol 6:e17). In some cases, peptides aggregate to form subcutaneous depot formulations that last for more than 30 days. 1 / 2Separation is performed. Such slow dissolution yields desirable effects, such as delivery for one month from a single subcutaneous injection, and as a result, the peptide appears inactive in vivo, leading to very low blood concentrations. Therefore, in some cases, hydrophobic aggregation eliminates the bioavailability and efficacy of the peptide (Clodfelter, DK, et al. (1998) Pharm Res 15: 254-262). The modified peptide products described herein are bound to surfactants and are optionally designed to allow interference or enhancement of aggregation as needed.

[0129] Spontaneously occurring oligosaccharides covalently bound to proteins often lack surfactant properties. In some embodiments, the peptides and / or protein products described herein have covalently bound saccharides and additional hydrophobic groups that give the modified peptide surfactant properties, thereby enabling the modification of the bioavailability, immunogenicity, and / or pharmacokinetic behavior of the surfactant-modified peptide.

[0130] Oligosaccharide-modified proteins and peptides can be incorporated, for example, through enzymatic saccharide or oligosaccharide uptake, as described in Jensen, K.J. and Brask, J. (2005) Biopolymers 80: 747-761 (Gijsen, H.J., et al. (1996) Chem Rev 96: 443-474; Sears, P. and Wong, CH (1998) Cell Mol Life Sci 54: 223-252; Guo, Z. and Shao, N. (2005) Med Res Rev 25: 655-678) or through chemical approaches (Urge, L., et al. (1992) Biochem Biophys Res Commun 184: 1125-1132; Salvador, LA, et al. (1995) Tetrahedron 51: 5643-5656; Kihlberg, J., et al. (1997) Methods Enzymol 289: 221-245; Gregoriadis, G., et al. (2000) Cell Mol Life Sci 57: 1964-1969; Chakraborty, TK, et al. (2005) Glycoconj J 22: 83-93; Liu, M., et al. (2005) Carbohydr Res 340: 2111-2122; Payne, RJ, et al. (2007) J Am Chem Soc 129: 13527-13536; Pedersen,SL,et al. (2010) Chembiochem 11: 366-374). Similar to proteins, peptides have also been modified by glycosylation. (Filira, F., et al. (2003) Org Biomol Chem 1: 3059-3063); (Negri, L., et al. (1999) J Med Chem 42: 400-404); (Negri, L., et al. (1998) Br J Pharmacol 124: 1516-1522); Rocchi, R., et al.)(1987) Int J Pept Protein Res 29:250–261; Filira, F., et al. (1990) Int J Biol Macromol 12:41–49; Gobbo, M., et al.(1992) Int J Pept Protein Res 40:54-61; Urge,L., et al. (1992) Biochem Biophys Res Commun 184: 1125-1132; Djedaini-Pillard, F., et al. (1993) Tetrahedron Lett 34: 2457 - 2460; Drouillat, B., et al.(1997)Bioorg Med Chem Lett 7: 2247-2250; Lohof, E.,et al.(2000) Angew Chem Int Ed Engl 39: 2761-2764; Gruner,SA, et al. (2001) Org Lett 3: 3723-3725; Pean, J., et al. (2001) Biochim Biophys Acta 1541: 150-160; Filira, F., et al. (2003) Org Biomol Chem 1:3059–3063; Grotenbreg, GM, et al. (2004) J Org Chem 69:7851–7859; Biondi, L., et al. (2007) J Pept Sci 13:179–189; Koda, Y., et al. (2008) Bioorg Med Chem 16: 6286-6296; Yamamoto, T., et al. (2009) J Med Chem 52: 5164-5175)。.

[0131] However, the aforementioned attempts have not described additional hydrophobic groups attached to oligosaccharides bound to peptides. Accordingly, modified peptides and / or proteins are provided herein that incorporate hydrophobic groups attached to saccharides and / or oligosaccharides covalently bound to peptides and / or proteins, thereby enabling a balance of bioavailability, immunogenicity, and pharmacokinetic behavior. Accordingly, surfactant reagents containing oligosaccharides and hydrophobic groups that enable covalent modification of peptides and / or proteins, such as glucagon and / or GLP-1 and / or analogs, are also provided herein.

[0132] The use of saccharide-based surfactants covalently bonded to peptides for improving the properties of peptides and / or proteins is provided herein. In some embodiments, the modification of peptides and / or proteins with surfactants as described herein (e.g., covalent bonding of alkyl glycoside class surfactants) increases transport across mucosal barriers. In some embodiments, the covalent bonding of surfactants to peptide and / or protein products reduces or prevents aggregation of peptides and / or proteins. In some embodiments, the covalently modified peptides and / or proteins are covalently modified glucagon or GLP-1 peptides or analogs thereof, which are modified to improve their pharmaceutically and medical properties by covalent modification using alkyl glycoside surfactant moieties. Analogues modified with such surfactants have increased steric hindrance, which inhibits proteolysis, slows uptake, and slows clearance from the body.

[0133] In certain cases, the effects of surfactants are beneficial in terms of the physical properties or performance of pharmaceutical formulations, but they are irritating to the skin and / or other tissues, particularly to mucous membranes such as those found in the nose, mouth, eyes, vagina, rectum, buccal, or sublingual regions. Furthermore, in some cases, surfactants denature proteins, disrupting their biological functions. Because surfactants exert an effect beyond the critical micelle concentration (CMC), surfactants with a low CMC are desirable, and as a result, surfactants may be efficiently utilized at low concentrations or in small amounts in pharmaceutical formulations. Accordingly, in some embodiments, surfactants suitable for peptide modification described herein (e.g., alkyl glycosides) have a CMC of less than about 1 mM in pure water or aqueous solution. As an example, the specific CMC values ​​of alkyl glycosides in water are as follows: octyl maltoside 19.5 mM; decyl maltoside 1.8 mM; dodecyl-β-D-maltoside 0.17 mM; tridecyl maltoside 0.03 mM; tetradecyl maltoside 0.01 mM; sucrose dodecanoate 0.3 mM. Appropriate surfactants will be found to have higher or lower CMCs, depending on the peptide and / or protein being modified. As used herein, “critical micelle concentration” or “CMC” is the concentration of an amphiphilic component (alkyl glycoside) in a solution at which the formation of micelles (spherical micelles, rod-shaped micelles, lamellar structures, etc.) in the solution begins. In certain embodiments, alkyl glycosides dodecyl, tridecyl, and tetradecyl maltosides or glucosides, like sucrose dodecanoates, tridecanoates, and tetradecanoates, have a low CMC and are suitable for the modification of peptides and / or proteins described herein.

[0134] Insulin resistance Risks associated with long-term hyperglycemia include increased risk of microvascular complications, sensory neuropathy, myocardial infarction, stroke, macrovascular mortality, and all-cause mortality. Type 2 diabetes is also inevitably linked to obesity, which is a global epidemic. In 2007, at least $232 billion was spent worldwide on treating and preventing diabetes, three-quarters of which was spent in developed countries on general care, such as treating long-term complications and efforts to prevent macrovascular and microvascular complications. In 2007, the estimated indirect cost of diabetes (diabetic disability, loss of productivity, and premature death) to the U.S. economy was $58 billion.

[0135] Obesity causes insulin resistance, which is a decrease in the number of insulin receptors and a reduction in the coupling of these receptors to a critical intracellular signaling pathway, leading to a decreased ability of the body's cells to respond to insulin stimulation. The obese state further causes "metabolic syndrome," a constellation of diseases (such as insulin resistance, hypertension, and atherosclerosis) with very serious healthcare consequences. If insulin resistance is diagnosed sufficiently early, overt type 2 diabetes can be avoided or delayed through lifestyle interventions aimed at reducing calorie intake and body fat, and drug therapy to standardize glycemic control. Despite treatment guidelines recommending early, aggressive intervention, many patients fail to reach their glycemic control targets. Many factors, including psychosocial and economic impacts and shortcomings in the efficacy, ease of use, and tolerability profiles of available antidiabetic drugs, contribute to the failure of type 2 diabetes management. The peptides and / or protein products described herein are intended to overcome these shortcomings.

[0136] Incretin effect The term "incretin effect" is used to describe the phenomenon in which orally delivered glucose loads produce much greater insulin secretion than the same glucose load administered intravenously. This effect is mediated by at least two incretin hormones secreted by intestinal L cells. Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) have been identified as incretins, and it is thought that healthy individuals may derive up to 70% of their dietary insulin secretion response from the incretin effect.

[0137] Normally, incretin peptides are secreted as needed in response to ingested nutrients and have a short plasma half-life due to degradation by the enzyme dipeptidyl peptidase IV (DPP-4). In people with type 2 diabetes, pancreatic responsiveness to GLP-1 is impaired, but insulin secretion response can be restored with pharmacologically administered doses of human GLP-1 (Kieffer, TJ, et al. (1995) Endocrinology 136: 3585-3596). In addition, GLP-1 promotes the regeneration and retention of β-cells (Aaboe, K., et al. (2008) Diabetes Obes Metab 10:994-1003). GLP-1 also has further beneficial effects on cardiac function, etc. (Treiman, M., et al. (2010) Trends Cardiovasc Med 20: 8-12): For example, it improves left ventricular function in human subjects (Sokos, GG, et al. (2006) J Card Fail 12: 694-699). GLP-1 also slows gastric emptying and reduces appetite in humans (Toft-Nielsen, MB, et al. (1999) Diabetes Care 22: 1137-1143).

[0138] Treatment of diabetic patients with metabolically stable, long-acting analogs of GLP-1 is described, for example, in Drab, SR (2010) Pharmacotherapy 30: 609-624, and has issues related to ease of use and side effects such as nausea, pancreatitis, and thyroid cancer. GLP-1 analogs provide glucose-dependent stimulation of insulin secretion and reduce the risk of hypoglycemia. In addition, while many modern diabetes treatments cause weight gain, GLP-1 analogs induce satiety and mild weight loss, as described below. Accordingly, in some embodiments, GLP-1 analogs that are long-acting and administered at low doses, thereby reducing the side effects associated with modern treatments, are provided herein.

[0139] Many peptide gastrointestinal hormones are known to regulate appetite (Sanger, GJ and Lee, K. (2008) Nat Rev Drug Discov 7: 241-254). Some peptides are derived from tissue-specific enzymatic processing (prohormone converters; PCs) of preproglucagon gene products: e.g., glucagon, GLP-1, glucagon-like peptide-2 (GLP-2), glycentin, and oxintomodulin (OXM) (Drucker, DJ (2005) Nat Clin Pract Endocrinol Metab 1: 22-31; Sinclair, EM and Drucker, DJ (2005) Physiology (Bethesda) 20: 357-365). GLP-1, GLP-2, glycentin, and OXM are co-secreted from L cells in the intestine in response to feeding. Preproglucagon is alternatively processed (PC2) in alpha cells in pancreatic islets to produce glucagon. The structure of OXM is essentially glucagon containing an eight-residue C-terminal elongation.

[0140] In addition to stimulating insulin biosynthesis and glucose-dependent insulin secretion, GLP-1 and its stable mimics (e.g., exendin-4, liraglutide) induce moderate weight loss in animal models (Mack, CM, et al. (2006) Int J Obes (Lond) 30: 1332-1340; Knudsen, LB (2010) Int J Clin Pract 64 (Suppl. 167): 4-11) and patients with type 2 diabetes (DeFronzo, RA, et al. (2005) Diabetes Care 28: 1092-1100; Buse, JB, et al. (2010) Diabetes Care 33: 1255-1261). Glucagon infusion reduces human food intake (Geary, N., et al. (1992) Am J Physiol 262: R975-980), while continuous glucagon treatment of adipose tissue further promotes lipolysis (Heckemeyer, CM, et al. (1983) Endocrinology 113: 270-276) and weight loss (Salter, JM, et al. (1960) Metabolism 9: 753-768; Chan, EK, et al. (1984) Exp Mol Pathol 40: 320-327). Glucagon has broad effects on energy metabolism (Heppner, KM, et al. (2010) Physiol Behav). Glucagon or its analogs can be used as a diagnostic technique for temporary paralysis of the gastric tube. Therefore, at least two of the products from the PC treatment of preproglucagon protein are associated with satiety and metabolic effects.

[0141] In rodents, repeated intraperitoneal administration of OXM, a third product of preproglucagon, has been associated with a reduction in white adipose tissue and weight loss compared to controls (Dakin, CL, et al. (2004) Endocrinology 145: 2687-2695). Oxm reduced food intake by 19.3% during intravenous administration to normal-weight humans, and this effect lasted for more than 12 hours after infusion (Cohen, MA, et al. (2003) J Clin Endocrinol Metab 88: 4696-4701). Treatment of volunteers over four weeks maintained satiety and weight loss, reflecting a reduction in body fat (Wynne, K., et al. (2005) Diabetes 54: 2390-2395).

[0142] OXM is structurally homologous to GLP-1 and glucagon, and activates both the glucagon receptor (GCGR) and the GLP-1 receptor (GLP1R), but with 10-100 times less potency than the eponymous ligand from which it derives its name. In addition, studies on the interaction of OXM with GLP1R suggest that it may have a different effect on beta-arrestin supplementation compared to GLP-1 (Jorgensen, R., et al. (2007) J Pharmacol Exp Ther 322: 148-154), and therefore may act as a "biased" ligand. The receptor specific to OXM has long been sought but has not yet been elucidated, and is hypothesized to act via the GLP1R and GCGR pathway. Accordingly, methods for modifying intestinal peptide surfactants that induce satiety, weight loss, mitigation of insulin resistance, and / or delay progression from prediabetes to diabetes are provided herein.

[0143] GLP-1 Given the complex and interactive behavior of preproglucagon protein products on satiety and metabolism described above, researchers from multiple groups have studied the structure-activity relationship between GLP-1 and the glucagon structure. It has been shown that residues throughout the sequence are accepting substitutions. For example, substitutions by Ala are widely accepted in the N-terminal region of GLP-1, particularly at 2, 3, 5, 8, 11, and 12 (Adelhorst, K., et al. (1994) J Biol Chem 269:6275-6278).

[0144] It has been shown that chimeric analogs capable of binding to GLP1R and GLCR can be achieved by grafting a C-terminal residue from GLP-1 onto the N-terminal group of glucagon (Hjorth, SA, et al. (1994) J Biol Chem 269:30121-30124). The residue at position 3 (acidic Gln from GLP1, or neutral Gln in glucagon or OXM) reduces the affinity of glucagon (Runge, S., et al. (2003) J Biol Chem 278:28005-28010) or OXM (Pocai, A., et al. (2009) Diabetes 58: 2258-2266) to GLP1R. The effects on the metabolic profiles of animals treated with stable analogues of GLP-1, glucagon, or OXM having glutamine at position 3 have been studied (Day, JW, et al. (2009) Nat Chem Biol 5: 749-757; Druce, MR, et al. (2009) Endocrinology 150: 1712-1722; Pocai, A., et al. (2009) Diabetes 58: 2258-2266). These analogues were designed to have agonist activity against both GLP1R and GCGR (Day, JW, et al. U.S. Patent No. 2010 / 0190701 A1; Patterson, JT, et al. (2011) J Pept Sci 17: 659-666; Ribier, D., U.S. Patent Application No. 2012 / 0178670).

[0145] Chimeric analogs must have a desirable effect on the parent hormone acting on its receptor, and therefore an effect similar to that of OXM, which at first glance seems to be linked to both GLP-1R and GLCR: glucose-dependent insulin secretion and satiety, and to lipolysis and increased fat burning by glucagon. Analogs have been shown to produce the desirable effects of weight loss and increased fat burning. While such a profile is attractive for the treatment of obesity, a major problem in obesity treatment is compliance. Full-length analogs of glucagon and OXM, respectively, that have affinity for both GLP-1R and GLCR, can result in weight loss, but these analogs are not optimized for the high bioavailability, pharmaceutical properties, and convenient delivery to patients required for an optimal drug therapy regimen. Accordingly, analogs of intestinal peptides (e.g., GLP, OXM, glucagon, etc.) that enable high bioavailability and / or long-lasting effects to improve treatment outcomes in the treatment of diseases such as obesity and / or diabetes and / or metabolic syndrome are provided herein.

[0146] Additional factors in the optimized treatment of metabolic syndrome and diabetes using molecules like OXM relate to the duration of treatment and the amount of glucagon activity. For example, continuous treatment with analogues that activate GLP-1 and the glucagon receptor (OXM pharmacological profile) can result in very large and rapid loss of fat mass (Day, JW, et al. (2009) Nat Chem Biol 5:749-757), but can also cause loss of lean muscle mass (Kosinski, JR, et al. (2012) Obesity (Silver Spring):doi:10.1038 / oby.2012.67), which is undesirable for this class of drugs. For example, in a study by Kosinski, JR et al., the natural hormone OXM was administered continuously for 14 days via Alzet's minipump, resulting in a 30% reduction in fat mass, but also a 7% reduction in lean (muscle) mass.

[0147] While glucagon activity is known to stimulate glycogenolysis, lipolysis, and increased fat burning, it can also have catabolic effects on muscle. Optimal treatments using drugs combining GLP-1 with glucagon activity (OXM profile) require optimally eliciting enhanced glucose-dependent insulin secretion and satiety from the GLP-1 analog, along with appropriate amounts of glucagon activity (fat burning). Furthermore, intermittent use of such drugs provides a desirable clinical profile of moderate, continuous weight loss through loss of body fat, minimizing the loss of lean body mass. This specification refers to molecules with a desirable combination of GLP-1 and OXM activity, as well as an adjustable pharmacokinetic / pharmacokinetic profile, to enable optimal use in the treatment of conditions such as metabolic syndrome, diabetes, and obesity.

[0148] In one embodiment, compounds of formulas IA, III-A, and III-B are designed to yield either glucagon-like activity or GLP-1-like activity. In further embodiments, compounds of formulas IA, III-A, and III-B yield tunable activity. For example, in one example, the peptide products described herein (e.g., compounds in Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3) have an EC50 of less than about 500 nM, preferably less than about 50 nM, and more preferably less than about 20 nM, in receptors for both glucagon and GLP-1. In another example, the peptide products described herein (e.g., the compounds in Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3) are more potent against the GLP-1 receptor (e.g., EC50 less than 10 nM, preferably less than 5 nM, more preferably less than 1 nM) and less potent against the glucagon receptor (e.g., EC50 less than 50 nM, preferably less than about 20 M, more preferably less than about 5 nM). This synchronization of bioactivity allows for the retention of some glucagon activity in reasonable amounts, thereby enabling fat burning, while retaining the beneficial effects of further enhanced glucose-dependent insulin secretion. Structurally, OXM is consistent with GLP-1 and glucagon and activates both the glucagon receptor (GCGR) and the GLP-1 receptor (GLP1R). Therefore, in some embodiments, the compounds of formulas IA, III-A, and III-B result in tunable OXM-like bioactivity. In some specific embodiments, the peptide products described herein include peptides having amino acid residues 1–17 of GLP-1 and / or analogs thereof (e.g., analogs including modified non-natural amino acid substitutions as described herein, cyclized lactam bonds as described herein, modifications with surfactants as described herein, or combinations thereof).In several other embodiments, the peptide products described herein include peptides having amino acid residues 1-16 of GLP-1 and / or analogs thereof (e.g., analogs including modified non-natural amino acid substitutions as described herein, cyclized lactam bonds as described herein, modifications with surfactants as described herein, or combinations thereof). In additional embodiments, the peptide products described herein include peptides having amino acid residues 1-18 of GLP-1 and / or analogs thereof (e.g., analogs including modified non-natural amino acid substitutions as described herein, cyclized lactam bonds as described herein, modifications with surfactants as described herein, or combinations thereof). In addition, the peptide products described herein include one or more residues (e.g., Aib, Ac4c), which result in helix stabilization of the designed compounds of formulas IA, III-A, and III-B, and the compounds of Table 1 in Figure 1, Table 2 in Figure 2, and Table 3 in Figure 3.

[0149] The glucagon subfamily of ligands is thought to bind to many class B receptors (secretin classes, G protein-coupled receptors (GPCRs)) in two domain forms common to these receptors. For GLP-1, the N-terminal region is thought to exist from residue 1 to approximately 16, which binds to the upper part of the intramembrane-extramembrane helix (the region near the membrane) and to the helical C-terminal region of residues 17 to 31, which binds to the large extracellular N-terminal extension (ECD) of the receptor. Such ligand binding highlights the fact that shortened N-terminal analogs of such peptide ligands can still retain substantial binding affinity and selectivity to the isolated ECD region of the receptor. Therefore, it has been suggested that the N-terminal region is responsible for receptor activation, while the C-terminal region is responsible for binding. Recent studies have suggested that short N-terminal analogs of GLP-1 may be promising as both binding agents and receptor activators (Mapelli, C., et al. (2009) J Med Chem 52:7788-7799; Haque, TS, et al. (2010) Peptides 31:950-955; Haque, TS, et al. (2010) Peptides 31:1353-1360).

[0150] In addition, a study of the X-ray crystal structure of the N-terminal region of GLP1R, including the GLP-1 mimetic and a shortened antagonist analog of exendin-4 (Byetta) bound in this region (Runge, S., et al. (2008) J Biol Chem 283:11340-7), shows that the critical ligand-binding region in ECD is highly hydrophobic (Figure 3). The sequence of exendin-4 beyond Glu15 is precisely this hydrophobic region (Val 19* , Phe 22* , Trp 25* , Leu 26*They interact as amphiphilic helices with the asterisk, where the asterisk indicates a residue in the ligand. In one embodiment, a shortened N-terminal fragment of GLP-1 or glucagon is modified to bind to GLCR and covalently bonded to a surfactant. The hydrophobic 1'-alkyl moiety of the surfactant mimics and substitutes the C-terminal region of the natural hormone ligand, increasing the peptide efficacy, effect, and duration of action. In addition, such analogs have a major advantage due to their considerably smaller size, which reduces their complexity, synthesis cost, and susceptibility to proteolysis. Furthermore, even smaller peptides are more readily absorbed through the barrier of intestinal cells in the nasal mucosa or intestinal tract.

[0151] Hypoglycemia is a life-threatening condition characterized by low blood glucose levels, and is becoming increasingly recognized as more aggressive treatments for hyperglycemia, such as intensive insulin therapy, are used in more patients. Hypoglycemia occurs when blood glucose levels fall so low that the brain and muscles cannot receive enough energy for bodily activity. Glucagon may be used to treat this condition, which is achieved by stimulating the liver to break down glycogen and produce glucose, thereby raising blood glucose levels to normal. Analogues of glucagon that retain the ability to activate GLCR may be used to achieve this desired effect on blood glucose levels.

[0152] GLP-1 analogs that activate GLP1R stimulate insulin production from the pancreas and its release in the presence of high blood glucose levels. As seen in current products such as exenatide (Byetta®), this action results in effective control and normalization of blood glucose levels. In addition, such products appear to suppress appetite and slow the movement of food from the stomach. Therefore, they are effective in treating diabetes through multiple mechanisms. Analogues that combine the effects of glucagon and GLP-1, activating both GLCR and GLP1R, may offer benefits in treating diabetes through synergy by suppressing appetite, releasing insulin in a glucose-dependent manner, supporting protection from hypoglycemia, and promoting fat burning.

[0153] Such methods for treating hyperglycemia, including diabetes mellitus, type 1 diabetes mellitus, type 2 diabetes mellitus, or gestational diabetes mellitus, whether insulin-dependent or non-insulin-dependent, are expected to be useful in reducing diabetic complications, including nephropathy, retinopathy, and vascular disease. Applications to cardiovascular disease include both microvascular and macrovascular diseases (Davidson, MH, (2011) Am J Cardiol 108[suppl]:33B-41B; Gejl, M., et al. (2012) J Clin Endocrinol Metab 97:doi:10.1210 / jc.2011-3456), and also include the treatment of myocardial infarction. Such methods for reducing appetite or promoting weight loss are expected to be useful in reducing weight, preventing weight gain, or treating obesity of various causes, including drug-induced obesity, and in reducing complications associated with obesity, including vascular diseases (coronary artery disease, stroke, peripheral vascular disease, ischemia-reperfusion, etc.), hypertension, the development of type II diabetes, hyperlipidemia, and musculoskeletal disorders.

[0154] As used herein, the terms glucagon or GLP-1 analogs include all pharmaceutically acceptable salts or esters thereof.

