Improved peptide formulations

Abstract

To provide peptides and / or proteins that allow for improved stability, bioavailability, selectivity and duration of the effect in peptide and / or protein-based therapies.SOLUTION: Provided is a peptide product comprising a surfactant X covalently linked to a peptide, where the peptide comprises a linker amino acid U and at least one other amino acid, X comprises A and B as shown in the formula where A is a hydrophobic group; and B is a hydrophilic group covalently linked to the peptide via a linker amino acid U.SELECTED DRAWING: None

Description

[Technical Field]

[0001] <Cross reference> This application claims priority to U.S. Provisional Patent Application No. 61 / 487,636, filed May 18, 2011, U.S. Provisional Patent Application No. 61 / 487,638, filed May 18, 2011, U.S. Provisional Patent Application No. 61 / 543,725, filed October 5, 2011, and U.S. Provisional Patent Application No. 61 / 543,721, filed October 5, 2011, which are incorporated herein by reference in their entireties.

[0002] Covalently modified peptide and protein analogs allow for improved pharmaceutical properties of peptide and / or protein-based therapeutics. Summary of the Invention [Problem to be solved by the invention]

[0003] Formulations of peptides and / or proteins suffer from several limitations in their use in medicine, namely, a short duration of action, poor bioavailability, and a lack of receptor subtype selectivity (Nestor, JJ, Jr. (2007) Comprehensive Medicinal Chemistry II 2: 573-601). Furthermore, peptides and / or proteins are unstable in formulations and subject to aggregation. In some instances, aggregation of peptides and / or proteins leads to the development of immune responses against native or heterologous peptides or proteins. [Means for solving the problem]

[0004] Described herein are methods and reagents for covalently modifying peptides and / or proteins to produce products with improved pharmaceutical properties. In some instances, the covalently modified peptides and / or proteins enable improved stability, bioavailability, selectivity, and duration of effect in peptide- and / or protein-based therapeutics.

[0005] Described herein are certain covalently modified peptides and / or proteins with improved pharmaceutical properties. In some instances, these covalently modified peptides and / or proteins enable improved stability, bioavailability, selectivity, and duration of effect in peptide- and / or protein-based therapeutics.

[0006] In some embodiments, the covalently modified peptides and / or proteins described herein are attached to glycoside surfactants. In one aspect, the covalently modified peptides and / or proteins are attached to alkyl glycosides. In one aspect, the covalently modified peptides and / or proteins are attached to alkyl glycoside surfactants, where the peptide and / or protein is attached to a glycoside in the surfactant, which is then attached to a hydrophobic group and / or alkyl group. In some embodiments, provided herein are reagents and intermediates for the covalent modification of peptides and / or proteins via the incorporation of surfactants such as alkyl glycosides.

[0007] Provided herein are peptide products comprising a surfactant X covalently attached to a peptide, the peptide comprising a linker amino acid U and at least one other amino acid:

[0008] [ka]

[0009] wherein X is:

[0010] [ka]

[0011] and During the ceremony, A is a hydrophobic group, and B is a hydrophilic group covalently attached to the peptide via the linker amino acid U.

[0012] In some embodiments, the peptide product is synthesized by reacting a functionalized surfactant with a peptide as described herein. In some embodiments, the peptide product is synthesized by reacting a functionalized surfactant with a reversibly protected linker amino acid as described herein, followed by reaction with one or more amino acids to form the surfactant-modified peptide product. In some embodiments, U is the terminal amino acid of the peptide. In some embodiments, U is a non-terminal amino acid of the peptide.

[0013] In some embodiments, A is a substituted or unsubstituted C-C 30 In some embodiments, A is a substituted or unsubstituted C-C alkyl chain, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a moiety containing a steroid nucleus. 20 In some embodiments, A is a substituted or unsubstituted C 10 -C 20 It is an alkyl chain, a substituted or unsubstituted 1-alkoxyaryl group, a substituted or unsubstituted 1-aralkyl group, or a moiety containing a steroid nucleus.

[0014] In some embodiments, B is a polyol. In some embodiments, the polyol is a saccharide. In some embodiments, the saccharide is a monosaccharide, a disaccharide, or a polysaccharide. In some embodiments, the saccharide is selected from glucose, mannose, maltose, glucuronic acid, galacturonic acid, diglucuronic acid, and maltouronic acid.

[0015] In some embodiments, the surfactant is a 1-alkyl glycoside class surfactant.

[0016] In some embodiments, the hydrophilic group in the surfactant is attached to the peptide via an amide bond.

[0017] In some embodiments of the peptide product, the surfactant 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-isopropyl ... 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-hexadecyl beta-D-diglucuronic acid, 1-tetradecyl beta-D-diglucuronic acid, 1-dodecyl beta-D-diglucuronic acid, 1-decyl beta-D-diglucuronic acid, 1-octyl beta-D-diglucuronic acid, or functionalized 1-eicosyl (ecosyl) beta-D-glucose, 1-octadecyl beta-D-glucose, 1-hexadecyl beta-D-glucuronic acid beta-D-maltoside, 1-tetradecyl beta-D-glucose, 1-dodecyl beta-D-glucose, 1-decyl beta-D-glucose, 1-octyl beta-D-glucose, 1-eicosyl beta-D-maltoside, 1-octadecyl beta-D-maltoside, 1-hexadecyl beta-D-maltoside, 1-dodecyl beta-D-maltoside, 1-decyl beta-D-maltoside, 1-octyl beta-D-maltoside, etc., and peptide products are prepared by the formation of a bond between the aforementioned groups and a group on the peptide (e.g., a -COOH group on the aforementioned group and an amino group on the peptide).

[0018] In some embodiments, the combination of a hydrophilic group with a hydrophobic group produces a surfactant. In some embodiments, the surfactant is an ionic surfactant. In some embodiments, the surfactant is a non-ionic surfactant. In some embodiments, the hydrophobic group is a substituted or unsubstituted C-C 30 It is an alkyl chain or an aralkyl chain. In some embodiments, the hydrophobic group is a chain with mixed hydrophobic and hydrophilic properties, such as a polyethylene glycol (PEG) group.

[0019] In some embodiments, the linker amino acid is a naturally occurring D-amino acid or an L-amino acid. In some embodiments, the linker amino acid is an unnatural amino acid. In some embodiments, the linker amino acid is selected from Lys, Cys, Orn, Asp, Glu, or an unnatural amino acid that comprises a functional group that is used to covalently attach to surfactant X. In some embodiments, the linker amino acid is selected from Lys, Cys, Orn, or an unnatural amino acid that comprises a functional group that is used to covalently attach to surfactant X. In some embodiments, the functional group that is used to covalently attach to surfactant X is -NH, -SH, -OH, -N, haloacetyl, a-(CH) m -maleimide or acetylene group, and m is 1-10.

[0020] In some embodiments, the peptide is an opioid peptide. In some embodiments, the peptide and / or protein product contains a covalently linked alkyl glycoside. In some such embodiments, the peptide and / or protein product contains a covalently linked alkyl glycoside that is a 1-O-alkyl glucuronic acid of the alpha or beta configuration. In some such embodiments, the peptide and / or protein product contains a covalently linked alkyl glycoside that is a 1-O-alkyl glucuronic acid, the alkyl chain being C1 to C6. 20 is an alkyl chain.

[0021] In some embodiments, the peptide product has the structure of formula IA:

[0022] [ka]

[0023] During the ceremony: Each of aa1, aa2, aa3, aa4, and aa5 is independently selected from the group consisting of: a non-existent, a D or L natural or unnatural amino acid, an N-alkylated amino acid, an N-acetylated amino acid, a CαR 3 an amino acid, a Ψ amino acid, or a linker amino acid U covalently attached to a surfactant X; Z is -OH, -NH2 or -NHR 3 and; Each R 3 are independently substituted or unsubstituted C1-C 12 a branched or linear alkyl, a PEG chain of less than 10 Da, or a substituted or unsubstituted aralkyl chain; with the proviso that one or at least one of aa1, aa2, aa3, aa4, and aa5 is a linker amino acid U covalently attached to surfactant X; and further provided that aa1, aa2, aa3, aa4, and aa5 are not all absent.

[0024] In one embodiment, provided herein is a peptide product having the structure of Formula II:

[0025] [ka]

[0026] During the ceremony: Each of aa1, aa2, aa3, aa4, and aa5 is independently selected from the group consisting of: a non-existent, a D or L natural or unnatural amino acid, an N-alkylated amino acid, an N-acetylated amino acid, a CαR 3 an amino acid, a Ψ amino acid, or a linker amino acid U covalently attached to a surfactant X; Z is -OH, -NH2 or -NHR 3 and; Each R 3 are independently substituted or unsubstituted C1-C 12 a branched or linear alkyl, a PEG chain of less than 10 Da, or a substituted or unsubstituted aralkyl chain; and X is

[0027] [ka]

[0028] and During the ceremony, A is a hydrophobic group; and B is a hydrophilic group covalently attached to the peptide via the linker amino acid U; with the proviso that one or at least one of aa1, aa2, aa3, aa4, and aa5 is a linker amino acid U covalently attached to surfactant X; and further provided that aa1, aa2, aa3, aa4, and aa5 are not all absent.

[0029] In one aspect, provided herein is a peptide product having the structure of Formula III:

[0030] [ka]

[0031] During the ceremony: aa1 is Tyr, Dmt, NR 3 -Tyr, NR 3 -Dmt, N-(R 3 )2-Tyr, or N-(R 3 )2-Dmt; aa2 is Pro, D-Arg, DU(X), D-Ala, Tic, or Tic(Ψ[CH2-NH]); aa3 is Phe, Trp, Tmp, D- or L-Nal(1), D- or L-Nal(2), CαMePhe, or Ψ-Phe; aa4 is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), U(X), D- or L-CαMeU(X); aa5 is absent or is Pro, Aib, U(X), D- or L-CαMeU(X); and U is a dibasic natural or unnatural amino acid, a thiol-containing natural or unnatural amino acid, an unnatural amino acid containing an -N group, an acetylene-containing unnatural amino acid, or an -NH-C(=O)-CH-Br or -(CH) group, used to covalently attach to surfactant X. m -maleimide-containing unnatural amino acids, where m is 1-10.

[0032] In some embodiments of the peptide products of Formula I, Formula II, or Formula III described above and herein, X has the structure:

[0033] [ka]

[0034] During the ceremony: A is a substituted or unsubstituted C1-C 30 a moiety containing an alkyl chain, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid nucleus; R 1b , R 1c , and R 1d is independently at each occurrence H, a protecting group, or a substituted or unsubstituted C-C 30 alkyl groups; 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-, or -S-; R 2 is a single bond, a C2-C4 alkene, a C2-C4 alkyne, or -(CH2) m -maleimide; and m is 1-10.

[0035] In some embodiments of the peptide products of Formula I, Formula II, or Formula III described above and herein, X has the following structure:

[0036] [ka]

[0037] Thus, in the embodiments described above, R 2 is a single bond.

[0038] In some embodiments of the peptide products of Formula I, Formula II, or Formula III described above and herein, X has the following structure:

[0039] [ka]

[0040] In some embodiments of the peptide products of Formula I, Formula II, or Formula III described above and herein, X has the structure:

[0041] [ka]

[0042] During the ceremony: A is a substituted or unsubstituted C1-C 30 an alkyl group or a moiety containing a steroid nucleus; R 1b , R 1c and R 1d is independently at each occurrence H, a protecting group, or a substituted or unsubstituted C-C 30 is an alkyl group; W 1 is —(C═O)—NH—; W 2 is -O-; R 2 is a single bond.

[0043] In some embodiments of the peptide products of Formula I, Formula II, or Formula III described above and herein, X has the structure:

[0044] [ka] During the ceremony: A is a substituted or unsubstituted C1-C 30 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 is a single bond.

[0045] In some embodiments of the peptide products of Formula I, Formula II, or Formula III described above and herein, X is as described above and A is a substituted or unsubstituted C-C 20 It is an alkyl group.

[0046] Also contemplated herein is an alternative embodiment wherein X in Formula I has the following structure:

[0047] [ka]

[0048] For example, in exemplary embodiments of the structure of X described above, W 1 is -S- and R 2 is C1-C 30 is an alkyl group, and W 2 is S and R 1a is W 2 and the appropriate portion of an amino acid residue U in the peptide (for example, a thiol group in a cysteine ​​residue of the peptide forms a thioether with X).

[0049] In another exemplary alternating embodiment of the structure of X described above, W 1 is -S- and R 2 is C1-C 30 is an alkyl group, and W 2 is O and R 1a is W 2 and the appropriate portion of the amino acid residue U in the peptide (for example, the hydroxyl group in a serine or threonine residue of the peptide forms an ether with X).

[0050] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 149): During the ceremony: U is a dibasic natural or unnatural amino acid; X is a surfactant of the 1-alkyl glycoside class, where 1-alkyl is a substituted or unsubstituted C1-C 20 alkyl or substituted or unsubstituted alkoxyaryl substituents; Z is NH; aa1 is Tyr, Dmt, Nα-Me-Tyr, Nα-Me-Tyr, N,Nα-diMe-Tyr, or N,Nα-diMe-Dmt; aa2 is Pro, D-Arg, DU(X), D-Ala, Tic, or Tic(Ψ[CH2-NH]); aa3 is Phe, Trp, Tmp, D- or L-Nal(1), D- or L-Nal(2), CαMePhe, or Ψ-Phe; aa4 is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), U(X), D- or L-CαMeU(X); aa5 is absent, Pro, Aib, U(X), D- or L-CαMeU(X).

[0051] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 150): During the ceremony: X consists of 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid; Z is NH; aa1 is Dmt; aa2 is Pro, D-Lys(X), Tic, or Tic(Ψ[CH2-NH]); aa3 is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), or Ψ-Phe; aa4 is Phe, D- or L-Nal(1), D- or L-Nal(2), or Lys(X); aa5 is absent, Pro, Aib, Lys(X), D- or L-CαMeLys(X).

[0052] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO.151): During the ceremony: aa1 is Dmt; aa2 is Pro; aa3 is Phe, or Tmp; aa4 is Phe, or Lys(X); aa5 is absent, Aib, Lys(X), D- or L-CαMeLys(X).

[0053] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 152): During the ceremony: aa1 is Dmt; aa2 is Pro; aa3 is Phe, or Tmp; aa4 is Phe, or Lys(X); aa5 is absent or Lys(X).

[0054] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 153): During the ceremony: U is a dibasic natural or unnatural amino acid; X is a surfactant of the 1-alkylglycoside class, where the 1-alkyl group of the 1-alkylglycoside is a substituted or unsubstituted C1-C 20 alkyl or substituted or unsubstituted alkoxyaryl substituents; aa1 is Tyr, Dmt, NR 3 -Tyr, NR 3 -Tyr, N-(R 3 )2-Tyr, or N-(R 3 )2-Dmt; aa2 is D-Arg, or DU(X); aa3 is Phe, D- or L-Nal(1), D- or L-Nal(2), or Tmp; aa4 is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), U(X), D- or L-CαMeU(X); aa5 is absent, Pro, or U(X).

[0055] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 154): During the ceremony: X is a surfactant of the 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid class; aa1 is Dmt, Nα-Me-Dmt, or N,Nα-Me-Dmt); aa2 is D-Arg, D-Lys(X), or D-Orn(X); aa3 is Phe, or Tmp; aa4 is Phe, Tmp, Lys(X), or Orn(X); aa5 is absent, Pro, Lys(X), or Orn(X).

[0056] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 155): During the ceremony: U is a dibasic natural or unnatural amino acid; X is a surfactant of the 1-alkyl glycoside class, where 1-alkyl is a substituted or unsubstituted C1-C 20 alkyl or substituted or unsubstituted alkoxyaryloxy substituents; Z is NH; aa1 is Tyr, Dmt, NR 3 -Tyr, NR 3 -Dmt, N-(R 3 )2-Tyr, or N-(R 3 )2-Dmt; aa2 is Tic, or Tic(Ψ[CH2-NH]); aa3 is Ψ-Phe when aa2 is Tic(Ψ[CH2-NH]); aa4 is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), or U(X); aa5 is absent, Pro, Aib, or U(X).

[0057] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 156): During the ceremony: X is a surfactant of the 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid class; aa1 is Tyr, Dmt, Nα-Me-Tyr, Nα-alkyl-Dmt, N,Nα-diMe-Tyr, or N,Nα-Me-Dmt; aa2 is Tic or Tic(Ψ[CH2-NH]); aa3 is Ψ-Phe when aa2 is Tic(Ψ[CH2-NH]), Phe, or TMP; aa4 is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), Lys(X), or Orn(X); aa5 is absent, Aib, Lys(X), or Orn(X).

[0058] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 157): During the ceremony: X consists of 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid; aa2 is Tic, or Tic(Ψ[CH2-NH]); aa3 is Phe or Ψ-Phe; aa4 is Lys(X); aa5 does not exist.

[0059] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 158): During the ceremony: X consists of 1-alkyl glucuronic acid; aa2 is Tic: aa3 is Phe; aa4 is Lys(X); aa5 does not exist.

[0060] In some embodiments, the peptide product of Formula III is the product (SEQ.ID.NO. 159): During the ceremony: X consists of a 1-alkyl glucuronic acid selected from 1-methyl beta-D-glucuronic acid, 1-octyl beta-D-glucuronic acid, 1-dodecyl beta-D-glucuronic acid, 1-tetradecyl beta-D-glucuronic acid, 1-hexadecyl beta-D-glucuronic acid, and 1-octadecyl beta-D-glucuronic acid; aa2 is Tic; aa3 is Phe; aa4 is Lys(X); aa5 does not exist.

[0061] In some embodiments, the peptide product of formula III is H-Dmt-Tic-Phe-Lys(N-epsilon-1-methylbeta-D-glucuronyl)-NH2 (SEQ.ID.NO.78).

[0062] In some embodiments, the peptide product of formula III is H-Dmt-Tic-Phe-Lys(N epsilon-1-octyl beta-D-glucuronyl)-NH2 (SEQ. ID. NO. 80).

[0063] In some embodiments, the peptide product of formula III is H-Dmt-Tic-Phe-Lys(N epsilon-1-dodecyl beta-D-glucuronyl)-NH2 (SEQ. ID. NO. 79).

[0064] In some embodiments, the peptide product of formula III is H-Dmt-Tic-Phe-Lys(N epsilon-1-tetradecyl beta-D-glucuronyl)-NH2 (SEQ. ID. NO. 160).

[0065] In some embodiments, the peptide product of formula III is H-Dmt-Tic-Phe-Lys(N epsilon-1-hexadecyl beta-D-glucuronyl)-NH2 (SEQ. ID. NO. 82).

[0066] In some embodiments, the peptide product of formula III is H-Dmt-Tic-Phe-Lys(N-epsilon-1-octadecylbeta-D-glucuronyl)-NH2 (SEQ.ID.NO.83).

[0067] In some embodiments, the peptide product is biologically active.

[0068] In specific embodiments, provided herein is a compound selected from the compounds of Table 1 in Figures 1-1 through 1-7.

[0069] Also provided herein are pharmaceutical compositions comprising a therapeutically effective amount of a peptide product of Formula I, II or III, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0070] Provided herein are methods for treating pain comprising administering a therapeutically effective amount of a peptide product of Formula I, II or III or a compound of Table 1 of Figures 1-1 through 1-7.

[0071] A method for improving the pharmaceutical and therapeutic behavior of a peptide, thereby improving its duration of action, bioavailability, or its stability in a formulation, the method comprising covalently attaching a surfactant X to the peptide, wherein X is as described herein.

[0072] In some embodiments, the peptide products comprising covalently linked alkyl glycosides described herein are analogs of Leu-enkephalin. In some such embodiments, the peptide products contain covalently linked 1-O-alkyl β-D-glucuronic acid, and the peptides are analogs of Leu-enkephalin.

[0073] In some embodiments, the peptide product containing a covalently linked alkyl glycoside described herein is an analog of the opioid peptide DPDPE (Akiyama K, et al. (1985) Proc Natl Acad Sci USA 82:2543-7). In some such embodiments, the peptide product contains a covalently linked 1-O-alkyl glucuronic acid, and the peptide is an analog of the opioid peptide DPDPE.

[0074] In some embodiments, the peptide products containing covalently linked alkyl glycosides described herein are analogs of D-amino acids containing the naturally occurring peptides dermorphin (Melchiorri, P. and Negri, L. (1996) Gen Pharmacol 27: 1099-1107) or deltorphin (Erspamer, V., et al. (1989) Proc Natl Acad Sci USA 86:5188-92). In some such embodiments, the peptide product contains a covalently linked 1-O-alkyl β-D-glucuronic acid and the peptide is an analog of dermorphin or deltorphin.

[0075] In some embodiments, the peptide product containing a covalently linked alkyl glycoside is an analog of endomorphin-1 or -2. In some such embodiments, the peptide product contains a covalently linked 1-O-alkyl β-D-glucuronic acid, and the peptide is an analog of endomorphin (Lazarus, LH and Okada, Y. (2012) Expert Opin Ther Patents 22: 1-14).

[0076] In some embodiments, the peptide products containing covalently linked alkyl glycosides described herein are analogs of the opioid peptide dynorphin (James, I. F., et al. (1982) Life Sci 31:1331-4). In some such embodiments, the peptide product contains a covalently linked 1-O-alkyl β-D glucuronic acid, and the peptide is an analog of dynorphin.

[0077] In some embodiments, the side chain functional groups of two different amino acid residues are linked to form a cyclic lactam. For example, in some embodiments, a Lys side chain forms a cyclic lactam with the side chain of Glu. In some embodiments, such a lactam structure is reversed and formed from Glu and Lys. Such lactam bonds in some instances are known to stabilize alpha-helical structures in peptides (Condon, SM, et al. (2002) Bioorg Med Chem 10: 731-736).

[0078] In some embodiments, the side chain functional groups of two different amino acid residues having -SH-containing side chains are linked to form a cyclic disulfide. For example, in some embodiments, two penicillamine or two Cys side chains can be linked to constrain the conformation of the peptide to produce a longer duration of action or greater receptor selectivity (Akiyama K, et al. (1985) Proc Natl Acad Sci USA 82:2543-7).

[0079] In specific embodiments, the peptide product of Formula I, Formula II, or Formula III described above and herein has the following structure:

[0080] [ka]

[0081] In the formula, A is C1-C as described in Table 1 of Figures 1-1 to 1-7. 20 R' is a peptide as described in Table 1 of Figures 1-1 to 1-7, W of formula V 2 is -O-, and W of formula V 1 is —(C═O)NH— and is part of an amide bond to the peptide R′. In some such embodiments, A is C6-C 20In some such embodiments, A is a C-C alkyl chain. 10 In some such embodiments, A is an alkyl chain. 12 -C 20 In some such embodiments, A is an alkyl chain. 12 -C 18 It is an alkyl chain.

[0082] In the embodiments described above, the amino acid and / or amino moiety of peptide R′ (e.g., the amino group of an amino acid residue such as lysine, or a lysine within peptide R′) is used to form a covalent bond with a compound of the following structure:

[0083] [ka]

[0084] where A is C1-C2 as described above and in Table 1 of Figures 1-1 through 1-7. 20 It is an alkyl chain.

[0085] In such cases, the amino acid residue having an amino moiety (e.g., lysine in peptide R') used to form a covalent bond to the compound of Formula A described above is the linker amino acid U attached to surfactant X. Thus, as an example, Lys(C12) in Table 1 of Figures 1-1 through 1-7 has the following structure:

[0086] [ka]

[0087] Also contemplated within the scope of the embodiments presented herein are peptide products of Formula I resulting from malturonic acid-based surfactants prepared by covalent attachment of a peptide to either or both of the carboxylic acid groups. Thus, as one example, the peptide in Table 1 of Figures 1-1 through 1-7 includes a lysine linker amino acid attached to a malturonic acid-based surfactant, X, and having the following structure:

[0088] [ka]

[0089] It will be understood that in one embodiment, compounds of Formula I are prepared by attaching lysine to group X, followed by attaching additional amino acid residues and / or peptides to the lysine-X compound to obtain compounds of Formula I. It will be understood that other natural or unnatural amino acids described herein are also suitable for attachment to surfactant X, and for attaching additional amino acids / peptides to obtain compounds of Formula I. It will be understood that in another embodiment, compounds of Formula I are prepared by attaching a full-length or partial-length peptide to group X, followed by attaching additional amino acid residues and / or peptides to obtain compounds of Formula I.

[0090] Also provided herein is a pharmaceutical composition comprising a therapeutically effective amount of the peptide product described above, or an acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient. In some embodiments, the carrier is an aqueous-based carrier. In some embodiments, the carrier is a non-aqueous-based carrier. In some embodiments, the non-aqueous-based carrier is a hydrofluoroalkane-like solvent containing submicron anhydrous α-lactose, or other excipient.

[0091] Within the embodiments presented herein, reaction of amino acids and / or peptides comprising a nucleophile-bearing linker amino acid U with a group X comprising a leaving group or a functional group that can be activated to contain a leaving group, e.g., a carboxylic acid, or any other reactive group, is contemplated, thereby allowing covalent attachment of the amino acid and / or peptide to surfactant X via the linker amino acid U to provide a peptide product of Formula I.

[0092] Also contemplated within the embodiments presented herein is the reaction of amino acids and / or peptides containing a linker amino acid U bearing a leaving group or a functional group that can be activated to contain a leaving group, e.g., a carboxylic acid, or any other reactive group, with a group X containing a nucleophilic group, thereby allowing covalent attachment of the amino acid and / or peptide to surfactant X via the linker amino acid U to provide a peptide product of Formula I.

[0093] It will be appreciated that in one embodiment, compounds of formula I are prepared by reaction of a linker amino acid U with X, followed by attachment of additional residues to U to obtain the peptide product of formula I.

[0094] It will be appreciated that in an alternative embodiment, compounds of formula I are prepared by reaction of a suitable peptide containing a linker amino acid U with X, followed by optional attachment of additional residues to U to give the peptide product of formula I.

[0095] Further provided herein are certain intermediates and / or reagents suitable for synthesizing the peptide products described herein. In certain embodiments, such intermediates and / or reagents are functionalized surfactants that allow for covalent attachment to peptides. In certain embodiments, such intermediates are functionalized 1-alkylglycoside surfactants that allow for covalent attachment to peptides. In certain embodiments, such intermediates are functionalized 1-alkylglycoside surfactants linked to reversibly protected linker amino acids that allow for covalent attachment to other amino acids to form peptides. It will be understood that an appropriately functionalized surfactant is covalently linked to a peptide via reaction with an appropriately matched functional group on the linker amino acid.

[0096] In some embodiments, a suitable intermediate for the synthesis of the peptide products described herein is a compound of formula IV. Accordingly, provided herein are intermediates and / or compounds of formula IV:

[0097] [ka]

[0098] During the ceremony: R 1a is independently at each occurrence a single bond, H, a protecting group, a natural or unnatural amino acid, a substituted or unsubstituted C-C 30 an alkyl hydrophobic group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a moiety containing a steroid nucleus; R 1b , R 1c , and R 1d is independently at each occurrence a single bond, H, a protecting group, a natural or unnatural amino acid, a substituted or unsubstituted C-C 30 an alkyl hydrophobic group, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group; W 1is -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 H, a protecting group, a natural or unnatural amino acid, a substituted or unsubstituted C1-C 30 Alkyl hydrophobic groups, substituted or unsubstituted alkoxyaryl groups, substituted or unsubstituted aralkyl groups, -NH2, -SH, C2-C4-alkene, C2-C4-alkyne, -NH(C=O)-CH2-Br, -(CH2) m -maleimide, or -N3; n is 1, 2 or 3; m is 1-10.

[0099] In some embodiments, each natural or unnatural amino acid is independently a reversibly protected or free linker amino acid, hi some such embodiments, the linker amino acid is a reversibly protected or free lysine.

[0100] In some embodiments of Formula IV, n is 1; W 1 is -(C=O)-; R 1a is a substituted or unsubstituted C1-C 30 an alkyl hydrophobic group, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group; R 2 is a reversibly protected lysine in either the D- or L-configuration.

[0101] In some embodiments of Formula IV, n is 1; W 1 is -(C=O)-; R 1a is a substituted or unsubstituted C8-C 30an alkyl hydrophobic group, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group; R 2 is a reversibly protected lysine in either the D- or L-configuration.

[0102] In some such embodiments, R 1a is an octyl, decyl, dodecyl, tetradecyl, or hexadecyl group.

[0103] In some embodiments of Formula IV, n is 1; W 1 is —(C═O)—NH— or —(C═O)—O—; R 2 is a substituted or unsubstituted C1-C 30 an alkyl hydrophobic group, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group; R 1a is a reversibly protected serine or threonine in either the D- or L-configuration.

[0104] In some such embodiments, R 2 is an octyl, decyl, dodecyl, tetradecyl, or hexadecyl group.

[0105] In some embodiments of Formula IV, n is 1; m is 1-6; W 1 is -CH2-; R 1a is C1-C 30 an alkyl hydrophobic group, a 1-alkoxyaryl group, or a 1-aralkyl group; R 2 are -N3, NH2, -C2-alkyne, -(CH2) m -maleimide, NH—(C═O)—CH₂—Br, or NH—(C═O)—CH₂—I.

[0106] In some embodiments of Formula IV, n is 1; W 1 is -(C=O)-O-; R 2 is H, R 1a is a substituted or unsubstituted C1-C 30 It is an alkyl hydrophobic group.

[0107] In some embodiments of Formula IV, R 2 is attached to the peptide or is an amino acid residue in the peptide. In some such embodiments, R 2 is a reversibly protected lysine.

[0108] In some embodiments of Formula IV, n is 1. In some embodiments of Formula IV, n is 2 and the first glycoside is 2 and the OR of the second glycoside 1b , OR 1c OR 1d is attached to the second glycoside via a single bond between any one of

[0109] In some embodiments of Formula IV, n is 3 and the first glycoside is 2 and the OR of the second glycoside 1b , OR 1c OR 1d and the second glycoside is attached to the second glycoside through a single bond between any one of the W 2 and OR of tertiary glycosides 1b , OR 1c OR 1d It is attached to the third glycoside via a single bond between any one of the

[0110] Provided herein is a method for synthesizing the peptide product described above, the method comprising the steps of: (a) coupling a compound of formula IV to a peptide; (b) optionally deprotecting the conjugated peptide of step (a).

[0111] In some embodiments, the deprotection comprises the use of a mild acid and / or a mild base treatment. In some embodiments of the method, the deprotection comprises the use of a strong acid.

[0112] In some embodiments, the method further comprises the step of chromatography, desalting the intermediate by reverse phase, high performance liquid chromatography, or ion exchange chromatography of the intermediate.

[0113] Provided herein are methods for improving the pharmaceutical and therapeutic response of a peptide, thereby improving its duration of action, bioavailability, or its stability in a formulation, the methods comprising covalently attaching a surfactant X to the peptide, wherein: X is:

[0114] [ka]

[0115] and During the ceremony, A is a hydrophobic group; and B is a hydrophilic group covalently attached to the peptide via a linker amino acid.

[0116] In some embodiments, A is a substituted or unsubstituted C-C 30 It is an alkyl chain, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group.

[0117] In some embodiments, A is a substituted or unsubstituted C-C 20 It is an alkyl chain, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group.

[0118] In some embodiments, A is substituted or unsubstituted C10 -C 20 It is an alkyl chain, a substituted or unsubstituted 1-alkoxyaryl group, or a substituted or unsubstituted 1-aralkyl group.

[0119] In some embodiments, B is a functionalized polyol.

[0120] In some embodiments, the polyol is a sugar.

[0121] In some embodiments, the saccharide is a monosaccharide, a disaccharide, or a polysaccharide.

[0122] In some embodiments, the sugars are selected from glucuronic acid, galacturonic acid, diglucuronic acid, and malturonic acid.

[0123] The synthetic methods described above are suitable for the synthesis of all compounds described herein, including those of Formula I, II, III, 2-I-1, 2-III, 2-V, 2-VI, 2-VII, 3-IA, 3-III-A, 3-III-B, or 3-V, and the compounds of Tables 1, 2, 3, and 4, provided in Figures 1-1 to 1-7, 2-1 to 2-4, 8-1 to 8-5, and 9-1 to 9-7, respectively.

[0124] Provided herein are methods for improving the pharmaceutical and therapeutic response of a peptide, thereby improving its duration of action, bioavailability, or its stability in a formulation, the methods comprising covalently attaching a surfactant X to the peptide, wherein: X is:

[0125] [ka]

[0126] and During the ceremony, A is a substituted or unsubstituted C1-C 20an alkyl chain, a substituted or unsubstituted alkoxyaryl group, or a substituted or unsubstituted aralkyl group; and B is a saccharide covalently attached to the peptide via a linker amino acid.

[0127] In some embodiments, the surfactant is a surfactant of the 1-alkylglycoside class.

[0128] In some embodiments, the hydrophilic group in the surfactant is attached to the peptide via an amide bond.

[0129] In some embodiments, the surfactant comprises the 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-hexadecyl-beta-D-diglucuronic acid, 1-tetradecyl-beta-D-diglucuronic acid, 1-dodecyl-beta-D-diglucuronic acid, 1-decyl-beta-D-diglucuronic acid, 1-octyl-beta-D-diglucuronic acid.

[0130] Provided herein are methods for treating pain in an individual in need thereof, comprising administering to the individual in need thereof a therapeutically effective amount of a peptide product described herein, or a compound of formula IV.