[0155] <Peptides and their analogues> In one aspect, peptides that are covalently modified and suitable for the methods described herein include, but are not limited to, shortened analogs of glucagon and / or related hormones GLP-1: Glucagon: His1-Ser2-Gln3-Gly4-Thr5Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Ser 16 -Arg 17 -Arg 18 -Ala 19 -Gln 20 -Asp 21 -Phe 22 -Val 23 -Gln 24 -Trp 25 -Leu 26 -Met 27 -Asn 28 -Thr 29 (SEQ.ID.NO.782) Oxinmodulin: His1-Ser2-Gln3-Gly4-Thr5Phe6- Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Ser 16 -Arg 17 -Arg 18 -Ala 19 -Gln 20 -Asp 21 -Phe 22 -Val 23 -Gln 24 -Trp 25 -Leu 26 -Met 27 -Asn 28 -Thr 29 -Lys 30 -Arg 31 -Asn32 -Arg 33 -Asn 34 -Asn 35 -Ile 36 -Ala 37 (SEQ.ID.NO.783) GLP-1 (using glucagon numbering): His1-Ala2-Glu3-Gly4-Thr5Phe6-Thr7-Ser8- Asp9-Val10-Ser 11 -Ser12-Tyr 13 -Leu 14 -Glu 15 -Gly 16 -Gln 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22 -Ile 23 -Ala 24 -Trp 25 -Leu 26 -Val 27 -Lys 28 -Gly 29 -Arg 30 (SEQ.ID.NO.1)

[0156] In some embodiments, the peptide products described herein have the following structure: aa1-aa2-aa3-aa4-aa5-aa6-aa7-aa8-aa9-aa 10 -aa 11 -aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa 18 -aa 19 -aa 20 -aa 21 -aa 22 -aa 23 -aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -aa30 -aa 31 -aa 32 -aa 33 -aa 34 -aa 35 -aa 36 -aa 37 -Z Formula II (SEQ.ID.NO.1) During the ceremony: Z is OH, NR 4 -His, or -NH-R 3 And here, R 3 H, C1-C 12 A substituted or unsubstituted alkyl, or a PEG chain less than 10 Da; R 4 is C2-C 10 An acyl group, for example, Ac or Bz; aa1 is His, NR 4 -His, pGlu-His, or NR 3 -His is; aa2 is Ser, D-Ser, Ala, Gly, Pro, MePro, Aib, Ac4c, or Ac5c; aa3 is Gln or Cit; aa4 is Gly or D-Ala; aa5 is either Thr or Ser; aa6 is Phe, Trp, 2FPhe, MePhe, 2FMePhe, or Nal2; aa7 is either Thr or Ser; aa8 is either Ser or Asp; aa9 is either Asp or Glu; aa 10 is Tyr, Leu, Met, Nal2, Bip, Bip2EtMeO, or U; aa 11 It does not exist, or it is Ser, Asn, Bip, or U; aa 12 It does not exist, or it is Lys, Glu, Ser, Arg, or U; aa 13 It does not exist, or it is Tyr, Gln, Cit, or U; aa 14It does not exist, or it is Leu, Nle, or U; aa 15 It does not exist, or it is Asp, Glu, or U; aa 16 It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Ac5c, Lys, Arg, or U; aa 17 It does not exist, or it is Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c, Lys, or U; aa 18 It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U; aa 19 It does not exist, or it is Ala, Val, Aib, Ac4c, Ac5c, or U; aa 20 It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U; aa 21 It does not exist, or it is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U; aa 22 It does not exist, or it is Phe, Trp, Nal2, Aib, Ac4c, Ac5c, or U; aa 23 It does not exist, or it is Val, Ile, Aib, Ac4c, Ac5c, or U; aa 24 It does not exist, or it is Ala, Gln, Glu, Cit, or U; aa 25 It does not exist, or it is Trp, Nal2, or U; aa 26 It does not exist, or it is Leu or U; aa 27 It does not exist, or it is Met, Val, Leu, Nle, Lys, or U; aa 28 It does not exist, or it is Asn, Lys, Gln, Cit, or U; aa 29 It does not exist, or it is Thr, Gly, Aib, Ac4c, Ac5c, or U; aa 30 It does not exist, or it is Lys, Aib, Ac4c, Ac5c, Arg, or U; aa 31 It does not exist, or it is Arg, Aib, Ac4c, Ac5c, or U; aa 32 It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U; aa 33 It does not exist, or it is Arg, Aib, Ac4c, Ac5c, or U; aa 34 It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U; aa 35 It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U; aa 36 It does not exist, or it is Ile, Aib, Ac4c, Ac5c, or U; aa 36 It does not exist, or it is Ala, Aib, Ac4c, Ac5c, or U; aa 37 It either does not exist or is U; U is a natural or non-natural amino acid that contains a functional group used for covalent bonding to surfactant X; Here, aa1-aa 37 Any two of them can be optionally cyclized via their side chains to form a lactam bond; and aa 10 -aa 37 Assume that one or at least one of them is a linker amino acid U covalently bonded to X.

[0157] In a specific embodiment, the bound amino acid U is a diamino acid such as Ly or Orn, X is a modified surfactant from the class of 1-alkyl glycosides bound to U, Z is OH, or -NH-R2, and R 3is H or C1-C 12 or PEG chains less than 10 Da.

[0158] In some embodiments, the peptide product of formula IA has the following structure of formula III-A: aa1-aa2-aa3-aa4-aa5-aa6-aa7-aa8-aa9-aa 10 -aa 11 -aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa 18 -aa 19 -aa 20 -aa 21 -aa 22 -aa 23 -aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -Z Form III-A (SEQ.ID.NO.2) During the ceremony: Z is OH, -NH-R 3 And here, R 3 is H, or C1-C 12 A substituted or unsubstituted alkyl, or a PEG chain less than 10 Da; aa1 is His, N-Ac-His, pGlu-His, or NR 3 -His is; aa2 is Ser, Ala, Gly, MePro, Aib, Ac4c, or Ac5c; aa3 is Gln or Cit; aa4 is Gly or D-Ala; aa5 is either Thr or Ser; aa6 is Phe, Trp, 2FPhe, MePhe, 2FMePhe, or Nal2; aa7 is either Thr or Ser; aa8 is either Ser or Asp; aa9 is either Asp or Glu; aa 10 is Tyr, Leu, Met, Nal2, Bip, Bip2EtMeO, or U(X); aa 11 It does not exist, or it is Ser, Asn, Bip, or U(X); aa 12 It does not exist, or it is Lys, Glu, Ser, Arg, or U(X); aa 13 It does not exist, or it is Tyr, Gln, Cit, or U(X); aa 14 It does not exist, or it is Leu, Nle, or U(X); aa 15 It does not exist, or it is Asp, Glu, or U(X); aa 16 It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Ac5c, Lys, Arg, or U(X); aa 17 It does not exist, or it is Arg, hArg, Gln, Glu, Lys, Cit, Aib, Ac4c, Ac5c, or U(X); aa 18 It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X); aa 19 It does not exist, or it is Ala, Val, Aib, Ac4c, Ac5c, or U(X); aa 20 It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U(X); aa 21 It does not exist, or it is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U(X); aa 22 It does not exist, or it is Phe, Trp, Nal2, Aib, Ac4c, Ac5c, or U(X); aa 23It does not exist, or it is Val, Ile, Aib, Ac4c, Ac5c, or U(X); aa 24 It does not exist, or it is Ala, Gln, Glu, Cit, or U(X); aa 25 It does not exist, or it is Trp, Nal2, or U(X); aa 26 It does not exist, or it is Leu or U(X); aa 27 It does not exist, or it is Met, Val, Leu, Nle, Lys, or U(X); aa 28 It does not exist, or it is Asn, Lys, Gln, or U(X); aa 29 It does not exist, or it is Thr, Gly, Aib, Ac4c, Ac5c, or U(X); Here, aa1-aa 29 Any two of them can be optionally cyclized via their side chains to form a lactam bond; and aa 10 aa 11 aa 12 aa 16 aa 17 aa 18 aa 19 aa 20 aa 21 aa 22 aa 23 aa 24 aa 25 aa 26 aa 27 aa 28 , or aa 29 Assume that one or at least one of these is a natural or unnatural amino acid U that is covalently bonded to X.

[0159] In some embodiments, the peptide product of formula IA has the following structure of formula III-A: aa1-aa2-aa3-aa4-aa5-aa6-aa7-aa8-aa9-aa 10 -aa 11-aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa 18 -aa 19 -aa 20 -aa 21 -aa 22 -aa 23 -aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -Z Form III-A (SEQ.ID.NO.2) During the ceremony: Z is OH, -NH-R 3 And here, R 3 is H, or C1-C 12 A substituted or unsubstituted alkyl, or a PEG chain less than 10 Da; aa1 is His; aa2 is Aib; aa3 is Gln; aa4 is Gly; aa5 is Thr; aa6 is Phe; aa7 is Thr; aa8 is Ser; aa9 is Asp; aa 10 is Tyr, Glu, Lys, or U(X); aa 11 is Ser; aa 12 It is Lys or Glu; aa 13 is Tyr; aa 14 It is Leu, Glu, or Lys; aa 15 is Asp; aa 16 is either Glu or Lys; aa 17 is Gln, Glu, or U(X); aa 18 is Ala; aa 19 is Ala; aa 20 is Lys, Glu, or U(X); aa 21 is Glu; aa 22 is Phe; aa 23 is Ile; aa 24 is Gln, Glu, or U(X); aa 25 is a trump; aa 26 is Leu; aa 27 is Leu; aa 28 is Glu or Gln; aa 29 is Thr; Here, aa 16 and aa 20 , or aa 10 and aa 14 , or aa 12 and aa 16 They are optionally cyclized via their side chains to form lactam bonds; and aa 10 aa 17 aa 20 , or aa 24 Assume that one or at least one of these is a natural or unnatural amino acid U covalently bonded to X.

[0160] In some embodiments, the peptide products described herein have the structure of formula III-B: His1-aa2-aa3-Gly4-Thr5-aa6-Thr7-Ser8-Asp9-aa 10 -aa 11 -aa 12 -aa 13 -aa 14 -aa 15 -aa 16 -aa 17 -aa18 -aa 19 -aa 20 -aa 21 -aa 22 -aa 23 -aa 24 -aa 25 -aa 26 -aa 27 -aa 28 -aa 29 -aa 30 -Z Form III-B (SEQ.ID.NO.3) During the ceremony: Z is OH, or -NH-R 3 And here, R 3 C1-C is H or substituted or unsubstituted. 12 Alkyl; or a PEG chain less than 10 Da; aa2 is Gly, MePro, or Aib; aa3 is either Gln or Cit; aa6 is Phe, 2FPhe, MePhe, 2FMePhe, or Nal2; aa 10 is Tyr, Nal2, Bip, Bip2EtMeO, or U(X); aa 11 It does not exist, or it is Ser, Asn, Bip, or U(X); aa 12 It does not exist, or it is Lys, Glu, Ser, or U(X); aa 13 It does not exist, or it is Tyr, Gln, Cit, or U(X); aa 14 It does not exist, or it is Leu, Nle, or U(X); aa 15 It does not exist, or it is Asp, Glu, or U(X); aa 16 It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Lys, Arg, or U(X); aa 17 It does not exist, or it is Arg, hArg, Gln, Glu, Lys, Cit, Aib, or U(X); aa 18 It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X); aa 19 It does not exist, or it is Ala, Aib, or U(X); aa 20 It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, or U(X); aa 21 It does not exist, or it is Asp, Glu, Leu, Aib, or U(X); aa 26 It does not exist, or it is Phe or U(X); aa 23 It does not exist, or it is Val, Ile, Aib, or U(X); aa 24 It does not exist, or it is Ala, Gln, or U(X); aa 25 It does not exist, or it is Trp or U(X); aa 26 It does not exist, or it is Leu or U(X); aa 27 It does not exist, or it is Met, Val, Leu, Nle, Lys, or U(X); aa 28 It does not exist, or it is Asn, Gln, Cit, or U(X); aa 29 It does not exist, or it is Thr, Aib, or U(X); aa 30 It does not exist, or it is Arg or U(X); Here, aa1-aa 23 Any two of them can be optionally cyclized via their side chains to form a lactam bond; and aa 10 aa 11 aa 12 aa 16 aa 17 aa 18 aa 19 aa20 aa 21 aa 22 aa 23 aa 24 , or aa 28 Assume that one or at least one of these is a natural or unnatural amino acid U that is covalently bonded to X.

[0161] In some specific embodiments of formulas III-A and III-B, X has the following structure:

[0162] [ka] During the ceremony: R 1a C1-C is either substituted or non-substituted. 30 It is an alkyl group; R 1b , R 1c , and R 1d is H; W 1 is -(C=O)-NH-; W 2 is -O- and R 2 It is a single bond.

[0163] In some of the embodiments described above, R 1a C1-C 20 Alkyl alkyl group, C8-C 20 Alkyl alkyl group, C 12-18 Alkyl alkyl group, or C 14 -C 18 It is an alkyl group.

[0164] In some embodiments of Formula III-B, U is any linker amino acid as described herein. Table 1 in Figure 1, Table 2 in Figure 2, and Table 3 in Figure 3 show specific examples of peptides covalently bonded with surfactants as described herein.

[0165] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-aa2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu 16 -U(X) 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -NH2;(SEQ.ID.NO.795) During the ceremony aa2 is either Gly or Aib; aa 28 It is either Asn or Gln.

[0166] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -aa 16 -Gln 17 Ala 18 -Ala 19 -aa 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega-X) 24 -Trp 25 -Leu 26 -Leu 27 -aa 28 -Thr 29 -NH2;(SEQ.ID.NO.796) During the ceremony aa 16 and aa 20 Each of these is individually either Lys or Glu, and is cyclized via their side chains to form a lactam bond; and aa 28 is either Asn or Gln; X comprises a glucuronyl class portion prepared from 1-alkyl beta-D-glucoside, 1-alkyl beta-D-maltoside, 1-alkyl beta-D-melibioside (melibiosides), or the corresponding alpha-glycoside, where the alkyl is C8-C 20 It is a linear alkyl chain.

[0167] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega-X) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.797) During the ceremony aa 16 and aa 20 They are cyclized via their side chains to form lactam bonds; and X comprises a glucuronyl class portion prepared from a 1-alkyl beta-D-glucoside, 1-alkyl beta-D-maltoside, 1-alkyl beta-D-melibiose, or the corresponding alpha-glycoside, where the alkyl is C8-C 20 It is a linear alkyl chain.

[0168] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-dodecyl beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.601) During the ceremony Glu * 16 and Lys * 20 These are cyclized via their side chains to form lactam bonds.

[0169] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-tetradecyl beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.602) During the ceremony Lys * 16 and Lys * 20 These are cyclized via their side chains to form lactam bonds.

[0170] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-hexadecyl beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27-Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.601) During the ceremony Glu * 16 and Lys * 20 These are cyclized via their side chains to form lactam bonds.

[0171] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-octadecyl beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.604) During the ceremony Glu * 16 and Lys * 20 These are cyclized via their side chains to form lactam bonds.

[0172] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10-Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega(1-Octyl Beta-D-Meriviouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.630).

[0173] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega(1-Dodecyl Beta-D-Melibiouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.631).

[0174] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-tetradecyl beta-D-meribiouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.632).

[0175] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega(1-Hexadecyl Beta-D-Melibiouronyl)) 24 -Trp 25 -Leu 26 -Leu27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.633).

[0176] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-Omega(1-Octadecy Beta-D-Melibiouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.634).

[0177] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile23 -Lys(N-Omega(1-Hexadecyl Alpha-D-Meriviouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.805).

[0178] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-tetradecyl alpha-D-meribiouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.819).

[0179] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16-Lys(N-Omega(1-Hexadecyl Alpha-D-Meriviouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2; (SEQ.ID.NO.820).

[0180] In some embodiments of formula IA, III-A, or III-B, the peptide product has the following structure:

[0181] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 Lys(N-Omega(1-Octadecyl Alpha-D-Melibiouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.821).

[0182] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-Omega(1-Dodecyl Alpha-D-Meriviouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-dodecyl beta-D-glucouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1099).

[0183] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 - Lys(N-omega(1-tetradecyl alpha-D-meribiouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23-Lys(N-omega(1-tetradecyl beta-D-glucouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1100).

[0184] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-Omega(1-Hexadecyl Alpha-D-Meriviouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-hexadecyl beta-D-glucouronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1101).

[0185] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu *16 -Lys(N-omega(1-(13-carboxy-tridecyloxy beta-D-glucouronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1102).

[0186] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-(15-carboxyl-pentadecyloxy beta-D-glucuronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1103).

[0187] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Lys(N-omega(1-(17-carboxyl-heptadecyloxy beta-D-glucuronyl)) 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Gln 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1104).

[0188] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-(13-carboxyl-tridecyloxy beta-D-glucuronyl)) 24 -Trp 25-Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1105).

[0189] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20 -Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-(15-carboxyl-pentadecyloxy beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1106).

[0190] In some embodiments of formulas IA, III-A, or III-B, the peptide product has the following structure: His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu * 16 -Gln 17 -Ala 18 -Ala 19 -Lys * 20-Glu 21 -Phe 22 -Ile 23 -Lys(N-omega(1-(17-carboxyl-heptadecyloxy beta-D-glucuronyl)) 24 -Trp 25 -Leu 26 -Leu 27 -Gln 28 -Thr 29 -NH2;(SEQ.ID.NO.1107).

[0191] Within the scope of the embodiments presented herein, peptide products of formula IA, formula III-A, or formula III-B are considered, and the peptide product comprises one or more surfactants (for example, group X having the structure of formula I). ​​In one embodiment, the peptide product of formula IA, formula III-A, or formula III-B comprises one surfactant. In another embodiment, the peptide product of formula IA, formula III-A, or formula III-B comprises two surfactants. In yet another embodiment, the peptide product of formula IA, formula III-A, or formula III-B comprises three surfactants.

[0192] This specification recognizes the importance of specific portions of SEQ.ID.NO.1 for the treatment of diseases associated with insulin resistance and / or cardiovascular diseases. Accordingly, this specification provides a method for treating diabetes in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1. 17 The process includes administering a glucagon analog containing the specified compound.

[0193] In a further embodiment, this specification provides a method for treating diabetes in an individual as needed, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 18 The process includes administering a glucagon analog containing the specified compound.

[0194] In another embodiment, this specification provides a method for treating diabetes in an individual in need, the method comprising: a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 in the individual. 19 The process includes administering a glucagon analog containing the specified compound.

[0195] In another embodiment, this specification provides a method for treating diabetes in an individual in need, the method comprising: a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 in the individual. 20 The process includes administering a glucagon analog containing the specified compound.

[0196] In an additional embodiment, the administration of the glucagon analog described above causes weight loss.

[0197] This specification recognizes the importance of specific portions of SEQ.ID.NO.1 for the treatment of diseases associated with insulin resistance and / or cardiovascular diseases. Accordingly, this specification provides a method for treating diabetes in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1. 17 The process includes administering a glucagon analog containing the specified compound.

[0198] In a further embodiment, this specification provides a method for treating diabetes in an individual as needed, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 18 The process includes administering a glucagon analog containing the specified compound.

[0199] In another embodiment, this specification provides a method for treating diabetes in an individual in need, the method comprising: a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 in the individual. 19 The process includes administering a glucagon analog containing the specified compound.

[0200] In another embodiment, this specification provides a method for treating diabetes in an individual in need, the method comprising: a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 in the individual. 20 The process includes administering a glucagon analog containing the specified compound.

[0201] In an additional embodiment, the administration of the glucagon analog described above causes weight loss.

[0202] In any of the embodiments described above, the glucagon analog is modified with the surfactant X of formula I:

[0203] [ka] During the ceremony: R 1a Independently, at each occurrence, there is a single bond, H, saccharide, substituted or unsubstituted C1-C 30 It is an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid nucleus-containing moiety; R 1b , R 1c , and R 1d Each of these independently, at each occurrence, consists of a single bond, H, and a substituted or unsubstituted C1-C bond. 30 It is an alkyl group, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group; W 1 Independently, at each occurrence, -CH2-, -CH2-O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH2-S-; W 2 is -O-, -CH2-, or -S-; R 2 Independently, at each occurrence, U is a single bond, H, substitution, or unsubstituted C1-C. 30 Alkyl groups, substituted or unsubstituted alkoxyaryl groups, substituted or unsubstituted aralkyl groups, -NH-, -S-, -triazolo-, -NH(C=O)-CH2-, -(CH2) m-It is maleimide; n is 1, 2, or 3; and m is an integer between 1 and 10.

[0204] In a specific embodiment, the glucagon analog is modified with a surfactant X having the following structure:

[0205] [ka] During the ceremony: R 1a C1-C is either substituted or non-substituted. 30 It is an alkyl group; R 1b , R 1c , and R 1d is H; W 1 is -(C=O)-NH-; W 2 is -O- and R 2 It is a single bond.

[0206] In some of the embodiments described above, R 1a C1-C 20 Alkyl alkyl group, C8-C 20 Alkyl alkyl group, C 12 -C 18 Alkyl alkyl group, or C 14 -C 18 It is an alkyl group.

[0207] In some embodiments, R 2 Independently, at each occurrence, R is a single bond, H, leaving group, protecting group, or a reversibly protected natural or unnatural amino acid. In some embodiments, R 2 In some embodiments, R is a single bond, independently of the others at each occurrence. 2 Independently, at each occurrence, is H. In some embodiments, R 2 Independently, each instance is a leaving group. In some embodiments, R 2 Independently, each instance is a protecting group. In some embodiments, R2 R is independently a reversibly protected natural or non-natural amino acid at each occurrence. In some embodiments, R 2 These are independent of each other, and at each occurrence, C1-C is either substituted or unsubstituted. 30 It is an alkyl group. In some embodiments, R 2 R is independently a substituted or unsubstituted alkoxyaryl group at each occurrence. In some embodiments, R 2 Independently, each occurrence is a substituted or unsubstituted aralkyl group. In some embodiments, R 2 Independently, at each occurrence, it is -NH-. In some embodiments, R 2 Independently, at each occurrence, is -S-. In some embodiments, R 2 Independently, at each occurrence, is -triazolo-. In some embodiments, R 2 Independently, at each occurrence, it is -NH-(C=O)-CH2-. In some embodiments, R 2 These are independent, and each occurrence is -(CH2) m It is maleimide.

[0208] In the foregoing and in some embodiments herein, R 1a is a saccharide. In some embodiments, the saccharide is galactose. In certain embodiments, the saccharide is alpha-linked galactose. In other embodiments, the saccharide is alpha-linked galactopyranose, beta-linked galactopyranose, alpha-linked galacofuranose, or beta-linked galacofuranose.

[0209] As used herein, the term diabetes mellitus includes both type I and type II diabetes mellitus. Accordingly, in some embodiments, the methods described herein involve administering any of the compounds described herein to an individual suffering from type I diabetes mellitus, wherein the compounds include compounds of formula II, III-A, and / or III-B, and / or compounds listed in Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3. In some other embodiments, the methods described herein involve administering any of the compounds described herein to an individual suffering from type II diabetes mellitus, wherein the compounds include compounds of formula II, III-A, and / or III-B, and / or compounds listed in Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3.

[0210] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 17 The process includes administering a glucagon analog containing the specified compound.

[0211] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 18 The process includes administering a glucagon analog containing the specified compound.

[0212] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 19 The process includes administering a glucagon analog containing the specified compound.

[0213] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 20 The process includes administering a glucagon analog containing the specified compound.

[0214] In the embodiments described above, if the cardiovascular disease is related to an ischemic event, the glucagon analog may be administered.

[0215] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 17 The process includes administering a glucagon analog containing the specified compound.

[0216] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 18 The process includes administering a glucagon analog containing the specified compound.

[0217] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 19 The process includes administering a glucagon analog containing the specified compound.

[0218] This specification also provides a method for treating cardiovascular disease in an individual in need, the method comprising administering to the individual a therapeutically effective amount of amino acid residues aa1-aa of SEQ.ID.NO.1 20 The process includes administering a glucagon analog containing the specified compound.

[0219] In the embodiments described above, if the cardiovascular disease is related to an ischemic event, the glucagon analog may be administered.

[0220] In any of the embodiments described above, the glucagon analog is modified with the surfactant X of formula I:

[0221] [ka] During the ceremony: R 1aIndependently, at each occurrence, there is a single bond, H, saccharide, substituted or unsubstituted C1-C 30 It is an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid nucleus-containing moiety; R 1b , R 1c , and R 1d Each of these independently, at each occurrence, consists of a single bond, H, and a substituted or unsubstituted C1-C bond. 30 It is an alkyl group, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group; W 1 Independently, at each occurrence, -CH2-, -CH2-O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH2-S-; W 2 is -O-, -CH2-, or -S-; R 2 Independently, at each occurrence, in each occurrence, a single bond to U, H, substituted or unsubstituted C1-C 30 Alkyl groups, substituted or unsubstituted alkoxyaryl groups, substituted or unsubstituted aralkyl groups, -NH-, -S-, -triazolo-, -NH(C=O)-CH2-, -(CH2) m -It is maleimide; n is 1, 2, or 3; and m is 1-10.

[0222] In a specific embodiment, the glucagon analog is modified with a surfactant X having the following structure:

[0223] [ka] During the ceremony: R 1a C1-C is either substituted or non-substituted. 30 It is an alkyl group; R 1b , R 1c , and R 1d is H; W 1 is -(C=O)-NH-; W 2 is -O- and R 2 It is a single bond.

[0224] In some of the embodiments described above, R 1a C1-C 20 Alkyl alkyl group, C8-C 20 Alkyl alkyl group, C 12 -C 18 Alkyl alkyl group, or C 14 -C 18 It is an alkyl group.

[0225] In the foregoing and in some embodiments herein, R 1a is a saccharide. In some embodiments, the saccharide is galactose. In certain embodiments, the saccharide is alpha-linked galactose. In other embodiments, the saccharide is alpha-linked galactopyranose, beta-linked galactopyranose, alpha-linked galacofuranose, or beta-linked galacofuranose.

[0226] Modifications at the amino or carboxyl terminus may be optionally introduced into peptides (e.g., glucagon or GLP-1) (Nestor, JJ, Jr. (2009) Current Medicinal Chemistry 16:4399-4418). For example, peptides can be shortened or acylated at the N-terminus to produce low-efficacy peptide analogs, partial agonists, and antagonists, as seen in some peptides (Gourlet, P., et al. (1998) Eur J Pharmacol 354:105-111, Gozes, I. and Furman, S. (2003) Curr Pharm Des 9:483-494), the details of which are incorporated herein by reference). For example, deletion of the first six residues of bPTH results in an antagonistic analog (Mahaffey, JE, et al. (1979) J Biol Chem 254:6496-6498; Goldman, ME, et al. (1988) Endocrinology 123:2597-2599), and similar manipulations of the peptides described herein generate potent antagonistic analogs. Other modifications to the N-terminal group of the peptide, such as deletion or incorporation of D-amino acids like D-Phe, can also yield potent and long-acting agonists or antagonists when substituted with modifications described herein, such as long-chain alkyl glycosides. Such agonists and antagonists also have commercial utility and fall within the scope of the thoughtful embodiments described herein.

[0227] Within the scope of the embodiments described herein, surfactants covalently bonded to peptide analogs are also considered, and natural peptides are modified by acetylation, acylation, pegylation, ADP-ribosylation, amidation, covalent bonding of lipids or lipid derivatives, covalent bonding of phosphatidylinositol, crosslinking, cyclization, disulfide bond formation, demethylation, formation of covalent cysteine ​​crosslinks, pyroglutamate formation, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodization, methylation, myristoylation, oxidation, proteolysis, phosphorylation, prenylation, racemization, glycosylation, lipid bonding, sulfation, gamma-carboxylation, hydroxylation and ADP-ribosylation of glutamic acid residues, selenylation, sulfation, arginylation, and RNA-mediated addition of amino acids to proteins, as well as ubiquitination. For example, (Nestor, JJ, Jr. (2007) Comprehensive Medicinal Chemistry II 2:573-601, Nestor, JJ, Jr. (2009) Current Medicinal Chemistry 16:4399-4418, Creighton, TE (1993), Wold, F. (1983) Posttranslational Covalent Modification of Proteins 1-12, Seifter, S. and Englard, S. (1990) Methods Enzymol 182:626-646, Rattan, SI, et al. (1992) Ann NY Acad Sci See 663:48-62). Within the scope of the embodiments described herein, branched or cyclic peptides, with or without branching, are also considered. Cyclic branched peptides and branched cyclic peptides may result from post-translational natural processes or be produced by appropriate synthetic methods. In some embodiments, any peptide product described herein includes the above-mentioned peptide analogs, which are later covalently bonded to an alkyl-glycoside surfactant moiety.

[0228] Within the scope of the embodiments presented herein, for example, peptide chains having fatty acids such as octane, decane, dodecane, tetradecane, hexadecane, octadecane, and 3-phenylpropanoic acid, and saturated or unsaturated alkyl chains, with linker amino acids appropriately substituted at the ε-position of Lys (Zhang, L. and Bulaj, G. (2012) Curr Med Chem 19:1602-1618). Similarly, such acylations may be bound to a spacer such as gamma-linked glutamic acid, or the gamma-linked glutamic acid may be further bound to a “mini-PEG” chain such as 9-amino-4,7-dioxanonoic acid (DiMarchi, R. and Ward, BP (2012) US Patent Application US2012 / 0238493). Exemplary examples of such analogs are listed below, but are not limited to:

[0229] His1-Gly2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu 16 -Lys(N-epsilon-dodecanoyl) 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22 -Ile 23 -Ala 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.785)

[0230] His1-Gly2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr13 -Leu 14 -Asp 15 -Glu 16 -Lys(N-epsilon-hexadecanoyl) 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22 -Ile 23 -Ala 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.786)

[0231] His1-Gly2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Glu 16 -Lys(N-epsilon-octadecanoyl) 17 -Ala 18 -Ala 19 -Lys 20 -Glu 21 -Phe 22 -Ile 23 -Ala 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.787)

[0232] His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Cyclo(Glu 16 -Gln17 -Ala 18 -Ala 19 -Lys 20 )-Glu 21 -Phe 22 -Ile 23 -Lys(N-epsilon-hexadecanoyl) 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.788)

[0233] His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Cyclo(Glu 16 -Gln 17 -Ala 18 -Ala 19 -Lys 20 )-Glu 21 -Phe 22 -Ile 23 -Lys(N-epsilon(N-alpha-octadecanoyl)-gamma-glutamyl) 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.789)

[0234] His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Cyclo(Glu 16 -Gln 17 -Ala 18 -Ala 19 -Lys20 )-Glu 21 -Phe 22 -Ile 23 -Lys(N-epsilon(N-alpha-hexadecanoyl)-gamma-glutamyl) 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2(SEQ.ID.NO.790)

[0235] His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Cyclo(Glu 16 -Gln 17 -Ala 18 -Ala 19 -Lys 20 )-Glu 21 -Phe 22 -Ile 23 -Lys(N-epsilon-(N-alpha-octadecanoyl(N9-gamma-glutamyl(9-amino-4,7-dioxanonanyl)))) 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.791)

[0236] His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Cyclo(Glu 16 -Gln 17 -Ala 18 -Ala 19 -Lys20 )-Glu 21 -Phe 22 -Ile 23 -Lys(N-epsilon(N-alpha-hexadecanoyl(N9-gamma-glutamyl(9-amino-4,7-dioxanonanyl)))) 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.792), etc.

[0237] In further embodiments, the peptide chain is optionally substituted at appropriate positions by reaction of a linker amino acid (e.g., sulfhydryl of Cys) with a spacer and hydrophobic moiety, such as a steroid nucleus (e.g., a cholesterol moiety). In some such embodiments, the modified peptide further comprises one or more PEG chains. Exemplary examples of such molecules are, but are not limited to, the following:

[0238] His1-Aib2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Cyclo(Glu 16 -Gln 17 -Ala 18 -Ala 19 -Lys 20 )-Glu 21 -Phe 22 -Ile 23 -Cys(S-(3-(PEG4-aminoethylacetamide-cholesterol)) 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.793)

[0239] His1-Gly2-Gln3-Gly4-Thr5-Phe6-Thr7-Ser8-Asp9-Tyr 10 -Ser 11 -Lys 12 -Tyr 13 -Leu 14 -Asp 15 -Cyclo(Glu 16 -Gln 17 -Ala 18 -Ala 19 -Lys 20 )-Glu 21 -Phe 22 -Ile 23 -Cys(S-(3-(PEG4-aminoethylacetamide-cholesterol)) 24 -Trp 25 -Leu 26 -Leu 27 -Asn 28 -Thr 29 -NH2, (SEQ.ID.NO.794), etc.

[0240] In addition to the 20 standard amino acids, there are a vast number of "non-standard amino acids," or unnatural amino acids known in the art and incorporated into the compounds described herein as described above. Other non-standard amino acids are modified with side chains reactive to conjugation (Gauthier, MA and Klok, HA (2008) Chem Commun (Camb) 2591-2611; de Graaf, AJ, et al. (2009) Bioconjug Chem 20:1281-1295). In one method, the developed tRNA / tRNA synthetase pair is encoded in the expression plasmid by an amber suppressor codon (Deiters, A, et al. (2004). Bio-org. Med. Chem. Lett. 14, 5743-5). For example, p-azidophenylalanine was incorporated into a peptide and reacted with a functionalized surfactant or a PEG polymer having an acetylene moiety in the presence of a reducing agent and copper ions to facilitate an organic reaction known as "Huisgen[3+2] cycloaddition." Similar reaction sequences using the reagents described herein, comprising acetylene-modified alkyl glycosides or PEG-modified glycosides, result in PEGylated or alkyl glycoside-modified peptides. For peptides with fewer than approximately 50 residues, standard solid-phase synthesis is used for the incorporation of the reactive amino acid residues at desired positions in the chain. Such surfactant-modified peptides and / or proteins yield a spectrum of pharmacological and medical properties distinct from peptides modified solely by PEG incorporation.

[0241] Those skilled in the art will recognize that numerous permutations of peptide analogs are possible, and if they have a surfactant portion incorporating an amino acid sequence, they will possess the desired properties of peptides modified with the surfactants described herein.

[0242] <Specific Definition> As used herein, “a” or “an” means one or more. As used in the claims, when used with the word “comprising,” the word “a” or “an” means one or more. As used herein, “another” means at least two or more.

[0243] As used herein, one- and three-letter abbreviations for various common amino acids are as recommended in Pure Appl. Chem. 31, 639-645 (1972) and 40, 277-290 (1974), and in accordance with 37 CFR § 1.822 (55 FR 18245, May 1, 1990). These abbreviations represent L-amino acids unless otherwise explicitly stated as D- or DL. Certain amino acids, both natural and unnatural, are achiral, such as glycine, Cα-diethylglycine (Deg), α-aminoisobutyric acid (Aib), 1-aminocyclobutane-1-carboxylic acid (Ac4c), 1-aminocyclopentane-1-carboxylic acid (Ac5c), and 1-aminocyclohexane-1-carboxylic acid (Ac6c). Analogues of glutamine include citrulline (Cit). All peptide sequences are provided with the left-side N-terminal amino acid and the right-side C-terminal amino acid. C-alpha-methylproline (MePro) may be used to inhibit peptide bonds, such as C-alpha-methylphenylalanine (MePhe), 2-fluorophenylalanine (2FPhe), C-alpha-methyl-2-fluorophenylalanine (2FMePhe), and C-alpha-methyllysine (MeLys). Additional non-natural aromatic amino acids, such as 2-naphthylalanine (Nal2), biphenylalanine (Bip), and 2-ethyl-4'-methoxybiphenylalanine (Bip2EtMeO), may be substituted to increase efficacy.

[0244] The “alkyl” group refers to an aliphatic hydrocarbon group. References to alkyl groups include “saturated alkyl” and / or “unsaturated alkyl.” Alkyl groups, whether saturated or unsaturated, include branched, linear, or cyclic groups. A “substituted” alkyl group is substituted with one or more additional groups. In certain embodiments, one or more additional groups are individually and independently selected from amides, esters, alkyls, cycloalkyls, heteroalkyls, aryls, heteroaryls, heteroalicyclics, hydroxys, alkoxys, alkylthios, arylthios, alkyl sulfoxides, aryl sulfoxides, esters, alkylsulfones, arylsulfones, cyanos, halogens, alcoyls, alcoyloxos, isocyanates, thiocyanates, isothiocyanates, nitros, haloalkyls, haloalkoxys, fluoroalkyls, aminos, alkyl-aminos, dialkyl-aminos, amides, oxos, hydrophobic natural products such as steroids, aralkyl chains (including alkoxyaryls), alkyl chains containing acyl moieties, and the like. In some embodiments, alkyl groups are attached to the Nα-position of a residue in the peptide (e.g., Tyr or Dmt). This class is referred to as N-alkyl, and C1-C 10 The alkyl group comprises a linear or branched alkyl group, or an aryl-substituted alkyl group such as benzyl or phenylethyl. In some embodiments, the alkyl group is a 1-alkyl group in a glycosidic bond to the saccharide group (typically, for example, at one position of glucose). Such a 1-alkyl group is C1-C 30 It is an alkyl group.

[0245] The "aryl" group represents an aromatic ring in which each of the ring-forming atoms is a carbon atom. The aryl rings described herein include rings having 5, 6, 7, 8, 9, or more than 9 carbon atoms. The aryl group may be optionally substituted with substituents selected from halogens, alkyls, acyls, alkoxys, alkylthios, sulfonyls, dialkyl-aminos, carboxylates, cyanos, and others. Examples of aryl groups include, but are not limited to, phenyls and naphthalenyls.

[0246] The term "acyl group" is C1-C 20 This refers to an acyl chain. This chain may include linear aliphatic chains, branched aliphatic chains, chains containing cyclic alkyl moieties, hydrophobic natural products such as steroids, aralkyl chains, or alkyl chains containing acyl moieties.

[0247] The term "steroid nucleus" refers to the core of a steroid, including the arrangement of four fused rings, explicitly labeled A, B, C, and D, as shown below:

[0248] [ka] Examples of steroid nucleus-containing regions include, but are not limited to, cholesterol.

[0249] As used herein, “therapeutic composition” may comprise a mixture with an aqueous or organic carrier or excipient, and may be mixed with a common, non-toxic, and pharmaceutically acceptable carrier for, for example, tablets, pellets, capsules, lyophilized bodies, suppositories, solutions, emulsions, suspensions, or other forms suitable for use. In addition to those disclosed above, carriers may include, in solid, semi-solid, or liquid form, arginate, collagen, glucose, lactose, mannose, acacia gum, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea, medium-chain triglycerides, dextran, and other carriers suitable for use in the preparation. In addition, auxiliary stabilizers, thickeners, or colorants may be used, such as desiccants such as triurose.

[0250] As used herein, “pharmaceutically acceptable carrier” or “therapeutably effective carrier” may be aqueous or non-aqueous (solid), alcoholic or oily, or a combination thereof, and may include surfactants, mitigants, lubricants, stabilizers, dyes, fragrances, preservatives, acids or bases for pH adjustment, solvents, emulsifiers, gelling agents, humectants, stabilizers, wetting agents, time-release agents, water-retaining agents, or other components commonly found in certain forms of pharmaceutical compositions. Pharmaceutically acceptable carriers are well known in the art and include, for example, water or physiologically buffered saline, or aqueous solutions of other solvents or vehicles such as glycols or glycerol, and oils such as olive oil or injectable organic esters. A pharmaceutically acceptable carrier may include a physiologically acceptable compound that acts to stabilize or increase the absorption of, for example, specific inhibitors, hydrocarbons such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, other stabilizers, or excipients.

[0251] As used herein, the “insulin-resensitizing” amount of a peptide product is the amount that increases the body’s response to endogenously or exogenously administered insulin in an individual in need, as demonstrated by, for example, an oral glucose-triggered test or a clamp test for normal blood glucose, while typically causing a decrease in body weight.

[0252] The pharmaceutical composition also includes other pharmaceutically acceptable auxiliary substances that are required to approximate physiological conditions, and such “substances” include, but are not limited to, pH adjusters and buffers, isotonic agents, and for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, etc. In addition, the peptide, or its modification or suspension, may contain lipid protectants that protect lipids from damage by free radicals and lipid peroxidation after storage. Lipophilic free radical quenchers such as α-tocopherol and water-soluble iron-specific chelating agents such as ferrioxamine are suitable.

[0253] As used herein, “surfactant” is a surfactant that modifies the surface tension of water. Typically, a surfactant has one lipophilic and one hydrophilic group or region in its molecule. Broadly speaking, the groups include a variety of soaps, detergents, emulsifiers, dispersants, and wetting agents, and preservatives. More specifically, surfactants include stearyltriethanolamine, sodium lauryl sulfate, sodium taurocholate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glyceryl monostearate; and hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, polyethylene glycol (PEG), sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, or alkyl glycosides. In some embodiments, the surfactant is a nonionic surfactant (e.g., alkyl glycoside surfactant). In some embodiments, the surfactant is an ionic surfactant.

[0254] As used herein, “alkyl glycoside” refers to any sugar linked by linkage with any hydrophobic alkyl, as is known in the art. The hydrophobic alkyl can be selected for any desired size, depending on the desired hydrophobicity and hydrophilicity of the saccharide moiety. In one aspect, the alkyl chain ranges from 1 to 30 carbon atoms; or 6 to 16 carbon atoms.

[0255] As used herein, “saccharide” includes monosaccharides, oligosaccharides, or polysaccharides in a linear or ring-like structure. Oligosaccharides are saccharides having two or more monosaccharide residues. Some examples of the many possible saccharides suitable for use in functionalized forms include glucose, galactose, maltose, maltotriose, maltotetraose, sucrose, and trehalose.

[0256] As used herein, “sucrose ester” refers to the sucrose ester of a fatty acid. Sucrose esters can take many forms, from larger, bulkier lipid acetates reacted with sucrose, to the eight hydroxyl groups in sucrose available for reaction, and many fatty acid groups. This flexibility means that many products and functionalities can be adjusted based on the fatty acid portion used. Sucrose esters have food and non-food applications, particularly as surfactants and emulsifiers, and are increasingly used in pharmaceuticals, cosmetics, detergents, and feed additives. They are biodegradable, non-toxic, and skin-friendly.

[0257] As used herein, “suitable” alkyl glycoside means one that is non-toxic and non-ionic. In some cases, suitable alkyl glycosides, when administered with a compound via the eye, nose, nasolacrimal duct, sublingual gland, buccal, inhalation route, or by infusion route such as subcutaneous, intramuscular, or intravenous route, reduce immunogenicity or aggregation and increase the bioavailability of the compound.

[0258] A "linker amino acid" is any natural or non-natural amino acid that contains a reactive functional group used for covalent bonding with a functionalized surfactant (de Graaf, AJ, et al. (2009) Bioconjug Chem 20:1281-1295). For example, in some embodiments, the linker amino acid is Lys or Orn with a reactive functional group -NH2; or Cys with a reactive functional group -SH; or Asp or Glu with a reactive functional group -C(=O)-OH. For example, in some other embodiments, the linker amino acid is any amino acid that has a reactive functional group such as -OH, -N3, haloacetyl, or acetylene group, which is used for covalent bonding with a appropriately functionalized surfactant.

[0259] As used herein, “functionalized surfactant” is a surfactant comprising a reactive group suitable for covalent bonding with linker amino acids. For example, in some embodiments, the functionalized surfactant comprises a carboxylic acid group (e.g., at the 6-position of a monosaccharide) as a reactive group suitable for covalent bonding with linker amino acids. For example, in some embodiments, the functionalized surfactant comprises an -NH2 group, -N3 group, acetylene group, haloacetyl group, -O-NH2 group, or -(CH2-)m-maleimide group (e.g., at the 6-position of a monosaccharide, as shown in schematic Figure 6) that enables covalent bonding with suitable linker amino acids. In some embodiments, the functionalized surfactant is a compound of formula II as described herein. Optionally, in some specific embodiments, the functionalized surfactant comprises a covalently bonded linker amino acid; the peptide to be modified with the surfactant is then formed by the sequential addition of one or more amino acids to the linker amino acid.

[0260] As used herein, the term “peptide” refers to any peptide containing two or more amino acids. The term peptide includes polypeptides, short peptides (e.g., peptides containing 2 to 14 amino acids), intermediate-length peptides (15 to 50), or long-chain peptides (e.g., proteins). The terms peptide, polypeptide, intermediate-length peptide, and protein may be used interchangeably herein. As used herein, the term “peptide” is interpreted to mean a polymer composed of amino acid residues, associated spontaneously occurring structural variants, and unspontaneous analogs of its synthesis, linked via peptide bonds, associated spontaneously occurring structural variants, and unspontaneous analogs of its synthesis. Synthetic peptides may be synthesized, for example, using an automated peptide synthesizer.

[0261] Peptides may contain amino acids other than those encoded by 20 genes. “Peptides” include those modified by natural processes such as processing and other post-translational modifications, or by chemical modification techniques. Such modifications are well described in basic texts and in more detailed monographs and are well known to those skilled in the art. In some embodiments, it is recognized that the same type of modification is present to the same or different degrees at various sites in a given peptide. Furthermore, a given peptide may, in some embodiments, encompass more than one type of modification. Modifications occur anywhere in the peptide, including the peptide backbone, amino acid side chains, and amino or carboxyl termini.

[0262] The term peptide includes peptides or proteins, which include natural and non-natural amino acids or analogs of natural amino acids. As used herein, "analogs" of peptides and / or proteins include non-natural amino acids based on individual amino acids, such as tyrosine analogs, including para-substituted tyrosine, ortho-substituted tyrosine, and meta-substituted tyrosine, where substituents on tyrosine are acetyl, benzoyl, amino, hydrazine, hydroxyamine, thiol, carboxyl, methyl, isopropyl, C2-C 20 This includes linear or branched hydrocarbons, saturated or unsaturated hydrocarbons, O-methyl groups, polyether groups, halogens, nitro groups, etc. Examples of tyr analogs include 2,4-dimethyl-tyrosine (Dmt), 2,4-diethyl-tyrosine, O-4-allyl-tyrosine, 4-propyl-tyrosine, and Cα-methyl-tyrosine. Examples of lysine analogs include ornithine (Orn), homo-lysine, and Cα-methyl-lysine (CMeLys). Examples of phenylalanine analogs include, but are not limited to, meta-substituted phenylalanine, where the substituent is a methoxy group, C1-C 20This includes alkyl groups, such as methyl groups, allyl groups, and acetyl groups. Specific examples, though not limited to them, include 2,4,6-trimethyl-L-phenylalanine (Tmt), O-methyl-tyrosine, 3-(2-naphthyl)alanine (Nal(2)), 3-(1-naphthyl)alanine (Nal(1)), 3-methyl-phenylalanine, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Tic), fluorine-treated phenylalanine, isopropyl-phenylalanine, p-azido-phenylalanine, p-acyl-phenylalanine, p-benzoyl-phenylalanine, p-iodo-phenylalanine, p-bromophenylalanine, p-amino-phenylalanine, and isopropyl-phenylalanine. Other non-standard or unnatural amino acids used in the design of peptide analogs include, but are not limited to, C-alpha-nisubstituted amino acids such as Aib, Cα-diethylglycine (Deg), and aminocyclopentane-1-carboxylic acid (Ac5c). Such amino acids frequently lead to repressed structures that are frequently biased toward the alpha helix structure (Kaul, R. and Balaram, P. (1999) Bioorg Med Chem 7:105-117). Additional examples of such unnatural amino acids useful in analog design include homo-arginine (Har). Substitution of a reduced amide bond in certain cases leads to improved protection against enzymatic degradation or alters receptor binding. For example, the incorporation of the Tic-Phe dipeptide (designed as Tic-Ψ[CH2-NH]-Ψ-Phe), in which the amide bond between residues is reduced, reduces enzymatic degradation. Accordingly, within the scope of the embodiments described herein, surfactants covalently bonded to peptides, including the modified amino acids and / or peptide analogs described above, are also considered. Specific non-natural amino acids are shown below.

[0263] [ka]

[0264] As used herein, the term “mutant” is interpreted to mean a peptide that differs from a reference peptide but retains essential properties. Typical peptide mutants differ from another reference peptide in their amino acid sequence. Generally, the differences are limited, so that the sequences of the reference peptide and the mutant are quite similar overall and identical in many regions. Mutants and reference peptides may differ in their amino acid sequence due to one or more substitutions, additions, or deletions in any combination. The substituted or inserted amino acid residues may or may not be encoded by the genetic code. Non-spontaneous mutants of peptides may be produced by mutagenesis, direct synthesis, and other appropriate recombination methods.

[0265] <Method> This specification provides, in several embodiments, methods for preventing and / or treating diseases associated with reduced insulin sensitivity, the methods comprising administering to an individual in need a therapeutically effective amount of a peptide and / or protein product modified with a surfactant described herein (e.g., a peptide product of formula IA, III-A, or III-B). In some embodiments, diseases characterized by reduced insulin sensitivity include, but are not limited to, metabolic syndrome, obesity-related insulin resistance, hypertension, systemic inflammation associated with high C-reactive proteins, and diabetes.

[0266] This specification also provides a method for treating insulin resistance, which comprises administering to an individual in need a peptide and / or protein product (e.g., a peptide product of formula IA, III-A, or III-B) modified with a surfactant described herein. In some embodiments, insulin resistance is associated with metabolic syndrome (syndrome X) and / or diabetes.

[0267] This specification also provides a method for stimulating the body's resensitization to insulin, the method comprising administering to an individual in need a peptide and / or protein product modified with a surfactant described herein (e.g., a peptide product of formula IA, III-A, or III-B).

[0268] In further embodiments, this specification also provides a method for increasing insulin sensitivity through weight loss, the method comprising administering to an individual in need a peptide and / or protein product modified with a surfactant described herein (e.g., formula IA, III-A, or III-B, and the peptide products of Table 1 in Figure 1, Table 2 in Figure 2, and Table 3 in Figure 3).

[0269] This specification also provides a method for treating diabetes or prediabetes, the method comprising administering to an individual in need a therapeutically effective amount of the peptide products described above and herein, as well as Table 1 in Figure 1, Table 2 in Figure 2, and Table 3 in Figure 3.

[0270] This specification provides a method for treating or delaying the progression or onset of a disease selected from diabetes mellitus, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, insulin resistance, hyperglycemia, hyperinsulinemia, metabolic syndrome, diabetic complications, high blood concentrations of free fatty acids or glycerol, hyperlipidemia, obesity, hypertriglyceridemia, atherosclerosis, acute cardiovascular syndrome, infarction, ischemia-reperfusion, and hypertension, the method comprising administering to the individual in need a therapeutically effective amount of the peptide products described herein and Table 1 in Figure 1, Table 2 in Figure 2, or Table 3 in Figure 3. In an additional embodiment, this specification provides a method for treating delayed wound healing, the method comprising administering to the individual in need a therapeutically effective amount of the peptide products described herein and Table 1 in Figure 1, Table 2 in Figure 2, and Table 3 in Figure 3.