[0131] Also provided herein are covalently modified peptide and / or protein products comprising a hydrophilic group as described herein; and a hydrophobic group covalently attached to the hydrophilic group. In specific embodiments, the covalently modified peptide and / or protein product comprises a hydrophilic group that is a saccharide, and a C1-C 20 It contains a hydrophobic group that is an alkyl or aralkyl chain. [Brief explanation of the drawings]

[0132] [Figure 1-1]Table 1 in Figures 1-1 through 1-7 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 1 and 149-169. Additionally, Table 1 in Figures 1-1 through 1-7 provides the SEQ. ID Numbers for compounds EU-A101 through EU-A199 and EU-A600 through EU-A649, which have SEQ. ID. Nos. 2-148 and SEQ. ID. No. 645, respectively, as indicated. The compounds in Table 1 in Figures 1-1 through 1-7 and their respective SEQ. ID. Nos. shown in Table 1 in Figures 1-1 through 1-7 are incorporated herein as filed. [Figure 1-2] Table 1 in Figures 1-1 through 1-7 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 1 and 149-169. Additionally, Table 1 in Figures 1-1 through 1-7 provides the SEQ. ID Numbers for compounds EU-A101 through EU-A199 and EU-A600 through EU-A649, which have SEQ. ID. Nos. 2-148 and SEQ. ID. No. 645, respectively, as indicated. The compounds in Table 1 in Figures 1-1 through 1-7 and their respective SEQ. ID. Nos. shown in Table 1 in Figures 1-1 through 1-7 are incorporated herein as filed. [Figure 1-3] Table 1 in Figures 1-1 through 1-7 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 1 and 149-169. Additionally, Table 1 in Figures 1-1 through 1-7 provides the SEQ. ID Numbers for compounds EU-A101 through EU-A199 and EU-A600 through EU-A649, which have SEQ. ID. Nos. 2-148 and SEQ. ID. No. 645, respectively, as indicated. The compounds in Table 1 in Figures 1-1 through 1-7 and their respective SEQ. ID. Nos. shown in Table 1 in Figures 1-1 through 1-7 are incorporated herein as filed. [Figure 1-4]Table 1 in Figures 1-1 through 1-7 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 1 and 149-169. Additionally, Table 1 in Figures 1-1 through 1-7 provides the SEQ. ID Numbers for compounds EU-A101 through EU-A199 and EU-A600 through EU-A649, which have SEQ. ID. Nos. 2-148 and SEQ. ID. No. 645, respectively, as indicated. The compounds in Table 1 in Figures 1-1 through 1-7 and their respective SEQ. ID. Nos. shown in Table 1 in Figures 1-1 through 1-7 are incorporated herein as filed. [Figure 1-5] Table 1 in Figures 1-1 through 1-7 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 1 and 149-169. Additionally, Table 1 in Figures 1-1 through 1-7 provides the SEQ. ID Numbers for compounds EU-A101 through EU-A199 and EU-A600 through EU-A649, which have SEQ. ID. Nos. 2-148 and SEQ. ID. No. 645, respectively, as indicated. The compounds in Table 1 in Figures 1-1 through 1-7 and their respective SEQ. ID. Nos. shown in Table 1 in Figures 1-1 through 1-7 are incorporated herein as filed. [Figure 1-6] Table 1 in Figures 1-1 through 1-7 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 1 and 149-169. Additionally, Table 1 in Figures 1-1 through 1-7 provides the SEQ. ID Numbers for compounds EU-A101 through EU-A199 and EU-A600 through EU-A649, which have SEQ. ID. Nos. 2-148 and SEQ. ID. No. 645, respectively, as indicated. The compounds in Table 1 in Figures 1-1 through 1-7 and their respective SEQ. ID. Nos. shown in Table 1 in Figures 1-1 through 1-7 are incorporated herein as filed. [Figure 1-7]Table 1 in Figures 1-1 through 1-7 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 1 and 149-169. Additionally, Table 1 in Figures 1-1 through 1-7 provides the SEQ. ID Numbers for compounds EU-A101 through EU-A199 and EU-A600 through EU-A649, which have SEQ. ID. Nos. 2-148 and SEQ. ID. No. 645, respectively, as indicated. The compounds in Table 1 in Figures 1-1 through 1-7 and their respective SEQ. ID. Nos. shown in Table 1 in Figures 1-1 through 1-7 are incorporated herein as filed. [Figure 2-1] Table 2 in Figures 2-1 through 2-4 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 170-174 and SEQ. ID. Nos. 283-302. Additionally, Table 2 in Figures 2-1 through 2-4 provides the SEQ. ID Numbers for compounds EU-201 through EU-299 and EU-900 through EU-908, which have SEQ. ID. Nos. 175-282, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 2 in Figures 2-1 through 2-4 are incorporated herein as filed. [Figure 2-2] Table 2 in Figures 2-1 through 2-4 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 170-174 and SEQ. ID. Nos. 283-302. Additionally, Table 2 in Figures 2-1 through 2-4 provides the SEQ. ID Numbers for compounds EU-201 through EU-299 and EU-900 through EU-908, which have SEQ. ID. Nos. 175-282, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 2 in Figures 2-1 through 2-4 are incorporated herein as filed. [Figure 2-3]Table 2 in Figures 2-1 through 2-4 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 170-174 and SEQ. ID. Nos. 283-302. Additionally, Table 2 in Figures 2-1 through 2-4 provides the SEQ. ID Numbers for compounds EU-201 through EU-299 and EU-900 through EU-908, which have SEQ. ID. Nos. 175-282, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 2 in Figures 2-1 through 2-4 are incorporated herein as filed. [Figure 2-4] Table 2 in Figures 2-1 through 2-4 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 170-174 and SEQ. ID. Nos. 283-302. Additionally, Table 2 in Figures 2-1 through 2-4 provides the SEQ. ID Numbers for compounds EU-201 through EU-299 and EU-900 through EU-908, which have SEQ. ID. Nos. 175-282, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 2 in Figures 2-1 through 2-4 are incorporated herein as filed. [Figure 3] Figure 3 has two panels. The upper panel in Figure 3 illustrates the interaction between PTH1-34 or PTHrP1-34 and the PTH R1 receptor based on the X-ray crystal structure of the receptor's extracellular domain (Piozak, AA, et al. (2009) J Biol Chem 284:28382-391). The key hydrophobic interactions of residues 23' (W or F), 24' (L), and 28' (L or I) are illustrated. In some instances, surfactant modifications on the modified peptides described herein replace these key hydrophobic interactions. The lower panel in Figure 3 shows a structural comparison between the sequences of PTH1-34 and PTHrP1-34, illustrating strong regions of identity and homology in sequence and peptide helical structure. [Figure 4] Figure 4 shows the cAMP response of human cells in culture (SaOS2) to stimulation with EU-232, a representative peptide product described herein. The vertical axis shows the response as a percentage of the maximal response exhibited by the internal assay standard, i.e., PTHrP. The data demonstrate a super-agonistic response to EU-232. [Figure 5] Figure 5 shows the response of human cells in culture (SaOS2) to treatment with various doses of EU-285 (a coded sample of human PTHrP). The ordinate (vertical axis) shows the cAMP response as a percentage of the maximal response of the internal assay standard, i.e., PTHrP. [Figure 6] Figure 6 shows blood phosphate levels in rat serum tested at various time points after subcutaneous dosing of rats with saline (G1), 80 micrograms per kg PTH (G2), 80 micrograms per kg EU-232 (G3), or 320 micrograms per kg EU-232 (G4). This surfactant-modified analog, EU-232, demonstrates an extended duration of action as evidenced by the maximal statistically significant effect seen at the last time point in the assay (i.e., 5 hours after dosing). See Example 6. [Figure 7] Figure 7 shows blood calcium levels in rat serum at various time points after subcutaneous administration of saline (G1), 80 micrograms per kg of PTH (G2), 80 micrograms per kg of EU-232 (G3), or 320 micrograms per kg of EU-232 (G4). None of the groups were statistically significantly different from the control (G1). Furthermore, the maximally effective dose and time point for EU-232 (G4; at 5 hours) showed no elevation, and therefore no trend toward hypercalcemia at the maximally effective dose was observed. See Example 6. [Figure 8-1]Table 3 in Figures 8-1 through 8-5 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 3 in Figures 8-1 through 8-5 provides the SEQ. ID Numbers for compounds EU-A300 through EU-A425, which have SEQ. ID. Nos. 306-431, respectively, as shown. The compounds in Table 3 in Figures 8-1 through 8-5 and their respective SEQ. ID. Nos. shown in Table 3 in Figures 8-1 through 8-5 are incorporated herein as filed. [Figure 8-2] Table 3 in Figures 8-1 through 8-5 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 3 in Figures 8-1 through 8-5 provides the SEQ. ID Numbers for compounds EU-A300 through EU-A425, which have SEQ. ID. Nos. 306-431, respectively, as shown. The compounds in Table 3 in Figures 8-1 through 8-5 and their respective SEQ. ID. Nos. shown in Table 3 in Figures 8-1 through 8-5 are incorporated herein as filed. [Figure 8-3] Table 3 in Figures 8-1 through 8-5 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 3 in Figures 8-1 through 8-5 provides the SEQ. ID Numbers for compounds EU-A300 through EU-A425, which have SEQ. ID. Nos. 306-431, respectively, as shown. The compounds in Table 3 in Figures 8-1 through 8-5 and their respective SEQ. ID. Nos. shown in Table 3 in Figures 8-1 through 8-5 are incorporated herein as filed. [Figure 8-4]Table 3 in Figures 8-1 through 8-5 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 3 in Figures 8-1 through 8-5 provides the SEQ. ID Numbers for compounds EU-A300 through EU-A425, which have SEQ. ID. Nos. 306-431, respectively, as shown. The compounds in Table 3 in Figures 8-1 through 8-5 and their respective SEQ. ID. Nos. shown in Table 3 in Figures 8-1 through 8-5 are incorporated herein as filed. [Figure 8-5] Table 3 in Figures 8-1 through 8-5 shows compounds prepared by the methods described herein. This specification provides the sequences for SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 3 in Figures 8-1 through 8-5 provides the SEQ. ID Numbers for compounds EU-A300 through EU-A425, which have SEQ. ID. Nos. 306-431, respectively, as shown. The compounds in Table 3 in Figures 8-1 through 8-5 and their respective SEQ. ID. Nos. shown in Table 3 in Figures 8-1 through 8-5 are incorporated herein as filed. [Figure 9-1] Table 4 in Figures 9-1 through 9-7 lists compounds prepared by the methods described herein. The present specification provides SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 4 in Figures 9-1 through 9-7 provides SEQ. ID Numbers for compounds EU-A426 through EU-599, which have SEQ. ID. Nos. 432-520, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 4 in Figures 9-1 through 9-7 are incorporated herein as filed. [Figure 9-2]Table 4 in Figures 9-1 through 9-7 lists compounds prepared by the methods described herein. The present specification provides SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 4 in Figures 9-1 through 9-7 provides SEQ. ID Numbers for compounds EU-A426 through EU-599, which have SEQ. ID. Nos. 432-520, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 4 in Figures 9-1 through 9-7 are incorporated herein as filed. [Figure 9-3] Table 4 in Figures 9-1 through 9-7 lists compounds prepared by the methods described herein. The present specification provides SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 4 in Figures 9-1 through 9-7 provides SEQ. ID Numbers for compounds EU-A426 through EU-599, which have SEQ. ID. Nos. 432-520, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 4 in Figures 9-1 through 9-7 are incorporated herein as filed. [Figure 9-4] Table 4 in Figures 9-1 through 9-7 lists compounds prepared by the methods described herein. The present specification provides SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 4 in Figures 9-1 through 9-7 provides SEQ. ID Numbers for compounds EU-A426 through EU-599, which have SEQ. ID. Nos. 432-520, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 4 in Figures 9-1 through 9-7 are incorporated herein as filed. [Figure 9-5]Table 4 in Figures 9-1 through 9-7 lists compounds prepared by the methods described herein. The present specification provides SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 4 in Figures 9-1 through 9-7 provides SEQ. ID Numbers for compounds EU-A426 through EU-599, which have SEQ. ID. Nos. 432-520, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 4 in Figures 9-1 through 9-7 are incorporated herein as filed. [Figure 9-6] Table 4 in Figures 9-1 through 9-7 lists compounds prepared by the methods described herein. The present specification provides SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 4 in Figures 9-1 through 9-7 provides SEQ. ID Numbers for compounds EU-A426 through EU-599, which have SEQ. ID. Nos. 432-520, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 4 in Figures 9-1 through 9-7 are incorporated herein as filed. [Figure 9-7] Table 4 in Figures 9-1 through 9-7 lists compounds prepared by the methods described herein. The present specification provides SEQ. ID. Nos. 303-305 and SEQ. ID. Nos. 619-644. Additionally, Table 4 in Figures 9-1 through 9-7 provides SEQ. ID Numbers for compounds EU-A426 through EU-599, which have SEQ. ID. Nos. 432-520, respectively, as indicated. The compounds in Table 2 in Figures 2-1 through 2-4 and their respective SEQ. ID. Nos. shown in Table 4 in Figures 9-1 through 9-7 are incorporated herein as filed. [Figure 10]Figure 10 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), showing the receptor and the key hydrophobic binding elements of the ligand exendin-4 (Val19*, Phe22*, Trp25*, Leu26*) that are mimicked and replaced by the hydrophobic 1'-alkyl moiety of the surfactant on the peptides of the invention. [Figure 11] Figure 11 shows cell assay data for a representative compound, EU-A178, which acts on cells containing mu opioid receptors (MOP) and competes with 30 nM DPDPE to act as an antagonist at delta opioid receptors (DOP). The representative compound exhibits potent, fully antagonistic behavior in the MOP agonist assay and highly potent activity as a pure antagonist at the DOP receptor. DETAILED DESCRIPTION OF THE INVENTION

[0133] Provided herein are modified peptides and / or proteins, including peptides and / or proteins covalently attached to a hydrophilic group, "head" (e.g., a polyol (e.g., a sugar)); the hydrophilic group is covalently attached to a hydrophobic group, "tail," thereby generating a surfactant. In some embodiments, the use of hydrophobically linked glycoside surfactant (e.g., alkyl glycoside) moieties for covalent modification of peptides or proteins extends the duration of action of the peptide or protein through multiple mechanisms, including the formation of drug depots at the site of administration in the body and binding to hydrophobic carrier proteins. In some embodiments, the incorporation of steric hindrance into 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 attachment of alkyl glycoside class of surfactants) increases transport across mucosal barriers. Thus, the peptide and / or protein modifications described herein provide desirable benefits, including, but not limited to, protection from proteolysis and delayed movement from the site of administration, thereby leading to sustained pharmacokinetic behavior (e.g., prolonged circulating t1 / 2) and improved oral mucosal bioavailability.

[0134] In some embodiments, the interactions of improved peptides and / or proteins with their receptors are modified in beneficial ways by sequence truncation, introduction of constraints, and / or incorporation of steric hindrance. Described herein are new alkyl glycoside reagents that allow for the incorporation of both rigidity and steric hindrance in the modified peptides and / or proteins. In some embodiments, steric hindrance confers receptor selectivity for the modified peptides and / or proteins described herein. In some embodiments, steric hindrance provides protection from proteolysis.

[0135] Proteins and peptides undergo numerous physical and chemical changes that can affect efficacy and safety. Among these is aggregation, which includes dimerization, trimerization, and the formation of higher-order aggregates such as amyloids. Aggregation is a major problem underlying several potentially adverse effects on peptide and / or protein-based therapeutics, including loss of efficacy, altered pharmacokinetics, reduced stability or shelf life of the product, and the induction of undesirable immunogenicity. The bioavailability and pharmacokinetics of self-associating peptides can be affected by the size of the aggregate and the ease of disruption of noncovalent intermolecular interactions at subcutaneous sites (Maji, SK, et al. (2008) PLoS Biol 6: e17). In some instances, peptides can aggregate into subcutaneous depots and disassociate with a t1 / 2 of 30 days or more. Such slow degradation can lead to desirable effects, such as one-month delivery from a single subcutaneous injection, causing low blood concentrations such that the peptide appears inactive in vivo. Thus, hydrophobic aggregation appears to completely eliminate peptide bioavailability and efficacy (Clodfelter, DK, et al. (1998) Pharm Res 15: 254-262).

[0136] Aggregation has been associated with increased immunogenicity of administered peptide and / or protein therapeutics. One way to circumvent this problem is to work with lower concentration solutions, but concentrated peptide and protein solutions are desirable in some instances for ease of administration. In some instances, adding polyols (e.g., mono- or oligosaccharides) or alkyl glycosides to the peptide and / or protein solution during purification and concentration reduces or eliminates aggregation, providing greater efficacy during the manufacturing process and providing a final product that is less likely to be immunogenic.

[0137] The FDA and other regulatory agencies are increasingly scrutinizing aggregation, particularly because of its potential link to undesirable immunogenicity. The immunogenicity of self-associating peptides can be affected by the formation of aggregates as a result of non-covalent intermolecular interactions. For example, interferon has been shown to aggregate, resulting in an antibody response (Hermeling, S., et al. (2006) J Pharm Sci 95: 1084-1096). Antibody responses to erythropoietin have led to "erythrocyte aplasia," a potentially life-threatening side effect, in many patients receiving recombinant EPO (Casadevall, N., et al. (2002) N Engl J Med 346: 469-475) after formulation changes that altered the source and concentration of serum albumin. Insulin loses activity due to protein aggregation after agitation at temperatures exceeding those found in refrigerated storage (Pezron, I., et al. (2002) J Pharm Sci 91: 1135-1146, Sluzky, V., et al. (1991) Proc Natl Acad Sci USA 88: 9377-9381). Monoclonal antibody-based therapeutics are prone to inactivation as a result of protein aggregation (King, HD, et al. (2002) J Med Chem 45: 4336-4343). Highly concentrated monoclonal antibody formulations pose stability, manufacturing, and delivery challenges related to the potential for aggregation of these antibodies. Enzymes can also lose activity as a result of aggregation. For example, heat inactivation of urokinase has been reported to occur via aggregation (Porter, WR, et al. (1993) Thromb Res 71: 265-279).

[0138] Stabilizing proteins during lyophilization has also posed problems. Protein therapeutics often lose biological activity after lyophilization and reconstitution as a result of aggregate formation and precipitation. In some cases, aggregation is reduced by adding reconstitution additives (e.g., polysaccharides, polyphosphates, amino acids, polyethylene glycol (PEG), and various surfactants) (Zhang, MZ, et al. (1995) Pharm Res 12: 1447-1452, Vrkljan, M., et al. (1994) Pharm Res 11: 1004-1008). In some cases, alcohols or combinations of other organic solvents are used for solubilization. Trifluoroethanol is effective in maintaining peptide structure and has been used in mixtures to stabilize various peptides (Roccatano, D., et al. (2002) Proc Natl Acad Sci USA 99: 12179-12184). Such agents can have harsh effects on mucosal tissue, potentially causing patient discomfort or local toxic effects. U.S. Patent Nos. 7,390,788 and 7,425,542 describe the use of alkyl glycosides as stabilizers due to their gentle, nonionic surfactant-like effect. However, the covalent incorporation of alkyl glycosides into peptide and / or protein structures themselves has not previously been described.

[0139] Frequently, naturally occurring oligosaccharides that are covalently attached to proteins do not have surfactant characteristics. In some embodiments, the peptide and / or protein products described herein have covalently attached saccharides and additional hydrophobic groups that confer surfactant characteristics to the modified peptide, thereby allowing tuning of the bioavailability, immunogenicity, and / or pharmacokinetic behavior of surfactant-modified peptides.

[0140] Oligosaccharide-modified proteins and peptides have been synthesized by enzymatic incorporation of sugars or oligosaccharides (Gijsen, HJ, 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 chemical methods (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), as described, for example, in Jensen, KJ and Brask, J. (2005) Biopolymers 80: 747-761. Peptides as well as proteins have 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-Pilard, 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, S.A., et al. (2001) Org Lett 3:3723-3725; Pean, C., et al. (2001) Biochim Biophys Acta 1541:150-160; Filira, F., et al. (2003) Org Biomol Chem 1:3059-3063; Grotenbreg, G.M., 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; Lowery JJ, et al. (2011) J Pharmacol Exptl Therap 336: 767-78; Yamamoto, T., et al. (2009) J Med Chem 52: 5164-5175)。.

[0141] However, the aforementioned attempts do not describe additional hydrophobic groups attached to the oligosaccharides linked to the peptide. Accordingly, provided herein are modified peptides and / or proteins incorporating hydrophobic groups attached to saccharides and / or oligosaccharides that are covalently attached to the peptides and / or proteins, allowing for tuning of bioavailability, immunogenicity, and pharmacokinetic behavior. Accordingly, also provided herein are surfactant reagents comprising oligosaccharides and hydrophobic groups, which allow for modification of peptides and / or proteins.

[0142] Provided herein is the use of saccharide-based surfactants in covalent attachment to peptides to improve the properties of the peptides and / or proteins. In some embodiments, surfactant modification of peptides and / or proteins as described herein (e.g., covalent attachment of alkylglycoside class of surfactants) increases transport across mucosal barriers. In some embodiments, covalent attachment of surfactants to peptide and / or protein products prevents aggregation of the peptide and / or protein.

[0143] The surfactant-modified peptides and / or proteins described herein overcome limitations of peptide formulations, including but not limited to, short duration of action, poor bioavailability, aggregation, immunogenicity, and lack of receptor subtype specificity, through the covalent incorporation of surfactants such as alkylglycosides as novel peptide and protein modifiers.

[0144] In certain instances, the effects of surfactants are beneficial to the physical properties or efficacy of pharmaceutical formulations, but can be irritating to skin and / or other tissues, particularly mucous membranes, such as those found in the nose, mouth, eyes, vagina, rectum, buccal, or sublingual areas. Furthermore, in some instances, surfactants denature proteins, disrupting their biological function. Because surfactants exert their effects on the critical micelle concentration (CMC), surfactants with low CMCs are desirable so that they can be effectively utilized at low concentrations or in small amounts in pharmaceutical formulations. Thus, in some embodiments, surfactants (e.g., alkyl glycosides) suitable for peptide modification as described herein have a CMC of less than about 1 mM in pure or aqueous solutions. By way of example only, specific CMC values ​​for alkylglycosides in water are: octylmaltoside 19.5 mM; decylmaltoside 1.8 mM; dodecyl-β-D-maltoside 0.17 mM; tridecylmaltoside 0.03 mM; tetradecylmaltoside 0.01 mM; and sucrose dodecanoate 0.3 mM. Naturally, suitable surfactants may have higher or lower CMCs depending on the peptide and / or protein being modified. As used herein, "critical micelle concentration" or "CMC" refers to the concentration of an amphiphilic component (an alkylglycoside) in a solution at which micelles (such as spherical micelles, rounded rods, or lamellar structures) begin to form in the solution. In certain embodiments, the alkyl glycosides dodecyl, tridecyl, and tetradecyl maltoside or glucoside, as well as sucrose dodecanoate, tridecanoate, and tetradecanoate, have lower CMCs and are suitable for the peptide and / or protein modifications described herein.

[0145] Opioid peptides and analogs In some embodiments, a class of peptide therapeutics applicable to the peptide modification methods described herein is the class of peptide opioids. This class is derived from endogenous peptide opioids, which have a wide range of functions in the body, carried out through binding to mu (MOR), delta (DOR), and kappa (KOR) opioid receptors (Schiller, PW (2005) AAPS J 7: E560-565). Their role in most subjects is to modify pain signals, particularly to inhibit the transmission and perception of pain signals. In developing such drugs, central side effects (respiratory depression, place preference indicating reward from self-administration) are a major problem, and therefore peripherally acting drugs would be attractive (Stein, C., et al. (2009) Brain Res Rev 60: 90-113). Research from many laboratories has suggested that the optimal class of drugs would have mu opioid receptor agonism with delta receptor antagonism potential (Schiller, PW (2010) Life Sci 86: 598-603).

[0146] Endomorphins (Janecka, A., et al. (2007) Curr Med Chem 14: 3201-3208) are primarily specific for the mu receptor, but judicious framework modifications can result in molecules with both mu and delta selectivity (Lazarus, LH and Okada, Y. (2012) Expert Opin Ther Patents 22: 1-14; Keresztes, A., et al. (2010) ChemMedChem 5: 1176-96). Derived from the dermorphin family are the DALDA class of analogs (Schiller, PW (2010) Life Sci 86: 598-603). Also derived from the dermorphin family are the TIPP family of peptides (Schiller, PW, et al., (1999) Biopolymers 51:411-25). Described herein are certain opioid peptides that are covalently attached to the saccharide groups of alkylglycoside surfactants and have improved pharmaceutical properties.

[0147] Exemplary synthetic peptide analogs described herein are derived from endomorphins and dermorphins, two classes of naturally occurring peptide opioid sequences. In one embodiment, peptide analogs of the present invention of naturally occurring sequences are endorphin-related sequences, such as those exemplified by EU-A101 through EU-A115. In another embodiment, the peptide analogs are dermorphin-related sequences, such as EU-A107, EU-A108, and EU-A120, through EU-A133. A related class of opioid peptide analogs is exemplified by the sequences EU-A134 through EU-A142, in which a Tic residue replaces the D-Ala residue found in the dermorphin structure (TIPP family). A further class of specialized binding is exhibited when Tic is exchanged for Tic(Ψ[CH2-NH]), since it requires the complementary residue Ψ-Phe to complete binding.

[0148] In some embodiments, the surfactant-modified peptide product has an amino acid sequence corresponding to the following general formula III:

[0149] [ka]

[0150] During the ceremony, aa1 is Tyr, Dmt, N-alkyl-Tyr, N-alkyl-Dmt, N-dialkyl-Tyr, N-dialkyl-Dmt, etc.; aa2 is Pro, D-Arg, DU(X), D-Ala, Tic, Tic(Ψ[CH2-NH]); aa3 is Phe, Typ, Tmp, D- or L-Nal(1), D- or L-Nal(2), CαMePhe, Ψ-Phe; aa4 is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), U(X), D- or L-CαMeU(X); aa5 is absent, Pro, Aib, U(X), D- or L-CαMeU(X), Alkyl or dialkyl is independently a substituted or unsubstituted C-C 10 a branched or straight chain, or a substituted or unsubstituted aralkyl chain of U is the binding amino acid; X is a functionalized surfactant attached to the side chain of U; Z is —OH or NH 2 .

[0151] In some specific embodiments of Formula III, X has the structure:

[0152] [ka]

[0153] During the ceremony: A is a substituted or unsubstituted C1-C 30is 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 is a single bond.

[0154] In one embodiment, the N-alkyl is N-methyl; U is a dibasic amino acid, such as Lys or Orn; where X is a modified nonionic surfactant of the 1-alkylglycoside class, where alkyl is C1-C 20 alkyl or alkoxyaryl substituents, and the glycosidic bond to the sugar ring is via -O- or other heteroatom (e.g., S or N); Z is NH2.

[0155] In another embodiment, the 1-alkyl group in the 1-alkyl-glycoside is a substituted or unsubstituted C-C 16 is alkyl; U is Lys and Z is NH2; aa1 is Tyr, Dmt, Nα-Me-Tyr, Nα-Me-Dmt; aa2 is Pro; aa3 is Phe, Trp, Tmp; aa4 is Phe, Lys(X); aa5 is absent or Lys(X).

[0156] In a further embodiment, the 1-alkyl group in the 1-alkyl glycoside is a substituted or unsubstituted C-C 20 alkyl; Z is NH; aa1 is Tyr, Dmt; aa2 is D-Arg, D-Lys(X), Tic, Tic(Ψ[CH2-NH]); aa3 is Phe, Trp, Tmp, CαMePhe, Ψ-Phe; aa4 is Phe, Tmp, Lys(X); aa5 is absent, Pro, Aib, Lys(X), D- or L-CαMeLys(X).

[0157] The parent structure of the endomorphin class is: Endomorphin 1 - Tyr-Pro-Trp-Phe-NH2 Endomorphin 2 - Tyr-Pro-Phe-Phe-NH2

[0158] Substituents for particular analog families are shown below:

[0159] [Table 1]

[0160] The specific sequence contains the following amino acids:

[0161] [Table 2]

[0162] In specific embodiments, analogs having attached surfactants include, but are not limited to:

[0163] [Table 3]

[0164] The parent structure of the dermorphin class is: Parent dermorphin - Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2.

[0165] Specific analog family configurations are shown below:

[0166] [Table 4]

[0167] In some embodiments, specific analogs suitable for surfactant attachment include, but are not limited to:

[0168] [Table 5]

[0169] In specific embodiments, analogs suitable for surfactant attachment include, but are not limited to:

[0170] [Table 6-1]

[0171] [Table 6-2]

[0172] Within the scope of the embodiments presented herein, peptide chains substituted at appropriate positions are contemplated by substitution of saturated or unsaturated alkyl chains of the analogs claimed herein with fatty acids such as octanoic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, 3-phenylpropanoic acid, for example, by acylation on the linker amino acid at the ε-position of Lys (Nestor, JJ, Jr. (2009) Current Medicinal Chemistry 16: 4399-4418; Zhang, L and Bulaj, G. (2012) Curr Med Chem 19: 1602-18). Non-limiting, illustrative examples of such analogs are as follows:

[0173] H-Dmt-Tic-Phe-Lys(N-epsilon-acetyl)-NH2, (SEQ.ID.NO.161).

[0174] H-Dmt-Tic-Phe-Lys(N-epsilon-dodecanoyl)-NH2, (SEQ.ID.NO.162).

[0175] H-Dmt-Tic-Phe-Lys(N-epsilon-tetradecanoyl)-NH2, (SEQ.ID.NO.163).

[0176] H-Dmt-Tic-Phe-Lys(N-epsilon-(gamma-glutamyl)-N-alpha-dodecanoyl))-NH2, (SEQ.ID.NO.164).

[0177] H-Dmt-Tic-Phe-Lys(N-epsilon-(gamma-glutamyl)-N-alpha-tetradecanoyl))-NH2, (SEQ.ID.NO.165).

[0178] H-Dmt-Tic-Phe-Lys(N-epsilon-acetyl)-NH-benzyl, (SEQ.ID.NO.166).

[0179] H-Dmt-Tic-Phe-Lys(N-epsilon-dodecanoyl)-NH-benzyl, (SEQ.ID.NO.167).

[0180] In other embodiments of the present invention, the peptide chain can be substituted at appropriate positions by reaction of a linker amino acid, e.g., a sulfhydryl of Cys, with a spacer and a 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. Non-limiting examples of such molecules include:

[0181] H-Dmt-Tic-Phe-Cys(S-(3-(PEG4-aminoethylacetamido-cholesterol)))-NH2, (SEQ.ID.NO.168).

[0182] H-Dmt-Tic-Phe-Cys(S-(3-(PEG4-aminoethylacetamido-cholesterol)))-NH-benzyl, (SEQ. ID. NO. 169) and the like.

[0183] Compounds of Formula I, Formula II, or Formula III are assayed for mu opioid receptor activity (MOP in agonist and antagonist modes) in a cellular assay, and for Delta 2 opioid receptor activity (DOP in agonist and antagonist modes) in a cellular assay as described in Example 12.

[0184] In certain embodiments, compounds with pure MOP agonist activity along with pure DOP antagonist activity are suitable properties for clinical application, as shown in Example 12. Also contemplated within the scope of this disclosure are compounds that have low solubility and low apparent in vitro potency, but exhibit an extended duration of action in vivo.

[0185] Within the embodiments presented herein, peptide products of Formula I, Formula II, or Formula III are contemplated, wherein the peptide product includes one or more surfactant groups (e.g., a group X having the structure of Formula I). ​​In one embodiment, the peptide product of Formula I, Formula II, or Formula III includes one surfactant group. In another embodiment, the peptide product of Formula I, Formula II, or Formula III includes two surfactant groups. In yet another embodiment, the peptide product of Formula I, Formula II, or Formula III includes three surfactant groups.

[0186] <PTHペプチドおよびアナログ> Provided herein, in some embodiments, are reagents and intermediates for the synthesis of modified peptides and / or proteins (eg, modified PTH, PTHrP, etc.) through the incorporation of surfactants.

[0187] Provided herein, in some embodiments, is a peptide product comprising a surfactant X covalently attached to a peptide, the peptide comprising a linker amino acid U and at least one other amino acid:

[0188] [ka]

[0189] wherein surfactant X is a group of formula 2-I:

[0190] [ka]

[0191] During the ceremony: R 1a is independently in each occurrence a single bond, H, or a substituted or unsubstituted C-C 30 an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a moiety containing a steroid nucleus; R 1b , R 1c , and R 1d is independently in each occurrence a single bond, H, or a substituted or unsubstituted C-C 30 an alkyl, 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 in each occurrence H, substituted or unsubstituted C-C 30Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, -NH2, -SH, C2-C4-alkene, C2-C4-alkyne, -NH(C=O)-CH2-Br, -(CH2) m -maleimide, or -N3; n is 1, 2, or 3; and m is 1-10; The peptide is selected from Formula 2-II:

[0192] [ka]

[0193] During the ceremony: Z is OH or -NH-R 3 and R 3 is H, substituted or unsubstituted C1-C 12 alkyl, or a PEG chain of less than 10 Da; aa1 is Aib, Ac5c, or Deg; aa3 is Aib, Ac4c, or Deg; aa5 is His, or Ile; aa6 is Gln or Cit; aa7 is Leu or Phe; aa8 is Leu, or Nle; aa 10 is Asp, Asn, Gln, Glu, Cit, Ala, or Aib; aa 11 is Arg, or hArg; aa 12 is glycine, glutamic acid, rees, ala, Aib or Ac5c; aa 13 is Lys, or Arg; aa 14 is Ser, His, Trp, Phe, Leu, Arg, Lys, Glu, or NaI(2); aa 15is Ile, Leu, or Aib; aa 16 is Gln, Asn, Glu, Lys, Ser, Cit, Aib, or U; aa 17 is Asp, Ser, Aib, Ac4c, Ac5c or U; aa 18 is absent or is Leu, Gln, Cit, Aib, Ac5c, Lys, Glu, or U; aa 19 is absent or is Arg, Gln, Aib, Ac4c, Ac5c, or U; aa 20 is absent or is Arg, Gln, Lys, Aib, Ac4c, Ac5c, or U; aa 21 is absent or is Arg, Val, Aib, Ac5C, Deg, or U; aa 22 is absent or is Phe, Glu, Aib, Ac5C, Lys, or U; aa 23 is absent or is Leu, Phe, Trp, or U; aa 24 is absent or is His, Arg, or U; aa 25 is absent or is His, Lys, or U, and aa 26 is absent or Aib, Ac5c, Lys; U is a natural or unnatural amino acid containing a functional group used to covalently attach to surfactant X; where aa1-aa 26 any two of which are optionally cyclized via their side chains to form a lactam bond; However, aa 16 -aa 26 is a linker amino acid U covalently attached to X.