[0271] In one embodiment, the disease being treated is diabetes mellitus. In one embodiment, the disease being treated is insulin resistance. In one embodiment, the disease being treated is metabolic syndrome. In one embodiment, the effective amount of the peptide is about 0.1 μg / kg / day to about 100.0 μg / kg / day.

[0272] In one embodiment, the method of administration is parenteral. In one embodiment, the method of administration is oral. In one embodiment, the method of administration is subcutaneous. In one embodiment, the method of administration is nasal inhalation.

[0273] This specification further provides a method for reducing or inducing weight gain, the method comprising administering to the individual in question a therapeutically effective amount of the peptide products of this specification and Table 1 in Figure 1, Table 2 in Figure 2, and Table 3 in Figure 3. In some embodiments, weight gain is associated with metabolic syndrome.

[0274] This specification provides a method for treating hypoglycemia, the method comprising administering to an individual in need a therapeutically effective amount of the peptide products specified herein and those listed in Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3.

[0275] This specification also provides a method for treating diabetes mellitus, the method comprising administering to an individual in need a therapeutically effective amount of the peptide products of this specification and Table 1 of Figure 1, Table 2 of Figure 2, and Table 3 of Figure 3, and at least one additional therapeutic agent, wherein the therapeutic agent is selected from antidiabetic agents, anti-obesity agents, satiety agents, anti-inflammatory agents, anti-vasopressors, anti-atherosclerotic agents, and anti-hyperlipidemia agents.

[0276] In some embodiments of the methods described above, the peptide and / or protein covalently bonded to the surfactant is glucagon or GLP-1 peptide, or an analog thereof. In some embodiments, the surfactant-modified peptide and / or protein (e.g., peptide products of formula IA, III-A, or III-B) is administered prophylactically to delay the onset of any disease associated with insulin resistance, including, but not limited to, metabolic syndrome, hypertension, diabetes mellitus, type 2 diabetes mellitus, gestational diabetes mellitus, hyperlipidemia, atherosclerosis, and systemic inflammation. In some embodiments, the surfactant-modified peptide and / or protein (e.g., peptide products of formula IA, III-A, or III-B) is administered therapeutically to delay the progression of any disease associated with metabolic syndrome, hypertension, diabetes mellitus, type 2 diabetes mellitus, gestational diabetes mellitus, hyperlipidemia, atherosclerosis, and systemic inflammation. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of formula IA, III-A, or III-B) are administered prophylactically and / or therapeutically to slow the progression of insulin resistance in diabetes. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of formula IA, III-A, or III-B) are administered prophylactically and / or therapeutically to reduce or halt further loss of insulin resistance, thereby stabilizing the disease.

[0277] In some embodiments, the surfactant-modified peptides and / or proteins (e.g., peptide products of formula IA, III-A, or III-B) are administered parenterally. In some embodiments, the surfactant-modified peptides and / or proteins (e.g., peptide products of formula IA, III-A, or III-B) are administered subcutaneously. In some embodiments, the surfactant-modified peptides and / or proteins (e.g., peptide products of formula IA, III-A, or III-B) are administered by nasal inhalation.

[0278] In some embodiments of the methods described above, surfactant-modified peptides and / or proteins (e.g., peptide products of formula IA, III-A, or III-B) have a longer duration of action compared to pharmaceuticals including currently known therapeutic agents (e.g., exenatide, metformin, etc.).

[0279] <Combination therapy> In some embodiments of the methods described above, surfactant-modified peptides and / or proteins (e.g., peptide products of formulas IA, III-A, or III-B) are administered in combination with other treatments for metabolic syndrome, selected from the group including antidiabetic drugs, anti-obesity agents, anti-vasopressor drugs, anti-atherosclerotic agents, and anti-hyperlipidemia agents. As an example, effective antidiabetic drugs suitable for administration in combination with the surfactant-modified peptides and / or protein products described herein include biguanides, sulfonylureas, glucosidase inhibitors, PPARγ agonists, PPARα / γ dual agonists, aP2 inhibitors, DPP4 inhibitors, insulin resistance improvers, GLP-1 analogs, insulin, and meglitinide. Additional examples include metformin, glibride, glimepiride, glipyride, glipizide, chlorpropamide, gliclazide, acarbose, miglitol, pioglitazone, troglitazone, rosiglitazone, muraglitazar, insulin, Gl-262570, isaglitazone, JTT-501, NN-2344, L895 645, YM-440, R-119702, A19677, repaglinide, nateglinide, KAD1129, AR-HO 39242, GW-40 I 5 44, KRP2 I 7, AC2993, LY3 I 5902, NVP-DPP-728A, and saxagliptin.

[0280] In some embodiments of the methods described above, surfactant-modified peptides and / or proteins (e.g., peptide products of formulas IA, III-A, or III-B) are administered in combination with other treatments for metabolic syndrome, selected from a group of effective anti-obesity agents. Examples of effective anti-obesity agents suitable for administration with the peptide products described herein include beta-3 adrenergic agonists, lipase inhibitors, serotonin (and dopamine) reuptake inhibitors, thyroid receptor beta compounds, CB-1 antagonists, NPY-Y2 and NPY-Y4 receptor agonists, and appetite suppressants. Specific members of such a class include orlistat, AfL-962, A1967l, L750355, CP331648, sibutramine, topiramate, axokine, dextroamphetamine, phentermine, phenylpropanolamine, limonaban (SR1 4I7164), and mazindol.

[0281] In some embodiments of the methods described above, surfactant-modified peptides and / or proteins (e.g., peptide products of formulas IA, III-A, or III-B) are administered in combination with other treatments for metabolic syndrome, selected from a group of effective antihyperlipidemic agents. For example, effective antihyperlipidemic agents suitable for administration with the peptide products described herein include agents selected from the group consisting of MTP inhibitors, cholesterol ester transfer proteins, HMG-CoA reductase inhibitors, squalene synthetase inhibitors, fibrinate derivatives, LDL receptor activity upregulators, lipoxygenase inhibitors, and ACAT inhibitors. Specific examples of such a class include pravastatin, lovastatin, simvastatin, atorvastatin, cerivastatin, fluvastatin, nisvastatin, bisastatin, fenofibrate, gemfibrozil, clofibrate, abasimib, TS-962, MD-700, CP-52941 4, and LY295 427.

[0282] In some embodiments of the methods described above, surfactant-modified peptides and / or proteins (e.g., peptide products of formulas IA, III-A, or III-B) are administered in combination with peptide hormones and their analogues known to exhibit pro-satiety effects in animal models and humans. Within the scope of the embodiments presented herein, combinations of the peptide products described herein with long-acting satiety agents for the treatment of obesity are considered. Examples of such peptide satiety agents include GLP-1 pancreatic polypeptide (PP), cholecystokinin (CCK), peptide YY (PYY), amylin, calcitonin, OXM, neuropeptide Y (NPY), and their analogues (Bloom, SR, et al. (2008) Mol Interv 8:82-98; Field, BC, et al. (2009) Br J Clin Pharmacol 68:830-843).

[0283] Within the scope of the embodiments presented herein, methods for treating obesity are also considered, which include the administration of peptide products described herein in combination with peptide hormones, including but not limited to leptin, ghrelin, and CART (cocaine-modulated transcript and amphetamine-modulated transcript) analogs and antagonists.

[0284] Additional peptide products throughout the body are known to be associated with adipocytes or obesity (adipokines) and have pro-inflammatory effects (Gonzalez-Periz, A. and Claria, J. (2010) Scientific World Journal 10:832-856). Such agents, when used in combination with the peptide products described herein, will have additional desirable effects. Examples of agents that provide beneficial effects when used in combination with the peptide products described herein include analogues and antagonists of adiponectin, chemerin, visfatin, nesfatin, omentin, resistin, TNF-alpha, IL-6, and obestatin.

[0285] <Intermediate> In one embodiment, the Specified Provision provides intermediates and / or reagents comprising a surfactant moiety and a reactive functional group capable of forming a single bond with a reactive functional group on a natural or unnatural amino acid. Such intermediates and / or reagents enable improvements in the bioavailability of peptides and / or proteins used in human and animal diseases, and in the pharmacokinetic and / or pharmacodynamic behavior of pharmaceuticals. Covalent bonding of such intermediates and / or reagents via functional groups on the side chains of amino acids, for example, on the epsilon-amino function of Lys, on the sulfhydryl of Cys, or at the amino or carboxyl terminus of peptide and / or protein targets enables the synthesis of peptide products described herein. In specific embodiments, the nonionic surfactant moiety is a monosaccharide or disaccharide having an O-alkyl glycoside substitution, wherein the glycosidic bond has an alpha or beta structure. In specific embodiments, the O-alkyl chain is C1-C 20 , or C6-C 16 It is an alkyl chain.

[0286] In another embodiment, the Specified Provisions Provide an intermediate and / or reagent comprising a nonionic surfactant moiety having a specific alkyl glycosidic bond that mimics an O-alkyl glycosidic bond, and a reactive functional group capable of forming a single bond with a reactive functional group on a natural or unnatural amino acid. Such intermediate and / or reagents incorporate an S-linked alkyl chain or an N-linked alkyl chain, and have altered chemical and / or enzymatic stability compared to products to which an O-linked alkyl glycoside is bonded.

[0287] In some embodiments, the intermediates and / or reagents provided herein are compounds in which the hydrophilic group is modified glucose, galactose, maltose, glucuronic acid, diglucuronic acid, etc. In some embodiments, the hydrophilic group is glucose, maltose, glucuronic acid, or diglucuronic acid, and the hydrophobic group is C1-C 20The alkyl or aralkyl chain is present. In some embodiments, the glycosidic bond to the hydrophobic group is alpha-configured, and in some cases, this bond is beta-configured at the anomeric center on the saccharide.

[0288] In some embodiments, the hydrophilic group is glucose, maltose, glucuronic acid, or diglucuronic acid, and the hydrophobic group is C1-C 20 It is an alkyl chain or aralkyl chain.

[0289] In some embodiments, the intermediates and / or reagents provided herein include surfactants that contain reactive functional groups, such as carboxylic acid groups, amino groups, azides, aldehydes, maleimides, sulfhydryls, hydroxylamino groups, and alkynes.

[0290] In some embodiments, the intermediate and / or reagent is an O-linked alkyl glycoside having one hydroxyl group modified to be a carboxylic acid functional group or an amino functional group. In some embodiments, the reagent is a 1-O-alkylglucuronic acid of alpha or beta configuration, where the alkyl chain is C1 to C 20 The length is C6 to C 16 It is the length.

[0291] In some embodiments, the reagent is a 1-O-alkyldiglucuronic acid having an alpha or beta configuration, where the alkyl chain is C1 to C 20 The length is C6 to C 16 It is the length.

[0292] In some embodiments, the reagent is an alpha or beta-constituent S-bonded alkyl glycoside having one hydroxyl group that is modified to become a carboxylic acid functional group or an amino functional group.

[0293] In some embodiments, the reagent is an N-linked alkyl glycoside of alpha or beta configuration having one hydroxyl group that is modified to become a carboxylic acid functional group or an amino functional group.

[0294] In another embodiment, this specification provides peptide and / or protein products containing covalently bonded alkyl glycosides that have properties acceptable for use in human and animal diseases. Schematic Figure 1 lists typical nonionic surfactants that can be modified to yield reagents and / or intermediates useful for the synthesis of peptide products modified with the surfactants described herein.

[0295] [ka]

[0296] In some embodiments, the covalently modified peptides and / or proteins described herein incorporate a surfactant moiety into the peptide structure. In specific embodiments, the covalently modified peptides and / or proteins described herein incorporate nonionic surfactants of the class alkyl, optionally substituted alkyl, alkoxyaryl, or aralkyl glycoside. Alkyl glycosides are important products and are widely used in the food, service, and cleaning industries. Therefore, their production on a commercially significant scale has been the subject of extensive research. Both enzymatic and chemical processes are available for production at very low cost (Park, DW, et al. (2000) Biotechnology Letters 22:951-956). Additional examples of well-known routes to 1-alkyl glycosides using acid-catalyzed glycosylation or Koenigs-Knorr coupling are shown in "Milkereit, G., et al. (2004) Chem Phys Lipids 127:47-63; Vill, V., et al. (2000) Chem. Phys. Lipids 104, 75-9; Helferich, B., et al. (1956) Chem. Ber. 89, 314-315," and the references cited therein. In the case of optionally substituted alcohols, protection of the functional group (preferably an acid-unstable one, such as an ester like t-butyl or p-methoxybenzyl) is introduced by means known to those skilled in the art (see Greene, T. and Wuts, PGM (1999) Protective Groups in Organic Synthesis, John Wiley & Sons, Inc. and the references therein). For example, protection of 16-hydroxyhexadecanoic acid by formation of a t-butyl ester using N,N-dimethylformamide di-tert-butyl acetal or a similar reagent yields t-butyl 16-hydroxyhexadecanoate for glycosylation of a desired acetobrom sugar catalyzed by a mercury or silver salt.

[0297] Such alkyl and optionally substituted alkyl glycosides can be further modified to produce intermediates for the synthesis of covalently modified peptides and / or proteins as described herein. Therefore, it is known that using unprotected materials and a platinum black catalyst in the presence of oxygen, 1-dodecyl beta-D-glucoside is preferentially oxidized at the 6-position, thereby yielding the corresponding glucuronic acid analog in high yield (van Bekkum, H. (1990) Carbohydrates as Organic Raw Materials 289-310). Additional and chemoselective methods are available for the oxidation of primary alcohols at the 6-position of alkyl glucosides. For example, the catalytic use of 2,2,6,6-tetramethyl-1-piperidinyloxyl (TEMPO) with a stoichiometric amount of the organic oxidizing agent [bis(acetoxy)iodo]benzene (BAIB) (De Mico, A., et al. (1997) J Org Chem 1997:6974-6977) yielded a remarkable yield of nucleoside-5'-carboxylic acid (Epp, J.B. and Widlanski, TS (1999) J Org Chem 64:293-295) by oxidation of the primary hydroxyl. This oxidation is performed when other secondary hydroxyls are not protected and is chemoselective to the primary hydroxyl (Codee, J.D., et al. (2005) J Am Chem Soc 127:3767-3773). Similarly, 1-dodecyl β-D-glucopyranoside, 1-tetradecyl·β-D-glucopyranoside, 1-hexadecyl β-D-glucopyranoside, 1-octadecyl β-D-glucopyranoside, and 1-aicosyl β-D-glucopyranoside were oxidized in water by TEMPO using stoichiometric amounts of KBr and sodium hypochlorite (Milkereit, G., et al. (2004) Chem Phys Lipids 127:47-63) as oxidizing agents to the corresponding uronic acids (1-dodecyl β-D-glucuronic acid, 1-hexadecyl β-D-glucuronic acid, 1-octadecyl β-D-glucuronic acid, and 1-aicosyl β-D-glucuronic acid).A mild oxidation procedure using (diacetoxyiodo)benzene (DAIB aka BAIB) is provided in the examples. Certain such glucuronic acid intermediates are commercially available (e.g., octyl bD glucuronic acid; Carbosynth, MO 07928), and as shown, a wide range of them are subject to preparation by conventional methods (Schamann, M. and Schafer, HJ (2003) Eur J Org Chem 351-358; Van den Bos, LJ, et al. (2007) Eur J Org Chem 3963-3976), or, if necessary, from commercial sources. Schematic Figure 2 shows, as an example, certain functionalized surfactant intermediates containing a -COOH group as a reactive functional group used to prepare the intermediates and / or reagents described herein.

[0298] [ka]

[0299] Similarly, aralkyl glycosides (including alkoxyaryls) can form the basis for reagents of closely related nonionic surfactants. For example, 4-alkoxyphenyl β-D-glucopyranoside is readily synthesized by the reaction of 4-alkyloxyphenol with penta-O-acetyl β-D-glucose in the presence of boron trifluoride etherate. Deacetylation after using trimethylamine in methanol / water, and selective oxidation as described above, yield reagents of alkoxyaryl glucuronic acid suitable for forming the reagents and peptides described herein, for example ((Smits, E., et al. (1996) J Chem Soc, Perkin Trans I 2873-2877; Smiths, E., et al. (1997) Liquid Crystals 23:481-488).

[0300] [ka]

[0301] The intermediate glucuronic acid class is readily activated by standard coupling agents for bonding to amino acid side chains, such as the Lys side chain. Therefore, Fmoc-Lys-O-TMS (trimethylsilyl=TMS) can react with octyl beta-D-glucuronic acid in the presence of a coupling agent, and the O-TMS protecting group can be hydrolyzed in an aqueous workup, as shown schematicly in Figure 4, to yield Fmoc-Lys (1-octyl beta-D-glucuronamide). This reagent can be used for incorporation into solid-phase peptide synthesis using standard coupling protocols when it is desired to incorporate the surfactant moiety near the N-terminal region of the molecule. The secondary hydroxyl groups may remain unprotected due to the high reactivity of the Lys amino functional groups, or because they can be protected by peracetylation. When the acetyl-protected form is used, the acetyl-protecting group can be removed in high yield by treatment with MeOH / NaOMe or MeOH / Et3N. Schematic Figure 4 illustrates the preparation of the reagents described herein.

[0302] [ka]

[0303] In some embodiments, the reagents and / or intermediates for the preparation of the bioactive peptide products described herein include a family of linker amino acids modified with surfactants for incorporation into the synthetic peptide product. Thus, in one embodiment, the peptide product described herein is synthesized linearly, where a functionalized surfactant is bonded to a linker amino acid reversibly protected by a functional group on the side chain of the linker amino acid (e.g., an amino group on a lysine residue) to obtain a proprietary reagent (as shown in schematic Figure 4), which can be incorporated into the growth peptide chain, and the remaining peptide is then synthesized by further amino acid bonding to a cysteine ​​residue. Protecting groups suitable for the synthesis of the modified peptides and / or proteins described herein are, for example, described in TW Green, PGM Wuts, Protective Groups in Organic Synthesis, Wiley-Interscience, New York, 1999, 503-507, 736-739, the disclosure of which is incorporated herein by reference.

[0304] In another embodiment, the peptide products described herein are synthesized by covalent bonding of a functionalized surfactant to a full-length peptide via an appropriate functional group on a linker amino acid in the peptide chain.

[0305] Alternatively, functionalized surfactants can be added to the deprotected linker amino acid side chain during solid-phase synthesis of peptides. For example, alkylglucuronyl groups can be directly added to the linker amino acid side chain (e.g., the deprotected Lys side chain) during solid-phase synthesis of peptides. For instance, the use of Fmoc-Lys(Alloc)-OH as a subunit provides orthogonal protection that can be removed while the peptide is still on the resin. Thus, deprotection of the Lys side chain using Pd / thiobarbital, Pd / 1,3-dimethylbarbituric acid (DMBA), or other Alloc deprotection recipes allows for the exposure of the amino group for coupling with acyl-protected or unprotected 1-octylbeta-D-glucuronic acid units or for side-chain lactam formation. Final deprotection with a low % CF3CO2H(TFA) cleavage cocktail then delivers the desired product. Although glycosidic bonds are unstable in strong acids, it has been obtained herein or elsewhere that they are relatively stable under low % TFA cleavage conditions. Alternatively, acyl protection (e.g., acetyl, Ac; benzoyl, Bz) or trialkylsilyl protection on the saccharide OH functional group can be used to provide increased protection for the glycosidic bond. Subsequent deprotection with a base (NH2NH2 / MeOH; NH3 / MeOH, NaOMe / MeOH) yields the desired deprotected product. Schematic Figure 4 illustrates the reagents described herein. Schematic Figure 5 illustrates an unrestricted example of a peptide intermediate described herein. This example illustrates a peptide having a surfactant bond at the N-terminus of the peptide, but the methods described herein are suitable for the synthesis of peptide intermediates having a surfactant bond in the intermediate region, C-terminal region, or at any position within the peptide.

[0306] [ka]

[0307] As shown below in schematic Figure 6, additional reagents are generated by modification of the 6-position functional group, providing various means of bonding the amino acid side chain to the functional group. Thus, amino substitutions can be used for bonding to Asp or Glu side chains. Azide or alkyne substitutions can be used for bonding to non-natural amino acids containing complementary acceptors for Huisgen 3+2 cycloaddition (Gauthier, MA and Klok, HA (2008) Chem Commun (Camb) 2591-2611). Aminooxy or aldehyde functional groups can be used for bonding to aldehydes (i.e., oxime bonding) or to amino functional groups (i.e., reductive alkylation), respectively. Maleimide or -NH-(C=O)-CH2-Br functional groups can be chemoselectively bonded to Cys or other SH functional groups. These types of bonding strategies are advantageous when used in conjunction with the reagents described herein. Interconversion of functional groups is widely practiced in organic synthesis, and a comprehensive list of multiple pathways to each of the functional group modifications listed herein is available ((Larock, RC (1999)) "Comprehensive Organic Transformations", VCH Publishers, New York).

[0308] Therefore, for example, the primary hydroxyl at position 6 of octyl 1-β-D-glucoside is converted to an azide by reactions such as activation and substitution with azide anions, or reactions used in carbohydrate chemistry (e.g., by tosylation and subsequent NaN3). The corresponding azide is reduced to an amino functional group by reduction with thioacetic acid in pyridine (Elofsson, M., et al. (1997) Tetrahedron 53: 369-390) or by a similar method of amino group formation (Stangier, P., et al. (1994) Liquid Crystals 17: 589-595). Approaches to the acetylene, aminooxy, and aldehyde moieties are best carried out in the triacetoxy form, which is available from commercially available glucosides, by treatment with Ac2O and subsequent mild hydrolysis of the primary amine. This 6-hydroxy form can be selectively oxidized to an aldehyde or activated as a tosylate or triflate and replaced by NH2OH or sodium acetylide. The maleimide bond may be via a carbon bond as shown, or preferably via an O or amide bond, again by the coupling of a glucuronic acid derivative to an activated hydroxyl substitution or amino-bonded maleimide reagent, as is well known in the art. Further functional group transformations are well known to those skilled in medicinal chemistry and are within the scope of the embodiments described herein.

[0309] Furthermore, within the scope of the synthetic methods described herein, surfactants are considered, where the saccharide and hydrophobic chains are covalently bonded via α-glycosidic bonds. Synthetic routes to predominantly α-bonded glycosides are well known in the art and typically begin with peracetyl sugar, achieving α-glycosylation using acidic catalysis (e.g., SnCl4, BF3, or HCl) (Cudic, M. and Burstein, GD (2008) Methods Mol Biol 494: 187-208; Vill, V., et al. (2000) Chem Phys Lipids 104: 75-91 (incorporated herein by reference for such disclosure)). A similar synthetic route exists for disaccharide glycosides (von Minden, HM, et al. (2000) Chem Phys Lipids 106: 157-179 (incorporated herein by reference for such disclosure)). Functional group transformations are then carried out to lead to 6-carboxylic acids, etc., for the production of the corresponding α-bonded reagents, as described above.

[0310] Schematic Figure 6 lists specific compounds and reagents useful for the synthesis of covalently modified peptides and / or proteins described herein. Standard notation using single-letter abbreviations for amino acids is used.

[0311] [ka]

[0312] Many alkyl glycosides can be synthesized by known procedures, for example, as described in (Rosevear, P., et al. (1980) Biochemistry 19: 4108-4115, Li, YT, et al. (1991) J Biol Chem 266: 10723-10726) or Koeltzow and Urfer, J. Am. Oil Chem. Soc., 61:1651-1655 (1984), U.S. Patent Application No. 3,219,656 and U.S. Patent Application No. 3,839,318, or enzymatically, for example, as described in (Li, YT, et al. (1991) J Biol Chem 266: 10723-10726, Gopalan, V., et al. (1992) J Biol Chem 267:9629-9638). O-alkyl bonding to natural amino acids such as Ser can be carried out on Fmoc-Ser-OH using peracetylglucose to produce Nα-Fmoc-4-O-(2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl)-L-serine. This substance can be selectively deprotected at the primary carbon atom (position 6) as described above and selectively oxidized using TEMPO / BAIB to produce the corresponding 6-carboxyl functional group, which can be bonded to lipophilic amines to generate a new class of nonionic surfactants and reagents (Schematic Figure 7).

[0313] [ka]

[0314] The bond between hydrophobic alkyls and hydrophilic saccharides can include, among others, glycosides, thioglycosides, amides (Carbohydrates as Organic Raw Materials, FW Lichtenthaler ed., VCH Publishers, New York, 1991), ureids (Austrian Pat. 386,414 (1988); Chem. Abstr. 110:137536p (1989); see Gruber, H. and Greber, G., "Reactive Sucrose Derivatives" in Carbohydrates as Organic Raw Materials, pp. 95-116), or ester bonds (Sugar Esters: Preparation and Application, JC Colbert ed., (Noyes Data Corp., New Jersey), (1974)).

[0315] Examples of alkyl glycosides that may be selected for use in mutations to the reagents described herein or for the formulation of the products include: alkyl glycosides such as octyl-, nonyl-, decyl-, undecyl-, dodecyl-, tridecyl-, tetradecyl-, pentadecyl-, hexadecyl-, heptadecyl-, and octadecyl-D-maltoside, -melibioside, -glucoside or -sucroside (i.e., sucrose esters) (synthesized according to Koeltzow and Urfer; Anatrace Inc., Maumee, Ohio; Calbiochem, San Diego, Calif.; Fluka Chemie, Switzerland); alkylthiomaltosides such as heptyl-, octyl-, dodecyl-, tridecyl-, and tetradecyl-β-D-thiomaltoside; (Carbohydrates as Organic Raw Materials, 247-265 (FW Lichtenthaler, ed.) VCH Publishers, New York (1991); synthesized according to Defaye, J. and Pederson, C., “Hydrogen Fluoride, Solvent and Reagent for Carbohydrate Conversion Technology” in Ferenci, T., J. Bacteriol, 144:7-11 (1980); alkylthioglucosides such as 1-dodecyl- or 1-octyl-thioα- or β-D-glucopyranoside (see Anatrace, Inc., Maumee, Ohio; Saito, S. and Tsuchiya, T. Chem. Pharm. Bull. 33:503-508 (1985)); alkylthiosucrose (e.g., synthesized according to Binder, TP and Robyt, JF, Carbohydr. Res. 140:9-20 (1985)); alkylmaltotriosides (Koeltzow and Synthesized according to Urfer; Long-chain aliphatic carbonate amide of sucrose amino-alkyl ether (Austrian Patent 382,381 (1987); Chem. Abstr., synthesized according to 108:114719 (1988) and Gruber and Greber pp. 95-116); derivatives of palatinose and isomaltamine linked by amide bonds to alkyl chains (synthesized according to Kunz, M., "Sucrose-based Hydrophilic Building Blocks as Intermediates for the Synthesis of Surfactants and Polymers" in Carbohydrates as Organic Raw Materials, 127-153); derivatives of isomaltamine linked to alkyl chains by urea (synthesized according to Kunz); long-chain aliphatic carbonate ureids of sucrose amino-alkyl ethers (synthesized according to Gruber and Greber, pp. 95-116); and long-chain aliphatic carbonate amides of sucrose amino-alkyl ethers (Austrian Patent 382,381 (1987), Chem. Abstr., 108:114719 (1988) and Gruber and (Synthesized according to Greber, pp. 95-116).

[0316] Several preferred glycosides that can be further modified to incorporate reactive functional groups for binding to peptides include maltose, sucrose, glucose, and galactose saccharides linked by glycoside or ester bonding to an alkyl chain of 6, 8, 10, 12, 14, or 16 carbon atoms, e.g., hexyl-, octyl-, decyl-, dodecyl-, tetradecyl-, and hexadecyl-maltosides, -melibiosides, -sucrosides, -glucosides, and -galactosides. In the body, these glycosides are broken down into harmless alcohols or fatty acids and oligosaccharides or saccharides. The above examples are examples of types of alkyl glycosides used in the manner claimed herein, but the list is not intended to be exhaustive.

[0317] Generally, these surfactants (e.g., alkyl glycosides) are optionally designed or selected to modify the biological properties of peptides, such as their bioavailability, half-life, receptor selectivity, toxicity, biodistribution, solubility, and stability, as well as their resistance to enzymatic degradation, structural properties, spectroscopic properties, chemical and / or photochemical properties, catalytic activity, redox potential, and ability to react with other molecules covalently or noncovalently, such as their ability to react with other molecules, for example, covalently or noncovalently.

[0318] <Surfactants> The term "surfactant" is an abbreviation of the phrase "surface active agent." In pharmaceutical applications, surfactants are useful in liquid pharmaceutical formulations and serve many purposes, acting as emulsifiers, solubilizers, and wetting agents. Emulsifiers stabilize aqueous solutions of lipophilic or partially lipophilic substances. Solubilizers increase the solubility of components of pharmaceutical compositions, increasing the concentration that can be achieved. Wetting agents are chemical additives that reduce the surface tension of a fluid, inducing it to spread easily on the surface to which it is applied, and thus even causing "wetting" of the surface by the fluid. Wetting agents provide a means for a liquid formulation to achieve close contact with mucous membranes or other surface areas to which the pharmaceutical formulation comes into contact. Thus, surfactants can be useful additives for modifying the properties of the peptides themselves, in addition to stabilizing the formulations of peptide products described herein.

[0319] In specific embodiments, synthetically available alkyl glycosides, such as the alkyl glycosides dodecyl, tridecyl, and tetradecyl maltoside, -melibioside, or -glucoside, as well as sucrose dodecanoate, tridecanoate, and tetradecanoate, are suitable for covalent bonding to peptides as described herein. Similarly, the corresponding alkylthioglycosides are stable, synthetically available surfactants that are acceptable for formulation development.

[0320] A wide range of physical properties and surfactant properties can be achieved by appropriate modification of the hydrophobic or hydrophilic regions of surfactants (e.g., alkyl glycosides). For example, studies comparing the bilayer activity of dodecyl maltoside (DM) with that of dodecyl glucoside (DG) have found that, despite having hydrophobic tails of the same length, the bilayer activity of DM is more than three times higher than that of DG (Lopez, O., et al. (2002) Colloid Polym Sci 280: 352-357). In this specific example, polar identity (disaccharide versus monosaccharide) affects the behavior of the surfactant. In the case of surfactants conjugated to peptides (e.g., the peptide products described herein), the peptide region can also contribute to the overall hydrophobic or hydrophilic characteristics of the molecule. Accordingly, tuning of physical properties and surfactant properties can be used to achieve specific physical and pharmaceutical properties suitable for an individual peptide target.

[0321] <PEG modification> In some embodiments, the surfactant-modified peptide products described herein are further modified to incorporate one or more PEG moieties (Veronese, F.M. and Mero, A. (2008) BioDrugs 22: 315-329). In some examples, incorporation of large PEG chains prevents filtration of the peptide through glomeruli in the kidney into diluted urine formed therein (Nestor, J.J., Jr. (2009) Current Medicinal Chemistry 16: 4399 - 4418, Caliceti, P. and Veronese, F.M. (2003) Adv Drug Deliv Rev 55: 1261-1277). In some embodiments, the optional hydrophilic PEG chain allows balancing of the solubility and physical properties of peptides or proteins that have been hydrophobized by incorporation of longer-chain alkyl glycoside moieties.

[0322] Protein pegylation can also have potentially negative effects. Therefore, pegylation can lead to a significant loss of biological activity for some proteins, which may be related to ligands for specific classes. In such cases, reversible pegylation may be beneficial (Peleg-Shulman, T., et al. (2004) J Med Chem 47: 4897-4904, Greenwald, RB, et al. (2003) Adv Drug Deliv Rev 55: 217-250, Roberts, MJ and Harris, JM (1998) J Pharm Sci 87: 1440-1445).

[0323] Furthermore, increased molecular weight may prevent permeation through physiological barriers other than the glomerular membrane barrier. For example, it has been suggested that high molecular weight PEGylated forms may prevent permeation into certain tissues, thereby reducing therapeutic efficacy. In addition, high molecular weight may prevent uptake across mucosal membrane barriers (nasal, buccal, vaginal, oral, rectal, and pulmonary delivery). However, delayed uptake can be highly advantageous for the delivery of stable molecules to the lungs, substantially extending the duration of action. The peptide and / or protein products described herein increase bioavailability in the oral mucosa, which allows longer-chain PEG modifications to be used in combination with surfactant modifications, along with achieving commercially significant bioavailability via intranasal or other oral mucosal pathways.

[0324] In some embodiments, long-chain and short-chain PEG polymers are suitable for the modification of proteins and peptides described herein. Inhalation-based treatment for diabetes is a novel approach for drug delivery, and the lungs have a highly permeable barrier (e.g., Exubera). For this application, delayed permeation of the lung barrier is preferred as a form of PEGylation. 10 ~C 400The molecular weight ranges from approximately 250 Da to 10,000 Da. Therefore, the primary pathway to elongation by PEG is achieving an "effective molecular weight" above the glomerular filtration cut-off (above 68 kDa), although the use of shorter chains may be a pathway for prolonging retention in the lungs for the treatment of lung diseases and other respiratory diseases. Thus, PEG chains of about 500 Da to 3000 Da are sufficient to delay entry into the peripheral circulation, but are insufficient in size to obtain significantly prolonged circulation times. In some embodiments, pegylation is applied to increase local efficacy in lung tissue with a reduced likelihood of systemic side effects related to the covalently modified peptides and / or proteins described herein. In some such embodiments, PEG chains in the range of about 750 Da to about 1500 Da are collectively referred to as "PEG1K".

[0325] Furthermore, other polymers may be used in combination with the compounds described herein to optimize their physical properties. For example, poly(2-ethyl2-oxazoline) conjugates have variable hydrophobicity and sufficient size to extend the duration of action (Mero(A) et al. (2008)J control release 125:87-95). The binding of such polymers to saccharides produces a class of surfactants suitable for use in the modification of peptides and / or proteins described herein.

[0326] Polyethylene glycol chains are functionalized to enable their conjugation to reactive groups on peptide and / or protein chains. Typical functional groups enable the reaction of amino, carboxyl, or sulfhydryl groups on peptides with corresponding carboxyl, amino, or maleimide groups (etc.) on the polyethylene glycol chain. In one embodiment, PEG is C 10 -C 3000The chain is included. In another embodiment, PEG has a molecular weight greater than 40,000 daltons. In yet another embodiment, PEG has a molecular weight less than 10,000 daltons. PEG as a protein modification is well known in the art, and its uses are described, for example, in U.S. Patent Applications No. 4,640,835; No. 4,496,689; No. 4,301,144; No. 4,670,417; No. 4,791,192; and No. 4,179,337.

[0327] Non-traditional types of PEG chains are modified to be naturally amphiphilic; that is, they are modified to contain hydrophobic regions such as fatty acid esters and other hydrophobic components, while still possessing a hydrophilic PEG structure. See, for example (Miller, MA, et al. (2006) Bioconjug Chem 17: 267-274); Ekwuribe, et al. US 6,309,633; Ekwuribe, et al. US 6,815,530; Ekwuribe, et al. US 6,835,802). These amphiphilic PEG conjugates for proteins were originally developed to increase oral bioavailability, but were relatively ineffective in this role. However, the use of such amphiphilic PEG conjugates with amphiphilic peptides significantly extends retention in the lungs and extends the useful bioactivity of these drugs. Preferred PEG chains are in the molecular weight range of 500 Da to 3000 Da. The methods for synthesizing these conjugates are described in detail in the above-mentioned references, and their entire contents are incorporated herein by reference.

[0328] PEG itself does not possess functional groups that can bind to target molecules such as peptides. Therefore, in order to create a PEG bond, the PEG entity must first be functionalized, and then the functionalized bond is used to attach the PEG entity to the target molecule, such as a peptide (Greenwald, RB, et al. (2003) Adv Drug Deliv Rev 55: 217-250, Veronese, FM and Pasut, G. (2005) Drug Discov Today 10: 1451-1458, Roberts, MJ, et al. (2002) Adv Drug Deliv Rev 54: 459-476). In one embodiment, site-specific pegylation can be achieved by cysteine ​​substitution on the peptide molecule. As described herein, the target peptide can be synthesized by solid-phase synthesis, recombinant means, or other means.

[0329] Accordingly, in some embodiments, the peptide products described herein include a Lys or alkyl glycoside and at least one Cys residue, a Lys residue, or other reaction residues modified by specific pegylation on other reaction amino acid residues anywhere in the molecule.

[0330] In another embodiment, Lys or other residues having nucleophilic side chains may be used to incorporate PEG residues. This can be achieved by using the attachment of amides or carbamates to the PEG-carboxyl or PEG-carbonate chains, see, for example, (Veronese, FM and Pasut, G. (2005) Drug Discov Today 10: 1451-1458). An alternative approach is to modify the amino functional group of the Lys side chain by attaching SH-containing residues such as mercaptoacetyl, mercaptopropionyl (CO-CH2-CH2-CH2-SH). Alternatively, the PEG chain may be incorporated at the C-terminus as an amide during the course of synthesis. Additional methods for attaching the PEG chain utilize the reaction with His and Trp side chains. Other similar methods for modifying peptide chains to enable PEG chain binding are known in the art and are incorporated herein by reference (Roberts, MJ, et al. (2002) Adv Drug Deliv Rev 54: 459-476).

[0331] <Pharmaceutical Products> In one embodiment, a covalently modified peptide or protein as described herein is provided in a formulation that, when administered to a subject, further reduces, prevents, or mitigates the association or aggregation of peptides and / or proteins in the composition, for example, by reducing the self-association or self-aggregation of peptides and / or proteins, or by reducing the association or aggregation with other peptides or proteins.

[0332] Self-aggregation at high protein concentrations is a problem in therapeutic formulations. For example, self-aggregation increases the viscosity of concentrated monoclonal antibodies in aqueous solutions. Concentrated insulin preparations are inactivated by self-aggregation. The interactions of these self-aggregating proteins reduce, modulate, or eliminate the biological activity of many therapeutic agents, especially at high protein concentrations (Clodfelter, DK, et al. (1998) Pharm Res 15: 254-262). Therapeutic proteins formulated at high concentrations for delivery by injection or other means may be physically unstable or become insoluble as a result of interactions between these proteins.

[0333] A major challenge in the preparation of peptide and protein formulations is developing manufacturable and stable dosage forms. Physical stability properties, crucial for processing and handling, are often poorly characterized and difficult to predict. Various physical instability phenomena occur, such as aggregation, aggregation, crystallization, and precipitation, as determined by protein interaction and solubility properties. This results in significant challenges in manufacturing, stability, analysis, and delivery. The development of formulations for peptide and protein drugs requiring high doses (in mg / kg units) is needed in many clinical settings. For example, using the SC route, approximately <1.5 mL is an acceptable dose. To achieve adequate dosing, this may require a protein concentration of >100 mg / mL. Similar considerations exist for the development of high-concentration lyophilized formulations for monoclonal antibodies. Generally, higher protein concentrations result in smaller injection volumes, which is crucial for patient comfort, convenience, and compliance. The surfactant-modified compounds described herein are designed to minimize such aggregation events, which can be further facilitated through the use of small amounts of surfactant as described herein.

[0334] Because injection is an unpleasant mode of administration for many people, other means of administering peptide therapeutics are needed. For example, certain peptide and protein therapeutics can be administered intranasally, buccally, orally, vaginally, by inhalation, or by other oral mucosal administration. Examples include nafarelin (Synarel®) and calcitonin, which are administered as commercially available nasal sprays. The covalently modified peptides and / or proteins described herein are designed to facilitate such oral mucosal administration, and such formulations may be further facilitated through the use of small amounts of surfactants as described herein.

[0335] Typical formulation parameters include the selection of optimal solution pH, buffer, and stable excipients. Furthermore, the reconstitution of lyophilized cakes is crucial for lyophilized or powdered formulations. A further and significant problem involves changes in the viscosity of protein formulations during self-assembly. Changes in viscosity can significantly alter delivery characteristics, such as spray (aerosol) delivery for intranasal, pulmonary, or oral sprays. Additionally, increased viscosity can make injection delivery via syringe or intravenous line more difficult or impossible.

[0336] Numerous attempts have been reported to stabilize and maintain the integrity and physiological activity of peptides. In particular, some attempts have provided stabilization against thermal denaturation and aggregation for insulin pump systems. High molecular weight surfactants have been described (Thurow, H. and Geisen, K. (1984) Diabetologia 27: 212-218; Chawla, AS, et al. (1985) Diabetes 34: 420-424). The stabilization of insulin by these compounds was thought to be due to the stabilization of its steric properties. Other systems used include saccharides (Arakawa, T. and Timasheff, SN (1982) Biochemistry 21: 6536-6544), osmoregulators such as amino acids (Arakawa, T. and Timasheff, SN (1985) Biophys J 47: 411-414), and water structure breakers such as urea (Sato, S., et al. (1983) J Pharm Sci 72: 228-232). These compounds exert their effects by regulating hydrophobic interactions within proteins or peptide molecules.

[0337] Various peptides, proteins, or other peptides can be modified with any of the covalently bonded surfactant reagents described herein. Conveniently, the peptide modifications described herein involve covalent bonding of surfactants containing both hydrophilic (e.g., saccharide) and hydrophobic (e.g., alkyl chain) groups, thereby enabling the stabilization of peptides under physiological conditions. In some embodiments, the covalent bonding of hydrophilic and hydrophobic (e.g., glycoside surfactant) moieties to peptides and / or proteins described herein eliminates the need to modify the amino acid sequence of the peptide and / or protein, thereby enhancing stability (e.g., reducing aggregation).

[0338] In some embodiments, the formulation comprises at least one drug comprising a peptide modified with a surfactant-derived reagent described herein, which may be further bound in the formulation with a surfactant, where the surfactant is further composed of, for example, a saccharide, an alkyl glycoside, or other excipients, and may be administered in a format selected from the group consisting of infusion, spray, aerosol, lyophilized product, spray-dried product, injection, and sustained-release format. Sprays and aerosols may be achieved through the use of a suitable dispenser and may be administered by intranasal, transbuccal, inhalation, or other oral mucosal routes. Lyophilized products may contain other compounds such as mannitol, saccharide, submicron anhydrous α-lactose, gelatin, biocompatible gel, or polymer. Sustained-release formats may include eyeball inserts, erosive microparticles, hydrolyzable polymers, swollen mucosal-adhering particles, pH-sensitive microparticles, nanoparticle / latex systems, ion-exchange resins, and other polymeric gels and implants (Ocusert, Alza Corp., California; Joshi, A., S. Ping and KJ Himmelstein, Patent Application WO 91 / 19481). Significant oral bioavailability is also achievable.

[0339] The peptide and protein modifications described herein may mitigate, and in some cases eliminate, the need for organic solvents. Trehalose, lactose, mannitol, and other saccharides have been used to prevent aggregation. Aggregation of humanized anti-IgE monoclonal antibodies has been minimized by formulations with trehalose in molar ratios ranging from 300:1 to 500:1 (excipient:protein) or greater. However, powders were either excessively agglutinative and unsuitable for aerosol administration or exhibited undesirable protein glycation during storage (Andya, JD, et al. (1999) Pharm Res 16: 350-358). Each of the discovered additives has limitations, such as xenobiotic metabolism, irritation or toxicity, or high cost. For use with the covalently modified peptides and / or proteins described herein, excipients are considered that are effective, non-irritating, and non-toxic, composed of natural sugars, fatty acids, or long-chain alcohols and therefore do not require xenobiotic metabolism, and can be used to minimize aggregation in aqueous solutions or in aqueous reconstitution of dried peptide and / or protein formulations in situ by physiological aqueous reconstitution with aqueous bodily fluids such as plasma or saliva.

[0340] Other formulation components may include, among other things, buffers and physiological salts, non-toxic protease inhibitors such as aprotinin and soy trypsin inhibitors, α1-antitrypsin, and monoclonal antibodies that inactivate proteases. Buffers may include organic substances such as acetates, citrates, glucons, fumarates, malates, polylysine, polyglutamates, chitosan, and dextran sulfates, or inorganic substances such as phosphates and sulfates. Such formulations may further contain small concentrations of bacteriostatic agents such as benzyl alcohol.

[0341] Formulations suitable for intranasal administration also include solutions or suspensions of modified peptide and / or protein products described herein in an acceptable evaporative solvent such as a hydrofluoroalkane. Such formulations are suitable for administration from a metered inhaler (MDI) and have the advantages of no migration from the administration site, low irritation, and no need for sterilization. Such formulations may also contain acceptable excipients or expanders such as submicron anhydrous α-lactose.

[0342] In yet another embodiment, the covalently modified peptides and / or proteins described herein exhibit an increased shelf life. As used herein, the phrase “shelf life” is broadly defined as the length of time during which the product can be stored without becoming unsuitable for use or consumption. The “shelf life” of a composition described herein may also indicate a length of time corresponding to the allowable loss of quality of the composition. A compositional shelf life as used herein is distinguished from an expiration date; “shelf life” relates to the quality of the composition described herein, while “expiration date” relates more to the requirements of manufacturing and testing the composition. For example, a composition past its “expiration date” may still be safe and effective, but its optimal quality is no longer guaranteed by the manufacturer.

[0343] <Medication> The covalently modified peptides and / or proteins described herein may be administered in amounts that provide beneficial therapeutic effects in many disease conditions. In some embodiments, the covalently modified peptides and / or proteins described herein are useful in treating inflammation. In one embodiment, the compounds described herein provide beneficial activity in regulating postoperative or chronic pain. In one embodiment, the peptide is administered to the patient at concentrations higher or lower than those of other forms used to regulate pain. In yet another embodiment, the peptide is administered with other compounds to produce a synergistic therapeutic effect.

[0344] Typical delivery regimens include oral, oral mucosal administration, parenteral (including subcutaneous, intraperitoneal, intramuscular, and intravenous injection), rectal, buccal (including sublingual), transdermal, inhalation, ophthalmic, and oral mucosal (including intranasal) administration. An attractive and widely used method for peptide delivery involves subcutaneous injection of controlled-release injectable formulations. In some embodiments, the covalently modified peptides and / or proteins described herein are useful for subcutaneous, intranasal, and inhalation administration. Furthermore, depending on the disease being treated, these therapeutic compositions are administered systemically or topically. Formulations and techniques for administration can be found in the latest edition of “Remington's Pharmaceutical Sciences” (Mack Publishing Co, Easton Pa.).

[0345] The selection of the precise dosage and composition, and the most appropriate delivery regimen, is influenced, among other things, by the pharmacological properties of the selected peptide, the nature and severity of the disease being treated, and the recipient's health status and mental clarity. Furthermore, the route of administration consequently leads to differences in the amount of substance absorbed. Bioavailability for peptide administration via different routes is particularly variable, with amounts observed ranging from less than 1% to nearly 100%. Typically, bioavailability from routes other than intravenous, intraperitoneal, or subcutaneous injection is less than 50%.

[0346] Generally, the covalently modified peptides and / or proteins, or salts thereof, described herein are administered by subcutaneous injection at a dose of approximately 0.1 μg / kg to 1000 μg / kg body weight per day or approximately 0.1 μg / kg to 100 μg / kg body weight per day. For a 50 kg human female subject, the daily dose of the active ingredient is approximately 5 μg to 5000 μg or approximately 5 μg to 5000 μg by subcutaneous injection. Different doses may be required depending on the route of administration, the efficacy of the compound, the pharmacokinetic profile, and the applicable bioavailability. By inhalation, the daily dose is 1000 μg to approximately 20,000 μg twice daily. Higher doses may be required in other mammals, such as horses, dogs, and cattle. This dose may be delivered to conventional pharmaceutical compositions by single dose, by multiple applications, or via controlled release, as necessary to achieve the most effective results.

[0347] Pharmaceutically acceptable salts preserve the desired biological activity of the parent peptide without toxic side effects. Examples of such salts include (a) acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.; and salts formed with organic acids, such as acetic acid, trifluoroacetic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, etc.; (b) base addition salts or complexes formed with polyvalent metal cations, such as zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, nickel, cadmium, etc.; or organic cations formed from N,N'-dibenzylethylenediamine or ethylenediamine; or (c) combinations of (a) and (b), such as zinc tannate salts.

[0348] In addition, in some embodiments, pharmaceutical compositions comprising a covalently modified peptide and / or protein as described herein, or a pharmaceutically acceptable salt thereof, as an active ingredient, combined with a pharmaceutically acceptable, non-toxic carrier, are considered. As mentioned above, such compositions may be prepared for parenteral (subcutaneous, intramuscular, or intravenous) administration, particularly in the form of a liquid solution or suspension; for oral or buccal administration, particularly in the form of a tablet or capsule; for intranasal administration, particularly in the form of a powder, nasal drop, evaporative solution, or aerosol; for inhalation, particularly in the form of a liquid solution or dry powder with excipients, as broadly defined; and for rectal or transdermal administration.

[0349] The compositions may be conveniently administered in unit dosage forms and may be prepared by any method well known in the art of pharmaceuticals, for example, as described in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., (1985) (incorporated herein by reference). Formulations for parenteral administration may contain, as excipients, sterile water or saline, alkylene glycols such as propylene glycol, polyalkylene glycols such as polyethylene glycol, saccharides, vegetable oils, hydrogenated naphthalene, serum albumin nanoparticles (such as those used in Abraxane®, American Pharmaceutical Partners, Inc. Schaumburg IL), etc. With regard to oral administration, formulations may be enhanced by the addition of bile salts or acylcarnitine. Formulations for intranasal administration may be solid or solution in an evaporative solvent such as hydrofluorocarbon, or may contain excipients for stabilization, such as saccharides, surfactants, submicron anhydrous α-lactose or dextran, or may be aqueous or oily solutions for use in the form of nasal drops or measuring sprays. For buccal administration, typical excipients include sugars, calcium stearate, magnesium stearate, and pregelatinized starch.