[0194] In some embodiments, n is 1. In some embodiments, n is 2 and the first glycoside is a hydroxyl group of the first glycoside. 2 and the OR of the second glycoside 1b , OR 1c OR 1d is attached to the second glycoside via a single bond between any one of

[0195] In some embodiments, n is 3 and the first glycoside is 2 and the OR of the second glycoside 1b , OR 1c OR 1d and the second glycoside is attached to the second glycoside through a single bond between any one of the W 2 and OR of tertiary glycosides 1b , OR 1c OR 1d It is attached to the third glycoside via a single bond between any one of the

[0196] In one embodiment compound of formula IA is a compound where X has the structure:

[0197] [ka]

[0198] During the ceremony: R 1a is H, a protecting group, a substituted or unsubstituted C1-C 30 an alkyl group or a moiety containing a steroid nucleus; R 1b , R 1c , and R 1d are each independently at each occurrence H, a protecting group, or a substituted or unsubstituted C-C 30 is an alkyl 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-, or -S-; R 2 is a single bond, a C2-C4 alkene, a C2-C4 alkyne, or -(CH2) m -maleimide; and m is 1-10.

[0199] In another embodiment, the compound of formula IA is a compound where X has the structure:

[0200] [ka]

[0201] Thus, in the embodiments described above, R 2 is a single bond.

[0202] For example, in exemplary embodiments of the structure of X described above, W 1 is -C(=O)NH- and R 2 is W 1 and an amino acid residue U in the peptide (for example, the amino group in the side chain of a lysine residue present in the peptide).

[0203] In a further embodiment, the compound of formula IA is one in which X has the structure:

[0204] [ka]

[0205] For example, in exemplary embodiments of the structure of X described above, W 1 is -CH2- and R 2 is the alkyl-linked maleimide functional group on X, and R 2 is attached to the appropriate position of an amino acid residue U in the peptide (for example, the thiol group in a cysteine ​​residue of the peptide forms a thioether with the maleimide on X).

[0206] In yet another embodiment, the compound of formula IA is a compound where X has the structure:

[0207] [ka]

[0208] During the ceremony: R 1a is H, a protecting group, a substituted or unsubstituted C1-C 30 an alkyl group or a moiety containing a steroid nucleus; R 1b , R 1c , and R 1d are each independently at each occurrence H, a protecting group, or a substituted or unsubstituted C-C 30 is an alkyl group; W 1 is —(C═O)—NH—; W 2 is -O-; and R 2 is a single bond.

[0209] In a further embodiment, the compound of formula IA is a compound where X has the structure:

[0210] [ka]

[0211] During the ceremony: R 1a is a substituted or unsubstituted C1-C 30 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 is a single bond.

[0212] In some embodiments described above or herein, R 1a is a substituted or unsubstituted C1-C 30 It is an alkyl group.

[0213] In some embodiments described above or herein, R 1a is a substituted or unsubstituted C6-C 20 It is an alkyl group.

[0214] Also contemplated herein is an alternative embodiment wherein X in formula 2-IA has the following structure:

[0215] [ka]

[0216] For example, in exemplary embodiments of the structure of X described above, W 1 is -S- and R 2 is C1-C 30 is an alkyl group, and W 2 is S and R 1a is W 2 and the appropriate portion of an amino acid residue U in the peptide (for example, a thiol group in a cysteine ​​residue of the peptide forms a thioether with X).

[0217] In another exemplary alternating embodiment of the structure of X described above, W 1 is -S- and R 2 is C1-C 30 is an alkyl group, and W 2 is O and R 1a is W 2 and the appropriate portion of the amino acid residue U in the peptide (for example, the hydroxyl group in a serine or threonine residue of the peptide forms an ether with X).

[0218] In some embodiments, U is used to covalently attach X and is a dibasic natural or unnatural amino acid, a thiol-containing natural or unnatural amino acid, an unnatural amino acid containing an -N group, an acetylene-containing unnatural amino acid, or an unnatural amino acid containing -NH-C(=O)-CH-Br or -(CH)-maleimide, where m is 1-10.

[0219] In some embodiments of the peptide product, the surfactant is a surfactant of the 1-alkylglycoside class. In some embodiments of the peptide product, the surfactant is attached to the peptide via an amide bond.

[0220] 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, 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-hexadecyl beta-D-diglucuronic acid, 1-tetradecyl beta-D-diglucuronic acid, 1-dodecyl beta-D-diglucuronic acid, 1-decyl beta-D-diglucuronic acid, 1-octyl beta-D-diglucuronic acid, or functionalized 1-eicosyl beta-D-glucose, 1-octadecyl beta-D-glucose, 1-hexadecyl beta-D-g ... -tetradecyl beta-D-glucose, 1-dodecyl beta-D-glucose, 1-decyl beta-D-glucose, 1-octyl beta-D-glucose, 1-eicosyl beta-D-maltoside, 1-octadecyl beta-D-maltoside, 1-hexadecyl beta-D-maltoside, 1-dodecyl beta-D-maltoside, 1-decyl beta-D-maltoside, 1-octyl beta-D-maltoside, etc., and the peptide product is prepared by forming a bond between the aforementioned group and a group on the peptide (e.g., a —COOH group on the aforementioned group and an amino group on the peptide).

[0221] In some embodiments of the peptide product, U is the 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 naturally occurring D- or L-amino acid. In some embodiments of the peptide product, U is an unnatural amino acid. In some embodiments of the peptide product, U is selected from Lys, Cys, Orn, or an unnatural amino acid that includes a functional group that is used to covalently attach to surfactant X.

[0222] In some embodiments of the peptide product, the functional group used to covalently attach the peptide to surfactant X is -NH, -SH, -OH, -N, haloacetyl, -(CH) m -maleimide (where m is 1-10), or an acetylene group.

[0223] In some embodiments, the side chain functional groups of two different amino acid residues are linked to form a cyclic lactam. For example, Lys 14 The side chain is Glu 18 can form a cyclic lactam with the side chain of Lys 18 is Glu 22 In some embodiments, such lactam structures can be inverted, e.g., to form lactams with the side chain of Glu. 14 and Lys 18 Such lactam bonds, in some instances, stabilize alpha-helical structures in peptides (Condon, SM, et al. (2002) Bioorg Med Chem 10: 731-736).

[0224] In some embodiments, the peptide product containing a covalently attached alkyl glycoside is a covalently modified PTH or analog thereof. In some such embodiments, the peptide product contains a covalently attached 1-O-alkyl β-D-glucuronic acid and the peptide is an analog of PTH.

[0225] In some embodiments, the peptide product comprising a covalently attached alkyl glycoside is a covalently modified PTHrP or an analog thereof. In some such embodiments, the peptide product comprises a covalently attached 1-O-alkyl β-D-glucuronic acid, and the peptide is an analog of PTHrP.

[0226] In some embodiments, the peptide product has the structure of Formula 2-II:

[0227] [ka]

[0228] During the ceremony: Z is OH or -NH; aa1 is Aib, or Ac5c; aa 12 is Ala, Glu, Lys, Aib, or Ac5c; aa 14 is Trp, Phe, Lys, Glu, or NaI(2); aa 16 is Gln, Asn, Glu, Lys, Cit, or U(X); aa 17 is Asp, Ser, Aib, Ac4c, Ac5C, or U(X); aa 18 is absent or is Leu, Gln, Aib, Lys, Glu, or U(X); aa 19 is absent or is Arg, Glu, Aib, Ac4c, or Ac5c; aa 20 is absent or Arg, Glu, Lys, Aib, Ac4c, Ac5c; aa 21 is absent or is Arg, Val, Aib, Ac5C, or Deg; aa 22 is absent, or is Phe, Glu, Lys, or U(X). aa 23 is absent, or is Leu, Phe, Trp, or U(X). aa 24 is absent or is His, Arg, or U(X); aa 25 is absent or is His, Lys, or U(X); and aa 26 is absent or Aib, Ac5c; where aa1-aa 26 any two of which are optionally cyclized via their side chains to form a lactam bond; However, aa 16 , aa 17 , aa 18 , aa 22 , aa 23 , aa 24 or aa 25 is a linker amino acid U covalently attached to X.

[0229] In some embodiments of Formula 2-III, U is any linker amino acid described herein. In some embodiments, the compound of Formula 2-III is aa 12 and aa 16 are cyclized through their side chains to form a lactam bond. In some embodiments, the compound of formula 2-III is a compound 16 and aa 20 are compounds that are cyclized via their side chains to form lactam bonds.

[0230] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg11-Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-dodecyl beta-D-glucuronyl) 18 -NH2 (SEQ ID No. 180).

[0231] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-alkyl beta-D-glucuronyl) 18 -Aib 19 -NH2, where alkyl is dodecyl, tetradecyl, hexadecyl, or octadecyl (SEQ ID No. 283).

[0232] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-alkyl beta-D-glucuronyl) 18 -Ac4c 19-NH2, where alkyl is dodecyl, tetradecyl, hexadecyl, or octadecyl (SEQ ID No. 284).

[0233] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln * 16 -Aib 17 -Lys(N-epsilon-1'-alkyl beta-D-glucuronyl) 18 -Aib 19 -Lys * 20 -NH2, where Glu * 16 and Lys * 20 are attached through their side chains by lactams, and alkyl is dodecyl, tetradecyl, hexadecyl, or octadecyl (SEQ ID No. 285).

[0234] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Glu * 12 -Arg 13 -Trp 14 -Ile 15 -Lys * 16 -Aib 17 -Lys(N-epsilon-1'-alkyl beta-D-glucuronyl) 18 -Aib 19 -NH2, where Glu * 12 and Lys *16 are attached through their side chains by lactams, and alkyl is dodecyl, tetradecyl, hexadecyl, or octadecyl (SEQ ID No. 286).

[0235] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Phe 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-alkyl beta-D-glucuronyl) 18 -Aib 19 -NH2, where alkyl is dodecyl, tetradecyl, hexadecyl, or octadecyl (SEQ ID No. 287).

[0236] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-alkyl beta-D-glucuronyl) 18 -Aib 19 -Aib 20 -NH2, where alkyl is dodecyl, tetradecyl, hexadecyl, or octadecyl (SEQ ID No. 288).

[0237] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Glu * 12 -Arg 13 -Trp 14 -Ile 15 -Lys * 16 -Aib 17 -Lys(N-epsilon-1'-alkyl beta-D-glucuronyl) 18 -Aib 19 -Aib 20 -NH2, where Glu * 12 and Lys * 16 are attached through their side chains by lactams, and alkyl is dodecyl, tetradecyl, hexadecyl, or octadecyl (SEQ ID No. 289).

[0238] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-dodecyl beta-D-glucuronyl) 18 -Aib 19 -NH2 (SEQ ID No. 207).

[0239] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14-Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-tetradecyl beta-D-glucuronyl) 18 -Aib 19 -NH2 (SEQ ID No. 260).

[0240] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-hexadecyl beta-D-glucuronyl) 18 -Aib 19 -NH2 (SEQ ID No. 261).

[0241] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-octadecyl beta-D-glucuronyl) 18 -Aib 19 -NH2 (SEQ ID No. 262).

[0242] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11-Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-dodecyl beta-D-glucuronyl) 18 -Aib 19 -Aib 20 -NH2 (SEQ ID No. 275).

[0243] In some embodiments, the peptide product has the following structure: Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-1'-hexadecyl beta-D-glucuronyl) 18 -Aib 19 -Aib 20 -NH2 (SEQ ID No. 277).

[0244] In some embodiments, the peptide product is an active peptide product that binds to the PTH receptor (PTHR1).

[0245] In a specific embodiment, the peptide product of formula 2-IA described above and herein has the following structure:

[0246] [ka]

[0247] In the formula, R 1a is C1-C as described in Table 2 of Figures 2-1 to 2-4. 20 R' is a peptide as described in Table 2 of Figures 2-1 to 2-4, W of formula IA 2is -O-, and W in formula 2-IA 1 is —(C═O)NH— and is part of an amide bond to the peptide R′. In some such embodiments, R 1a is C6-C 20 In some such embodiments, R 1a is C8-C 20 In some such embodiments, R 1a is C8-C 20 In some such embodiments, R 1a is C8-C 18 In some such embodiments, R 1a is C8-C 16 It is an alkyl chain.

[0248] In the embodiments described above, the amino acid and / or amino moiety of peptide R′ (e.g., the amino group of an amino acid residue such as lysine, or a lysine within peptide R′) is used to form a covalent bond with a compound of the following structure:

[0249] [ka]

[0250] In the formula, R 1a is C1-C as described in Table 2 of Figures 2-1 to 2-4. 20 It is an alkyl chain.

[0251] In such cases, the amino acid having an amino moiety (e.g., a lysine residue in peptide R') used to form a covalent bond to compound A described above is the linker amino acid U attached to surfactant X having the structure of Formula 2-A. Thus, as an example, Lys(C12) in Table 2 of Figures 2-1 through 2-4 has the following structure:

[0252] [ka]

[0253] Also contemplated within the scope of the embodiments presented herein are peptide products of formula 2-IA derived from malturonic acid-based surfactants via attachment at either or both of the carboxylic acid functional groups. Thus, as an example, the peptide in Table 2 of Figures 2-1 to 2-4 includes a lysine linker amino acid attached to a malturonic acid-based surfactant, X, and having the following structure:

[0254] [ka]

[0255] It will be understood that in one embodiment, compounds of formula 2-IA are prepared by attaching lysine to group X, followed by attaching additional amino acid residues and / or peptides to the lysine-X compound to obtain compounds of formula 2-IA. It will be understood that other natural or unnatural amino acids described herein are also suitable for attachment to surfactant X, and for attaching additional amino acids / peptides to obtain compounds of formula 2-IA. It will be understood that in another embodiment, compounds of formula 2-IA are prepared by attaching a full-length or partial-length peptide to group X, followed by attaching additional amino acid residues and / or peptides to obtain compounds of formula 2-IA.

[0256] In specific embodiments, provided herein is a compound selected from the compounds of Table 2 in Figures 2-1 through 2-4.

[0257] Also provided herein is a pharmaceutical composition comprising a therapeutically effective amount of the peptide product described above, or an acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0258] 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 hydrofluoroalkane-like solvent containing submicron anhydrous α-lactose or other excipient.

[0259] Within the embodiments presented herein, reaction of amino acids and / or peptides comprising a linker amino acid U carrying a nucleophile with a group X comprising a leaving group or a functional group that can be activated to contain a leaving group, e.g., a carboxylic acid, or any other reactive group, is contemplated, thereby allowing covalent attachment of the amino acid and / or peptide to the surfactant X via the linker amino acid U to provide a peptide product of formula 2-IA.

[0260] Also contemplated within the embodiments presented herein is the reaction of amino acids and / or peptides containing a linker amino acid U bearing a leaving group or a functional group that can be activated to contain a leaving group, e.g., a carboxylic acid, or any other reactive group, with a group X containing a nucleophilic group, thereby allowing covalent attachment of the amino acid and / or peptide to surfactant X via the linker amino acid U to provide a peptide product of formula 2-IA.

[0261] It will be appreciated that in one embodiment, compounds of formula 2-IA are prepared by reaction of a linker amino acid U with X, followed by addition of additional residues to U to give a peptide product of formula 2-IA. In an alternative embodiment, compounds of formula 2-IA are prepared by reaction of a suitable peptide comprising a linker amino acid U with X, followed by optional addition of additional residues to U to give a peptide product of formula 2-IA.

[0262] Provided herein are methods for treating hypoparathyroidism, comprising administering to a subject in need thereof a therapeutically effective amount of a peptide product described above. In some embodiments, the hypoparathyroidism is associated with bone loss.

[0263] Also provided herein is a method for stimulating bone repair or supporting the survival of a bone implant, comprising administering to a subject in need thereof a therapeutically effective amount of the peptide product described above.

[0264] Also provided herein are covalently modified PTH or PTHrP peptides or analogs thereof comprising a hydrophilic group as described herein; and a hydrophobic group covalently attached to the hydrophilic group. In specific embodiments, the covalently modified peptide and / or protein product comprises a hydrophilic group that is a saccharide and a hydrophobic group that is a C1-C20 alkyl or aralkyl chain.

[0265] In one embodiment, a method is provided for chemically modifying a molecule by covalent attachment to a surfactant to increase or preserve the biological activity (e.g., receptor binding or enzymatic activity) of the composition or molecule. In some embodiments, the molecule is a peptide. The method can further include further modification, including covalently attaching the molecule in the composition to a polymer, such as polyethylene glycol.

[0266] In another embodiment, a method is provided for reducing or eliminating the immunogenicity of peptide and / or protein drugs by covalently attaching the peptide chain to at least one alkyl glycoside, wherein the alkyl has 1 to 30 carbon atoms.

[0267] Also provided are methods for treating osteoporosis, osteopenia, postmenopausal osteoporosis, Paget's disease, glucocorticoid-induced osteoporosis, senile osteoporosis, humoral hypercalcemia, and the like, which methods comprise administering to a vertebrate a pharmaceutical composition comprising a peptide covalently attached to at least one alkyl glycoside, wherein the alkyl has 1 to 30 carbon atoms, or even in the range of 6 to 16 carbon atoms, and wherein the covalent attachment of the alkyl glycoside to the peptide increases the stability, bioavailability, and / or duration of action of the drug.

[0268] In some embodiments, the covalently modified peptides and / or proteins are covalently modified PTH or PTHrP, or analogs thereof, modified to improve their pharmaceutical and medicinal properties by covalent modification with alkylglycoside surfactant moieties. These surfactant-modified analogs have increased steric hindrance that prevents proteolysis, slows absorption, and slows clearance from the body.

[0269] Some studies have shown that approximately 50 percent of patients with osteoporosis discontinue oral bisphosphonate therapy within one year. Many patients who discontinue these treatments do so due to side effects, including intolerance. Truncated recombinant parathyroid hormone 1-34 (rhPTH1-34) is commercially available as a bone anabolic agent, such as teriparatide (Lilly) (Brixen, KT, et al. (2004) Basic Clin Pharmacol Toxicol 94: 260-270; Dobnig, H. (2004) Expert Opin Pharmacother 5: 1153-1162), but poor compliance is a major problem. More recently, an antibody against a ligand that regulates osteoclast function (denosumab, Amgen) has been approved, but it has known serious side effects (serious skin infections, observed cases of osteonecrosis of the jaw, and severe inhibition of bone remodeling), some of which may have unclear long-term effects.

[0270] <Bone structure> Human bone architecture is maintained or elaborated by the coordinated functions of osteoclasts, which cause bone resorption, and osteoblasts, which lay down new bone matrix. Bone is the body's primary reservoir for calcium (a critical signaling ion), and a decrease in ambient extracellular calcium levels triggers increased PTH secretion via the calcium-sensing receptor on the parathyroid membrane. PTH binds to its receptor (PTHR1) present on the osteoblast membrane, leading to the expression of the receptor activator of nuclear factor-κB ligand (RANKL). RANKL binds to its receptor, RANK, on ​​osteoclast precursor cells, stimulating their differentiation and proliferation (Boyce, BF and Xing, L. (2007) Arthritis Res Ther 9 Suppl 1: S1). This leads to bone resorption (calcium mobilization from bone). PTH also acts to increase renal tubular calcium reabsorption and indirectly enhance intestinal calcium absorption via its stimulatory action on renal 1-alpha cholecalciferol hydroxylase (which increases circulating calcitriol), both of which act to produce a longer-term increase in circulating calcium ions.

[0271] The native form of human parathyroid hormone (hPTH) is an 84-amino acid peptide that plays a key role in maintaining calcium homeostasis in mammals (Rosen, CJ and Bilezikian, JP (2001) J Clin Endocrinol Metab 86: 957-964). A structurally related but independent hormone, parathyroid hormone-related protein (PTHrP), plays a paracrine role focused on bone growth in local tissues. Both hormones bind to the same receptor on osteoblasts, PTHR1, and result in the activation of multiple signaling pathways, including those regulated by increased cAMP levels.

[0272] However, the intermittent presence of PTH(1-34) only leads to osteoblast stimulation, lack of RANKL expression, and increased bone density. PTH(1-34) exhibits potent anabolic effects on the skeleton when given exogenously by intermittent injection. A small group of patients received teriparatide by daily subcutaneous injection for 6-24 months (Reeve, J., et al. (1980) Br Med J 280: 1340-1344), and paired bone biopsies revealed a large increase in the amount of cancellous bone in the intestine, with evidence of new bone formation and a suggestion that there was a dissociation between the rates of bone formation and resorption. Numerous studies have confirmed the development of bone tissue after daily injections of PTH analogs (Hodsman, AB, et al. (2005) Endocr Rev 26: 688-703; Cheng, Z., et al. (2009) J Bone Miner Res 24: 209-220). A review of the literature supports the observation that architectural improvements occur in the skeleton after daily teriparatide injections, in contrast to the skeletal architecture observed after treatment with antiresorptive agents, which act primarily by inhibiting osteoblast activity to reduce bone turnover, and therefore preserve rather than build new bone.

[0273] Continuous administration of exogenous PTH(1-34) rather than intermittent administration results in bone resorption. Thus, treatment with PTH(1-34) infusion for less than 6 hours results in increased bone density, while infusion for 8 hours or more results in bone resorption (Frolik, CA, et al. (2003) Bone 33: 372-379). Chronic administration of PTH(1-34) induces the expression of RANKL (activating RANK) on osteoblast precursor cells, thus stimulating their differentiation and proliferation (Boyce, BF and Xing, L. (2007) Arthritis Res Ther 9 Suppl 1: S1). This observation led to the current treatment paradigm: once-daily administration of PTH(1-34) by subcutaneous injection. More recently, a study using a daily PTH(1-34) infusion followed by a one-week withdrawal showed a significant increase in bone mineral density (Etoh, M. and Yamaguchi, A. (2010) J Bone Miner Metab, 28: 641-9). Teriparatide has a Cmax of 10 minutes and a half-life of 19 minutes (Frolik, CA, et al. (2003) Bone 33: 372-379). A more effective PTH analog would be expected to result in a more substantial increase in bone mineral density.

[0274] During toxicology studies with PTH(1-34) and PTH(1-84), a significant proportion of rats were observed to develop osteosarcomas, which began at approximately 20 months (Tashjian, AH, Jr. and Goltzman, D. (2008) J Bone Miner Res 23: 803-811). No treatment-related sarcomas have been reported in human trials with recombinant PTH(1-34), teriparatide (Forteo®). However, current treatment is limited to continuous daily subcutaneous injections for <2 years.

[0275] <PTHおよびPTHrP> In some embodiments, the methods and compositions described herein involve the use of PTH and / or PTHrP peptides and / or proteins and / or their analogs. All of the biological activity of intact human PTH (hPTH1-84) resides in the N-terminal sequence; most clinical studies have used the 34-amino acid peptide hPTH(1-34), known as teriparatide. The first two amino acids are essential for biological activity, and the anabolic properties of bone appear to be well maintained by the shortened fragment hPTH(1-31) or its cyclized lactam (Whitfield, JF and Morley, P. (1995) Trends Pharmacol Sci 16: 382-386). More recently, studies of sequences as short as hPTH(1-11) have shown activity and their EC 50 can be further modified to reduce the ATP content to the low nM range (Shimizu, M., et al. (2000) J Biol Chem 275: 21836-21843; Shimizu, N., et al. (2004) J Bone Miner Res 19: 2078-2086).

[0276] Studies of the interaction of PTH and PTHrP with PTH1R suggest that each ligand has two binding domains, one in the N-terminal 1-14 region and one in the C-terminal 15-34 region. The 1-14 portion has a more locally ordered structure and interacts with the 7-transmembrane domain of the receptor, while the 15-34 region is alpha-helical and interacts with the extracellular, N-terminal extension of the receptor. The N-terminal region of these peptides appears to be primarily responsible for receptor activity through interactions with this membrane-proximal region, whereas the C-terminal helical region has important binding interactions (Figures 2-1 to 2-4) with the extracellular domain of the receptor (Dean, T., et al. (2006) J Biol Chem 281: 32485-32495; Potetinova, Z., et al. (2006) Biochemistry 45: 11113-11121), resulting in higher ligand potency (Gardella, TJ, et al. (1994) Endocrinology 135: 1186-1194; Luck, MD, et al. (1999) Mol Endocrinol 13: 670-680). This leads to a two-domain model of binding that has been extended to other members of the family of class B GPCRs (Holtmann, M. H., et al. (1995) J Biol Chem 270: 14394-14398; Bergwitz, C., et al. (1996) J Biol Chem 271: 26469-26472; Runge, S., et al. (2003) J Biol Chem 278: 28005-28010).

[0277] Therefore, the present invention provides a covalently modified peptide or peptide analogue with a sequence that allows binding to PTH1R. The covalent modification of ligands in the PTH / PTHrP class relies on the development of an N-terminal ligand sequence that interacts with the membrane-proximal portion of PTHR1, linked to a C-terminal region that is truncated or simplified by using a surfactant moiety (e.g., 1-alkyl-glucuronic acid moiety, such as glucose-derived 1-alkylglucuronic acid). Because most of the interactions of the C-terminal region involve general hydrophobic interactions with the hydrophobic channel in the extracellular region of the receptor (Pioszak, AA and Xu, HE (2008) Proc Natl Acad Sci USA 105: 5034-5039; Pioszak, AA, et al. (2009) J Biol Chem 284: 28382-28391), the covalent modification described herein allows for increased binding interaction through low specificity interactions.

[0278] An example of improved cell stimulation by the surfactant-modified peptides described herein is illustrated in Figure 4. Accordingly, a substantially greater cAMP output (125%; superagonistic stimulation) is exhibited by cells stimulated with a dose of EU-232 (Figure 5) than by the internal standard, human PTHrP. Similarly, the coded sample of human PTHrP (Figure 6; EU-285) achieves only 100% of the maximal stimulation of the internal assay standard, human PTHrP. EU-232 is modified in the C-terminal region with a 1-dodecyl β-D-glucuronic acid moiety. Importantly, shorter peptide chains, or peptide chains of this size not modified with a 1-dodecyl β-D-glucuronic acid moiety, can be expected to exhibit reduced potency.

[0279] Similarly, in vivo responses to this surfactant-modified analog, EU-232, demonstrate high potency and extended duration of action (Figure 7). Blood phosphate levels were tested at various time points after dosing rats with saline (G1), 80 micrograms per kg of PTH (G2), 80 micrograms per kg of EU-232 (G3), or 320 micrograms per kg of EU-232 (G4). EU-232 demonstrates an extended duration of action, with the greatest statistically significant effect seen at the last time point in the assay (5 hours after dosing).

[0280] As shown in Figure 7, blood calcium levels were tested at various time points after dosing rats with saline (G1), 80 micrograms per kg of PTH (G2), 80 micrograms per kg of EU-232 (G3), or 320 micrograms per kg of EU-232 (G4). None of the groups were statistically significantly different from the control (G1). Importantly, the maximally effective dose and time point for EU-232 (G4; at 5 hours) showed no elevation, and therefore no trend toward hypercalcemia at the maximally effective dose. Hypercalcemia is a significant side effect seen after administration of PTH 1-34 and potent analogs of PTH (see).

[0281] The improvements described above and in the drawings, relating to surfactant modification to yield the peptides described herein, have important implications for their use in medicine. Such molecules are suitable for use with once-daily or less frequent administration to provide enhanced biological results compared to treatment with short-acting natural hormones such as PTH (T1 / 2 30 minutes by subcutaneous injection) or PTHrP. Surfactant-modified peptides such as EU-232 may be expected to exhibit greater biological effects when administered by intranasal inhalation due to the known effects of surfactants on nasal bioavailability.

[0282] Thus, in some embodiments, the surfactant-modified peptide products described herein reduce the occurrence of proteolysis. In some embodiments, covalent modification of PTH and / or PTHrP and / or analogs thereof can reduce the cost of producing therapeutic agents, and the presence of a covalently attached surfactant moiety provides advantageous pharmaceutical properties. In some embodiments, surfactant-modified PTH and / or PTHrP described herein prolongs the PK and duration of action (PD) of the resulting ligand compared to other known peptide ligands lacking such covalent modifications (e.g., ligands of Dean, et al. (Dean, T., et al. (2006) J Biol Chem 281: 32485-32495)). Long-term, safe administration of surfactant-modified PTH and / or PTHrP and / or analogs thereof is also contemplated within the scope of the embodiments presented herein.

[0283] In some instances, the N-terminal binding region ends around residue 14, and the helical region encompasses residues 16 and onward. Thus, ligands optimized in the 1-14 region by 1-alkyl glucuronic acid modification in the 15 region and beyond have high potency (1-alkyl modification) and high specific binding (N-terminus). The use of α-helix-stabilizing substitutions in the C-terminal region (Kaul, R. and Balaram, P. (1999) Bioorg Med Chem 7: 105-117) leads to higher helical content and higher potency. Commonly used α-helix stabilizers are the class of 1,1-dialkyl amino acids, such as Ala, Aib, Ac4c, and Ac5c (see definition below). Minimization of the PTH structure led to shortened analogs, in which the incorporation of a hindered α-helical stabilizer led to significant increases in potency (Shimizu, M., et al. (2000) J Biol Chem 275: 21836-21843). For example, substitution of Aib at positions 1 and 3 of PTH1-14 and PTH1-11 analogs led to increased potency (Shimizu, N., et al. (2001) J Biol Chem 276: 49003-49012). Incorporation of a more hindered α-helical stabilizer at position 1 led to further increases in potency (Shimizu, N., et al. (2004) J Bone Miner Res 19: 2078-2086). Substitution of Aib at various positions in PTH1-34 analogs, particularly at positions 12 and 13, also led to improved efficacy (Peggion, E., et al. (2003) Biopolymers 68: 437-457). However, Aib at positions 1 and 3 of the simple PTH1-11 sequence was shown to be poorly tolerated (Barazza, A., et al. (2005) J Pept Res 65: 23-35).Thus, in some embodiments, the α-helical content of PTH and / or PTHrP is a determinant of the stability of the peptide product (Marx, UC, et al. (1995) J Biol Chem 270: 15194-15202; Schievano, E., et al. (2000) Biopolymers 54: 429-447).

[0284] In some embodiments, the use of long side chains, such as Nε-(1′-dodecylbeta-D-glucuronyl)-lysine, in the covalent peptide modifications described herein destabilizes α-helices. Therefore, also within the scope of the embodiments presented herein are modifications that include α-helical stabilizers. Thus, in some embodiments, surfactant-modified peptide products described herein include a helix stabilizer (e.g., immediately N- and / or C-terminal to the surfactant substitution). In some embodiments, the α-helical stabilizer is located at position 12 in the PTH and / or PTHrP chain. By way of example only, the following table lists specific surfactant-modified peptide products (EU-212 to EU-282) that include an α-helical stabilizer:

[0285] In one embodiment, the covalently modified peptides suitable for the methods described herein are truncated analogs of PTHrP and / or its related hormone PTH, including, but not limited to: hPTH(1-34): Ser1-Val2-Ser3-Glu4-Ile5-Gln6-Leu7-Met8-His9-Asn10-Leu11-Gly12-Lys13-His14-Leu15-Asn16-Ser17-Met18-Glu19-Arg20-Val21-Glu22-Trp23-Leu24-Arg25-Lys26-Lys27-Leu28-Gln29-Asp30-Val31-His32-Asn33-Phe34-OH; (SEQ ID NO. 290) or hPTHrP(1-34): Ala1-Val2-Ser3-Glu4-His5-Gln6-Leu7-Leu8-His9-Asp10-Lys11-Gly12-Lys13-Ser14-Leu15-Gln16-Asp17-Leu18-Arg19-Arg20-Arg21-Phe22-Phe23-Leu24-His25-His26-Leu27-Ile28-Ala29-Glu30-Ile31-His32-Thr33-Ala34-OH (SEQ ID NO. 291).