[0350] When formulated for intranasal administration, absorption across the nasal mucosa can be further enhanced by surfactants, such as glycocholic acid, cholic acid, taurocholic acid, ethocholic acid, deoxycholic acid, chenodeoxycholic acid, dehydrocholic acid, glycodeoxycholic acid, cyclodextrin, etc., in amounts ranging from about 0.1 to 15 weight percent, from about 0.5 to 4 weight percent, or in amounts of about 2 weight percent. An additional class of absorption enhancers reported to show greater efficacy with reduced irritation is the class of alkylmaltosides, such as tetradecylmaltoside (Arnold, JJ, et al. (2004) J Pharm Sci 93: 2205-2213, Ahsan, F., et al. (2001) Pharm Res 18: 1742-1746 and the references therein), all of which are incorporated herein by reference.

[0351] Many formulations exhibit advantages when formulated for inhalation delivery. Adsorption of active peptides to easily dispersed solids such as diketopiperazine, e.g., Technospher particles (Pfutzner, A. and Forst, T. (2005) Expert Opin Drug Deliv 2: 1097-1106), or similar structures, gives formulations that result in rapid initial uptake of the therapeutic agent. Lyophilized powders containing active peptides and excipients, particularly glass particles, are useful for delivery to the lungs due to their excellent bioavailability, for example; see, for example, Exubera® (inhaled insulin by Pfizer and Aventis Pharmaceuticals Inc.). Additional systems for peptide delivery by inhalation have been described (Mandal, TK, Am. J. Health Syst. Pharm. 62: 1359-64 (2005)).

[0352] Delivery of covalently modified peptides and / or proteins described herein to a subject over a long period, for example, from one week to one year, can be achieved by a single dose of a controlled-release system containing a sufficient amount of active ingredient for the desired release period. Various controlled-release systems, such as monolithic or reservoir-type microcapsules, depot implants, polymeric hydrogels, osmotic pumps, vesicles, micelles, liposomes, transdermal patches, iontophoresis devices, and alternative injectable dosage forms, can be used for this purpose. Controlled-release excipients have also been developed for twice-weekly or weekly administration, and for example, protected graft copolymer systems (Castillo, GM, et al. (2012) Pharm Res 29: 306-18) can be used for hydrophobic peptides or hydrophobically modified peptides, such as the peptides of the present invention. Localization of the active ingredient to the desired site of delivery is an additional feature of some controlled-release devices, which may prove beneficial for treating specific disorders.

[0353] One form of controlled-release formulation contains peptides or salts thereof dispersed or encapsulated in a slowly degrading, non-toxic, non-antigenic polymer, such as copro(lactic / glycolic acid) acid, as described in the pioneering study of Kent, Lewis, Sanders, and Tice, U.S. Patent Application No. 4,675,189 (incorporated herein by reference). Compounds, or salts thereof, may also be formulated in cholesterol or other lipid matrix pellets or silastomer matrix implants. Additional sustained-release, depot implant or injectable formulations will become apparent to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson ed., Marcel Dekker, Inc., New York, 1978, and RW Baker, Controlled Release of Biologically Active Agents, John Wiley & Sons, New York, 1987.

[0354] Further forms of controlled-release formulations include solutions of biodegradable polymers, such as copoli(lactic acid / glycolic acid) or block copolymers of lactic acid and PEG, which are bioacceptable solvents injected subcutaneously or intramuscularly to achieve depot formulations. Mixtures of peptides described herein with such polymer formulations are suitable for achieving very long durations of action of the formulations.

[0355] As used herein, “therapeutably effective amount” is interchangeable with “effective amount” for the purposes herein, and is determined by considerations known in the art. The amount must be effective in achieving the desired drug-mediated effect in the treated subject suffering from the disease. The therapeutically effective amount also includes, but is not limited to, appropriate measures selected by those skilled in the art, such as improved survival rate, more rapid recovery, i.e., remission, improvement, or elimination of symptoms, or other acceptable biomarkers or surrogate markers.

[0356] However, it is understood that the specific dosage level and frequency of administration for any particular subject requiring treatment may vary and depend on various factors, including the activity of the specific compound used, its metabolic stability and duration of action, age, weight, health status, sex, diet, mode and timing of administration, rate of secretion, drug combinations, severity of the specific disease, and the host being treated.

[0357] The method of administration is not limited to all embodiments of the compositions described herein, including compositions that reduce or eliminate the immunogenicity of peptides and / or proteins, are non-irritating, have antibacterial or antifungal activity, have increased stability or bioavailability of the drug, reduce bioavailability variability of the drug, avoid first-pass liver clearance, and reduce or eliminate any adverse effects. As used herein, the term “immunogenicity” is the ability of a particular substance or composition or drug to elicit an immune response. The immunogenicity of covalently modified peptides and / or proteins described herein is confirmed by methods known in the art.

[0358] All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent that each independent publication, patent, or patent application is specifically and individually incorporated by reference.

[0359] The covalently modified peptides and / or proteins and reagents for their synthesis described herein are specifically described by the following examples, intended for illustrative purposes only, so that numerous modifications and changes will be apparent to those skilled in the art. [Examples]

[0360] Example 1: Reagent - N-α-Fmoc,N-ε-(1-octylβ-D-glucuronide-6-yl)L-lysine In a furnace-dried 250 mL Erlenmeyer flask, add 1-octyl β-D-glucuronic acid (Carbosynth Ltd., 3.06 g, 10 mmol), 50 mL of anhydrous DMF, and anhydrous 1-hydroxybenzotriazole (1.62 g, 12 mmol). Add the chilled (4°C) solution of N,N'-dicyclohexylcarbodiimide (2.48 g, 12 mmol) in 50 mL of DMF with stirring, and allow the reaction to proceed for 5 minutes. Filter the large amount of white precipitate of N,N'-dicyclohexylurea over a frit glass funnel, and add the filtrate to the solution of N-α-Fmoc-L-lysine (3.68 g, 10 mmol) in 25 mL of anhydrous DMF. Allow the reaction to proceed for 25 minutes with stirring, until it reaches room temperature or the ninhydrin color becomes very pale. The reaction mixture is filtered, removed and allowed to dry, then crystallized from MeOH / Et2O by dissolution in MeOH and slow dilution to the cloud point in Et2O, and subsequently by freezing. Further purification can be achieved by silica gel chromatography using a solvent gradient from ƒ to ƒ / EtOH / AcOH.

[0361] By a similar method, N-α-Boc-L-lysine can be substituted to obtain N-α-Boc,N-ε-(1-octylβ-D-glucuronide-6-yl)L-lysine, which is suitable for N-terminal incorporation and cleavage to the free N-terminus. By a similar method, N-α-Ac-L-lysine can be substituted to obtain N-α-AC,N-ε-(1-octylβ-D-glucuronide-6-yl)-L-lysine, which is suitable for N-terminal incorporation of peptides with a blocked N-terminus. By a similar method, an appropriate amount of N-α-Fmoc-L-ornithine can be substituted to obtain N-α-Fmoc,N-δ-(1-octylβ-D-glucuronide-6-yl)-L-ornithine. By a similar method, other N-mono-protected diamino acids can be substituted to obtain the corresponding reagents. Alternatively, the use of a transient Me3Si ester protecting group during coupling and without prior activation of 1-octyl β-D-glucuronic acid provides an easy route for reagent formation. The transient Me3Si ester is produced by the reaction of Fmoc-Lys-OH with equimolar amounts of N,O-bis(trimethylsilyl)acetamide in dichloromethane (CH2Cl2). The organic layer contains the desired reagent as a solution in CH2Cl2, ready for coupling with the 1-alkyl glucuronide as described above. The filtered reaction mixture is washed with aqueous NaHSO4 to hydrolyze the Me3Si ester, dried over MgSO4, and the solvent is removed.

[0362] Similarly, 1-octyl β-D-glucuronic acid of peracetyl or perbenzoyl is used to obtain Ac or Bz-protected forms of the reagent (e.g., 2,3,4-trisacetyl 1-octyl β-D-glucuronic acid formed by treatment with Ac2O). Such reagents have increased stability during acid cleavage from the resin and are used when instability during deprotection is detected (see Kihlberg, J., et al. (1997) Methods Enzymol 289: 221-245 and its references). Final deprotection of such products is carried out by base-catalyzed transesterification after cleavage using MeOH / NH3, MeOH / NaOMe, or MeOH / NH2NH2, as described above.

[0363] Example 2: Synthetic Peptide Analog Generally, peptide synthesis methods involve the sequential addition of protected amino acids to a growing peptide chain. Typically, either the amino group or carboxyl group and the reactive side chain group of the first amino acid are protected. This protected amino acid is then either bound to an inert solid support or utilized in solution, and the next amino acid in the sequence, appropriately protected, is added under conditions suitable for amide bond formation. After all desired amino acids have been linked in the appropriate sequence, the protecting groups and any solid support are removed to obtain a crude peptide. The peptide is desalted and purified by chromatography.

[0364] A preferred method for preparing analogues of physiologically active truncated peptides having fewer than approximately 50 amino acids involves solid-layer peptide synthesis. In this method, the α-amino(Nα) functional group and any reactive side chain are protected with an acid-sensitive or base-sensitive group. The protecting group should be stable to the peptide bond formation state while being easily removed without affecting the existing peptide chain. Suitable α-amino protecting groups include, but are not limited to, t-butyloxycarbonyl (Boc), benzyloxycarbonyl (Cbz), o-chlorobenzyloxycarbonyl, biphenylisopropyloxycarbonyl, t-amyloxycarbonyl (Amoc), isobornyloxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, o-nitrophenylsulfenyl, 2-cyano-t-butoxycarbonyl, and 9-fluorenyl-methoxycarbonyl (Fmoc) (preferably Boc, or more preferably Fmoc). Suitable side-chain protecting groups include, but are not limited to, acetyl, benzyl (Bzl), benzyloxymethyl (Bom), Boc, t-butyl, o-bromobenzyloxycarbonyl, t-butyl, t-butyldimethylsilyl, 2-chlorobenzyl (Cl-z), 2,6-dichlorobenzyl, cyclohexyl, cyclopentyl, isopropyl, pivaloyl, tetrahydropyran-2-yl, tosyl (Tos), 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf), trimethylsilyl, and trityl. The preferred Nα protecting group for the synthesis of the compounds is the Fmoc group. Preferred side-chain protecting groups are the Ot-butyl group for Glu, Tyr, Thr, Asp, and Ser; the Boc group for the side chains of Lys and Trp; the Pbf group for Arg; and the Trt group for Asn, Gln, and His. For selective modification of Lys residues, orthogonal protection is preferred, which has a protecting group that is not removed by Fmoc or t-butyl-based cleavage reagents.Preferred examples for modification of the Lys side chain include, but are not limited to, those removed by hydrazine rather than piperidine; for example, 1-(4,4-dimethyl-2,6-dioxocyclohexa-1-ylidene)-3-methylbutyl (ivDde) ​​or 1-(4,4-dimethyl-2,6-dioxocyclohexa-1-ylidene)ethyl (Dde) and allyloxycarbonyl (Alloc).

[0365] When side-chain lactam formation is desired, a scheme of Fmoc-Lys(ivDde) ​​or Fmoc-Lys(Dde) protecting groups is preferred (Houston, ME, Jr., et al. (1995) J Pept Sci 1: 274-282; Murage, EN, et al. (2010) J Med Chem), because in this case, Fmoc-Glu(O-allyl) and Fmoc-Lys(Alloc) can be incorporated and used to provide transient protection, which is then deprotected for lactam formation, while the Lys(Dde) protecting group remains for later removal and reaction with functionalized surfactants. The side-chain lactam between an acidic residue and a basic residue (e.g., Glu and Lys) is performed after removal of allyl-based protection by activation of the carboxyl side-chain functional group with N,N'-diisopropylcarbodiimide (DIC) / 1-hydroxybenzotriazole (HOBt) or 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium·hexafluorophosphate (HBTU) / N,N-di-isopropylethylamine (DIEA), using standard protocols well known in the art.

[0366] In solid-phase synthesis, the C-terminal amino acid is first bonded to a suitable resin support. Suitable resin supports are those that are insoluble in the culture medium used, in addition to being inert to the reagents and reaction conditions of the stepwise condensation and deprotection reactions. Examples of commercially available resins include styrene / divinylbenzene resins modified with reactive groups, e.g., chloromethylated co-poly-(styrene-divinylbenzene), hydroxymethylated co-poly-(styrene-divinylbenzene), etc. Benzylated, hydroxymethylated phenylacetamidomethyl (PAM) resins and hydroxymethylphenoxyacetylamidomethyl (HMPA) are preferred for the preparation of peptide C-terminal acids. When the C-terminus of the compound is an amide, preferred resins include p-methylbenzhydrylamino-co-poly(styrene-divinylbenzene) resins and 2,4-dimethoxybenzhydrylamino-based resins ("Rink amides"), etc. Particularly preferred supports for the synthesis of larger peptides are commercially available resins containing grafted PEG sequences on other polymer matrices, such as Rink Amide-PEG and PAL-PEG-PS resins (Applied Biosystems), or similar resins designed for peptide amide synthesis using the Fmoc protocol. Therefore, in certain cases, it is desirable to have an amide bond to the PEG chain. In these cases, it is preferable to bond an N-Fmoc-amino-PEG-carboxylic acid to the resin forming the amide (e.g., Rink amide resin). The first amino acid of the chain can be bonded to the amino functional group of the PEG chain as an N-Fmoc-amino acid. Final deprotection yields the desired peptide-NH-PEG-CO-NH2 product.

[0367] The bonding of PAM or HMPA to the resin can be achieved by reacting Nα-protected amino acids, such as Boc amino acids, with the resin at a high temperature, for example, between about 40°C and 60°C, preferably at about 50°C, for about 12 to 72 hours, preferably about 48 hours, using ammonium, cesium, triethylammonium, 1,5-diazabicyclo-[5.4.0]undec-5-ene, tetramethylammonium, or similar salts in ethanol, acetonitrile, N,N-dimethylformamide (DMF), preferably as cesium salts in DMF. This ultimately yields a peptide acid product after acid cleavage or an amide after aminolysis.

[0368] Nα-Boc-amino acids can be bonded to benzhydrylamine resins by (DIC) / HOBt-mediated coupling in a solvent such as CH2Cl2 or DMF, preferably CH2Cl2, at a temperature between about 10°C and 50°C, preferably 25°C, for about 2 hours to about 24 hours, preferably about 2 hours.

[0369] With regard to Boc-based protocols, sequential coupling of protected amino acids can typically be carried out in an automated peptide synthesizer by methods well known in the art. Each protected amino acid is introduced in a molar excess of approximately 1.5 to 2.5 times after neutralization with triethylamine, DIEA, N-methylmorpholine (NMM), colidine, or a similar base, and the coupling is carried out at ambient temperature in an inert, non-aqueous polar solvent, preferably dichloromethane, such as CH2Cl2, DMF, N-methylpyrrolidone (NMP), N,N-dimethylacetamide (DMA), or a mixture thereof. With regard to Fmoc-based protocols, no acid is used for deprotection, but a base (preferably DIEA or NMM) is usually incorporated into the coupling mixture. The coupling is typically carried out in DMF, NMP, DMA, or a mixed solvent, preferably DMF. Typical coupling agents are N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), or other carbodiimides, either alone or in the presence of HOBt, O-acylurea, benzotriazol-1-yl-oxytris(pyrrolidino)phosphonium hexafluorophosphate (PyBop), N-hydroxysuccinimide, other N-hydroxyimides, or oximes. Alternatively, protected amino acid active esters (e.g., p-nitrophenyl, pentafluorophenyl, etc.) or symmetric anhydrides may be used. Preferred coupling agents are aminium / uronium (alternative names used by suppliers) class coupling agents such as HBTU, O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU), and 2-(6-chloro-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate (HCTU).

[0370] A preferred method for binding to Fmoc-PAL-PEG-PS resin can be achieved by deprotecting the resin linker with 20% piperidine in DMF, followed by the reaction of the N-α-Fmoc-protected amino acid in a microwave-assisted peptide synthesizer with HBTU:di-isopropylethylamine (DIEA) (1:2) in DMF in a molar excess of approximately 5 times the amount of the N-α-Fmoc-amino acid, using HBTU:di-isopropylethylamine (DIEA) (1:2) in DMF, with a maximum coupling cycle of 75° for 5 minutes.

[0371] Certain analogs herein contain a dispersed PEG linker (dPEG) at the C-terminus. Such linkers are short chains of polyethylene having amino and carboxyl termini. Therefore, they are essentially non-natural amino acids and are treated for synthesis in the same way as other amino acids. For example, Fmoc-amidooxy-dPEG4-acid is commercially available from Quanta Biodesign (#10213) and is bonded to a Rink or HMPA resin in the first step of synthesis in a manner similar to that used for the N-Fmoc or N-Boc amino acids described above. Deprotection under standard strong acid conditions provides peptides modified at the C-terminus of the corresponding short dPEG having the corresponding acid or amide C-terminus.

[0372] For this Fmoc-based protocol in a microwave-assisted peptide synthesizer, the (N)-α-Fmoc amino acid protecting group is removed with 20% piperidine in DMF containing 0.1 M 1-hydroxybenzotriazole (HOBt) in a double deprotection protocol for 30 seconds and then 3 minutes at a temperature set to a maximum of 75°C. HOBt is added to the deprotection solution to reduce aspartimide formation. Subsequently, the coupling of the next amino acids is performed using HBTU:DIEA (1:2) with a 5-fold molar excess in a double coupling cycle at a maximum of 75°C for 5 minutes.

[0373] At the end of the solid-phase synthesis, the fully protected peptide is removed from the resin. When the binding to the resin support is of the benzyl ester type, cleavage can be achieved by aminolysis with an alkylamine or fluoroalkylamine for peptides having an alkylamide C-terminus, or by ammonia / methanol or ammonia / ethanol for peptides having an unsubstituted amide C-terminus, for example, for peptides having an unsubstituted amide C-terminus, at a temperature between about -10°C and 50°C, preferably about 25°C, for about 12 to 24 hours, preferably about 18 hours. Peptides having a hydroxy C-terminus may be cleaved by HF or other strongly acidic deprotection regimens, or by saponification. Alternatively, the peptide may be removed from the resin, for example, by transesterification with methanol, and then by aminolysis or saponification. The protected peptide can be purified by silica gel or reverse-phase HPLC.

[0374] Side chain protecting groups can be removed from the peptide by treating the aminolysis product with anhydrous liquid hydrogen fluoride in the presence of anisole or other carbonium ion scavengers, for example, with a hydrogen fluoride / pyridine complex, with tris(trifluoroacetyl)boron and trifluoroacetic acid, with hydrogen and palladium on carbon or polyvinylpyrrolidone, or with sodium in liquid ammonia, preferably liquid hydrogen fluoride and anisole, for a period of about 15 minutes to 2 hours, preferably about 1.5 hours, at a temperature between about -10°C and +10°C, preferably about 0°C.

[0375] With respect to peptides on benzhydrylamine-type resins, the resin cleavage and deprotection steps can be combined in a first step utilizing liquid hydrogen fluoride and anisole, either as described above or preferably via the use of milder cleavage cocktails. For example, with respect to PAL-PEG-PS resin, a preferred method is via the use of a double deprotection protocol in a microwave-assisted peptide synthesizer, using one of the mild cleavage cocktails known in the art, such as TFA / water / tri-isopropylsilane / 3,6-dioxa-1,8-octanedithiol (DODT) (92.5 / 2.5 / 2.5 / 2.5), for 18 minutes at 38°C per hour. Cleavage of alkyl glycoside-containing materials has shown the survival of alkyl glycoside bonds using protocols with TFA / water ratios ranging from 9 / 1 to 19 / 1. Typical cocktails are 94% TFA; 2% EDT; 2% H2O; 2% TIS. Typically, the completely deprotected product is precipitated, washed with cold (-70° to 4°) Et2O, dissolved in deionized water, and freeze-dried.

[0376] The peptide solution may be desalted (e.g., with BioRad AG-3® anion exchange resin), and the peptide may be purified by a series of chromatographic steps utilizing one or all of the following types: ion exchange on a weakly basic resin in acetate form; hydrophobic adsorption chromatography on non-derivativeized co-poly(styrene-divinylbenzene), e.g., Amberlite® XAD; silica gel adsorption chromatography; ion exchange chromatography on carboxymethylcellulose; partition chromatography on, e.g., Sephadex® G-25; countercurrent partitioning; supercritical fluid chromatography; or HPLC, in particular reversed-phase HPLC on octyl-bonded or octadecylsilylsilica (ODS)-bonded phase column packing.

[0377] This specification also provides a process for preparing covalently modified peptides and / or proteins and their pharmaceutically acceptable salts, the process comprising the steps of sequentially condensing protected amino acids on a suitable resin support, removing the protecting group and resin support, and purifying the product, thereby obtaining analogues of bioactive cleavage homologues and analogues of covalently modified peptides and / or proteins described herein. In some embodiments, the covalently modified peptides and / or proteins described herein incorporate alkylglycoside modifications as defined above. Another embodiment relates to a process for preparing covalently modified peptides and / or proteins and their pharmaceutically acceptable salts, the process comprising the use of a microwave-assisted solid-phase synthesis-based process or a standard peptide synthesis protocol for sequentially condensing protected amino acids on a resin support, as defined above, removing the protecting group and resin support, and purifying the product, thereby obtaining analogues of bioactive peptides.

[0378] Example 3. General oxidation method for uronic acid To a solution of 1-dodecyl β-D-glucopyranoside (Carbosynth) [2.0 g, 5.74 mmol] in 20 mL of acetonitrile and 20 mL of deionized (DI) water, (diacetoxyiodo)benzene (Fluka) [4.4 g, 13.7 mmol] and TEMPO (Sigma-Aldrich) [0.180 g, 1.15 mmol] were added. The resulting mixture was stirred at room temperature for 20 hours. After the reaction, mass spectrometry (e.g., LCQ ESI) was performed. Upon completion, the reaction mixture was diluted with water, lyophilized, and yielded 1.52 g (crude yield 73.1%) of the crude product, 1-dodecyl β-D-glucuronic acid, as a white powder, which was used directly in solid-phase synthesis without further purification. This product was also used for longer alkyl groups by an alternative process using NaOCl as an oxidant, as described herein. For longer alkyl groups, 1,4-dioxane was used instead of acetonitrile, and the temperature was increased to 30°C. Similar methods were used to prepare the desired alkylsaccharide uronic acids used to produce the products and reagents described herein.

[0379] Similar methods were used, for example, to prepare the desired 1-alkylsaccharide uronic acid using the corresponding 1-octyl, 1-decyl, 1-undecyl, 1-tetradecyl, 1-hexadecyl, and 1-octadecyl glycosides (purchased from Anatrace, Maumee, OH), which were then used to produce the products and reagents described herein. Similarly, for example, the desired 1-alkyl disaccharide uronic acid was prepared using the corresponding 1-octyl, 1-decyl, 1-undecyl, 1-tetradecyl, 1-hexadecyl, and 1-octadecyl β-D-meribioside or β-D-maltoside (purchased from Anatrace, Maumee, OH), which were then used to produce the products and reagents described herein.

[0380] Example 4: Preparation of C-terminal amide analog (EU-A387). Samples of the Fmoc-His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Bip-Ser-Lys-Tyr-Leu-Glu-Ser-Lys(Alloc)-Rink amide resin were prepared by sequential addition of N-α-Fmoc-protected amino acids as described in Example 1, and deprotected at the Lys-N-epsilon position by incubation overnight at room temperature in the dark with Pd(PPh3)4 (0.5 equivalents) and DMBA (20 equivalents) in DMF / CH2Cl2 (1:1). After washing with DMF / CH2Cl2, the Lys side chain was acylated with 1'-dodecyl β-D-glucuronic acid in DMF / CH2Cl2 via the use of DIC / HOBt. Completion of coupling was confirmed by ninhydrin, and the product was broadly washed with CH2Cl2.

[0381] The product resin was subjected to final deprotection and cleavage by treatment with a cleavage cocktail (94% TFA: 2% EDT; 2% H2O; 2% TIS) at room temperature for 240 minutes. The mixture was treated with Et2O to precipitate the product, and after extensive washing with Et2O and drying in vacuum, the crude title peptide product was obtained.

[0382] Purification was performed in two batches by reverse-phase (C18) HPLC. The crude peptide was packed onto a 4.1 x 25 cm HPLC column at a flow rate of 15 mL / min (15% organic modifier; acetate buffer) and eluted at 50°C for 60 minutes with a gradient from 15-45% buffer B. The fraction of the product was lyophilized and analytical HPLC (18.6 min; 30-60% CH3CN in 0.1% TFA) / mass spectrometry (M+1 peak = 2382.14) yielded the title product peptide with a purity of 98.03%. Other analogs of the present invention were prepared by a similar method, and their characteristics are illustrated below. The corresponding 1-methyl and 1-octyl analogs of the title compound were prepared by a similar method, but using the reagents 1'-methyl β-D-glucuronic acid and 1'-octyl β-D-glucuronic acid (Carbosynth). The corresponding 1-decyl, 1-dodecyl, 1-tetradecyl, 1-hexadecyl, 1-octadecyl, and 1-eicosyl and higher analogs are prepared using the corresponding monosaccharide and disaccharide uronic acids prepared as described above. Alternatively, 1-alkylglucuronyl or other uronic acid acylated analogs may be prepared by initial purification of a deprotected or partially deprotected peptide followed by acylation with a desired uronic acid reagent.