[0286] In some embodiments, a peptide product described herein has the structure of Formula 2V:

[0287] [ka]

[0288] During the ceremony: Z is OH or -NH-R 3 where R 3 is H or C1-C 12 alkyl; or a PEG chain of less than 10 Da; aa1 is Ala, Ser, Val, Pro, Aib, Ac5c, or Deg; aa2 is Val; aa3 is Ser, Ala, Aib, Ac4c, or Deg; aa4 is Glu; aa5 is His, or Ile; aa6 is Gln or Cit; aa7 is Leu or Phe; aa8 is Leu, Met, or Nle; aa9 is His; aa 10 is Asp, Asn, Gln, Glu, Cit, Ala, or Aib; aa 11 is Lys, Leu, Ile, Arg, or hArg; aa12 is Gln, Ala, Glu, Lys, Aib, or Ac5c; aa 13 is Lys, or Arg; aa 14 is Ser, His, Trp, Phe, Leu, Arg, Lys, Glu, NaI(2), or at position aa 18 cyclized to; aa 15 are Ile, Leu, Aib; aa 16 are Gln, Asn, Glu, Lys, Ser, Cit, Aib, Ac5c, U(X); aa 17 are Asp, Ser, Aib, Ac4c, Ac5c, U(X), Z; aa 18 is absent, Leu, Gln, Cit, Aib, Ac5c, Lys, Glu, or U(X), or at position aa 14 cyclized to; aa 19 is absent, Arg, Glu, Aib, Ac4c, Ac5c, or U(X); aa 20 is absent, Arg, Glu, Lys, Aib, Ac4c, Ac5c, or U(X); aa 21 is absent, Arg, Val, Aib, Ac5c, Deg, or U(X); aa 22 is absent, Phe, Glu, Aib, Ac5c, Lys, U(X), or at position aa 18 or aa 26 cyclized to; aa 23 is absent or is Leu, Phe, Trp, or U(X); aa 24 is absent, His, Arg, Leu, Aib, Ac5c, or U(X); aa 25 is absent, His, Lys, Arg, or U(X); aa 26is absent, His, Lys, Arg, Aib, Ac5c, or at position aa 22 cyclized to; aa 27 is absent, Leu, or Lys; aa 28 is absent, Ile, or Leu; aa 29 is absent, Ala, Gln, Cit, or Aib; aa 30 is absent, Glu, Asp, or Aib; aa 31 is absent, Ile, Val, Aib, Ac5C, or U(X); aa 32 is absent, His, Aib, Ac5C, or U(X); aa 33 is absent, Thr, Asn, Aib, Ac5C, or U(X); aa 34 is absent, Ala, Phe, Aib, Ac5C, or U(X); aa 35 is absent, Aib, Ac5C, or U(X); aa 36 is absent, Aib, Ac5C, or U(X); U is the binding amino acid; and X is attached to the side chain of U with a surfactant; where aa1-aa 36 any two of which are optionally cyclized via their side chains to form a lactam bond; However, aa1-aa 36 One, or at least one, of the

[0289] In some embodiments, the peptide products described herein comprise aa1-aa2 of Formula V as described above (SEQ ID NO. 292). 20In some embodiments, the peptide products described herein comprise aa1-aa2 of Formula V as described above (SEQ ID NO. 293). 19 Includes:

[0290] In a specific embodiment, the linking amino acid U is a diamino acid such as Lys or Orn, X is a modified surfactant from the 1-alkylglycoside class attached to U, and Z is OH, or -NH-R 3 ), where R 3 is H or C1-C 12 or a PEG chain of less than 10 Da.

[0291] In some embodiments, a peptide product described herein has the structure of Formula 2-VI:

[0292] [ka]

[0293] During the ceremony: Z is OH or -NH-R 3 and R 3 is H, C1-C 12 alkyl or PEG chains less than 10 Da; aa1 is Aib, Ac5c, Deg; aa3 is Aib, Ac4c, Deg; aa5 is His, Ile; aa6 is Gln, Kit; aa7 is Leu, Phe; aa8 is Leu, Nle; aa 10 are Asp, Asn, Gln, Glu, Cit, Ala, Aib; aa 11 is Arg, hArg; aa 12 are Gly, Ala, Glu, Lys, Aib, and Ac5c; aa 13are Lys, Arg; aa 14 are Ser, His, Trp, Phe, Leu, Arg, Lys, and NaI(2); aa 15 are Ile, Leu, Aib; aa16 is Gln, Asn, Glu, Lys, Ser, Cit, Aib, U(X); aa 17 are Asp, Ser, Aib, Ac4c, Ac5c, U(X); aa 18 is absent or is Leu, Gln, Cit, Aib, Ac5c, Lys, Glu, U(X); aa 19 is absent or is Arg, Glu, Aib, Ac4c, Ac5c, U(X); aa 20 is absent or is Arg, Glu, Lys, Aib, Ac4c, Ac5c, U(X); aa 21 is absent or is Arg, Val, Aib, Ac5C, Deg, U(X); aa 22 is absent or is Phe, Glu, Lys or U(X); aa 23 is absent or is Leu, Phe, Trp or U(X); aa 24 is absent or is Leu, His, Arg, or U(X); aa 25 is absent or is His, Lys, or U(X), and aa 26 is absent or Aib, Ac5c; U is the binding amino acid; X is a modified surfactant from the 1-alkyl glycoside class attached to U, where the 1-alkyl group is a substituted or unsubstituted C1-C 20 Alkyl or substituted or unsubstituted C1-C 20 It is aralkyl; However, aa1-aa 26 One, or at least one, of the

[0294] In some embodiments, a peptide product described herein has the structure of Formula 2-VII:

[0295] [ka]

[0296] During the ceremony: Z is OH or -H2; aa1 is Aib, Ac5c; aa 12 are Ala, Glu, Lys, Aib, Ac5c; aa 14 are Trp, Phe, and NaI(2); aa 16 are Gln, Asn, Glu, Lys, Cit, U(X); aa 17 are Asp, Ser, Aib, Ac4c, Ac5c, U(X); aa 18 is absent or is Leu, Gln, Aib, U(X); aa 19 is absent or Arg, Glu, Aib, Ac4c, Ac5c; aa 20 is absent or Arg, Glu, Lys, Aib, Ac4c, Ac5c; aa 21 is absent or Arg, Val, Aib, Ac5C, Deg; aa 22 is absent or is Phe, Glu, Lys or U(X); aa 23 is absent or is Leu, Phe, Trp or U(X); aa 24 is absent or is His, Arg, or U; aa25 is absent or is His, Lys, or U, and aa 26 is absent or Aib, Ac5c; U is the binding amino acid; X is a modified surfactant from the 1-alkyl glycoside class attached to U, where the 1-alkyl group is a substituted or unsubstituted C1-C 20 Alkyl or substituted or unsubstituted C1-C 20 It is aralkyl; However, aa1-aa 26 One, or at least one, of the

[0297] In some embodiments, the peptide product has the structure of Formula III:

[0298] [ka]

[0299] During the ceremony: Z is OH or -NH; aa1 is Aib, or Ac5c; aa 12 is Ala, Aib, Glu, Lys, or Ac5c; aa 14 is Trp, Phe, Lys, Glu, or NaI(2); aa 16 is Gln, Asn, Glu, Lys, Cit, or U(X); aa 17 is Asp, Ser, Aib, Ac4c, Ac5c, or U(X); aa 18 is absent or Leu, Gln, Aib, Lys, Glu, or U(X); aa 19 is absent or Arg, Glu, Aib, Ac4c, or Ac5c; aa 20is absent or Arg, Glu, Lys, Aib, Ac4c, Ac5c; aa 21 is absent or Arg, Val, Aib, Ac5C, or Deg; aa 22 is absent or is Phe, Glu, Lys, or U(X); aa 23 is absent or Leu, Phe, Trp, or U(X); aa 24 is absent or His, Arg, or U(X); aa 25 is absent or His, Lys, or U(X); and aa 26 is absent or Aib, Ac5c; where aa1-aa 26 any two of which are optionally cyclized via their side chains to form a lactam bond; However, aa 16 , aa 17 , aa 18 , aa 22 , aa 23 , aa 24 or aa 25 is a linker amino acid U covalently attached to X.

[0300] In specific embodiments of formula 2-III above, X has the structure:

[0301] [ka]

[0302] During the ceremony: R 1a is a substituted or unsubstituted C1-C 30 is an alkyl group; R 1b , R 1c , and R 1d is H; W1 is —(C═O)—NH—; W 2 is -O-; and R 2 is a single bond.

[0303] In some of the embodiments described above, R1a is C1-C 20 Alkyl groups, C1-C18 alkyl groups, C1-C 16 Alkyl groups, or C1-C 12 In some embodiments of Formula 2-III, U is any linker amino acid described herein.

[0304] Within the embodiments presented herein, peptide products of Formula 2-IA, Formula 2-III, Formula 2-V, Formula 2-VI, or Formula 2-VII are contemplated, wherein the peptide product comprises one or more surfactant groups (e.g., group X having the structure of Formula I). ​​In one embodiment, the peptide product of Formula 2-IA, Formula 2-III, Formula 2-V, Formula 2-VI, or Formula 2-VII comprises one surfactant group. In another embodiment, the peptide product of Formula 2-IA, Formula 2-III, Formula 2-V, Formula 2-VI, or Formula 2-VII comprises two surfactant groups. In yet another embodiment, the peptide product of Formula 2-IA, Formula 2-III, Formula 2-V, Formula 2-VI, or Formula 2-VII comprises three surfactant groups.

[0305] Table 2 in Figures 2-1 through 2-4 illustrates specific examples of peptides suitable for covalent attachment to surfactants as described herein.

[0306] The present specification recognizes the importance of certain portions of SEQ ID NO. 170 for the treatment of diseases related to bone loss and / or hyperparathyroidism, including, but not limited to, osteoporosis, osteopenia, postmenopausal osteoporosis, Paget's disease, glucocorticoid-induced osteoporosis, inflammatory bone loss, implant settlement, osteonecrosis of the jaw, stem cell proliferation, senile osteoporosis, humoral hypercalcemia, and the like.

[0307] Thus, the amino acid residues aa1-aa of SEQ ID NO. 170 are referred to herein as 17 A method of treating a disease associated with bone loss (e.g., osteoporosis) and / or hyperparathyroidism in an individual in need thereof is provided, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0308] In a further embodiment, the amino acid residues aa1-aa of SEQ ID NO. 170 are provided herein. 18 A method of treating a disease associated with bone loss (e.g., osteoporosis) and / or hyperparathyroidism in an individual in need thereof is provided, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0309] In another embodiment, the amino acid residues aa1-aa of SEQ ID NO. 170 are provided herein. 19 A method of treating a disease associated with bone loss (e.g., osteoporosis) and / or hyperparathyroidism in an individual in need thereof is provided, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0310] In another embodiment, the amino acid residues aa1-aa of SEQ ID NO. 170 are provided herein. 20 A method of treating a disease associated with bone loss (e.g., osteoporosis) and / or hyperparathyroidism in an individual in need thereof is provided, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0311] In a further embodiment, administration of the PTH analogs described above causes an increase in bone mineral density.

[0312] The present specification recognizes the importance of certain portions of SEQ ID NOs. 171, 173, 174, 645, or 646 for the treatment of diseases related to bone loss and / or hyperparathyroidism, including, but not limited to, osteoporosis, osteopenia, postmenopausal osteoporosis, Paget's disease, glucocorticoid-induced osteoporosis, inflammatory bone loss, implant settlement, osteonecrosis of the jaw, stem cell proliferation, senile osteoporosis, humoral hypercalcemia, and the like.

[0313] Thus, the amino acid residues aa1-aa2 of SEQ ID NO. 171, 173, 174, 290 or 291 are included herein. 17 A method of treating a disease associated with bone loss and / or hyperparathyroidism in an individual in need thereof is provided, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0314] In further embodiments, the amino acid residues aa1-aa2 of SEQ ID NO. 171, 173, 174, 290 or 291 are included herein. 18

[0013] Provided is a method for treating a disease associated with bone loss (e.g., osteoporosis) and / or hyperparathyroidism in an individual in need thereof, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0315] In another embodiment, the amino acid residues aa1-aa2 of SEQ ID NO. 171, 173, 174, 290 or 291 are 19

[0013] Provided is a method for treating a disease associated with bone loss (e.g., osteoporosis) and / or hyperparathyroidism in an individual in need thereof, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0316] In another embodiment, the amino acid residues aa1-aa2 of SEQ ID NO. 171, 173, 174, 290 or 291 are 20A method of treating a disease associated with bone loss (e.g., osteoporosis) and / or hyperparathyroidism in an individual in need thereof is provided, comprising administering to the individual in need thereof a therapeutically effective amount of a PTH analog comprising:

[0317] In a further embodiment, administration of the PTH analog described above causes an increase in bone mineral density.

[0318] In any of the embodiments described above, the PTH analog is modified with a surfactant X of formula 2-I:

[0319] [ka]

[0320] During the ceremony: R 1a is independently at each occurrence a single bond, H, a non-natural, substituted or unsubstituted C-C 30 an alkyl group, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a moiety containing a steroid nucleus; R 1b , R 1c , and R 1d is independently at each occurrence a single bond, H, a non-natural, substituted or unsubstituted C-C 30 an 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 in each occurrence a single bond to U, H, a substituted or unsubstituted C-C 30Alkyl groups, substituted or unsubstituted alkoxyaryl groups, or substituted or unsubstituted aralkyl groups, -NH2, -SH, C2-C4-alkene, C2-C4-alkyne, -NH(C=O)-CH2-Br, -(CH2) m -maleimide, or -N3; n is 1, 2 or 3; m is 1-10.

[0321] In a specific embodiment, the PTH analog is modified with a surfactant X having the following structure:

[0322] [ka]

[0323] During the ceremony: R 1a is a substituted or unsubstituted C1-C 30 is an alkyl group; R 1b , R 1c , and R 1 d is H; W 1 is —(C═O)—NH—; W 2 is -O-; and R 2 is a single bond.

[0324] In some of the embodiments described above, R 1a is C1-C 20 Alkyl groups, C8-C 20 Alkyl group, C 12 -C 18 Alkyl group or C 14 -C 18 It is an alkyl group.

[0325] Modifications at the amino or carboxyl termini can be optionally introduced into peptides (e.g., PTH or PTHrP) (Nestor, JJ, Jr. (2009) Current Medicinal Chemistry 16: 4399-4418).

[0326] For example, peptides can be truncated or acylated at the N-terminus to produce peptide analogs that exhibit low potency, partial agonist, and antagonist activity, as seen for 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 contents of which are incorporated herein by reference. For example, deletion of the first six residues of bPTH results in antagonist analogs (Mahaffey, JE, et al. (1979) J Biol Chem 254: 6496-6498; Goldman, ME, et al. (1988) Endocrinology 123: 2597-2599), and similar effects to the peptides described herein result in potent antagonist analogs. Other modifications to the N-terminus of the peptide, such as the deletion or incorporation of a D-amino acid such as D-Phe, can also yield potent, long-acting agonists or antagonists when substituted with modifications described herein, such as long-chain alkyl glycosides. Such agonists and antagonists are also commercially useful and are within the scope of the contemplated embodiments described herein.

[0327] Within the embodiments presented herein, N-terminal truncations of PTH (e.g., 7-34 residue analogs) or PTHrP are contemplated, thereby providing inverse agonists (Gardella, TJ, et al. (1996) Endocrinology 137: 3936-3941) or antagonists. In some embodiments, inverse agonists and / or antagonists of PTH and / or PTHrP are useful for treating "humoral hypercalcemia" associated with a wide range of tumors.

[0328] Also contemplated within the scope of the embodiments described herein are surfactants covalently attached to peptide analogs, where the native peptide has been modified by acetylation, acylation, pegylation, ADP-ribosylation, amidation, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, covalent cross-linking of cysteines, formation of pyroglutamic acid, formylation, gamma cleavage, or the like. -Modified by carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, protein processing, phosphorylation, prenylation, racemization, glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-ribosylation, transfer RNA-mediated addition of amino acids to proteins, such as selenoylation, sulfation, arginylation, and 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 663: 48-62).

[0329] Also contemplated within the scope of the embodiments described herein are branched or cyclic peptides, with or without branching. Cyclic, branched, and branched cyclic peptides result from post-translational natural processes and can also be made by suitable synthetic methods. In some embodiments, any of the peptide products described herein include the peptide analogs described above that are then covalently attached to an alkyl-glycoside surfactant moiety.

[0330] Also contemplated within the scope of the embodiments presented herein are peptide chains substituted at appropriate positions by modification of the analogs claimed herein, for example, by acylation on the linker amino acid at the ε-position of Lys with fatty acids such as octane, decanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, 3-phenylpropanoic acid, or with saturated or unsaturated alkyl chains (Zhang, L. and Bulaj, G. (2012) Curr Med Chem 19: 1602-1618). Non-limiting illustrative examples of such analogs are as follows:

[0331] Ac5c1―Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-dodecanoyl) 18 -Aib 19 -NH2 (SEQ ID NO. 294)

[0332] Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15-Gln 16 -Aib 17 -Lys(N-epsilon-dodecanoyl) 18 -NH2 (SEQ ID NO. 295)

[0333] Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-palmitoyl) 18 -Aib 19 -NH2 (SEQ ID NO. 296)

[0334] Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Phe 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-dodecanoyl) 18 -Aib 19 -NH2 (SEQ ID NO. 297)

[0335] Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-tetradecanoyl) 18 -Aib 19 -NH2 (SEQ ID NO. 298)

[0336] Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-tetradecanoyl) 18 -Aib 19 -Aib 20 -NH2 (SEQ ID NO. 299)

[0337] Ac5c1-Val2-Aib3-Glu4-Ile5-Gln6-Leu7-Nle8-His9-Gln 10 -hArg 11 -Ala 12 -Arg 13 -Trp 14 -Ile 15 -Gln 16 -Aib 17 -Lys(N-epsilon-(Nalpha-dodecanoyl-L-glutamyl)) 18 -Aib 19 -NH2 (SEQ ID NO. 300), etc.

[0338] In other embodiments, the peptide chain is substituted at appropriate positions by reaction of a spacer and hydrophobic moiety, such as a steroid nucleus (e.g., a cholesterol moiety), on a linker amino acid (e.g., the sulfhydryl group of cysteine). In some such embodiments, the modified peptide further comprises one or more PEG chains. Non-limiting examples of nanoencapsulation are as follows:

[0339] [ka]

[0340] <GLPペプチドおよびアナログ>

[0341] Also provided herein, in some embodiments, are reagents and intermediates for the synthesis of peptides and / or proteins (e.g., modified GLP-1, glucagon, analogs of glucagon or GLP-1, etc.) modified by surfactant attachment.

[0342] Provided herein, in some embodiments, is a peptide product comprising a peptide, a peptide comprising a linker amino acid U, and a surfactant X covalently attached to at least one other amino acid.

[0343] [ka]

[0344] wherein surfactant X is a group of formula I:

[0345] [ka]

[0346] where R 1a is, at each occurrence, a single bond, H, substituted or unsubstituted C1-C 30 present independently in the alkyl group, substituted or unsubstituted alkoxyalkyl group, or substituted or unsubstituted aralkyl group, R 1b , R 1c and R 1d is, at each occurrence, a single bond, H, substituted or unsubstituted C1-C 30 each independently occurring in an alkyl group, a substituted or unsubstituted alkoxyalkyl group, or a substituted or unsubstituted aralkyl group; W 1 is present independently at each occurrence in —CH—, —CH—O—, —(C═O), —(C═O)—O—, —(C═O)—NH—, —(C═S)—, —(C═S)—NH—, or —CHS—; W 2 is —O—, —CH2— or —S—, R2 represents, at each occurrence, a single bond, H, a substituted or unsubstituted C1-C30 alkyl group, a substituted or unsubstituted alkoxyalkyl group, or a substituted or unsubstituted aralkyl group, —NH2, —SH, C2-C4-alkene, C2-C4-alkyne, —NH(C=O)-CH2-Br, —(CH2) m -maleimide, or -N3, n is 1, 2 or 3; m is 1-10, The peptide is selected from Formula 3-II:

[0347] [ka]

[0348] where: Z is OH (or -NH-R 3 ), where R 3 is H or C1-C 12 a substituted or unsubstituted alkyl or PEG chain of less than 10 Da; aa1 is His, N-Ac-His, pGlu-His, or NR 3 -His, aa2 is Ser, Ala, Gly, Aib, Ac4c or Ac5c, aa3 is Gln or Cit, aa4 is Gly or D-Ala, aa5 is Thr or Ser, aa6 is Phe, Trp, F2Phe, Me2Phe, or Na12; aa7 is Thr or Ser, aa8 is Ser or Asp, aa9 is Asp or Glu, aa 10 is Tyr, Leu, Met, NaI2, Bip or Bip2EtMeO, aa 11 is Ser, Asn, or U, aa 12is Lys, Glu, Ser, Arg or U, aa 13 is absent or is Tyr, Gln, Cit or U, aa 14 is absent or is Leu, Met, Nle, or U, aa 15 is absent or is Asp, Glu or U, aa 16 is absent or is Ser, Gly, Glu, Aib, Ac5c, Lys, Arg or U, aa 17 is absent or is Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c or U, aa 18 is absent or is Arg, hArg, Ala, Aib, Ac4c, Ac5c or U, aa 19 is absent or is Ala, Val, Aib, Ac4c, Ac5c or U, aa 20 is absent or is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c or U, aa 21 is absent or is Asp, Glu, Leu, Aib, Ac4c Ac5c or U; aa 22 is absent or is Phe, Trp, NaI2, Aib, Ac4c, Ac5c or U, aa 23 is absent or is Val, Ile, Aib, Ac4c, Ac5c or U, aa 24 is absent or is Gln, Ala, Glu, Cit or U, aa 25 is absent or is Trp, NaI2, or U, aa 26 is absent, or is Leu, or is U, aa 27is absent or is Met, Val, Nle, Lys, or U, aa 28 is absent or is Asn, Lys or U, aa 29 is absent or is Thr, Glycine, Aib, Ac4c, Ac5c or U, aa 30 is absent or is Lys, Aib, Ac4c, Ac5c or U, aa 31 is absent or is arginine, Aib, Ac4c, Ac5c or U, aa 32 is absent or is asparagine, Aib, Ac4c, Ac5c or U, aa 33 is absent or is arginine, Aib, Ac4c, Ac5c or U, aa 34 is absent or is asparagine, Aib, Ac4c, Ac5c or U, aa 35 is absent or is asparagine, Aib, Ac4c, Ac5c or U, aa 36 is absent or is Ile, Aib, Ac4c, Ac5C or U, aa 36 is absent or is Ala, Aib, Ac4c, Ac5C, or U; aa 37 is absent or is U, U is a natural or unnatural amino acid containing a functional group used for covalent attachment to surfactant X; where aa1-aa 37 any two of which are optionally cyclized through their side chains to form a lactam bond; And aa 11 -aa 37 is a linker amino acid U covalently attached to X.

[0349] In some embodiments, n is 1. In some embodiments, n is 2. The first glycoside may also be an OR of the first glycoside and the second glycoside. 1b , OR 1c OR 1d Any one of W 2 It is attached to the second glycoside by a bond between In some embodiments, n is 3. The first glycoside may also be an OR of the first glycoside and the second glycoside. 1b , OR 1c OR 1d Any one of W 2 a second glycoside and a third glycoside attached to the second glycoside by a bond between the second glycoside and the third glycoside 1b , OR 1c OR 1d Any one of W 2 It is attached to a third glycoside by a bond between

[0350] In one embodiment, the compound of formula IA is one in which X has the structure of formula 3-I:

[0351] [ka]

[0352] where: R 1a is H, a protecting group, a substituted or unsubstituted C1-C 30 an alkyl group or a steroid nucleus-containing moiety; R 1b , R 1c and R 1d represents each occurrence, H, a protecting group or a substituted or unsubstituted C1-C 30 each independently present in the alkyl group; W 1 exists independently at each occurrence (—CH2)—, —CHO, —(C═O), —(C═O)—O, —(C═O)—NH, —(C═S)—, —(C═S)—NH, or —CHS—; W 2is—O—,—S—; R 2 is a single bond, C2-C4 alkene, C2-C4 alkyne, or —(CH2) m -maleimide; and m1 to m10.

[0353] In another embodiment, the compound of formula IA is one in which X has the structure above.

[0354] [ka]

[0355] Thus, in the embodiment described above, R 2 is a single bond.

[0356] For example, in a typical embodiment of the structure of X described above, W1 is —C(═O)NH— and R 2 is W 1 and an amino acid residue U in a peptide (e.g., the amino group in the side chain of a lysine residue in a peptide).

[0357] In further embodiments, the compound of formula IA is a compound having the structure:

[0358] [ka]

[0359] For example, in exemplary embodiments of the structure of X described above, W 1 is -CH2-. Also, R 2 is the alkyl-linked maleimide functional group of X. Also, R 2 is attached to the appropriate position of an amino acid residue U within the peptide (e.g., a thiol group in a cysteine ​​residue of the peptide forms a thioether with the maleimide on X).

[0360] In yet other embodiments, the compound of formula IA has the structure:

[0361] [ka]

[0362] where R 1a is H, a protecting group, a substituted or unsubstituted C1-C 30 an alkyl group or a steroid nucleus-containing moiety; R 1b , R 1c and R 1d is, at each occurrence, H, a protecting group or a substituted or unsubstituted C-C 30 each independently present in the alkyl group; W1 is -(C=O)-NH-; W 2 is—O—; R 2 is a single bond.

[0363] In additional embodiments, the compound of formula IA is one in which X has the structure:

[0364] [ka]

[0365] where R 1a is a substituted or unsubstituted C1-C6 alkyl; R 1b , R 1c and R 1d is H; W 1 is -(C=O)-NH-; W 2 is—O—; and R 2 is a single bond.

[0366] In some embodiments described hereinabove, R 1a is a substituted or unsubstituted C1-C30 It is an alkyl group.

[0367] In some embodiments described hereinabove, R 1a is a substituted or unsubstituted C6-C 20 It is an alkyl group.

[0368] Also contemplated herein is an alternative embodiment, wherein X in formula 3-IA has the structure:

[0369] [ka]

[0370] For example, in the exemplary embodiment of the structure of X described above, W 1 is -S- and R 2 is C1-C 30 is an alkyl group, and W 2 is S and R 1a is W 2 and an appropriate portion of an amino acid residue U within the peptide (e.g., a thiol group in a cysteine ​​residue of the peptide forms a thioether with X).

[0371] In another exemplary embodiment of the structure of X described above, W 1 is -O- and R 2 is C1-C 30 is an alkyl group, and W 2 is O and R 1a is W 2 and the appropriate portion of an amino acid residue U in the peptide (e.g., a hydroxyl group in a serine or threonine residue of the peptide forms an ether with X).

[0372] In some embodiments, U is used to covalently attach to X and is a dibasic natural or artificial amino acid, the natural or artificial amino acid containing a thiol, the artificial amino acid containing an —N group, the artificial amino acid containing an acetylene group, or the artificial amino acid is —NHC(═O)—CH—Br or —(CH)—(CH) m -maleimide, where m is 1-10.

[0373] 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 attached to the peptide through an amide bond.

[0374] In some embodiments of the peptide product, surfactant X is selected from the group consisting of 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-diglucuronic acid, 1-octadecyl beta-D-diglucuronic acid, 1-hexadecyl beta-D-diglucuronic acid, 1-tetradecyl beta-D-diglucuronic acid, 1-dodecyl beta-D-diglucuronic acid, 1-decyl beta-D-diglucuronic acid, 1-octyl beta-D-diglucuronic acid, These peptides consist of functionalized 1-eicosyl beta-D-glucose, 1-octadecyl beta-D-glucose, 1-hexadecyl beta-D-glucose, 1-tetradecyl beta-D-glucose, 1-dodecyl beta-D-glucose, 1-decyl beta-D-glucose, 1-octyl beta-D-glucose, 1-eicosyl beta-D-maltoside, 1-octadecyl beta-D-maltoside, 1-hexadecyl beta-D-maltoside, 1-dodecyl beta-D-maltoside, 1-decyl beta-D-maltoside, 1-octyl beta-D-maltoside, and the like, and the peptide products are prepared by the formation of a bond between the aforementioned groups and a group on the peptide (e.g., the aforementioned —COOH group and an amino group of the peptide).

[0375] In some embodiments of the peptide product, U is the terminal amino acid of the peptide. In some embodiments of the peptide product, U is an unscheduled amino acid of the peptide. In some embodiments of the peptide product, U is a naturally occurring D-amino acid or L-amino acid. In some embodiments of the peptide product, U is an artificial amino acid. In some embodiments of the peptide product, U is selected from Lys, Cys, Orn, or an artificial amino acid that contains a functional group used for covalent attachment to surfactant X.

[0376] In some embodiments of the peptide product, the functional group used for covalent attachment of the peptide to surfactant X is —NH, —SH, —OH, —N, haloacetyl —(CH) m -maleimide (where m is 1-10), or an acetylene group.

[0377] In some embodiments, the side chain functional groups of two different amino acid residues are linked to form a cyclic lactam. For example, in some embodiments, a Lys side chain forms a cyclic lactam with a Glu side chain. In some embodiments, such a lactam structure is inverted and formed from a glutamic acid and a Lys. Such lactam bonds, in some instances, are known to stabilize alpha helical structures in peptides (Condon, SM et al. (2002) Bioorg Med Chem 10, pp. 731-736; Murage, EN et al. (2008) Bioorg Med Syn 16: 10106-12; Murage, EN et al. (2010) J Med Chem 53, pp. 6412-6412). In some embodiments, cysteine ​​residues can be linked via di-Thr bond formation to achieve a similar form of conformational constraint and to aid in the formation of helical structures (Li, Y. et al. (2011) Peptide 32:1400-1407). In some embodiments, side chain functional groups of two different amino acid residues can be linked to form a heterocycle resulting from a "click reaction" between side chain azide and alkyne functional groups to achieve a similar form of conformational constraint and stabilize the helical conformation (Le Chevalier Isaad A. et al. (2009) J Peptide Sci 15:451-4).

[0378] In some embodiments, the peptide product comprising a covalently attached alkyl glycoside is a covalently modified glucagon or analog thereof, and in some such embodiments, the peptide product comprises a covalently attached 1-O-alkyl beta-D-glucuronic acid, and the peptide is an analog of glucagon.

[0379] In some embodiments, the peptide product comprising a covalently linked alkyl glycoside is a covalently modified GLP-1 or analog thereof. In some such embodiments, the peptide product comprises a covalently linked 1-O-alkyl beta-D-glucuronic acid, and the peptide is an analog of GLP-1.

[0380] In some embodiments, the peptide product of formula IA has formula III-A, i.e.

[0381] [ka]

[0382] where Z is OH or -NHR 3 where R 3 is H or C1-C 12 a substituted or unsubstituted alkyl, or a PEG chain of less than 10 Da; aa1 is His, N-Ac-His, pGlu-His, or NR 3 -His; aa2 is Ser, Ala, Gly, Aib, Ac4c, or Ac5c; aa3 is Gln, or Cit; aa4 is Gly, or D-Ala; aa5 is Thr or Ser; aa6 is Phe, Trp, F2Phe, Me2Phe, or Na12; aa7 is Thr or Ser; aa8 is Ser or Asp; aa9 is ASp, or Glu; aa 10 is Tyr, Leu, Met, NaI2, Bip, or Bip2EtMeO; aa 11 is Ser, Asn, or U; aa 12is Lys, Glu, Ser, Arg, or U(X); aa 13 is absent or is Tyr, Gln, Cit or U(X); aa 14 is absent or is Leu, Met, Nle, or U(X); aa 15 is absent or is Asp, Glu, or U(X); aa 16 is absent or is Ser, Gly, Glu, Aib, Ac5c, Lys, Arg, or U(X); aa 17 is absent or is Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c, or U(X); aa 18 is absent or is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X); aa 19 is absent or is Ala, Val, Aib, Ac4c, Ac5c or U(X); aa 20 is absent or is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U(X); aa 21 is absent or is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U(X); aa 22 is absent or is Phe, Trp, NaI2, Aib, Ac4c, Ac5c, or U(X); aa 23 is absent or is Val, Ile, Aib, Ac4c, Ac5c or U(X); aa 24 is absent or is Gln, Ala, Glu, Cit, or U(X); aa 25 is absent or is Trp, NaI2, or U(X); aa 26 does not exist, or is Leu, or is U(X); aa 27 is absent or is Met, Val, Nle, Lys or U(X); aa 28 is absent or is Asn, Lys or U(X); aa 29 is absent or is Thr, Gly, Aib, Ac4c, Ac5c, or U(X); where aa1-aa 29 any two of which are optionally cyclized through their side chains to form a lactam bond; And 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 is a natural or artificial amino acid U covalently attached to X.

[0383] In some embodiments, the peptide product of Formula IA has the structure of Formula 3-III-B:

[0384] [ka]

[0385] where Z is OH or -NHR 3 where R 3 is H or substituted or unsubstituted C1-C 12 is alkyl; or a PEG chain of less than 10 Da; aa2 is Ser, Ala, Gly, Aib, Ac4c, or Ac5c; aa3 is Gln, or Cit; aa6 is Phe, Trp, F2Phe, Me2Phe, MePhe, or Na12; aa 10 is Tyr, Leu, Met, NaI2, Bip, or Bip2EtMeO; aa 11 is Ser, Asn, or U(X); aa 12 is Lys, Glu, Ser, or U(X); aa 13 is absent or is Tyr, Gln, Cit or U(X); aa 14 is absent or is Leu, Met, Nle, or U(X); aa 15 is absent or is Asp, Glu, or U(X); aa 16 is absent or is Ser, Gly, Glu, Aib, Ac4c, Ac5c, Lys, R, or U(X); aa 17 is absent or is Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c, or U(X); aa 18 is absent or is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X); aa 19 is absent or is Ala, Val, Aib, Ac4c, Ac5c or U(X); aa 20 is absent or is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U(X); aa 21 is absent or is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U(X); aa 22 is absent or is Phe, Aib, Ac4c, Ac5c, or is U(X), aa 23 is absent or is Val, Ile, Aib, Ac4c, Ac5c or U(X); where aa1-aa 23 any two of which are optionally cyclized through their side chains to form a lactam bond; and aa 16 , aa 17 , aa 18 , aa 19 , aa 20 , aa 21 , aa 22 , aa 23 or aa 24 or at least one of X is a natural or artificial amino acid U covalently attached to X.

[0386] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, U is any linker amino acid described herein.