[0383] Example 5: Preparation of peptide C-terminal acid analogs. Samples of Boc-His(Trt)-Aib-Gln(Trt)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Lys(Boc)-Tyr(tBu)-Leu-Asp(OtBu)-Glu(O-allyl)-Gln(Trt)-Ala-Ala-Lys(Alloc)-Glu(O-tBu)-Phe-Ile-Lys(Dde)-Trp(Boc)-Leu-Leu-Gln(Trt)-Thr(tBu)-HMPA, resin (Fmoc-Thr(tBu)-HMPA)Resin, starting at 0.45 mmol / g substitution, were prepared by sequential addition of N-α-Fmoc-protected amino acids as described in Example 1. The allyl-based side chains on Glu and Lys were deprotected by incubation in DMF / CH2Cl2 (1:1) with Pd(PPh3)4 (0.5 equivalents) and DMBA (20 equivalents) overnight at room temperature in the dark. The resin was washed with 0.5% DIEA in DMF (twice), 0.5% sodium diethyldithiocarbamate in DMF (twice), and DMF / CH2Cl2 to obtain a pale yellow resin. Side-chain lactam bonds were formed by bonding Glu and Lys with DIC / HOBT (5 equivalents) in DMF. The reaction was checked for integrity with ninhydrin, and recombination was performed as necessary. After washing with DMF / CH2Cl2, the Lys side chain was deprotected twice by incubation in DMF (10 equivalents) with 5% hydrazine hydrate for 15 minutes each time. After washing with DMF / CH2Cl2, the side-chain amino groups of the deprotected Lys residues were reacted with 1'-tetradecyl β-D-melibiouronic acid in DMF / CH2Cl2 via DIC / HOBt. Completion of coupling was confirmed with ninhydrin, and the product was broadly washed with CH2Cl2. Coupling that was not complete was re-executed with ninhydrin. Generally, 10-12 g of peptide product resin was obtained from the synthesis of 2 mmol.

[0384] The product resin was subjected to final deprotection and cleavage by treatment with a cleavage cocktail (94% TFA: 2% EDT; 2% H2O; 2% TIS) at room temperature for 240 minutes. The mixture was treated with Et2O to precipitate the product, and after extensive washing with Et2O and drying in vacuum, the crude title peptide product was obtained. Generally, 5-8 g of crude product peptide was obtained.

[0385] Purification was performed in two batches by reverse-phase (C18) HPLC. The crude peptide was packed onto a 4.1 x 25 cm HPLC column at a flow rate of 15 mL / min (15% organic modifier; 0.1% TFA buffer) and eluted at room temperature for 70 minutes with a gradient from 35-55% buffer B. The fraction with lower purity was re-purified for the fraction having a purity of >70%. The fraction of the product was lyophilized and analytical HPLC (10.3 min; 45-75% CH3CN in 0.1% TFA) / mass spectrometry (1317.67, +3 packing; 1976.13, +2 packing; molecular weight 3950.44) yielded EU-A1077 with a purity of 98.7%. Other analogues of the present invention were prepared by similar methods, and their characteristics are illustrated below.

[0386] Corresponding 1-methyl and 1-octyl analogs of the title compound are prepared by a similar method using the reagents 1'-methyl β-D-glucuronic acid and 1'-octyl β-D-glucuronic acid (Carbosynth). The products of the present invention were prepared by a similar method using the corresponding β-D-glucuronic acid of 1-octyl, 1-decyl, 1-undecyl, 1-tetradecyl, 1-hexadecyl, and 1-octadecyl (prepared as described above). The products and reagents described herein were prepared by a similar method using the corresponding β-D-melibiouronic acid or β-D-maltouronic acid of 1-octyl, 1-decyl, 1-undecyl, 1-tetradecyl, 1-hexadecyl, and 1-octadecyl (prepared as described above). Alternatively, 1-alkylglucuronyl or other uronic acid acylated analogs may be prepared by initial purification of a deprotected or partially deprotected peptide followed by acylation with a desired uronic acid reagent. Alternatively, ammonolisis of an HMPA resin-bound intermediate yields the corresponding amide C-terminus.

[0387] Analysis and characterization were performed by HPLC / mass spectrometry in cationic mode using the elution gradient given in the table below.

[0388] [Table 1-1]

[0389] [Table 1-2]

[0390] The additional compounds synthesized and analyzed as described above are as follows:

[0391] [Table 2-1]

[0392] [Table 2-2]

[0393] [Table 2-3]

[0394] [Table 2-4]

[0395] Example 6: Compound cell assay. The compound was accurately weighed to approximately 1 mg and analyzed using a standard cell assay (Cerep SA). The readout was the amount of cAMP produced in cells treated with the test compound, either as an agonist or antagonist. The assays used were stimulation of cAMP levels during cell assays of glucagon (human, cloned into CHO cells) and GLP-1 (mouse cell line). The assays are described in Chicchi, GG, et al. (1997) J Biol Chem 272: 7765-7769 and Runge, S., et al. (2003) Br J Pharmacol 138: 787-794.

[0396] Regarding compound EU-A391, the GLCR cell response remained unchanged, while the GLP1R cell response was significantly elevated at an EC50 of 420 nM.

[0397] [Table 3]

[0398] A further series of cell assays were performed using standard cell assays (DiscoveRx, LeadHunter assays) employing cAMP stimulation or arrestin activation readout. Compounds were accurately weighed in approximately 1 mg quantities and sent to the DiscoveRx for dilution and assay. The assays used were for the receptors of glucagon (cloned into human CHO cells) and GLP-1 (cloned into human CHO cells) in the cell assays.

[0399] [Table 4-1]

[0400] [Table 4-2]

[0401] [Table 4-3]

[0402] Example 7: In vivo assay of compounds - db / db mice Sixty female db / db B6BKS(D)Leprdb / J (strain 000697) mice used in this study were approximately 8–9 weeks old upon arrival (Jackson Laboratory, Bar Harbor, Maine). The mice were randomized by body weight into two treatment groups of eight female mice each, administered the test substance, EU-A994, EU-A995, or EU-A1026, at dose levels of 100 nmoles / kg or 300 nmoles / kg. One group of eight female mice served as a vehicle control and received the vehicle, 0.2% BSA, in saline, pH 7.4. An additional group of eight female mice received the positive control substance, liraglutide, at a dose level of 50 nmoles / kg. The test substance, vehicle, and positive control substance were administered by subcutaneous injection at a dose of 6 mL / kg during approximately hours 0, 7, and 24 of day 1 of the study.

[0403] Clinical observations were performed before randomization and daily from day 1 to day 5 at the time of receipt. Body weight was measured and recorded before randomization and daily from day 1 to day 5 at the time of receipt. Food intake was measured and recorded daily from day 1 to day 5. Blood samples for glucose analysis were collected before the study (-3 days) and at 0, 1, 2, 4, 8, 10, 24, 48, 72, and 96 hours after the first dose on day 1. At the end of the study, all animals were euthanized and the carcasses were discarded without further evaluation.

[0404] Significant weight changes were observed on days 2 and 3 for the vehicle and high doses of EU-A994 and high doses of EU-A1026 for liraglutide, and on days 3 and 4 for low doses of EU-A1026. In the analysis of food intake, animals treated with liraglutide differed significantly from those treated with the vehicle on days 1 and 2, high doses of EU-A994 on day 1, and low doses of EU-A995 on days 1 and 2. High doses of EU-A994 on day 1, and high and low doses of EU-A1026 on day 2 differed significantly from those treated with liraglutide. Glucose levels for liraglutide at 10 hours and for high doses of EU-A994 at 10 and 24 hours differed significantly from those treated with the vehicle. Low doses of EU-A995 and EU-A1026 at 10 hours differed significantly from those treated with liraglutide (Figure 5). In a similar manner, other analogs from the same series were tested for their effects on blood glucose, body weight, and food intake.

[0405] [Table 5]

[0406] Example 8: In vivo assay of compounds - DIO mouse Fifty (50) male C57BL / 6J mice, classified as diet-induced obesity (DIO), were received from the JAXA Institute at six weeks of age. The mice's ears were clipped for identification, and they were individually housed in polycarbonate cages with positive pressure ventilation using HEPA-filtered air, at a density of five mice per cage. The animal room was illuminated with artificial fluorescent lighting under a 12-hour light / dark cycle. The standard temperature and relative humidity ranges in the animal room were 22±4°C and 50±15%, respectively. Filtered tap water, acidified to a pH of 2.8 to 3.1, was provided as a free-flowing high-fat diet (Research Diets D12492; 60 kcal %).

[0407] After a two-week acclimatization period, 50 mice were randomized into the following groups (n=10): Group 1: Vehicle-treated; Group 2: Low-dose EU-A994; Group 3: High-dose EU-A594; Group 4: Low-dose EU-A1024; Group 5: High-dose EU-A1024. Subcutaneous injection (SC) was administered to the mice on days 1 (0, 7 hours), 3, 6, 9, 12, 15, 18, 21, and 24. Body weight and lateral observations of the cage were recorded daily. Food and water intake were recorded weekly. Mice underwent NMR measurements on day 1 (pre-administration) and day 26 to determine whole-body fat and lean components. On day 26, the mice were fasted overnight for an oral glucose tolerance test. The following day, the first blood sample was collected via tail nick (t=0). Subsequently, mice were administered a bolus of 1.0 g / kg glucose. Blood samples were obtained via tail nicks at 0, 15, 30, 60, 90, and 120 minutes, with glucose and plasma glucose immediately measured using a glucometer.

[0408] Sacrifice and tissue collection: Mice were sacrificed on day 28. Terminal blood was processed into serum / plasma, and fractions were sent for analysis of glucose, insulin, and lipid profiles. Body composition was determined by NMR. Typical compound EU-A1024 showed decreased glucose excursion during OGTT, increased glucose-dependent insulin secretion, decreased basal insulin secretion, decreased body weight gain (Figure 10), and decreased body fat mass, but had the least effect on lean body mass (Figure 11).

[0409] Example 9: Plasma protease stability Outline of the development of the bioanalytical method: I. Equipment used; API-4000 mass spectrometer, ESI positive, MRM scan; Shimadzu HPLC / CTC Autosampler with ACE C8 column (2.1 x 50 mm, 5 μm), mobile phase A: 0.1% formic acid, 5 mM NH4OAc in water, mobile phase B: 0.1% formic acid in CH3CN, 10 μL of sample injected; II. Standard and QC sample preparation: i. 1 mg / mL storage solution was prepared in DMSO / CH3CN (1 / 1); ii. Standard working solutions were prepared by diluting the storage solution in 50% CH3CN. The concentrations of the diluted standard solutions were 20, 50, 100, 200, 500, 1000, 2000, 5000, 10000, and 20000 ng / mL; iii. Add 10 μL of diluted standard solution to 90 μL of blank plasma and mix them; iv. Add 300 μL of internal standard solution (verapamil, 20 ng / mL in 100% CH3CN), vortex, and centrifuge; v. Transfer the supernatant to an HPLC injection plate for packing onto the HPLC column; vi. The standard samples were 2, 5, 10, 20, 50, 100, 200, 500, 1000, and 2000 ng / mL. The QC samples were 5 (LQC), 50 (MQC), and 500 (HQC) ng / mL. For plasma stability studies, samples of EU-A993, EU-A1023, and human GLP-1 (7-36, Bachem) were prepared in human plasma (approximately 6-20 ng / mL or similar concentrations exceeding the limit of quantification) and sampled at time points t=0, 0.5, 1, 2, 4, and 8 hours during incubation at 30°C. Samples were treated with an internal standard solution (100% CH3CN) as described above to precipitate proteins, and the supernatant (supernatentants) was packed into injection plates and loaded onto an HPLC column for quantification by mass spectrometry. Plots of the compound signal (amount) versus time showed a rapid decrease for GLP-1 (7-36), while the amount of the intact compound in human plasma for the compounds of the present invention remained essentially unchanged over 8 hours (see Figure 8).

[0410] Example 10: Use of the compound. The covalently modified peptides and / or proteins described herein are useful for the prevention and treatment of various diseases associated with obesity, metabolic syndrome, cardiovascular disease, and diabetes. Peptides modified with appropriately labeled surfactants can be used as diagnostic probes.

[0411] Typical delivery regimens include oral, parenteral (including subcutaneous, intramuscular, and intravenous injection), rectal, buccal (including sublingual), transdermal, inhalation, ophthalmic, and intranasal administration. An attractive and widely used method for peptide delivery involves subcutaneous injection of controlled-release injectable formulations. Other routes of administration for the application of covalently modified peptides and / or proteins described herein are subcutaneous, intranasal, and inhalation administration.

[0412] Example 11. Pharmaceutical use for the treatment of insulin resistance. Human patients with evidence of insulin resistance or metabolic syndrome are treated with EU-A596 by intranasal administration (200 μL) from a standard atomizer used in the art, containing a solution of the drug in saline solution containing 0.5 mg / mL to 10 mg / mL of the drug and standard excipients such as benzyl alcohol. This treatment is repeated as needed to alleviate symptoms such as obesity and elevated blood glucose. In a similar manner, a solution of EU-A596 and selected excipients in an evaporative solvent containing hydrofluoroalkanes, etc., is administered intranasally by a medium-dose inhaler (MDI) as needed to reduce insulin resistance. The therapeutic effect is assessed using standard tests, including measurement of blood glucose levels, body mass index, and / or body weight, and / or waist-to-hip ratio.

[0413] In a similar manner, adjusted doses administered via transchucine, intravaginal, inhalation, subcutaneous, intravenous, intraocular, or oral routes are tested to determine the level of GLP1R and / or GLCR stimulation on cells throughout the body and to assess therapeutic effects.

[0414] <array> This specification provides sequences of SEQ.ID.No.1-3 and SEQ.ID.No.774-783, 785-797, and 1025-1029. Furthermore, Table 1 in Figure 1 provides sequence numbers for compounds EU-A300 to EU-A425, each having SEQ.ID.No.4-129. The compounds and their respective sequence numbers in Table 1 in Figure 1 are incorporated into the specification of this application. Furthermore, Table 2 in Figure 2 provides sequence numbers for compounds EU-A426 to EU-A599, each having SEQ.ID.No.130-317. The compounds and their respective sequence numbers in Table 2 in Figure 2 are incorporated into the specification of this application. Furthermore, Table 3 in Figure 3 provides sequence numbers for compounds EU-A700 to EU-A1174, each having SEQ.ID.No.318-773;798-806. The compounds and their respective sequence numbers in Table 3 of Figure 3 are incorporated into the specification of this application. Furthermore, Table 4 of Figure 9 provides sequence numbers for compounds EU-A1575 to EU-A1870, having SEQ.ID.No. 807-1024 and 1030-1107, respectively. The compounds and their respective sequence numbers in Table 4 of Figure 9 are incorporated into the specification of this application.

Claims

1. A peptide product comprising a surfactant X covalently bonded to the peptide, The peptide contains the linker amino acid U and at least one other amino acid: 【Chemistry 1】 In the formula, surfactant X is the group of formula I: 【Chemistry 2】 During the ceremony: R １ａ Independently, at each occurrence, a single bond, H, protecting group, saccharide, substituted or unsubstituted C １ -C ３０ The moiety contains an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid core. R １ｂ , R １ｃ , and R １ｄ Each of these elements, independently, appears as a protecting group, saccharide, single bond, H, or substituted or unsubstituted C at each instance. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, W １ is independently, at each occurrence, -CH ２ -, -CH ２ -O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH ２ -S-, and W ２ ha-O-, -CH ２ -, or -S-, R ２ Independently, at each occurrence, U is a single bond, H, substitution, or unsubstituted C. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, -NH, -S-, -triazolo-, -NH(C=O)-CH ２ -, - (CH ２ ) ｍ - It is maleimide, n is 1, 2, or 3, and, m is an integer between 1 and 10. The peptide was selected from formula II: aa １ -aa ２ -aa ３ -aa ４ -aa ５ -aa ６ -aa ７ -aa ８ -aa ９ -aa １０ -aa １１ -aa １２ -aa １３ -aa １４ -aa １５ -aa １６ -aa １７ -aa １８ -aa １９ -aa ２０ -aa ２１ -aa ２２ -aa ２３ -aa ２４ -aa ２５ -aa ２６ -aa ２７ -aa ２８ -aa ２９ -aa ３０ -aa ３１ -aa ３２ -aa ３３ -aa ３４ -aa ３５ -aa ３６ -aa ３７ -Z Formula II (SEQ. ID. NO. 1) During the ceremony: Z is OH, N-R ４ -His, or -NH-R ３ And, Here, R ３ H, substituted or unsubstituted C １ -C １２ Alkyl or PEG chains less than 10 Da, R ４ is C ２ -C １０ An acyl group, for example, Ac or Bz. aa １ His, N-R ４ -His, pGlu-His, or N-R ３ -His, aa ２ These are Ser, D-Ser, Ala, Gly, Pro, MePro, Aib, Ac4c, or Ac5c. aa ３ is Gln or Cit, aa ４ It is Gly or D-Ala, aa ５ is either Thr or Ser, aa ６ is Phe, Trp, 2FPhe, MePhe, 2FMePhe, or Naal2, aa ７ is either Thr or Ser, aa ８ is either Ser or Asp, aa ９ is Asp or Glu, aa １０ is Tyr, Leu, Met, Nal2, Bip, Bip2EtMeO, or U, aa １１ It does not exist, or it is Ser, Asn, Bip, or U. aa １２ It does not exist, or it is Lys, Glu, Ser, Arg, or U. aa １３ It does not exist, or it is Tyr, Glun, Cit, or U. aa １４ It does not exist, or it is Leu, Met, Nle, or U. aa １５ It does not exist, or it is Asp, Glu, or U. aa １６ It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Ac5c, Lys, Arg, or U. aa １７ It does not exist, or it is Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c, Lys, or U. aa １８ It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U. aa １９ It does not exist, or it is Ala, Val, Aib, Ac4c, Ac5c, or U. aa ２０ It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U. aa ２１ It does not exist, or it is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U. aa ２２ It does not exist, or it is Phe, Trp, Nal2, Aib, Ac4c, Ac5c, or U. aa ２３ is absent, or is Val, Ile, Aib, Ac4c, Ac5c, or U, aa ２４ It does not exist, or it is Ala, Gln, Glu, Cit, or U. aa ２５ It does not exist, or it is Trp, Nal2, or U. aa ２６ It does not exist, or it is Leu, or U. aa ２７ It does not exist, or it is Met, Val, Leu, Nle, Lys, or U. aa ２８ It does not exist, or it is Asn, Lys, Glun, Glu, Cit, or U. aa ２９ It does not exist, or it is Thr, Gly, Aib, Ac4c, Ac5c, or U. aa ３０ is absent, or is Lys, Aib, Ac4c, Ac5c, Arg, or U, aa ３１ It does not exist, or it is Arg, Aib, Ac4c, Ac5c, or U. aa ３２ It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U. aa ３３ It does not exist, or it is Arg, Aib, Ac4c, Ac5c, or U. aa ３４ It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U. aa ３５ It does not exist, or it is Asn, Aib, Ac4c, Ac5c, or U. aa ３６ It does not exist, or it is Ile, Aib, Ac4c, Ac5C, or U. aa ３６ It does not exist, or it is Ala, Aib, Ac4c, Ac5C, or U. aa ３７ It does not exist, or it is U. U is a natural or non-natural amino acid containing a functional group used for covalent bonding to surfactant X. Here, aa １ -aa ３７ Any two of them can be optionally cyclized via their side chains to form a lactam bond. However, aa １０ -aa ３７ A peptide product, assuming that one or at least one of them is a linker amino acid U covalently bonded to X.

2. The peptide product according to claim 1, wherein n is 1 or 2.

3. X has the following structure: 【Transformation 3】 During the ceremony: R １ａ This includes H, protecting groups, saccharides, and substituted or unsubstituted C. １ -C ３０ This is a portion containing an alkyl group or a steroid nucleus. R １ｂ , R １ｃ , and R １ｄ Each of these elements, independently, appears as H, a protecting group, a saccharide, or a substituted or unsubstituted C at each instance. １ -C ３０ It is an alkyl group, W １ These are independent, and at each occurrence, -CH ２ -ien-CH ２ -O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH ２ -S-, W ２ is -O-, -S-, R ２ is a single bond, -NH-, -S-, -NH(C=O)-CH ２ - or - (CH ２ ) ｍ - Maleimide - and, The peptide product according to claim 1, wherein m is an integer between 1 and 10.

4. The peptide product according to claim 3, wherein X has the following structure. 【Chemistry 4】

5. The peptide product according to claim 3, wherein X has the following structure. 【Transformation 5】

6. X has the following structure: 【Transformation 6】 During the ceremony: R １ａ This includes H, protecting groups, saccharides, and substituted or unsubstituted C. １ -C ３０ This is a portion containing an alkyl group or a steroid nucleus. R １ｂ , R １ｃ , and R １ｄ Each instance independently involves H, a protecting group, or a substituted or unsubstituted C. １ -C ３０ It is an alkyl group, W １ is -(C=O)-NH-, W ２ is -O-, R ２ The peptide product according to claim 3, wherein the bond is a single bond.

7. X has the following structure: 【Transformation 7】 During the ceremony: R １ａ C is either substituted or non-substituted. １ -C ３０ It is an alkyl group, R １ｂ , R １ｃ , and R １ｄ H is, W １ is -(C=O)-NH-, W ２ is -O-, and, R ２ The peptide product according to claim 3, wherein the bond is a single bond.

8. R １ａ C is either substituted or non-substituted. １ -C ３０ The peptide product according to claim 1 or 3, wherein the alkyl group is an alkyl group.

9. R １ａ C is either substituted or non-substituted. ６ -C ２０ The peptide product according to claim 1 or 3, wherein the alkyl group is an alkyl group.

10. R １ａ C is either substituted or non-substituted. １２ -C ２０ The peptide product according to claim 1 or 3, wherein the alkyl group is an alkyl group.

11. The peptide product according to claim 1 or 3, wherein surfactant X is a surfactant of the 1-alkyl glycoside class.

12. X is 1-eicosyl beta-D-glucuronic acid, 1-octadecyl beta-D-glucuronic acid, 1-hexadecyl beta-D-glucuronic acid, 1-tetradecyl beta-D-glucuronic acid, 1-dodecyl beta-D-glucuronic acid, 1-decyl beta-D-glucuronic acid, 1-octyl beta-D-glucuronic acid, 1-eicosyl beta-D-diglucuronic acid, 1-octadecyl beta-D-diglucuronic acid, 1- Xadecylbeta-D-diglucuronic acid, 1-tetradecylbeta-D-diglucuronic acid, 1-dodecylbeta-D-diglucuronic acid, 1-decylbeta-D-diglucuronic acid, 1-octylbeta-D-diglucuronic acid, or functionalized 1-eicosylbeta-D-glucose, 1-octadecylbeta-D-glucose, 1-hexadecylbeta-D-glucose, 1-tetradecylbeta-D-glucose, 1-do Decylbeta-D-glucose, 1-decylbeta-D-glucose, 1-octylbeta-D-glucose, 1-eicosylbeta-D-maltoside, 1-octadecylbeta-D-maltoside, 1-hexadecylbeta-D-maltoside, 1-tetradecylbeta-D-maltoside, 1-dodecylbeta-D-maltoside, 1-decylbeta-D-maltoside, or 1-octylbeta-D-maltoside, 1- The peptide product according to claim 1 or 3, comprising eicosylbeta-D-melibioside, 1-octadecylbeta-D-melibioside, 1-hexadecylbeta-D-melibioside, 1-tetradecylbeta-D-melibioside, 1-dodecylbeta-D-melibioside, 1-decylbeta-D-melibioside, or 1-octylbeta-D-melibioside, and the corresponding 6 or 6,6'-dicarboxylate salt.

13. The peptide product according to any one of claims 1 to 12, wherein U is selected from Lys, Cys, Orn, Glu, or a non-natural amino acid containing a functional group used for covalent bonding to surfactant X.

14. It has the structure of formula III-A, aa １ -aa ２ -aa ３ -aa ４ -aa ５ -aa ６ -aa ７ -aa ８ -aa ９ -aa １０ -aa １１ -aa １２ -aa １３ -aa １４ -aa １５ -aa １６ -aa １７ -aa １８ -aa １９ -aa ２０ - aa ２１ -aa ２２ -aa ２３ -aa ２４ -aa ２５ -aa ２６ -aa ２７ -aa ２８ -aa ２９ -Z Formula III-A (SEQ. ID. NO. 2) During the ceremony: Z is OH or -NH-R ３ And, R ３ H, substituted or unsubstituted C １ -C １２ Alkyl or PEG chains less than 10 Da, aa １ This refers to His, N-Ac-His, pGlu-His, or N-R ３ -His, aa ２ These are Ser, Ala, Gly, MePro, Aib, Ac4c, or Ac5c. aa ３ is Gln or Cit, aa ４ It is Gly or D-Ala, aa ５ is either Thr or Ser, aa ６ is Phe, Trp, 2FPhe, MePhe, 2FMePhe, or Naal2, aa ７ is either Thr or Ser, aa ８ is either Ser or Asp, aa ９ is Asp or Glu, aa １０ is Tyr, Leu, Met, Nal2, Bip, Bip2EtMeO, or U(X), aa １１ It does not exist, or it is Ser, Asn, Bip, or U(X), aa １２ It does not exist, or it is Lys, Glu, Ser, Arg, or U(X), aa １３ It does not exist, or it is Tyr, Glun, Cit, or U(X), aa １４ It does not exist, or it is Leu, Met, Nle, or U(X), aa １５ It does not exist, or it is Asp, Glu, or U(X), aa １６ It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Ac5c, Lys, Arg, or U(X), aa １７ It does not exist, or it is Arg, hArg, Gln, Glu, Lys, Cit, Aib, Ac4c, Ac5c, or U(X), aa １８ It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X), aa １９ It does not exist, or it is Ala, Val, Aib, Ac4c, Ac5c, or U(X), aa ２０ It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U(X), aa ２１ It does not exist, or it is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U(X), aa ２２ It does not exist, or it is Phe, Trp, Nal2, Aib, Ac4c, Ac5c, or U(X), aa ２３ It does not exist, or it is Val, Ile, Aib, Ac4c, Ac5c, or U(X), aa ２４ It does not exist, or it is Ala, Gln, Glu, Cit, or U(X), aa ２５ It does not exist, or it is Trp, Nal2, or U(X), aa ２６ It does not exist, or it is Leu, or U(X), aa ２７ It does not exist, or it is Met, Val, Leu, Nle, Lys, or U(X), aa ２８ It does not exist, or it is Asn, Lys, Glun, Glu, or U(X), aa ２９ It does not exist, or it is Thr, Gly, Aib, Ac4c, Ac5c, or U(X), Here, aa １ -aa ２９ Any two of them can be optionally cyclized via side chains to form a lactam bond. However, aa １０ aa １１ aa １２ aa １６ aa １７ aa １８ aa １９ aa ２０ aa ２１ aa ２２ aa ２３ aa ２４ aa ２５ aa ２６ aa ２７ aa ２８ , or aa ２９ The peptide product according to claim 1 or 3, wherein one or at least one of the is a natural or unnatural amino acid U covalently bonded to X.

15. It has the structure of formula III-B, His １ -aa ２ -aa ３ -Gly ４ -Thr ５ -aa ６ -Thr ７ -Ser ８ -Asp ９ -aa １０ -aa １１ -aa １２ -aa １３ -aa １４ -aa １５ -aa １６ -aa １７ -aa １８ -aa １９ -aa ２０ -aa ２１ -aa ２２ -aa ２３ -aa ２４ -aa ２５ -aa ２６ -aa ２７ -aa ２８ -aa ２９ -aa ３０ -Z Formula III-B (SEQ. ID. NO. 3) During the ceremony: Z is OH or -NH-R ３ And, R ３ H, substituted or unsubstituted C １ -C １２ Alkyl or PEG chains less than 10 Da, aa ２ It is Gly, MePro, or Aib, aa ３ is Gln or Cit, aa ６ is Phe, 2FPhe, MePhe, 2FMePhe, or Naal2, aa １０ is Tyr, Nal2, Bip, Bip2EtMeO, or U(X), aa １１ It does not exist, or it is Ser, Asn, Bip, or U(X), aa １２ It does not exist, or it is Lys, Glu, Ser, or U(X), aa １３ It does not exist, or it is Tyr, Glun, Cit, or U(X), aa １４ It does not exist, or it is Leu, Nle, or U(X), aa １５ It does not exist, or it is Asp, Glu, or U(X), aa １６ It does not exist, or it is Ser, Gly, Glu, Ala, Aib, Lys, Arg, or U(X), aa １７ It does not exist, or it is Arg, hArg, Gln, Glu, Lys, Cit, Aib, or U(X), aa １８ It does not exist, or it is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X), aa １９ It does not exist, or it is Ala, Aib, or U(X), aa ２０ It does not exist, or it is Gln, Lys, Arg, Cit, Glu, Aib, or U(X), aa ２１ It does not exist, or it is Asp, Glu, Leu, Aib, or U(X), aa ２２ It does not exist, or it is Phe, or U(X), aa ２３ It does not exist, or it is Val, Ile, Aib, or U(X), aa ２４ It does not exist, or it is Ala, Gln, or U(X), aa ２５ It does not exist, or it is Trp, or U(X), aa ２６ It does not exist, or it is Leu, or U(X), aa ２７ It does not exist, or it is Met, Val, Leu, Nle, Lys, or U(X), aa ２８ It does not exist, or it is Asn, Gln, Cit, or U(X), aa ２９ It does not exist, or it is Thr, Aib, or U(X), aa ３０ It does not exist, or it is Arg, or U(X), Here, aa １ -aa ２３ Any two of them can be optionally cyclized by their side chains to form a lactam bond. However, aa １０ aa １１ aa １２ aa １６ aa １７ aa １８ aa １９ aa ２０ aa ２１ aa ２２ aa ２３ aa ２４ , or aa ２８ The peptide product according to claim 1 or 3, wherein one or at least one of the is a natural or non-natural amino acid U covalently bonded to X.

16. aa １７ The peptide product according to claim 1, 14, or 15, wherein is a lysine residue bound to X.

17. aa ２０ The peptide product according to claim 1, 14, or 15, wherein is a lysine residue bound to X.

18. aa ２４ The peptide product according to claim 1, 14, or 15, wherein is a lysine residue bound to X.

19. aa ２８ The peptide product according to claim 1, 14, or 15, wherein is a lysine residue bound to X.

20. aa ２ The peptide product according to claim 1, 14, or 15, wherein is a glycine residue.

21. aa ２ The peptide product according to claim 1, 14, or 15, wherein is an Aib residue.

22. The peptide product according to claim 1, 14, or 15, wherein the peptide comprises one or more Aib residues.

23. It has the following structure: His １ -aa ２ -Gln ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ -Asp ９ -Tyr １０ -Ser １１ -Lys １２ -Tyr １３ -Leu １４ -Asp １５ -aa １６ -aa １７ -Ala １８ -Ala １９ -aa ２０ -Glu ２１ -Phe ２２ -aa ２３ -aa ２４ -Trp ２５ -Leu ２６ -aa ２７ -aa ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.774) During the ceremony, aa ２ is Gly or Aib, aa １６ is Glu, Ser, Ala, Lys, or Aib, aa １７ is Gln, Lys, or U(X), aa ２０ is Lys, Glu, or Arg, aa ２３ is Ile or Val, aa ２４ is Ala, Gln, or U(X), aa ２７ It is Met, Val, or Leu, aa ２８ The peptide product according to claim 1, 14, or 15, wherein is Asn, Gln, or U(X).

24. It has the following structure: His １ -aa ２ -Gln ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ -Asp ９ -Tyr １０ -Ser １１ -Lys １２ -Tyr １３ -Leu １４ -Asp １５ -aa １６ -aa １７ -Ala １８ -Ala １９ -Lys ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Gln ２４ -Trp ２５ -Leu ２６ -Leu ２７ -Asn ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.775)、 During the ceremony, aa ２ is Gly or Aib, aa １６ These are Glu, Ala, and Aib. aa １７ is Lys, or U(X), aa ２７ The peptide product according to claim 1, 14, or 15, wherein is Leu or Val.

25. It has the following structure: His １ -aa ２ -Gln ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ -Asp ９ -Tyr １０ -Ser １１ -Lys １２ -Tyr １３ -Leu １４ -Asp １５ -aa １６ -aa １７ -Arg １８ -Ala １９ -aa ２０ -Asp ２１ -Phe ２２ -aa ２３ -aa ２４ -Trp ２５ -Leu ２６ -aa ２７ -aa ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.776) During the ceremony, aa ２ is Gly or Aib, aa １６ is Glu, Ser, Ala, or Aib, aa １７ is Arg, hArg, or Gln, aa ２０ is Lys, or U(X), aa ２３ is Ile or Val, aa ２４ is Ala, Gln, or U(X), aa ２７ is Leu, or Val, and, aa ２８ The peptide product according to claim 1, 14, or 15, wherein is Asn, Gln, or U(X).

26. It has the following structure: _) １  ２ . ３ -ly ４ ﹁) ５ ________________ ６ ﹁) ７ ) ８ asp ９ )) １０ ) １１ yys １２ )) １３ eu １４ asp １５  １６  １７ ________________ １８ _____________ １９  ２０ asp ２１ ________________ ２２  ２３  ２４ ) ２５ eu ２６  ２７  ２８ ﹁) ２９ H ２ 4 (SEQ.ID.NO.777) During the ceremony, aa ２ is Gly or Aib, aa １６ is Glu, Ala, or Aib, aa １７ is Arg, hArg, or Gln, aa ２０ is Lys, or U(X), aa ２３ is Ile or Val, aa ２４ is GLN or ALA, aa ２７ is Leu or Val, aa ２８ The peptide product according to claim 1, 14, or 15, wherein is Asn or Glun.

27. aa １６ and aa ２０ The peptide product according to claim 1, 14, or 15, wherein the peptide is cyclized to form a lactam bond.

28. It has the following structure: _) １  ２ . ３ -ly ４ ﹁) ５ ________________ ６ ﹁) ７ ) ８ asp ９ )) １０ ) １１ yys １２ )) １３ eu １４ asp １５  １６  １７ _____________ １８ _____________ １９  ２０ lu ２１ ________________ ２２ le ２３ _________________________________ ２４ ) ２５ eu ２６  ２７  ２８ ﹁) ２９ H ２ 4 (SEQ.ID.NO.778) During the ceremony, aa ２ is Aib or Gly, aa １６ and aa ２０ Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond. aa １７ is Arg, hArg, or Gln, aa ２７ is Met, Val, Leu, or Nle, aa ２８ is Asn or Gln, X comprises a glucuronyl class portion prepared from 1-alkylbeta-D-glucoside, 1-alkylbeta-D-maltoside, 1-alkylbeta-D-melibioside, etc., and alkyl is C ８ -C ２０ The peptide product according to claim 23 or 27, wherein the linear alkyl chain is a peptide product according to claim 23 or 27.

29. It has the following structure: His １ -aa ２ -Gln ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ -Asp ９ -Tyr １０ -Ser １１ -Lys １２ -Tyr １３ -Leu １４ -Asp １５ -aa １６ -aa １７ -Ala １８ -Ala １９ -Lys ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Ala ２４ -Trp ２５ -Leu ２６ -Leu ２７ -Asn ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.779) During the ceremony, aa ２ is Aib or Gly, aa １６ is Glu, Ala, or Aib, and, aa １７ is Lys or Lys(N-omega-X), Alkyl is C ８ -C ２０ The peptide product according to claim 23 or 27, wherein the linear alkyl chain is a peptide product according to claim 23 or 27.

30. It has the following structure: _) １  ２ . ３ -ly ４ ﹁) ５ ________________ ６ ﹁) ７ ) ８ asp ９ )) １０ ) １１ yys １２ )) １３ eu １４ asp １５  １６  １７ ________________ １８ _____________ １９  ２０ asp ２１ ________________ ２２  ２３  ２４ ) ２５ eu ２６  ２７  ２８ ﹁) ２９ H ２ 4 (SEQ.ID.NO.780) During the ceremony, aa ２ is Gly or Aib, aa １６ is Glu, Ala, or Aib, aa １７ is Arg or hArg, aa ２０ is Lys or Lys(N-omega-X), aa ２３ is Ile or Val, aa ２４ is GLN or ALA, aa ２７ is Leu, or Val, and, aa ２８ is Asn or Gln, Alkyl is C ８ -C ２０ The peptide product according to claim 1, 14, 15, or 26, wherein the linear alkyl chain is...

31. It has the following structure: His １ -aa ２ -Gln ３ -Gly ４ -Thr ５ -aa ６ -Thr ７ -Ser ８ -Asp ９ -aa １０ -aa １１ -Z; (SEQ.ID.NO.781) During the ceremony, aa ２ It is Gly, Aib, or MePro, aa ６ is Phe, 2FPhe, MePhe, or 2FMePhe, aa １０ is Tyr, Nal2, Bip, or Bip2EtMeO, and, aa １１ is Lys or Lys(N-omega-X), Alkyl is C ８ -C ２０ The peptide product according to claim 1, 14, or 15, wherein the linear alkyl chain is...

32. The peptide product according to any one of claims 1 to 31, wherein X comprises an alkyl chain of dodecyl, tetradecyl, hexadecyl, or octadecyl.

33. A compound selected from the compounds in Table 1 of Figure 1, Table 2 of Figure 2, or Table 3 of Figure 3.

34. A pharmaceutical composition comprising a therapeutically effective amount of a peptide product or an acceptable salt thereof according to any one of claims 1 to 33, and at least one pharmaceutically acceptable carrier or excipient.

35. A method for treating a disease related to insulin resistance, comprising the step of administering a peptide product according to any one of claims 1 to 33 to an individual.

36. Amino acid residue aa of SEQ. ID. NO. 1 １ -aa ２７ A method for treating diabetes in an individual, comprising the step of administering to the individual a therapeutically effective amount of a glucagon analog containing the above.

37. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ２８ The method according to claim 35, including the method described in claim 35.

38. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ２９ The method according to claim 35, including the method described in claim 35.

39. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ３０ The method according to claim 35, including the method described in claim 35.

40. The glucagon analog is modified with the surfactant X of formula I, 【Transformation 8】 During the ceremony: R １ａ Independently, at each occurrence, there is a single bond, H, saccharide, substituted or unsubstituted C. １ -C ３０ The moiety contains an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid core. R １ｂ , R １ｃ , and R １ｄ Each of these elements, independently, represents a single bond, H, substituted or unsubstituted C at each occurrence. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, W １ These are independent, and at each occurrence, -CH ２ -ien-CH ２ -O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH ２ -S-, W ２ ha-O-, -CH ２ -, or -S-, R ２ Independently, at each occurrence, U is a single bond, H, substitution, or unsubstituted C. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, -NH, -S-, -triazolo-, -NH(C=O)-CH ２ -, - (CH ２ ) ｍ - It is maleimide, n is 1, 2, or 3, and, The method according to any one of claims 34-37, wherein m is 1-10.

41. The method according to any one of claims 34 to 38, wherein the administration of the glucagon analog causes weight loss.

42. Amino acid residue aa of SEQ. ID. NO. 1 １ -aa １７ A method for treating a cardiovascular disease in an individual, comprising the step of administering to the individual a therapeutically effective amount of a glucagon analog containing [a specific compound].

43. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ２８ The method according to claim 40, including the method described in claim 40.

44. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ２９ The method according to claim 40, including the method described in claim 40.

45. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ３０ The method according to claim 40, including the method described in claim 40.

46. The glucagon analog is modified with the surfactant X of formula I, 【Chemistry 9】 During the ceremony: R １ａ Independently, at each occurrence, there is a single bond, H, saccharide, substituted or unsubstituted C. １ -C ３０ The moiety contains an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid core. R １ｂ , R １ｃ , and R １ｄ Each of these elements, independently, represents a single bond, H, substituted or unsubstituted C at each occurrence. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, W １ These are independent, and at each occurrence, -CH ２ -ien-CH ２ -O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH ２ -S-, W ２ ha-O-, -CH ２ -, or -S-, R ２ Independently, at each occurrence, U is a single bond, H, substitution, or unsubstituted C. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, -NH, -S-, -triazolo-, -NH(C=O)-CH ２ -, - (CH ２ ) ｍ - It is maleimide, n is 1, 2, or 3, and, The method according to any one of claims 40-43, wherein m is 1-10.

47. The method according to any one of claims 40 to 44, wherein the cardiovascular disease is associated with an ischemic event.

48. Amino acid residue aa of SEQ. ID. NO. 1 １ -aa ２７ A method for treating diabetes in an individual, comprising the step of administering to the individual a therapeutically effective amount of a glucagon analog containing the above.

49. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ２８ The method according to claim 46, including the method described in claim 46.

50. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ２９ The method according to claim 46, including the method described in claim 46.

51. The aforementioned glucagon analog is the amino acid residue aa of SEQ. ID. NO.

1. １ -aa ３０ The method according to claim 46, including the method described in claim 46.

52. The glucagon analog is modified with the surfactant X of formula I, 【Chemistry 10】 During the ceremony: R １ａ Independently, at each occurrence, there is a single bond, H, saccharide, substituted or unsubstituted C. １ -C ３０ The moiety contains an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid core. R １ｂ , R １ｃ , and R １ｄ Each of these elements, independently, represents a single bond, H, substituted or unsubstituted C at each occurrence. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, W １ These are independent, and at each occurrence, -CH ２ -ien-CH ２ -O-, -(C=O), -(C=O)-O-, -(C=O)-NH-, -(C=S)-, -(C=S)-NH-, or -CH ２ -S-, W ２ ha-O-, -CH ２ -, or -S-, R ２ Independently, at each occurrence, U is a single bond, H, substitution, or unsubstituted C. １ -C ３０ Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, -NH, -S-, -triazolo-, -NH(C=O)-CH ２ -, - (CH ２ ) ｍ - It is maleimide, n is 1, 2, or 3, and, The method according to any one of claims 46-49, wherein m is 1-10.

53. The method according to any one of claims 46 to 50, wherein the administration of the glucagon analog causes weight loss.

54. It has the following structure: His １ -aa ２ -Gln ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ -Asp ９ -Tyr １０ -Ser １１ -Lys １２ -Tyr １３ -Leu １４ -Asp １５ -Glu １６ -U(X) １７ -Ala １８ -Ala １９ -Lys ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Gln ２４ -Trp ２５ -Leu ２６ -Leu ２７ -aa ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.795) During the ceremony, aa ２ is Gly or Aib, aa ２８ The peptide product according to claim 1, 14, 15, or 23, wherein is Asn or Gln.

55. It has the following structure: _) １ ) ２ . ３ -ly ４ ﹁) ５ ________________ ６ ﹁) ７ ) ８ asp ９ )) １０ ) １１ yys １２ )) １３ eu １４ asp １５  １６ . １７ 1. １８ _____________ １９  ２０ lu ２１ ________________ ２２ le ２３ _________________________________ ２４ ) ２５ eu ２６ eu ２７  ２８ ﹁) ２９ H ２ 4 (SEQ.ID.NO.796) During the ceremony, aa １６ and aa ２０ Each is individually Lys or Glu, and is cyclized by its side chain to form a lactam bond, and, aa ２８ is Asn or Gln, X comprises a glucuronyl class portion prepared from 1-alkylbeta-D-glucoside, 1-alkylbeta-D-maltoside, 1-alkylbeta-D-melibioside, or the corresponding alpha-glycoside, where alkyl is C ８ -C ２０ The peptide product according to any one of claims 23, 27, or 28, wherein the linear alkyl chain is...

56. It has the following structure: _) １ ) ２ . ３ -ly ４ ﹁) ５ ________________ ６ ﹁) ７ ) ８ asp ９ )) １０ ) １１ yys １２ )) １３ eu １４ asp １５ lu ＊ １６ . １７ _____________ １８ _____________ １９ yys ＊ ２０ lu ２１ ________________ ２２ le ２３ _________________________________ ２４ ) ２５ eu ２６ eu ２７ . ２８ ﹁) ２９ H ２ 4 (SEQ.ID.NO.797) During the ceremony, aa １６ and aa ２０ It is cyclized by its side chain to form a lactam bond, and, X comprises a glucuronyl class portion prepared from 1-alkylbeta-D-glucoside, 1-alkylbeta-D-maltoside, 1-alkylbeta-D-melibioside, or the corresponding alpha-glycoside, where alkyl is C ８ -C ２０ The peptide product according to any one of claims 23, 27, 28, or 55, wherein the linear alkyl chain is...

57. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ - Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-dodecylbeta-D-glucuronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.601).

58. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ - Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-tetradecylbeta-D-glucuronil)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.602).

59. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ - Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-hexadecylbeta-D-glucuronil)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.603)

60. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ -Gln ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ -Asp ９ -Tyr １０ -Ser １１ -Lys １２ -Tyr １３ -Leu １４ -Asp １５ -Glu ＊ １６ -Gln １７ Ala １８ -Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-octadecyl beta-D-glucuronyl)) ２４ -Trp ２５ -Leu ２６ -Leu ２７ -Gln ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.604).

61. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-octylbeta-D-meribiouronyl) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ. ID. NO. 630).

62. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ -Gln ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ -Asp ９ -Tyr １０ -Ser １１ -Lys １２ -Tyr １３ -Leu １４ -Asp １５ -Glu ＊ １６ -Gln １７ Ala １８ -Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-dodecyl-beta-D-melibiouronyl)) ２４ -Trp ２５ -Leu ２６ -Leu ２７ -Gln ２８ -Thr ２９ -NH ２ ; (SEQ. ID. NO. 631).

63. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-tetradecylbeta-D-meribiouronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ. ID. NO. 632).

64. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-hexadecylbeta-D-meribiouronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ (SEQ.ID.NO.633).

65. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-octadecylbeta-D-meribiouronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ. ID. NO. 634).

66. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-hexadecyl alpha-D-meribiouronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.805).

67. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-tetradecyl alpha-D-meribiouronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ - GLN ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.819).

68. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-hexadecyl alpha-D-meribiouronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ - GLN ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.820).

69. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ Lys(N-Omega(1-Octadecyl Alpha-D-Melibiouronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ - GLN ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.821).

70. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-dodecyl alpha-D-meribiouronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-dodecylbeta-D-glucouronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1099).

71. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-tetradecyl alpha-D-meribiouronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-tetradecylbeta-D-glucouronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1100).

72. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-hexadecyl alpha-D-meribiouronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-hexadecylbeta-D-glucouronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1101).

73. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-(13-carboxyl-tridecyloxy)beta-D-glucuronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ - GLN ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1102).

74. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-(15-carboxyl-pentadecyloxy)beta-D-glucuronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ - GLN ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1103).

75. A peptide product according to any one of claims 23, 27, 28, 54, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ -Lys(N-omega(1-(17-carboxyl-heptadecyloxy)beta-D-glucuronyl)) １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ - GLN ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1104).

76. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-(13-carboxyl-tridecyloxy)beta-D-glucuronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1105).

77. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ - Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-(15-carboxyl-pentadecyloxy)beta-D-glucuronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1106).

78. A peptide product according to any one of claims 23, 27, 28, 55, or 56, having the following structure: His １ -Aib ２ - GLN ３ -Gly ４ -Thr ５ -Phe ６ -Thr ７ -Ser ８ - Asp ９ - Tyr １０ -Ser １１ -Lys １２ - Tyr １３ - Leu １４ - Asp １５ -Glu ＊ １６ - GLN １７ - Ala １８ - Ala １９ -Lys ＊ ２０ -Glu ２１ -Phe ２２ -Ile ２３ -Lys(N-omega(1-(17-carboxyl-heptadecyloxy)beta-D-glucuronyl)) ２４ -Trp ２５ - Leu ２６ - Leu ２７ - GLN ２８ -Thr ２９ -NH ２ ; (SEQ.ID.NO.1107).

79. It has the structure of formula III-A, aa １ -aa ２ -aa ３ -aa ４ -aa ５ -aa ６ -aa ７ -aa ８ -aa ９ -aa １０ -aa １１ -aa １２ -aa １３ -aa １４ -aa １５ -aa １６ -aa １７ -aa １８ -aa １９ -aa ２０ - aa ２１ -aa ２２ -aa ２３ -aa ２４ -aa ２５ -aa ２６ -aa ２７ -aa ２８ -aa ２９ -Z Formula III-A (SEQ. ID. NO. 2) During the ceremony: Z is OH or -NH-R ３ And, R ３ is H, or substituted or unsubstituted C １ -C １２ Alkyl or PEG chains less than 10 Da, aa １ His is His, aa ２ It is Aib, aa ３ is Gln, aa ４ It is Gly, aa ５ is Thr, aa ６ is Phe, aa ７ is Thr, aa ８ is Ser, aa ９ It is Asp, aa １０ is Tyr, Glu, Lys, or U(X), aa １１ is Ser, aa １２ is Lys or Glu, aa １３ It is Tyr, aa １４ is Leu, Glu, or Lys, aa １５ It is Asp, aa １６ is Glu or Lys, aa １７ is Glun, Glu, or U(X), aa １８ It is Ala, aa １９ It is Ala, aa ２０ is Lys, Glu, or U(X), aa ２１ is Glu, aa ２２ It is Phe, aa ２３ is Ile, aa ２４ is Glun, Glu, or U(X), aa ２５ is Trp, aa ２６ is Leu, aa ２７ is Leu, aa ２８ is Glu or Glun, aa ２９ is Thr, Here, aa １６ and aa ２０ , or aa １０ and aa １４ , or aa １２ and aa １６ It can be voluntarily cyclized by its side chain to form a lactam bond. However, aa １０ aa １７ aa ２０ , or aa ２４ The peptide product according to claim 1 or 3, wherein one or at least one of is a natural or unnatural amino acid U covalently bonded to X.

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