[0387] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, U is any linker amino acid described herein. In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa 12 is lysine. In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa 14 is leucine.

[0388] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa 12 is lysine. In some embodiments of Formula IA, III-A, III-B, or Formula V, aa 14 is leucine.

[0389] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa 18 is a lysine residue added to X.

[0390] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa 17 is a homo-arginine (hArg) residue.

[0391] In some embodiments of Formula IA, III-A, III-B, or Formula V, aa 17 is a glycine residue.

[0392] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa2 is an Aib or Ac4c residue.

[0393] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide comprises one or more Aib residues.

[0394] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide comprises one or more Aib residues at the C-terminus.

[0395] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 619):

[0396] [ka]

[0397] where aa2 is Aib or Ac4c; aa 17 is Arg, hArg, or Gln; aa 19 is Aib, Ac4c or Ac5c; And the alkyl is C8 versus a C20 straight alkyl chain.

[0398] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 620):

[0399] [ka]

[0400] where aa2 is Aib or Ac4c; aa 17 is arginine, hArg, or glutamine, aa19 and aa20, individually Aib, Ac4c, or Ac5c; Also, alkyl is C8 to C 20 It is a straight alkyl chain.

[0401] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 621):

[0402] [ka]

[0403] where aa2 is Aib or Ac4c; aa 16 is Aib or Ac4c; aa 17 is Arg, hArg, or Gln; aa 19 is Aib, Ac4c or Ac5c; Also, alkyl is C9 to C 20 It is a straight alkyl chain.

[0404] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa 16 and aa 20 is cyclized to form the lactam bond.

[0405] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure: (SEQ. ID. No. 622):

[0406] [ka]

[0407] where aa2 is Aib or Ac4c; aa 16 and aa 20 are each individually Lys or Glu and are cyclized by their side chains to form a lactam bond; aa 17 is Arg, hArg, or Gln; aa 27 is Met or Nle; Also, alkyl is C8-C 20 It is a straight alkyl chain.

[0408] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 623):

[0409] [ka]

[0410] where aa2 is Aib or Ac4c, and aa 29 is Thr, Aib, Ac4c or Ac5c, and the 1'-alkyl group is selected from dodecyl, tetradecyl, hexadecyl or octadecyl; and the side chains on the amino acids at positions 16 and 20 are cyclized to form side chain lactams.

[0411] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, aa 12and aa 16 is cyclized to form the lactam bond.

[0412] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 624):

[0413] [ka]

[0414] where aa2 is Aib or Ac4c; aa 12 and aa16 are each individually either Lys or Glu and are cyclized via their side chains to form a lactam bond; aa 17 is Arg, or hArg; aa 19 and aa 20 are individually either Aib, Ac4c or Ac5c; Also, alkyl is C8-C 20 It is a straight alkyl chain.

[0415] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ. ID. NO. 625):

[0416] [ka]

[0417] where aa 12 and aa 16 are cyclized by their side chains to form lactam bonds; aa 17 is Arg or hArg; and alkyl is C 12 , C 14 , C 16 or C18 It is a straight alkyl chain.

[0418] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 626):

[0419] [ka]

[0420] where: aa 12 and aa 16 are each individually either Lys or Glu, and aa 12 and aa 16 are cyclized by their side chains to form lactam bonds; aa 17 is Arg or hArg; aa 19 and aa 20 are individually either Aib, Ac4c or Ac5c; and the 1'-alkyl group is selected from dodecyl, tetradecyl, hexadecyl or octadecyl.

[0421] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the structure (SEQ.ID.NO. 627):

[0422] [ka]

[0423] wherein aa2 is Aib or Ac4c, and aa6 is Me2Phe, MePhe, or phenylalanine; and 19 is Aib, Ac4c or Ac5c.

[0424] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 628):

[0425] [ka]

[0426] wherein aa2 is Aib or Ac4c and aa6 is Me2Phe, MePhe or Phe; aa 17 is Arg or hArg, and aa 19 or aa 20 is Aib, Ac4c or Ac5c.

[0427] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the structure (SEQ.ID.NO. 629):

[0428] [ka]

[0429] where aa 23 is Aib, Ac4c or Ac5c, and the 1'-alkyl group is selected from dodecyl, tetradecyl, hexadecyl or octadecyl.

[0430] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ. ID. NO. 630):

[0431] [ka]

[0432] where aa2 is Aib or Ac4c: aa6 is Me2Phe, MePhe, or Phe; aa 12 and aa 16 are each individually either Lys or Glu; Also, aa 16 and aa 20 are cyclized through their side chains to form lactam bonds; aa 17 is Arg, hArg, or Gln; aa 18 is Aib or Ala; aa 23 is Aib, Ac4c or Ac5c, and the 1'-alkyl group is selected from dodecyl, tetradecyl, hexadecyl or octadecyl.

[0433] In some embodiments of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, the peptide product has the following structure (SEQ.ID.NO. 631):

[0434] [ka]

[0435] where aa2 is Aib or Ac4c: aa6 is phenylalanine; aa 12 and aa 16 are each individually either Lys or Glu; Also, aa 12 and aa 16 are cyclized through their side chains to form lactam bonds; aa 17 is Arg or hArg; aa 19 is Aib, Ac4c or Ac5c; aa 20is Aib, Ac4c or Ac5c, and the 1'-alkyl group is selected from dodecyl, tetradecyl, hexadecyl or octadecyl.

[0436] In some embodiments, for any compound of formula 3-IA, 3-III-A, 3-III-B, or 3-V, X comprises a dodecyl alkyl chain.

[0437] In some embodiments, the peptide product is a biologically active peptide product that binds to GLP1R and / or binds to GLCR.

[0438] In specific embodiments, the peptide product of Formula 3-IA, 3-III-A, or 3-III-B or Formula 3-V described herein above has the following structure:

[0439] [ka]

[0440] where R 1a is C1-C as described in Table 1 of Figures 1-1 to 1-7. 20 R' is an alkyl chain, and R' is a peptide as described in Table 3 of Figures 8-1 to 8-5 and Table 4 of Figures 9-1 to 9-7, W of formula IA 2 is —O—, and W in formula IA 1 is —(C═O)NH— and is part of the amide bond to the peptide R′. In some such embodiments, R 1a is C6-C 20 is an alkyl chain, In some such embodiments, R 1a is C8-C 20 is an alkyl chain, In some such embodiments, R 1a is C 12 -C 20 is an alkyl chain, In some such embodiments, R 1a is C 12 -C 16 It is an alkyl chain.

[0441] In the above-described embodiments, the amino moiety 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 of the following structure:

[0442] [ka]

[0443] where R 1a is a C1-C20 alkyl chain as described above and in Table 3 of Figures 8-1 to 8-5 and Table 4 of Figures 9-1 to 9-7.

[0444] In such cases, an amino acid residue having an amino moiety (e.g., lysine in peptide R') is a linker amino acid U used to form a covalent bond to compound A as described above, and is attached to surfactant X. Surfactant X has the structure of formula A. Thus, as an example, Lys(C12) in Table 3 of Figures 8-1 to 8-5 and Table 5 of Figures 9-1 to 9-7 has the following structure:

[0445] [ka]

[0446] Also contemplated within the scope of the embodiments presented herein are peptide products of formula 3-IA derived from maltouronic acid-based surfactants via linkage at either or both of the carboxylic acid functional groups. Thus, as an example, the peptides in Table 3 of Figures 8-1 through 8-5 and Table 4 of Figures 9-1 through 9-7 contain a lysine linker amino acid linked to a maltouronic acid-based surfactant, X, and have the following structure:

[0447] [ka]

[0448] In one embodiment, compounds of formula 3-IA are prepared by adding lysine to group X, followed by the addition of amino acid residues and / or peptides to the lysine-X compound to obtain compounds of formula 3-IA. It is also understood that other natural or artificial amino acids described herein are suitable for addition to surfactant X, and for the addition of additional amino acids / peptides to obtain compounds of formula 3-IA. In another embodiment, compounds of formula 3-IA are prepared by adding a full-length or partial-length peptide to group X, followed by the optional addition of additional amino acid residues and / or peptides to obtain compounds of formula 3-IA.

[0449] In certain embodiments, provided herein are compounds selected from the compounds of Table 3 in Figures 8-1 through 8-5 and the compounds of Table 4 in Figures 9-1 through 9-7.

[0450] Also provided herein is a pharmaceutical composition comprising a therapeutically effective amount of the peptide product described above, or an acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0451] 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 include submicron anhydrous α-lactose or other excipients.

[0452] Contemplated within the embodiments presented herein is the reaction of amino acids and / or peptides comprising a linker amino acid U bearing a nucleophile and a group X comprising a bearing leaving group or functional group, which group X can be activated to include a leaving group, e.g., a carboxylic acid or other reactive group, thereby allowing the covalent attachment of the amino acid and / or peptide to surfactant X via the linker amino acid U to provide a peptide product of formula 3-IA.

[0453] Also contemplated within the scope of the embodiments presented herein is the reaction of amino acids and / or peptides that contain a linker amino acid U that carries a leaving group or functional group that can be activated to include a leaving group, such as a carboxylic acid or other reactive group, and that contains a group X that includes a nucleophile, thereby allowing the covalent attachment of the amino acid and / or peptide to a surfactant X via the linker amino acid U to provide a peptide product of formula IA.

[0454] It will be appreciated that in one embodiment, compounds of formula 3-IA are prepared by reacting X with a linker amino acid, U, followed by the addition of a residue to U to yield a peptide product of formula 3-IA. In an alternative embodiment, it will be appreciated that compounds of formula 3-IA are prepared by reacting X with a suitable peptide comprising a linker amino acid, U, followed by the optional addition of a further residue to U to yield a peptide product of formula 3-IA.

[0455] Provided herein are methods for treating conditions related to insulin resistance comprising the administration of a compound of formula 3-IA, 3-III-A, 3-III-B, or formula 3-V.

[0456] Provided herein is a method for treating diabetes, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, wound healing, insulin resistance, hyperglycemia, hyperinsulinemia, metabolic syndrome, diabetic complications, high blood levels of free fatty acids or glycerol, hyperlipidemia, obesity, hypertriglyceridemia, atherosclerosis, acute cardiovascular syndrome, infarction, ischemia-reperfusion or hypertension, the method comprising administering to an individual in need thereof a therapeutically effective amount of a peptide product as described herein above.

[0457] Provided herein is a method for reducing weight gain or inducing weight loss, the method comprising administering to a subject in need thereof a therapeutically effective amount of a peptide product described herein above.

[0458] Provided herein are methods for treating a mammalian condition characterized by insulin resistance or metabolic syndrome linked to obesity, comprising administering to an individual in need thereof a weight loss-inducing or insulin-sensitizing amount of a peptide product described herein above.

[0459] In some embodiments, the condition being treated is metabolic syndrome (Syndrome X). In some embodiments, the condition being treated is diabetes. In some embodiments, the condition being treated is hyperlipidemia. In some embodiments, the condition being treated is hypertension. In some embodiments, the condition being treated is vascular disease, including atherosclerosis, or systemic inflammation characterized by elevated C-reactive protein.

[0460] In some embodiments of the method, the effective amount of peptide product for administration is from about 0.1 μg / kg / day to about 100.0 μg / kg / day, from 0.01 μg / kg / day to about 1 mg / kg / day, or from 0.1 μg / kg / day to about 50 mg / kg / day.

[0461] Provided herein are methods of treating metabolic syndrome, or a component disease thereof, comprising administering to a subject in need thereof a therapeutically effective amount of a peptide product as described above. In some embodiments, the metabolic syndrome condition has progressed to diabetes.

[0462] Also provided herein are covalently modified GLCR and / or GLP1R binding peptides or analogs thereof, comprising a hydrophilic group as described herein; and a hydrophobic group covalently attached to the hydrophilic group. In specific embodiments, the covalently modified peptide and / or protein product comprises a hydrophilic group that is a saccharide, and a C1-C 20 It contains a hydrophobic group that is an alkyl or aralkyl chain.

[0463] <Insulin resistance> Risks associated with prolonged hyperglycemia include increased risk of microvascular complications, sensory neuropathy, myocardial infarction, stroke, macrovascular mortality, and all-cause mortality. Type 2 diabetes is also implicated in the causation of obesity, an additional global epidemic. At least $232 billion was spent on the treatment and prevention of diabetes worldwide in 2007, with three-quarters of that amount spent in industrialized countries on the treatment and general care of long-term complications, including efforts to prevent microvascular and macrovascular complications. In 2007, the estimated indirect cost of diabetes to the U.S. economy (lost productivity, disability, and premature death due to diabetes) was $58 billion.

[0464] Obesity leads to insulin resistance, a reduced ability of the body's cells to respond to insulin stimulation through a reduced number of insulin receptors and reduced binding of these receptors to critical intracellular signaling systems. The obese state further leads to "metabolic syndrome," a constellation of diseases with significant health consequences (insulin resistance, hypertension, atherosclerosis, etc.). When insulin resistance is diagnosed early enough, type 2 diabetes can be prevented or delayed by lifestyle changes, targeted at reducing caloric intake and body fat, and through pharmacotherapy, to normalize glycemic control. Despite treatment guidelines recommending early and aggressive intervention, many patients fail to reach glycemic control targets. Many factors contribute to the failure to successfully manage type 2 diabetes, including psychosocial and economic impacts and shortcomings in the efficacy, convenience, and tolerability profiles of available antidiabetic drugs. The peptide and / or protein products described herein are designed to overcome these shortcomings.

[0465] <Incretin effect> The "incretin effect" is used to describe the phenomenon whereby a glucose load delivered orally generates a 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 in healthy individuals, up to 70% of the insulin secretory response to a meal may be derived from the incretin effect.

[0466] Normally, incretin peptides are secreted on demand in response to ingested nutrients and have a short plasma half-life due to degradation by the dipeptidyl peptidase-IV (DPP-4) enzyme. In people with type 2 diabetes, pancreatic responsiveness to GLP-1 is impaired, but insulin secretory responses can be restored with pharmacological doses of human GLP-1 (Kieffer, TJ et al. (1995) Endocrinology 136:3585-3596). Additionally, GLP-1 promotes beta cell neogenesis and storage (Aaboe, K. et al. (2008) Diabets Obes Metab 10:994-1003). GLP-1 has additional beneficial effects on cardiac function; 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 in humans and reduces appetite (Toft-Nielsen, MB et al. (1999) Diabets Care, 22:1137-1143).

[0467] Treatment of diabetic patients with metabolically stable, long-acting analogs of GLP-1, as described, for example, in SR (2010) Pharmacotherapy 30: 609-624, suffers from problems related to ease of use and side effects such as nausea, pancreatitis, and the risk of thyroid cancer. GLP-1 analogs provide glucose-dependent stimulation of insulin secretion, leading to a reduced risk of hypoglycemia. In addition, as described below, many current treatments for diabetes cause weight gain, while GLP-1 analogs induce satiety and mild weight loss. Thus, in some embodiments, provided herein are long-acting, low-dose GLP-1 analogs, thereby reducing the side effects associated with current treatments.

[0468] Many peptide gut 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 of the preproglucagon gene product (prohormone convertase; PC), such as glucagon, GLP-1, glucagon-like peptide-2 (GPL-2), glicentin, and oxyntomodulin (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, glicentin, and OXM are co-secreted by L cells in the intestine in response to feeding. Proproglucagon is alternatively processed to generate glucagon in the α-cells of the pancreatic islets (PC2). The structure of OXM is essentially glucagon with an eight residue C-terminal extension.

[0469] In addition to stimulating insulin biosynthesis and glucose-dependent insulin secretion, GLP-1 and its stable mimetics (e.g., Byetta) also induce modest weight loss in animal models (Mack, CM et al. (2006) Int J Obes (Lond) 30:1332-1340) and in patients with type 2 diabetes (DeFronzo, RA et al. (2005) Diabates Care, 28:1092-1100; Buse, JB et al. (2010) Diabates Care, 33:1255-1261). Glucagon infusion reduces food intake in humans (Geary, N., et al. (1992) Am J Physiol 262:R975-980), while continuous glucagon treatment of adipose tissue also 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 a wide range of effects on energy metabolism (Hoeppner, KM et al. (2010) Physiol Behav). Glucagon or analogs can be used in diagnostic modalities for temporary paralysis of the gastric tract. Thus, at least two of the products from PC processing of proglucagon protein are linked to satiety and metabolic effects.

[0470] In rodents, repeated intraperitoneal administration of OXM, a proglucagon-like tertiary product, has been associated with decreased white adipose tissue and weight loss compared with control animals (Dakin, CL et al. (2004) Endocrinology 145:2687-2695). OXM reduces food intake by 19.3% during intravenous administration to normal weight humans, an effect that persists for more than 12 hours after injection (Cohen, MA et al. (2003) J Clin Endocrinol Metab 88:4696-4701). Treatment of volunteers over a 4-week period resulted in a sustained satiety effect and weight loss, reflecting a reduction in body fat (Wynne, K. et al. (2005) Diabets 54:2390-2395).

[0471] OXM is structurally more homologous to GLP-1 and glucagon and activates both the glucagon receptor (GCGR) and the GLP-1 receptor (GLP1R), but with 10- to 100-fold less potency than the eponymous ligand. Additionally, studies of OXM interaction with GLP1R suggest that supplementation with beta-arrestin may have different effects compared to GLP-1 (Jorgensen, R. et al. (2007) J Pharmacol Exp Ther 322:148-154), thus acting as a "biased" ligand. The specific receptor for OXM has been pursued for several years but remains elusive. It is hypothesized to act through the GLP1R and GCGR pathways. Thus, provided herein are methods for surfactant modification of intestinal peptides that can induce satiety, weight loss, and alleviation of insulin resistance and / or delay in the progression of prediabetes.

[0472] <glp-1> Given the complex and interactive behavior of the proproglucagon protein products on satiety and metabolism described above, researchers from several groups have investigated structure-activity relationships on the GLP-1 and glucagon structures. Residues throughout the sequence have been shown to be amenable to substitution. For example, substitution with Ala is well tolerated in the N-terminal region of GLP-1, particularly at positions 2, 3, 5, 8, 11, and 12 (Adelhorst, K. et al. (1994) J Biol Chem 269:6275-6278).

[0473] It has been shown that grafting the C-terminal residue from GLP-1 to the N-terminus of glucagon can achieve a chimeric analog capable of binding to GLP1R and GLCR (Hjorth, SA et al. (1994) J Biol Sci 269:30121-30124). Residues at position 3 (an acidic glutamic acid in GLP1 or a neutral glutamine in glucagon or OXM) decrease the affinity of glucagon (Runge, S. et al. (2003) J Biol Chem 278:28005-28010) or OXM (Pocai, A. et al. (2009) Diabets 58:2258-2266) for GLP1R. The effects on the metabolic profile of animals treated with stable analogs of GLP-1 or glucagon or OXM with Gln at position 3 or OXM have been investigated (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) Diabets 58:2258-2266). These analogs were designed to have opposing effects on both GLP1R and GCGR (Day, JW et al., U.S. Patent Application Publication No. 2010 / 0190701A1).

[0474] Chimeric analogs should have the desired effects of the parent hormones acting on their receptors, and thus, clearly mimic the effects of OXM acting on both GLP-1R and GLCR: lipolysis and glucose-dependent insulin secretion and satiety, coupled with increased fat burning by glucagon. Analogs have been shown to induce the desired effects of weight loss and increased fat burning. Such properties are attractive for the treatment of obesity. However, a major challenge in obesity treatment is compliance. While currently known full-length analogs of glucagon and OXM can result in weight loss due to their affinity for both GLP-1R and GLCR, respectively, these analogs are not optimized for high bioavailability, pharmaceutical properties, and convenient delivery to patients required for optimal drug treatment regimens. Thus, provided herein are analogs of gut peptides (e.g., GLP, OXM, glucagon, etc.) that enable high bioavailability and / or long-term, durable effects for improved therapeutic outcomes in the treatment of conditions such as obesity and / or diabetes and / or metabolic syndrome.

[0475] Additional factors in optimizing the treatment of metabolic syndrome and diabetes with molecules like OXM relate to the duration of treatment and the amount of glucagon activity. For example, continuous treatment with analogs that activate GLP-1 and glucagon receptors (OXM pharmacological properties) can result in very large and rapid loss of fat mass (Day, JW et al. (2009) Nat Chem Biol 5:749-757), but it can also cause the loss of lean muscle mass (Kosinski, JR et al. (2012) Obesity (Silver Spring): doi:10.1038 / oby.2012.67). This is a disadvantage for this class of drugs. For example, in a study by Kosinski, JR et al., the naturally extracted hormone OXM, administered continuously for 14 days via an Alzet minipump, resulted in a 30% decrease in fat mass and a 7% decrease in lean muscle mass.

[0476] Glucagon action is known to stimulate glycogenolysis, lipolysis, and increased fat burning, but may also have catabolic effects on muscle. Successful treatment using agents that combine GLP-1 and glucagon action (OXM properties) requires optimal satiety and enhanced glucose-dependent insulin secretion from a GLP-1 analog with a judicious amount of glucagon action (fat burning). In addition, discontinuous use of such agents provides the desired clinical characteristics of moderate and continuous weight loss through loss of fat mass with minimal loss of lean mass. Provided herein are molecules with the desired combination of GLP-1 and OXM action, as well as tunable pharmacokinetic / pharmacological properties, allowing for optimal use in therapy (e.g., in metabolic syndrome, diabetes, obesity, etc.).

[0477] In one embodiment, the compounds of formulas 3-IA, 3-III-A, 3-III-B, and 3V are designed to provide either glucagon-like activity or GLP-1-like activity. The compounds of formulas 3-IA, 3-III-A, 3-III-B, and 3V provide tunable activity. For example, in one embodiment, the peptide products described herein have an EC50 of less than about 500 nM, preferably less than about 50 nM, and more preferably less than about 20 nM at both glucagon and GLP-1 receptors (e.g., the compounds in Table 3 of Figures 8-1 to 8-5 and Table 4 of Figures 9-1 to 9-7). In another example, the peptide products described herein (e.g., compounds in Table 3 of Figures 8-1-8-5 and Table 4 of Figures 9-1-9-7) are more potent at the GLP-1 receptor (e.g., less than 10 nM, preferably less than 10 nM, more preferably less than 1 nM) and less potent at the glucagon receptor (e.g., EC50 less than about 50 nM, preferably less than about 20 nM, more preferably less than 5 nM). This balance of biological activity allows for a judicious amount of glucagon action to be pooled, thereby allowing fat burning to occur while also retaining the beneficial effects of enhanced glucose-dependent insulin secretion. OXM is structurally more homologous to GLP-1 and glucagon and activates both the glucagon receptor (GCGR) and the GLP-1 receptor (GLP1R). Thus, in some embodiments, compounds of formulas 3-IA, 3-III-A, 3-III-B, and 3V provide tunable OXM-like biological activity. In certain embodiments, the peptide products described herein include peptides having amino acid residues 1-17 of GLP-1 and / or analogs thereof (e.g., modified artificial amino acid substitutions as described herein, cyclized lactam linkages as described herein, surfactant modifications as described herein, or combinations thereof).In certain other embodiments, the peptide products described herein include peptides having amino acid residues 1-16 of GLP-1 and / or analogs thereof (e.g., modified artificial amino acid substitutions as described herein, cyclized lactam linkages as described herein, surfactant modifications as described herein, or combinations thereof). In additional embodiments, the peptide products described herein include peptides having amino acid residues 1-186 of GLP-1 and / or analogs thereof (e.g., modified artificial amino acid substitutions as described herein, cyclized lactam linkages as described herein, surfactant modifications as described herein, or combinations thereof). Additionally, the peptide products described herein include one or more residues (e.g., Aib, Ac4C) that provide helical stabilization for the designed compounds of formulas 3-IA, 3-III-A, 3-III-B, and 3V, and the compounds in Table 3 of Figures 8-1 through 8-5 and Table 4 of Figures 9-1 through 9-7.

[0478] The glucagon subfamily of ligands is believed to bind to the receptor in a two-domain mode common to a number of class B receptors (secretin class, G protein-coupled receptors (GPCRs)). For GLP-1, there appears to be an N-terminal region, from residue 1 to approximately residue 16, that binds to the top of the transmembrane helices (juxtamembrane region), and a helical C-terminal region, from residues 17 to 31, that binds to the large, extracellular, N-terminal extension (ECD) of the receptor. Binding of these ligands highlights the fact that N-terminally truncated analogs of these peptide ligands can still retain the intrinsic binding ability and selectivity for just 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. It has recently been shown that short N-terminal analogs of GLP-1 can be both potent binders as well as receptor activators (Mapelli, C. et al. (2009) J Med Chem 52:7788-7799; Haque, TS et al. (2010) Peptide 31:950-955; Haque, TS et al. (2010) Peptide 31:1353-1360).

[0479] In addition, the X-ray crystal structure of the N-terminal region of GLP1R with a truncated antagonist of the GLP-1 mimetic analog exendin-4 (Byetta) bound in this region (Runge, S. et al. (2008) J Biol Chem 283:11340-7) indicates that the critical ligand-binding region in the ECD is highly hydrophobic (Figure 10). The sequence of exendin-4 beyond Glu15 is located in this very hydrophobic region (Val). 19* , Phe 22* , Trp 25* , Leu 26* ) interact as an amphipathic helix. In one embodiment, GLP-1 or glucagon is modified to bind to the GLCR by truncating the N-terminal fragment and covalently linked to a surfactant. The hydrophobic 1'-alkyl portion of the surfactant mimics and replaces the C-terminal region of the native hormone ligand, increasing peptide potency, efficacy, and duration of action. Additionally, such analogs have a major advantage due to their smaller size, which reduces their complexity, synthetic cost, and susceptibility to proteolysis. Additionally, smaller peptides are more easily absorbed by the nasal mucosa or enterocyte barrier.

[0480] Hypoglycemia is a potentially life-threatening condition of low blood sugar, and more aggressive treatment of high blood sugar through intensive insulin therapy is seen in more and more patients. Hypoglycemia occurs when blood sugar levels fall too low to provide the brain and muscles with enough energy for bodily activity. Glucagon can be used to treat this condition by stimulating the liver to break down glycogen, which produces glucose and raises blood sugar levels toward normal levels. Glucagon analogs that retain the ability to activate GLCR can be used to achieve the desired effect on blood sugar levels.

[0481] GLP-1 analogs that activate GLP1R stimulate the production and release of insulin from the pancreas in the presence of elevated blood glucose levels. This action results in effective control and normalization of blood glucose levels, as seen with current products such as exenatide (Byetta®). In addition, such products appear to produce appetite reduction and slow the movement of food from the stomach. Thus, they are effective in treating diabetes through multiple mechanisms. Analogs that combine the effects of glucagon and GLP-1, activating both GLCR and GLP1R, may provide benefits in the treatment of diabetes through their combined action to suppress appetite, release insulin in a glucose-dependent manner, help protect against hypoglycemia, and accelerate fat burning.

[0482] Such methods of treating hyperglycemia, including diabetes, diabetes mellitus type I, diabetes mellitus type II, or gestational diabetes, insulin-dependent or non-insulin-dependent, are expected to be useful in reducing diabetic complications, including nephropathy, retinopathy, and vascular disease. Applications in cardiovascular disease include microvascular as well as macrovascular disease (Davidson, M. H. (2011), J. Cardiol. 108 (suppl): 33B-41B; Gejl, M. et al. (2012) J. Clin. Endocrinol. Metab. 97 (doi:10.1210 / jc.2011-3456), and the treatment of myocardial infarction. Such methods of 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 reducing obesity-related complications, including vascular disease (coronary artery disease, stroke, peripheral vascular disease, ischemia-reperfusion, etc.), hypertension, the onset of type II diabetes, hyperlipidemia, and musculoskeletal disorders.

[0483] As used herein, the term glucagon or GLP-1 analogue includes all pharmaceutically acceptable salts or esters thereof.

[0484] In one embodiment, peptides that are covalently modified and suitable for the methods described herein are truncated analogs of glucagon and / or the related hormone GLP-1, including but not limited to:

[0485] [ka]

[0486] In some embodiments, the peptide products described herein have the structure of Formula 3-V:

[0487] [ka]

[0488] where U is the binding amino acid; X is surfactant-linked to the side chain of U; Z is OH or -NHR 3 where R 3 is H or C1-C 12 is a substituted or unsubstituted alkyl of the formula: aa1 is His, N-Ac-His, pGlu-His, or NR 3 -His; aa2 is Ser, Ala, Gly, Aib, Ac4c, or Ac5c; aa3 is Gln, or Cit; aa4 is Gly, or D-Ala; aa5 is Thr or Ser; aa6 is Phe, Trp, F2Phe, Me2Phe, or NaI(2); aa7 is Thr or Ser; aa8 is Ser or Asp; aa9 is Asp, or Glu; aa 10 is Tyr, Leu, Met, NaI(2), Bip, or Bip2EtMeO; aa 11 is Ser, Asn, or U(X); aa 12 is Lys, Glu, Ser, Arg, or U(X); aa 13 is absent, Tyr, Gln, Cit or U(X); aa 14 is absent, Leu, Met, Nle, or U(X); aa 15 is absent, Asp, Glu or U(X); aa 16 is absent, Ser, Gly, Glu, Aib, Ac5c, Lys, Arg, or U(X); aa 17 is absent, Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c, or U(X); aa 18 is absent, Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X); aa 19 is not present, Ala, Val, Aib, Ac4c, Ac5c or U(X); aa 20 is absent, Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U(X); aa 21 is absent, Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U(X); aa 22 is absent, Phe, Trp, Nal(2), Aib, Ac4c, Ac5c, or U(X); aa 23 does not exist, is Val, Ile, Aib, Ac4c, Ac5c, or U(X); aa 24 is absent, Gln, Ala, Glu, Cit, or U(X); aa 25 is absent, Trp, NaI(2) or U(X); aa 26 does not exist, Leu, U(X); aa 27 is absent, Met, Val, Nle, Lys, or U(X); aa 28 is absent, Asn, Lys or U(X); aa 29 is absent, Thr, Gly, Aib, Ac4c, Ac5c, or U(X); aa 30 is absent, Lys, Aib, Ac4c, Ac5c, or U(X); aa 31 is absent, Arg, Aib, Ac4c, Ac5c, or U(X); aa 32 does not exist, is Asn, Aib, Ac4c, Ac5c, or U(X); aa 33 is absent, Arg, Aib, Ac5c, or U(X); aa 34 does not exist, is Asn, Aib, Ac4c, Ac5c, or U(X); aa 35 does not exist, is Asn, Aib, Ac4c, Ac5c, or U(X); aa 36 does not exist, is Ile, Aib, Ac4c, Ac5C, or U(X); aa 36 is absent, Ala, Aib, Ac4c, Ac5C, or U(X); aa 37 does not exist or U(X); However, aa 11 or aa 37 We have stipulated that one, or at least one, of the following is U(X).

[0489] In a specific embodiment, the linking amino acid U is a diamino acid such as lymphocyte or ornithine, and X is a surfactant modified from the 1-alkylglycoside class linked to U, where Z is OH or -NHR. 3 where R 3 is H or C1-C 12 or a PEG chain of less than 10 Da.

[0490] In some embodiments, a peptide product described herein has the structure of Formula III-B:

[0491] [ka]

[0492] , where Z is OH (or -NHR 3 ), where R 3 is H or substituted or unsubstituted C1-C 12 is alkyl; or PEG chains less than 10 Da; aa2 is Ser, Ala, Gly, Aib, Ac4c, or Ac5c; aa3 is Gln, or Cit; aa6 is Phe, Trp, F2Phe, Me2Phe, MePhe, or Na12; aa 10 is Tyr, Leu, Met, NaI2, Bip, or Bip2EtMeO; aa 11 is Ser, Asn, or U; aa 12 is Lys, Glu, Ser, or U(X); aa 13 is absent or is Tyr, Gln, Cit or U(X); aa 14 is absent or is Leu, Met, Nle, or U(X); aa 15 is absent or is Asp, Glu, or U(X); aa 16 is absent or is Ser, Gly, Glu, Aib, Ac4c, Ac5c, Lys, R, or U(X); aa 17 is absent or is Arg, hArg, Gln, Glu, Cit, Aib, Ac4c, Ac5c, or U(X); aa 18 is absent or is Arg, hArg, Ala, Aib, Ac4c, Ac5c, or U(X); aa 19 is absent or is Ala, Val, Aib, Ac4c, Ac5c or U(X); aa 20 is absent or is Gln, Lys, Arg, Cit, Glu, Aib, Ac4c, Ac5c, or U(X); aa 21 is absent or is Asp, Glu, Leu, Aib, Ac4c, Ac5c, or U(X); aa 22 is absent, or Phe, Aib, Ac4c, Ac5c, or U(X)aa23 is absent, or is Val, Ile, Aib, Ac4c, Ac5c, or U(X); Here, aa1 to aa 23 any two of aa are optionally cyclized through their side chains to form a lactam bond; and 16 , aa 17 , aa 18 , aa 19 , aa 20 , aa 21 , aa 22 , aa 23 or aa 24 or at least one of X is a natural or artificial amino acid U covalently attached to X.

[0493] In some specific embodiments of Formula 3-III-A, Formula 3-III-B, and Formula 3-V, X has the structure:

[0494] [ka]

[0495] where: R 1a is a substituted or unsubstituted C1-C6 alkyl; and R 1b , R 1c and R 1d is H; W 1 is -(C=O)-NH-; W 2 is—O—; and R 2 is a single bond.

[0496] In some of the embodiments described above, R 1a is C1-C 20 Alkyl groups, C8-C 20 Alkyl group, C 12 -C 18 Alkyl group or C 14 -C 18 It is an alkyl group.

[0497] In some embodiments of Formula 3-III-B, U is any linker amino acid described herein. Table 3 in Figures 8-1 through 8-5 and Table 4 in Figures 9-1 through 9-7 illustrate specific examples of peptides covalently linked by surfactants as described herein.

[0498] Contemplated within the scope of the embodiments presented herein are peptide products of Formula 3-IA, Formula 3-III-A, Formula 3-III-B, or Formula 3-V, where the peptide product comprises one or more surfactant groups (e.g., group X having the structure of Formula 3-I). In one embodiment, the peptide product of Formula 3-IA, Formula 3-III-A, Formula 3-III-B, or Formula 3-V comprises one surfactant group. In another embodiment, the peptide product of Formula 3-IA, Formula 3-III-A, Formula 3-III-B, or Formula 3V comprises two surfactant groups. In yet another embodiment, the peptide product of Formula 3-IA, Formula 3-III-A, Formula 3-III-B, or Formula 3V comprises three surfactant groups.

[0499] The importance of certain portions of SEQ.ID.No. 632 for the treatment of conditions related to insulin resistance and / or cardiovascular conditions is recognized herein. Accordingly, provided herein is a method for administering amino acid residues aa1-aa2 of SEQ.ID.No. 632 to an individual in need thereof. 17 A method for treating diabetes in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0500] In a further embodiment, provided herein is a method of administering to an individual in need thereof a human ovarian tumor suppressor comprising ... 18 A method for treating diabetes in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0501] In another embodiment, provided herein is a method of administering amino acid residues aa1-aa2 of SEQ.ID.NO. 632 to an individual in need thereof. 19 A method for treating diabetes in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0502] In another embodiment, provided herein is a method for treating diabetes in an individual in need thereof, comprising administering to the individual in need thereof a therapeutically effective amount of a glucagon analog comprising amino acid residues aa1-aa20 of SEQ.ID.NO.632.

[0503] In a further embodiment, administration of the glucagon analog described above causes weight loss.

[0504] The importance of certain portions of SEQ ID NO. 303 for the treatment of conditions related to insulin resistance and / or cardiovascular conditions is recognized herein. Accordingly, provided herein is a method for administering amino acid residues aa1-aa2 of SEQ ID NO. 303 to an individual in need thereof. 17

[0013] Accordingly, provided herein is a method for treating diabetes in an individual in need thereof, comprising administering to an individual in need thereof a therapeutically effective amount of a glucagon analog comprising amino acid residues aa1-aa2 of SEQ.ID.NO. 303. 17

[0013] Accordingly, provided herein is a method for treating diabetes in an individual in need thereof, comprising administering to an individual in need thereof a therapeutically effective amount of a glucagon analog comprising amino acid residues aa1-aa2 of SEQ.ID.NO. 303. 17

[0013] Accordingly, provided herein is a method for treating diabetes in an individual in need thereof, comprising administering to an individual in need thereof a therapeutically effective amount of a glucagon analog comprising amino acid residues aa1-aa2 of SEQ.ID.NO. 303. 17 A method for treating diabetes in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0505] In a further embodiment, administration of the glucagon analog described above causes weight loss.

[0506] In any of the above-described embodiments, the glucagon analog is modified with a surfactant X of formula 3-I:

[0507] [ka]

[0508] where: R 1a is, at each occurrence, a single bond, H, substituted or unsubstituted C1-C 30 independently present in the alkyl group, substituted or unsubstituted alkoxyalkyl group, substituted or unsubstituted aralkyl group, or steroid nucleus-containing moiety; R 1b , R 1c and R 1d is, at each occurrence, a single bond, H, substituted or unsubstituted C1-C 30 each independently present in an alkyl group, a substituted or unsubstituted alkoxyalkyl group, or a substituted or unsubstituted aralkyl group; W1 is present independently at each occurrence in —CH2—, —CHO—, —(C═O), —(C═O)—O—, —(C═O)—NH—, —(C═S)—, —(C═S)—NH—, or —CHS—; W 2 is —O—, —CH2— or —S—; R 2 Each occurrence of represents a single bond to U, H, substituted or unsubstituted C1-C 30 Alkyl groups, substituted or unsubstituted alkoxyalkyl groups, or substituted or unsubstituted aralkyl groups, —NH2, —SH, C2-C4-alkene, C2-C4-alkyne, —NH(C=O)-CH2-Br, —(CH2) m -maleimide, or -independently present at N3; n is 1, 2 or 3; m is 1 to 10.

[0509] In certain embodiments, the glucagon analog is modified with a surfactant (X having the structure:

[0510] [ka]

[0511] where: R 1a is a substituted or unsubstituted C1-C6 alkyl; and R 1b , R 1c and R 1d is H; W 1 is(C=O)-NH―; W 2 is—O—; Also, R 2 is a single bond.

[0512] In some of the embodiments described above, R 1a is C1-C 20 Alkyl groups, C8-C 20 Alkyl group, C 12 -C 18 Alkyl group or C 14 -C 18 It is an alkyl group.

[0513] Also provided herein is a method for treating cardiovascular disease in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a glucagon analog comprising amino acid residues aa1-aa17 of SEQ. ID. NO. 632.

[0514] Also provided herein is a method for administering amino acid residues aa1-aa2 of SEQ. ID. NO. 632 to an individual in need thereof. 18 A method for treating cardiovascular disease in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0515] Also provided herein is a method for administering amino acid residues aa1-aa2 of SEQ. ID. NO. 632 to an individual in need thereof. 19 A method for treating cardiovascular disease in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0516] Also provided herein is a method for administering amino acid residues aa1-aa2 of SEQ. ID. NO. 632 to an individual in need thereof. 20 A method for treating cardiovascular disease in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0517] In some cases described in the above embodiments, the glucagon analog is administered when the cardiovascular disease is related to an ischemic event.

[0518] Also provided herein is a method for administering amino acid residues aa1-aa2 of SEQ. ID. NO. 303 to an individual in need thereof. 17 A method for treating cardiovascular disease in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0519] Also provided herein is a method for administering amino acid residues aa1-aa2 of SEQ. ID. NO. 303 to an individual in need thereof. 18 A method for treating cardiovascular disease in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0520] Also provided herein is a method for administering amino acid residues aa1-aa2 of SEQ. ID. NO. 303 to an individual in need thereof. 19 A method for treating cardiovascular disease in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0521] Also provided herein is a method for administering amino acid residues aa1-aa2 of SEQ. ID. NO. 303 to an individual in need thereof. 20 A method for treating cardiovascular disease in an individual in need thereof comprising administering a therapeutically effective amount of a glucagon analogue comprising administering to said individual ...

[0522] In some cases for the above-described embodiments, the glucagon analog is administered when the cardiovascular disease is related to an ischemic event.

[0523] In any of the above-described embodiments, the glucagon analog is modified with a surfactant X of formula 3-I:

[0524] [ka]

[0525] where: R 1a is, at each occurrence, a single bond, H, substituted or unsubstituted C1-C 30 present independently in the alkyl group, substituted or unsubstituted alkoxyalkyl group, substituted or unsubstituted aralkyl group, or steroid nucleus-containing moiety; R 1b , R 1c and R 1d is, at each occurrence, a single bond, H, substituted or unsubstituted C1-C 30 each independently present in an alkyl group, a substituted or unsubstituted alkoxyalkyl group, or a substituted or unsubstituted aralkyl group; W 1 exists independently at each occurrence in —CH2—, —CHO—, —(C═O), —(C═O)—O—, —(C═O)—NH—, —(C═S)—, —(C═S)—NH—, or —CHS—; W 2 is —O—, —CH2— or —S—; R 2 Each occurrence of represents a single bond to U, H, substituted or unsubstituted C1-C 30 Alkyl groups, substituted or unsubstituted alkoxyalkyl groups, or substituted or unsubstituted aralkyl groups, —NH2, —SH, C2-C4 alkenes, C2-C4 alkynes, —NH(C=O)-CH2-Br, —(CH2) m -maleimide, or -N3; n is 1, 2 or 3; m is 1 to 10.

[0526] In certain embodiments, the glucagon analog is modified with a surfactant X having the following structure:

[0527] [ka]

[0528] where: R 1a is a substituted or unsubstituted C1-C6 alkyl; and R 1b , R 1c and R 1d is H; W 1 is -(C=O)-NH-; W 2 is—O—; and R 2 is a single bond.

[0529] In some of the above-described embodiments, R 1a is C1-C 20 Alkyl groups, C8-C 20 Alkyl group, C 12 -C 18 Alkyl group or C 14 -C 18 It is an alkyl group.

[0530] 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 truncated or acylated at the N-terminus to produce analogs that exhibit reduced potency, partial agonist, and antagonist activity, as has been observed for 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 contents of which are incorporated herein by reference. For example, deletion of the first six residues of bPTH results in an agonistic 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 yield potent agonistic analogs. Other modifications to the N-terminus of the peptide, such as deletion or attachment of a D-amino acid such as D-Phe, can provide potent, 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 are within the scope of the contemplated embodiments described herein.

[0531] Also contemplated within the scope of the embodiments described herein are surfactants covalently attached to peptide analogs, where the native peptide has undergone any of the following modifications: acetylation, acylation, PEGylation, ADP-ribosylation, amidation, covalent attachment of lipids or lipid derivatives, covalent addition of phosphatidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cysteine ​​cross-links, formation of pyroglutamate, formylation, gamma-carboxylation, gamma-carboxylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, glycosylation, lipid addition, sulfation, glycosylation of glutamic acid residues, hydroxylation and ADP-ribosylation, selenoylation, sulfation, transfer RNA-mediated addition of amino acids to proteins such as arginylation and ubiquitination. See, e.g., Nestor, JJ Jr. (2007) Comprehensive Medicinal Chemistry II2:573-601; 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 AcadSci 663:48-62). Also contemplated within the scope of the embodiments described herein are branched or cyclic peptides that are branched or unbranched. Cyclic, branched polypeptides and branched cyclic peptides may result from post-translational natural processes or may be made by synthetic methods. In some embodiments, any of the peptide products described herein includes the peptide analogs described above, which are then covalently attached to an alkyl glycoside surfactant moiety.

[0532] Also contemplated within the scope of the embodiments presented herein are peptide chains substituted at appropriate positions by substitution of the analogs claimed herein, for example, by acylation on the linker amino acid at the ε-position of Lys with fatty acids such as octane, decane, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, 3-phenylpropanoic acid, etc., having saturated or unsaturated alkyl chains (Zhang, L. and Bulaj, G. (2012) Curr Med Chem 19:1602-1618). Non-limiting illustrative examples of such analogs are as follows:

[0533] [ka]

[0534] In further embodiments, the peptide chain is optionally substituted at appropriate positions with a spacer and hydrophobic moiety, such as a steroid nucleus, for example, a cholesterol moiety, by reaction on a linker amino acid, such as the sulfhydryl group of Cys. In some such embodiments, the modified peptide further comprises one or more PEG chains. Non-limiting examples of nanoencapsulation are as follows:

[0535] [ka]

[0536] In addition to the 20 standard amino acids, a vast number of "non-standard amino acids," or artificial amino acids, exist and are known in the art and have been incorporated into the compounds described herein as described above. Other non-standard amino acids have been modified with reactive side chains for 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 approach, a transfer ribonucleic acid / tRNA synthetase pair was engineered and encoded in an expression plasmid with 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 then reacted with a functionalized surfactant or a PEG polymer bearing an acetylene moiety in the presence of a reducing agent and copper ions to promote an organic reaction known as the "Huisgen [3+2] cycloaddition." Similar reaction sequences using the reagents described herein containing acetylene-modified alkyl glycosides or PEG-modified glycosides yield PEGylated or alkyl glycoside-modified peptides. For peptides of less than about 50 residues, standard solid-phase synthesis is used to attach the reactive amino acid residues to the desired positions in the chain. Surfactant-modified peptides and / or proteins exhibit a different spectrum of pharmacological and medicinal properties than peptides modified solely by PEG attachment.

[0537] Those skilled in the art will recognize that numerous permutations of peptide analogs can be made that, if they have surfactant moieties incorporated into the amino acid sequence, will possess the desired attributes of the surfactant-modified peptide products described herein.

[0538] <Specific definition> As used herein, "a" or "an" means one or more. As used in the claims, when used in conjunction with the word "comprising," the words "a" or "an" mean one or more. As used herein, "another" means at least two or more.

[0539] As used herein, the one- and three-letter abbreviations for the various common amino acids are as recommended in Pure Appl. Chem 31, 639-645 (1972) and 40, 277-290 (1974) and conform to the provisions of 37 CFR § 1.822 (55 FR 18245, May 1, 1990). Unless specified as D- or DL, the abbreviation refers to an L-amino acid. Certain amino acids, both natural and artificial, are achiral, e.g., glycine, α-amino-isobutyric acid (Aib). All peptide sequences are presented with the N-terminal amino acid on the left and the C-terminal amino acid on the right.

[0540] An "alkyl" group refers to an aliphatic hydrocarbon group. Reference to an alkyl group includes "saturated alkyl" and / or "unsaturated alkyl," i.e., alkene or alkyne. Alkyl groups, whether saturated or unsaturated, include branched, straight-chain, or cyclic groups. Alkyl groups are optionally substituted by including substituents, including, but not limited to, oxo, halogen, aryl, cycloalkyl, hydrophobic natural groups such as steroids, aralkyl chains (including alkoxyaryl), and alkyl chains containing acyl moieties. In some embodiments, an alkyl group is linked to the Nα-position of a residue in a peptide (e.g., Tyr or Dmt). This class is referred to as N-alkyl and includes C1-C10 straight-chain or branched alkyl groups, or aryl-substituted alkyl groups such as benzyl and phenylethyl. In some embodiments, the alkyl moiety is a 1-alkyl group present at the glycosidic linkage to the saccharide moiety (typically at the 1-position of, for example, glucose). One such alkyl group is a C1-C30 alkyl group.

[0541] An "aryl" group refers to an aromatic ring, in which each of the atoms forming the ring is a carbon atom. Aryl rings described herein include rings having 5, 6, 7, 8, 9, or 10 or more carbon atoms. Aryl groups are optionally substituted with substituents selected from halogen, alkyl, acyl, alkoxy, alkylthio, phenyl, dialkylamino, and carboxyl ester, such as cyano. Examples of aryl groups include, but are not limited to, phenyl and naphthalenyl.

[0542] The term "acyl" refers to a C1-C20 acyl chain, which may include a straight aliphatic chain, a branched aliphatic chain, a chain containing a cyclic alkyl moiety, a hydrophobic natural product such as a steroid, an aralkyl chain, or an alkyl chain containing an acyl moiety.

[0543] The term "steroid nucleus" refers to the core of a steroid containing four fused rings designated A, B, C, and D as shown below.

[0544] [ka]

[0545] Examples of steroid containing moieties include, but are not limited to, cholesterol, and the like.

[0546] As used herein, a "therapeutic composition" can include a mixture with an aqueous or organic carrier or excipient, and can be formulated with conventional non-toxic, pharmaceutically acceptable carriers for tablets, pellets, capsules, lyophilisates, suppositories, solutions, emulsions, suspensions, or other forms suitable for use. In addition to those disclosed above, carriers can include alginate, collagen, glucose, lactose, mannose, gum arabic, 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 manufacture of formulations in solid, semi-solid, or liquid form. Additionally, auxiliary stabilizing, thickening, or coloring agents can be used, such as dihydroxyacetone (triulose), to stabilize dried drugs.

[0547] For example, as used herein, a "pharmaceutically acceptable carrier" or "therapeutically effective carrier" may be aqueous or non-aqueous (solid), e.g., alcoholic or oily, or a mixture thereof, and may include surfactants, emollients, lubricants, stabilizers, dyes, fragrances, preservatives, acids or bases for adjusting pH, solvents, dispersants, gelling agents, moisturizers, stabilizers, humectants, time-release agents, humectants, or other components commonly included in a particular form of pharmaceutical composition. Pharmaceutically acceptable carriers are well known to those skilled in the art and include, for example, aqueous solutions such as water or physiologically buffered saline, or other solvents or vehicles such as glycols, glycerol, and oils such as olive oil or injectable organic acid esters. Pharmaceutically acceptable carriers can include, for example, specific inhibitors, carbohydrates such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, other stabilizers, physiologically acceptable compounds that act to stabilize or increase absorption of the excipient.

[0548] Pharmaceutical compositions can also include other pharmaceutically acceptable auxiliary substances as desired to approximate physiological conditions; such "substances" include, but are not limited to, pH adjusting and buffering agents, tonicity adjusting agents, and the like, e.g., sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, etc. Additionally, peptide, or variant thereof, suspensions can contain lipid-protecting agents that protect lipids from free radical and lipid peroxidation damage during storage. Lipophilic free-radical quenchers, such as alpha-tocopherol and water-soluble iron-specific chelators, such as ferrioxamine, are suitable.

[0549] As used herein, a "surfactant" is a surface active agent that modifies the interfacial tension of water. Typically, surfactants have one lipophilic and one hydrophilic group or region in the molecule. Broadly, the groups include soaps, detergents, dispersants, dispersing and wetting agents, and preservatives. More specifically, surfactants include stearyl triethanolamine, sodium lauryl sulfate, sodium taurocholate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glycerol monostearate; and hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, polyethylene glycol (PEG), carboxymethylcellulose, sodium, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose, or alkylglycosides. In some embodiments, the surfactant is a nonionic surfactant (e.g., an alkylglycoside surfactant). In some embodiments, the surfactant is an ionic surfactant.

[0550] As used herein, "alkyl glycoside" refers to any sugar linked by a bond to any hydrophobic alkyl, as known in the art. The hydrophobic alkyl can be selected from any desired size depending on the desired hydrophobicity and hydrophilicity of the saccharide moiety. In one embodiment, the alkyl chain ranges from 1 to 30 carbon atoms; alternatively, it ranges from 6 to 16 carbon atoms.

[0551] As used herein, "saccharide" includes monosaccharides, oligosaccharides, or polysaccharides in linear or cyclic configurations. Oligosaccharides are saccharides with two or more monosaccharide residues. Some examples of the many possible saccharides suitable for use in functionalized form include glucose, galactose, maltose, maltotriose, maltotetraose, sucrose, trehalose, etc.

[0552] As used herein, "sucrose esters" are sucrose esters of fatty acids. Sucrose esters can take many forms due to the eight hydroxyl groups in sucrose and the many fatty acid groups available for reaction, from acetates up to the larger, more bulky fats that can be reacted with sucrose. This flexibility means that many products and functionalities can be tailored based on the fatty acid moiety used. Sucrose esters have food and non-food uses, particularly as surfactants and emulsifiers, with growing applications in pharmaceuticals, cosmetics, detergents, and feed additives. They are biodegradable, non-toxic and non-irritating to the skin.

[0553] As used herein, a "suitable" alkyl glycoside means one that is non-toxic and non-ionic. In some instances, a suitable alkyl glycoside reduces immunogenicity or aggregation and increases the bioavailability of a compound when administered with the compound via ophthalmic, nasal, nasolacrimal, sublingual, buccal, or inhalation routes, or by injection routes such as subcutaneous, intramuscular, or intravenous routes. Suitable compounds can be determined using methods known in the art and described in the Examples.

[0554] A "linker amino acid" is any natural or artificial amino acid containing a reactive functional group (de Graaf, AJ et al. (2009) Bioconjug Chem 20:1281-1295) that can be used to form a covalent bond with a functionalized surfactant. By way of example, in some embodiments, the linker amino acid is Lys or Cys, which has the reactive functional group -NH; or cysteine, which has the reactive functional group -SH; or Asp or Glu, which has the reactive functional group -C(=O)-OH. By way of example, in other particular embodiments, the linker amino acid is any amino acid containing a reactive functional group such as -OH, -N, haloacetyl, or an acetylene group that can be used to form a covalent bond with an appropriately functionalized surfactant.

[0555] As used herein, a "functionalized surfactant" is a surfactant that includes a reactive group suitable for covalent bonding with a linker amino acid. By way of example, in some embodiments, the functionalized surfactant includes a carboxylic acid group (e.g., at the 6-position of a monosaccharide) as a reactive group suitable for covalent bonding with a linker amino acid. By way of example, in some embodiments, the functionalized surfactant includes an -NH group, an -N group, an acetylene group, a haloacetyl group, an -O-NH group, or a -(CH-) group. m -maleimide group, for example, at the 6-position of a monosaccharide (as shown in Scheme 6), which allows for covalent attachment with a suitable linker amino acid. In some embodiments, as described herein, the functionalized surfactant is a compound of Formula IV:

[0556] As used herein, the term "peptide" refers to any peptide containing two or more amino acids. The term "peptide" includes short peptides (e.g., peptides containing between 2 and 14 amino acids), medium-length peptides (15-50), or long-length peptides (e.g., proteins). The terms peptide, medium-length peptide, and protein may be used interchangeably herein. As used herein, the term "peptide" is intended to mean a polymer composed of amino acid residues associated with naturally occurring structural variants and synthetic, non-naturally occurring analogs linked via peptide bonds. Synthetic peptides can be synthesized, for example, using automated peptide synthesizers. Peptides may contain amino acids other than the 20 gene-encoded amino acids. "Peptides" include those modified by either natural processes, such as processing and other post-translational modifications, as well as by chemical modification techniques. Such modifications are described in basic texts and more detailed monographs and are well known to those of skill in the art. It is recognized that the same type of modification may be present to the same or varying degrees at several sites in a given peptide. Also, a given peptide may, in some embodiments, contain more than one type of modification. Modifications can occur anywhere in a peptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini.

[0557] Accordingly, and contemplated within the scope of the embodiments described herein are surfactants covalently attached to peptides, wherein the peptides have been modified by modifications including, for example, acetylation, acylation, pegylation, ADP-ribosylation of lipids or lipid derivatives, amidation, covalent addition of phosphatidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cysteine ​​cross-links, pyroglutamate formation, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, glycosylation, lipid attachment (sulfation) of glutamic acid residues, hydroxylation and transfer RNA-mediated addition of amino acids to proteins such as ADP-ribosylation, selenoylation, sulfation, arginylation, and 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 663:48-62). Also contemplated within the embodiments described herein are branched or unbranched branched or cyclic peptides. Cyclic, branched polypeptides and branched cyclic peptides may result from post-translation natural processes or may be made by synthetic methods.

[0558] The term "peptide" includes peptides or proteins containing natural and artificial amino acids or analogs of natural amino acids. As used herein, peptide and / or protein "analogs" include artificial amino acids based on natural amino acids, such as tyrosine analogs, including p-substituted tyrosines, o-substituted tyrosines, and m-substituted tyrosines, where the substituents on tyrosine are acetyl, benzoyl, amino, hydrazine, hydroxylamine, thiol, carboxy, methyl, isopropyl, C2-C 20 Examples of analogs include straight-chain or branched hydrocarbons, mono-, 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-aryl-tyrosine, 4-propyl-tyrosine, Cα-methyl-tyrosine, etc. Examples of lysine analogs include ornithine (Orn), homo-lysine, Cα-methyl-lysine (CMeLys), etc. Examples of phenylalanine analogs include, but are not limited to, meta-substituted phenylalanine, where the substituents include methoxy groups, C1-C20 alkyl groups, e.g., methyl groups, aryl groups, acetyl groups, etc. Specific examples include, but are not limited to, 2,4,6-trimethyl-L-phenylalanine (Tmp) and 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), fluorinated phenylalanine, isopropyl-phenylalanine, p-azido-phenylalanine, p-acyl-phenylalanine, p-benzoyl-phenylalanine, p-iodo-phenylalanine, p-bromophenylalanine, p-amino-phenylalanine, and isopropyl-phenylalanine. Among the vast array of non-standard or artificial amino acids known in the art and used in the design of peptide analogs are C-alpha-disubstituted amino acids such as Aib, Cα-diethylglycine (Deg), aminocyclopentane-1-carboxylic acid (Ac4c), and aminocyclopentane-1-carboxylic acid (Ac5c). Such amino acids often lead to constrained conformations that bias toward alpha helical structures (Kaul, R. and Balaram, P. (1999) BioMed Chem 7:105-117). Additional examples of such artificial amino acids useful in analog design include homo-arginine (Har). In certain instances, reduced amide bond substitution leads to improved protection from enzymatic destruction or modifies receptor binding.As an example, a Tic-Phe dipeptide-based bond with a reduced amide bond between the residues (Tic-Ψ[CH2-NH]-Ψ-Phe) reduces enzymatic degradation. Thus, contemplated within the scope of the embodiments described herein are surfactants covalently attached to peptides containing the modified amino acids and / or peptide analogs described above. Specific artificial amino acids are shown below.

[0559] [ka]

[0560] As used herein, an "opioid peptide" is a short sequence of amino acids that binds to opioid receptors in the body. In some embodiments, the opioid peptide is an endogenous peptide, such as an endorphin, enkephalin, endomorphin, dermorphin, or the like. In some embodiments, the opioid peptide is derived from an endogenous morphine-like peptide (e.g., a pseudopeptide, a suppressed peptide, an alpha-methyl analog, etc.). In some embodiments, the opioid peptide is exogenous and / or synthetic and includes modified and / or artificial amino acids that mimic the effects of the opioid peptide.

[0561] As used herein, the term "variant" is understood to mean a peptide that differs from a reference peptide but retains essential properties. A typical variant of a peptide differs from another reference peptide in its amino acid sequence. Generally, the differences are limited, so that the sequences of the reference peptide and the variant are similar overall and, in many regions, identical. A variant and a reference peptide may differ in amino acid sequence by one or more substitutions, additions, or deletions, in any combination. The substituted or inserted amino acid residues may or may not be those encoded by the genetic code. Artificially generated variants of peptides can be made by mutagenesis techniques, direct synthesis, and other suitable recombinant methods.

[0562] Reference will now be made in detail to the various embodiments and specific applications of the covalently modified peptides and / or proteins described herein. While the covalently modified peptides and / or proteins are described in conjunction with various embodiments and applications, it will be understood that such embodiments and applications are illustrative and are not intended to limit the scope of the embodiments described herein. Additionally, throughout this disclosure, various patents, patent applications, websites, and publications are referenced, and unless otherwise indicated, each is incorporated herein by reference for its relevant disclosure.

[0563] <Peptide> There are many important roles played by peptides in the body, and several commercial opportunities have been exploited (Nestor, JJ Jr. (2009) Current Medicinal Chemistry 16:4399-4418; Stevenson, CL (2009) Curr Pharm Biotechnol 10:122-137). However, even these recognized targets and products continue to suffer from deficiencies in duration of action and bioavailability (Tyndall, JD et al. (2005) Chem Rev 105:793-826). In some embodiments, the improved peptides described herein provide longer duration of action and / or bioavailability and / or therapeutic efficacy compared to currently available commercial products. Some illustrative examples of peptides that represent attractive commercial targets for analog design (agonists and antagonists) for clinical development include, for example, class B members, G protein-coupled receptor (GPCR) ligands and related peptides ("secretin family"): secretin, parathyroid hormone (PTH), parathyroid hormone-related protein (PTHrP), glucagon, glucagons such as proteins-1 and -2 (GLP-1, GLP-2), glucose-dependent insulin-like polypeptide (GIP), oxyntomodulin, pituitary adenylate cyclase-activating peptide (PACAP), vasoactive intestinal peptide (VIP), amylin (and analogs such as pramlintide, devalintide, etc.), calcitonin (and analogs such as salmon calcitonin, elcatonin, etc.), calcitonin gene-related peptide (CGRP), adrenomedullin, corticotropin-releasing factor family members (CRF, Xerecept, etc.), These include synthetic analogs of peptides (e.g., urocortin) that may be improved as clinical products by further modification using the methods described herein. Also contemplated within the scope of the embodiments presented herein are the opioid peptide family; such peptides would benefit from the peptide modification methods described herein to confer increased duration of action and increased specificity.By way of example, peptide families such as endomorphins, dynorphins, enkephalins, dermorphins, and casomorphins offer attractive therapeutic approaches for the treatment of pain, addiction, etc. Additional attractive peptide targets for modification to yield the peptide products described herein are hypothalamic hormones, such as gonadotropin-releasing hormone and its analogs (e.g., nafarelin, goserelin, triptorelin, leuprorelin, fertirelin, histrelin, buserelin, ganirelix, cetrorelix, degarelix, deslorelin, etc.), adrenocorticotropic hormone, somatostatin (e.g., octreotide, lanreotide, vapreotide, etc.), thyrotropin-releasing hormone, growth hormone-releasing hormone, and neurotensin. Further examples of attractive commercial targets are pituitary hormones and analogs such as vasopressin (desmopressin, etc.), oxytocin and analogs, thyroid stimulating hormone, prolactin, growth hormone, luteinizing hormone, vesicle stimulating hormone, alpha-melanocyte stimulating hormone analogs (melanotan analogs), and analogs thereof. Growth factors are an important class of molecules that can be advantageously modified using the methods described herein to improve pharmaceutical candidates, such as insulin (and analogs such as lispro, levemir, and glargine), insulin-like growth factor I (IGF-I or somatomedin C), nerve growth factor (NGF), fibroblast growth factors (FGFs; FGF-18, FGF-20, FGF-21, etc.), keratinocyte growth factor (KGF), and vascular endothelial growth factor (VEGF). Particularly attractive targets are peptides that regulate gut function and appetite but have a short duration of action (some of which are mentioned above), including, but not limited to, ghrelin, pancreatic peptide, peptide YY, neuropeptide Y, cholecystokinin (such as sincalide), melanocortin, and the like.Additional targets that would benefit from the methods described herein are pro-inflammatory adipose tissue products associated with obesity, such as leptin (and related analogs, such as OB-3 peptide), adipokines, adiponectin, chemerin, visfatin, nesfatin, resistin, tumor necrosis factor alpha, chemokines, monocyte chemoattractant protein-1 (MCP-1), omentin, interleukins, etc. Important targets that would benefit from improved pharmaceutical and pharmacological behavior are proteins that regulate immune function, including, but not limited to, members of the interferon family (interferons alpha, beta, gamma, kappa, omega, including IL-10, IL-19, IL-20, IL-22, IL-24, IL-26, etc.), thymopentin, thymosin alpha 1, etc. Important peptide products that circulate or regulate blood clotting may be improved by the methods described herein, thereby achieving increased duration of action, such as bivalirudin (Angiomax), eptifibatide (Integrelin), atrial natriuretic peptide (ANP, ularitide), brain natriuretic peptide, C-type natriuretic peptide, b-type natriuretic peptide (nesiritide), angiotensin, hematopoietic inhibitors, rotigaptide, thrombospondin, and the like. Peptides that stimulate stem cell proliferation and differentiation are important agents that may be improved by the modifications described herein. For example, erythropoietin, hematide, thrombopoietin, macrophage colony-stimulating factor (M-colony-stimulating factor), leukemia inhibitory factor (LIF), interleukin 6 (IL-6), granulocyte colony-stimulating factor (G-colony-stimulating factor), granulocyte-macrophage colony-stimulating factor (GM-colony-stimulating factor), and the like are contemplated as peptides suitable for covalent attachment to surfactants (e.g., alkyl glycoside surfactants). Neurotrophic factors are another small class of proteins that would greatly benefit from the modifications described herein to increase their duration of action and potency.For example, glial cell-derived neurotrophin (GDNF) and its family (neurturin, artemin, parsfin) (neurotrophins such as nerve growth factor (NGF), BDNF, and others). Proinflammatory and pain-inducing peptides are important peptide targets that can be improved by the modifications described herein. For example, inhibitors, particularly antagonists, of bradykinin (or its release (e.g., ecallantide)) and substance P provide important therapeutic targets (e.g., icatibant) and are amenable to covalent attachment of surfactants (e.g., alkylglycoside surfactants). Inhibitors of viral fusion (fuzeon), protein maturation (protease inhibitors), or integration suffer from a short duration of action. Modification of such inhibitors by covalent attachment of surfactants (e.g., alkylglycoside surfactants) allows for a longer duration of action. Many proteases have been found to be involved in diseases and could be further improved by the modifications and targeting described herein. Moreover, inhibitors of their action have reached the clinic (Abbenante, G. and Fairlie, DP (2005) Med Chem 1:71-104), and such targets are contemplated within the scope of the embodiments described herein. Additional natural peptide products and their analogs, such as conotoxin peptides for pain relief, antibacterial defensins, and the like, also suffer from lack of bioavailability and short duration of action in physiological fluids and therefore would benefit from the peptide modifications described herein. Those skilled in the art will recognize many additional commercially important peptides that are amenable to the modifications described herein to provide increased duration of action and bioavailability. Such peptides are also contemplated within the scope of the present disclosure.

[0564] Amino- or carboxyl-terminal modifications may be optionally introduced into the peptides (Nestor, JJ Jr. (2009) Current Medicinal Chemistry 16:4399-4418). For example, the peptides may be truncated or acylated at the N-terminus to result in peptides exhibiting reduced potency, partial agonist, and antagonist activity, as seen for 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 contents of which are incorporated herein by reference. Other modifications to the N-terminus of the peptide, such as the deletion or attachment of D-amino acids such as D-phenylalanine, can also provide potent, 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 are within the scope of the contemplated embodiments described herein.

[0565] In addition to the 20 standard amino acids, there are a vast number of "non-standard amino acids," or amino acids known in the art that can be incorporated into the compounds described herein, as described above. Other non-standard amino acids have been modified with reactive side chains for conjugation (Gauthier, MA and Klok, HA (2008) Chem Commun (Camb) 2591-2611; de Graaf, AJ et al. (2009) Bioconjug Chem 20:1281-1295). One approach has been to develop transfer ribonucleic acid / tRNA synthetase pairs codon-encoded by amber suppressors in expression plasmids (Deiters, A et al. (2004) Bio-org. Med. Chem. Lett. 14, 5743-5). For example, p-azidophenylalanine was incorporated into a peptide and then reacted with a functionalized surfactant or a PEG polymer bearing an acetylene moiety in the presence of a reducing agent and copper ions to promote an organic reaction known as the Huisgen [3+2] cycloaddition. Similar reaction sequences using the reagents described herein, including acetylene-modified alkyl glycosides or PEG-modified glycosides, result in PEGylated or alkyl glycoside-modified peptides. For peptides of less than about 50 residues, standard solid-phase synthesis is used to attach the reactive amino acid residues to the desired positions in the chain. Surfactant-modified peptides and / or proteins exhibit a different spectrum of pharmacological and medicinal properties than peptides modified solely by PEG attachment.

[0566] <Intermediate> In one embodiment, provided herein are intermediates and / or reagents containing a surfactant moiety and a reactive functional group capable of forming a single bond with a reactive functional group on a natural or artificial amino acid. These intermediates and / or reagents can be used to improve the bioavailability of peptides and / or proteins used in human and veterinary diseases, as well as the pharmacokinetic and / or pharmacological behavior of the drug. Covalent attachment of such intermediates and / or reagents to functional groups on the side chains of amino acids, for example, to the epsilon-amino functional group of Lys, the sulfhydryl group of Cys, or a sulfhydryl group at the amino or carboxy terminus of a peptide and / or protein target, enables the synthesis of the peptide products described herein. In a specific embodiment, the nonionic surfactant moiety is a monosaccharide or disaccharide having an O-alkyl glycosidic substitution, where the glycosidic bond has an alpha or beta structure. In a specific embodiment, the O-alkyl chain is C1-C 20 Alkyl chain, or C6-C 16 From alkyl chains.

[0567] In another embodiment, provided herein are intermediates and / or reagents comprising nonionic surfactant moieties with specific alkylglycosidic linkages that mimic O-alkylglycosidic linkages and reactive functional groups capable of forming single bonds with reactive functional groups on natural or artificial amino acids. Such intermediates and / or reagents include S-linked or N-linked alkyl chains and exhibit modified chemical and / or enzymatic stability compared to O-linked alkylglycosidic linked products.

[0568] In some embodiments, the intermediates and / or reagents provided herein are compounds wherein the hydrophilic group is modified glucose, galactose, maltose, glucuronic acid, diglucuronic acid, maltouronic acid, etc. In some embodiments, the hydrophilic group is glucose, maltose, glucuronic acid, diglucuronic acid, or maltauronic acid, and the hydrophobic group is C1-C 20 It is an alkyl or aralkyl chain. In some embodiments, the glycosidic bond to the hydrophobic group is in the alpha configuration, and in some, the bond is in the beta configuration at an anomeric center on the saccharide.

[0569] In some embodiments, the hydrophilic group is glucose, maltose, glucuronic acid, diglucuronic acid, or maltauronic acid, and the hydrophobic group is C1-C 20 It is an alkyl chain or an aralkyl chain.

[0570] In some embodiments, the intermediates and / or reagents provided herein include surfactants that include reactive functional groups that are carboxylic acid groups, amino groups, azides, aldehydes, maleimides, mercapto groups, hydroxylamino groups, alkynes, or the like.

[0571] In some embodiments, the intermediates and / or reagents are O-linked alkyl glycosides with one of the hydroxyl groups modified to become a carboxylic acid or amino functionality. In some embodiments, the reagent is a 1-O-alkylglucuronic acid in the alpha or beta configuration, where the alkyl chain is C1 to C20 in length. In some such embodiments, the alkyl group is C6 to C18 in length. In some such embodiments, the alkyl group is C6 to C16 in length.

[0572] In some embodiments, the reagent comprises a 1-O-alkyldiglucuronic acid in the alpha or beta configuration, where the alkyl chain is C1 to C20 in length. In some such embodiments, the alkyl group is C6 to C16 in length.

[0573] In some embodiments, the reagent is an S-linked alkyl glycoside in the alpha or beta configuration, with one of the hydroxyl groups modified to be a carboxylic acid or amino functionality.

[0574] In some embodiments, the reagent is an N-linked alkyl glycoside in the alpha or beta configuration, with one of the hydroxyl groups modified to be a carboxylic acid or amino functionality.

[0575] In yet another embodiment, there are provided herein peptide and / or protein products comprising covalently linked alkyl glycosides that have acceptable properties for use in human and veterinary disease. Scheme 1 lists exemplary nonionic surfactants that can be modified to provide reagents and / or intermediates useful in the synthesis of surfactant-modified peptide products described herein.

[0576] [ka]

[0577] In some embodiments, the covalently modified peptides and / or proteins described herein incorporate surfactant moieties into the peptide structure. In certain embodiments, the covalently modified peptides and / or proteins described herein incorporate nonionic surfactants of the alkyl, alkoxyalkyl, or aralkyl glycoside class. Alkyl glycosides are important commodities, widely used in the food, hospitality, 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 their production at very low cost (Park, DW et al. (2000) Biotechnology Letters 22:951-956). These alkyl glycosides can be further modified to yield intermediates for the synthesis of the covalently modified peptides and / or proteins described herein. Thus, it is known that 1-dodecyl beta-D-glucoside is preferentially oxidized at the 6-position to give the corresponding glucuronic acid analog in high yield when using unprotected material and platinum black catalyst in the presence of oxygen (van Bekkum, H. (1990) Carbohydrates as Organic Raw Materials 289-310). Additional chemoselective methods for the oxidation of primary alcohols at the 6-position of alkyl glucosides are available. For example, the use of catalytic amounts of 2,2,6,6-tetramethyl-1-piperidyloxyl (TEMPO) with the stoichiometry of the organic oxidant [bia(acetoxy)iodo]benzene (BAIB) (De Mico, A. et al. (1997) J Org Chem 1997:6974-6977) afforded remarkable yields of nucleoside-5'-carboxylic acids by oxidation of the primary hydroxyl (Epp, JB and Widlanski, TS (1999) J Org Chem 64:293-295). This oxidation is chemoselective toward the primary hydroxyl, even when the secondary hydroxyl is unprotected (Codee, JD et al. (2005) J Am Chem Soc 127:3767-3773).Similarly, 1-dodecyl β-D-glucopyranoside and 1-tetradecyl β-D-glucopyranoside were oxidized to the corresponding uronic acids (1-dodecyl β-D-glucuronic acid and 1-tetradecyl β-D-glucuronic acid) by oxidation with TEMPO using KBr and sodium hypochlorite as stoichiometric oxidants in water (Milkereit, G. et al. (2004) Chem Phys Lipids 127:47-63). A mild oxidation using (diacetoxyiodo)benzene (DAIB·alk·BAIB) is given in the Examples. Certain of these glucuronic acid intermediates are commercially available (e.g., octyl bD-glucuronic acid; Carbosynth, MO 07928) and, as shown, are widely prepared by routine methods or by invention (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). Scheme 2 illustrates certain functionalized surfactant intermediates containing a —COOH group as a reactive functional group used to prepare the intermediates and / or reagents described herein.

[0578] [ka]

[0579] Similarly, aralkyl glycosides (including alkoxyalkyl) can form the basis of closely related nonionic surfactant reagents. For example, 4-alkoxyphenyl β-D-glucopyranosides are readily synthesized by the reaction of penta-O-acetyl β-D-glucose with 4-alkoxyloxyphenols in the presence of boron trifluoride etherate. As described above and in the Examples, deacetylation using trimethylamine in methanol / water affords alkoxyaryl glucuronic acid reagents suitable for forming reagents and peptides after selective oxidation (Smits, E. et al. (1996) J Chem Soc, Parkin Trans I 2873-2877; Smits, E. et al. (1997) Liquid Crystal 23:481-488).

[0580] [ka]

[0581] Intermediates of the glucuronic acid class are easily activated by standard coupling agents for attachment to amino acid side chains (e.g., the side chain of Lys). Thus, Fmoc-Lys-O-Tm (trimethylsilyl = TMS) can be reacted with octyl beta-D-glucuronic acid in the presence of a coupling agent and subsequently hydrolyzed in an aqueous workup to yield Fmoc-Lys (1-octyl β-D-glucuronic acid amide), as shown in Scheme 4. This reagent can be used for coupled blast-phase synthesis of peptides using standard coupling protocols when it is desired to incorporate a surfactant moiety near the N-terminal region of the molecule. The second hydroxyl group can be left unprotected due to the extremely high reactivity of the Lys amino function, or it can be protected by perchlorosilation. 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. Scheme 4 illustrates the formulation of the reagents described herein.

[0582] [ka]

[0583] In some embodiments, reagents and / or intermediates for the formulation of active peptide products described herein include a family of linker amino acids that are modified into surfactants for attachment to synthetic peptide products. Thus, in one embodiment, the peptide products described herein are synthesized in a linear fashion, in which a functionalized surfactant is added to a linker amino acid that is reversibly protected by a functional group on the side chain of the linker amino acid (e.g., the amino group of a lysine residue) to yield a unique reagent (as shown in Scheme 4), which is incorporated into a growing peptide chain, and the remainder of the peptide is then synthesized by adding amino acids to additional cysteine ​​residues. Suitable protecting groups for the synthesis of modified peptides and / or proteins described herein are described, for example, in T.W. Green, P.G. M. Buts, Protective Groups in Organic Synthesis, Wiley-Interscience, New York, 1999, pp. 503-507, 736-739, the disclosure of which is incorporated herein by reference.

[0584] In another embodiment, the peptide products described herein are synthesized by the covalent addition of a functionalized surfactant to an intact peptide via an appropriate functional group on a linker amino acid in the peptide chain.

[0585] Alternatively, the functionalized surfactant can be added to a deprotected linker amino acid side chain during solid-phase peptide synthesis. As an example, an alkylglucuronyl group can be added directly to a linker amino acid side chain (e.g., a deprotected Lys side chain) during solid-phase peptide synthesis. For example, the use of Fmoc-Lys(Alloc)-OH as a subunit provides vertical protection that can be removed while the peptide is still on the resin. Therefore, deprotection of the leasing side chain using Pd / thiobarbital or other allocative deprotection recipes allows exposure of the amino group for attachment to a protected acyl or unprotected 1-octyl β-D-glucuronic acid unit. Final deprotection with a low percentage of CF3CO2H (TFA) cleavage cocktail then delivers the desired product. While the glycosidic bond is labile to strong acids, experience here and by others indicates that it is relatively stable to low percentages of TFA cleavage. Alternatively, acyl protection (e.g., acetyl, Ac; benzoyl, Bz) or trialkylsilyl protection on the sugar OH function may be used to provide increased protection to the glycosidic bond. Subsequent deprotection with base (NH2NH2 / MeOH; NH3 / MeOH, NaOMe / MeOH) yields the desired deprotected product. Scheme 4 illustrates the reagents described herein. Scheme 5 illustrates the peptide intermediates described herein.

[0586] [ka]

[0587] As shown below in Scheme 6, additional reagents can be generated by modification of the 6-position functionality to provide various means of attachment to amino acid side chain functionalities. Thus, an amino substituent can be used for attachment to Asp or Glu side chains. Azido or alkyne substituents can be used for attachment to artificial amino acids containing complementary acceptors for Huisgen 3+2 cycloaddition (Gauthier, MA and Klok, HA (2008) Chem Commun (Cam) 2591-2611). Aminoxy or aldehyde functionalities can be used to link aldehydes (i.e., oxime linkages) or amino functionalities (i.e., contracting alkylations), respectively. Maleimide or —NH(C═O)—CH—Br functionalities can be chemoselectively attached via Cys or other SH functionalities. These types of attachment strategies are advantageous when used in conjunction with the reagents described herein. Functional group interconversions are widely practiced in organic synthesis, and comprehensive lists of multiple routes to each of the functional group modifications listed herein are available (Larock, R.C. (1999) "Comprehensive Organic Transformations", VCH Publishers, New York).

[0588] Thus, the primary hydroxyl on position 6 of octyl 1-β-D-glucoside is converted to an azide by activation and substitution with azide anion, a reaction similar to that used in carbohydrate chemistry (e.g., by tosylation followed by NaN). The corresponding azide is converted to an amino function by reduction with thioacetic acid in pyridine (Elofsson, M. et al. (1997) Tetrahedron 53:369-390) or by analogous methods of amino group generation (Stangier, P. et al. (1994) Liquid Crystal 17:589-595). Access to acetylene, aminoxy, and aldehyde moieties is best performed on the triacetoxy forms available from commercially available glucosides by treatment with AcO followed by mild hydrolysis of the primary amine. This 6-hydroxy form can be selectively oxidized to the aldehyde or activated as the tosylate or triflate and displaced with NHOH or sodium acetylide. The maleimide bond may again be present via a carbon bond as shown, preferably an O or amide bond, by displacement of the activated hydroxyl, or by coupling of an amino-linked glucuronic acid derivative with maleimide reagents well known in the art. Additional functional group transformations are well known to those skilled in the art of pharmaceutical chemistry and are within the scope of the embodiments described herein.

[0589] Also contemplated within the scope of the synthetic methods described herein are surfactants in which the sugar and hydrophobic chain are covalently attached via an alpha-glycosidic bond. Synthetic routes to primarily alpha-linked glycosides are well known in the art and typically start from peracetylated sugars and use an acid catalyst (e.g., SnCl, BF, or HCl) to produce alpha-glycosylation (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). Similar synthetic routes exist 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 then proceed as described above, leading to the 6-carboxylic acid, etc., for subsequent generation of the corresponding α-linked reagent.

[0590] Scheme 6 lists certain compounds and reagents useful in the synthesis of covalently modified peptides and / or proteins described herein. Single letter abbreviations using the full names for amino acids are used.

[0591] [ka]

[0592] Many alkyl glycosides can be synthesized by known procedures, e.g., 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. Pat. Nos. 3,219,656 and 3,839,318, or enzymatically, e.g., 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 coupling to non-natural amino acids such as Ser can be performed on Fmoc-Ser-OH using peracetylglucose to yield N-Fmoc-4-O-(2,3,4,6-tetra-O-acetyl-β-D-glucopyranodyl)-L-serine. This material is selectively deprotected at the primary carbon atom (position 6) and selectively oxidized using TEMPO / BAIB as described above to yield the corresponding 6-carboxyl function, which can be coupled to lipophilic amines to generate a new class of nonionic surfactants and reagents (Scheme 7).

[0593] [ka]

[0594] The linkage between the hydrophobic alkyl and the hydrophilic sugar can involve, among other possibilities, a glycoside, a thioglycoside, an amide (Carbohydrate as Organic Raw Materials, F.W. Lichtenthaler, ed., VCH Publishers, New York, 1991), a ureid (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 an ester linkage (Sugar Esters: Preparation and Application, J.C. Colbert, ed., Noyes Data Corp., New Jersey, (1974)).

[0595] Examples of alkyl glycosides useful for modification to the reagents described herein or formulation of products from which they may be selected include the following: alkyl glycosides, octyl-, nonyl-, decyl-, undecyl-, dodecyl-, tridecyl-, tetradecyl-, pentadecyl-, hexadecyl-, heptadecyl-, and octadecyl-D-maltocoside, -glucoside, or -sucrose ester (i.e., sucrose ester) (synthesized according to Koeltzow and Urfer; Anatrace, Maumee, Ohio; Calbiochem, San Diego, Calif.; Fluka Chemie, Switzerland); Heptyl, octyl, dodecyl, tridecyl, and tetradecyl β-D-thiomaltoside; (Defaye, J. and Pederson, C., "Hydrogen Fluoride, Solvent and Reagent for Carbohydrate Conversion Technology," in Carbohydrates as Organic Raw Materials, 247-265 (F.W. Lichtenthaler, ed.), VCH Publishers, New York (1991); Ferenci, T.J. Bacteriol, 144:7-11 (1980)); alkyl thioglucosides such as 1-dodecyl or 1-octyl thio α- or β-D-glucopyranoside (Anatrace, Maumee, Ohio; see Saito, S. and Tsuchiya, T. Chem Pahrm. Bull. 33:503-508 (1985); alkyl thiosucrose (synthesized according to, e.g., Binder, TP and Robyt, JF Carbohydr. Res. 140:9-20 (1985)).Alkyl maltriosides (synthesized according to Koeltzow and Urfer); long-chain aliphatic carbonic acid amides of sucrose β-amino-alkyl ethers; (synthesized according to Austrian Patent No. 382,381 (1987); Chem. Abstr. 108:114719 (1988) and Gruber and Greber 95-116; derivatives of palatinose and isomaltamine linked to an alkyl chain by an amide bond (synthesized according to Kunz, M., "Hydrophilic Building Blocks Based on Sucrose as Intermediates for the Synthesis of Surfactants and Polymers" in Carbohydrates as Organic Raw Materials, pp. 127-153); derivatives of isomaltamine linked to an alkyl chain by a urea (synthesized according to Kunz); long-chain aliphatic ureidocarbonates of sucrose amino alkyl ethers (synthesized according to Gruber and Greber, pp. 95-116); and long-chain aliphatic amide carbonates of sucrose amino alkyl ethers (synthesized according to Austrian Patents 382, ​​381 (1987) Chem. Abstract 108:114719 (1988) and Gruber and Greber, pp. 95-116).

[0596] Some preferred glycosides include the sugars maltose, sucrose, glucose, and galactose linked by glycoside or ester linkages to alkyl chains of 6, 8, 10, 12, or 14 carbon atoms, e.g., hexyl-, octyl-, decyl-, dodecyl-, and tetradecyl-, maltoside, sucroside, glucoside, and galactosides. In the body, these glycosides are broken down into non-toxic alcohols or fatty acids and oligosaccharides or sugars. The above examples are illustrative of the types of alkyl glycosides that can be used in the methods claimed herein. However, the list is not intended to be exhaustive.

[0597] Generally, these surfactants (e.g., alkyl glycosides) are optionally designed to modulate bioavailability or are selected to modify the biological properties of the peptide, such as half-life, receptor selectivity, toxicity, biodistribution, solubility, stability (e.g., resistance to heat, hydrolysis, oxidation, and enzymatic degradation), purification and processing facilities, structural properties, spectroscopic properties, chemical and / or photochemical properties, catalytic ability, redox potential, ability to react with other molecules, e.g., covalently or non-covalently, etc.

[0598] <Surfactant> The term "surfactant" is derived from the contraction of "surface active agent." In pharmaceutical applications, surfactants are useful in liquid pharmaceutical formulations that serve many purposes because they act as dispersants, solubilizers, and wetting agents. Emulsifiers stabilize aqueous solutions of highly lipophilic or partially lipophilic substances. Solubilizers increase the solubility of components of a pharmaceutical composition, increasing the concentration that can be achieved. Wetting agents are additives that reduce the surface tension of a fluid, thus causing it to spread more easily on the surface to which it is applied, causing the surface to become more "wet" with the fluid. Wetting agents provide a means for the liquid formulation to achieve intimate contact with mucous membranes or other surfaces with which the pharmaceutical formulation comes into contact. Thus, surfactants may be useful additives for stabilizing formulations of the peptide products described herein as well as modifying the properties of the peptides themselves.

[0599] In specific embodiments, as described herein, similar synthetically accessible surfactants (e.g., alkyl glycosides dodecyl), tridecyl, and tetradecyl maltose glycosides or glucosides, alkyl glycosides, tridecanoates, and tetradecanoates are suitable for covalent attachment to peptides, as are the corresponding alkyl thioglycosides, which are synthetically available and stable enough for formulation development.

[0600] A wide range of physical 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 shown that, despite having the same length of hydrophobic tail in DM (Lopez, O. et al. (2002) Colloid Polym Sci 280:352 - 357), it is more than three times higher than that of DG. In this particular example, the identity of the polar region (disaccharide vs monosaccharide) affects surfactant behavior. In the case of surfactants linked to peptides, for example, the peptide products, as described herein, the peptide region can also contribute hydrophobic or hydrophilic characteristics to the overall molecule. Thus, tuning of physical surfactant properties may be used to achieve specific physical - pharmaceutical properties suitable for individual peptide targets.

[0601] <PEG modification> In some embodiments, the peptide products modified with the surfactants 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, the conjugation of large PEG chains prevents the filtration of peptides in the glomeruli in the kidney into the thin urine that results therefrom (Nestor, J.J. (Jr.) (2009) Current Medicinal Chemistry 16:4399 - 4418, Caliceti, P. and Veronese, F.M. (2003) Adv. Deliv Rev 55:1261 - 1277). In some embodiments, the optional PEG, hydrophilic chains enable the maintenance of the solubility and physical property balance of the peptide or protein, which is rendered hydrophobic by the attachment of the longer - term chains, the alkyl glycoside moiety.

[0602] PEGylation of proteins can also have potentially negative effects. Thus, PEGylation can cause a substantial loss of biological activity for some proteins. This may also be relevant for ligands for specific classes of receptors. In such cases, reversible PEGylation may be advantageous (Peleg-Shulman, T. et al. (2004) J Med Chem 47:4897-4904; Greenwald et al. (2003) Adv Drug Rev 55:217-250; Roberts, MJ and Harris, JM (1998) J Pharm Sci 87:1440-1445).

[0603] Additionally, increased molecular weight may prevent penetration of physiological barriers other than the glomerular membrane barrier. For example, it has been suggested that high molecular weight forms of PEGylation may prevent penetration into some tissues and, as a result, reduce therapeutic efficacy. Additionally, high molecular weight may prevent uptake across mucosal membrane barriers, including nasal, buccal, vaginal, oral, rectal, and pulmonary delivery. Even delayed uptake may be highly advantageous for stable administration of molecules to the lung, substantially extending the duration of action. The peptide and / or protein products described herein have increased oral mucosal bioavailability. This also allows longer-chain PEG modifications to be used in conjunction with surfactant modifications to achieve commercially significant bioavailability via intranasal or other oral mucosal routes.

[0604] In some embodiments, both long-chain and short-chain PEG polymers are suitable for modifying proteins and peptides as described herein. The administration of diabetes treatment by inhalation is a novel approach for drug delivery, as the lungs have a highly permeable barrier (e.g., Exubera). For this application, the preferred form of PEGylation, which slows lung barrier penetration, is in the lower molecular weight range of C10 to C400 (approximately 250 to 10,000 Da). Thus, while the primary route to elongation by PEG is achieving an "effective molecular weight" above the glomerular filtration cutoff (greater than 68 kDa), the use of shorter chains may be a route to extending lung retention for the treatment of pulmonary diseases and other respiratory conditions. Thus, PEG chains of approximately 500 to 3,000 Da are large enough to slow entry into the peripheral circulation, but insufficient to allow them to have a significantly extended circulation time. In some embodiments, PEGylation is applied to the covalently modified peptides and / or proteins described herein, which confer increased local efficacy in pulmonary tissue with the potential for reduced systemic side effects. In some of such embodiments, PEG chains in the range of about 750 to about 1500 da are collectively referred to as "PEG1K."

[0605] In addition, other polymers may be used in combination with the compounds described herein to optimize their physical properties. For example, poly(2-ethyl-2-oxazoline) conjugates have variable hydrophobicity and sufficient size to enhance duration of action (Mero, A. et al. (2008) J Control Release: 87-95). The attachment of such polymers to sugars results in a class of surfactants suitable for use in the modification of peptides and / or proteins described herein.

[0606] The polyethylene glycol chains are functionalized to allow their attachment to reactive groups on peptide and / or protein chains. Typical functional groups allow reaction with amino groups on the polyethylene glycol chain, carboxyl groups on the peptide with the corresponding carboxyl groups, or sulfhydryl groups, amino groups, or maleimide groups. In one embodiment, the PEG comprises a C10-C3000 chain. In another embodiment, the PEG has a molecular weight greater than 40,000 Daltons. In yet another embodiment, the PEG has a molecular weight less than 10,000 Daltons. PEG as a protein modification is well known in the art, and its use is described, for example, in U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192; and 4,179,337.

[0607] Nontraditional types of PEG chains are inherently amphiphilic; that is, they possess both hydrophilic and hydrophilic PEG structures but are modified to include hydrophobic regions, such as fatty acid esters and other hydrophobic moieties. See, for example, (Miller, MA et al. (2006) Bioconjug Chem 17:267-274); U.S. Patent No. 6,309,633 to Ekwuribe et al.; U.S. Patent No. 6,815,530 to Ekwuribe et al.; U.S. Patent No. 6,835,802 to Ekwuribe et al.). These amphiphilic PEG conjugates to proteins were originally developed to increase oral bioavailability, but they were relatively ineffective in this role. However, the use of such amphiphilic PEG conjugates with amphiphilic peptides confers significantly extended lung retention, extending the useful biological activity of these drugs. Preferred PEG chains are in the molecular weight range of 500 Da to 3000 Da. Detailed descriptions of methods for the synthesis of these conjugates are provided in the above-cited references. The full contents of which are incorporated herein by reference.

[0608] PEG entities lack functional groups that can be attached to target molecules, such as peptides. Therefore, to create PEGylated molecules, the PEG entity must first be functionalized, and the functionalized molecule is then used to attach the PEG entity to target molecules such as peptides (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) Drug Deliv Rev 54:459-476). In one embodiment, site-specific PEGylation can be achieved by the formation of a cysteine ​​surrogate on the peptide molecule. As described herein, target peptides can be synthesized by solid-phase methods, recombinant means, or other means.

[0609] Thus, in some embodiments, the peptide products described herein contain at least one cysteine ​​residue, a glycoside of an alkyl group on a cysteine ​​residue, a cysteine ​​residue, or other reactive amino acid residue, and a cysteine ​​residue or other reactive residue modified elsewhere in the molecule with specific PEGylation.

[0610] In another embodiment, Lys or other residues with nucleophilic side chains may be used for the attachment of PEG residues. This can be accomplished through the use of amide or carbamate linkages to the PEG carboxyl or PEG carbonate chains. See for example (Veronese, FM and Pasut, G. (2005) Drug Discovery Today 10:1451-1458). An alternative approach is to modify the Lys side chain, an amino function, by the attachment of an SH-containing residue, such as mercaptoacetyl, mercaptopropionyl, etc. (CO-CH-CH-CH-SH). Alternatively, the PEG chain can be incorporated at the C-terminus as an amide during synthesis. An additional method for attaching PEG chains utilizes reactions with His and Trp side chains. Other similar methods for modifying peptide chains to allow the attachment of PEG chains are known in the art and are embodied herein by (Roverts, MJ et al. (2002) Adv. Deliv. Rev 54:459-476).

[0611] <Prescription> In one embodiment, the covalently modified peptide or protein as provided herein is provided in a formulation that, for example, further reduces, prevents, or decreases aggregation of the peptide or protein association or composition, reduces peptide or protein self-association or self-association, or reduces association or aggregation with other peptides or proteins when processed in a subject.

[0612] Self-association at high protein concentrations is problematic in therapeutic formulations. For example, self-association increases the viscosity of concentrated monoclonal antibodies in aqueous solutions. Concentrated insulin preparations are inactivated by self-aggregation. These self-associating protein interactions, particularly at high protein concentrations, reduce, modulate, or eliminate the biological activity of many therapeutics (Clodfelter, DK et al. (1998) Genetically Engineered Bioproduction Facts 15:254-262). Therapeutic proteins formulated at high concentrations for delivery by infusion or other means can become physically unstable or insoluble as a result of these protein interactions.

[0613] A significant challenge in preparing peptide and protein formulations is developing manufacturable and stable dosage forms. Physical stability properties critical for processing and handling are often poorly characterized and difficult to predict. As determined by protein interaction and solubility properties, various physical instability phenomena, such as association, aggregation, crystallization, and precipitation, are encountered. This results in significant production, stability, analytical, and delivery challenges. The development of formulations for peptides and other protein drugs requiring high dosages (on the order of mg / kg) is necessary in many clinical situations. For example, using the SC route, approximately <1.5 mL is an acceptable administration volume. This may require protein concentrations of >100 mg / mL to achieve adequate dosage. Similar considerations exist when developing high-concentration lyophilized formulations for monoclonal antibodies. Generally, higher protein concentrations allow for smaller injection volumes to be used, which is crucial for patient comfort, convenience, and compliance. The surfactant-modified compounds described herein aim to minimize such aggregation events, which can be further facilitated through the use of small amounts of surfactants as described herein.

[0614] Because injection is an uncomfortable mode of administration for many people, other means of administering peptide therapeutics have been sought. For example, certain peptide and protein therapeutics can be administered by intranasal, buccal, oral, vaginal, inhalation, or other oral mucosal administration. Examples are nafarelin (Synarel®) and calcitonin, which are administered as commercial nasal spray formulations. Covalently modified peptides and / or proteins described herein are intended to facilitate such oral mucosal administration, and such formulations may be further facilitated through the use of small amounts of surfactants as described herein.

[0615] Typical formulation parameters include optimal hydrogen index, buffer, and stabilizing additive selection. Furthermore, lyophilized cake reconstitution is important for lyophilized or powder formulations. An even more important issue involves changes in the viscosity of protein formulations upon self-association. Viscosity changes can significantly modify delivery characteristics in spray (aerosol) delivery, for example, for intranasal spray, pulmonary spray, or oral spray. Furthermore, increased viscosity can make infusion delivery via syringe or IV line more difficult or impossible.

[0616] Numerous attempts to stabilize and maintain the integrity and biological activity of peptides have been reported. Some attempts have resulted in stabilization against thermal denaturation and aggregation, particularly for the insulin pump system. Polymeric surfactants have been described (Thurow, H. and Geisen, K. (1984) Diabetologia 27: 212-218; Chawla, A.S., et al. (1985) Diabetes 34: 420-424). The stabilization of insulin by these compounds was thought to be steric in nature. Other systems used include sugars (Arakawa, T. and Timasheff, SN (1982) Biochemistry 21: 6536-6544), osmolytes 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 modulating the hydrophobic interactions within protein or peptide molecules.

[0617] Various peptides, peptides, or proteins are described herein and can be modified with any of the covalent surfactant reagents described herein. Advantageously, the peptide modifications described herein include the covalent attachment of surfactants containing hydrophilic (e.g., sugars) and hydrophobic (e.g., alkyl chain) groups, thereby allowing for stabilization of the peptide at physiological conditions. In some embodiments, the covalent attachment of moieties containing hydrophilic and hydrophobic groups (e.g., glycoside surfactants) to the peptides and / or proteins described herein obviates the need to modify the amino acid sequence of the peptides and / or proteins to enhance stability (e.g., reduce aggregation).

[0618] In some embodiments, the formulations include at least one drug modified with a surfactant-derived agent described herein and further comprising a peptide associated with the surfactant in the formulation, where the surfactant is further comprised of, for example, a sugar, alkyl glycoside, or other excipient, and can be administered in a format selected from the group consisting of drops, sprays, aerosols, lyophilizates, spray dry products, injectables, and sustained-release formats. Sprays and aerosols can be achieved through the use of appropriate dispensers and can be administered via intranasal, transbuccal, inhalation, or other transmucosal routes. Lyophilizates can contain other compounds, such as mannitol, sugars, submicron anhydrous α-lactose, gelatin, biocompatible gels, or polymers. Sustained-release formats can be ocular inserts, erodible microparticles, hydrolyzable polymers, tumescent mucoadhesive microparticles, pH-sensitive microparticles, nanoparticle / latex systems, ion-exchange resins, and other polymer gels and implants (Ocusert, Alza Corp., California; Joshi, A., S. Ping and KJ Himmelstein, Patent Application WO 91 / 19481). Significant oral bioavailability can also be achieved.

[0619] The peptide and protein modifications described herein may mitigate, and in some cases, eliminate, the need for organic solvents. Trehalose, lactose, mannitol, and other sugars have been used to prevent aggregation. Aggregation of an anti-IgE humanized monoclonal antibody has been minimized by formulation with trehalose at molecular ratios ranging from 300:1 to 500:1 (excipient:protein) or higher. However, the resulting powders were either too cohesive 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 excipients discovered has limitations as a therapeutic additive, such as xenobiotic metabolism, irritation, or toxicity, or is costly. Contemplated for use with the covalently modified and / or proteins described herein are effective, non-irritating, and non-toxic excipients that do not require xenobiotic metabolism because they are themselves composed of natural sugars, fatty acids, or long-chain alcohols and can be used to minimize aggregation in aqueous solutions and in in situ protein formulations by aqueous reconstitution of dried peptides and / or physiological aqueous reconstitution with aqueous body fluids such as plasma or saliva.

[0620] Other formulation components may include buffers and physiological salts, non-toxic protease inhibitors such as aprotinin and soybean trypsin inhibitor, alpha-1-antitrypsin, and monoclonal antibodies that inactivate proteases. Buffers may also include organic substances such as acetate, citrate, gluconate, fumarate, malate, polylysine, polyglutamic acid, chitosan, dextran sulfate, and inorganic substances such as phosphate and sulfate. Such formulations may additionally contain low concentrations of bacteriostatic agents such as benzyl alcohol and the like.

[0621] Formulations suitable for intranasal administration also include solutions or suspensions of the modified peptides and / or proteins described herein in an acceptable evaporative solvent, such as a hydrofluoroalkane. Such formulations are suitable for administration from a metered dose inhaler (MDI) and have the advantages of lack of movement from the site of administration, low irritation, and no need for sterilization. Such formulations may also contain acceptable excipients or bulking agents, such as submicron anhydrous α-lactose.

[0622] In yet another aspect, the covalently modified and / or protein compositions described herein exhibit increased shelf life. As used herein, the phrase "shelf life" broadly describes the length of time a product can be stored before becoming unsuitable for use or consumption. The "shelf life" of a composition described herein can also refer to the time corresponding to an acceptable loss of quality of the composition. As used herein, compositional shelf life is distinguished from expiration date; "shelf life" relates to the quality of the compositions described herein, while "expiration date" is more concerned with the manufacturing and testing requirements of the composition. For example, a composition that has passed its "expiration date" may still be safe and effective, but optimal quality is no longer guaranteed by the manufacturer.

[0623] <Method> In one embodiment, a method is provided for administering an effective amount of a therapeutic composition described herein to a subject in need thereof. As used herein, the term "therapeutically effective amount" is interchangeable with the term "effective amount" for purposes of this specification and is determined by considerations known in the art. The amount must be effective to achieve the desired drug-mediated effect in the treated subject suffering from the disease. The therapeutically effective amount includes, but is not limited to, appropriate measurements selected by one of skill in the art, such as improved survival, more rapid recovery, symptom improvement or elimination, or other acceptable biomarkers or surrogate markers.

[0624] The compositions described herein are delivered to a vertebrate subject, such as, but not limited to, a human, in need of treatment. Moreover, depending on the condition being treated, these therapeutic compositions can be formulated and administered systemically or locally. Techniques for formulation and administration can be found in the latest edition of "Remington's Pharmaceutical Sciences" (Mack Publishing Co., Easton Pa.). Suitable routes can include parenteral delivery, including intramuscular, subcutaneous, intravenous, or intraperitoneal administration, as well as oral or transmucosal administration, such as intranasal, buccal, ocular, vaginal, or rectal administration.

[0625] In some embodiments of the method, the effective amount of the peptide product for administration is about 0.1 μg / kg / day to about 100.0 μg / kg / day, or 0.01 μg / kg / day to about 1 mg / kg / day, or 0.1 μg / kg / day to about 50 mg / kg / day. In some embodiments, the peptide product is administered parenterally. In some embodiments, the peptide product is administered subcutaneously. In some embodiments, the method of administration of the peptide product is intranasal inhalation.

[0626] It should be understood, however, that the specific dose level and frequency of administration for any particular subject in need of treatment may vary and will depend upon a variety of factors, such as the activity of the specific compound used, the metabolic stability and duration of action of that compound, age, weight, health, sex, diet, mode and time of administration, rate of excretion, drug combination, severity of the particular disease, and the host undergoing therapy.

[0627] In one embodiment, a method is provided for chemically modifying a molecule by covalent attachment to a surfactant to increase or maintain the biological activity of the composition or molecule, e.g., receptor binding or enzymatic activity. In some embodiments, the molecule is a peptide. The method can include further modifications, including covalent attachment of the molecule in the composition to a polymer, such as polyethylene glycol.

[0628] The methods include all aspects of the compositions described herein, including, but not limited to, compositions that reduce or eliminate the immunogenicity of peptide and / or protein drugs, are non-irritating, have antibacterial or antimicrobial activity, increase the stability or bioavailability of the drug, reduce the variability in bioavailability of the drug, prevent first-pass hepatic clearance, and reduce or eliminate any side effects. As used herein, the term "immunogenicity" refers to the ability of a particular substance or composition or drug to stimulate an immune response. The immunogenicity of the covalently modified and / or protein compounds described herein may be confirmed by methods known in the art.

[0629] Also provided are methods for administering a drug composition comprising a peptide covalently attached to at least one alkyl glycoside and delivered to a vertebrate, wherein the alkyl has 1 to 30 carbon atoms, or even is in the range of 6 to 16 carbon atoms, and the alkyl glycoside increases the stability, bioavailability, and / or duration of action of the drug.

[0630] In another embodiment, a method is provided for reducing or eliminating the immunogenicity of peptide and / or protein drugs by covalently attaching a peptide chain to at least one alkyl glycoside, wherein the alkyl has from 1 to 30 carbon atoms.

[0631] Throughout this application, various publications are referenced. Those skilled in the art will understand that the referenced disclosures of these publications are hereby incorporated by reference into this application.

[0632] <Treatment Method> In some embodiments, provided herein are methods for treating pain, including post-surgical or chronic pain, comprising administering a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula I, II, or III) to an individual in need thereof. In some such embodiments, the opioid analogs described herein are not addictive or habit-forming and / or are administered at lower doses than current agents (e.g., codeine) and have a longer duration of action than current agents.

[0633] In some embodiments, methods are provided for the prevention or treatment of diseases associated with hypoparathyroidism and / or decreased bone mass density, comprising administering to an individual in need thereof a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII). In some embodiments, diseases characterized by decreased bone mass density include, but are not limited to, osteoporosis, osteopenia, postmenopausal osteoporosis, Paget's disease, glucocorticoid-induced osteoporosis, senile osteoporosis, humoral hypercalcemia, or the like.

[0634] In some embodiments, provided herein are methods for treating hypoparathyroidism, comprising administering a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) to an individual in need thereof. In some embodiments, hypoparathyroidism is associated with decreased bone mass density.

[0635] Further provided herein are methods for stimulating bone repair and / or promoting implantation of bone implants, comprising administering to an individual in need thereof a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII).

[0636] In still further embodiments, provided herein are methods for increasing bone density and / or reducing the incidence of fractures (e.g., vertebral fractures, hip fractures, or the like), comprising administering to an individual in need thereof a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII).

[0637] In some embodiments, provided herein are methods for treating humoral hypercalcemia, comprising administering a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) to an individual in need thereof. In some embodiments, the humoral hypercalcemia is associated with a tumor. In some such embodiments, the peptide product (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) is an inverse agonist or antagonist of PTH or PTHrP.

[0638] In some embodiments of the above methods, the peptide and / or protein covalently attached to the surfactant is PTH, PTHrP, or an analog thereof. In some embodiments, the surfactant-modified peptide and / or protein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) is administered prophylactically to delay the onset of any disease associated with loss of bone density, including, but not limited to, osteoporosis, osteopenia, postmenopausal osteoporosis, Paget's disease, glucocorticoid-induced osteoporosis, senile osteoporosis, or the like. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) are administered therapeutically to slow the progression of any disease associated with loss of bone density, including, but not limited to, osteoporosis, osteopenia, postmenopausal osteoporosis, Paget's disease, glucocorticoid-induced osteoporosis, humoral hypercalcemia, or the like. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) are administered prophylactically and / or therapeutically to slow the progression of osteopenia to osteoporosis. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) are administered prophylactically and / or therapeutically to reduce or halt further loss of bone density, thereby stabilizing the disease.

[0639] In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) are administered parenterally. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) are administered subcutaneously. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) are administered by nasal insufflation.

[0640] In some embodiments of the above methods, the surfactant-modified peptide and / or protein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) has a longer duration of action than formulations containing currently known therapeutics (e.g., recombinant PTH, bisphosphonates, the antibody denosumab, or the like). In some embodiments, the surfactant-modified peptide and / or protein (e.g., a peptide product of Formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) is administered for a longer period (e.g., >2 years) than formulations containing currently known therapeutics (e.g., recombinant PTH, bisphosphonates, the antibody denosumab, or the like) while reducing or ameliorating side effects (e.g., intrajaw osteonecrosis, skin infections, or the like) associated with currently known therapeutics (e.g., recombinant PTH, bisphosphonates, the antibody denosumab, or the like). In some embodiments of the above methods, the surfactant-modified peptide and / or protein (e.g., a peptide product of formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) is an agonist of PTH or PTHrP. In some embodiments of the above methods, the surfactant-modified peptide and / or protein (e.g., a peptide product of formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) is an antagonist of PTH or PTHrP. In some embodiments of the above methods, the surfactant-modified peptide and / or protein (e.g., a peptide product of formula 2-IA, 2-III, 2-V, 2-VI, or 2-VII) is an inverse agonist of PTH or PTHrP.

[0641] In some embodiments, provided herein are methods for the prevention and / or treatment of diseases associated with decreased insulin sensitivity, comprising administering a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) to an individual in need thereof. In some embodiments, diseases characterized by decreased insulin sensitivity include, but are not limited to, metabolic syndrome, obesity-associated insulin resistance, hypertension, systemic inflammation associated with elevated C-reactive protein, diabetes, or the like.

[0642] Provided herein are methods for treating diseases associated with insulin resistance, including, but not limited to, obesity, metabolic syndrome, type 2 diabetes, hypertension, atherosclerosis, or the like, comprising administering a pharmaceutical composition comprising a peptide covalently attached to at least one alkyl glycoside and delivered to a vertebrate, wherein the alkyl has 1 to 30 carbon atoms, or even ranges from 6 to 18 carbon atoms (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V), wherein the covalent attachment of the alkyl glycoside to the peptide increases the stability, bioavailability, and / or duration of action of the drug.

[0643] Also described herein are methods for treating insulin resistance, comprising administering to an individual in need thereof a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V). In some embodiments, the insulin resistance is associated with metabolic syndrome (Syndrome X) and / or diabetes.

[0644] Further provided herein are methods for stimulating the body's resensitization to insulin, comprising administering to an individual in need thereof a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V).

[0645] In still further embodiments, provided herein are methods for increasing insulin sensitivity through weight loss, comprising administering to an individual in need thereof a therapeutically effective amount of a surfactant-modified peptide and / or protein product described herein (e.g., a peptide product of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V, and in Table 1 of Figures 1-1 through 1-7, and in Table 2 of Figures 2-1 through 2-4).

[0646] Also provided herein are methods for treating diabetes or prediabetes, comprising administering to an individual in need thereof a therapeutically effective amount of a peptide product described above and herein, and in Table 1 of Figures 1-1 to 1-7 and Table 2 of Figures 2-1 to 2-4.

[0647] Provided herein are methods for treating or delaying the progression or onset of a disease selected from diabetes, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, insulin resistance, hyperglycemia, hyperinsulinemia, metabolic syndrome, diabetic complications, high blood levels of free fatty acids or glycerol, hyperlipidemia, obesity, hypertriglyceridemia, atherosclerosis, acute cardiovascular syndrome, infarction, and ischemia-reperfusion hypertension, comprising administering to an individual in need thereof a therapeutically effective amount of a peptide product described herein and in Table 1 of Figures 1-1 to 1-7 and in Table 2 of Figures 2-1 to 2-4. In additional embodiments, provided are methods for treating delayed wound healing, comprising administering to an individual in need thereof a therapeutically effective amount of a peptide product described herein and in Table 1 of Figures 1-1 to 1-7 and in Table 2 of Figures 2-1 to 2-4.

[0648] In one embodiment, the disease to be treated is diabetes. In one embodiment, the disease to be treated is insulin resistance. In one embodiment, the disease to be treated is metabolic syndrome. In one embodiment, the effective amount of the peptide is from about 0.1 μg / kg / day to about 100.0 μg / kg / day.

[0649] 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 intranasal inhalation.

[0650] Further provided herein are methods for reducing weight gain and inducing weight loss, comprising administering to an individual in need thereof a therapeutically effective amount of a peptide product described herein and in Table 1 of Figures 1-1 to 1-7 and Table 2 of Figures 2-1 to 2-4. In some embodiments, the weight gain is associated with metabolic syndrome.

[0651] Further provided herein is a method for treating hypoglycemia, comprising administering to an individual in need thereof a therapeutically effective amount of a peptide product described herein and in Table 1 of Figures 1-1 to 1-7 and Table 2 of Figures 2-1 to 2-4.

[0652] A method for treating diabetes is also provided herein, comprising administering to an individual in need thereof a therapeutically effective amount of a peptide product described herein and in Table 1 of Figures 1-1 to 1-7 and Table 2 of Figures 2-1 to 2-4, wherein the therapeutic agent is selected from a diabetes agent, an anti-obesity agent, a satiety agent, an anti-inflammatory agent, an anti-hypertensive agent, an anti-atherosclerotic agent, and a lipid-lowering agent.

[0653] In some embodiments of the above methods, the peptide and / or protein covalently attached to the surfactant is a glucagon or GLP-1 peptide, or analog thereof. In some embodiments, the surfactant-modified peptide and / or protein (e.g., a peptide product of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) is administered prophylactically to delay the onset of any disease associated with insulin resistance, including, but not limited to, metabolic syndrome, hypertension, diabetes, type 2 diabetes, gestational diabetes, hyperlipidemia, atherosclerosis, systemic inflammation, or the like. In some embodiments, the surfactant-modified peptide and / or protein (e.g., a peptide product of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) is administered therapeutically to delay the progression of any disease associated with metabolic syndrome, hypertension, diabetes, type 2 diabetes, gestational diabetes, hyperlipidemia, atherosclerosis, systemic inflammation, or the like. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) are administered prophylactically and / or therapeutically to slow the progression of insulin resistance to diabetes. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) are administered prophylactically and / or therapeutically to reduce or halt further loss of insulin resistance, thereby stabilizing the disease.

[0654] In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) are administered parenterally. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) are administered subcutaneously. In some embodiments, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or Formula 3-V) are administered by nasal insufflation.

[0655] In some embodiments of the above methods, the surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-I-A, 3-III-A, 3-III-B or Formula 3-V) have a longer duration of action compared to agents including currently known therapeutics (e.g., exenatide, metformin, or the like).

[0656] <Combination Therapy with PTH Analogs> In some embodiments of the above methods, the surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-I-A, 2-III, 2-V, 2-VI or 2-VII) are administered in combination with an antiresorptive agent including a bisphosphonate (e.g., alendronate) or a strontium salt, or a substance having an estrogen-like effect (e.g., estrogen), or a selective estrogen receptor modulator (e.g., raloxifene, tamoxifen, droloxifene, toremifene, idoxifene, or levormeloxifene), or a calcitonin-like substance (e.g., calcitonin), or a vitamin D analog, or a calcium salt. The therapeutic agents are optionally administered simultaneously or sequentially in any order. As an example, in some embodiments of the above methods, a first regimen of the surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-I-A, 2-III, 2-V, 2-VI or 2-VII) is administered to an individual who needs them, followed by a second regimen of bisphosphonate therapy. As an example, in some other embodiments, a first regimen of the surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 2-I-A, 2-III, 2-V, 2-VI or 2-VII) is administered to an individual who needs them, followed by a drug-free period, and then followed by a second regimen of an estrogen receptor modulator.

[0657] <Combination Therapy with GLP Analogs> In some embodiments of the above methods, the surfactant-modified peptide and / or protein (e.g., a peptide product of Formula 3-IA, 3-III-A, 3-III-B, or 3-V) is administered in combination with other methods of treating metabolic syndrome selected from the group consisting of antidiabetic agents, antiobesity agents, antihypertensive agents, antiatherosclerotic agents, and lipid-lowering agents. By way of example, effective antidiabetic agents suitable for administration in combination with the surfactant-modified peptide and / or protein products described herein include biguanides, sulfonylurea derivatives, glucosidase inhibitors, PPARγ agonists, PPARα / γ dual agonists, aP2 inhibitors, DPP4 inhibitors, insulin sensitizers, GLP-1 analogs, insulin, and meglitinides. Additional examples include metformin, glyburide, glimepiride, glipizide, chlorpropamide, gliclazide, acarbose, miglitol, pioglitazone, troglitazone, rosiglitazone, muraglitazar, insulin, GI-262570, isaglitazone, JTT-501, NN-2344, L895 645, YM-440, R-119702, A19677, repaglinide, nateglinide, KAD 1129, AR-HO 39242, GW-40 I 5 44, KRP2 I 7, AC2993, LY3 I 5902, NVP-DPP-728A, and saxagliptin.

[0658] In some embodiments of the above methods, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or 3-V) are administered in combination with other methods of treating metabolic syndrome selected from the group of effective anti-obesity agents. By way of example, 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 these classes include orlistat, AfL-962, A19671, L750355, CP331648, sibutramine, topiramate, axokine, dexamphetamine, phentermine, phenylpropanolamine, rimonabant (SR1 4I7164) and mazindol.

[0659] In some embodiments of the above methods, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or 3-V) are administered in combination with other therapies for metabolic syndrome selected from the group of effective lipid-lowering agents. By way of example, effective lipid-lowering 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 synthase inhibitors, fibric acid derivatives, upregulators of LDL receptor activity, lipoxygenase inhibitors, and ACAT inhibitors. Specific examples from these classes include pravastatin, lovastatin, simvastatin, atorvastatin, cerivastatin, fluvastatin, nisvastatin, visastatin, fenofibrate, gemfibrozil, clofibrate, avasimibe, TS-962, MD-700, CP-52941 4 and LY295 427.

[0660] In some embodiments of the above methods, surfactant-modified peptides and / or proteins (e.g., peptide products of Formula 3-IA, 3-III-A, 3-III-B, or 3-V) are administered in combination with peptide hormones and analogs thereof known to induce prosatiety in animal models and humans. Combinations of the peptide products described herein with long-acting satiety agents for the treatment of obesity are contemplated within the scope of the embodiments presented herein. Examples of such peptide satiety agents include GLP-1, pancreatic polypeptide (PP), cholecystokinin (CCK), peptide YY (PYY), amylin, calcitonin, OXM, neuropeptide Y (NPY), and analogs thereof (Bloom, SR, et al. (2008) Mol Interv 8: 82-98; Field, BC, et al. (2009) Br J Clin Pharmacol 68: 830-843).

[0661] Methods for the treatment of obesity comprising the administration of the peptide products described herein in combination with peptide hormones, including but not limited to, leptin, ghrelin, and CART (cocaine- and amphetamine-regulated transcription) analogs and antagonists, are also contemplated within the scope of the embodiments presented herein.

[0662] Additional peptide products in the body are associated with adipocytes or obese states (adipokines) and are known to have pro-inflammatory effects (Gonzalez-Periz, A. and Claria, J. (2010) Scientific World Journal 10: 832-856). Such agents have additional beneficial effects when used in combination with the peptide products described herein. Examples of agents that provide beneficial effects when used in combination with the peptide products described herein include analogs and antagonists of adiponectin, chemerin, visfatin, nesfatin, omentin, resistin, TNF-alpha, IL-6, and obestatin.

[0663] <Medication> The covalently modified and / or protein compounds described herein can be administered in any amount to provide beneficial therapeutic effects in many disease states. In some embodiments, the covalently modified and / or protein compounds described herein are useful in treating inflammation. In embodiments, the compounds provided herein provide beneficial activity in modulating post-surgical or chronic pain. In embodiments, the peptides of the present invention are administered to patients at concentrations higher or lower than other forms of treatment to modulate pain. In yet other embodiments, the peptides of the present invention are administered with other compounds to provide a synergistic therapeutic effect.

[0664] Representative delivery regimens include oral, parenteral (including subcutaneous, intramuscular, and intravenous injection), rectal, buccal (including sublingual), transdermal, inhalation, ocular, and intranasal. An attractive and widely used method for delivery of pe...

Claims

1. A peptide product comprising a surfactant X covalently attached to a peptide, the peptide comprising a linker amino acid U and at least one other amino acid: 【Chemistry 1】 In the formula, X is: 【Chemistry 2】 and During the ceremony, A is a hydrophobic group; and A peptide product, characterized in that B is a hydrophilic group covalently attached to the peptide via a linker amino acid U.

2. 2. The peptide product according to claim 1, characterized in that the peptide product is synthesized by reaction of a functionalized surfactant with a peptide.

3. A is a substituted or unsubstituted C １ -C ３０ 2. The peptide product of claim 1, which is characterized in that it is a moiety containing an alkyl chain, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a steroid nucleus.

4. 2. The peptide product according to claim 1, characterized in that B is a polyol.

5. 5. The peptide product according to claim 4, wherein the polyol is a sugar.

6. 6. The peptide product according to claim 5, characterized in that the sugars are selected from glucose, mannose, maltose, glucuronic acid, galacturonic acid, diglucuronic acid and malturonic acid.

7. 2. A peptide product according to claim 1, characterized in that the surfactant is a surfactant of the 1-alkylglycoside class.

8. 8. A peptide product according to any one of claims 1 to 7, characterized in that the hydrophilic group in the surfactant is attached to the peptide via an amide bond.

9. The surfactant X may be 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-hexadecyl beta-D-diglucuronic acid, 1-tetradecyl beta-D-diglucuronic acid, 1-dodecyl beta-D-diglucuronic acid, 1-decyl beta-D-diglucuronic acid, 1-octyl beta-D-diglucuronic acid or functionalized 1-eicosyl beta-D-glucuronic acid, 1-octadecyl beta-D-glucuronic acid, 1-hexadecyl beta-D-diglucuronic acid, 1-tetradecyl beta-D-diglucuronic acid, 1-dodecyl beta-D-diglucuronic acid, 1-decyl beta-D-diglucuronic acid, 1-octyl beta-D-diglucuronic acid or functionalized 1-eicosyl beta-D-glucose, 1-octadecyl beta-D-glucuronic acid, 1- 8. Peptide product according to any one of claims 1 to 7, characterized in that it consists of hexadecyl beta-D-glucose, 1-tetradecyl beta-D-glucose, 1-dodecyl beta-D-glucose, 1-decyl beta-D-glucose, 1-octyl beta-D-glucose, 1-eicosyl beta-D-maltoside, 1-octadecyl beta-D-maltoside, 1-hexadecyl beta-D-maltoside, 1-dodecyl beta-D-maltoside, 1-decyl beta-D-maltoside, 1-octyl beta-D-maltoside etc.

10. A peptide product according to any one of claims 1 to 7, characterized in that the peptide is an opioid peptide.

11. The peptide product has the structure of Formula IA: 【Chemistry 3】 During the ceremony: aa １ , aa ２ , aa ３ , aa ４ , and aa ５ each independently represents a non-existent, D or L natural or unnatural amino acid, an N-alkylated amino acid, an N-acetylated amino acid, a CαR ３ an amino acid, a Ψ amino acid, or a linker amino acid U covalently attached to a surfactant X; Z is -OH, -NH ２ Or -NHR ３ and Each R ３ is independently a substituted or unsubstituted C １ -C １２ a branched or linear alkyl, a PEG chain less than 10 Da, or a substituted or unsubstituted aralkyl chain; However, aa １ , aa ２ , aa ３ , aa ４ , and aa ５ is a linker amino acid U covalently attached to surfactant X; and further, aa １ , aa ２ , aa ３ , aa ４ , and aa ５ 11. The peptide product according to claim 10, characterized in that all of the following are not absent:

12. A peptide product having the structure of Formula II: 【Chemistry 4】 During the ceremony: aa １ , aa ２ , aa ３ , aa ４ , and aa ５ each independently represents a non-existent, D or L natural or unnatural amino acid, an N-alkylated amino acid, an N-acetylated amino acid, a CαR ３ an amino acid, a Ψ amino acid, or a linker amino acid U covalently attached to a surfactant X; Z is -OH, -NH ２ Or -NHR ３ and Each R ３ is independently a substituted or unsubstituted C １ -C １２ a branched or linear alkyl, a PEG chain of less than 10 Da, or a substituted or unsubstituted aralkyl chain; and X is 【Chemistry 5】 and During the ceremony, A is a hydrophobic group; and B is a hydrophilic group covalently attached to the peptide via the linker amino acid U; However, aa １ , aa ２ , aa ３ , aa ４ , and aa ５ is a linker amino acid U covalently attached to surfactant X; and further, aa １ , aa ２ , aa ３ , aa ４ , and aa ５ provided that not all of the following are absent.

13. The peptide product has the structure of Formula III: 【Chemistry 6】 During the ceremony: aa １ is Tyr, Dmt, N-R ３ -Tyr, N-R ３ -Dmt,N-(R ３ ) ２ -Tyr, or N-(R ３ ) ２ - Dmt; aa ２ is Pro, D-Arg, DU(X), D-Ala, Tic, or Tic(Ψ[CH2-NH]); aa ３ is Phe, Trp, Tmp, D- or L-Nal(1), D- or L-Nal(2), CαMePhe, or Ψ-Phe; aa ４ is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), U(X), D- or L-CαMeU(X); aa ５ is absent or is Pro, Aib, U(X), D- or L-CαMeU(X); and U is a dibasic natural or unnatural amino acid, a thiol-containing natural or unnatural amino acid, -N ３ unnatural amino acids containing a group, unnatural amino acids containing an acetylene group, or -NH-C(=O)-CH ２ -Br or -(CH ２ ) ｍ 13. The peptide product according to claim 12, characterized in that m is an unnatural amino acid containing maleimide, where m is 1-10.

14. X has the structure: 【Chemistry 7】 During the ceremony: A is a substituted or unsubstituted C １ -C ３０ an alkyl chain, a substituted or unsubstituted alkoxyaryl group, a substituted or unsubstituted aralkyl group, or a moiety containing a steroid nucleus; R １ｂ , R １ｃ , and R １ｄ each independently at each occurrence is H, a protecting group, or a substituted or unsubstituted C １ -C ３０ is an alkyl group; 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-; W ２ is -O-, or -S-; R ２ is a single bond, C ２ -C ４ Alkenes, C ２ -C ４ Alkyne, or -(CH ２ ) m-maleimide; and 14. The peptide product according to claim 13, characterized in that m is 1-10.

15. 14. The peptide product of claim 13, wherein X has the following structure: 【Chemistry 8】

16. 14. The peptide product of claim 13, wherein X has the following structure: 【Chemistry 9】

17. X has the structure: 【Chemistry 10】 During the ceremony: A is a substituted or unsubstituted C １ -C ３０ an alkyl group, or a moiety containing a steroid nucleus; R １ｂ , R １ｃ , and R １ｄ each independently at each occurrence is H, a protecting group, or a substituted or unsubstituted C １ -C ３０ is an alkyl group; W １ is -(C=O)-NH-; W ２ is -O-; R ２ The peptide product according to claim 13, characterized in that: is a single bond.

18. X has the structure: 【Chemistry 11】 During the ceremony: A is a substituted or unsubstituted C １ -C ３０ 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 The peptide product according to claim 13, characterized in that: is a single bond.

19. A is a substituted or unsubstituted C １ -C ２０ 19. A peptide product according to any one of claims 14 to 18, characterized in that it is an alkyl group.

20. The product (SEQ.ID.NO.149): During the ceremony: U is a dibasic natural or unnatural amino acid; X is a surfactant of the 1-alkyl glycoside class, where the 1-alkyl is a substituted or unsubstituted C １ -C ２０ is an alkyl or substituted or unsubstituted alkoxyaryl substituent; Z is NH ２ and aa １ is Tyr, Dmt, Nα-Me-Tyr, Nα-Me-Tyr, N,Nα-diMe-Tyr, or N,Nα-diMe-Dmt; aa ２ is Pro, D-Arg, DU(X), D-Ala, Tic, or Tic(Ψ[CH2-NH]); aa ３ is Phe, Trp, Tmp, D- or L-Nal(1), D- or L-Nal(2), CαMePhe, or Ψ-Phe; aa ４ is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), U(X), D- or L-CαMeU(X); aa ５ The peptide product according to claim 13, characterized in that: is absent or is Pro, Aib, U(X), D- or L-CαMeU(X).

21. The product (SEQ.ID.NO.150): During the ceremony: X consists of 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid; Z is NH ２ and aa １ is Dmt; aa ２ is Pro, D-Lys(X), Tic, or Tic(Ψ[CH2-NH]); aa ３ is Phe, Tmp, D- or L-Nal (1), D- or L-Nal (2), or Ψ-Phe; aa ４ is Phe, D- or L-Nal (1), D- or L-Nal (2), or Lys (X); aa ５ The peptide product according to claim 20, characterized in that: is absent or is Pro, Aib, Lys(X), D- or L-CαMeLys(X).

22. The product (SEQ.ID.NO.151), During the ceremony: aa １ is Dmt; aa ２ is Pro; aa ３ is Phe, or Tmp; aa ４ is Phe, or Lys(X); aa ５ The peptide product according to claim 21, characterized in that: is absent or is Aib, Lys(X), D- or L-CαMeLys(X).

23. The product (SEQ.ID.NO.152): During the ceremony: aa １ is Dmt; aa ２ is Pro; aa ３ is Phe, or Tmp; aa ４ is Phe, or Lys(X); aa ５ The peptide product according to claim 22, characterized in that: is absent or is Lys(X).

24. The product (SEQ.ID.NO.153): During the ceremony: U is a dibasic natural or unnatural amino acid; X is a surfactant of the 1-alkylglycoside class, where the 1-alkyl group of the 1-alkylglycoside is a substituted or unsubstituted C １ -C ２０ is an alkyl or substituted or unsubstituted alkoxyaryl substituent; aa １ is Tyr, Dmt, N-R ３ -Tyr, N-R ３ -Tyr, N-(R ３ ) ２ -Tyr, or N-(R ３ ) ２ - Dmt; aa ２ is D-Arg, or D-U(X); aa ３ is Phe, D- or L-Nal(1), D- or L-Nal(2), or Tmp; aa ４ is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), U(X), D- or L-CαMeU(X); aa ５ The peptide product according to claim 13, characterized in that: is absent, Pro, or U(X).

25. The product (SEQ.ID.NO.154): During the ceremony: X is a surfactant of the 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid class; aa １ is Dmt, Nα-Me-Dmt, or N,Nα-Me-Dmt; aa ２ is D-Arg, D-Lys(X), or D-Orn(X); aa ３ is Phe, or Tmp; aa ４ is Phe, Tmp, Lys(X), or Orn(X); aa ５ 25. The peptide product according to claim 24, characterized in that: is absent or is Pro, Lys(X), or Orn(X).

26. The product (SEQ.ID.NO.155): During the ceremony: U is a dibasic natural or unnatural amino acid; X is a surfactant of the 1-alkyl glycoside class, where the 1-alkyl is a substituted or unsubstituted C １ -C ２０ is an alkyl or substituted or unsubstituted alkoxyaryloxy substituent; Z is NH ２ and aa １ is Tyr, Dmt, N-R ３ -Tyr, N-R ３ -Dmt,N-(R ３ ) ２ -Tyr, or N-(R ３ ) ２ - Dmt; aa ２ is Tic, or Tic(Ψ[CH2-NH]); aa ３ , aa ２ When Tic(Ψ[CH2-NH]), then Ψ-Phe; aa ４ is Phe, Tmp, D- or L-Nal (1), D- or L-Nal (2), or U (X); aa ５ 26. The peptide product according to claim 24 or 25, characterized in that: is absent, Pro, Aib, or U(X).

27. The product (SEQ.ID.NO.156): During the ceremony: X is a surfactant of the 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid class; aa １ is Tyr, Dmt, Nα-Me-Tyr, Nα-alkyl-Dmt, N,Nα-diMe-Tyr, or N,Nα-Me-Dmt; aa ２ is Tic, or Tic(Ψ[CH2-NH]); aa ３ , aa ２ is Tic(Ψ[CH2-NH]), Phe, or TMP, then Ψ-Phe; aa ４ is Phe, Tmp, D- or L-Nal(1), D- or L-Nal(2), Lys(X), or Orn(X); aa ５ 27. The peptide product according to claim 26, characterized in that: is absent or is Aib, Lys(X), or Orn(X).

28. The product (SEQ.ID.NO.157): During the ceremony: X consists of 1-alkyl glucuronic acid or 1-alkyl diglucuronic acid; aa ２ is Tic, or Tic(Ψ[CH2-NH]); aa ３ is Phe or Ψ-Phe; aa ４ is Lys(X); aa ５ The peptide product according to claim 13, characterized in that it is absent.

29. The product (SEQ.ID.NO.158): During the ceremony: X consists of 1-alkyl glucuronic acid; aa ２ is Tic: aa ３ is Phe; aa ４ is Lys(X); aa ５ 29. The peptide product according to claim 28, characterized in that it is absent.

30. The product (SEQ.ID.NO.159): During the ceremony: X consists of a 1-alkyl glucuronic acid selected from 1-methyl beta-D-glucuronic acid, 1-octyl beta-D-glucuronic acid, 1-dodecyl beta-D-glucuronic acid, 1-tetradecyl beta-D-glucuronic acid, 1-hexadecyl beta-D-glucuronic acid, and 1-octadecyl beta-D-glucuronic acid; aa ２ is Tic; aa ３ is Phe; aa ４ is Lys(X); aa ５ 30. The peptide product according to claim 29, characterized in that it is absent.

31. H-Dmt-Tic-Phe-Lys(N-epsilon-1-methyl beta-D-glucuronyl)-NH ２ 31. The peptide product according to claim 30, characterized in that it is (SEQ.ID.NO.78).

32. H-Dmt-Tic-Phe-Lys(N-epsilon-1-octyl beta-D-glucuronyl)-NH ２ 31. The peptide product according to claim 30, characterized in that it is (SEQ.ID.NO.80).

33. H-Dmt-Tic-Phe-Lys(N-epsilon-1-dodecyl beta-D-glucuronyl)-NH ２ 31. The peptide product according to claim 30, characterized in that it is (SEQ.ID.NO.79).

34. H-Dmt-Tic-Phe-Lys(N-epsilon-1-tetradecyl beta-D-glucuronyl)-NH ２ 31. The peptide product according to claim 30, characterized in that it is (SEQ.ID.NO.160).

35. H-Dmt-Tic-Phe-Lys(N-epsilon-1-hexadecyl beta-D-glucuronyl)-NH ２ 31. The peptide product according to claim 30, characterized in that it is (SEQ.ID.NO.82).

36. H-Dmt-Tic-Phe-Lys(N-epsilon-1-octadecyl beta-D-glucuronyl)-NH ２ 31. The peptide product according to claim 30, characterized in that it is (SEQ.ID.NO.83).

37. A compound selected from the compounds of Table 1 in FIG.

38. 38. A pharmaceutical composition comprising a therapeutically effective amount of the peptide product of any one of claims 1 to 37, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient.

39. 1. A method for improving the pharmaceutical and therapeutic response of a peptide, thereby improving its duration of action, its bioavailability, or its stability in formulations, characterized in that the method comprises the step of covalently attaching to the peptide a surfactant X, wherein X is as defined in claim 1.

40. 40. A method of treating pain comprising administration of a therapeutically effective amount of a peptide product according to any one of claims 1 to 37.

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